Publications by authors named "Dan Chen"

1,223 Publications

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Biomaterials and advanced technologies for the evaluation and treatment of ovarian aging.

J Nanobiotechnology 2022 Aug 11;20(1):374. Epub 2022 Aug 11.

Department of Obstetrics and Gynecology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, Hubei, China.

Ovarian aging is characterized by a progressive decline in ovarian function. With the increase in life expectancy worldwide, ovarian aging has gradually become a key health problem among women. Over the years, various strategies have been developed to preserve fertility in women, while there are currently no clinical treatments to delay ovarian aging. Recently, advances in biomaterials and technologies, such as three-dimensional (3D) printing and microfluidics for the encapsulation of follicles and nanoparticles as delivery systems for drugs, have shown potential to be translational strategies for ovarian aging. This review introduces the research progress on the mechanisms underlying ovarian aging, and summarizes the current state of biomaterials in the evaluation and treatment of ovarian aging, including safety, potential applications, future directions and difficulties in translation.
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http://dx.doi.org/10.1186/s12951-022-01566-8DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC9367160PMC
August 2022

Circulating miR-448 acts as a potential diagnostic biomarker for multiple myeloma.

Hematology 2022 Dec;27(1):847-852

Department of Central Laboratory, The Affiliated Hospital of Chengde Medical College, Chengde, People's Republic of China.

Objective: The current diagnostic methods for multiple myeloma (MM) include bone marrow aspiration and biopsy, which are not only complicated but also invasive, causing great pain to patients. Micro ribonucleotides (miRNA) exist stably in circulating plasma and could be easily detected, making them promising specific diagnostic markers for MM.

Methods: The expression of plasma miR-448 in MM patients was detected by real-time quantitative PCR analysis. Spearman correlation was used to analyze correlations between miR-448 expression and various clinic pathological characteristics of MM patients. The ROC and the AUC (95% confidence interval) were exploited to evaluate the potential of miR-448 acting as a diagnostic marker for MM patients.

Results And Discussion: In this study, we found that the expression level of miR-448 in patients with MM was significantly higher than that in normal people and showed statistically differential expression in different stages of MM. Besides, we observed that the abundance of miR-448 in the plasma of newly diagnosed MM patients was positively correlated with the proportion of plasma cells (PC) in the bone marrow and the concentration of serum M protein (MP), respectively. In addition, ROC analysis demonstrated the sensitivity and specificity of the diagnostic value of miR-448 for MM are 92.90% and 87.50%, respectively.

Conclusion: Taken together, these results indicated that miR-448 may serve as a potential molecular diagnostic marker for MM.
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http://dx.doi.org/10.1080/16078454.2022.2106818DOI Listing
December 2022

Oxytocin and serotonin in the modulation of neural function: Neurobiological underpinnings of autism-related behavior.

Front Neurosci 2022 22;16:919890. Epub 2022 Jul 22.

Experimental Center, Shandong University of Traditional Chinese Medicine, Jinan, China.

Autism spectrum disorders (ASD) is a group of generalized neurodevelopmental disorders. Its main clinical features are social communication disorder and repetitive stereotyped behavioral interest. The abnormal structure and function of brain network is the basis of social dysfunction and stereotyped performance in patients with autism spectrum disorder. The number of patients diagnosed with ASD has increased year by year, but there is a lack of effective intervention and treatment. Oxytocin has been revealed to effectively improve social cognitive function and significantly improve the social information processing ability, empathy ability and social communication ability of ASD patients. The change of serotonin level also been reported affecting the development of brain and causes ASD-like behavioral abnormalities, such as anxiety, depression like behavior, stereotyped behavior. Present review will focus on the research progress of serotonin and oxytocin in the pathogenesis, brain circuit changes and treatment of autism. Revealing the regulatory effect and neural mechanism of serotonin and oxytocin on patients with ASD is not only conducive to a deeper comprehension of the pathogenesis of ASD, but also has vital clinical significance.
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http://dx.doi.org/10.3389/fnins.2022.919890DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC9354980PMC
July 2022

mTORC2 facilitates liver regeneration through the sphingolipid-induced PPAR-α-fatty acid oxidation.

Cell Mol Gastroenterol Hepatol 2022 Aug 2. Epub 2022 Aug 2.

Key Laboratory of Regenerative Medicine of Ministry of Education, Institute of Aging and Regenerative Medicine, Jinan University, Guangzhou, 510632, China. Electronic address:

Background & Aims: During liver regeneration following partial hepatectomy, the function and metabolic pathways governing transient lipid droplet accumulation in hepatocytes remain obscure. Mammalian target of rapamycin 2 (mTORC2) facilitates de novo synthesis of hepatic lipids. Under normal conditions and in tumorigenesis, decreased levels of triglyceride (TG) and fatty acids (FAs) are observed in the mTORC2-deficient liver. However, during liver regeneration, their levels increase in the absence of mTORC2.

Methods: Rictor liver-specific knockout (R-LKO) and control mice underwent partial hepatectomy, followed by measurement of TG and FA contents during liver regeneration. FA metabolism was evaluated by analyzing the expression of FA metabolism-related genes and proteins. Intraperitoneal injection of the peroxisome proliferator-activated receptor α (PPAR-α) agonist, p53 inhibitor, and AKT activator was performed to verify the regulatory pathways involved. Lipid mass spectrometry was performed to identify the potential PPAR-α activators.

Results: The expression of FA metabolism-related genes and proteins suggested that FAs are mainly transported into hepatocytes during liver regeneration. The PPAR-α pathway is significantly downregulated in the mTORC2-deficient liver, resulting in the accumulation of TGs. The PPAR-α agonist WY-14643 rescued deficient liver regeneration and survival in mTORC2-deficient mice. Furthermore, lipidomic analysis suggested that mTORC2 deficiency substantially reduced glucosylceramide (GluCer) content. GluCer activated PPAR-α. GluCer treatment in vivo restored the regenerative ability and survival rates in the mTORC2-deficient group.

Conclusions: Our data suggest that FAs are mainly transported into hepatocytes during liver regeneration, and their metabolism is facilitated by mTORC2 through the GluCer-PPAR-α pathway, thereby establishing a novel role for mTORC2 in lipid metabolism.
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http://dx.doi.org/10.1016/j.jcmgh.2022.07.011DOI Listing
August 2022

Precisely controlled polydopamine-mediated antibacterial system: mathematical model of polymerization, prediction of antibacterial capacity, and promotion of wound healing.

Nanotechnology 2022 Aug 2. Epub 2022 Aug 2.

Engineering Research Center of Molecular and Neuro Imaging of the Ministry of education, School of Life Science and Technology, Xidian University, Xifeng Rd. Xilong Sec. No. 266, Xi'an, Shaanxi Province, 710071, CHINA.

