Publications by authors named "Huimin Yu"

196 Publications

Evaluation of the relationship among dental fear, scaling and root planing and periodontal status using periodontitis stages: A retrospective study.

J Dent Sci 2022 Jan 21;17(1):293-299. Epub 2021 Apr 21.

Department of Periodontology, Stomatological Hospital, Southern Medical University, Guangzhou, China.

Background/purpose: Patients with periodontal disease have higher dental fear levels, which may have negative effects on their clinical outcome during scaling and root planing (SRP). The present study used the new classification of periodontitis and validated questionnaires to assess the relationship among dental fear, SRP pain and periodontal status.

Materials And Methods: A total of 120 periodontitis patients were enrolled and staging according to the new classification of periodontitis. SRP was performed, and the visual analog scale (VAS) to assess pain was used with every patient after treatment. Questionnaires, including Corah's Dental Anxiety Scale (DAS), Dental Fear Survey (DFS), and short-form Dental Anxiety Inventory (S-DAI) were implemented from the first attendance and subsequent visits after 6 months. The patients were grouped by DAS scores. The statistical analysis was performed using T-test, chi-square, Pearson and Spearman correlative analysis.

Results: Compared to pre-SRP treatment, the dental fear level on DFS was decreased in the posttreatment period for all periodontitis stages. There were no statistically significant differences in S-DAI and DAS between pretreatment and posttreatment periods in stage I and II; meanwhile, there were statistically differences in stage III and IV. The correlation among periodontitis stages, VAS and dental fear level was significant. The proportion of high periodontitis stages was increased in high dental fear group.

Conclusion: SRP can reduce dental fear levels in all periodontitis stages, especially in stage III and IV. Correlations exist among periodontal status, dental fear and SRP pain. High dental fear is associated with poor periodontal status.
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http://dx.doi.org/10.1016/j.jds.2021.04.002DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8739248PMC
January 2022

Therapeutic carrier based on solanesol and hyaluronate for synergistic tumor treatment.

Int J Biol Macromol 2022 Jan 5;201:20-28. Epub 2022 Jan 5.

Institute of Pharmacy, School of Pharmacy, Henan University, Kaifeng 475004, China. Electronic address:

The administration of nanodrugs can lead to metabolism related systemic toxicity due to the use of inert carriers in large quantities. Carrier materials that offer therapeutic effects are therefore a promising means of addressing this limitation. Herein, a hyaluronate-based nanocarrier was prepared from hyaluronic acid (HA) and solanesol. Solanesyl thiosalicylate (STS) derived from solanesol has certain antitumor effects and was used to modify HA. The conjugate (HA-STS) self-assembled into nanoparticles acting as a drug carrier. The synthesis of the conjugates was confirmed by H NMR spectroscopy. Doxorubicin (DOX) was loaded into the HA-STS nanoparticles with a relatively high content of 6.0%. pH-sensitive drug release behavior was achieved by introducing a hydroazone bond between STS and HA. A cytotoxicity assay indicated that the blank nanoparticles had an antitumor effect, which was enhanced by loading with an additional drug. Moreover, in vivo antitumor experiments indicated that the HA-STS-DOX showed superior tumor inhibition compared with free DOX, as well as lower cardiotoxicity and hepatotoxicity, demonstrating the advantages of the bioactive drug vehicles in cancer therapy.
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http://dx.doi.org/10.1016/j.ijbiomac.2021.12.194DOI Listing
January 2022

Indirect Pathway Metabolic Engineering Strategies for Enhanced Biosynthesis of Hyaluronic Acid in Engineered .

Front Bioeng Biotechnol 2021 20;9:768490. Epub 2021 Dec 20.

Key Laboratory for Industrial Biocatalysis of the Ministry of Education, Department of Chemical Engineering, Tsinghua University, Beijing, China.

Hyaluronic acid (HA) is composed of alternating d-glucuronic acid and -acetyl-d-glucosamine, with excellent biocompatibility and water retention capacity. To achieve heterologous biosynthesis of HA, , a safe GRAS (generally recognized as safe) host, was utilized and metabolically engineered previously. In this work, to achieve further enhancement of HA yield, four strategies were proposed and performed separately first, i.e., (1) improvement of glucose uptake via gene knockout, releasing the inhibition of transporter IolT1/IolT2 and glucokinases; (2) intensification of cardiolipin synthesis through overexpression of genes // involved in cardiolipin synthesis; (3) duly expressed hemoglobin in genome, enhancing HA titer coupled with more ATP and improved NAD/NADH (>7.5) ratio; and (4) identification of the importance of glutamine for HA synthesis through transcriptome analyses and then enhancement of the HA titer via its supplement. After that, we combined different strategies together to further increase the HA titer. As a result, one of the optimal recombinant strains, Cg-dR-CLS, yielded 32 g/L of HA at 60 h in a fed-batch culture, which was increased by 30% compared with that of the starting strain. This high value of HA titer will enable the industrial production of HA via the engineered .
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http://dx.doi.org/10.3389/fbioe.2021.768490DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8721151PMC
December 2021

3D-Printed Template-Guided I Seed Brachytherapy: A Salvage Approach for Locoregional Refractory Recurrence of Papillary Thyroid Cancer.

Eur Thyroid J 2021 Nov 4;10(6):504-510. Epub 2021 Oct 4.

Department of Radiology, The 4th Affiliated Hospital of Hebei Medical University, Shijiazhuang, China.

Background: Papillary thyroid carcinoma (PTC) is typically treated with surgical resection, even in recurrent cases. However, some cases of recurrent PTC are refractory to the conventionally used locoregional radiotherapy and resection methods. I seed permanent brachytherapy has emerged as a promising alternative for such PTCs, but no effective brachytherapy protocol has been reported for tumors with a huge volume, liquefaction, necrosis, and skin invasion.

Case Presentation: A 47-year-old man presented with recurrence 8 years after 2 thyroidectomy procedures for PTC and recurrent PTC. The tumor measured 6 × 7 × 8 cm and exhibited liquefaction, necrosis, and skin invasion. The patient was treated at our hospital from December 2017 to November 2018. He received one round of I seed temporary brachytherapy and 4 rounds of I seed permanent implantation. The activity of the seeds was 0.3-3.0 mCi, and the total dose delivered to the tumor was 145 Gy. The recurrent tumor was successfully removed by I seed brachytherapy guided with a 3D-printed template and ultrasound and CT scanning. The refractory tumor healed uneventfully after I seed brachytherapy without recurrence over the 25-month follow-up.

Conclusions: To the best of our knowledge, this is the first reported case of a large thyroid carcinoma that was effectively treated by 3D-printed template-guided I seed brachytherapy.
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http://dx.doi.org/10.1159/000519572DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8647112PMC
November 2021

Zinc uptake and replenishment mechanisms during repeated phytoextraction using Sedum plumbizincicola revealed by stable isotope fractionation.

Sci Total Environ 2022 Feb 29;806(Pt 3):151306. Epub 2021 Oct 29.

CAS Key Laboratory of Soil Environment and Pollution Remediation, Institute of Soil Science, Chinese Academy of Sciences, Nanjing 210008, China.

