Objective:To study the expression of stomatin-like protein 2(STOML2)in oral squamous cell carcinoma(OSCC)tissue and the effects of STOML2 on the tumorigenicity of OSCC cells(OSCCCs)in vitro and in vivo,and the related mechanism.Methods:The protein expression of STOML2 in OSCC and adjacent tissues of 56 patients was detected.OSCCCs SCC-15 were divided into 2 groups.Stom12-siRNA plasmid was transfected into the cells of experimental group and Mock-siRNA plasmid was transfected into the cells of control group.The mRNA and protein expression of STOML2,CDK4 and P16 in the cells was detected by qPCR and Western blot respectively.The cell cycle of the cells was detected by flow cytometry,and the proliferation of the cells was detected by CCK8 asay.The tumorigenicity of the cells was detected by subcutaneous tumor model in nude mice.Results:The positive rate of STOML2 in OSCC and adjacent tissues was 92.86％(52/56)and 8.93％(5/56)respectively(P＜0.001).After siRNA transfection,STOML2 mRNA expression in SCC-15 cells of experimental group and control group was(0.43±0.09)and(1.23±0.19),STOML2 protein ex-pression was(0.52±0.11)and(0.94±0.17)respectively.CDK4 expression was(0.33±0.13)and(1.18±0.17),P16 expression was(0.93±0.12)and(0.29±0.03),respectively.In CCK8 assay the absorbance of SCC-15 cells in experimental group and control group was(1.11±0.24)and(2.19±0.28),in flow cytometry the percentage of cells in G2/M phase was 35.72％±5.33％and 18.65％±3.71％(P＜0.05),respectively.In vivo test showed that the volume(μm3)of subcutaneous transplanted tumor was 1 192.07 ±250.9 and 2 280.5±600.1,the weight(g)of mice was 0.65±0.30 and 1.62±0.40,respectively.Conclusion:STOML2 expression increases in OSCC,STOML2 affects the tumorigenic ability of OSCCCs in vitro and in vivo by regulating P16 related pathways.

OBJECTIVE To evaluate the levels of evidence and analyze the rationality for off-label use of tacrolimus in the kidney diseases in adults. METHODS By systematically reviewing the drug instructions, clinical guidelines and medical literature of tacrolimus in the kidney diseases in adults, screening the highest level of evidence, and an evaluation table based on evidence-based medical for off-label use of tacrolimus in the kidney diseases in adults was established. Based on the evaluation table, collect adult patients from the Department of Nephrology in Zhongda Hospital Southeast University who had a medication history of off-label use of tacrolimus from January 1, 2021 to December 31, 2022, and evaluated the rationality of application for tacrolimus. RESULTS A total of 19 indications for off-label use of tacrolimus in the kidney diseases in adults were listed, and their recommended levels were determined based on evidence. Among them, the recommended levels for common types of kidney diseases were higher. In addition, 194 adult patients with off-label use of tacrolimus were selected from the Department of Nephrology in Zhongda Hospital Southeast University, and their application recommended levels with "strongly recommended""moderately strongly recommended""weakly recommended" and "not recommended", were 67.0%, 12.9%, 15.0% and 5.1%, respectively. CONCLUSION Tacrolimus has a wide range of indications for off-label use in the kidney diseases in adults, but the evidence for most common types of kidney diseases are relatively sufficient and their use are reasonable.

To explore the mechanism of tea albino variation and high theanine formation, 'Fuyun 6' and a new theanine-rich tea cultivar 'Fuhuang 2' were as materials in this study, pigment content, metabolome and transcriptome of the two cultivars were analyzed by ultramicroelectron microscopy, widely targeted metabolomics, targeted metabolomics and transcriptomics. The results showed that five catechins, theobromine, caffeine, and 20 free amino acids, including theanine, glutamine, arginine, etc., were identified by targeted metabolomics. The amino acid content of 'Fuhuang 2' was significantly higher than that of 'Fuyun 6', and the theanine content was as high as 57.37 mg/g in 'Fuhuang 2'. The ultrastructure of leaves showed that the chloroplast cell structure of 'Fuhuang 2' was fuzzy, most of the grana lamellae were arranged in disorder, with large gaps, and the thylakoids were filiform. The determination of pigments showed that compared with 'Fuyun 6', the contents of chlorophyll A and B, carotenoids, flavonoids and other pigments of 'Fuhuang 2' decreased significantly, some important pigment-related-genes, such as chlorophyllase (CLH), 9-cis-epoxycarotenoid dioxygenase (NCED), flavonoid 3β-hydroxylase (F3H) and flavonoid 3', 5'-hydroxylase (F3'5'H) were significantly changed. Compared with 'Fuyun 6', 'Fuhuang 2' identified 138 significantly changed metabolites (SCMs) and 658 differentially expressed genes (DEGs). KEGG enrichment analysis showed that SCMs and DEGs were significantly enriched in amino acid biosynthesis, glutathione metabolism and TCA cycle. In general, the albino phenotype of 'Fuhuang 2' may be caused by a deficiency in photosynthetic proteins, chlorophyll metabolism genes and chlorophyll content. The accumulation of high theanine in 'Fuhuang 2' may be due to the low nitrogen consumption in yellowed leaves and the lack of carbon skeleton, amino and nitrogen resources are stored more effectively, resulting in the up regulation of metabolites and related gene expression in the amino acid synthesis pathway, theanine has become a significant accumulation of nitrogen-containing compounds in yellowed leaves.
Camellia sinensis/genetics* ; Chlorophyll A/metabolism* ; Plant Proteins/genetics* ; Plant Leaves/chemistry* ; Chlorophyll/metabolism* ; Transcriptome ; Flavonoids/metabolism* ; Amino Acids/genetics* ; Tea ; Mixed Function Oxygenases/metabolism* ; Nitrogen/metabolism*

