Background Some aboard studies showed that polyatomic pathway play an important role in the microsvasculopathy of type 2 diabetes,and sorbitol dehydrogenase (SDH) gene is involved in the pathogenesis of diabetic retinopathy(DR).There is no relevant research at home up to now.Objective This study was to investigate the correlation of sorbitol dehydrogenase (SDH) G-888C gene polymorphism with diabetic retinopathy (DR) in type 2 diabetes mellitus.Methods The SDH genotypes were determined by polymerase chain reactionreaction fragment length polymorphism (PCR-RFLP) in 187 patients with diabetes and 123 normal contrels.Patients were divided into two groups depending on the presence or absence of DR(DR:n=118;NDR:n=69).The frequencies of SDH genotype and allele were assayed and compared among these groups.This study was approved by Ethic Committee of Guangdong General Hospital.Written informed consent was obtained from each individual prior to any relevant medical procedure.Results The disease course in the DR group was significantly longer than that in NDR group(t=2.070,P=0.042),and other clinical features in both groups were non-significant (all P>0.05).The genotype frequencies in the DR group,NDR group and normal control group were 24.6%,8.7% and 8.1%,respectively,and frequencies of the G allele were 42.4%,25.4% and 29.7%,showing statistically significant differences among these three groups.The GG genotype and G allele frequencies were significantly higher in the DR group than those in the NDR group and normal control group (GG:P=0.007,P=0.001;G:P=0.001,P=0.004).There were no significant differences in the frequencies of the GG genotype and G allele between the NDB group and normal control group ( P>0. 05) as well as the proliferative DR group and non-proliferative DR group (P>0.05).Conclusion SDH G-888C gene polymorphism is associated with the development of diabetic retinopathy in southern Chinese.

?AIM:To evaluate the objective visual quality of patients who underwent corneal cross-linking for the keratoconus using double-pass analysis system.? METHODS: Advanced keratoconus patients who underwent UV - riboflavin corneal cross - linking from January to July 2015 were included. The outcomes of their objective scattering index ( OSI ) , predicted visual acuity ( VA ) , the cut - off frequency of modulation transfer function ( MTF cut- off ) , the Strehl ratio ( SR ) were compared before and 6mo after corneal cross-linking.?RESULTS: A total of 13 patients ( 16 eyes ) were included. There was no statistically significant difference between pre- and 6mo postoperative data in uncorrected visual acuity, best corrected visual acuity, refractions and mean value of Sim-k (P>0. 05). Non-invasive average tear film break up time ( NIAvg-BUT ) detected by the Sirius system decreased after corneal cross-linking ( P<0. 05 ) . Using double - pass analysis system, no statistically significant change was found in MTF cut off, Strehl Ratio, OSI before and after treatment(P>0. 05). Tear Film Analysis Mean OSI increased at 6mo postoperatively (P<0. 05).? CONCLUSION: The subjective visual quality isn’t effected by corneal cross-linking. The tear stabilities of patients are influenced by these operations at 6mo postoperatively. More observations on long-term effect are needed to be taken in the future.

Objectives To investigate the factors influencing long-term quality of life (QOL)of patients with first-episode ischemic stroke(IS)and partial side limb dysfunction.Methods 126 cases of the first ischemic stroke patients with partial side limb dysfunction admitted to hospital from November 2012 to July 2013 in Shanghai First People's Hospital were selected.General information of patients was collected and evaluated using NIH Stroke Scale(NIHSS)and Modified Rankin Scale(MRS).Following-up was conducted using NIHSS,MRS,stroke specific quality of life (SS-QOL) and Hamilton Depression Scale (HAMD) during 2-3 years after stroke Factors related to long-term QOL were screened by single factor analysis,then factors influencing QOL by multiple stepwise regression analysis.Results The average QOL score of patients with first IS partial side limb dysfunction was(181.5 ± 46.4)points scale.In each single scoring,the scale was(10.4±4.3)points in energy,(10.8±4.2)points in family roles,(21.8±6.2)points in language,(22.5 ±7.6) points in activity,(18.5±6.3)points in mood,(10.1±4.4)in personality,(19.2±5.9)points in self-care ability,(16.1 ± 7.0) points in social roles,(11.65 ± 3.31) points in thinking,(19.1 ± 6.6) points in upper limb function,(14.0 ± 2.0) points in visual acuity,(8.7 ± 4.1) points in work.The multiple regression analysis showed that the influencing factors of life quality included long-term NIHSS evaluation(β=-0.376,P＜0.01),HAMD evaluation(β=-0.375,P＜0.01),hypertension history(β=-0.178,P＜0.05).HAMD evaluation affected 9 fields of QOL,and NIHSS as long-term evaluation affected 6 fields of QOL.Conclusions Predictive factors for declined QOL of first-episode ischemic stroke patients with partial side limb dysfunction include higher NIHSS score,severer depression after long-term stroke and combined hypertension in the early stage.These indicate that recovering of body structure and function injury,control of hypertension,effective prevention and treatment of post-stroke depression maybe important measures for improving life quality of ischemic stroke patients with partial side limb dysfunction.