Improving phytoremediation techniques requires a thorough understanding of the mechanisms of plant uptake and the replenishment of the bioavailable pool of the target element, and this may be effectively explored using stable isotope methods. A repeated phytoextraction experiment over five successive crops of cadmium (Cd) and zinc (Zn) hyperaccumulator Sedum plumbizincicola X.H. Guo et S.B. Zhou ex L.H. Wu (Crassulaceae) was conducted using four agricultural soils differing in soil pH and clay content. The isotopic composition of total Zn and NHOAc-extractable Zn in soils before phytoextraction and after the fifth crop were determined, together with Zn in shoot samples in the first crop. S. plumbizincicola preferentially took up light Zn isotopes from the NHOAc-extractable pool (ΔZn = -0.42 to -0.16‰), indicating the predominance of Zn low-affinity transport. However, after long-term phytoextraction NHOAc-extractable Zn became isotopically lighter than prior to phytoextraction in three of the soils (ΔZn = -0.39 to -0.10‰). This was resulted from the equilibrium replenishment of Zn bound to iron (Fe) and manganese (Mn) oxides based on Zn isotopic and chemical speciation analysis. Zinc showed opposite fractionation patterns to Cd in the same plant-soil system with heavy Cd isotope enrichment in S. plumbizincicola (ΔCd = 0.02-0.17‰) and in the NHOAc-extractable pool after repeated phytoextraction (ΔCd = 0.07-0.18‰). This indicates different mechanisms of membrane transport (high-affinity transport of Cd) and supplementation of the bioavailable pool in soil (Cd supplied mainly through complexation with root-derived organic ligands) of the two metals. The combination of chemical speciation and stable Zn isotope ratios in the plant and the bioavailable soil pool reveal that the Zn pool related to Fe and Mn oxides became increasingly bioavailable with increasing crop generations. Capsule: Stable isotope analysis indicates that soil Fe- and Mn-oxide bound Zn replenishment boosted Zn uptake by the hyperaccumulator Sedum plumbizincicola during long-term remediation.
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http://dx.doi.org/10.1016/j.scitotenv.2021.151306DOI Listing
February 2022

Virus-templated magnetic composite hydrogels for surface immobilization of mimic-free-lipase.

Nanoscale 2021 Nov 4;13(42):17871-17880. Epub 2021 Nov 4.

Department of Chemical Engineering, Tsinghua University, Beijing 100084, P. R. China.

Surface immobilization of enzymes on magnetic-recoverable carriers is of great interest and importance for the biocatalysis of relatively large molecules. In this work, the nanosized amino-rich filamentous M13 virus, a versatile biological scaffold, was applied as the unique soft backbone for lipase immobilization. Based on the structure and capsid proteins of M13 phages, the magnetic-recoverable mimic-free-lipases (MFLs) composed of the M13 hydrogels and magnetic particles were developed in two designs. In the first design, nanosized wild M13 phages were crosslinked into a phage hydrogel through the N-terminals of pVIII peptides while NH-FeO magnetic nanoparticles (MNPs) were attached to the M13 virus through glutaraldehyde, forming the M13-(NH-FeO) magnetic phage hydrogel. In the second design, special M13 with FeO affinity pIII-peptide (FAP-M13) was biopanned for strongly binding towards bare FeO with the "hook"-like pIII-peptide (N-LPLSTQH-C). TEM observation confirmed the direct grasp of FAP-M13 on bare FeO, forming the magnetic (FAP-M13)-FeO virus hydrogel. Lipases were uniformly anchored on the phage surface of nanoscale by crosslinking with the N-terminals of pVIII peptides, and then [email protected](NH-FeO) and [email protected](FAP-M13)-FeO MFLs were constructed. For both MFLs, high activity recovery yield (>95%) and efficient magnetic separation were characterized. Significantly reduced MNP-usage-amount and enhanced lipase-loading-amount both by about 40 folds were obtained, compared with the conventional NH-FeO carriers. The quantified and / values were almost equal to those of the free lipases, verifying free-enzyme-mimicking features of the MFLs. High pH-tolerance, wide temperature adaptability, enhanced thermal stability and stable magnetic separation capability of both MFLs were also observed. In particular, the (FAP-M13)-FeO magnetic virus hydrogel simply using bare FeO MNPs would be more convenient and economical in the scaled-up biocatalysis.
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http://dx.doi.org/10.1039/d1nr03571aDOI Listing
November 2021

Virus-templated magnetic composite hydrogels for surface immobilization of mimic-free-lipase.

Nanoscale 2021 Nov 4;13(42):17871-17880. Epub 2021 Nov 4.

Department of Chemical Engineering, Tsinghua University, Beijing 100084, P. R. China.

Surface immobilization of enzymes on magnetic-recoverable carriers is of great interest and importance for the biocatalysis of relatively large molecules. In this work, the nanosized amino-rich filamentous M13 virus, a versatile biological scaffold, was applied as the unique soft backbone for lipase immobilization. Based on the structure and capsid proteins of M13 phages, the magnetic-recoverable mimic-free-lipases (MFLs) composed of the M13 hydrogels and magnetic particles were developed in two designs. In the first design, nanosized wild M13 phages were crosslinked into a phage hydrogel through the N-terminals of pVIII peptides while NH-FeO magnetic nanoparticles (MNPs) were attached to the M13 virus through glutaraldehyde, forming the M13-(NH-FeO) magnetic phage hydrogel. In the second design, special M13 with FeO affinity pIII-peptide (FAP-M13) was biopanned for strongly binding towards bare FeO with the "hook"-like pIII-peptide (N-LPLSTQH-C). TEM observation confirmed the direct grasp of FAP-M13 on bare FeO, forming the magnetic (FAP-M13)-FeO virus hydrogel. Lipases were uniformly anchored on the phage surface of nanoscale by crosslinking with the N-terminals of pVIII peptides, and then [email protected](NH-FeO) and [email protected](FAP-M13)-FeO MFLs were constructed. For both MFLs, high activity recovery yield (>95%) and efficient magnetic separation were characterized. Significantly reduced MNP-usage-amount and enhanced lipase-loading-amount both by about 40 folds were obtained, compared with the conventional NH-FeO carriers. The quantified and / values were almost equal to those of the free lipases, verifying free-enzyme-mimicking features of the MFLs. High pH-tolerance, wide temperature adaptability, enhanced thermal stability and stable magnetic separation capability of both MFLs were also observed. In particular, the (FAP-M13)-FeO magnetic virus hydrogel simply using bare FeO MNPs would be more convenient and economical in the scaled-up biocatalysis.
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http://dx.doi.org/10.1039/d1nr03571aDOI Listing
November 2021

A CRISPR/Cas9-based single-stranded DNA recombineering system for genome editing of PD630.

Synth Syst Biotechnol 2021 Sep 10;6(3):200-208. Epub 2021 Aug 10.

Department of Chemical Engineering, Tsinghua University, Beijing, 100084, China.

