Background Rapamycin(RAPA)is a specific inhibitor of the mammalian target of rapamycin (mTOR).Researches showed that RAPA inhibits the proliferation of lens epithelium cells(LECs)and tumor cells and induces apoptosis of tumor cells.To investigate whether rapamycin has the inhibitory effect on retinal pigment epithelium(RPE)cells is very important for the prevention and management of proliferative vitreoretinopathy (PVR).Objective This study was to investigate the effects of RAPA on the proliferation and apoptosis of human RPE cells in vitro.Methods Human RPE cells(D407 strain)were cultured and passaged and then divided into regular culture group(blank control group),DMSO control group(0.1‰ DMSO +regular culture),and different concentrations RAPA-treatment groups(5,10,20,40,80,160,320 nmol/L).The proliferation(A490)of human RPE cells was detected using MTT,and the inhibitory rates of RAPA on the proliferation of RPE cells were calculated and compared among different groups at 12,24 and 48 hours.The apoptosis rates of the cells were analyzed among various groups by Hoechst staining after 12,24,48 hours.Results The inhibitory rates of RAPA on RPE cells were significantly different among various groups(F=484.451,P＜0.01)and evidently elevated in 20-320 nmol/L RAPA groups compared with DMSO control group(P ＜ 0.01).The inhibition of RAPA on the cells was considerably enhanced as the lapse of time(F=232.262,P＜0.01)with more dominant effects in 24 and 48 hours compared to 12 hours after addition of RAPA(P＜0.05-0.01).Compared with blank control group and DMSO control group,the apoptotic rates of the cells were evidently increased in 12,24,48 hours in 10 nmol/L RAPA group(all P＜0.05),and higher cellular apoptotic rates were found in 20-320 nmol/L RAPA groups(all P＜0.01).The alteration of cellular apoptotic rate showed a gradually incremental trend as the acting time of RAPA(F =625.584,P＜0.01).Karyorrhexis and mass-like density staining and chromatin substance were seen in RPE cells under the fluorescence microscope in ≥ 10 nmoL/L RAPA groups.Conclusions RAPA suppresses the proliferation and induces the apoptosis of human RPE cells in concentration-and time-dependent manner in vitro.

Objective To investigate the effects of the overexpression of HO-1 induced by CoPP in the donor on the survival of transplanted allogeneic islets of rats and the mechanism.Methods (1) Brown Norway rats were randomly divided into control group, and CoPP-induced group receiving intraperitoneal injection of CoPP (2.5 mg/kg) at 3rd and 1st day prior to islet isolation.By using the cytoimmunofluorescenee and Western blot, the expression of HO-1 in isolated islets was detected.The insulin level in the supernatant of the cultured islets stimulated with glucose was determined by ELISA.(2) Lewis male rat diabetic models were established by a single intravenous injection of alloxan, and then randomly divided into CoPP group and control group.Islets were transplanted under the left kidney capsule of each diabetic recipient.The survival time after transplantation, and pathological changes following rejection of the islet grafts were analyzed.Results The HO-1 was highly expressed in the islets isolated from CoPP-treated rats by cytoimmunofluorescence and Western blot.After stimulation with 16.7 mmol/L glucose, the insulin concentration in Copp-treated and Copp-untreated groups was (46.60± 1.13) and (19.01 ± 1.49) mIU/L respectively (P<0.05).The insulin concentration in Copp-treated and Copp-untreated groups in islets stimulated with 5.6 mmol/L glucose was (15.65 ± 0.89) and (12.28 ± 0.89) mU/L respectively (P>0.05).The stimulated index in Copp-treated and Copp-untreated groups was (2.98 ± 0.10) and 1.55 + 0.01 respectively (P< 0.05).The survival time of islets allograft in Copp-treated and Copp-untreated groups was separately (12.20±5.67) and (5.60± 1.14) days respectively (P<0.05).Histological analysis revealed the presence of more islands of insulin-positive cells and considerably fewer lymphocytes or inflammatory infiltration than the controls.Conclusions CoPP could induce the HO-1 expression of islets, and improve their function.Over-expression of HO-1 in islets could prolong survival time of islets allograft.