Background The pathogenesis and development of cataract is associated with oxidative stress-induced apoptosis of human lens epithelial cells(LECs).BH3-only protein is a factor that can initiate apoptosis,and thus the apoptotic process is probably related to the activation of the c-Jun N-terminal kinase(JNK).However,the relationship between oxidative stress-induced apoptosis of human LECs and the JNK pathway remains to be illuminated.Objective This study was to investigate the effects of the JNK/c-Jun pathway and its target gene,Bim (Bcl-2 interacting mediator of cell death)and PU M A(p53 up-regulated modulator of apoptosis),on oxidative stressinduced apoptosis of human LECs.Methods The human LECs cell line(HLEC-B3)was cultured and passaged in DMEM with 10％ fetal bovine serum in vitro.Confluent cells were incubated in 24 well plates and divided into 4 groups.Hydrogen peroxide(H2O2)(50 μmol/L)was used to treat the cells for 4,8 or 12 hours,and cells without H2O2 treatment served as the control group.Apoptosis was detected using Hoechst 33258 staining and quantified by counting the number of cells with pyknotic nuclei.In addition,confluent cells were seeded in 6 well plates,and Western blot and RT-PCR were used to detect the expression of the caspase-3,c-Jun,Bim and PUMA proteins and their mRNA in HLEC-B3,respectively.The JNK/c-Jun pathway inhibitors,CEP11004 or SP600125,were added into cultured media with H2O2,and cells treated with DMSO or H2O2 only served as negative and positive control groups.The expression of the p-JNK,JNK,p-c-Jun,c-Jun,Bim,PUMA proteins was detected by Western blot and apoptosis was assayed using Hoechst 33258 staining.200 pmoL/L of Bim or PUMA small interference RNA(siBim or siPUMA)fragments were transfected into the cells for 24 hours,respectively,and H2O2 was then used to treat the cells for 8 hours.The expression of the Bim and PUMA protein and their mRNA in the cells was detected by Western blot and RT-PCR,respectively.Results After H2O2 treatment in HLEC-B3 cells for 4,8,or 12 hours,the rates of apoptosis were 4.30％±1.15％,27.08％±0.74％ and 46.59％±0.91％,showing a significant difference among them (F=1909.433,P=0.000),and those of the 4,8,12 hour groups were significantly increased in comparison to the control group(P =0.049,0.000,0.000).Compared to untreated cells,the levels of expression of the JNK,Bim,PUMA proteins and their mRNA in HLEC-B3 cells were significantly elevated.After the addition of CEP11004 or SP600125,the expression of these protein and mRNA in HLEC-B3 cells in the presence of H2O2 was significantly weaker than that in the DMSO control group(P =0.000,0.000).After the tranfection of siBim or siPUMA,the apoptosis rates of the H2O2 treated groups were significantly higher than those in the Bim-/-or PIMA-/-group (P＜0.05).Conclusions H2O2 can activate the JNK/c-Jun pathway and up-regulate the expression of its target genes Bim and PUMA in human LECs in a time-dependent manner.Inhibiting the JNK/c-Jun pathway and interfering with the expression of Bim and PUMA can protect human LECs against oxidative stress-induced apoptosis.