Genome engineering of PD630, an important microorganism used for the bioconversion of lignin, is currently dependent on inefficient homologous recombination. Although a CRISPR interference procedure for gene repression has previously been developed for PD630, a CRISPR/Cas9 system for gene knockout has yet to be reported for the strain. In this study, we found that the cytotoxicity of Cas9 and the deficiency in pathways for repairing DNA double-strand breaks (DSBs) were the major causes of the failure of conventional CRISPR/Cas9 technologies in , even when augmented with the recombinases Che9c60 and Che9c61. We successfully developed an efficient single-stranded DNA (ssDNA) recombineering system coupled with CRISPR/Cas9 counter-selection, which facilitated rapid and scarless editing of the genome. A two-plasmid system, comprising Cas9 driven by a weak promoter Pniami, designed to prevent cytotoxicity, and a single-guide RNA (sgRNA) under the control of a strong constitutive promoter, was proven to be appropriate with respect to cleavage function. A novel recombinase, RecT derived from a prophage, was identified for the first time, which facilitated recombination of short ssDNA donors (40-80 nt) targeted to the lagging strand and enabled us to obtain a recombination efficiency up to 10-fold higher than that of endogenous pathways. Finally, by incorporating RecT and Cas9 into a single plasmid and then co-transforming cells with sgRNA plasmids and short ssDNA donors, we efficiently achieved gene disruption and base mutation in , with editing efficiencies ranging from 22 % to 100 %. Simultaneous disruption of double genes was also confirmed, although at a lower efficiency. This effective genome editing tool will accelerate the engineering of metabolism.
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http://dx.doi.org/10.1016/j.synbio.2021.08.001DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8365321PMC
September 2021

Chronic Inflammation in Ulcerative Colitis Causes Long-Term Changes in Goblet Cell Function.

Cell Mol Gastroenterol Hepatol 2022 18;13(1):219-232. Epub 2021 Aug 18.

Division of Gastroenterology & Hepatology, Department of Medicine, Baltimore, Maryland.

Background & Aims: One of the features of ulcerative colitis (UC) is a defect in the protective mucus layer. This has been attributed to a reduced number of goblet cells (GCs). However, it is not known whether abnormal GC mucus secretion also contributes to the reduced mucus layer. Our aims were to investigate whether GC secretion was abnormal in UC and exists as a long-term effect of chronic inflammation.

Methods: Colonoids were established from intestinal stem cells of healthy subjects (HS) and patients with UC. Colonoids were maintained as undifferentiated (UD) or induced to differentiate (DF) and studied as three-dimensional or monolayers on Transwell filters. Total RNA was extracted for quantitative real-time polymerase chain reaction analysis. Carbachol and prostaglandin E mediated mucin stimulation was examined by MUC2 IF/confocal microscopy and transmission electron microscopy.

Results: Colonoids from UC patients can be propagated over many passages; however, they exhibit a reduced rate of growth and transepithelial electrical resistance compared with HS. Differentiated UC colonoid monolayers form a thin and non-continuous mucus layer. UC colonoids have increased expression of secretory lineage markers ATOH1 and SPDEF, along with MUC2 positive GCs, but failed to secrete mucin in response to the cholinergic agonist carbachol and prostaglandin E, which caused increased secretion in HS. Exposure to tumor necrosis factor α (5 days) reduced the number of GCs, with a greater percentage decrease in UC colonoids compared with HS.

Conclusions: Chronic inflammation in UC causes long-term changes in GCs, leading to abnormal mucus secretion. This continued defect in GC mucus secretion may contribute to the recurrence in UC.
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http://dx.doi.org/10.1016/j.jcmgh.2021.08.010DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8593611PMC
August 2021

Fingerprinting vanadium in soils based on speciation characteristics and isotope compositions.

Sci Total Environ 2021 Oct 9;791:148240. Epub 2021 Jun 9.

CAS Key Laboratory of Crust-Mantle Materials and Environments, School of Earth and Space Sciences, University of Science and Technology of China, Hefei 230026, Anhui, China; CAS Center for Excellence in Comparative Planetology, China. Electronic address:

Vanadium (V) can have toxic effects on human organs and physiological systems, yet tracing V sources remains challenging. Here, two methods were used for V source tracing in soil based on speciation characteristics and isotope compositions. According to the sequential extraction method of the European Communities Bureau of Reference (BCR), the analysis of speciation distributions offers a possible means of distinguishing V sources. Here, the isotope compositions of polluted soils around a coal-fired power plant and smelter in China were used to identify the sources of V. Significant V isotope variation (δV range = -0.74 ± 0.07; mean ± 2SD = -0.52 ± 0.05‰) was observed in the soil samples, attributed to coal-burning (ΔV = -0.31 ± 0.05‰; mean ± 2SD; n = 1) and smelting processes (ΔV = -0.31 ± 0.07‰; mean ± 2SD; n = 1). All of the soil V isotope ratios plotted within the range of end-member components corresponding to potential V contributors in the environment. Among these, δV ranged from -0.74 ± 0.07 to -0.55 ± 0.02‰ in topsoil, the average δV was -0.52 ± 0.05‰ in the deep soils, and the δV of the end-member components ranged from -0.52 ± 0.05 to -0.94 ± 0.11‰. The primary anthropogenic source of V in the topsoil was fly ash from coal-burning that was consistent with the BCR method results. Furthermore, the downward migration of V was identified in the soil profile adjacent to the smelting plant, and V in the deep soils was dominated by natural sources relative to anthropogenic sources in the surface soils.
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http://dx.doi.org/10.1016/j.scitotenv.2021.148240DOI Listing
October 2021

Analysis of rehabilitation in stroke patients in a High Dependency Unit.

Am J Transl Res 2021 15;13(7):8262-8269. Epub 2021 Jul 15.

Department of Rehabilitation Medicine, Qujing No. 1 Hospital Qujing, Yunnan Province, China.

Objective: To analyze the rehabilitation effict of stroke patients in the High Dependency Unit (HDU).

Methods: A retrospective study was conducted on 96 ischemic stroke patients who met the inclusion criteria and were hospitalized in the Department of Rehabilitation Medicine of Qujing No. 1 Hospital by convenience sampling. All patients were divided into two groups according to the inclusion and exclusion criteria: experimental group (n=49) and control group (n=47). The experimental group was admitted to the HDU and treated with the whole-process of full cycle rehabilitation management and treatment, and the control group was admitted to the general ward and treated with the conventional treatment model of the Department of Rehabilitation Medicine.

