Disease bio-banks are an important strategic resource for medicine development. Such a bank with complete information and high information sharing enhances the competitiveness of medicine and promote the development of translational medicine. The present development of such banks is found with such setbacks as weak government leadership and standardization, as well as lack of a sharing mechanism. Therefore medical institutions should strengthen their standardization and informationization of disease bio-banks, which deserve sufficient government policy support and guidance. These efforts will help preserve our rich clinical resources, in addition to their integration and sharing for medicine advancement.

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 establish chronic stress model of depression in adolescent rats and to examine the effects of different antidepressant treatment on depression and anxiety-related behaviors.Methods Male Wistar rats were given 21-day chronic mild stress (CMS) during their adolescence (postnatal day 30～50, PND30～50).During stress period, rats were treated with fluoxetine (10 mg/kg i.p.) or imipramine (10 mg/kg i.p.), respectively.After stress, rats were tested for behavioral observation using body weight gain, saccharine test, open field and elevated plus-maze (EPM).Results Compared with control/vehicle (n=10) group, stress/vehicle (n=11) group displayed lower weight gain, saccharine preference index and the number of rearing in open field (P<0.05).Antidepressant fluoxetine, but not imipramine reversed anhedonia and the decrease of the number of rearing induced by stress.In addition, compared with early adolescent(PND29) rats, late adolescent (PND52) rats in control/vehicle group exhibited less open arm entries and open arm time, more closed arm time in EPM (P<0.05).Rats in stress/vehicle group showed more open arm entries and less closed arm time than controls(P<0.05).Both fluoxetine and imipramine had no effects on such changes.Conclusions Stress can induce the depression-like behavior in adolescent rats.Fluoxetine, but not imipramine,can effectively reverse anhedonia induced by stress.However, Both antidepressants have no significant effects on stress-induced decrease in developmental increment of anxious behavior during adolescence.These data suggest that chronic mild stress have complicated effects on depressive and anxious behavior in adolescent rats.

Objectiv e:To investigate the effect of different culture conditions on the differentiation of Treg and Th17 to lay a foundation for exploring the methods to reverse the immune tolerance induced by tumor microenvironment.Methods:The IL-6 gene was cloned and stablely transferred into the tumor cell line expressing TGF-β.The conditioned mediums ( CM) were prepared by collecting the culture supernatants of tumor cell lines with or without IL-6 expression and used in the in vitro culture of peripheral blood mononuclear cells ( PBMC ) .The changes of Treg and Th17 in PBMC treated with different CM were detected with flow cytometry ( FCM) .Results:The expression of TGF-βin BEL-7402 was higher than that in HepG2.Thus the BEL-7402 was selected for preparation of cell line stablely transfected with IL -6 gene.ELISA detection confirmed the effective expression of IL -6 by the identified cell lines.It was showed that the Treg increased in PBMC treated with culture supernatants of tumor cells .However,the presence of IL-6 reversed the increase of Treg and promoted the differentiation of Th 17.Conclusion: The culture supernatants of tumor cells increases the proportion of Treg.However,the presence of IL-6 in this CM can reverse the increase of Treg and raise the proportion of Th 17.

BACKGROUND:With the wide application of bone repair scaffold in the field of medicine, nano-hydroxyapatite (nHA) and silk fibroin (SF) both of which have good biological properties have become research hotspots in recent years.OBJECTIVE:To study the feasibility of nHA/SF composite materials loaded with recombinant human bone morphogenetic protein 2 (rhBMP-2) to restore the initial stability of the spinal segment in rabbits.METHODS: Thirty-six male health New Zealand rabbits were randomly divided into three groups, followed by preparation of spinal instability models. Autogenous iliac bone, nHA/SF composite, and rhBMP-2/nHA/SF composite were implanted into the L4/5 spinal segment in autologous bone group, nHA/SF group and rhBMP-2/nHA/SF group, respectively. X-ray exmaination was performed at 12 weeks postoperatively, and then the animals were killed for gross observation. The stability of the fusion segments was tested through a tensile machine. Histologically, bone graft fusion at the surgical site was observed.RESULTS AND CONCLUSION:(1) Findings from the gross specimen observation showed that the specimens at the fusion site presented with a hard texture. Obvious signs of fusion were visible in the autologous bone group, followed by the rhBMP-2/nHA/SF group, while no signs of fusion were detected in the nHA/SF group. (2) At 12 weeks postoperatively, a large number of trabecular bones grew into the boundary between the vertebral body and the iliac crest graft block in the autologous bone group. A little trabecular bone was found in the boundary in the nHA/SF group, while a lot of trabecular bone tissues were found in the boundary in the rhBMP-2/nHA/SF group. In accordance with the standard of fusion, there were 10, 3, and 9 rats in the autologous bone, nHA/SF and rhBMP-2/nHA/SF groups, respectively. (3) The range of motion of the spine in the rhBMP-2/nHA/SF showed no statistical difference from that in the autologous bone group, but was significantly higher than that in the nHA/SF group (P < 0.05). (4) Osseous connection was found around the bone graft in the autologous bone and nHA/SF groups, but no mature bone tissue was visible in the latter group. In the rhBMP-2/nHA/SF group, a large number of new capillaries was found and permeated into the spinal tissues. In summary, nHA/SF composite materials loaded with rhBMP-2 possess good biocompatibility, mechanical properties and bone induction ability, which can rebuild the initial stability of the spine in a short time.

