Publications by authors named "Shuangshuang Wang"

98 Publications

Clinical Application of Vacuum Sealing Drainage for the Treatment of Deep Burn Wounds.

Am Surg 2021 Nov 19:31348211054527. Epub 2021 Nov 19.

Department of Burns and Plastic Surgery, The First People's Hospital of Wenling, Wenling, China.

Background: This study aimed to determine the effect of the clinical application of vacuum sealing drainage (VSD) on the treatment of deep burn wounds.

Methods: This single-blind, randomized, controlled study included patients who were admitted to our hospital with deep burns from January 2018 to December 2020; the patients were randomly divided into the VSD and control (CON) groups. The number of days from treatment to skin grafting; survival rate of the first skin graft; rate of positive bacterial culture; visual analog scale (VAS) pain score; and durations of wound healing, antibiotic drug use, and hospitalization were analyzed and compared between the groups.

Results: The application of VSD significantly shortened the number of days from treatment to skin grafting ( < .05); improved the survival rate of the first skin graft in patients with severe burns ( < .05); reduced the rate of positive bacterial culture ( < .05); reduced the VAS pain score ( < .05); and shortened the durations of wound healing ( < .05), antibiotic drug use ( < .05), and hospitalization ( < .05).

Conclusion: Vacuum sealing drainage had a good clinical effect on the recovery of deep burn wounds.
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http://dx.doi.org/10.1177/00031348211054527DOI Listing
November 2021

Digital Numbers Constructed by Fine Patterned Polydopamine on DNA Templates.

Macromol Rapid Commun 2021 Nov 2;42(21):e2100441. Epub 2021 Sep 2.

Hubei Key Laboratory of Bioinorganic Chemistry and Materia Medica, Hubei Engineering Research Center for Biomaterials and Medical Protective Materials, School of Chemistry and Chemical Engineering, Huazhong University of Science and Technology, Luoyu Road 1037, Hongshan, Wuhan, 430074, P. R. China.

Using DNA nanostructures as templates to synthesize shape-controlled polydopamine (PDA) is a promising strategy to realize the fabrication of exquisite PDA nanomaterials. However, previous studies using small DNA tiles as templates could only afford very simple structures such as lines and crosses due to the limited space on the template and the relatively low resolution of the PDA nanopatterns. Therefore, the best resolution of the PDA nanostructures that can be achieved by this technique is carefully investigated. And by connecting several DNA tiles together, larger DNA templates are built up and achieve the synthesis of complicated digital nanopatterned PDA structures.
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http://dx.doi.org/10.1002/marc.202100441DOI Listing
November 2021

Inhibition of Calcineurin/NFAT Signaling Blocks Oncogenic H-Ras Induced Autophagy in Primary Human Keratinocytes.

Front Cell Dev Biol 2021 19;9:720111. Epub 2021 Jul 19.

Department of Tissue Engineering and Regeneration, School and Hospital of Stomatology, Cheeloo College of Medicine, Shandong University & Shandong Key Laboratory of Oral Tissue Regeneration and Shandong Engineering Laboratory for Dental Materials and Oral Tissue Regeneration, Jinan, China.

Mutations of H-Ras, a member of the RAS family, are preferentially found in cutaneous squamous cell carcinomas (SCCs). H-Ras has been reported to induce autophagy, which plays an essential role in tissue homeostasis in multiple types of cancer cells and in fibroblasts, however, the potential role of H-Ras in regulating autophagy in human keratinocytes has not been reported. In this study, we found that the stable expression of the G12V mutant of H-RAS (H-Ras ) induced autophagy in human keratinocytes, and interestingly, the induction of autophagy was strongly blocked by inhibiting the calcineurin/nuclear factor of activated T cells (NFAT) pathway with either a calcineurin inhibitor (Cyclosporin A) or a NFAT inhibitor (VIVIT), or by the small interfering RNA (siRNA) mediated knockdown of calcineurin B1 or NFATc1 , as well as To characterize the role of the calcineurin/NFAT pathway in H-Ras induced autophagy, we found that H-Ras promoted the nuclear translocation of NFATc1, an indication of the activation of the calcineurin/NFAT pathway, in human keratinocytes. However, activation of NFATc1 either by the forced expression of NFATc1 or by treatment with phenformin, an AMPK activator, did not increase the formation of autophagy in human keratinocytes. Further study revealed that inhibiting the calcineurin/NFAT pathway actually suppressed H-Ras expression in H-Ras overexpressing cells. Finally, chromatin immunoprecipitation (ChIP) assays showed that NFATc1 potentially binds the promoter region of H-Ras and the binding efficiency was significantly enhanced by the overexpression of H-Ras , which was abolished by treatment with the calcineurin/NFAT pathway inhibitors cyclosporine A (CsA) or VIVIT. Taking these data together, the present study demonstrates that the calcineurin/NFAT signaling pathway controls H-Ras expression and interacts with the H-Ras pathway, involving the regulation of H-Ras induced autophagy in human keratinocytes.
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http://dx.doi.org/10.3389/fcell.2021.720111DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8328491PMC
July 2021

Developmental validation of the Microreader™ RM-Y ID System: a new rapidly mutating Y-STR 17-plex system for forensic application.

Int J Legal Med 2021 Jul 24. Epub 2021 Jul 24.

Department of Forensic Genetics, West China School of Basic Medical Sciences & Forensic Medicine, Sichuan University, 3-16 Renmin South Road, Chengdu, 610041, China.

Y-chromosomal short tandem repeats (Y-STRs) are widely applied to evolutionary, genealogical, and kinship analyses of male linages in forensic studies, but these low to midrange mutated Y-STRs typically fail to separate related males from the same paternal lineage. Recently, rapidly mutating Y-STRs (RM Y-STRs) have been demonstrated to improve the differentiation of male relatives and individuals. The Microreader™ RM-Y ID System is a new RM Y-STR kit that is capable of simultaneously amplifying 17 RM Y-STRs. Herein, to verify the efficiency and accuracy of the Microreader™ RM-Y ID System, developmental validation was conducted, including PCR-based studies, sensitivity, stability, species specificity, mixture, stutter percentage, and precision studies. Full profiles could be obtained when the hematin concentration was 250 μM, humic acid concentration was 1500 ng/μl, and tannic acid concentration was 200 ng/μl. Full profiles of the mixture of males/males could be detected up to a ratio of 19:1, and full profiles of females/males could always be detected even at ratios up to 24,000:1. Moreover, the forensic characteristics of 250 DNA-confirmed father-son pairs were analysed. The results showed that these 17 RM Y-STRs had high power for forensic discrimination (HD = 1) in the Chinese Han population, and the mutation rates were in the range of 4 × 10 (95% CI 1.00 × 10 to 2.21 × 10, DYS464) to 8.8 × 10 (95% CI 5.60 × 10 to 1.30 × 10, DYF399S1), indicating that the kit was effective for RM Y-STR studies and absolute individualisation of interrelated male individuals.
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http://dx.doi.org/10.1007/s00414-021-02632-wDOI Listing
July 2021

The Changes of T-Wave Amplitude and QT Interval Between the Supine and Orthostatic Electrocardiogram in Children With Dilated Cardiomyopathy.