Results: The improvement of muscle strength at discharge was compared between the two groups. The results showed that the muscle strength effective rate was 85.7% in the experimental group and 55.3% in the control group, and the difference had statistical significance (P<0.01). The Barthel index score at admission and at discharge were compared between the two groups, the results showed that there was no significant difference in the Barthel index score at admission between the two groups (t=0.668, P>0.05). The Barthel index score at discharge in the experimental group was higher than that in the control group, and the difference had statistical significance (t=7.969, P<0.05). The Montreal Cognitive Assessment (MoCA) score of the experimental group before treatment was (25.67±3.11) points, and the MoCA score after treatment was (29.01±2.21) points; the MoCA score of the control group before treatment was (24.11±4.65) points, and the MoCA score after treatment was (25.35±2.29) points. After implementation of the clinical nursing pathway, the improvement in cognitive function in the experimental group of patients was significantly higher than that in the control group, and the difference had statistical significance (P<0.01). Through the investigation of the satisfaction rate of nursing work in the two groups, it was found that the satisfaction rate in the experimental group was better than that in the control group, and the difference had statistical significance (P<0.05); there was 1 case of pressure sores that occurred in the experimental group, and 3 cases that occurred in the control group, χ=1.133, P=0.287, and the difference had no statistical significance (P>0.05). The length of hospital stay was compared between the two groups, the results showed that the length of hospital stay was (11.76±2.06) days in the experimental group and (14.21±2.40) days in the control group, thus, the average length of hospital stay in the experimental group was less than that in the control group, and the difference had statistical significance (P<0.001).

Conclusion: The whole-process of full cycle rehabilitation management and treatment can improve the activities of daily living, limb muscle strength and cognitive function of patients, as well as shorten the length of hospital stay and improve the satisfaction of patients with treatment. Thus, it is worthy of being widely popularized in clinical application.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8340238PMC
July 2021

Plasma microRNAs can be a potential diagnostic biomarker for endometriosis.

Ginekol Pol 2021 Jun 22. Epub 2021 Jun 22.

HwaMei Hospital, University of Chinese Academy of Sciences, Ningbo, China.

Objectives: Plasma microRNAs are considered potential diagnostic biomarkers for endometriosis. Increasing evidence has shown that a huge number of miRNAs are abnormally expressed in endometriosis plasma and play irreplaceable roles in diagnosis.

Material And Methods: The aim of our study was to identify the differential expression of circular miRNA by reviewing the PubMed, ScienceDirect, and Cochrane databases between normal women and women with endometriosis and analyzing the miRNA data downloaded from the GEO database.

Results: Because of the differential miRNA expression in this review, we evaluated the diagnostic values of the differentially expressed miRNAs, particularly during the menstrual phases. According to the cut-off criteria with |log 2 FC| > 1.0 and P < 0.05, 36 differentially expressed miRNAs were identified, including 13 upregulated miRNAs and 23 downregulated miRNAs. We developed miR-155, miR-574, miR-23a, and miR-520d via a Venn diagram. Functional enrichment analysis considered that the target miRNAs might be involved in various pathways related to endometriosis, including neurotrophin, Hippo, oocyte meiosis, ubiquitin mediated proteolysis, HTLV-Infection, FoxO, and Rap1 signaling pathways. CTNNB1, MYC, and ES R1 of transcription factors were related to the differentially expressed miRNAs.

Conclusions: In summary, our study suggested that a four-miRNA could be included as a prognostic marker in endometriosis.
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http://dx.doi.org/10.5603/GP.a2021.0127DOI Listing
June 2021

Real-Time Tissue Elastography to Evaluate Hepatic Hypoxic-Ischemic Injury Caused by Brain Death.

Ultrasound Q 2021 Jun;37(2):138-143

Department of Ultrasound, Tianjin First Central Hospital, No. 24 Fukang Road, Nankai District, Tianjin, 300192, China.

Abstract: This study aimed to explore the potential of real-time tissue elastography (RTE) in evaluating hepatic hypoxic-ischemic injury caused by brain death. We performed RTE and biopsy for 50 donated liver. Hematoxylin-eosin staining was used to observe hepatocyte acidophilic change. Liver grafts were divided into 2 groups, one nonacidophilic change (n = 7) and the other with acidophilic change (n = 43). Correlation and difference analysis were performed for hematoxylin-eosin staining results and RTE parameters. The result indicated that 4 of the 11 RTE parameters, namely, the area of low strain within the region of interest (%AREA), contrast (CONT), inverse difference moment (IDM), and correlation (CORR) were related to hepatocytes acidophilic change (r = 0.284, P = 0.046; r = 0.349, P = 0.013; r = -0.444, P = 0.001; r = -0.381, P = 0.00). Whereas %AREA and CONT of the nonacidophilic change group were lower than that of the acidophilic change group (P < 0.05), IDM and CORR in nonacidophilic change group were higher than that of the acidophilic change group (P < 0.05); the remaining parameters were not statistically different between 2 groups (P > 0.05). Analysis of receiver operating characteristic curve indicated that the area under the curve of %AREA, CONT, IDM, and CORR were 0.75, 0.79, 0.81, and 0.77, respectively. Based on this, we concluded that the quantitative analysis parameters of RTE could preliminary assess hepatic hypoxic-ischemic injury caused by brain death.
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http://dx.doi.org/10.1097/RUQ.0000000000000497DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8177492PMC
June 2021

Circ_0019435 Exerts Its Functions in the Cellular Process of Cervical Cancer via Epigenetically Silencing DKK1 and PTEN.

Reprod Sci 2021 10 26;28(10):2989-2999. Epub 2021 May 26.

Department of Gynecology, HwaMei Hospital, University of Chinese Academy of Sciences, Ningbo, 315010, Zhejiang, China.

Cervical cancer (CC) is the most serious gynecological malignancy among women worldwide. As a subtype of noncoding RNAs (ncRNAs), circular RNAs (circRNAs) play important roles in the regulation of gene expression and cancer progression. It was discovered from the cancer-specific circRNA database (CSCD) that circ_0019435 was mainly distributed in the nucleus of HeLa-S3 cells. However, few researches have mentioned circ_0019435 with its function in cancers. The present study uncovered that circ_0019435 was upregulated in CC cells by qRT-PCR. Moreover, circ_0019435 was more stable than its linear isoform-ABCC2. Besides, no regulation of circ_0019435 on ABCC2 and the chemoresistance of CC cells were found. Then, it was unveiled by a series of functional assays including colony formation, trypan blue staining, and transwell invasion assays in that circ_0019435 ablation induced the suppression of proliferation, invasion, and EMT of HeLa and SiHa cells. The subcellular distribution of circ_0019435 was assessed by subcellular fractionation and FISH assay. Furthermore, it was disclosed that circ_0019435 binds to EZH2 to silence DKK1 and PTEN. Finally, rescue assays corroborated that DKK1 and PTEN were involved in circ_0019435-mediated CC cell progression. In conclusion, circ_0019435 regulates DKK1 and PTEN expression at the epigenetic level, thereby influencing the progression of CC cells.
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http://dx.doi.org/10.1007/s43032-021-00625-zDOI Listing
October 2021

Clinical Strains of With Strong Cell Invasiveness and the Protective Effect of Patchouli Alcohol by Improving miR-30b/C Mediated Xenophagy.

Front Pharmacol 2021 30;12:666903. Epub 2021 Apr 30.