In this work, the polydopamine (PDA)-mediated antibacterial system is synthesized to carry out antimicrobial activities in vitro and in vivo. First, to precisely control the surface modification of nanodiamonds (NDs), a mathematical kinetics model of PDA deposition is established, and the conditions of synthesis reaction are discussed including influencing factors such as the concentrations of dopamine, reaction time, and the kinetic constant k1, which is a function of several variables associated with the reaction temperature, light irradiance (especially at ultraviolet wavelengths), pH value and concentration of dissolved O2 in the solution. A simulated visualization demonstrates that the deposition thickness of PDA is positively correlated with temperature and light irradiance, and PDA is easier to deposit in an alkaline solution and will be terminated if the dissolved O2 is insufficient. Then, the precisely controlled thickness of PDA can control the growth of AgNPs, rendering the intensity of Raman peaks increased and providing a predictable antibacterial effect against E. coli in vitro. An optimized antibacterial hydrogel containing NDs-PDA/Ag is prepared and characterized by the Fourier transform infrared spectroscopy and field emission scanning electron microscopy. Finally, the antibacterial experiments to promote wound healing in vivo are performed, which are verified by pathological and immunohistochemical-stained sections. This work provides a theoretical basis of predicting the PDA-assisted surface modification of NDs, giving a divinable antibacterial effect, and promoting wounds healing in vivo.
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http://dx.doi.org/10.1088/1361-6528/ac85f2DOI Listing
August 2022

An acquired phosphatidylinositol 4-phosphate transport initiates T-cell deterioration and leukemogenesis.

Nat Commun 2022 Jul 29;13(1):4390. Epub 2022 Jul 29.

MOE Key Laboratory of Tumor Molecular Biology, Jinan University, Guangzhou, 510632, China.

Lipid remodeling is crucial for malignant cell transformation and tumorigenesis, but the precise molecular processes involved and direct evidences for these in vivo remain elusive. Here, we report that oxysterol-binding protein (OSBP)-related protein 4 L (ORP4L) is expressed in adult T-cell leukemia (ATL) cells but not normal T-cells. In ORP4L knock-in T-cells, ORP4L dimerizes with OSBP to control the shuttling of OSBP between the Golgi apparatus and the plasma membrane (PM) as an exchanger of phosphatidylinositol 4-phosphate [PI(4)P]/cholesterol. The PI(4)P arriving at the PM via this transport machinery replenishes phosphatidylinositol 4,5-bisphosphate [PI(4,5)P] and phosphatidylinositol (3,4,5) trisphosphate [PI(3,4,5)P] biosynthesis, thus contributing to PI3K/AKT hyperactivation and T-cell deterioration in vitro and in vivo. Disruption of ORP4L and OSBP dimerization disables PI(4)P transport and T-cell leukemogenesis. In summary, we identify a non-vesicular lipid transport machinery between Golgi and PM maintaining the oncogenic signaling competence initiating T-cell deterioration and leukemogenesis.
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http://dx.doi.org/10.1038/s41467-022-32104-7DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC9338045PMC
July 2022

miR-4443/MMP2 suppresses the migration and invasion of trophoblasts through the HB-EGF/EGFR pathway in preeclampsia.

Cell Cycle 2022 Jul 28:1-16. Epub 2022 Jul 28.

International Peace Maternity and Child Health Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China.

Preeclampsia (PE) is a pregnancy-associated disease that may cause maternal and fetal morbidity and mortality. The dysregulation of microRNAs (miRNAs) and their potential functions has been an important direction for elucidating the mechanism of preeclampsia in recent years. The present study investigated whether miR-4443 was significantly increased in the placentas of severe preeclamptic patients, and the upregulation of miR-4443 inhibited the migration and invasion of HTR-8/SVneo cells according to transwell assays. Matrix metallopeptidase 2 (MMP2), which is involved in the degradation of extracellular matrix (ECM) components and harbors a miR-4443-binding site within its 3'-UTR as confirmed by a luciferase reporter assay, was identified to be directly inhibited by miR-4443. Moreover, siRNA targeting MMP2 imitated the effects of overexpressed miR-4443 on HTR-8/SVneo cell invasion and migration, whereas rescue experiments showed that MMP2 reversed this inhibitory function of miR-4443. Heparin-binding EGF-like growth factor (HB-EGF), as the downstream gene of MMP2, plays an important role in trophoblast invasion, and we confirmed that the expression of HB-EGF/EGFR pathway-related biomolecules was consistent with MMP2 influenced by upregulating and downregulating miR-4443 and that activated EGFR further transmitted intracellular downstream signaling via the MAPK pathway according to western blot assay. In conclusion, we demonstrated that miR-4443 suppresses the migration and invasion of trophoblasts, and its inhibitory effects are at least partially mediated by the suppression of MMP2. This inhibition might further affect the progression of preeclampsia through the HB-EGF/EGFR pathway, thus providing a new clue on the role of miR-4443 in the pathogenesis of preeclampsia.
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http://dx.doi.org/10.1080/15384101.2022.2103897DOI Listing
July 2022

Tri-Channel Electrochemical Immunobiosensor for Combined Detections of Multiple Exosome Biomarkers of Lung Cancer.

Biosensors (Basel) 2022 Jun 21;12(7). Epub 2022 Jun 21.

State Key Laboratory of High Performance Complex Manufacturing, Central South University, Changsha 410083, China.

Current methods for the early diagnosis of cancer can be invasive and costly. In recent years, exosomes have been recognized as potential biomarkers for cancer diagnostics. The common methods for quantitative detection of exosomes, such as nanoparticle tracking analysis (NTA) and flow cytometry, rely on large-scale instruments and complex operation, with results not specific for cancer. Herein, we present a tri-channel electrochemical immunobiosensor for enzyme-free and label-free detecting carcino-embryonic antigen (CEA), neuron-specific enolase (NSE), and cytokeratin 19 fragments (Cyfra21-1) from exosomes for specific early diagnosis of lung cancer. The electrochemical immunobiosensor showed good selectivity and stability. Under optimum experimental conditions, the linear ranges were from 10 to 10 ng/mL for CEA, 10 to 10 ng/mL for NSE, and 10 to 10 ng/mL for Cyfra21-1, and a detection limit down to 10 ng/mL was achieved. Furthermore, we performed exosome analysis in three kinds of lung cancer. The results showed a distinct expression level of exosomal markers in different types. These works provide insight into a promising alternative for the quantification of exosomal markers in specific diseases in the following clinical bioassays.
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http://dx.doi.org/10.3390/bios12070435DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC9313016PMC
June 2022

Combined cadmium and fluorine inhibit lettuce growth through reducing root elongation, photosynthesis, and nutrient absorption.

Environ Sci Pollut Res Int 2022 Jul 26. Epub 2022 Jul 26.

Key Laboratory of Environment Remediation and Ecological Health, Ministry of Education, College of Environmental and Resource Sciences, Zhejiang University, Zijingang Campus, Yuhangtang Road 866, Hangzhou, 310058, China.