Front Pediatr 2021 6;9:680923. Epub 2021 Jul 6.

Department of Pediatric Cardiovasology, Children's Medical Center, The Second Xiangya Hospital, Central South University, Changsha, China.

Electrocardiogram (ECG) can be affected by autonomic nerves with body position changes. The study aims to explore the ECG changes of children with dilated cardiomyopathy (DCM) when their posture changes. Sixty-four children diagnosed with DCM were recruited as research group and 55 healthy children as control group. T-wave amplitude and QT interval in ECG were recorded, and their differences between supine and orthostatic ECG were compared in both groups. Subsequently, the children with DCM were followed up and the differences before and after treatment compared. ① Comparisons in differences: Differences of T-wave amplitude in lead II and III, aVF, and V and differences of QT interval in lead II, aVL, aVF, and V were lower in the research group than in the control group. ② Logistic regression analysis and diagnostic test evaluation: The differences of T-wave amplitude in lead III and QT interval in lead aVL may have predictive value for DCM diagnosis. When their values were 0.00 mV and 30 ms, respectively, the sensitivity and specificity of the combined index were 37.5 and 83.6%. ③ Follow-up: In the response group, the T-wave amplitude difference in lead aVR increased and the difference of QT interval in lead V decreased after treatment. In the non-response group, there was no difference before and after treatment. When the combined index of the differences of T-wave amplitude difference in lead aVR and QT interval difference in lead V, respectively, were -0.05 mV and 5 ms, the sensitivity and specificity of estimating the prognosis of DCM were 44.4 and 83.3%. The differences of T-wave amplitude and QT interval may have a certain value to estimate DCM diagnosis and prognosis.
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http://dx.doi.org/10.3389/fped.2021.680923DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8290918PMC
July 2021

Detecting the formation of human c-KIT oncogene promoter G-Quadruplex by Taylor dispersion analysis.

Talanta 2021 Oct 19;233:122533. Epub 2021 May 19.

Department of Chemistry, University of British Columbia, Vancouver, BC, V6T 1Z4, Canada. Electronic address:

The formation of G-quadruplex (G4) structures in oncogenic G-rich promoter regions are implicated in their biological functions, especially the inhibition of transcription. The binding of cations is thought to contribute to the stabilization of the G4 formation and competition against the duplex formation in the genomic sequence. Furthermore, it might affect the recognition of DNA-binding proteins. Therefore, measuring the interaction between G4 DNA and cations in a free solution environment is critical for evaluating G4 DNA biological functions. However, how binding to cations (K and NH) affects the folding equilibrium of the G4 structure remains unclear. In this work, a Taylor dispersion analysis (TDA) method using a capillary electrophoresis (CE) instrument was established for the quantitative characterization of the cation-dependent G4 formation in the human c-KIT oncogene promoter region, as well as diffusivities and hydrodynamic radii of DNA variations before and after folding. Our results showed that both K and NH can induce the random-coiled c-KIT DNA to unfold and form a more unstretched intermediate state and then fold into tightly structured G4s with smaller size. The G4 size induced by NH was smaller than that induced by K ions, though these two cations induced the c-KIT G4 DNA formation with similar binding constants (order of magnitude around 10 M). The TDA method can be widely used for rapid structural analyses of trace amounts of DNA mixtures, which effectively differentiate DNA variations or DNA-ligand complex conformations.
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http://dx.doi.org/10.1016/j.talanta.2021.122533DOI Listing
October 2021

[Determination of the binding of natural products to the human c-myb oncogene promoter G-quadruplex DNA by capillary electrophoresis and electrospray ionization mass spectrometry].

Se Pu 2020 Sep;38(9):1069-1077

Department of Chemistry, University of British Columbia, Vancouver, BC V6T 1Z1, Canada.

The relationship between a drug and its target directly affects its pharmacology and efficacy. Drug-target binding ability and binding stoichiometry are essential characterization data in pharmaceutical research. The c-myb proto-oncogene encodes a crucial transcription factor that is involved in proliferation, differentiation, and maturation during hematopoiesis. Recent studies have found that the human oncogene c-myb is overexpressed in cancer tissues such as colorectal cancer. C-myb has become a potential therapeutic target for colorectal cancer, leukemia, and other cancers. A guanine (G)-rich DNA sequence located in the promoter region of c-myb can be spontaneously folded to form an intra-molecular G-quadruplex (G4) with cationic induction. The specific recognition of small molecules can stabilize this G4 folding, thus regulating the transcription and expression of c-myb. In this study, pressure assisted capillary electrophoresis frontier analysis (PACE-FA) combined with electrospray ionization mass spectrometry (ESI-MS) was used to investigate the interactions between the human c-myb promoter G4 and natural product molecules. In PACE-FA, an external pressure (no more than 13.8 kPa) in the same direction of the migration of the analyte was used in capillary electrophoresis frontier analysis (CE-FA), which greatly sped up the analysis while maintaining the accuracy of the results. Meanwhile, the combination of PACE-FA and ESI-MS could rapidly determine the affinity and stoichiometric relationship between binding molecules and targets. First, the intramolecular parallel-stranded G4 formation of the c-myb promoter sequence in the presence of cations (K, NH) was investigated by circular dichroism (CD) and ESI-MS. Then, ESI-MS was used to rapidly screen the natural products for candidate molecules that bound the target G4 DNA. The binding interactions were measured by mixing the c-myb DNA with each natural product separately in a 1:4 molar ratio, and then directly infusing these mixtures into the ESI-MS system. From the ESI-MS spectra, values calculated from the relative intensities of DNA and its complex ions were used to probe the binding affinities of the natural products. This parameter denotes the relative binding affinity for a small molecule with the G4 DNA. Three natural products were identified through the screen, and their binding affinities to G4 DNA were ranked as follows:pseudolaric acid B > scopolamine butylbromide > nuciferine. Considering that both specific and non-specific binding existed in the solution phase, a free solution method using PACE-FA was developed to further test the binding ability of these products to the c-myb promoter G4 DNA. In the PACE-FA experiments, the pre-equilibrated mixture of the c-myb G4 DNA and the selected ligand was injected into the capillary prior to separation. Upon applying voltage and an external pressure (6.9 kPa) to the capillary, different species of analytes in the sample migrated at their own rates due to their different sizes and charges. The results showed that scopolamine butylbromide could bind specifically to target G4 DNA with 1:1 stoichiometry and a binding constant of 1.18×10 L/mol. Nuciferine's binding to G4 DNA showed a linear increasing trend due to nonspecific binding; thus nuciferine is a nonspecific binder. Although pseudolaric acid B showed high affinity for the c-myb G4 DNA and 1:1 and 1:2 G4-bound complex ions were observed in ESI-MS measurements, the PACE-FA results indicated that pseudolaric acid B did not bind to target G4 DNA in free solution. Therefore, scopolamine butylbromide could be the best candidate to regulate the transcription of the c-myb oncogene, and is expected to be a precursor for anticancer drugs. In this work, PACE-FA has allowed for significant improvements to the conventional CE-FA technique. This combination of PACE-FA and ESI-MS not only reduced the time needed for binding analysis, but also improved the accuracy and specificity of the affinity analysis compared to conventional binding approaches. Furthermore, this combination could be used to screen other targeted drug candidates and to evaluate their interaction mechanisms.
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http://dx.doi.org/10.3724/SP.J.1123.2020.03001DOI Listing
September 2020