Shenzhen Traditional Chinese Medicine Hospital, the Fourth Clinical Medical College of Guangzhou University of Chinese Medicine, Shenzhen, China.

was classified by the World Health Organization as a class 1 carcinogen. The development of drug-resistant strains of this pathogen poses a serious threat to human health worldwide. The cell invasion of activates xenophagy in gastric epithelial cells by mediating miR-30b/c, and the emergence of autophagosomes provides a niche that enables the survival of intracellular and promotes its drug resistance. This study revealed that some clinical drug-resistant strains present much stronger invasive ability than standard strains. Patchouli alcohol (PA), a tricyclic sesquiterpene from (Blanco) Benth (Labiatae), showed reliable activity against intracellular . The mechanisms appeared to involve the downregulation of miR-30c-3p/5p and miR-30b-5p, thereby upregulating xenophagy-related gene expression (ULK1, ATG5, ATG12, and ATG14) and enhancing xenophagy. PA also inhibited the nuclear transfection of miR-30b-5p induced by , thereby enhancing transcription factor EB function and increasing lysosome activity. The finding of strongly invasive intracellular has great implications for clinical treatment, and PA can act against invasive based on the improvement of miR-30b/c mediated xenophagy. Taken together, the results demonstrate that PA have potential use as a candidate medication for intracellular drug-resistant .
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http://dx.doi.org/10.3389/fphar.2021.666903DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8120110PMC
April 2021

Equity analysis of Chinese physician allocation based on Gini coefficient and Theil index.

BMC Health Serv Res 2021 May 12;21(1):455. Epub 2021 May 12.

School of Economics and Management, Jiangxi University of Traditional Chinese Medicine, Nanchang, 330000, Jiangxi, China.

Background: Unequal allocation of medical physician resource represents one of major problems in the current medical service management in China and many other countries. This study is designed to analyze the current distribution of physicians in 31 provincial administrative regions in China, to estimate the fairness of the distribution of physicians and provide a theoretical basis for the improvement of the allocation of physicians.

Methods: This study took physicians from 31 provincial administrative regions in China as the study objects, and the data were obtained from the China Health Statistics Yearbook 2019 and the official website of the National Bureau of Statistics of China. Calculation of the Gini coefficient (G) and the Theil index (T) were carried out by drawing the Lorenz curve. The fairness of present physician location in 31 provincial administrative regions in China was analyzed from the perspective of distribution by both population and service area.

Results: The Gini coefficients of medical physicians in China are 0.003 and 0.88 by population and by service area, respectively. This shows that the distribution of medical physicians is fair basing on population, and there is little difference in the number of physicians per 1000 population in different regions. However, the physician distribution basing on service area is highly unfair and shows a large gap in the number of physicians per square kilometer between different regions. In general, Beijing, Zhejiang, Shanghai, Jiangsu, Shandong, and Tianjin are higher than the overall level of 31 provincial administrative regions. In addition, the number of medical physicians in Zhejiang, Shandong, Beijing and Jiangsu is over-provisioned.

Conclusion: Bridging the number of medical physicians in different regions is a key step to improve the equity of physicians' resource allocation. Thus, findings from this study emphasize the need to take more measures to reduce physician quality differences between regions, balance and coordinate medical resources. This will increase the access of all citizens to quality medical services.
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http://dx.doi.org/10.1186/s12913-021-06348-wDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8115393PMC
May 2021

Genetic toolkits for engineering Rhodococcus species with versatile applications.

Biotechnol Adv 2021 Jul-Aug;49:107748. Epub 2021 Apr 3.

Department of Chemical Engineering, Tsinghua University, Beijing 100084, China; Key Laboratory of Industrial Biocatalysis (Tsinghua University), the Ministry of Education, Beijing 100084, China; Center for Synthetic and Systems Biology, Tsinghua University, Beijing 100084, China. Electronic address:

Rhodococcus spp. are a group of non-model gram-positive bacteria with diverse catabolic activities and strong adaptive capabilities, which enable their wide application in whole-cell biocatalysis, environmental bioremediation, and lignocellulosic biomass conversion. Compared with model microorganisms, the engineering of Rhodococcus is challenging because of the lack of universal molecular tools, high genome GC content (61% ~ 71%), and low transformation and recombination efficiencies. Nevertheless, because of the high interest in Rhodococcus species for bioproduction, various genetic elements and engineering tools have been recently developed for Rhodococcus spp., including R. opacus, R. jostii, R. ruber, and R. erythropolis, leading to the expansion of the genetic toolkits for Rhodococcus engineering. In this article, we provide a comprehensive review of the important developed genetic elements for Rhodococcus, including shuttle vectors, promoters, antibiotic markers, ribosome binding sites, and reporter genes. In addition, we also summarize gene transfer techniques and strategies to improve transformation efficiency, as well as random and precise genome editing tools available for Rhodococcus, including transposition, homologous recombination, recombineering, and CRISPR/Cas9. We conclude by discussing future trends in Rhodococcus engineering. We expect that more synthetic and systems biology tools (such as multiplex genome editing, dynamic regulation, and genome-scale metabolic models) will be adapted and optimized for Rhodococcus.
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http://dx.doi.org/10.1016/j.biotechadv.2021.107748DOI Listing
June 2021

Serum-Derived Exosomes from Patients with Coronary Artery Disease Induce Endothelial Injury and Inflammation in Human Umbilical Vein Endothelial Cells.

Int Heart J 2021 Mar 17;62(2):396-406. Epub 2021 Mar 17.

Second School of Clinical Medicine, Southern Medical University.

Endothelial injury and inflammation have been found to be essential in the pathogenesis of coronary artery disease (CAD). Circulating exosomes are of great value as novel biomarkers for CAD. However, the role of circulating exosomes in the pathogenesis of CAD remains unclear. Thus, in this study, we aimed to examine whether circulating exosomes from CAD are involved in the endothelial injury and inflammation. The serum-derived exosomes were isolated from CAD and controls using an ExoQuick reagent, and these were then quantified by measuring the protein levels using BCA methods. The uptake of exosomes by human umbilical vein endothelial cells (HUVECs) was observed by laser scanning microscope and analyzed via flow cytometry. Then, HUVECs were treated with vehicle, exosomes from CAD (CAD-exo), and controls (ctrl-exo) in the absence and presence of vascular endothelial growth factor (VEGF). Cell viability, migration, and angiogenesis were evaluated using CCK-8 assay, scratch assay, and tube formation assay. Inflammatory factors including IL-1β, IL-6, TNF-α, ICAM-1, and VCAM-1 levels were detected via qPCR. As per our findings, no significant differences were noted in uptake of ctrl-exo and CAD-exo by HUVECs. CAD-exo suppressed cell viability in a dose-dependent manner. Compared with ctrl-exo, CAD-exo-treated HUVECs significantly suppressed migration and angiogenesis. However, CAD-exo had a stronger inhibitory effect on VEGF-induced migration and angiogenesis compared with ctrl-exo. Moreover, IL-1β, TNF-α, and ICAM-1 were determined to be significantly upregulated in HUVECs treated with CAD-exo, but IL-6 and VCAM-1 expressions were not affected. Overall, our results suggest that CAD-exo are involved in endothelial injury and inflammation, which may, in turn, cause endothelial dysfunction and potentially promote the development of CAD.
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http://dx.doi.org/10.1536/ihj.20-641DOI Listing
March 2021

Intravenous immunoglobulin G in the treatment of ABO hemolytic disease of the newborn during the early neonatal period at a tertiary academic hospital: a retrospective study.