Objective To investigate the relationship between V-raf murine sarcoma viral oncogene homolog B1 (BRAF)v600E mutation and radioactive iodine (RAI) uptake in distant metastases from papillary thyroid cancer(PTC).Methods From January 2011 to December 2012,40 PTC patients (21 males,19 females,average age 39.8 years) with distant metastases were recruited and divided into mutation group and wild group according to the BRAFv600E mutation in primary lesions.The clinical,pathological and serological differences were compared between the two groups.The relationship between BRAFv600E mutation and RAI uptake capability in distant metastases from PTC,as well as its relationship with Tg change after 131I treatment were investigated.Statistical analysis was performed with two-sample t test,x2 test or Fisher exact test.Results The BRAFv600E mutation rate was 30.0％ (12/40) in patients with metastases from PTC.There was no significant difference in clinical,pathological and serological features between mutation group (n =12) and wild group (n=28; t:from-0.533 to 1.728,x2:from-1.951 to 1.088,all P＞0.05).Twelve PTC patients had no RAI uptake in the distant metastases,of which 10 belonged to mutation group (83.3％,10/12) and 2 belonged to wild group (7.1％,2/28; x2=19.734,P＜0.05).BRAFv600E mutation group was more likely to have no RAI uptake in the distant metastases.Tg change after 131I treatment in 30 patients were analyzed.In the wild group,Tg level decreased in 66.7％ (14/21) patients,stabilized in 19.0％ (4/21)and increased in 14.3％ (3/21)patients.While there was no decrease of Tg in the mutation group (0/9).Two patients had increased Tg level and 7 patients (with no RAI uptake) kept stable in mutation group.Conclusions Due to poor RAI uptake capability in PTC patients with BRAFv600E mutation,both primary and metastatic sites may have poor response to 131I treatment.Molecular detection of BRAFv600E mutation might be helpful for choosing PTC with distant metastases and predicting the effect of 131 I treatment.

Objective To investigate the prevalence and risk factors of hyperuricemia (HUA) in rural residents of Gaoyou City,Jiangsu Province.Methods A total of 4504 rural residents were investigated in a cross-sectional study.Questionarre survey and physical examination were used for the stndy.Fatsing venous blood samples were collected for biochemical examination.T-test,chi-square test for multiple-group data comparison and logistic regression analysis were used for statistical analysis.Results The prevalence of HUA was 11.9％,15.7％ in male,which was significantly higher than 8.6％ in the female (P＜0.01),the risks of HUA in male was 1.98 times higher than in female.The average value of serum uric acid in this population was higher than.that of female.The prevalence of HUA in female increased with age.Non-conditional logistic regression analysis showed WHR,Cr,Tg,hypertension were the independent risk factors of male patients with HUA.Age,BMI,Cr,Tg,hypertension were independent risk factors of HUA in female.Conclusion The prevalence of HUA can be affected by age and sex.Weight control,keeping blood pressure and blood lipid profiles in normals levels are important for the prevention of HUA.

Objective To observe homing of mesenchymal stem cells (MSCs) in immune organs and inflammatory joints in collagen induced arthritis(CIA) rats. MethodsRats MSCs were isolated and expanded from bone marrow cells by density gradient centrifugation and adhering to the culture cell walls, and the phenotypes were assessed by flow cytometry. MSCs were labeled by PKH-26 and Brdu. Sixty-four rats were randomly divided into normal group and CIA group. Every 8 rats were sacrificed at 3, 11, 30, 42 days after transplantation of MSCs. At the end of the experiment, the specimens of thymus gland, spleen, ankle joints were exposed, fixed, decalcified, wrapped and cut into slices. Confocal laser scanning microscope and immunohistochemical method were used to observe migration and distribution of MSCs in different organs. Independent samples group t test with SPSS 12.0 software package was used for statistical analysis. ResultsIt was found that allogenic MSCs could stay in spleen, thymus gland and joints of CIA rats for a relatively long period (42days). Forty-two days after transplantation of MSCs, the average grey scale values of spleen and thymus gland in CIA group(37.5±8.8, 29.9±5.9 respectively) were significantly higher than the normal group(16.0±2.3,13.2±4.3 respectively), the average grey scale values of ankle joints in CIA group 78±8 was significantly lower than the normal group 93±14(P<0.05). ConclusionIt has been found that MSCs can stay in the injured tissue and organs preferentially.