Construction of a Three-Dimensional BaTiO Network for Enhanced Permittivity and Energy Storage of PVDF Composites.

Materials (Basel) 2021 Jun 27;14(13). Epub 2021 Jun 27.

School of Chemistry and Biological Engineering, University of Science and Technology Beijing, Beijing 100083, China.

Three-dimensional BaTiO (3D BT)/polyvinylidene fluoride (PVDF) composite dielectrics were fabricated by inversely introducing PVDF solution into a continuous 3D BT network, which was simply constructed via the sol-gel method using a cleanroom wiper as a template. The effect of the 3D BT microstructure and content on the dielectric and energy storage properties of the composites were explored. The results showed that 3D BT with a well-connected continuous network and moderate grain sizes could be easily obtained by calcining a barium source containing a wiper template at 1100 °C for 3 h. The as-fabricated 3D BT/PVDF composites with 21.1 wt% content of 3D BT (3DBT-2) exhibited the best comprehensive dielectric and energy storage performances. An enhanced dielectric constant of 25.3 at 100 Hz, which was 2.8 times higher than that of pure PVDF and 1.4 times superior to the conventional nano-BT/PVDF 25 wt% system, was achieved in addition with a low dielectric loss of 0.057 and a moderate dielectric breakdown strength of 73.8 kV·mm. In addition, the composite of 3DBT-2 exhibited the highest discharge energy density of 1.6 × 10 J·cm under 3 kV·mm, which was nearly 4.5 times higher than that of neat PVDF.
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http://dx.doi.org/10.3390/ma14133585DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8269637PMC
June 2021

Evaluation of the AGCU Expressmarker 30 Kit composed of 31 loci for forensic application.

Forensic Sci Int 2021 Jul 19;324:110849. Epub 2021 May 19.

Department of Forensic Genetics, West China School of Basic Medical Sciences & Forensic Medicine, Sichuan University, Chengdu 610041, China. Electronic address:

With the widespread use of STR in identification of individuals, paternity testing, as well as population genetics, many commercially robust and validated STR multiplex kits were developed. The AGCU Expressmarker 30 Kit is a new autosomal STR system that contains 29 autosomal STR loci (D3S1358, vWA, D1S1656, CSF1PO, D8S1132, D19S253, D3S3045, D8S1179, D21S11, D16S539, TPOX, D6S477, Penta D, D2S441, D5S818, TH01, FGA, D15S659, D22S1045, D19S433, D13S317, D7S820, D6S1043, D10S1435, D10S1248, D2S1338, D18S51, D12S391, and Penta E), one insertion/deletion polymorphic marker on the Y chromosome (Y indel), and the amelogenin locus. A series of validation studies were performed in this context according to the guidelines of "Validation Guidelines for Forensic DNA Analysis Methods". The sensitivity study showed that a full profile was observed with template DNA as low as 40 pg. In the stability study, all STR profiles were obtained at concentrations of humic acid up to 800 ng/μL, hematin up to 250 μM, and tannic acid up to 200 ng/μL. The mixture study demonstrated that all of the minor alleles could be called at ratios from 1:1-29:1 when the total DNA was 2 ng. In the population study, the total discrimination power for three population (Sichuan-Han, Gansu-Hui, and Guangxi-Zhuang) were above 0.9999999999999999999999999999999992, 0.999999999999999999999999999999998 and 0.999999999999999999999999999999994 as well as the cumulative probability of paternity exclusion were 0.999999999999953, 0.999999999999178, and 0.999999999999611 respectively. These results demonstrated that the AGCU Expressmarker 30 Kit is a useful tool for analyzing both forensic casework and database samples.
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http://dx.doi.org/10.1016/j.forsciint.2021.110849DOI Listing
July 2021

Quantitative characterization of human oncogene promoter G-quadruplex DNA-ligand interactions using a combination of mass spectrometry and capillary electrophoresis.

Electrophoresis 2021 08 28;42(14-15):1450-1460. Epub 2021 May 28.

Department of Chemistry, University of British Columbia, Vancouver, British Columbia, Canada.

Human c-KIT oncogene is known to regulate cell growth and proliferation, and thus, acts as a probable target in the treatment of gastrointestinal tumors (GIST). To identify small molecule ligands which can specifically bind with the G-quadruplex (G4) in the c-KIT promoter region as potential antitumor agents, we propose the combination of electrospray ionization-mass spectrometry (ESI-MS), capillary electrophoresis frontal analysis (CE-FA), and Taylor dispersion analysis (TDA) to accurately investigate the G4/ligands binding properties. First, ESI-MS was used for initial screening of natural products (NPs). CE-FA was then used to calculate specific binding constants and the stoichiometry of the native state binding pair in solution. Next, TDA, a micro-capillary flow technique was used to examine the effect of the ligand binding on the diffusivity and particle size of the c-KIT G4. Two of the screened NPs, scopolamine butylbromide (L1) and isorhamnetin-3-O-neohesperidoside (L3), were found to specifically bind to the c-KIT G4 with binding constants of around 10 M and 1:1 stoichiometry in a free solution. TDA data showed that ligand binding (both L1 and L3) induced the c-KIT strands to fold into a tightly structured G4 with a decreased hydrodynamic radius. These ligands have the potential to be drug candidates for the regulation of c-KIT gene transcription by stabilizing the G4 structure. This methodology not only increased the speed of analysis but also improved its accuracy and specificity compared with the conventional binding approaches.
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http://dx.doi.org/10.1002/elps.202100077DOI Listing
August 2021

Association between gonadal hormones and osteoporosis in schizophrenia patients undergoing risperidone monotherapy: a cross-sectional study.

PeerJ 2021 27;9:e11332. Epub 2021 Apr 27.