J Perinatol 2021 06 15;41(6):1397-1402. Epub 2021 Feb 15.

Department of Neonatology, the Children's Hospital, Zhejiang University School of Medicine, National Clinical Research Center for Child Health, Hangzhou, Zhejiang, People's Republic of China.

Objective: To evaluate the efficacy and safety of intravenous immunoglobulin G (IVIG) in infants with ABO hemolytic disease of the newborn (HDN).

Methods: Infants with moderate-to-severe ABO HDN during early neonatal period (<7 days) at our hospital in 2017 were included in this retrospective study. Patients treated with IVIG and phototherapy were classified as the IVIG group, and those who only received phototherapy were classified as the phototherapy only group.

Results: Forty-six patients were classified into the IVIG group and 68 other patients were classified into the phototherapy only group. There was no significant difference in duration of phototherapy, hospitalization periods, needs for exchange transfusion, transfusions, and incidence of bilirubin-induced neurological sequelae between these two groups (P = 0.20, 0.27, 0.65, 0.47, 0.78, respectively).

Conclusion: It seems unnecessary to expose neonates to IVIG in moderate-to-severe ABO HDN when the available data show no appreciable benefits.
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http://dx.doi.org/10.1038/s41372-021-00963-5DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8225508PMC
June 2021

Optimization of Cephalosporin C Acylase Expression in Escherichia coli by High-Throughput Screening a Constitutive Promoter Mutant library.

Appl Biochem Biotechnol 2021 Apr 6;193(4):1056-1071. Epub 2021 Jan 6.

Department of Chemical Engineering, Tsinghua University, Beijing, 100084, China.

Cephalosporin C acylase (CCA) is capable of catalyzing cephalosporin C (CPC) to produce 7-aminocephalosporanic acid (7-ACA), an intermediate of semi-synthetic cephalosporins. Inducible expression is usually used for CCA. To improve the efficiency of CCA expression without gene induction, three recombinant strains regulated by constitutive promoters BBa_J23105, PLtetO1, and tac were constructed, respectively. Among them, BBa_J23105 was the best promoter and its mutant libraries were established using saturation mutagenesis. In order to obtain the mutants with enhanced activity, a high-throughput screening method based on flow cytometric sorting techniques was developed by using green fluorescent protein (GFP) as the reporter gene. A series of mutants were screened at 28 °C, 200 rpm, and 24-h culture condition. The study of mutants showed that the enzyme activity, fluorescence intensity, and promoter transcriptional strength were positively correlated. The enzyme activity of the optimal mutant obtained by screening reached 12772 U/L, 3.47 times that of the original strain.
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http://dx.doi.org/10.1007/s12010-020-03482-9DOI Listing
April 2021

Evaluation of factors affecting alveolar ridge height and facial bone thickness in Chinese maxillary central incisors by cone beam CT.

J Dent Sci 2021 Jan 8;16(1):229-235. Epub 2020 Jun 8.

Department of Implantology, Stomatological Hospital, Southern Medical University, Guangzhou, 510280, China.

Background/purpose: In the immediate implantation of maxillary central incisors, the height of the alveolar bone is lost, and there is often a risk of bone fracture due to the thin buccal bone wall (BBW). The purpose of this study was to assess the effects of smoking, age, and root position in the alveolar bone on the BBW and the distance between the cemento-enamel junction (CEJ) and the facial bone crest (FBC) of Chinese maxillary central incisors.

Materials And Methods: The patients were divided by smoking, gender, age, and root sagittal position in the alveolar bone. BBW thickness was measured at the following sites: the 4 mm apical to the CEJ, the middle of the root, and the apex. The distance from the CEJ to the FBC was also evaluated.

Results: Cone beam CT (CBCT) data for the maxillary central incisors of 645 patients (323 males and 322 females) were selected and analyzed. The CEJ-FBC distance in patients who smoked (2.79 ± 0.78 mm) was significantly greater than that of non-smokers (2.54 ± 0.69 mm). The BBW in subtype III (0.74 ± 0.43 mm, 0.81 ± 0.36 mm) was thinner than that in subtypes I and II at 4 mm apical to the CEJ and in the middle of the root, with a statistically significant difference ( < 0.05).

Conclusion: In most Chinese people, smoking, gender, age, and the position of the root in alveolar bone are all important factors that must be considered before immediate implantation is undertaken.
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http://dx.doi.org/10.1016/j.jds.2020.05.021DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7770285PMC
January 2021

Circulating Exosomal miRNAs as Novel Biomarkers for Stable Coronary Artery Disease.

Biomed Res Int 2020 11;2020:3593962. Epub 2020 Dec 11.

The Second School of Clinical Medicine, Southern Medical University, Guangzhou, Guangdong 510515, China.

Exosomal miRNAs are currently being explored as a novel class of biomarkers in cardiovascular diseases. However, few reports have focused on the value of circulating exosomal miRNAs as biomarkers for stable coronary artery disease (SCAD). Here, we aimed to investigate whether miRNAs involved in cardiovascular diseases in circulating exosomes could serve as novel diagnostic biomarkers for SCAD. Firstly, the serum exosomes were isolated and purified by the ExoQuick reagent and identified by transmission electron microscopy, western blot, and nanoparticle tracking analysis. Then, the purified exosomes were quantified by measuring the exosome protein concentration and calculating the total protein amount. Next, eight miRNAs involved in cardiovascular diseases, miR-192-5p, miR-148b-3p, miR-125a-3p, miR-942-5p, miR-149-5p, miR-32-5p, miR-144-3p, and miR-142-5p, were quantified in circulating exosomes from the control group ( = 20) and the SCAD group ( = 20) by quantitative real-time polymerase chain reaction (qPCR). Finally, the gene targets of the differentially expressed miRNAs were predicted, and the functions and signaling pathways of these targets were analyzed using an online database. The isolated exosomes had a bilayer membrane with a diameter of about 100 nm and expressed exosomal markers including CD63, Tsg101, and Flotillin but negatively expressed Calnexin. Both the exosome protein concentration and total protein amount exhibited no significant differences between the two groups. The qPCR assay demonstrated that among the eight miRNAs, the expression levels of miR-942-5p, miR-149-5p, and miR-32-5p in the serum exosomes from the SCAD group were significantly higher than that from the control group. And the three miRNAs for SCAD diagnosis exhibited AUC values of 0.693, 0.702, and 0.691, respectively. GO categories and signaling pathways analysis showed that some of the predictive targets of these miRNAs were involved in the pathophysiology processes of SCAD. In conclusion, our findings suggest that serum exosomal miR-942-5p, miR-149-5p, and miR-32-5p may serve as potential diagnostic biomarkers for SCAD.
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http://dx.doi.org/10.1155/2020/3593962DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7748912PMC
June 2021

[Construction and optimization of engineered Bacillus subtilis for surfactin production].

Sheng Wu Gong Cheng Xue Bao 2020 Nov;36(11):2377-2386

Daqing Refining & Petrochemical Company, Daqing 163000, Heilongjiang, China.