Background In recent years,with the contiunous progress of the refractive surgery,the operation skill of phakic intraocular lens(PIOL)implantation for correcting extreme high myopia,astigmatism,farsightedness have made greater progression,and its security,effectiveness in clinical attract much more attention. Objective This study was to evaluate the efficacy,safety and stability of Toric Implantable Collamer Lens(TICL)for extreme high myopic astigmatism. Methods This retrospective case series included 33 eyes of 27 patients from May 2008 to February 2009.A TICL was intraocularly implanted via a 3 mm clear corneal incision after paraocular anesthesia.Patients were examined preoperatively and followed-up at 1 day,1 week,1 month,3,6,12 and 18 months postoperatively.The examinations included uncorrected visual acuity,best corrected visual acuity(BCVA),slit lamp examination,refraction,intraocular pressure,endothelial cell morphometry,etc.The written informed consent was obtained from each patient before any medical procedure. Results The uncorrected visual acuity in 96.97% eyes was equal or improved after operation in comparison with BCVA of preoperation.The spherical refraction was within-1.00 D-+0.25 D.The cylinder refraction was within-1.00 D-0 D.The axial deviation of TICL within 10 degree was 93.94%(31/33).No significant differences were found in the intraocular pressure and endothelial cell morphometry between preoperation and postoperation(intraocular pressure:F=3.35,P=5.49;endothelial cell morphometry:t=1.835,P=0.082).The visual acuity and refraction were stable during the follow-up.Astigmatic axial rotation required surgical intervention on one eye.One eye occurred high intraocular pressure because of bigger TICL diameter.The intraocular pressure returned to normal after TICL was exchanged.No cataract occurred during the follow-up duration. Conclusion TICL implantation appears to be an effective,safe and reliable method for extreme high myopic astigmatism.

Objective To validate the performance of six enzymatic glycated albumin reagents and evaluate their clinical application.Methods The performance of six enzymatic glycated albumin reagents(labled as A,B,C,D,E,F) from Beijing Jiuqiang Co, Beijing Lideman Co,Ningbomeikang Co, Beijing Haomai Co, Sichuan Maike Co and Asahi Kasei Co were assessed on Olympus AU5800 automatic biochemistry analyzer.According to the standard of CLSI,the precision,interference and linear correlation of these reagents were assessed.To assess the accuracy of GA% ,we used GA standard material whose value had been assigned using ID-LC/MS method provided by ReCCS.To do the method comparison and determine the consistency of assay, 50 fresh serum samples of T2DM outpatient and 80 fresh serum samples of apparently healthy people in Jan 2016 were tested using six kits.According to the EP28-A3C protocol, the reference range for GA%was validated in 122 apparently healthy individuals undertaking medical examination from January to February 2016 in PUMC.Results The precision,and the ability of anti-interference of the six reagents were good.The accuracy percentage deviation of six reagents was-19.3%-9.2%.The correlation coefficient of domestic reagents A to E and imported reagents F in the determination of GA% was 0.966-0.999, the average absolute bias was 7.0%-10.4%.The coincidence rate of A-E and F in determining abnormal GA% was between 88.5% and 96.9%.The coincidence rate was increased after switching to the reference range for preliminary clinical evaluation.Conclusion Six GA enzymatic kits used in automatic biochemical analyzer have high precision and strong anti-interference ability.Accuracy still needs to be improved.

OBJECTIVETo analyze the characteristics of CDR3 of TCRbeta on CD8+ T cells in chronic hepatitis B patients.

