Objective To provide strong evidence for sterilization standardization by comparing the results of bacteria culture after conjunctiva sac irrigation by gentamicin and povidone-iodine (PVI) and analyzing the sterilization effect of these two common disinfectant.Methods Six randomized control trials comparing the sterilization effect between gentamicin and PVI in conjunctiva sac irrigation were retrieved through the following databases:Pubmed,Embase,Cochrane Library,WANFANG DATA,CNKI and VIP database.Relative Risk (RR) was set as the effect indexes to evaluating the conjunctiva sac bacterial-culture results before and after irritation.The data of these articles were processed with Meta analysis by random model.The side effects of both dilutions were also compared using the descriptive analysis.Results Before irrigation,bacteria in the sac were mostly gram-positive coccus,among which Staphylococcus epidermidis and Staphylococcus aureus were most common.The detection rate of these two bacteria were 30.5％ (226/740) and 2.4％ (18/740),and the constituent ratio were 65.5％ and 5.2％ respectively.After irrigation,the number of the bacteria was decreased dramatically,but could not be totally eliminated.There was no significant difference in sterilization effect between gentamicin and PVI after conjunctiva sac irrigation.The weighted average value of RR was 0.51 with 95％ confidence interval (CI) in (0.25,1.06),P =0.07.Comparing to gentamicin,PVI was less comfortable after conjunctiva sac irrigation,but when the concentration was regulated,it would be acceptable by patients and didn't affect surgeries.Conclusions There was no significant difference in sterilization effect between gentamicin and PVI in conjunctiva sac irrigation.The side effect of PVI was minor in low concentration.PVI is more specific to the bacteria species than gentamicin.Using PVI was recommended in preoperation sterilization.

[ABSTRACT]AIM:TostudytheeffectofsmallinterferingRNA(siRNA)ontheexpressionofbeta2-microglo-bulin (β2M) in pre-differentiated bone marrow mesenchymal stem cells (BMSCs).METHODS: The β2M siRNA was transfected into the pre-differentiated BMSCs with Lipofectamine 2000.BMSCs were divided into transfection group , blank control group and negative control group .The expression of β2 M at mRNA and protein levels was determined by real-time qPCR, Western blotting and laser confocal microscopy .The productions of aggrecan and type II collagen in pre-differentia-ted BMSCs were determined by toluidine blue staining and type Ⅱcollagen immunofluorescence .RESULTS:The results of real-time qPCR, Western blotting and laser confocal microscopy showed that siRNA successfully inhibited the expression ofβ2 M at mRNA and protein levels in the pre-differentiated BMSCs .The results of toluidine blue and type Ⅱcollagen im-munofluorescence staining showed that siRNA does not affect the productions of aggrecan and type Ⅱ collagen in the pre-differentiated BMSCs .CONCLUSION:siRNA targeting β2 M reduces the expression of β2 M in the pre-differentiated BM-SCs and does not affect the chondrocyte characteristics of pre -differentiated BMSCs .

[ABSTRACT]AIM:Toinvestigatetheinhibitoryeffectsofresveratrolonchondrosarcomaandtherelationwith mitochondrial and PI3K/Akt pathways.METHODS:Chondrosarcoma SW1353 cells were treated with resveratrol at con-centrations of 25, 50 and 100 μmol/L for the time intervals of 24 h, 48 h and 72 h.The viability and apoptosis of the SW1353 cells in the presence or absence of resveratrol were analyzed by CCK 8 assay and Hoechst 33258 staining , respec-tively.The protein levels of Bcl-2, Bax, activated caspase-3, Akt and p-Akt were detected by Western blotting .The cell migration ability was determined by wound scratch assay .RESULTS:Exposure of the cells to resveratrol resulted in a de-crease in the cell viability in a dose-and time-dependent manner (P<0.05).visible nuclei with apoptotic characteristics in resveratrol group were observed .The protein levels of activated caspase-3 and Bax were increased , and Bcl-2 and p-Akt were decreased compared with control group .The total Akt were not significantly changed .Resveratrol also significantly re-duced the migration of tumor cells .CONCLUSION:Resveratrol induces apoptosis of chondrosarcoma , which plays a role of part through mitochondrial and PI 3K/Akt signaling pathways .