Surfactin has great potential applications in enhancing oil recovery, agriculture, pharmaceuticals, foods and beverages, and cosmetics due to its extraordinary surface activity, biodegradability, anti-bacterial activity and biocompatibility. Enhancing surfactin production by engineering surfactin-producer and optimizing culture conditions is the key of its industrial production and subsequent applications. In this study, the effect of fatty acid synthesis pathway on surfactin synthesis was investigated, and Bacillus subtilis THBS-2 and THBS-8 with high surfactin titer were constructed by overexpressing key genes involved in the fatty acid synthesis pathway. To optimize culture condition, the amount and adding time of isopropyl-beta-D-thiogalactopyranoside (IPTG) and amino acids were studied, and a two-stage culture method was obtained: IPTG (final concentration: 1.25 mmol/L) and leucine (final concentration: 5 g/L) were added at 3 h, leucine (final concentration 5 g/L) and condensed culture medium (5 mL) were added at 24 h. Applying this strategy, the surfactin titer of B. subtilis THBS-2 reached to 24 g/L in shake flask at 48 h and up to 34 g/L after 68 h fermentation in a 30-L fermentor. The results provide basis for large-scale production and broad application of surfactin.
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http://dx.doi.org/10.13345/j.cjb.200156DOI Listing
November 2020

Jian-Pi-Yi-Shen Regulates EPO and Iron Recycling Protein Expressions in Anemic Rats with Chronic Kidney Disease: Accumulation of Hypoxia Inducible Factor-2 via ERK Signaling.

Evid Based Complement Alternat Med 2020 30;2020:8894257. Epub 2020 Oct 30.

The Fourth Clinical Medical College of Guangzhou University of Chinese Medicine, Shenzhen 518033, China.

Jian-Pi-Yi-Shen (JPYS), the traditional Chinese medicine (TCM) decoction, has been commonly used to treat chronic kidney disease (CKD) and its complications such as anemia. JPYS has been previously found to induce erythropoietin (EPO) production in HEK293T cells and CKD rats. However, the mechanism of JPYS in treating anemia of CKD rats has remained largely unknown. Here, we further extend our effort to investigate the translational control of hypoxia inducible factor- (HIF-) protein via ERK signaling and the effect on iron recycling-related protein expression by JPYS, thus revealing the mechanism of JPYS in correcting anemia in CKD. Experimental CKD rats with anemia were induced by 5/6 nephrectomy. Rats were administrated orally with high dose (6.0 g/kg/d) and low dose (1.5 g/kg/d) of JPYS for 90 days. Serum hepcidin level was determined to evaluate iron homeostasis. The protein expressions of HIF-2, erythropoietin (EPO), ferritin, and ferroportin (FPN) and the phosphorylation level of extracellular signal-regulated kinase 1/2 (ERK1/2) were detected by Western blot. The results showed that JPYS treatment significantly ameliorated kidney function by reducing increased levels of blood urea nitrogen (BUN), serum creatinine (Scr), and urine protein (UPRO). Periodic acid-Schiff (PAS) and Masson staining observation showed that the renal pathological damage was restored in JPYS-treated CKD rats. In parallel, JPYS markedly improved CKD anemia through upregulation of red blood cell (RBC), hemoglobin (HGB), and hematocrit (HCT). JPYS stimulated EPO and HIF-2 protein expressions in both the kidney and liver of CKD rats. Furthermore, JPYS induced the phosphorylation of ERK1/2 protein. In addition, JPYS regulated protein expression of ferritin and FPN in both the liver and spleen of CKD rats and the serum level of hepcidin. In conclusion, JPYS induces the expression of EPO through ERK-mediated HIF-2 protein accumulation and regulates systemic iron recycling, supporting its role in promoting erythropoiesis and improvement of anemia in CKD.
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http://dx.doi.org/10.1155/2020/8894257DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7647762PMC
October 2020

Pediatric living donor left lateral segment liver transplantation for biliary atresia: Doppler ultrasound findings in early postoperative period.

Jpn J Radiol 2021 Apr 8;39(4):367-375. Epub 2020 Nov 8.

Department of Ultrasound, Tianjin First Central Hospital, No. 24 Fukang Road, Nankai District, Tianjin, 300192, China.

Purpose: To analyze hepatic hemodynamic parameters detected by Doppler ultrasound (DU) of uncomplicated children with biliary atresia who underwent left lateral segment living donor liver transplantation (LLS-LDLT), explore its normal change trend over time and determine the normal reference interval.

Methods: We retrospectively involved the data from 227 biliary atresia patients (100 Males,127 Females). Hemodynamic parameters include peak systolic velocity (PSV), end-diastolic velocity (EDV), resistivity index (RI), and pulsation index (PI) of the hepatic artery (HA), portal vein velocity (PVV), portal vein flow (PVF) and hepatic vein velocity (HVV) during intra-operative and on the 1st, 3rd, 5th and 7th day after operation were collected. Repeated measures analysis of the variance and Friedman test were used to analyze the changing trend of hemodynamic parameters over time in the first week after the operation.

Results: PSV and EDV showed a similar changing tendency at one week after surgery, with an overall decrease-rise trend; RI and PI also changed similarly with an overall rise-decrease trend. The HVV and PVV at surgery were lower than at all time points after surgery. As for PVF, the value of POD5 was the highest and then decreased. Additionally, this study provided the normal reference interval of hemodynamic parameters for LLS-LDLT patients, which were PSV: 18.4-98.3 cm/s, EDV: 0-43.3 cm/s, RI: 0.41-1.0, PI: 0.51-2.0, PVV: 19.0-83.7 cm/s, HVV: 19.4-68.0 cm/s, and PVF:99.5-500.0 ml/min/100 g at intraoperation. Within the first postoperative week: PSV: 21.0-97.7 cm/s, EDV: 0-32.7 cm/s, RI: 0.47-1.0, PI: 0.62-2.0, PVV: 23.0-92.0 cm/s, HVV: 19.7-86.0 cm/s, and PVF: 100.0-513.0 ml/min/100 g.

Conclusion: The hepatic hemodynamic of post-transplanted children detected by DU had specific changing trends and normal ranges, which provides valuable reference values for ultrasonologists and pediatric transplant clinicians.
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http://dx.doi.org/10.1007/s11604-020-01067-4DOI Listing
April 2021

Distribution and gas-particle partitioning of polycyclic aromatic hydrocarbons over the East China Sea and Yellow Sea in spring: Role of atmospheric transport transition.

Sci Total Environ 2021 Mar 17;762:143071. Epub 2020 Oct 17.