METHODSEight patients with chronic hepatitis B (ALT more than 2 ULN) were enrolled in this study. CD8+ T cells were isolated from peripheral blood. RT-PCR was proformed to amplify the CDR3 of TCRbeta, and the PCR products were sequenced and analyzed.

RESULTSThe chronic hepatitis B patients showed obvious clonal expansion of T cell, and three perturbation patterns of T cell expansion were showed in the CDR3 of TCRbeta, including monoclonicity, oligoclonicity and skewed peak patterns. The number of perturbation families of CD8+ subpopulation was significantly higher than that of CD8- subpopulation (10.6+/-4.7 vs. 4.1+/-3.1, t = 6.619, P less than 0.01). In 3 out of 8 patients, the number of perturbation families of CD8+ subpopulation was also higher than that of PBMCs without depleting CD8+ subpopulation.

CONCLUSIONSThe characteristics of CDR3 of TCRbeta may help to understand the inflammatory response in CHB patients.

Adult ; CD8-Positive T-Lymphocytes ; immunology ; Complementarity Determining Regions ; genetics ; Genes, T-Cell Receptor beta ; Hepatitis B, Chronic ; blood ; immunology ; Humans ; Male ; Receptors, Antigen, T-Cell ; immunology ; Young Adult

OBJECTIVETo investigate the short-term spontaneous fluctuation of viral load in patients with chronic hepatitis B (CHB) and explore the related factors in treatment naive CHB patients during immune clearance phase.

METHODSA total of 123 treatment naive HBeAg-positive CHB patients with ALT>2 × ULN were enrolled in this study. Paired serum samples were obtained at the first and second visits with an interval of less than 4 weeks. The levels of quantitative HBV DNA (Roche COBAS), quantitative HBsAg, ALT and AST were analyzed. Liver biopsy specimen were collected within 4 weeks and evaluated using Knodell and Ishak histological scoring system.

RESULTSOf the 123 patients, 93 (75.6%) and 30 (24.4%) had HBV DNA fluctuation ≤ 0.5 Log IU/ml and >0.5 Log IU/ml, respectively. Binary logistic multivariate regression analysis identified Knodell necroinflammation score and HBV DNA level as the factors related to HBV DNA fluctuation. Patients with Knodell necorinflammation score ≥ 10 or HBV DNA<7 Log IU/ml had significantly higher rates of HBV DNA fluctuation>0.5 Log IU/ml (50.0% vs 18.3%, P=0.042; 42.9% vs 20.6%, P=0.030).

CONCLUSIONTreatment naive CHB patients in immune clearance phase show short-term spontaneous fluctuation of HBV DNA, and nearly 25% of the patients have HBV DNA fluctuation >0.5 Log IU/ml. Such fluctuation is related to liver inflammation and quantity of HBV DNA.

Adult ; DNA, Viral ; blood ; Female ; Hepatitis B virus ; growth & development ; Hepatitis B, Chronic ; virology ; Humans ; Liver ; physiopathology ; Male ; Viral Load ; statistics & numerical data ; Young Adult