Objective To investigate the effects of sevoflurane pretreatment on renal ischemia-reperfusion (I/R)-induced apoptosis in kidney in rats. Methods Thirty pathogen-free male SD rats weighing 220-260 g were randomized into 3 groups (n=10 each):group control (group C);group I/R and group sevoflurane(group S). Renal I/R was induced by clamping the left renal pedicle for 45 min in I/R and S groups. In group S inhalation of 2.2% sevoflurane in O2 was started at 30 min before operation and maintained throughout the experiment.Venous blood samples were taken at 3 h of reperfusion for determination of serum BUN and Cr concentrations. The animals were then sacrificed and the left kidneys were removed for microscopic examination, detection of apoptosis(by TUNEL)and determination of heme oxygenase-1(HO-1) mRNA and protein expression (by RT-PCR and Western blot).Results Renal I/R significantly increased serum BUN and Cr concentrations, apoptotic index(percentage of apoptotic cells) and the severity of necrosis of renal proximal convoluted tubules (0=normal,4=necrosis of whole segment of proximal convoluted tubules).Sevoflurane inhalation attenuated the I/R-induced changes mentioned above.HO-1 mRNA and protein expression was up-regulated by I/R and HO-1 mRNA expression was further up-regulated by sevoflurane inhalation.Conclusion Sevoflurane pretreatment can protect kidney against I/R injury by attenuating cell apoptosis.Up-regulation of HO-1 mRNA expression may be involved in the mechanism.

BACKGROUND:Many in vivo and in vitro experiments indicate that hypoxic co-cultures promote stem cells differentiate into chondrocytes.
OBJECTIVE:To evaluate the influence of hypoxia on the chondrogenic differentiation of three-dimensional co-cultured adipose-derived stem cells and articular chondrocytes.
METHODS:Adipose-derived stem cells and articular chondrocytes were mixed at the ratio of 3:1, then the mixed cells were seeded onto poly(lactic-co-glycolic acid)-gelatin scaffold at the ultimate concentration of 5.0×1010/L. The cells were cultured in normoxia (20%O 2 ) and hypoxic (5%O 2 ) conditions for 6 weeks. After culture, hematoxylin and eosin staining was performed for histological structure analysis, and alcian blue staining was used to evaluate glycosaminoglycan synthesis. Type II col agen expression was detected by immunohistochemistry staining. The content of DNA, glycosaminoglycan and hydroxyproline in the scaffold-cellcomplex was measured.
RESULTS AND CONCLUSION:In the hypoxia group, hematoxylin-eosin staining showed the formation of massive cells and extracellular matrix;alcian blue staining showed massive glycosaminoglycan formation;immunohistochemistry staining detected strongly positive expression of col agen type II, the content of DNA, glycosaminoglycan and hydroxyproline was higher than the normoxia group. Hypoxia promotes in vitro chondrogenic differentiation of co-cultured adipose-derived stem cells and articular chondrocytes. .

Objective: To investigate the mRNA expression of chemokine receptors in human villi and trophoblasts of first trimester gestation . Methods: The authors first obtained villous tissues from fifteen women who had undergone selective termination at 5 - 10 weeks of normal gestation. Total RNA was then extracted, using the TRIzol reagent, from villous tissues or Percoll-gradient purified trophoblasts. Consequently, the expressions of chemokine receptors in villous tissues and trophoblasts were investigated by way of semi-quantitative reverse transcriptase-polymerase chain reaction.Results: The chemokine receptors, CXCR4 and CXCR6, were highly expressed in each villous tissue, while the CCR6, CCR7, XCR1 and CX3CR1 were moderately expressed in villi. The chemokine receptors, CCR1- CCR5, CCR8 - CCR10, CXCR1 -CXCR3, were expressed only in some villous samples, while no CXCR5 mRNA was found in any villous tissue. The authors also found that the freshly isolated and Percoll-purified trophoblasts expressed CCR1, CCR3 - CCR5, CCR8 - CCR9, CXCR1 - CXCR4, CXCR6, XCR1 and CX3CR1 mRNA. Conclusion: A variety of chemokine receptors were expressed in villous tissues and trophoblasts of human first trimester gestation, hence, these receptors may play an important biological role at the materno-fetal interface in normal human pregnancy.