Shanghai Key Laboratory of Atmospheric Particle Pollution and Prevention, Institute of Atmospheric Sciences, Department of Environmental Science and Engineering, Fudan University, Shanghai 200433, China; Institute of Eco-Chongming (IEC), Shanghai 200062, China. Electronic address:

To elucidate the variations in the East Asian monsoon system during seasonal changes and their impacts on continental outflow of polycyclic aromatic hydrocarbons (PAHs), sixteen integrated air samples were collected during a research cruise covering the Yellow Sea (YS) and East China Sea (ECS) in mid-spring of 2017. The concentrations of total suspended particle (TSP), aerosol-phase PAH fractions, ratios of organic to elemental carbon (OC/EC) and gas-particle partitioning of atmospheric PAHs exhibited clear regional differences associated with variations in the monsoon regime. The total concentrations of 16 USEPA priority PAHs (ΣPAHs) varied from 3.11 to 13.4 ng/m throughout the cruise, with medium-to-high molecular weight (MW) PAHs more enriched over the YS and north ECS than the south ECS. Together with the relatively low gaseous PAH fraction over the YS and north ECS (78 ± 4%) relative to the south ECS (95 ± 13%), this result indicates the pattern of regional atmospheric transport. The ratio of organic to elemental carbon varied significantly between the south ECS (lower than 4) and the YS and north ECS (greater than 4), indicating contributions from vehicle emissions and coal combustion or biomass burning, respectively, following different atmospheric input pathways of carbonaceous aerosols, as supported by backward trajectory analysis. Considering the gas-particle partitioning of PAHs, soot adsorption was the main partitioning mechanism in the study region; while high-MW PAHs in the YS and north ECS were influenced by both absorption and adsorption. The K absorption model provided better predictions for high-MW PAHs when continental air masses prevailed, despite underestimating the partition coefficients (k) of low-MW PAHs. Meanwhile, predicted k for medium MW PAHs was better estimated over the YS and ECS when K was included.
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http://dx.doi.org/10.1016/j.scitotenv.2020.143071DOI Listing
March 2021

Fluid shear stress enhances differentiation of jejunal human enteroids in Intestine-Chip.

Am J Physiol Gastrointest Liver Physiol 2021 03 14;320(3):G258-G271. Epub 2020 Oct 14.

Department of Medicine, Division of Gastroenterology and Hepatology, Johns Hopkins University School of Medicine, Baltimore, Maryland.

There is increasing evidence that the study of normal human enteroids duplicates many known aspects of human intestinal physiology. However, this epithelial cell-only model lacks the many nonepithelial intestinal cells present in the gastrointestinal tract and exposure to the mechanical forces to which the intestine is exposed. We tested the hypothesis that physical shear forces produced by luminal and blood flow would provide an intestinal model more closely resembling normal human jejunum. Jejunal enteroid monolayers were studied in the Emulate, Inc. Intestine-Chip under conditions of constant luminal and basolateral flow that was designed to mimic normal intestinal fluid flow, with human umbilical vein endothelial cells (HUVECs) on the basolateral surface and with Wnt3A, R-spondin, and Noggin only on the luminal surface. The jejunal enteroids formed monolayers that remained confluent for 6-8 days, began differentiating at least as early as post plating, and demonstrated continuing differentiation over the entire time of the study, as shown by quantitative real-time polymerase chain reaction and Western blot analysis. Differentiation impacted villus genes and proteins differently with early expression of regenerating family member 1α (REG1A), early reduction to a low but constant level of expression of Na-K-2Cl cotransporter 1 (NKCC1), and increasing expression of sucrase-isomaltase (SI) and downregulated in adenoma (DRA). These results were consistent with continual differentiation, as was shown to occur in mouse villus enterocytes. Compared with differentiated enteroid monolayers grown on Transwell inserts, enteroids exposed to flow were more differentiated but exhibited increased apoptosis and reduced carbohydrate metabolism, as shown by proteomic analysis. This study of human jejunal enteroids-on-chip suggests that luminal and basolateral flow produce a model of continual differentiation over time and NaCl absorption that mimics normal intestine and should provide new insights in intestinal physiology. This study showed that polarized enteroid models in which there is no basolateral Wnt3a, are differentiated, regardless of the Wnt3a status of the apical media. The study supports the concept that in the human intestine villus differentiation is not an all or none phenomenon, demonstrating that at different days after lack of basolateral Wnt exposure, clusters of genes and proteins exist geographically along the villus with different domains having different functions.
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http://dx.doi.org/10.1152/ajpgi.00282.2020DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8202237PMC
March 2021

Enhancing single-cell hyaluronic acid biosynthesis by microbial morphology engineering.

Synth Syst Biotechnol 2020 Dec 30;5(4):316-323. Epub 2020 Sep 30.

Key Laboratory of Industrial Biocatalysis, Ministry of Education, Beijing, 100084, PR China.

Microbial morphology engineering is a novel approach for cell factory to improve the titer of target product in bio-manufacture. Hyaluronic acid (HA), a valuable glycosaminoglycan polymerized by HA synthase (HAS), a membrane protein, is particularly selected as the model product to improve its single-cell HA-producing capacity via morphology engineering. DivIVA and FtsZ, the cell-elongation and cell division related protein, respectively, were both down/up dual regulated in via weak promoter substitution or plasmid overexpression. Different from the natural short-rod shape, varied morphologies of engineered cells, i.e. small-ellipsoid-like (DivIVA-reduced), bulb-like (DivIVA-enhanced), long-rod (FtsZ-reduced) and dumbbell-like (FtsZ-enhanced), were observed. Applying these morphology-changed cells as hosts for HA production, the reduced expression of both DivIVA and FtsZ seriously inhibited normal cell growth; meanwhile, overexpression of DivIVA didn't show morphology changes, but overexpression of FtsZ surprisingly change the cell-shape into long and thick rod with remarkably enlarged single-cell surface area (more than 5.2-fold-increase). And finally, the single-cell HA-producing capacity of the FtsZ-overexpressed was immensely improved by 13.5-folds. Flow cytometry analyses verified that the single-cell HAS amount on membrane was enhanced by 2.1 folds. This work is pretty valuable for high titer synthesis of diverse metabolic products with microbial cell factory.
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http://dx.doi.org/10.1016/j.synbio.2020.09.002DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7530263PMC
December 2020

Single-gene regulated non-spore-forming Bacillus subtilis: Construction, transcriptome responses, and applications for producing enzymes and surfactin.

Metab Eng 2020 11 21;62:235-248. Epub 2020 Aug 21.

Key Laboratory of Industrial Biocatalysis, Ministry of Education, China.

Bacillus subtilis, a spore-forming industrial bacterium, is widely used for production of enzymes and valuable chemicals. The spore-formation, however, always results in remarkably reduced cell-density, thereby reducing product yield. Here, we constructed different non-spore-forming B. subtilis mutants via single-gene regulation. During the three spore-forming stages: signal sensing, transduction, and sporulation, we found that deleting only a single gene of sporulation, i.e. spo0A, spoIIIE, and spoIVB, can completely block the spore generation. Interestingly, the engineered non-sporulating mutants exhibited physiological heterogeneity and distinct synthetic capabilities. The spo0A-null spore-free mutant displayed remarkably high enzyme production capacity, such as 194% enhance amylase production. However, the spoIVB-null non-spore-forming mutant was especially efficient in producing secondary metabolites, such as surfactin; its flask titer increased significantly to 16.7 g/L, with the overexpression and Leu addition strategy. Our results offer a new strategy for re-modeling B. subtilis to further improve its fermentation efficiency and application.
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http://dx.doi.org/10.1016/j.ymben.2020.08.008DOI Listing
November 2020
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