Objective To investigate the changes of prevalence of hyperuricemia ( HUA) and its correlations with blood glucose and lipid in healthy adults receiving physical examination at Peking Union Medical College Hospital (PUMCH) from 2012 to 2017. Meth-ods An observational approach was adopted for the data analysis.The test results of uric acid (UA),fasting blood glucose (FBG),to-tal cholesterol (TC),triacylglycerol (TG),low density lipoprotein cholesterol (LDL-C),high density lipoprotein cholesterol (HDL-C), creatinine (Cr) and Urea of 399 089 cases (206 881 males and 192 208 females) at PUMCH from January 2012 to December 2017 were collected and statistically analyzed.Results The total prevalence of HUA was 17.4% in which the prevalence of males was signif-icantly higher than that of females (25.6% vs 8.5%,χ2＝20 234.850,P＜0.01).During the years of 2012 to 2017,the prevalence of HUA was 26.5%,24.7%,28.6%,23.9%,24.8% and 24.5% in males,and 13.8%,6.3%,7.9%,6.1%,6.2% and 6.8% in females for each year respectively.The prevalence of HUA in males aged 18 to 64 years old was significantly higher than that in the age-matched fe-males (all P＜0.05).However, the prevalence of HUA in males aged≥65 years old was similar to the age-matched females.There was no statistically significant difference of HUA prevalence between males and females aged ≥65 in 2013,2015,2016 and 2017 ( χ2＝1.792,0.017,1.440 and 0.205 respectively;all P＞0.05).The percentages of hyperlipidemia in both males and females of HUA group were higher than those of non-HUA group respectively (all P＜0.01).The percentage of hyperglycemia in males of non-HUA group was higher than that of HUA group,but the percentage of hyperglycemia in females of non-HUA group was lower than that of HUA group ( all P＜0.01).High levels of TC,TG and FBG were risk factors of HUA with increased OR values in increased concentrations of TC,TG and FBG,respectively.Conclusion During the recent 6 years, in healthy adults receiving physiced examination at PVMCH, the preva-lence of male HUA diagnosed was at overall high level,but the prevalence of female HUA was in decreasing and relatively stable trend. Hyperlipidemia and hyperglycemia should be the risk factors of HUA.

Osteofascial compartment syndrome (OFCS) is clinically common and is well known to orthopedic surgeons.Clinicians attach great importance to OFCS because of its severe clinical consequences,and decompression of fascial compartment is often performed in emergency treatment.This article reviews the literature on the threshold of fascial compartment decompression proposed by many scholars in the past and discusses the problems in the clinical diagnosis of acute compartment syndrome,especially the inconsistent pressure thresholds as the indication for emergency decompression surgery.By observing calf fractures patients with tension blister,we found that the pressure of fascia decreased sharply upon the appearance of blisters.Meanwhile,the swelling gradually subsided as well as the clinical manifestations of pain and parasthsia.In view of the uncertainty of various thresholds of fascial decompression and self-decompression,the concepts of myofascial self-release law and muscle-swelling syndrome were first proposed.The author believes that when intracompartmental pressure rises to a point,some unknown mechanisms of fascia can achieve self-decompression.Therefore,no compartment syndrome will take place.We also emphasize that the ' muscle-swelling syndrome'should be strictly distinguished from the soft tissue necrosis caused by crush syndrome and acute limb vascular injury,so as to provide more precise treatment.We believe that without external restrictions such as casts,splints and compression bandages,the muscle-swelling syndrome can achieve self decompression by releasing the pressure in the compartment through tension blisters,and there is no need for fasciotomy.

Objective To study the relationship among rs505802 in SLC22A12,rs6855911,rs737267,rs12498742, rs7442295, rs734553, rs16890979 in SLC2A9 genetic polymorphisms and hypouricemia in Ningxia.Methods 6 056 subjects were collected by multistage,stratified random cluster sampling method in October and November in 2011 in Ningxia Hui autonomous region, 98 subjects with hypouricemia were selected.According to gender and age,84 controls were selected.Physical examination and laboratory biochemical index test were conducted for the study population.T test was used to compare general clinical data and biochemical indexs between two groups. SNPs were detected by Sequenom Mass ARRAY technology.By x2test,we compared the frequencies of the geno-type and allele in each group.Samples representativeness was confirmed through the Hardy-Weinberg inspection. Results The levels of TC, LDLC, and Cr in the patients were lower than those in the control group(P<0.05). There were significant differences in the distribution of A,G allele frequencies of SLC2A9 gene rs7442295 between two groups.The risk of hypouricemia in patients with A/A genotype was lower than that of A/G genotype(Pc<0.05),indicating that A>G mutation was associated with hypouricemia.Conclusions Polymorphisms of SLC2A9 gene rs7442295 are significantly correlated with hyporuricemia in Ningxia.