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Background Corneal blindness is one of the major blinding eye diseases in China.With the development and progress of tissue engineering technology,tissue-engineered corneas offers a new approach to the treatment of corneal diseases.To select and cultivate ideal seed cells is a foundation of construction of tissueengineered corneas.Objective This study was to evaluate the efficiency of stripe off the Descemet membrane with endothelium plus enzymic digestion in the isolation of corneal endothelial cells and analyze the bionomics of rabbit corneal endothelial cells (CECs) in vitro.Methods Descemet membrane was stripped from fresh cornea of New Zealand rabbit under the dissection microscope.Descemet membrane with endothelium was incubated in trypsin and EDTA solution at 37 ℃ and then purified for CECs subculture in vitro.The morphology of the cultured cells was observed under the inverted microscope and marked by CM-Dil dye solution.Then the shape of the cells was observed by hematoxylin and eosin staining and the cells were identified for the expression of neuron specific enolase (NSE) using immunochemistry.The viability of the cells were evaluated by trypan blue staining.The surface structure of the cells were examined under the scanning electron microscope.Intercellular zonula occludens-1 (ZO-1) was identified by immunofluorecsence staining.Results A large number of purified CECs were obtained from Descemet membrane with endothelium through enzymic digestion.Cultured cells grew well and formed monolayer 5-7 days later with the cobblestone stone-like arrangement.The survival rate of the cells was 95％.CECs presented with the red annular fluorescence for CM-Dil with the labeling rate ＞90％.NSE was positively expressed in the cytoplasm.Polygon CECs were seen by hematoxylin and eosin staining and showed the brown staining.Abundant microvilli on the cellular surface and interconnected foot process were seen under the scanning electron microscope.ZO-1 showed the green fluorescence.Conclusions The method of striping off the corneal Descemet membrane with endothelium plus enzymic digestion can obtain abundant CECs.Cultured cells have good biological properties.This study may offer a feasible application in the engineering of corneal transplant membrane.
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Background Interleukin (IL)-1a-889C/T,IL-1β-S11C/T,+ 3962C/T and IL-1Ra-2 are different morphology of single nucleotide of interleuki-1 (IL-1).The potential relationship of IL-1 polymorphisms with Beh(c)et disease has been studied in several populations and groups.However,study outcomes are controversial for a long duration.Objective This Meta-analysis was to determine whether the IL-1 gene polymorphisms is associated with the pathogenesis and development of Beh(c)et disease.Methods The literature of the assoiation between IL-1 polymorphisms and Beh(c)et disease was retrieved from Medline,EMBASE,Cochrane Library,Web of knowledge,Google Scholar,Wanfang and CNKI databases.All the published original articles (case-control studies) were reviewed with the deadline May 31,2013,and the languages were limited to English and Chinese.Included reports were evaluated based on Newcastle-Ottawa Scale (NOS) score.The potential influence of IL-1α-889TT gene,IL-1β-3962C,IL-1β-511T and IL-1Ra-2 polymorphisms on Beh(c)et disease were analyzed.RevMan5.0,the Cochrane collaboration software program,was used to prepare and complete this review.The effect size was assessed using fixed effect model in the index with lower heterogeneity (I2 ＜50％) or randomized effect model in the index with higher heterogeneity (I2＞50％).Results The literature search resulted a total of 370 cases-controlled studies,and 7 studies met the included criteria,with the NOS scores ≥ 8.The total patients were 499 and normal controls were 708.Meta-analysis was performed on several populations.Overall,the polymorphisms of IL-1β-3962C allele increased the susciptibility of Beh(c)et disease (OR =1.41,95％ CI:1.06-1.88,P =0.02),and IL-1α-889TT genotype reduced the risk of Beh(c)et disease (OR =0.61,95 ％ CI:0.40-0.92,P =0.02).However,There was no significant association between variants of IL-1β-511T allele (OR =0.84,95％ CI:0.58-1.23,P=0.38) or IL-1RA-2 allele (OR =1.25,95％ CI:0.50-3.14,P=0.63) and Beh(c)et disease susciptibility.Conclusions These results suggest that Beh(c)et disease is associated with the IL-1 gene polymorphisms at the locations α-889C/T and β-3962C/T.A larger sample size clinical data still are need to confirm this conclusion.