Objective:To observe the expression of S100A8 in plasma exosomes, microvesicles (MV), plasma and vitreous in patients with diabetic retinopathy (DR), and verify it in a diabetic rat model, and to preliminarily explore its role in the occurrence and development of DR.Methods:A case-control study. From September 2018 to December 2019, a total of 73 patients with type 2 diabetes, hospitalized patients undergoing vitrectomy, and healthy physical examinations in the Tianjin Medical University Eye Hospital were included in the study. Among them, plasma were collected from 32 patients and vitreous fluid were collected from 41 patients, which were divided into plasma sample research cohort and vitreous sample research cohort. The subjects were divided into simple diabetes group (DM group), non-proliferative DR group (NPDR group) and proliferative DR group (PDR group) without fundus changes; healthy subjects were regarded as normal control group (NC group). In the study cohort of vitreous samples, the control group was the vitreous humor of patients with epimacular membrane or macular hole. Plasma exosomes and microvesicles (MVs) were separated using ultracentrifugation. Transmission electron microscopy, nanometer particle size analyzer and Western blot (WB) were used to characterize exosomes and MVs. The mass concentration of S100A8 was determined by enzymelinked immunosorbent assay. Eighteen healthy male Brown Norway rats were divided into normal control group and diabetic group with 9 rats in each group by random number table method. The rats of diabetes group were induced by streptozotocin to establish diabetic model. Five months after modeling, immunohistochemical staining and WB were used to detect the expression of S100A8 in the retina of rats in the normal control group and the diabetes group. t test was used for the comparison of measurement data between the two groups. Single-factor analysis of variance were used for the comparison of multiple groups of measurement data.parison of measurement data between the two groups. Single-factor analysis of variance were used for the comparison of multiple groups of measurement data. Results:Exosomes and MVs with their own characteristics were successfully separated from plasma. The concentrations of plasma exosomes and vitreous S100A8 in the PDR group were higher than those in the NPDR group, DM group, NC group, and the difference was statistically significant ( P=0.039, 0.020, 0.002, 0.002, P<0.000,<0.000). In the plasma sample cohort study, It was not statistically significant that the overall comparison of the S100A8 mass concentrations of plasma and plasma MV in the four groups of subjects ( F=0.283, 0.015; P=0.836, 0.996). Immunohistochemical staining showed that retinal ganglion cells, bipolar cells, cone rod cells and vascular endothelial cells in the diabetic group all expressed S100A8 protein. Compared with the normal control group, the expression level of S100A8 in the retina of the diabetic group increased, and the difference was statistically significant ( t=8.028, P=0.001). Conclusions:The level of S100A8 protein in circulating exosomes increases significantly with the severity of DR in patients with type 2 diabetes. S100A8 may be an influential factor in the inflammatory environment of DR and a potential anti-inflammatory therapeutic target.