Department of Psychiatry, Suzhou Guangji Hospital, the Affiliated Guangji Hospital of Soochow University, Suzhou, Jiangsu, China.

Objective: Patients with schizophrenia are at increased risk of osteoporosis. This study first determined the osteoporosis rate in patients with schizophrenia and then then explored the association between serum gonadal hormone levels and osteoporosis among these patients.

Methods: A total of 250 patients with schizophrenia and 288 healthy controls were recruited. Osteoporosis was defined by decreased bone mineral density (BMD) of the calcaneus. Serum fasting levels of gonadal hormones (prolactin, estradiol, testosterone, progesterone, follicle-stimulating hormone, luteinizing hormone) were determined. The relationship between osteoporosis and hormone levels was statistically analyzed by binary logistic regression analysis.

Results: Our results showed that patients with schizophrenia had a markedly higher rate of osteoporosis (24.4% vs. 10.1%) than healthy controls ( < 0.001). Patients with osteoporosis were older, had a longer disease course, and had a lower body mass index (BMI) than patients without osteoporosis (all  < 0.05). Regarding gonadal hormones, we found significantly higher prolactin, but lower estradiol, levels in patients with osteoporosis than in those without osteoporosis (both  < 0.05). The regression analysis revealed that PRL (OR = 1.1, 95% CI [1.08-1.15],  < 0.001) and E2 level (OR = 0.9, 95%CI [0.96-0.99],  = 0.011) were significantly associated with osteoporosis in patients with schizophrenia.

Conclusion: Our results indicate that patients with schizophrenia who are being treated with risperidone have a high rate of osteoporosis. Increased prolactin and reduced estradiol levels are significantly associated with osteoporosis.
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http://dx.doi.org/10.7717/peerj.11332DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8086585PMC
April 2021

A qualitative study of home health care experiences among Chinese homebound adults.

BMC Geriatr 2021 05 13;21(1):309. Epub 2021 May 13.

Centre for Health Management and Policy Research, School of Public Health, Cheeloo College of Medicine, Shandong University, 44 Wenhuaxi Rd, Jinan, 250012, Shandong, China.

Background: Home health care services (HHC) are emerging in China to meet increased healthcare needs among the homebound population, but there is a lack of research examining the efficiency and effectiveness of this new care model. This study aimed to investigate care recipients' experiences with HHC and areas for improvement in China.

Methods: This research was a qualitative study based on semi-structured interviews. Qualitative data were collected from homebound adults living in Jinan, Zhangqiu, and Shanghai, China. A sample of 17 homebound participants aged 45 or older (mean age = 76) who have received home-based health care were recruited. Conceptual content analysis and Colaizzi's method was used to generate qualitative codes and identify themes.

Results: The evaluations of participants' experiences with HHC yielded both positive and negative aspects. Positive experiences included: 1) the healthcare delivery method was convenient for homebound older adults; 2) health problems could be detected in a timely manner because clinicians visited regularly; 3) home care providers had better bedside manners and technical skills than did hospital-based providers; 4) medical insurance typically covered the cost of home care services. Areas that could potentially be improved included: 1) the scope of HHC services was too limited to meet all the needs of homebound older adults; 2) the visit time was too short; 3) healthcare providers' technical skills varied greatly.

Conclusions: Findings from this study suggested that the HHC model benefited Chinese older adults-primarily homebound adults-in terms of convenience and affordability. There are opportunities to expand the scope of home health care services and improve the quality of care. Policymakers should consider providing more resources and incentives to enhance HHC in China. Educational programs may be created to train more HHC providers and improve their technical skills.
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http://dx.doi.org/10.1186/s12877-021-02258-yDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8117649PMC
May 2021

Influence of Core Competence on Voice Behavior of Clinical Nurses: A Multicenter Cross-Sectional Study.

Psychol Res Behav Manag 2021 29;14:501-510. Epub 2021 Apr 29.

School of Nursing, Hebei University, Baoding, Hebei, 07100, People's Republic of China.

Background: Voice behavior, referred to as a positive guarantee for organizational development, is influenced by several kinds of individual, collective and organizational features. However, the impact of individual competence on voice behavior is unclear. The aim of the present study was to investigate the status quo of core competence and voice behavior of clinical nurses and explore the impact of core competence on nurses' voice behavior.

Methods: A multicenter cross-sectional survey. A total of 1717 nurses were recruited from nine tertiary and secondary hospitals between March and June 2019. An online questionnaire, including socio-demographic variables, employee voice behavior scale and competence inventory for registered nurses, was used to investigate prohibitive and promotive voice behavior and core competence of clinical nurses. Pearson correlation and hierarchical multiple regression were performed in the data analysis.

Results: The mean score for prohibitive and promotive voice behavior of nurses were 3.46 (SD 0.77) and 3.46 (SD 0.88), respectively. The mean score for core competence was 2.46 (SD 0.77). Critical thinking/research aptitude was the most important predictor for both prohibitive and promotive voice behavior (each < 0.05), but its influence on promotive voice behavior was greater ( < 0.05). Leadership was another significant predictor for prohibitive voice behavior ( < 0.05). Legal/ethical practice, teaching-coaching, professional development and shift work were other predictors for promotive voice behavior (each < 0.05).

Conclusion: Clinical nurses experience modest levels of prohibitive and promotive voice behavior and their core competence is moderate. Core competence, especially critical thinking/research aptitude, impacts significantly on voice behavior of clinical nurses. Cultivating nurses' core competence could positively increase their voice behavior for organizational development.
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http://dx.doi.org/10.2147/PRBM.S309565DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8092618PMC
April 2021

A General Nanocoating Method via Photoinduced Self-Initiation.

Langmuir 2021 May 27;37(18):5548-5553. Epub 2021 Apr 27.

Key Laboratory of Artificial Micro- and Nano-structures of Ministry of Education of China, School of Physics and Technology, Wuhan University, Wuhan 430072 China.

Hybrid core-shell nanoparticles play a very significant role in many applications. Here, we report a light-induced oligomer coating on nanoparticles via Norrish type I reaction. The radical species generated via UV irradiation can chemically initiate the photoinitiators, which are then polymerized and deposited on inorganic nanoparticles via heterogeneous nucleation, forming a soft oligomer coating smaller than 40 nm. This coating method is versatile and potentially applicable to many different types of inorganic cores and their assemblies, making it a very useful technique for "freezing" nanoassemblies in solution. Moreover, these oligomer coatings containing radical species can also initiate surface polymerization of both styrenic and acrylic monomers with certain functionalities for different applications such as self-assembly, plasmon tuning, and pH sensing (3.5-4.5).
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http://dx.doi.org/10.1021/acs.langmuir.1c00303DOI Listing
May 2021

Multi-omics analysis to visualize the dynamic roles of defense genes in the response of tea plants to gray blight.