Cadmium (Cd) and fluorine (F) often coexist in environment and are toxic to organisms; however, their combined effects on plants are still not well documented. In this study, the co-effects of Cd and F on germination, biomass, photosynthesis, and nutrients uptake of lettuce were carried out in hydroponic culture. The results showed that the seed germination and seedling biomass decreased with an increase in Cd and F supplementation. The root morphology verified these effects as excess combined Cd and F diminished the root tips and surface area of lettuce, while single Cd and F inhibited the growth by decreasing root length and average diameter, respectively. These effects were also consistence with a reduction in photosynthesis which was mainly regulated by reducing the quantum yield of PS II, electron transport activity, stomatal conductance, intercellular CO concentration, and transpiration rate in response to the pollutants. Moreover, when lettuce exposed to Cd and F stress, the accumulation of several essential elements in shoot decreased. In a sum, the synergistic negative effects of Cd and F on the seed germination and seedling growth of lettuce were observed, and these might be owed to nutrient absorption and translocation in the plant. These findings aid in understanding the harmful effects and specific mechanisms of action of Cd and F on plants.
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http://dx.doi.org/10.1007/s11356-022-22195-6DOI Listing
July 2022

Cadmium exposure causes mitochondrial fission and fusion disorder in the pig hypothalamus via the PI3K/AKT pathway.

Ecotoxicol Environ Saf 2022 Jul 21;242:113880. Epub 2022 Jul 21.

College of Veterinary Medicine, Northeast Agricultural University, Harbin 150000, PR China. Electronic address:

Cadmium (Cd) is the main environmental pollutant causing endocrine and nervous system dysfunction in animals. High doses of Cd cause cytotoxicity, including programmed necrosis and apoptosis, which has aroused widespread concern. Mitochondrial dynamics plays a key role in programmed necrosis and apoptosis of endocrine organs. Nevertheless, there is a lack of information on the relationship between Cd-induced programmed necrosis/apoptosis of the hypothalamus and the mitochondrial fusion-fission balance. Therefore, a hypothalamic injury model of Cd exposure was established by adding 20 mg/kg CdCl to the basic pig diet for 40 days. Analysis of the Cd toxicity mechanism was conducted by inductively coupled plasma mass spectrometry, hematoxylin and eosin staining, the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling assay, and quantitative reverse transcription-polymerase chain reaction, as well as western blot analyses. The results suggested that exposure to Cd inhibited the expression of PI3K and AKT, interfered with the balance of mitochondrial fusion and division, downregulated the expression of Mfn2, Mfn1, and OPA1, and upregulated the expression of Drp1 and Mff, which led to cell apoptosis and programmed necrosis in the pig hypothalamus. This study finds that cadmium exposure leads to mitochondrial fission and fusion dysfunction in porcine hypothalamus via the PI3K/AKT pathway.
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http://dx.doi.org/10.1016/j.ecoenv.2022.113880DOI Listing
July 2022

[Preliminary Study on Drug-Loaded Chondroitin Sulfate-Modified Micelles Targeting Golgi Apparatus in Tumor Cells for the Treatment of Tumor Metastasis].

Sichuan Da Xue Xue Bao Yi Xue Ban 2022 Jul;53(4):619-625

West China Hospital of Stomatology, Sichuan University, Chengdu 610041, China.

Objective: To make preliminary exploration into the Golgi apparatus targeting of chondroitin sulfate-modified micelles (CSmicelles) co-loaded with pirarubicin (THP) and vinorelbine (VRL) in tumor cells, as well as their anti-tumor metastasis effect.

Methods: The cellular uptake efficiency and internalization mechanism of CSmicelles in 4T1 mouse breast cancer cell line were investigated by flow cytometry. Preliminary study of the Golgi apparatus targeting CSmicelles in tumor cells was conducted by co-localization experiment. Then, the effect of CSmicelles co-loaded with THP and VRL (THP+VTL-CSmicelles) on the structure of Golgi apparatus was investigated by GM130 immunofluorescence experiment. Finally, the anti-tumor metastasis ability of THP+VTL-CSmicelles was evaluated by wound healing assay and Transwell migration/invasion assay.

Results: It was found that CSmicelles could significantly increase cellular uptake of drugs. CSmicelles were internalized into cells through clathrin-mediated and caveolin-mediated endocytosis, which was energy-dependent active transport and exhibited substantial ability of targeting Golgi apparatus in tumor cells. THP+VTL-CSmicelles could break down the structure of Golgi apparatus and significantly inhibit the migration and invasion of tumor cells.

Conclusion: THP+VTL-CSmicelles demonstrate high affinity towards Golgi apparatus in tumor cells, exert targeted effects and inhibit tumor cell metastasis, which provides a novel idea and method for the treatment of cancer metastasis.
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http://dx.doi.org/10.12182/20220760103DOI Listing
July 2022

A GC-MS-based untargeted metabolomics approach for comprehensive metabolic profiling of vancomycin-induced toxicity in mice.

Heliyon 2022 Jul 6;8(7):e09869. Epub 2022 Jul 6.

Institute of Clinical Pharmacy and Pharmacology, Jining First People's Hospital, Jining Medical University, Jining 272000, China.

Background: Vancomycin is a glycopeptide antibiotic that is commonly used for severe drug-resistant infections treatment. Application of vancomycin frequently leads to severe ototoxicity, hepatotoxicity, and nephrotoxicity; however, the comprehensive metabolic analysis of vancomycin-induced toxicity is lacking.

Purpose: This study attempted to investigate the metabolic changes after vancomycin administration in mice.

Methods: Experimental mice (n = 9) received continuous intraperitoneal injection of vancomycin (400 mg/kg) every day for 7 days, and mice in control group (n = 9) were treated with the same amount of normal saline. Pathological changes of the kidney were examined using haematoxylin and eosin (HE) staining. A gas chromatography-mass spectrometry (GC-MS) approach was used to identify discriminant metabolites in serum and various organs including the heart, liver, kidney, spleen, cerebral cortex, hippocampus, inner ear, lung, and intestine. The potential metabolites were identified using orthogonal partial least squares discrimination analysis (OPLS-DA). Subsequently, the MetaboAnalyst 5.0 (http://www.metaboanalyst.ca) and Kyoto Encyclopedia of Genes and Genomes database (KEGG, http://www.kegg.jp) were employed to depict the metabolic pathways.

Results: Compared with the control group, the vancomycin induced 13, 17, 27, 22, 16, 10, 17, 11, 10, and 7 differential metabolites in the serum, liver, kidney, heart, cerebral cortex, lung, spleen, intestine, hippocampus, and inner ear, respectively. Further pathway analyses identified that amino acids metabolism, fatty acids biosynthesis, energy metabolism, and lipid metabolism were disrupted after VCM exposure.