Objective:To investigate the protein expression changes of human umbilical cord mesenchymal stem cells (hUCMSCs) modified with pigment epithelium-derived factor ( PEDF) gene mediated by lentivirus. Methods:The hUCMSCs were divided into the control group and experimental group.Cells in the control group were normal hUCMSCs and the cells in experimental group were hUCMSCs with PEDF modification.The proteins from the two groups were collected and processed by FASP method.Samples were fractionated by liquid chromatography and analyzed by tandem mass spectrometry, and SWATH mode was applied.Differential protein analysis, Gene Ontology (GO) analysis and Reactome pathway enrichment analysis were performed. Results:A total of 5 361 quantified proteins were detected in this experiment, of which 432 proteins were differentially expressed (fold change>1.5, P<0.05). There were 219 of the 432 proteins up-regulated, including serpin family F member 1 (SERPINF1) (PEDF), DEAD-box helicase 59 (DDX59) and thrombospondin 1 (THBS1), etc., whereas 213 proteins were down-regulated, including collagen type Ⅰ alpha 1 (COL1A1), COL18A1, etc.GO analysis indicated that the differential proteins were mainly involved in fibrinolysis, extracellular structure organization, regulation of transporter activity, biosynthetic process of secondary alcohol and cholesterol, coenzyme metabolic process and regulation of peptidase activity, etc.Reactome pathway analysis showed that the differential proteins were mostly involved in regulation of insulin like growth factor (IGF) transport and uptake by IGF binding protein, post-translational protein phosphorylation, extracellular matrix organization, metabolism of steroids. Conclusions:After gene modification with PEDF mediated by lentivirus, the expression of many proteins in hUCMSCs were changed. PEDF gene modification can alter the structure of extracellular matrix and regulate the expression of proteins associated with cell proliferation, self-renewal and multipotency.

Objective    To compare the efficacy of percutaneous closure guided by transthoracic echocardiography or angiography in the treatment of patent ductus arteriosus (PDA). Methods    Literature databases such as CNKI, VIP, Wanfang Database, PubMed, Cochrane Library were searched for collecting published literatures on percutaneous closure for PDA guided by transthoracic echocardiography and angiography, retrieval time limit was up to April 2019. Two evaluators independently screened the literature, extracted the data and evaluated the quality according to inclusion and exclusion criteria. The collected data were analyzed by RevMan 5.3 software. Results    Eight studies were included finally, with a total sample size of 681 cases. Meta-analysis showed that there was no statistical difference in the operative success rate between the echocardiography group and the angiography group (RR=0.99, 95%CI 0.97- 1.01, P=0.40). Postoperative complications were less in the echocardiography group than those in the angiography group (RR=0.26, 95%CI 0.11-0.59, P=0.001).The operation time (P<0.000 01), amount of intraoperative radiation (P< 0.000 01), exposure time (P<0.000 01), hospitalization days (P<0.000 01) and hospitalization costs (P<0.000 01) in the echocardiography group were less or shorter than those in the angiography group, and the difference was statistically different. Conclusion    Compared with angiography-guided, transthoracic echocardiography-guided percutaneous closure for PDA is a safe and effective method with less trauma, lower cost, and can replace angiography as one of the guiding methods for PDA.

Objective: To determine the changes of protein expressions in human retinal microvascular pericytes (HRMPCs) stimulated with high glucose by using quantitative proteomics, which provides new clues for future investigation of diabetic retinopathy (DR). Methods: HRMPCs were divided into two groups.The cells in control group were cultured in DMEM basic medium with 25 mmol/L glucose, while the cells in high glucose group were cultured in DMEM medium with 35 mmol/L glucose.The amount of living cells was measured by cell counting kit-8(CCK-8). The proteins were collected from the two groups and then were digested with trypsin.Peptides of 2 μg were injected into the time of flight-mass spectrometer and the acquisition mode was DDA.The results were further analyzed using bioinformatics software. Results: CCK-8 results showed that the absorbance (A₄₅₀) of HRMPCs in high glucose group was 0.75±0.04, which was significantly lower than 0.91±0.05 in control group (t=5.784, P=0.000 2). In total, 1 972 proteins were identified and 54 of them were significantly different between the two groups (fold change >1.5). Among them, 13 proteins were up-regulated, including CTNNB1 and CTBP2; while 41 proteins were down-regulated, including SQSTM1 and HMGCS1.The differentially expressed proteins were mainly involved in citric acid cycle and aerobic respiration. Conclusions The expressions of many proteins in HRMPCs change under the stimulation of high glucose, which may influence the respiration and the ATP production of cells and eventually induce the loss of pericyte.

