Background Recombinant hirudin variant Ⅲ(rHV3) can effectively prevent galactose-induced human lens epithelial cells LECs injury,but little is known about the molecular mechanism of its action.Objective The present study was to investigate the effects of rHV3 on the expression of apoptosis-related genes in damaged LECs induced by galactose.Methods The rHV3 was extracted by our research group,and the biological activity of rHV3 was identified by titration of thrombase according to Markwardt's method.Human LECs (SRA01/04) were cultured using 125×10-3 mol/L D-galactose+10% FBS+D/F12 medium to establish the damaged human LECs model.rHV3 was added into the medium of the damaged human LECs model.Human LECs were cultured in D/F12 medium containing 10% FBS as normal control.The expression of apoptosis-related genes,such as aldose reductase (AR),bax,bcl2 and p53,in LECs at the mRNA level was detected using RT-PCR.The abundance ratio of target genes was presented with the absorbance (A) of gene mRNA/GAPDH mRNA.Results Compared to the normal control group,the A values of AR mRNA/GAPDH mRNA,bax mRNA/GAPDH mRNA and p53 mRNA/GAPDH mRNA were significantly elevated in model group (t=3.90E-06,t=8.44E-04,t=5.15E-08,P＜0.01).However,in the rHV3-treated group,the A values of AR mRNA/GAPDH mRNA,bax mRNA/GAPDH mRNA and p53 mRNA/GAPDH mRNA were lower than those of model group (t=5.90E-06,t=1.51E-04,t=3.42E-06,P＜0.01).The bcl2 mRNA/GAPDH mRNA was markedly downregulated in the model group when compared with the normal control group (t=1.86E-05,P＜0.01);while after rHV3 addition,bcl2 mRNA/GAPDH mRNA increased in comparison with the model group (t=8.56E-05,P＜0.01).Conclusion 125×10-3mol/L D-galactose induces the damage and apoptosis of human LECs.rHV3 likely plays a protective function on D-galactose-induced damage of human LECs by inhibiting the polyol pathway and mitochondria-mediated pathway.

AIM:To investigate the efficacy of controlled continuous curvilinear capsulorhexis(CCC) technique in short axial length and shallow anterior chamber eyes.METHODS:Sixty-eight patients(68 eyes) with short axial length and shallow anterior chamber were included.The routine CCC technique was used in 32 cases (32 eyes) and controlled CCC technique was used in 36 cases (36 eyes).The success rate and complication were compared between two groups. RESULTS:The success rate of the routine technique group and controlled technique group was 53. 13% and 86.11% respectively. Incomplete CCC leading to posterior capsule tears was 9.38% and zero in two groups respectively.CONCLUSION: Controlled CCC technique can increase the success rate and reduce complications in short axial length and shallow anterior chamber eyes.KEYWORDS:phacoemulsification; continuous curvilinear capsulorhexis; complication

Objective To determine the iodine content of common foods in different geographical areas (coastal city,coastal rural area,mountainous city and rural area) in Fujian Province and provide basic data for evaluation of dietary iodine intake.Methods In 2010,based on the types of food of the total diet study,one food sample(consumption rate is greater than 1％) was collected in coastal city(Taijiang),coastal rural area(Xiang'an),mountainous city (Sanyuan) and rural area(Mingxi).These foods including cereal,beans,potato,meat,eggs,milk,aquatic products,vegetables,fruits,sugar,beverages,liquor and seasoning,and so on 184 kinds of common foods.The iodine content of these food was tested by As (Ⅲ)-Ce4+ catalytic spectrophotometry.Results Among the 184 kinds of food tested,164 were not indicated food iodine content in the Chinese Food Composition Tables (2004).The iodine content of common food in Fujian Province in descending order were salt (30 000 μg/kg),aquatic products(341.4 μg/kg),eggs(255.9 μg/kg),dairy(106.7 μg/kg),meat(103.2 μg/kg),cereals (40.7 μg/kg),vegetables(30.7 μg/kg),beans(12.9 μg/kg),sugar(8.5 μg/kg),fruits(6.7 μg/kg),potatoes(2.4 μg/kg) and alcohol(2.1 μg/kg).The iodine content of kelp and laver was the highest in all the food,which was 314 780.1,176 956.5 μg/kg,respectively.The iodine content of food from animal(241.4 μg/kg) was much higher than that of the food from plant(25.4 μg/kg).The iodine content of common cereals,potatoes,beans,sea algae,meat,eggs and aquatic products was compared in the four areas,and the difference was not statistically significant (M =135.5,20.0,42.0,16.0,54.0,4.0,x2 =1.4,all P ＞ 0.05).The iodine content of vegetables and fruits was compared,and the difference was statistically significant (x2 =8.5,M =204.5,all P ＜ 0.05).Conclusion The iodine content of different foods is different.