Objective To analyze the impacts of two different loading methods on the sterilization of glass feeding bottle in hospitals,to confirm the best loading sterilization methods and provide the acceptable product for the clinical department.Methods By purposive sampling the glass feeding bottles sterilized between December 2013 and December 2014 in Central Sterile Supply Department,the First People's Hospital of Shangqiu,were chosen.We divided them into experimental group and control group with each 5 groups (120 bottles).The control group applied medical storage tank to load and sterilize.The experimental group sterilized through innovative stainless steel hollow with cover.Both groups were deposited in the storage rack in the clean area with air 300 000 stage air purification and temperature and humidity meeting the national standards.The professional worker supervised this two items at the same time in validity.In the period of validity,professional personnel implemented visual inspection,a magnifying glass detection and bacterial culture at the same time.Results The visualization and magnifying glass detection qualified rate of 5 packages (120 bottles) in the control group was 75.0％,sterilization quality rate 95.0％ by biological cultivation compared with 95.8％ and 100.0％ in the experimental group (P ＜ 0.05).Conclusions The usage of wrapping tool with innovative hollow stainless steel can ensure the effectiveness and the integrity of the glass feeding bottles,can reduce the medical cost and the occurrence of hospital infection.

Objective To study the killing effect of γδT cells on different tumor cells by inducing and expanding γδT cells in vitro.Methods Collected the peripheral venous blood of 20 healthy volunteers recruited at Xinqiao Hospital of Third Military Medical University from January to March 2018.Peripheral blood mononuclear cells (PBMCs) were induced and expanded using zoledronic acid combined with interleukin-2 (IL-2) to obtain γδT cells.The cell purity was detected on day 0,7,10,14 and 16 using flow cytometry.The killing activity was detected by CCK-8 kit.Cells cultured on the 14th day were used as effector cells,and breast cancer cell BCap-37 and liver cancer cell Bel-7402 were used as target cells,and the cell killing activity was detected at the effective target ratio (E ∶ T) of 5 ∶ 1,10 ∶ 1 and 20 ∶ 1 respectively.Results The purity of γδT cells were respectively (3.35 ± 1.32) ％,(50.76 ± 5.76) ％,(80.43 ± 4.53) ％,(90.56 ± 3.34) ％,(89.54 ± 4.42) ％ on day 0,7,10,14,16,with significant difference (F =18.431,P =0.012).The efficiency of γδT cells killing BCap-37 tumor cells were (31.3 ± 2.0) ％ (E ∶ T =5 ∶ 1),(48.7 ± 1.6) ％(E ∶T=10∶1),(71.3 ±2.4)％ (E ∶T=20 ∶ 1) respectively,with significant difference (F=16.724,P =0.016),further comparison between each two groups:10 ∶ 1 vs.5 ∶ 1 (P =0.013),20 ∶ 1 vs.5 ∶ 1 (P =0.017),20∶1 vs.10 ∶1 (P =0.011).The killing rate increased with the increase of target ratio.The efficiency of γδT cells killing Bel-7402 tumor cells were (34.5 ± 2.0) ％ (E ∶ T =5 ∶ 1),(52.4 ± 1.9) ％(E ∶ T =10 ∶ 1),(74.5 ± 1.6) ％ (E ∶ T =20 ∶ 1) respectively,with significant difference (F =18.253,P=0.013),further comparison between each two groups:10 ∶ 1 vs.5 ∶ 1 (P=0.015),20 ∶ 1 vs.5 ∶ 1 (P=0.012),20 ∶ 1 vs.10 ∶ 1 (P =0.015).The killing rate increased with the increase of target ratio.Conclusion We can obtain high purity γδT cells using zoledronic acid combined with IL-2 induced PBMCs,and the amplified γδT cells have significant killing effect on BCap-37 and Bel-7402 tumor cells.

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 explore the effects of WeChat platform on treatment compliance,self-efficacy and quality of life in patients with atrial fibrillation.Methods A total of 84 patients with atrial fibrillation who were treated in Yancheng City No.1 People's Hospital from January 2015 to August 2016 were selected as the subjects. They were randomly divided into observation group and control group according to random number table, each group of 42 cases. The patients in the control group received routine health education, while patients in the observation group received health education through the WeChat platform. The treatment compliance, self-efficacy and quality of life were compared between the two groups.Results After the intervention, scores of the observation group in taking medication as order, IRN prescribed medication monitoring, regular exercise, treatment and prevention of bleeding, thrombosis complications and hazards, giving up smoking and drinking and diet control were significantly higher than that of the control group, and the differences were statistically significant (t=3.323, 3.247, 3.975, 2.646, 3.524, 4.669, 4.178;P<0.05). Before and after nursing intervention,the improving degree of patients in the observation group were higher than that in the control group in 10 dimensions of movement scale, disease information, getting help, physician communication, general disease management, doing housework, social and recreational activities, symptom management, shortness of breath management, and depression control, and the differences were statistically significant (t=4.885, 5.641, 4.863, 4.245,3.887, 3.768, 3.795, 3.314, 3.323, 3.856; P<0.05). The scores of patients in the observation group in physiological function, psychological function, social function, environmental adaptation ability and total quality of life scores improved significantly than that in the control group (t=5.965, 4.863, 3.746, 4.247, 4.769;P<0.05).Conclusions The application of WeChat platform in the health education for patients with atrial fibrillation can significantly improve the treatment compliance,self-efficacy and quality of life.

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.