AIM:Toinvestigatewhetherautophagyisup-regulatedwhenresveratrol(Res)inducesapoptosis in chondrosarcoma , and to study the effects of autophagy inhibitor combined with Res on chondrosarcoma .METHODS:SW1353 cells were divided into 4 groups: control group, Res group, 3-methyladenine (3MA) group, and Res +3MA group.Electron microscopy was used to observe the autophagyosomes in control group and Res group .At the same time, the viability of the cells in the 4 groups was detected by CCK-8 assay.TUNEL staining and Western blotting (for determi-ning the levels of cleaved caspase-3, Bax and Bcl-2) were used to reflect levels of apoptosis in all groups .The expression of autophagy-related proteins Beclin 1, LC3-Ⅱ and p62 was detected by Western blotting .RESULTS: Exposure of the cells to Res resulted in a decrease in cell viability and an increase in the level of apoptosis ( P<0.05 ) .Compared with control group, the level of apoptosis was increased but the autophagy was decreased (P <0.05).Compared with Res group, the cell viability and the level of autophagy were decreased and the level of apoptosis was increased ( P<0.05 ) . CONCLUSION:Resveratrol induces apoptosis and autophagy , and inhibition of autophgay enhances resveratrol-induced apoptosis in chondrosarcoma .

AIM:To investigate the effect of oleuropein on interleukin-1β( IL-1β)-induced SD rat articular chondrocytes .METHODS:The SD rat articular chondrocytes were isolated by 2 step enzyme digestions .The chondrocytes were cultured in vitro.Inverted microscopic observation was performed during the culture .Alcian blue staining and type II collagen immunohistochemical staining were used to identify the chondrocytes .The effects of oleuropein on the viability of chondrocytes were determined by CCK-8 assay.The cells in 3rd passage were pretreated with oleuropein at 10, 50 or 100 μmol/L and subsequently stimulated with IL-1βat 10 μg/L for 24 h.Production of prostaglandin E 2 ( PGE2 ) and ni-tric oxide (NO) were evaluated by the Griess reaction and an enzyme linked immunosorbent assay (ELISA).The mRNA expression of matrix metalloproteinase ( MMP)-1 and MMP-13 was measured by real-time PCR.The protein levels of in-ducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2) and nuclear factor-kappa B (NF-κB) were detected by Western blotting .RESULTS:The cell viability of chondrocytes was not significantly impaired by treating with oleuropein at concentration of 10, 50 or 100μmol/L for 24 h compared with control group .Pretreatment with oleuropein significantly in-hibited the production of PGE 2 and NO induced by IL-1β.Oleuropein also significantly decreased the IL-1β-stimulated MMP-1 and MMP-13 mRNA expression in articular chondrocytes .Pretreatment with oleuropein inhibited the IL-1β-media-ted activation of NF-κB by suppressing the degradation of its inhibitory protein IκBαin the cytoplasm .CONCLUSION:Oleuropein inhibits IL-1β-induced inflammatory gene expression by suppressing NF-κB activation at the transcriptional le-vel, suggesting a new mechanism for the anti-inflammatory effects of oleuropein as a novel agent on treating with osteoarthri-tis.