Conclusion: Vancomycin affects the metabolism in various organs in mice, which provides new insights for identification of vancomycin-induced toxicity, and facilitate to better understanding of the metabolic pathogenesis of vancomycin.
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http://dx.doi.org/10.1016/j.heliyon.2022.e09869DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC9287194PMC
July 2022

Synchronous Signal Transmission Method of Minimally Invasive Renal Failure Surgery Based on Nano Molecular Image Probe.

J Biomed Nanotechnol 2022 Apr;18(4):1205-1214

Department of Urology, The Central Hospital of Enshi Tujia and Miao Autonomous Prefectrue, Enshi, Hubei, 445000, China.

The synchronous signal transmission can promote the successful completion of minimally invasive renal failure surgery, so the synchronous signal transmission method of minimally invasive renal failure surgery based on nano molecular image probe is studied. The nanoprobe is constructed by loading different signal molecules, photosensitizers, acoustic sensitizers and thermosensitives through the method of double emulsion or film hydration; the near-infrared confocal endoscope molecular image diagnosis and treatment equipment is designed based on the nanoprobe, and the intraoperative highfrequency image is obtained through the diagnosis and treatment equipment; the high-frequency injection energy and signal synchronous transmission method is adopted, and the signal is added to the primary and secondary resonance circuit. In the coupling module, a higher frequency alternating signal is added into the resonant coupling voltage to modulate the signal into the transmission resonance system, which is transmitted from the energy transmission coupling coil to the secondary end, and the injected signal part of the coupling coil is demodulated at the other end to complete the transmission of the signal coupling transmission loop, so as to realize the high-frequency image energy and index data signal of minimally invasive renal failure surgery. The experimental results show that the designed nano molecular image probe has good stability, and the simulated signal transmission waveform is consistent with the transmission waveform of the principle analysis. Applying the proposed method to the minimally invasive surgery of renal failure, it is found that the success rate and image signal transmission efficiency of the minimally invasive surgery of patients 1-5 are higher than 99.5%, and the operation image transmission accuracy is high and the operation effect is excellent.
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http://dx.doi.org/10.1166/jbn.2022.3326DOI Listing
April 2022

Cancer incidence and spectrum among Uygurs in Hotan District in China.

Cancer Epidemiol 2022 Jul 16;80:102217. Epub 2022 Jul 16.

Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center for Cancer, Key Laboratory of Cancer Prevention and Therapy, Tianjin's Clinical Research Center for Cancer, Tianjin 300060, China; Hotan District People's Hospital, Hotan, Xinjiang 848000, China. Electronic address:

Cancer is the leading cause of death in China and a significant public health problem with increasing incidence and fatality rates. The Han nationality is the main ethnic group in China, and many reports on the epidemiology of cancers in Han nationality are published. However no studies report the cancer spectrum of Uygurs which are one of the minority nationalities in China. Hence, we present incidence and mortality numbers of different cancers for the Uygur patients for the period 2018-2020 in Hotan District where Uygur population accounts for 99 %. During the 3-year study period, 2509 new Uygur cancer cases were registered, comprising 774 men and 1735 women. Cervical cancer was the most common, followed by esophageal, breast, gastric and colorectal cancers. The most common cancers in women and men were cervical cancer and esophageal cancer, respectively. In conclusion, the cancer spectrum in Hotan is different from other regions of China and our research revealed the cancer incidence in Hotan, which could help us to take appropriate measures to reduce the incidence rate.
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http://dx.doi.org/10.1016/j.canep.2022.102217DOI Listing
July 2022

Development and characterization of anti-HPV16 monoclonal antibodies for assembly of an HPV16 detection kit.

Biotechnol Appl Biochem 2022 Jul 16. Epub 2022 Jul 16.

Beijing Health Guard Biotechnology Inc., BDA, Daxing District, Beijing, China.

Quality control is very important during the development of 3-valent (16/18/58), 9-valent (6/11/16/18/31/33/45/52/58), and 15-valent human papillomavirus (HPV) vaccines (6/11/16/18/31/33/35/39/45/52/56/58/59/68). All 3-valent, 9-valent, and 15-valent HPV vaccines contain the HPV16 antigen; therefore, a detection method that can specifically identify HPV16 in vaccines is urgently required. This study aimed to develop and characterize monoclonal antibodies to assemble a highly specific HPV16 detection kit. The HPV16 L1 pentameric protein developed as an immunogen was used to prepare monoclonal antibodies. From the pool of prepared monoclonal antibodies, we selected 4G12 and 5A6 to screen and evaluate their subtypes, specificity, neutralizing activity, serum competition, binding affinity, and gene sequencing. After these characterizations, an enzyme-linked immunosorbent assay kit for these monoclonal antibodies was developed, and excellent quality was demonstrated in the assessment of linearity, repeatability, and specificity. The developed detection kit has great potential for wide use in clinical testing and quality control in vaccine production processes.
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http://dx.doi.org/10.1002/bab.2384DOI Listing
July 2022

[Association of sudden sensorineural hearing loss and its prognosis with the brain white matter hyperintensity].

Lin Chung Er Bi Yan Hou Tou Jing Wai Ke Za Zhi 2022 Jul;36(7):523-527

Department of Otolaryngology Head and Neck Surgery,Baoji Municipal Central Hospital.

To explore the relationship between white matter hyperintensity(WMH) and sudden sensorineural hearing loss(SSHL) and evaluate the influence of WMH on the prognosis of SSHL. Fifty hospitalized unilateral SSHL patients and 50 age and gender matched routine physical examination individuals without SSHL history from June 2019 to June 2020 were included for a case-control study. All included subjects underwent 3.0 Tesla cranial magnetic resonance examination, and the Fazekas scale was applied to evaluate periventricular white matter hyperintense(PVWMH) and deep white matter hyperintense(DWMH). Fazekas score and distribution proportions of Fazekas score was compared between SSHL and control. Ordered logistic regression was used to study the relationship between prognosis of SSHL and WMH. The Fazekas score of WMH in SSHL group was significantly higher than that of control group(PVWMH: =0.004, DWMH: =0.010); There was a significant difference in the distribution proportions of Fazekas scores between SSHL and control(PVWMH: =0.036, DWMH: =0.047); The results of ordered logistic regression showed that patients without WMH(Fazekas=0) is an independent predictor of good prognosis in SSHL(=0.025, =12.779). The prevalence of WMH in SSHL patients was higher than that of control. SSHL patients without WMH has a better prognosis than those with WMH.
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http://dx.doi.org/10.13201/j.issn.2096-7993.2022.07.009DOI Listing
July 2022

Carbon tetrachloride exposure induces ovarian damage through oxidative stress and inflammatory mediated ovarian fibrosis.

Ecotoxicol Environ Saf 2022 Jul 8;242:113859. Epub 2022 Jul 8.