Plant J 2021 05 26;106(3):862-875. Epub 2021 Mar 26.

State Key Laboratory of Tea Plant Biology and Utilization, Anhui Agricultural University, Hefei, Anhui, China.

Gray blight (GB) is one of the most destructive diseases of tea plants, causing considerable damage and productivity losses; however, the dynamic roles of defense genes during pathogen infection remain largely unclear. To explore the numerous molecular interactions associated with GB stress in tea plants, we employed transcriptome, sRNAome and degradome sequencing from 1 to 13 days post-inoculation (dpi) at 3-day intervals. The transcriptomics results showed that differentially expressed genes (DEGs) related to flavonoid synthesis, such as chalcone synthase (CHS) and phenylalanine ammonia-lyase (PAL), were particularly induced at 4 dpi. Consistent with this, the contents of catechins (especially gallocatechin), which are the dominant flavonoids in tea plants, also increased in the leaves of tea plants infected with GB. Combined analysis of the sRNAome and degradome revealed that microRNAs could mediate tea plant immunity by regulating DEG expression at the post-transcriptional level. Co-expression network analysis demonstrated that miR530b-ethylene responsive factor 96 (ERF96) and miRn211-thaumatin-like protein (TLP) play crucial roles in the response to GB. Accordingly, gene-specific antisense oligonucleotide assays suggested that suppressing ERF96 decreased the levels of reactive oxygen species (ROS), whereas suppressing TLP increased the levels of ROS. Furthermore, ERF96 was induced, but TLP was suppressed, in susceptible tea cultivars. Our results collectively demonstrate that ERF96 is a negative regulator and TLP is a positive regulator in the response of tea plants to GB. Taken together, our comprehensive integrated analysis reveals a dynamic regulatory network linked to GB stress in tea plants and provides candidate genes for improvement of tea plants.
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http://dx.doi.org/10.1111/tpj.15203DOI Listing
May 2021

Therapeutic potential of coumestan Pks13 inhibitors for tuberculosis.

Antimicrob Agents Chemother 2021 Feb 8. Epub 2021 Feb 8.

Center for Tuberculosis Research, Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, MD, USA

Polyketide synthase 13 (Pks13) is an important enzyme found in () that condenses two fatty acyl chains to produce α-alkyl β-ketoesters, which in turn serve as the precursors for the synthesis of mycolic acids that are essential building blocks for maintaining the cell wall integrity of Coumestan derivatives have recently been identified in our group as a new chemotype that exert their antitubercular effects via targeting of Pks13. These compounds were active on both drug-susceptible and drug-resistant strains of as well as showing low cytotoxicity to healthy cells and a promising selectivity profile. No cross-resistance was found between the coumestan derivatives and first-line TB drugs. Here we report that treatment of bacilli with 15 times the MIC of compound , an optimized lead coumestan compound, resulted in a colony forming unit (CFU) reduction from 6.0 log units to below the limit of detection (1.0 log units) per mL culture, demonstrating a bactericidal mechanism of action. Single dose (10 mg/kg) pharmacokinetic studies revealed favorable parameters with a relative bioavailability of 19.4%. In a mouse infection and chemotherapy model, treatment with showed dose-dependent mono-therapeutic activity, whereas treatment with in combination with rifampin showed clear synergistic effects. Together these data suggest that coumestan derivatives are promising agents for further TB drug development.
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http://dx.doi.org/10.1128/AAC.02190-20DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8092898PMC
February 2021

Ultrasound cycloplasty in Chinese glaucoma patients: Results of a 6-month prospective clinical study.

Ophthalmic Res 2021 Feb 4. Epub 2021 Feb 4.

Purpose: The aim of this study was to evaluate the short-term efficacy and safety of ultrasound cycloplasty (UCP) procedure in Chinese glaucoma patients.

Methods: As a single-centre, prospective, non-comparative study, 23 eyes of 23 patients suffering from glaucoma with uncontrolled intraocular pressure (IOP) ≥ 21 mmHg underwent a multi-dose UCP treatment with the activations of 6, 8 or10 sectors. Types of glaucoma include primary open-angle glaucoma (POAG) (7/23), primary angle closure glaucoma (PACG) (9/23) and secondary glaucoma (SG) (7/23). A complete ophthalmic examination including intraocular pressure (IOP) measurements was performed before UCP procedure and at 1 day, 1 month, 3months and 6 months after the procedure. An IOP reduction of ≥20% and IOP > 5 mmHg without increasing hypotensive medication at the follow-up visit was defined as therapeutic success. The post-operative complications were also recorded and compared to baseline for safety evaluation.

Results: The mean baseline IOP of 23 treated eyes was 37.2 ± 12.1 mmHg. The IOP reduction after UCP procedure were 23%, 49%, 33% and 34% at 1 day, 1 month, 3 months and 6 months, respectively. Thus, the corresponding overall therapeutic success rates reached 61% (14/23), 83% (19/23), 65% (15/23) and 61% (14/23), respectively. Baseline IOPs of 8 and 10 sectors group (37.0 ± 9.9 mmHg and 50.1 ± 12.2 mmHg) were significantly higher than that of 6 sectors group (30.1 ± 8.2 mmHg). Therapeutic success rates of 6, 8 and 10 sectors groups reached 44% (4/9), 56% (5/9) and 100% (5/5), respectively. There were the highest percentage of IOP reduction (50% and 41%) and therapeutic success rate (6/7; 86% and 7/9; 78%) in SG group and PACG groups respectively. In addition, pre-operative ocular pain symptoms of four patients were all disappeared within one week after UCP. No serious intra-operative or post-operative complications occurred.

Conclusion: UCP procedure is an effective and well-tolerated treatment to reduce IOP in Chinese glaucoma patients, which offered a novel alternative for glaucoma treatment.
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http://dx.doi.org/10.1159/000515013DOI Listing
February 2021

Rapid and simultaneous detection of Japanese encephalitis virus by real-time nucleic acid sequence-based amplification.

Microb Pathog 2021 Jan 2;150:104724. Epub 2021 Jan 2.