Objective To investigate the phenotype and function of CD8+T cells cultured with SK-OV-3. Methods Transwell coculture experiments were conducted in 24-well plates with inner wells to separate CD8+ T cells and SK-OV-3. After 5 days of culture, CD8+ T cells were washed, and 1 × 106 cells were collected for Foxp3, CD25, CD28, CTLA-4 and GITR mRNA analysis and 2 × 106 cells were collected to detect expression of Foxp3, CD25, CD28, CTLA-4 and GITR in CD8+ T cells by flow cytometry. CD8+T cells that cultured alone or with SK-OV-3 were added at ratios of 1∶0 , 1∶1 , 1∶5 , 1∶10 , and 0∶1 to na?ve CD4+ T cells in 96-well plates. All wells were cultured with the presence of irradiated PBMCs and anti-CD3 antibody. After 72 h, [3H]-thymidine was added for 16 h prior to the determination of proliferation by scintillation counting. Results Compared with CD8+ T cells cultured without SK-OV-3 , the expression of Foxp3 and CTLA-4 was increased and CD28 expression was decreased in CD8+ T cells cultured with SK-OV-3 (both P < 0.001). We found that CD8+T cells cultured with SK-OV-3 significantly suppressed the na?ve CD4+ T cell proliferation induced by the anti-CD3 stimulation in a dose-dependent manner. In contrast, CD8+ T cells cultured without SK-OV-3 did not suppress na?ve CD4+ T cell proliferation. Conclusion Ovarian cancer cell can induce the suppressive CD8+Treg, which is an important link of the immunosuppressive microenvironment in ovarian cancer.

OBJECTIVETo establish a simple but effective method of laser scanning confocal microscopic imaging for Ca2+ oscillations of pancreatic acinar cells in adult mice.

METHODSPancreatic acinar cells from adult Kunming mice were isolated acutely with collagenase, and then loaded with fluo-4-AM, a Ca2+ indicator. A laser scanning confocal microscope armed with 488 nm laser was employed to record the dynamic fluorescent signals in-time and synchronously while acetylcholine (ACh) was added in the pancreatic acinar cells.

RESULTS(1) The classic pancreatic acinar cell Ca2+ oscillations were induced by a certain concentration of ACh (100 nmol/L) successfully and steadily, which could be blocked by atropine completely. (2) Plasmic Ca2+ oscillations from different parts of one acinar cell were usually with different amplitudes and almost the same frequencies. But both of amplitudes and frequencies were different among different cells. (3) The acinar cell Ca2+ oscillations were induced by ACh in a concentration-dependent manner.

CONCLUSIONThe laser scanning confocal microscopic imaging for adult mouse pancreatic acinar cell Ca2+ oscillations was established successfully. The features of being easy to use, direct to see lively, high efficiency and good flexibility make it a popular tool for researchers to choose.

Acinar Cells ; chemistry ; Animals ; Calcium ; analysis ; Calcium Signaling ; Cells, Cultured ; Mice ; Microscopy, Confocal ; methods ; Pancreas ; cytology

OBJECTIVETo study the effect of calcium-sensing receptor (CaSR) agonists and antagonists on the expression of CaSR in neonatal mice with persistent pulmonary hypertension (PPHN), and to clarify the role of CaSR in neonatal mice with PPHN.

METHODSForty-nine neonatal mice were randomly divided into four groups: control (n=10), hypoxia (PPHN; n=11), agonist (n=13), and antagonist (n=15). The mice in the PPHN, agonist, and antagonist groups were exposed to an oxygen concentration of 12%, and those in the control group were exposed to the air. The mice in the agonist and antagonist groups were intraperitoneally injected with gadolinium chloride (16 mg/kg) and NPS2390 (1 mg/kg) respectively once daily. Those in the PPHN and the control groups were given normal saline daily. All the mice were treated for 14 consecutive days. Hematoxylin and eosin staining and immunohistochemistry were used to observe the changes in pulmonary vessels. Laser confocal microscopy was used to observe the site of CaSR expression and measure its content in lung tissues. qRT-PCR and Western blot were used to measure the mRNA and protein expression of CaSR in lung tissues.

RESULTSCompared with the control group, the PPHN group had significant increases in the pulmonary small artery wall thickness and the ratio of right to left ventricular wall thickness (P<0.05), which suggested that the model was successfully prepared. Compared with the control group, the PPHN group had a significant increase in the mRNA and protein expression of CaSR (P<0.05), and the agonist group had a significantly greater increase (P<0.05); the antagonist group had a significant reduction in the mRNA and protein expression of CaSR (P<0.05).

CONCLUSIONSCaSR may play an important role in the development of PPHN induced by hypoxia in neonatal mice.

Animals ; Hypoxia ; complications ; Lung ; pathology ; Mice ; Myocardium ; pathology ; Persistent Fetal Circulation Syndrome ; etiology ; pathology ; Pulmonary Artery ; pathology ; RNA, Messenger ; analysis ; Receptors, Calcium-Sensing ; analysis ; genetics ; physiology