Objective To detect the protein expression change in the proliferation of human retinal microvascular endothelial cells (hRMECs) stimulated with 4-Hydroxynonenal (4-HNE).Methods hRMECs were in a logarithmic growth phase, and then were separated into 4-HNE-stimulated group and negative control group. The concentration of 4-HNE included 5, 10, 20 and 50 μmol/L in 4-HNE-stimulated group, while the negative control group was added in the same volume of ethanol (the solvent of 4-HNE). Then the cells were stimulated with 4-HNE for 24 hours following by CCK-8 kits incubating for 4 hours to detect absorbance. It was found that 10 μmol/L 4-HNE had the most obvious effect on the proliferation of hRMECs. Therefore, the cellular proteins from 10 μmol/L 4-HNE-stimulated group and negative control group were acquired and prepared by FASP sample preparation method. Data independent acquisition was used for data acquisition, and the GO analysis and pathway enrichment were performed for analysis of differentially expressed proteins. Results CCK-8 kits detection results showed that theA value of the 10 and 20 μmol/L 4-HNE-stimulated groups were significantly higher than negative control group and 5 μmol/L 4-HNE-stimulated group (F=25.42, P<0.01), while there were no differences between 10 and 20 μmol/L 4-HNE-stimulated groups, and theA value of 50 μmol/L 4-HNE-stimulated groups was lower than negative control. A total of 2710 quantifiable proteins were identified by peoteomics,and 118 proteins were differentially expressed (fold change>1.5,P<0.05). Seventy-two proteins were up-regulated after 4-HNE stimulation, whereas 46 proteins were down-regulated. Particularly, the expressions of Heme oxygenase-1, Sulfoxdoxin-1, Heat shock protein A1B, Thioredoxin reductase-1, Glutathione reductase, ATPase and prothrombin were increased when cells were added in 4-HNE, whereas the expressions of apolipoprotein A1 and programmed cell death protein 4 were decreased. These differentially expressed proteins were mainly involved in the biological processes such as oxidative stress, cell detoxification, and ATPase-coupled membrane transport.Conclusion After stimulated with 4-HNE, the oxidative stress, cell detoxification, and ATPase-coupled membrane transport protein expression may change in hRMECs in order to regulate oxidative stress and growth situation.

Objective: To observe the expression of retinal proteins in experimental autoimmune uveitis (EAU) mice and to explore the possible molecular mechanism of autoimmune uveitis. Methods: Twelve female C57BL/6J mice were randomly divided into model group and normal control group, 6 mice in each group.In the model group, the EAU model was established by subcutaneous injection of human interphotoreceptor retinoid-binding protein (IRBP) 651-670.The fundal change of EAV mice was assessed by direct ophthalmoscope, OCT and histopathological staining.At 18 days after immunization, the retinas of the two groups were taken for retinal protein extraction, protein restriction enzyme digestion, mass spectrometry detection, data analysis, and bioinformatics analysis.This study was approved by the experimental animal Ethics Committee of Tianjin Medical University Eye Hospital (TJYY2018070113). The feeding and use of experimental animals follow the ARVO statement. Results: The EAU mouse model was successfully established.At 10 days after immunitation, the retina of EAV mouse was damaged.At 18 days after immunization, retinal edema and infiltration of inflammatory cells into vitreous were observed.Proteomic results showed that a total of 4 458 proteins were identified in this study, of which 522 were differentially-expressed proteins (fold change>1.5, P<0.05). Among these differentially-expressed proteins, 147 proteins including Stat1 and Stat3 were up-regulated and 375 proteins including Gucy2f and Rho were down-regulated.These differentially-expressed proteins were closely related to platelet activation, integrin signaling, T cell activation, the phototransduction cascade, Toll-like receptor and so on. Conclusions EAU is related to the abnormal expression of Stat1, Stat3 and other proteins, as well as the abnormal regulation of platelet activation and integrin signal pathway.

Objective? To establish a WeChat online system supporting continued nursing care for patients receiving enteral bladder augmentation and to evaluate its effects on continued nursing care for patients with neurogenic bladder in spinal cord injury who received enteral bladder augmentation. Methods? Totally 60 patients with low-compliant neurogenic bladder in spinal cord injury who received enteral bladder augmentation and were discharged from Zhejiang Provincial People's Hospital between March 2016 and March 2018 were selected by convenient sampling, and divided into the observation group (n=30) and the control group (n=30) according to the random number table. Patients in the control group received conventional nursing care after discharge, while patients in the observation group received nursing care after discharge via the WeChat online support system. The follow-up duration was 3 months. Bladder self-management compliance, urinary catheter-associated complications, bladder function, quality of life and negative emotions at discharge and 3 months after discharge were compared between the two groups. Results? The score of bladder self-management compliance, anxiety score, depression score, and scores of physical function, role function, emotional function and social function in quality of life in the observation group were better than those in the control group 3 months after discharge (P＜0.05); the rates of urinary tract infection and urethral injury of the observation group were 3.3% and 6.7%, while those of the control group were 26.7% and 33.3%, respectively; the incidence rate of urinary catheter-associated complications of the observation group was lower than that of the control group (P＜0.05); and the parameters of bladder function such as bladder volume, bladder compliance, residual urine volume and maximum detrusor pressure of the observation group were better than those of the control group (P＜ 0.05). Conclusions? The WeChat online support system can improve patients' bladder self-management compliance, reduce the urinary catheter-associated complications, enable the recovery of bladder function, ameliorate their negative emotions, and improve their quality of life.