Department of Obstetrics and Gynecology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, 430030 Wuhan, Hubei, China; National Clinical Research Center for Obstetrical and Gynecological Diseases, 430030 Wuhan, Hubei, China; Key Laboratory of Cancer Invasion and Metastasis, Ministry of Education, 430030 Wuhan, Hubei, China. Electronic address:

Carbon tetrachloride (CCL) is widely used as a chemical intermediate and as a feedstock in the production of chlorofluorocarbons. CCL is highly toxic in the liver, kidney, testicle, brain and other tissues. However, the effect of CCL on ovarian function has not been reported. In this study, we found that the mice treated with CCL showed decreased ovarian function with disturbed estrus cycle, decreased serum level of 17β-estradiol and the reduced number of healthy follicles. Ovarian damage was accompanied by oxidative stress and the production of proinflammatory cytokines, especially interleukins. The indicators of oxidative stress, 4-Hydroxynonenal (4-HNE), 8-hydroxy-2´-deoxyguanosine (8-OHdG), 3-Nitrotyrosine (3-NT) and malondialdehyde (MDA), and the levels of proinflammatory cytokines IL-1α, IL-1β, IL-6 and IL-11 were increased, while the antioxidants, including superoxide dismutase (SOD), nuclear factor erythroid2-related factor 2 (NRF2) and heme oxygenase-1 (HO-1), were decreased in the CCL group. In the CCL treated group, the results of Sirius Red staining, immunohistochemistry and qPCR indicated that proinflammatory cytokines caused further ovarian fibrosis. And CCL could also promote ovarian thecal cells to secrete inflammatory cytokines, resulting in fibrosis in vitro. In addition, CCL inhibited oocyte development and triggered oocyte apoptosis. In conclusion, CCL exposure causes ovarian damage by strong oxidative stress and the high expression of the proinflammatory cytokine mediated ovarian fibrosis.
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http://dx.doi.org/10.1016/j.ecoenv.2022.113859DOI Listing
July 2022

Effect of 52 % low-sodium salt applied to CM-DASH Diet on atherosclerotic cardiovascular disease risks in patients with hypertension and type-2 diabetes.

Nutr Hosp 2022 Jul 6. Epub 2022 Jul 6.

Health Management Center. Second Affiliated Hospital. Chongqing Medical University.

Introduction: hypertension and diabetes are chronic disorders associated with an increased risk of cardiovascular disease.

Objectives: to evaluate the effect of 52 % low-sodium salt applied to the Chinese-modified DASH (CM-DASH) diet on risk of atherosclerotic cardiovascular disease (ASCVD) in patients with hypertension and type-2 diabetes.

Methods: the low-sodium salt group (LSSG) took 5 g/day of 52 % low-sodium salt plus CM-DASH diet for 8 weeks, while the normal-sodium salt group (NSSG) took the same dose of normal-sodium salt plus CM-DASH diet for 8 weeks. Blood tests, 24-hour urine tests, anthropometric measurements, and 10-year risk of ASCVD prediction were assessed.

Results: compared with baseline, both LSSG and NSSG showed a significant reduction in 10-year risk of ASCVD, but we did not find any statistically significant differences in 10-year risk of ASCVD between LSSG and NSSG.

Conclusions: our study shows that salt limits and DASH diets reduce the risk of cardiovascular disease whereas low-sodium salt containing 52 % sodium chloride did not significantly lower the risk of cardiovascular disease when compared to regular salt. Due to the limitations of the research, additional studies will be necessary to confirm our findings.
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http://dx.doi.org/10.20960/nh.04081DOI Listing
July 2022

Network Pharmacology and Data Mining Approach Reveal the Medication Rule of Traditional Chinese Medicine in the Treatment of Premenstrual Syndrome/Premenstrual Dysphoric Disorder.

Front Pharmacol 2022 21;13:811030. Epub 2022 Jun 21.

College of Traditional Chinese Medicine, Shandong University of Traditional Chinese Medicine, Jinan, China.

Premenstrual syndrome (PMS) is a common disorder that affects women of reproductive age. It is characterized by periodic mental and somatic symptoms such as irritability, depression, and breast pain during the luteal phase. Premenstrual dysphoric disorder (PMDD) is the most severe form of PMS. In recent years, the incidence of PMS/PMDD has been increasing year after year. However, due to the complex symptoms and ambiguous classification of PMS/PMDD, the limitations of present treatments, such as their poor efficacy rate, have become increasingly apparent. With its unique benefits such as syndrome differentiation and high cure rate, traditional Chinese medicine (TCM) has sparked new diagnosing and treating of PMS/PMDD. This study uses data mining methods, and statistical analysis revealed that Xiaoyao San and Chaihu Shugan San were the commonly used TCM to treat PMS/PMDD. A detailed investigation of regularly used single herbs revealed that most TCM is used as cold herbs that penetrate the liver meridian, with predominant bitter, sweet, and pungent flavors. The network pharmacology method analyzes the interactions between diseases, targets, and herbs. Meanwhile, the deep action targets and molecular mechanisms of 10 commonly used herbs for the treatment of PMS/PMDD are studied, revealing that it involves several ingredients, many targets, and different pathways. This interaction provides insight into the mechanism of action of TCM in the synergistic treatment of PMS/PMDD. It is now clear that we can begin treating PMS/PMDD with TCM using the target and mechanism revealed by the abovementioned findings in the future. This serves as an essential reference for future research and clinical application of TCM in the treatment of PMS/PMDD.
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http://dx.doi.org/10.3389/fphar.2022.811030DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC9253672PMC
June 2022

Evaluation of Traumatic Subdural Hematoma Volume by Using Image Segmentation Assessment Based on Deep Learning.

Comput Math Methods Med 2022 28;2022:3830245. Epub 2022 Jun 28.

Department of Clinical Laboratory, HwaMei Hospital, University of Chinese Academy of Sciences, Ningbo 315010, China.

Rapid and accurate evaluations of hematoma volume can guide the treatment of traumatic subdural hematoma. We aim to explore the consistency between the measurement results of traumatic subdural hematoma (TSDH) using a deep learn-based image segmentation algorithm. A retrospective study was conducted on 90 CT images of patients diagnosed with TSDH in our hospital from January 2019 to January 2022. All image data were measured by manual segmentation, convolutional neural networks (CNN) algorithm segmentation, and /2 volume formula. With manual segmentation as the "golden standard," a consistency test was carried out with CNN algorithm segmentation and /2 volume formula, respectively. The percentage error of CNN algorithm segmentation is less than /2 volume formula. There is no significant difference between CNN algorithm segmentation and manual segmentation ( > 0.05). The area under curve of the /2 volume formula, manual segmentation, and CNN algorithm segmentation is 0.811 (95% CI: 0.717~0.905), 0.840 (95% CI: 0.753~0.928), and 0.832 (95% CI: 0.742~0.922), respectively. From our results, the algorithm based on CNN has a good efficiency in segmentation and accurate calculation of TSDH hematoma volume.
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http://dx.doi.org/10.1155/2022/3830245DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC9256325PMC
July 2022

A novel 15.8 kb deletion α-thalassemia confirmed by long-read single-molecule real-time sequencing: Hematological phenotypes and molecular characterization.