Key Laboratory of Prevention and Control Agents for Animal Bacteriosis (Ministry of Agriculture), Institute of Animal Husbandry and Veterinary, Hubei Academy of Agricultural Sciences, Wuhan, 430064, China. Electronic address:

Japaneses encephalitis (JE) is most common zoonoses caused by Japanese encephalitis virus (JEV) with a high mortality and disability rate. To take timely preventive and control measures, early and rapid detection of JE RNA is necessary. But due to characteristic brief and low viraemia, JE RNA detection remains challenging. In this study, a real-time nucleic acid sequence-based amplification (RT-NASBA) was developed for rapid and simultaneous detection of JEV. Four pairs of primer were designed using a multiple genome alignment of all JEV strains from GenBank. NASBA assay established and optimal reaction conditions were confirmed by using primers and probe on ns1 gene of JEV. The specificity and sensitivity of the assay were compared with RT-PCR by using serial RNA and virus cultivation dilutions. The results showed that JEV RT-NASBA assay was established, and robust signals could be observed in 10 min with high specificity. The limit of dectetion of RT-NASBA was 6 copies per reaction. The assay was thus 100 to 1, 000 times more sensitive than RT-PCR. The cross-reaction was performed with other porcine pathogens, and negative amplification results indicated the high specificity of this method. The novel JEV RT-NASBA assay could be used as an efficient molecular biology tool to diagnose JEV, which would facilitate the surveillance of reproductive failure disease in swine and would be beneficial for public health security.
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http://dx.doi.org/10.1016/j.micpath.2020.104724DOI Listing
January 2021

ATF-3 expression inhibits melanoma growth by downregulating ERK and AKT pathways.

Lab Invest 2021 05 9;101(5):636-647. Epub 2020 Dec 9.

Department of Pathology, Program in Dermatopathology, Brigham and Women's Hospital, Boston, MA, USA.

Activating transcription factor 3 (ATF-3), a cyclic AMP-dependent transcription factor, has been shown to play a regulatory role in melanoma, although its function during tumor progression remains unclear. Here, we demonstrate that ATF-3 exhibits tumor suppressive function in melanoma. Specifically, ATF-3 nuclear expression was significantly diminished with melanoma progression from nevi to primary to metastatic patient melanomas, correlating low expression with poor prognosis. Significantly low expression of ATF-3 was also found in cultured human metastatic melanoma cell lines. Importantly, overexpression of ATF-3 in metastatic melanoma cell lines significantly inhibited cell growth, migration, and invasion in vitro; as well as abrogated tumor growth in a human melanoma xenograft mouse model in vivo. RNA sequencing analysis revealed downregulation of ERK and AKT pathways and upregulation in apoptotic-related genes in ATF-3 overexpressed melanoma cell lines, which was further validated by Western-blot analysis. In summary, this study demonstrated that diminished ATF-3 expression is associated with melanoma virulence and thus provides a potential target for novel therapies and prognostic biomarker applications.
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http://dx.doi.org/10.1038/s41374-020-00516-yDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8091967PMC
May 2021

Cloning of wheat keto-acyl thiolase 2B reveals a role of jasmonic acid in grain weight determination.

Nat Commun 2020 12 8;11(1):6266. Epub 2020 Dec 8.

State Key Laboratory of Genetic Engineering, MOE Key Laboratory for Biodiversity Science and Ecological Engineering, MOE Engineering Research Center of Gene Technology, Institute of Plant Biology, School of Life Sciences, Fudan University, Shanghai, 200438, China.

Grain weight (GW) is one of the component traits of wheat yield. Existing reports have shown that multiple phytohormones are involved in the regulation of GW in different crops. However, the potential role of jasmonic acid (JA) remains unclear. Here, we report that triticale grain weight 1 (tgw1) mutant, with marked reductions in both GW and JA content, is caused by a premature stop mutation in keto-acyl thiolase 2B (KAT-2B) involved in β-oxidation during JA synthesis. KAT-2B overexpression increases GW in wild type and boosts yield. Additionally, KAT-2B compliments the grain defect in tgw1 and rescues the lethal phenotype of the Arabidopsis kat2 mutant in a sucrose-free medium. Despite the suppression of JA synthesis in tgw1 mutant, ABA synthesis is upregulated, which is accompanied by enhanced expression of SAG3 and reduction of chlorophyll content in leaves. Together, these results demonstrate a role of the JA synthetic gene KAT-2B in controlling GW and its potential application value for wheat improvement.
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http://dx.doi.org/10.1038/s41467-020-20133-zDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7722888PMC
December 2020

Acvr1 deletion in osteoblasts impaired mandibular bone mass through compromised osteoblast differentiation and enhanced sRANKL-induced osteoclastogenesis.

J Cell Physiol 2021 06 29;236(6):4580-4591. Epub 2020 Nov 29.

Department of Oral Pathology, Hospital of Stomatology, Jilin University, Changchun, China.

Bone morphogenetic protein (BMP) signaling is well known in bone homeostasis. However, the physiological effects of BMP signaling on mandibles are largely unknown, as the mandible has distinct functions and characteristics from other bones. In this study, we investigated the roles of BMP signaling in bone homeostasis of the mandibles by deleting BMP type I receptor Acvr1 in osteoblast lineage cells with Osterix-Cre. We found mandibular bone loss in conditional knockout mice at the ages of postnatal day 21 and 42 in an age-dependent manner. The decreased bone mass was related to compromised osteoblast differentiation together with enhanced osteoclastogenesis, which was secondary to the changes in osteoblasts in vivo. In vitro study revealed that deletion of Acvr1 in the mandibular bone marrow stromal cells (BMSCs) significantly compromised osteoblast differentiation. When wild type bone marrow macrophages were cocultured with BMSCs lacking Acvr1 both directly and indirectly, both proliferation and differentiation of osteoclasts were induced as evidenced by an increase of multinucleated cells, compared with cocultured with control BMSCs. Furthermore, we demonstrated that the increased osteoclastogenesis in vitro was at least partially due to the secretion of soluble receptor activator of nuclear factor-κB ligand (sRANKL), which is probably the reason for the mandibular bone loss in vivo. Overall, our results proposed that ACVR1 played essential roles in maintaining mandibular bone homeostasis through osteoblast differentiation and osteoblast-osteoclast communication via sRANKL.
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http://dx.doi.org/10.1002/jcp.30183DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8048423PMC
June 2021

Discovery of γ-Lactam Alkaloid Derivatives as Potential Fungicidal Agents Targeting Steroid Biosynthesis.

J Agric Food Chem 2020 Dec 22;68(49):14438-14451. Epub 2020 Nov 22.

National Biopesticide Engineering Research Centre, Hubei Biopesticide Engineering Research Centre, Hubei Academy of Agricultural Science, Wuhan 430064, China.

Biological control of plant pathogens is considered as one of the green and effective technologies using beneficial microorganisms or microbial secondary metabolites against plant diseases, and so microbial natural products have played important roles in the research and development of new and green agrochemicals. To explore the potential applications for natural γ-lactam alkaloids and their derivatives, 26 γ-lactams that have flexible substituent patterns were synthesized and characterized, and their antifungal activities against eight kinds of plant pathogens belonging to oomycetes, basidiomycetes, and deuteromycetes were fully evaluated. In addition, the high potential compounds were further tested using an assay against blight of pepper to verify a practical application for controlling oomycete diseases. The potential modes of action for compound against were also investigated using microscopic technology (optical microscopy, scanning electron microscopy, and transmission electron microscopy) and label-free quantitative proteomics analysis. The results demonstrated that compound may be a potential novel fungicidal agent against oomycete diseases (EC = 4.9748 μg·mL for and EC = 5.1602 μg·mL for ) that can act on steroid biosynthesis, which can provide a certain theoretical basis for the development of natural lactam derivatives as potential antifungal agents.
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http://dx.doi.org/10.1021/acs.jafc.0c05823DOI Listing
December 2020

Calcified Aortic Valve Disease in Patients With Familial Hypercholesterolemia.

