Background Oxidative damage is a major cause of age-related cataracts,and the ubiquitinproteasome system is involved in lens differentiation and development.Ubiquitin carboxy-terminal hydrolase L1 (UCHL1),one of key enzymes of ubiquitin-proteasome system,was discovered to participate in the age related diseases and oxidative stress damage. Objective This study was to investigate the effects of UCHL1 on the formation and development of age-related cataract. Methods Lens capsule were collected from 24 patients with age-related cataract(including 12 cases of cortical cataract and 12 cases of nuclear cataract) during the surgery.Five normal lens capsule membranes were obtained from eye bank of Tongji University.Human lens epithelial cells (LECs) line (SRA01/04) was also collected in this study.Expression of UCHL1 in the lens epithelial layer of different samples was assayed using immunofluorescence technology.UCHL1 eukaryotic expressing vector was constructed and transfected into cultured SRA01/04 by liposome,and green fluorescent protein (GFP) eukaryotic expressing vector was transfected at the same method as the control group.UCHL1 over-expressing cells were then exposed to different concentrations (0.2,0.3,0.4 and 0.5 mol/L) of tert-butyl hydroperoxide (TBHP) for 24 hours and subsequently monitored for cell viability evaluation by MTT assay. Results Immunofluorescence showed that UCHL1 was expressed in human lens epithelial layer,but significantly different expressing levels were seen among normal lens capsular membrane,cortical cataract and nuclear cataract ( F =13.411,P =0.000),and UCHL1 expressing levels were lower in cortical cataract and nuclear cataract than the normal lens (P =0.000,P =0.000).No significant difference was found in UCHL1 expressing level between cortical cataract and nuclear cataract ( P =0.164).Western blot analysis verified that UCHL1 exhibited a stranger expression in the UCHL1 transfected group compared with the GFP transfected group,illuminating a successful transfection of UCHL1 in SRA01/04 cells.MMT assay revealed that the A570/630 value in UCHL1 transfected cells was significantly elevated in comparison with GFP transfected cells following the treatment of 0.3 mol/L TBHP. Conclusions UCHL1 has an antioxidative ability,and it might plays an important role in the progress of age-related cataract.

Objective:To study the effects of PRF and recombinant hPDGF-AB,TGF-β1 and VEGF on the proliferation and adhe-sion of rat adipose tissue-derived stem cells(ADSCs)in vitro.Methods:ADSCs were cultured with PRF membrane and various do-ses of PDGF-AB,TGF-β1 and VEGF,cell adhesion was examined by adhesion assay after 2 culture,cell proliferation was examined by CCK-8 kit after 1 -7 d culture.Results:Cell adhesion assay showed that the adhesive numbers of rat ADSCs in PRF group were significantly higher than those in the negative group(P <0.05).The adhesive numbers of the ADSCs treated by PDGF-AB showed no significantly difference among different concentration groups(P >0.05).The adhesive numbers of the ADSCs treated by VEGF or TGF-β1 at different concentrations showed significant difference(P <0.05).CCK-8 kit assay showed that at different time points, the A values of ADSCs in PRF group were significantly higher than those of the negative control group(P <0.05).The A values of ADSCs in VEGF or PDGF-AB groups at different concentrations showed significant difference(P <0.05).The A values of rat AD-SCs in TGF-β1 group at different concentrations were lower than those in the negative control group(P <0.05).Conclusion:PRF as a combination of growth factors may stimulate the proliferation and adhesion of rat ADSCs in vitro.PDGF-AB and VEGF may stim-ulate the proliferation of rat ADSCs.TGF-β1 and VEGF may stimulate the adhesion of rat ADSCs in a dose-response manner to some degree.

Objective To treat cerebrovascular stenosis with percutaneous transluminal angioplasty and stenting and analyze the problems of this method and its therapeutic effects.Methods Twenty-three patients with brain watershed infarction proven by CT or MRI because of severe cerebrovascular stenosis were investigated for their clinic features,cerebrovascular digital subtraction angiography(DSA) and neurologic status before operation and after stent placement.Results The offending arteries concerning carotid sinus(C1 segment) in 14 cases,internal carotid artery(C2 segment) in 2 cases,internal carotid artery(C5,6 segment) in 4 cases;The offending arteries involving left artery in 8 cases,right in 15 cases.The percutaneous transluminal angioplasty and stenting for all patients was successful.All patients did not suffer from TIA and stroke in 6-to 12-month follow-up.Conclusion The brain watershed infarction caused by atherosclerotic cerebrovascular stenosis was not an infrequent ischemic cerebrovascular disease.The percutaneous transluminal angioplasty and stenting is an effective treatment for carotid artery or internal carotid artery stenosis.With proper selection of the patients and meticulous technique,percutaneous transluminal angioplasty and stenting should be of safety and efficacy for stroke prevention and treatment.