Background Researches showed that elevatory blood glucose level results in long-term damage of cells and tissue,or metabolic memory phenomenon,and manipulation of hyperglycemic memory is a good approach in the prevention of diabetic complications.However,its mechanism is not clear.It is speculated that the pathogenesis of diabetic retinopathy (DR) in diabetic patients may be associated to related mechanisms.Uncoupling proteins (UCPs) can decrease the production of reactive oxygen species (ROS),which may be related to DR.Objective This study was to explore the association between DR and the single nucleotide polymorphisms (SNPs) of UCP genes in Chinese Han population with type 2 diabetes.Methods A cross-sectional study was performed.This study was approved by Ethic Committee of Affiliated First Hospital of Shanghai Jiao Tong University and complied with Declaration of Helsinki,and written informed consent was obtained from each subject prior to any medical examination.One thousand eight hundreds and seventy-five patients with type 2 diabetes mellitus were enrolled in Xinjing district of Shanghai city by cluster sampling from November 2014 to January 2015.The demographic and medical baseline characteristics,ocular examination and laboratory tests were obtained and periphery blood of 2 ml was collected for extraction of DNA.Eight tag SNPs of UCP1,three tag SNPs of UCP2,and seven tag SNPs of UCP3 were selected as marker locus for the detection of genotype by Sequenom Mass ARRAY.Matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry platform were used for genotyping.Hardy-Weinberg equilibrium (HWE) analysis,allele and genotype frequencies,haplotype analysis,and association tests for DR and SNPs were performed by SAS and SHEsis software.Results A total of 530 DR patients were checked out from 1 875 subjects with type 2 diabetes mellitus,with the detection rate of 28.27％.rs660339 locn of UCP2 gene and rs1626521,rs668514 locus of UCP3 gene appeared to have low detectable rates,and the secondary allele base frequency of rs632862 in UCP2 gene was ＜0.01 and rs15763 of UCP3 gene was unmatched with HWE,therefore,these locus analysis was not included.In 13 SNPs locus included in the analysis,only 2 SNPs of UCP1 gene were related to DR.Compared with the non-diabetic retinopathy (NDR) patients,the G allele frequency of rs10011540 was increased (P =0.03,OR =1.31,95 ％ confidence interval[CI] =1.03-1.67,and T allele frequency of rs3811787 was decreased (P=0.04,OR=0.86,95％ CI=0.75-0.99) in DR patients.Genotyping detection showed that the C/C and A/A frequencies of rs3811790 in UCP1 gene were significantly more and C/A frequency was less in DR patients than those in NDR patients (all at P＜0.01).The logistic regression analysis indicated an association of SNPs of rs10011540 and rs3811787 with DR independent from glucose and disease duration.Conclusions The SNPs of rs10011540 and rs3811787 locus in UCP1 gene are associated with DR in Chinese type 2 diabetes patients.

Objective To investigate prevalence of metabolic syndrome (MS) in population at high-risk for diabetes in Changfeng community of Shanghai. Methods Totally, 982 persons aged 40-79 years without previous history of diabetes, but with risk factors for diabetes from Changfeng community in Shanghai were screened by questionnaire, physical examinations, lipid profile and oral glucose tolerance test. Prevalence of MS and its clinical characteristic were observed. Results Overall prevalence of MS was 44. 6% (438/982) in Changfeng community, significantly higher in women of 48.6% (341/701) than that in men of 34. 5% (97/281) (P<0.01). Prevalence of MS increased with age in women (P <0. 01 ), but not in men (P>0.05). Components of MS ranking from high to low were hypertension of 66. 5% (653/982), abdominal obesity of 62.8% (617/982), decreased high-density lipoprotein-cholesterol (HDL-C) of 49. 4% (485/982), increased triglyceride (TG) of 46. 0% (452/982) and increased fasting glucose of 35.7% (351/982). There were 55.9% (245/438) of MS patients with three or more MS components in addition to abdominal obesity. Conclusions Prevalence of MS is higher in community population at high-risk for diabetes and more than half of the MS patients have three or more risk factors for cardiovascular disease.