J Cardiovasc Pharmacol 2020 11;76(5):506-513

School of Medicine, Ningbo University, Ningbo, Zhejiang, China; and.

Familial hypercholesterolemia (FH) is a rare autosomal gene deficiency disease with increased low-density lipoprotein cholesterol, xanthoma, and premature coronary heart disease. Calcified aortic valve disease (CAVD) is prevalent in FH patients, resulting in adverse events and heavy health care burden. Aortic valve calcification is currently considered an active biological process, which shares several common risk factors with atherosclerosis, including aging, hypertension, dyslipidemia, and so on. Unfortunately, the pathogenesis and therapy of CAVD in FH are still controversial. There is no pharmacological intervention recommended to delay the development of CAVD in FH, and the only effective treatment for severe CAVD is aortic valve replacement. In this review, we summarize the detailed description of the pathophysiology, molecular mechanism, risk factors, and treatment of CAVD in FH patients.
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http://dx.doi.org/10.1097/FJC.0000000000000890DOI Listing
November 2020

Developmental validation of the Yfiler Platinum PCR Amplification Kit for forensic genetic caseworks and databases.

Electrophoresis 2021 01 22;42(1-2):126-133. Epub 2020 Nov 22.

Institute of Forensic Medicine, West China School of Basic Sciences and Forensic Medicine, Sichuan University, Chengdu, P. R. China.

Y chromosome kits are successfully applied in cases where human biological material exists. With the development of genotyping ability, more Y chromosomal markers are needed for finer identification of male individuals and lineages. In this study, a developmental validation of a newly emerged Y chromosome kit that combines two different kinds of markers: 38 Y-STRs and 3 Y-indels are conducted. The results show that this kit has high sensitivity when there is a small amount of DNA (125 pg), more than one male (minor:major = 1:7), or a mixture of males and females (male:female = 125pg:1875pg), inhibited substances (800 μM hematin and more than 1600 ng/μL humic acid). The kit exhibits high precision level with a standard deviation of allele size no more than 0.14 nt. Locus DYS481 shows the largest stutter rate, with three stutters per true allele. Population samples are well identified (MP of 0.001106), and mutations can be observed in father-son pairs (46 mutations in 70 pairs, 10 in locus DYS627). Out of all the population samples, 13.2% belong to haplogroup M117-O2a2b1a1, with their ethnic group being Han Chinese. The results show that this kit can improve the performance of identifying male individuals, obtaining more unique haplotypes (increasing from 894 to 918 of 1000 male samples) and higher discrimination capacity (increasing from 0.942 to 0.955) in this study compared to previous widely used Yfiler Plus kit. Besides, it gives information about their paternal lineages in forensic genetic casework and genealogical database construction.
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http://dx.doi.org/10.1002/elps.202000187DOI Listing
January 2021

Substantial Differences in Turnover Intention Between Direct Care Workers in Chinese Hospitals and Long-Term Care Facilities.

J Am Med Dir Assoc 2021 03 21;22(3):696-700.e1. Epub 2020 Oct 21.

Shandong University, Cheeloo College of Medicine, School of Public Health, Centre for Health Management and Policy Research (NHC Key Lab of Health Economics and Policy Research), Jinan, Shandong, China; Home Centered Care Institute, Schaumburg, IL, USA; School of Medicine, University of Virginia, Charlottesville, VA, USA. Electronic address:

Objectives: High retention rates among direct care workers (DCWs) affect the quality of aged care. However, limited research has explored factors associated with retention in China's aged care industry. This study compared turnover intention among DCWs in Chinese hospitals and long-term care facilities (LTCFs).

Design: A cross-sectional survey with the stratified-random sampling method.

Setting And Participants: A total of 370 DCWs from 7 hospitals and 311 DCWs from 7 LTCFs located in Fujian Province, China, were surveyed.

Measures: Turnover intention, contractual status, income, sociodemographic characteristics, and other work-related factors were self-reported by surveyed DCWs.

Results: Approximately 80% of DCWs in hospitals had employment contracts, whereas only 9% of DCWs were contractual in LTCFs. DCWs from hospitals reported lower turnover intention (20.5% vs. 37.0%) than did DCWs from LTCFs. Contracted DCWs from hospitals had the lowest turnover intention (14.8%). Contracted and noncontracted DCWs in LTCFs were more likely to report turnover intention than hospital contracted DCWs (odds ratio [OR] 3.68, P = .008 and OR = 3.17, P < .001, respectively).

Conclusions And Implications: Contractual status and facility type were jointly associated with turnover intention of DCWs in China. DCWs in LTCFs were likely to report much higher turnover intention than contracted DCWs in hospitals. The expanded use of employment contracts may lower turnover intention in long-term care DCWs by improving their job stability and security. The labor market of DCWs needs to be regulated by the Chinese Labor Law.
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http://dx.doi.org/10.1016/j.jamda.2020.09.006DOI Listing
March 2021

Simultaneous measurement of the BOD concentration and temperature based on a tapered microfiber for water pollution monitoring.

Appl Opt 2020 Aug;59(24):7364-7370

An optical sensor that simultaneously measures the concentration of the biochemical oxygen demand (BOD) and temperature in water based on a tapered microfiber is proposed for environmental monitoring. The sensor is characterized by a strong evanescent field, which is more sensitive to liquids with a low refractive index and a low transmission loss. The results show that as the BOD concentration increases, the interference spectrum shifts toward longer wavelengths, the spectral loss decreases, and the sensitivities of the BOD are 12.17 nm/mg/mL and -2.387// in the range of 0.25-1 mg/mL, which indicates the extent of the water pollution. The detection limit for the BOD concentration is as low as 0.0016 mg/mL. As the ambient temperature increases, the interference spectrum shifts toward shorter wavelengths, the spectral loss decreases, and the temperature sensitivities are -0.339/ and -0.031/ in the range of 30°C-60°C. The matrix method can be used to achieve the simultaneous measurement of the BOD concentration and environmental temperature because the spectral interference peaks have different responses to these two parameters. The sensor can not only be used for detecting water pollution in rivers, drinking water, and groundwater but can also be utilized for other types of environmental monitoring. This sensor has great potential to act as a basic sensing unit in fiber-optic sensor networks for multiparameter measurements and intelligent monitoring.
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http://dx.doi.org/10.1364/AO.396831DOI Listing
August 2020

A Removable Artificial Cell Wall for Withstanding Ciprofloxacin.