Objective To observe blood plasma matrix metalloproteinase-9 (MMP-9) of the patients with different types of carotid plaque and investigate the relationship between MMP-9 and carotid plaque vulnerability. Methods Totally 64 patients were examined by CDI, TCD, cranium CT, MRI, DSA and nerves function score (NIHSS) to judge their types of carotid plaque. Their plasma levels of MMP-9 and C-reactive protein (CRP) were detected. Results Plasma MMP-9 had direct correlation with plaque vulnerability(r=0.92,P

BACKGROUND:Cartilage injury is stil one of the clinical problems difficult to be treated completely so far. Recently, the discovery of synovial mesenchymal stem cells (SMSCs) has brought about the new hope to cartilage repair.
OBJECTIVE:To explore the process concerning SMSCs-based therapy for cartilage repair in the past few years, such as the characteristics of SMSCs, culture conditions, preclinical and clinical studies, and then to summarize the literatures published in recent years.
METHODS:A computed-based online search of PubMed and SpringerLink databases was performed using the key words of“synovial mesenchymal stem cells, cartilage repair”for literatures published from January 1993 to May 2013.
RESULTS AND CONCLUSION:Final y, 37 articles were included. SMSCs have a greater proliferative capability, colony-forming potential and chondrogenic potential than other mesenchymal stem cells. The diseases such as osteoarthritis and rheumatoid arthritis can influence the characteristics of SMSCs. Numerous articles have aimed at the studies of cellculture in vitro and celltransplantation in vivo. However, the process of SMSCs therapy is mostly at its preliminary stage. Reports on its unique characteristics, optimal culture conditions and the high-quality clinical studies are stil largely lacking. In a word, though further studies are needed, SMSCs appear to be a promising cellsource for cartilage repair in the future.

Objective To evaluate the clinical results of free vascularized fibular grafting (FVFG) for the treatment of osteonecrosis of the femoral head (ONFH).Methods From July,2009 to January,2013,85 cases (120 hips) of ONFH were treated with free vascularized fibular grafting.These cases included 61 males (87 hips) and 24 females (33 hips) with an average age of 36.5 years (22-51 years);7 hips (Ⅰ A 2 hips,Ⅰ B 3 hips,Ⅰ C 2 hips) were at stage Ⅰ,98 hips (Ⅱ A 24 hips,ⅡB 39 hips,ⅡC 35 hips) at stage Ⅱ and 15 hips (ⅢA 9 hips,ⅢB 4 hips,Ⅱ C 2 hips) at stage]Ⅲ according to the classification system of Association Research Circulation Osseous (ARCO).The mean preoperative Harris hip score was (60.21±6.85) points (42-71 points),The follow-up items included the X-ray examination,the Harris scores of the hip,and the evaluation of the complications.Results Eighty-three cases (117 hips) were followed up.The average duration of follow-up was 25 months (range from 12 months to 42 months).The mean postoperative Harris hip score was increased to (81.26±5.84) points (67-91 points) by the end of the follow-up,compared with the preoperation,the score improved significantly,the difference was statistically significant (P＜0.05).Comparing with postoperative X-ray,101 hips (86.3％) were improved,12 hips (10.3％) had no significant changes and deterioration occurred in 4 hips (3.4％).Conclusion The free vascularized fibular grafting is an effective method for treating osteonecrosis of and preventing the collapse of the femoral head.

To establish an HPLC method for determination of dactylorhin A and militarine in Cremastrae Pseudobulbus/Pleiones Pseudobulbus. The analysis was achieved on an Alltech Prevail C18 column (4. 6 mm x 250 mm, 5 microm) using a mobile phase of acetonitrile (A), water (B) gradient elution in a total run time of 35 min (0 min, 20:80; 30 min, 55:45; 35 min, 55:45) and a diode array detector was set at 224 nm. The flow rate was 0.8 mL x min(-1). The assay displayed good linearity over the concentration range of 0.257-9.95 microg (r = 0.999 8), and 0.128-10.27 microg (r = 0.999 9), respectively. The average recoveries (n = 9) were 94.70% and 102.8% for dactylorhin A and militarine, respectively. The method is accurate, quick, simple and reproducibility. It can be used for the quality control of Pleione bulbocodioides and Pleione yunnanensis.
Chromatography, High Pressure Liquid ; Glucosides ; analysis ; Orchidaceae ; chemistry ; Succinates ; analysis