Clin Biochem 2022 Jul 2. Epub 2022 Jul 2.

Department of Medical Genetics and Prenatal Diagnosis, Huizhou First Maternal and Child Health Care Hospital, Huizhou, Guangdong, China. Electronic address:

Background: Thalassemia is the most frequent recessive Mendelian inherited monogenic disease worldwide, and is characterized by the impaired synthesis of globin chains due to disease-causing variants in α- or β-globin genes. There are many conventional methods to diagnose thalassemia but all of them have limitations.

Case Report: We present the case of a 37-year-old female with abnormal values of routine hematological indices who was admitted for genetic screening of thalassemia. Genomic DNA was extracted and used for genetic assays covering the known and potential novel genotypes in HBA and HBB genes using a suspension-array system, gap-polymerase chain reaction (Gap-PCR), PCR-reverse dot blot (PCR-RDB) and multiplex ligation-dependent probe amplification (MLPA). Finally, using long-read single-molecule real-time (SMRT) sequencing, we first confirmed the case with a novel 15.8 kb deletion located in the HBA gene (Chr16:163886-179768, GRch38/hg38).

Conclusions: Our results showed that long-read SMRT sequencing has great advantages in the detection of rare α-globin gene variants. This study may provide a reference protocol for the use of long-read SMRT sequencing for the detection of known and potential novel genotypes of thalassemia in the population and improve the accuracy of genetic counseling and prenatal diagnosis.
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http://dx.doi.org/10.1016/j.clinbiochem.2022.06.015DOI Listing
July 2022

Neoadjuvant Therapy for Pancreatic Ductal Adenocarcinoma: Where Do We Go?

Front Oncol 2022 16;12:828223. Epub 2022 Jun 16.

Department of Hepatobiliary Surgery, The First Affiliated Hospital of Dalian Medical University, Dalian, China.

The incidence of pancreatic ductal adenocarcinoma (PDAC) has been on the rise in recent years; however, its clinical diagnosis and treatment remain challenging. Although surgical resection remains the only chance for long-term patient survival, the likelihood of initial resectability is no higher than 20%. Neoadjuvant therapy (NAT) in PDAC aims to transform the proportion of inoperable PDACs into operable cases and reduce the likelihood of recurrence to improve overall survival. Ongoing phase 3 clinical trial aims to validate the role of NAT in PDAC therapy, including prolongation of survival, increased R0 resection, and a higher proportion of negative lymph nodes. Controversies surrounding the role of NAT in PDAC treatment include applicability to different stages of PDAC, chemotherapy regimens, radiation, duration of treatment, and assessment of effect. This review aims to summarize the current progress and controversies of NAT in PDAC.
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http://dx.doi.org/10.3389/fonc.2022.828223DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC9245892PMC
June 2022

Improving the digestive stability and prebiotic effect of carboxymethyl chitosan by grafting with gallic acid: In vitro gastrointestinal digestion and colonic fermentation evaluation.

Int J Biol Macromol 2022 Aug 30;214:685-696. Epub 2022 Jun 30.

College of Food Science and Engineering, Yangzhou University, Yangzhou 225127, Jiangsu, China. Electronic address:

Carboxymethyl chitosan (CMCS) is a useful polysaccharide with potential applications in food, cosmetic and biomedical industries. Nonetheless, CMCS is unfavorable for maintaining intestinal flora balance. In this study, gallic acid (GA) was grafted with CMCS through ascorbic acid/hydrogen peroxide initiated graft copolymerization reaction, producing GA grafted CMCS (GA-g-CMCS). The digestive and fermentative behavior of CMCS and GA-g-CMCS were investigated by using in vitro simulated gastrointestinal digestion and colonic fermentation models. Results showed that the average molecular weight (M) of CMCS gradually decreased during saliva-gastro-intestinal digestion, changing from original sheet-like morphology to porous and rod-like fragments. However, the M and morphology of GA-g-CMCS were almost unchanged under saliva-gastro-intestinal digestion. Meanwhile, the grafted GA moiety was not released from GA-g-CMCS during saliva-gastro-intestinal digestion. As compared with CMCS fermentation, GA-g-CMCS fermentation significantly suppressed the relative abundance of Escherichia-Shigella, Paeniclostridium, Parabacteroides, Lachnoclostridium, Clostridium_sensu_stricto_1, UBA1819 and Butyricimonas, while facilitated the relative abundance of Enterobacter, Enterococcus, Fusobacterium and Lachnospira. In addition, GA-g-CMCS fermentation significantly enhanced the production of short-chain fatty acids. These findings suggested that the digestive stability and prebiotic effect of CMCS were improved by grafting with GA.
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http://dx.doi.org/10.1016/j.ijbiomac.2022.06.170DOI Listing
August 2022

The Synergistic Effect of Ruthenium Complex ∆-Ru1 and Doxorubicin in a Mouse Breast Cancer Model.

Recent Pat Anticancer Drug Discov 2022 Jun 29. Epub 2022 Jun 29.

Guangdong Province Key Laboratory for Biotechnology Drug Candidates, School of Bioscience and Biopharmaceutics, Guangdong Pharmaceutical University, Guangzhou, 510006, P. R. China.

Background: Doxorubicin is a significant drug for the treatment of breast cancer, but its cardiotoxicity is an obvious obstacle. Previously, we confirmed that ruthenium complex (∆-Ru1) and doxorubicin (∆-Ru1/Dox) combination had a synergistic effect in MCF-7 cells, but its biological effect in vivo is unknown.

Purpose: In order to find a way to overcome the toxicity of doxorubicin and build MCF-7 xenograft tumor mouse model to test whether this potential combination has better efficacy and less toxicity.

Methods: The tumor model of nude mice was established to verify the synergistic antitumor effect of the drug combination in vivo. H&E staining was used to detect the toxicity of major organs in mice. Sirius red staining and transmission electron microscopy were used to detect cardiotoxicity. Prussian blue was used to measure iron accumulation in heart tissue. TUNEL staining was used to detect the antitumor effect in vivo. Immunohistochemical staining was used to detect the expression of iron death-related pathway proteins. High-throughput sequencing techniques were used to determine the molecular mechanism of ferroptosis.

Results: Histopathological analysis of tumor tissues indicated that the ∆-Ru1/Dox combination significantly promoted tumor cell apoptosis. Doxorubicin damaged cardiac tissue by inducing fibrosis and iron accumulation, but it was reversed by the ∆-Ru1/Dox combination treatment. Further exploration found that doxorubicin could regulate iron accumulation in the ferroptosis pathway and the expression of lipid peroxidation-related proteins, including upregulation of Tf, DMT1, and HO-1, and downregulation of Nrf2, SLC7A11, and GPX4.