Macromol Biosci 2020 12 7;20(12):e2000185. Epub 2020 Sep 7.

School of Chemistry and Chemical Engineering, Harbin Institute of Technology, Harbin, 150001, P. R. China.

The pollution of antibiotics in aquaculture environment is increasingly serious, and excessive antibiotics will kill the probiotics in aquaculture feed. How to improve the viability of probiotics in the antibiotics-contaminated environment is of significance. In this study, a new strategy for protecting Saccharomyces cerevisiae cells in situ against antibiotics is constructed based on cell surface engineering technology by putting on wearable protective layers for cells. The protective layer is constructed around cellular surface via the self-assembly of coacervate microdroplets that consist of carboxymethyl chitosan and carboxyl dextran. Without affecting the cell viability, the protective layer can grasp ciprofloxacin and decrease the contact of ciprofloxacin to cells and consequently improve the survival rate of cells when exposing to ciprofloxacin. This work highlights a facile strategy to establish removable artificial cell wall by biodegradable polysaccharides for improving the productivity of probiotics in antibiotic environments.
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http://dx.doi.org/10.1002/mabi.202000185DOI Listing
December 2020

Global Quantitative Proteomics Studies Revealed Tissue-Preferential Expression and Phosphorylation of Regulatory Proteins in .

Int J Mol Sci 2020 Aug 25;21(17). Epub 2020 Aug 25.

State Key Laboratory of Genetic Engineering and Collaborative Innovation Center for Genetics and Development, Ministry of Education Key Laboratory of Biodiversity Sciences and Ecological Engineering and Institute of Biodiversity Sciences, Institute of Plant Biology, School of Life Sciences, Fudan University, Shanghai 200438, China.

Organogenesis in plants occurs across all stages of the life cycle. Although previous studies have identified many genes as important for either vegetative or reproductive development at the RNA level, global information on translational and post-translational levels remains limited. In this study, six stages/organs were analyzed using quantitative proteomics and phosphoproteomics, identifying 2187 non-redundant proteins and evidence for 1194 phosphoproteins. Compared to the expression observed in cauline leaves, the expression of 1445, 1644, and 1377 proteins showed greater than 1.5-fold alterations in stage 1-9 flowers, stage 10-12 flowers, and open flowers, respectively. Among these, 294 phosphoproteins with 472 phosphorylation sites were newly uncovered, including 275 phosphoproteins showing differential expression patterns, providing molecular markers and possible candidates for functional studies. Proteins encoded by genes preferentially expressed in anther (15), meiocyte (4), or pollen (15) were enriched in reproductive organs, and mutants of two anther-preferentially expressed proteins, and , showed obviously reduced male fertility with abnormally organized pollen exine. In addition, more phosphorylated proteins were identified in reproductive stages (1149) than in the vegetative organs (995). The floral organ-preferential phosphorylation of GRP17, CDC2/CDKA.1, and ATSK11 was confirmed with western blot analysis. Moreover, phosphorylation levels of CDPK6 and MAPK6 and their interacting proteins were elevated in reproductive tissues. Overall, our study yielded extensive data on protein expression and phosphorylation at six stages/organs and provides an important resource for future studies investigating the regulatory mechanisms governing plant development.
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http://dx.doi.org/10.3390/ijms21176116DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7503369PMC
August 2020

Prolonged treatment with Y-27632 promotes the senescence of primary human dermal fibroblasts by increasing the expression of IGFBP-5 and transforming them into a CAF-like phenotype.

Aging (Albany NY) 2020 08 25;12(16):16621-16646. Epub 2020 Aug 25.

Department of Tissue Engineering and Regeneration, School and Hospital of Stomatology, Cheeloo College of Medicine, Shandong University and Shandong Key Laboratory of Oral Tissue Regeneration and Shandong Engineering Laboratory for Dental Materials and Oral Tissue Regeneration, Jinan, China.

The Rho-kinases (ROCK) inhibitor Y-27632 has been shown to promote the growth of epidermal cells, however, its potential effects on human dermal fibroblasts (HDFs) need to be clarified. Here we show that prolonged treatment of HDFs with Y-27632 decreased their growth by inducing senescence, which was associated with induction of the senescence markers p16 and p21, and downmodulation of the ERK pathways. The senescent HDFs induced by Y-27632 acquired a cancer-associated-fibroblast (CAF)-like phenotype to promote squamous cell carcinoma (SCC) cell growth . Expression of a newly identified target of Y-27632 by RNA-seq, insulin growth factor binding protein 5 (IGFBP-5), was dramatically increased after 24 h of treatment with Y-27632. Adding recombinant IGFBP-5 protein to the culture medium produced similar phenotypes of HDFs as did treatment with Y-27632, and knockdown of IGFBP-5 blocked the Y-27632-induced senescence. Furthermore, Y-27632 induced the expression of an IGFBP-5 upstream gene, GATA4, and knockdown of GATA4 also reduced the Y-27632-induced senescence. In summary, these results demonstrate for the first time that Y-27632 promotes cellular senescence in primary HDFs by inducing the expression of IGFBP-5 and that prolonged treatment with Y-27632 potentially transforms primary HDFs into CAF-like cells.
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http://dx.doi.org/10.18632/aging.103910DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7485707PMC
August 2020

Y-27632 Enriches the Yield of Human Melanocytes from Adult Skin Tissues.

J Vis Exp 2020 07 8(161). Epub 2020 Jul 8.

Department of Tissue Engineering and Regeneration, School and Hospital of Stomatology, Cheeloo College of Medicine, Shandong University & Shandong Key Laboratory of Oral Tissue Regeneration & Shandong Engineering Laboratory for Dental Materials and Oral Tissue Regeneration;

The isolation and culture of primary melanocytes from skin tissues is very important for biological research and has been widely used for clinical applications. Isolating primary melanocytes from skin tissues by the conventional method usually takes about 3 to 4 weeks to passage sufficiently. More importantly, the tissues used are usually newborn foreskins and it is still a challenge to efficiently isolate primary melanocytes from adult tissues. We recently developed a new isolation method for melanocytes that adds Y-27632, a Rho kinase inhibitor, to the initial culture medium for 48 h. Compared with the conventional protocol, this new method dramatically increases the yield of melanocytes and shortens the time required to isolate melanocytes from foreskin tissues. We now describe this new method in more detail using adult epidermis to efficiently culture primary melanocytes. Importantly, we show that melanocytes obtained from adult tissues prepared by this new method can function normally. This new protocol will significantly benefit studies of pigmentation defects and melanomas using primary melanocytes prepared from easily accessed adult skin tissues.
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http://dx.doi.org/10.3791/61226DOI Listing
July 2020
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