Objective To investigate the ecological structure of intestinal fungal flora of patients with chronic hepatitis B infection. Methods HBV-liver cirrhosis patients, chronic hepatitis B patients, HBV carriers, and healthy volunteers were selected as research subjects to extract the total DNA from stool of each subject, 18S rRNA genes of intestinal fungi was cloned and sequence was analyzed for the construction of 18S rDNA clone libraries and phylogenetic tree, diversity and structural characteristics of intestinal fungal flora in each group was then analyzed. Results A total of 29 operational taxonomic units (OTUs) of intestinal fungal flora in all research subjects was acquired. All positive clones belonged to three fungal taxa: Zygomycetes (3.4%) , Ascomycetes (82. 8%) and Basidiomycetes (13. 8%) . The dominant fungal community was Candida spp. , uncultured fungus and Saccharomyces spp. , which accounted for 29. 2% , 15. 9%, 15.0% respectively. The OTUs which belonged to HBV-liver cirrhosis patients, chronic hepatitis B patients, HBV carriers, and healthy volunteers was 20,16,12,14 respectively. Conclusion There is an abundant fungal flora in the human intestine. Ecological structure of intestinal fungal flora appears to be various in patients with chronic hepatitis B infection, which indicate the close relationship between the alteration of ecological structure of fungal flora and the stage of chronic hepatitis B infection.

AIM: To explore the protective effect of curcumin on high glucose-induced decrease in contraction of isolated rat aortic rings, and to elucidate its underlying mechanism. METHODS: The thoracic aortic rings with endothelium of male Sprague-Dawley rats were mounted on a bath system. Isometric contractions of aortic rings were measured. HO activity was also evaluated. RESULTS: (1)Four hours after incubated with 44 mmol/L of glucose (high glucose),the vascular contraction responses to phenylephrine (PE) decreased compared to control group (containing 11 mmol/L of glucose). (2)Coincubation with curcumin (3×10~(-11)-3×10~(-10) mol/L) and high glucose,the high glucose-induced decrease in contraction responses to PE of arteries was partly inhibited. (3)Four hours after incubation with curcumin,the HO activity in thoracic aorta increased. ZnPP,an inhibitor of HO-1,completely abrogated the protection effect of curcumin. (4)Methylene blue,an inhibitor of guanylate cyclase (GC),partly abolished the protective effect of curcumin. CONCLUSION: Curcumin prevents the high glucose-induced decrease in contraction responses to PE in intact aortic rings. The mechanism might be mainly involved in the activation of HO-1 and GC.

Background The underlying mechanism of choroidal neovascularization(CNV) is multifactorial and complex.Focal adhesion kinase(FAK) plays a crucial role in controlling essential cellular processes and influencing distinct steps of the angiogenic response.But to our knowledge,seldom study on the effect of FAK on CNV formation has been reported previously.Objective In this study,the effect of several CNV risk factors on the expression of FAK in cultured retinal pigment epithelium(RPE) cells was investigated to illuminate effect of FAK on CNV.Methods Human RPE cells were isolated from donor eyes and exposed to H2O2,swallow of outer segment of photoreceptors(POS) and extracellular matrix(ECM) separately with the treating as follows:RPE cells were co-cultured with 10,20,50 and 100μmol/L H2O2 for 20 days;POS(1×106/ml) were co-cultivated with RPE cells for 20 days(setting control group,POS group,hypoxia group with 200μmol/L CoCl2,and POS+hyoxia group);RPE cells were cultured on the plates coated with 100mg/L fibronectin(FN),laminin(LN) or collagen typeⅠfor 30minutes or 1 hour.The expression of FAK and pFAK in RPE cells were examined by Western blot analysis.Results FAK was highly expressed in the 20μmol/L and 50μmol/L H2O2 groups compared with control group(P＜0.01);while he expression level of pFAK was reduced after treated with H2O2 in comparison with the control group(P＜0.01).After cultured with POS for 20 days,the undigested lysosome could be observed in RPE cells.The expressions of FAK and pFAK in RPE cells were not significantly changed between control group and POS groups(P＞0.05),but those in hypoxia group were significantly up-regulated in comparison with control group(P＜0.01).Compared with the hypoxia group,the expression amount of pFAK was elevated in POS+hyoxia group(P＜0.01).In comparison with control group,the increased pFAK expression was seen in FN,LN and collagen typeⅠtreating for 1-hour groups(P＜0.05,P＜0.01).Conclusion FAK pathway participates in several CNV-initiated signaling,such as H2O2,POS and ECM,in cultured RPE cells.It is reasonable to believe that FAK potentially plays an important role in CNV-dependent disorder.