Conclusion: ∆-Ru1/Dox combination synergistically inhibits tumor growth, and it can significantly reduce and alleviate the toxic side effects of doxorubicin, especially the cardiac injury.
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http://dx.doi.org/10.2174/1574892817666220629105543DOI Listing
June 2022

Web-based interventions for pregnant women with gestational diabetes mellitus: a systematic review and meta-analysis protocol.

BMJ Open 2022 06 29;12(6):e061151. Epub 2022 Jun 29.

Women's Hospital, Zhejiang University School of Medicine, Hangzhou, China

Introduction: Gestational diabetes mellitus (GDM) is one of the most prevalent diseases during pregnancy, which is closely associated with many short-term and long-term maternal and neonatal complications and can incur heavy financial burden on both families and society. Web-based interventions have been used to manage GDM because of the advantages of high accessibility and flexibility, but their effectiveness has remained inconclusive. This systematic review and meta-analysis aims to comprehensively investigate the multidimensional effectiveness of web-based interventions for pregnant women with GDM, thereby aiding implementation decisions in clinical settings.

Methods And Analysis: This systematic review protocol strictly adheres to the Preferred Reporting Items for Systematic Review and Meta-Analysis Protocols guidelines. Six electronic databases (PubMed, Web of Science, Cochrane Central Register of Controlled Trials, Embase, CINAHL and PsycINFO) will be comprehensively searched from their inception to 26 January 2022 to identify randomised controlled trials and controlled clinical trials regarding the efficacy of web-based interventions for pregnant women with GDM on glycaemic control, behavioural outcomes, cognitive and attitudinal outcomes, mental health, maternal and neonatal clinical outcomes, and medical service utilisation and costs. Two reviewers will independently conduct the study selection, data extraction and quality assessment. The methodological quality of included studies will be assessed using the Effective Public Health Practice Project assessment tool. The overall meta-analyses for each of the interested outcomes will be performed if the outcome data are sufficient and provides similar effect measures, as well as subgroup analyses for glycaemic control indicators based on the different types of intervention format, interactivity and technology. We will conduct a qualitative synthesis for studies that cannot be quantitatively synthesised.

Ethics And Dissemination: Ethics approval is not required for this review as no human participants will be involved. The results will be disseminated via a peer-reviewed journal or an academic conference.

Prospero Registration Number: CRD42022296625.
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http://dx.doi.org/10.1136/bmjopen-2022-061151DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC9244690PMC
June 2022

ORM 1 as a biomarker of increased vascular invasion and decreased sorafenib sensitivity in hepatocellular carcinoma.

Bosn J Basic Med Sci 2022 Jun 29. Epub 2022 Jun 29.

Department of Pathology, the First affiliated hospital of Dalian Medical University, Dalian, Liaoning Province, China.

This study aimed to clarify the role of Orosomucoid 1 (ORM1) in the development and therapy resistance in hepatocellular carcinoma (HCC). The mRNA expression level of ORM1 was analyzed via integrative analysis of Gene Express Omnibus (GEO) and The Cancer Genome Atlas (TCGA) datasets. The protein expression level of ORM1 in our cohort was determined using immunohistochemistry. Correlation analysis was used to investigate the relationship between ORM1 expression and clinical parameters. The Cell Counting Kit-8 assay was used to clarify the role of ORM1 in HCC malignant behaviors, including cell growth and sorafenib sensitivity, in vitro. The results indicated that ORM1 was significantly downregulated in the hepatic cancer cells compared to that in the non-cancerous cells. However, it was upregulated in microvascular invasion samples, especially in the cancer embolus compared to that in the surrounding tumor cells. Though Kaplan-Meier analysis did not show an association of ORM1 expression with the overall survival rates of HCC patients, univariate analysis indicated that ORM1 expression was highly correlated with tumor grade and stage. An in vitro assay also revealed that downregulation of ORM1 led to the suppression of tumor growth and enhancement of sorafenib sensitivity without epithelial-to-mesenchymal transition (EMT) alteration, which was consistent with our bioinformatic analysis. Hence, ORM1 played a key role in HCC tumorigenesis and may serve as a potential target for the development of therapeutics against HCC in the future.
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http://dx.doi.org/10.17305/bjbms.2022.7268DOI Listing
June 2022

A comprehensive study of the differences in protein expression and chemical constituents in tea leaves (Camellia sinensis var. sinensis) with different maturity using a combined proteomics and metabolomics method.

Food Res Int 2022 Jul 23;157:111397. Epub 2022 May 23.

Key Laboratory of Biology, Genetics and Breeding of Special Economic Animals and Plants, Ministry of Agriculture and Rural Affairs, Tea Research Institute, Chinese Academy of Agricultural Sciences, Hangzhou, Zhejiang 310008, China. Electronic address:

The maturity of tea leaves has a great influence on the flavor quality and commercial price of tea. In this work, a combined proteomics and metabolomics analysis was applied to investigate the differences in protein expression and metabolites among tea leaves with different maturity. Integrated analysis showed that there were significant differences in 112 nonvolatile components related to the pathways of photosynthesis, glycolysis, tricarboxylic acid cycle, and the biosynthesis of amino acids, phenylpropanoids and flavonoids. The bud had higher expression levels of most enzymes related to the biosynthesis of amino acids, phenylpropanoids, and flavonoids, leading to higher levels of amino acids, most flavanols, and procyanidins compared with the leaves. The 1 leaf showed a higher expression level of flavonol synthase, which produces higher levels of flavonol-3-glycosides. This study offers deep insight into the maturity of tea at both the protein and metabolite levels and provides a guideline for tea manufacturing.
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http://dx.doi.org/10.1016/j.foodres.2022.111397DOI Listing
July 2022

Meta-analysis of efficacy and safety of sustained release oxycodone hydrochloride rectal administration for moderate to severe pain.

PLoS One 2022 27;17(6):e0266754. Epub 2022 Jun 27.

Department of Oncology, Fenghua District Hospital of Traditional Chinese Medicine, Ningbo, Zhejiang Province, China.

Objective: This study aims to evaluate the efficacy and safety of oxycodone hydrochloride (OxyContin) rectal administration in cancer pain patients. This is geared towards providing the research evidence for a novel route of OxyContin administration.

Methods: Relevant randomized controlled trials (RCTs) were searched in electronic databases, including PubMed, Cochrane Library, Web of Science, EMBASE, China National Knowledge Infrastructure (CNKI), Chinese Scientific Journal Database (VIP database), Wanfang Data Knowledge Service Platform, and Chinese Biomedical Literature Database (CBM). Moreover, unpublished academic data were obtained by contacting the colleague, professor, or Institute of Traditional Chinese Medicine. The RCTs of transrectal Oxycodone administration of sustained-release tablets for moderate and severe pain patients were searched in the databases from inception to December 2020.