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[ ABSTRACT] AIM:To evaluate the effects of 1α, 25-dihydroxyvitamin D3 on T helper cell 17 ( Th17 cells) and its related cytokines in a mouse model of corneal allograft transplantation.METHODS:C57BL/6 mice were transplanted with corneal grafts from BALB/c mice and treated intraperitoneally with 1.0μg 1α, 25-dihydroxyvitamin D3 or soybean oil every other day after operation.The transparency of the corneal grafts was evaluated for potential rejection signs by slit lamp biomicroscopy and histopathology.The expression levels of IL-17, RORγt and IFN-γin the spleen were measured by real-time PCR.Moreover, the protein expression of RORγt and IL-17 in the peripheral blood was analyzed by Western blotting. IL-17 and IFN-γin peripheral blood were measured by flow cytometry.RESULTS:1α, 25-dihydroxyvitamin D3 signifi-cantly inhibited the rejection of the corneal allograft and reduced the numbers of inflammatory infiltrates in the corneal graft. In the spleen, 1α, 25-dihydroxyvitamin D3 treatment reduced the expression levels of IL-17, RORγt and IFN-γ.In the pe-ripheral blood, 1α, 25-dihydroxyvitamin D3 treatment downregulated the expression levels of RORγt, IL-17 and IFN-γ. CONCLUSION:The effects of 1α, 25-dihydroxyvitamin D3 on suppressing corneal transplantation-induced allograft rejec-tion in mice are closely associated with its modulation on IL-17 and related cytokine RORγt.
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Objective To analyze the risk factors of chronic kidney disease complicated with acute kidney injury and its treatment efficacy.Methods 180 chronic kidney disease patients complicated with acute kidney injury were divided into the elderly group and the young group.The underlying diseases,risk factors of acute kidney injury and clinical efficacy were compared between the two groups.Results The proportions of chronic glomerulonephritis,nephrotic syndrome and lupus nephritis in young and middle-aged group were 37.8％,15.6％ and 20.0％,which were significantly higher than those in the elderly group (17.8％,8.9％,2.2％) (P ＜ 0.05).The proportions of diabetic nephropathy and hypertensive renal injury in elderly patients were 31.1％ and 24.4％,which were significantly higher than those in the young and middle-aged group (6.7％,4.4％) (P ＜ 0.05).In the old age group,the proportions of infections and renal toxicity drugs were 28.9％ and 22.2％,which were significantly higher than those in the young group.In the young and middle-aged group,the proportions of malignant hypertension and deterioration of the primary disease were 22.2％ and 26.7％,which were significantly higher than those in the elderly group (P ＜0.05).The total effective rate of the young group was 88.9％,which was significantly higher than 71.1％ of the elderly group (x2 =6.846,P ＜ 0.05).Conclusion Elderly and young and middle-aged chronic kidney disease patients complicated with acute kidney injury have differences in risk factors and clinical efficacy.
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Objective To study the effect of mycophenolate mofetil(MMF) on primary nephrotic syndrome and the impact on urinary protein.Methods According to the digital table,136 cases of nephrotic syndrome were randomly divided into MMF group and control group,68 cases in the control group were dealed with conventional treatment,MMF group was treated with MMF.After 6 months,the treatment effects of both groups were evaluated.Results The overall response rate of MMF group was 82.8％,higher than that of control group 60.6％ with statistically significant difference (x2 =10.86,P ＜ 0.05),;Blood urea nitrogen(BUN),serum creatinine(Scr),24 h urinary protein of two groups had varying degree of reduction,the indexes of MMF group decreased even more significant (t =2.50,4.59,11.12,all P ＜ 0.05).Serum albumin of the two groups had risen,but MMF group increased more significantly (t =3.09,P ＜ 0.05).Conclusion The advantages of MMF are significant effect,reduction in side effects,safe and reliable on nephrotic syndrome and urinary protein,which is worthy of clinical use.
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AIM: To provide a basis for appraising quality standard of Baohe Pill (Fructus Gataegi, Massa medicata fermentata, Rhizoma pinelliae, Pericapium citr reticulatae, Fructus forsythiae, Semen raphani, Fructus Mordei germinatus, Poria) inclusive of its RP-HPLC fingerprint chromatogram. METHODS: The chromatographic column was Kromasil C_ 18 column (4.6 mm?250 mm, i.d, 5-?m particle size). The mobile phase was 0.5% solution of ammonium dihydrogen phosphate (NH_4H_2PO_4), and the flow rate was 0.8 mL/min with UV detector at 214 nm. RESULTS: The RP-HPLC fingerprint chromatogram of Baohe Pill was established. In the experiment, for precision and repeatability, the RSD of each area of common peak was less than 3%. CONCLUSION: This method is simple and reliable.