Results: According to the inclusion criteria, a total of 8 RCTs were included, with a total of 648 patients. Meta analysis results showed that there was no statistically significant difference in the efficacy of moderate to severe pain control between the rectal administration group and the oral administration group (RR = 1.04, 95%CI: 0.99-1.10, p = 0.13>0.05). At the same time, the incidence of adverse reactions in the rectal administration group was low. In terms of constipation, the rectal administration group was less than the oral administration group, with a statistically significant difference (RR = 0.43, 95%CI: 0.31-0.58, p< 0.00001). In terms of nausea and vomiting, the rectal administration group was less than the oral administration group, and the difference was statistically significant(RR = 0.30, 95%CI: 0.21-0.42, p<0.00001). In terms of sleepiness, there was no significant difference between the two groups(RR = 0.54, 95%CI: 0.26-1.15, p = 0.11>0.05). In terms of dizziness, there was no statistically significant difference between the two groups (RR = 0.43, 95%CI:0.27-0.68, p = 0.31>0.05). In terms of dyuria, there was no statistically significant difference between the two groups (RR = 0.37, 95%CI: 0.02-7.02, p = 0.51>0.05). In terms of KPS scores there was no significant difference was noted between the rectal and oral administration groups (RR = 1.04, 95%CI: 0.89-1.21, p = 0.63>0.05).

Conclusion: In summary, we found no significant differences in efficacy between rectal administration of OxyContin and oral administration. Thus, rectal administration should be considered in managing cancer pain among patients with difficulty in oral OxyContin administration.

Prospero Registration Number: CRD42021209660.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0266754PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC9236234PMC
June 2022

Isolation of Leydig cells from adult rat testes by magnetic-activated cell sorting protocol based on prolactin receptor expression.

Andrology 2022 Sep 11;10(6):1197-1207. Epub 2022 Jul 11.

Zhejiang Provincial Key Laboratory of Anesthesiology, Department of Anesthesiology, The Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, Zhejiang, China.

Background: The primary function of testicular Leydig cells (LCs) is to produce testosterone (T). In vitro culture of the cells represents a very important approach to study androgen production and its regulations. Various methods have been developed for the enrichment of the cells from the testes. However, getting cells in large numbers with high purity and viability is still challenging. Here, we describe a new way to isolate LCs from rat testes in large quantity with high purity and viability.

Methods: Enzymatic digested testicular cells from adult rats were labelled with prolactin receptor (PRLR) antibody. The positive cells were isolated by magnetic-activated cell sorting (MACS) protocol. Purified LCs were tested in vitro for their steroidogenic (T production) and non-steroidogenic (25-OH-vitamin D production and Insl3 and Cyp2r1expressions) functions in the presence of Luteinizing Hormone (LH) for up to 24 h.

Results: Reanalysis of scRNA-seq data indicates that Prlr expression is highly specific in LCs of adult rat testis. MACS procedure based on PRLR expression was able to isolate LCs with very high yield (about 10 cells/testis), high purity (about 95%) and viability (> 93%). Purified LCs retained high steroidogenic and non-steroidogenic functions in responding to maximal LH stimulations, with more than 10-fold increases in T production in 3 h and 42% and 103% increases in Insl3 and Cyp2r1 expressions in 24 h.

Discussion And Conclusion: We have established an excellent way to purify high quality LCs from adult rat testis that can serve as a useful tool to study the physiology, pharmacology and toxicology of the cells in vitro.
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http://dx.doi.org/10.1111/andr.13211DOI Listing
September 2022

Clinical features and next-generation sequencing landscape of essential thrombocythemia, prefibrotic primary myelofibrosis, and overt fibrotic primary myelofibrosis: a Chinese monocentric retrospective study.

J Cancer Res Clin Oncol 2022 Jun 22. Epub 2022 Jun 22.

Department of Hematology, The Fourth Affiliated Hospital of Zhejiang University School of Medicine, N1 Shangcheng Road, Yiwu, Zhejiang, People's Republic of China.

Objective: Since prefibrotic primary myelofibrosis (pre-PMF) was recognized as a separate entity in the 2016 revised classification of MPN differed from essential thrombocythemia (ET) or overt fibrotic primary myelofibrosis (overt PMF), it has been a subject of debate among experts due to its indefinite diagnosis.

Methods: We retrospectively reviewed the clinical parameters, haematologic information, and genetic mutations of patients who were diagnosed with myeloproliferative neoplasms (MPNs) according to the WHO 2016 criteria in China, including 56 ET patients, 19 pre-PMF patients, and 43 overt PMF patients.

Results: Pre-PMF patients exhibited higher leukocyte counts [14.2(6.0-28.1) × 10/L vs 9.6(4.0-55.0) × 10/L, P = 0.003], LDH values [307(233-479)U/L vs 241(129-1182)U/L, P < 0.001], onset ages [67(32-76) years vs 50(16-79) years, P = 0.006], a higher frequency of splenomegaly(47.4% vs 16.7%, P = 0.018) and hypertension (57.9 vs 23.2%, P = 0.005) than ET patients. On the other hand, pre-PMF patients had higher platelet counts [960(500-2245) × 10/L vs 633(102-1720) × 10/L, P = 0.017], haemoglobin levels [152(115-174)g/L vs 119(71-200)g/L, P = 0.003], lower LDH values [307(233-479)U/L vs 439(134-8100)U/L, P = 0.007] and a lower frequency of splenomegaly(47.4 vs 75.6%, P = 0.031) than overt PMF patients. Next-generation sequencing landscape was performed in 50 patients, revealed the frequency of EP300 mutations was significantly increased in pre-PMF patients compared with ET and overt PMF patients (60 vs 10 vs 15.79%, P = 0.033), and WT1 was more often overexpressed (WT1/ABL1 copies ≥ 1.0%) in patients with overt PMF than in those with ET or pre-PMF(54.55 vs 16.67 vs 17.65%, P = 0.009). In terms of outcome, male sex, along with symptoms including MPN10, anaemia (haemoglobin < 120 g/L), thrombocytopenia (platelet count < 100 × 10/L), leucocytosis (leukocyte counts > 13 × 10/L), high LDH value (> 350U/L), splenomegaly, WT1 overexpression(WT1/ABL1 copies ≥ 1.0%), KMT2A, ASXL1 and TP53 mutations, indicated a poor prognosis for PMF patients.

Conclusion: The results of this study indicated that a comprehensive evaluation of BM features, clinical phenotypes, haematologic parameters, and molecular profiles is needed for the accurate diagnosis and treatment of ET, pre-PMF, and overt PMF patients.
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http://dx.doi.org/10.1007/s00432-022-04067-1DOI Listing
June 2022
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