To breed a new yam cultivar of Dioscorea alata, the different and excellent germplasm resources were investigated within artificially cultivated population and some superior individuals, with a higher yield and medicinal properties, were selected. Considering results of the yield and medicinal properties during 2006-2013 cropping season, strains and lines were established and selected. As a result, the yield of the new developed cultivar (Wenshanyao No. 1, WSY01-1) reached 2217. 0 kg per 667 m2 (fresh weight) and 348.3 kg per 667 m2 (dry weight), and increased 23.8% and 23.9% comparing with control cultivars (landraces). Comparing with control cultivars, the level of polysaccharide, allantoin, and dioscin increased 36.9%, 48.3%, 20.9%, and reached 12.2%, 1.30%, 579.7 µg · g(-1), respectively. This result showed that the systematic selection method can significantly improve yield and medicinal properties of D. alata, and the developed " Wenshanyao No. 1" exhibits wide spreading prospects.
Allantoin ; analysis ; Breeding ; Dioscorea ; chemistry ; genetics ; growth & development ; Diosgenin ; analogs & derivatives ; analysis ; Polysaccharides ; analysis

Objective To investigate the current status of coal-burning endemic fluorosis and the fluoride content in foods and drinking water in Hongya County,Sichuan Province.Methods Dental fluorosis and urinary fluoride were suveyed in children of 8～12 years old in two schools which repectively located in Gaomiao and Wawushan Town.The adults above 20 years old underwent clinical examination.At the same time,fifty adults above 20 years old in Garden Village were chosen to take forearm and calf X-ray films to find out the evidence of skeletal fluorosis.The content of fluoride in food such as bacon,corn,dry capsicum in Sanxing Village in Gaomiao Town and Futian Village in Wawushan Town as well as drinking water in five families in Sanxing Village were determined.Results The dental fluorosis rate of children was 40.76%(161/395),the dental fluorosis index was 0.86 in Gaomiao Town.The dental fluorosis rate of children was 14.36%(82/571),the dental fluorosis index was 0.31 in Wawushan Town.The medium value of the urine fluoride was 0.81 mg/L.ranged 0.16～3.89 mg/L.The positive rate oi the clinical examination of skeletal fluorosis was 5.27%(27/512),the X-rays detective rate was 4.00%(2/50).The medium value in bacon,corn,dry capsicum were 6.00,0.64,1.49 mg/kg.The averaged content of the fluoride in drinking water(0.14±0.06)mg/L of local household was within the eligible limitation.Conclusions It is currently a mild endemic disease in Hongya Country,its incidence is reduced apparently,pathogenetic environmental fluoride content is reduced.The main source of fluoride is from the preserved ham contaminated with fluoride,which is epidemiologically significant in endemic area of Hongya County.Defluoriding countermeasures should be taken in the endemic areas.

Objective To investigate the role of contrast enhanced ultrasound (CEUS) in evaluating and guiding radiofrequency ablation (RFA) of hepatocellular carcinoma (HCC) and its feeding vessels. Methods From January 2006 to June 2007, 71 patients with 75 hypervascular HCC in Peking University Cancer Hospital who underwent RFA were included in the study. The diagnosis was conifrmed by ultrasound guided biopsy for all patients. These patients were not suitable for transcatheter arterial chemoembolization (TACE) or had poor responds to TACE. They were divided into two groups, which included group percutaneous artery ablation (PAA) combining RFA and group RFA. There were 38 patients with 39 HCC in group PAA combining RFA and CEUS were used to identify the range of HCC inifltration. Firstly, PAA of the feeding vessels was conducted under the guidance of color doplor lfow imaging (CDFI). Then CEUS was performed to evaluate HCC perfusion after blocking the feeding vessels. Finally, the rest of the tumor was ablated by RFA. In group RFA, there were 33 patients with 36 HCC, who did not undertake PAA before RFA. Generally, the RFA was planned based on tumor size and location, and the ablation started with deep part of HCC or portion close to nearby organs. Contrast CT was used as a post-RFA imaging for follow-up at 1, 3 and 6 months post-RFA. T test was used to compare the difference in focal lesions number between two groups, andχ2 tests were used to compare the difference in necrosis rate between two groups after treatment. Results In group PAA combining RFA, post-PAA CEUS showed intratumor perfusion decreased more than 70%in 31 HCC (79.5%, 31/39). Of them, 13 HCC (33.3%, 13/39) showed complete perfusion defect with clear margin, called“solar eclipse sign”. The rest 8 HCC (20.5%, 8/39) showed 40%-70%of perfusion defect. In group PAA combining RFA, CDFI showed 35 (83.3%, 35/42) feeding vessels were blocked, and 3 vessels (7.1%, 3/42) showed signiifcant decreased lfow signal after PAA. There were average 3.18±1.42 ablations per HCC in group PAA combining RFA, and 4.32±1.56 in group RFA. The number of ablations per HCC in group PAA combining RFA was signiifcantly less than group RFA (t=2.524, P=0.015). The tumor necrosis rate at 1 month post-RFA in group PAA (92.3%, 36/39) combining RFA was signiifcantly higher than that of group RFA (66.7%, 24/35) (χ2=8.264, P=0.001). Conclusions With CEUS, PAA can effectively block the feeding vessels of HCC, enhance ablated necrosis in the tumor and signiifcantly increase necrosis rate post-RFA for large hypervascular HCC. CEUS-assisted PAA can improve efifciency of RFA with less ablation number and better result.

objective To explore the role of cytokines and dendritic cells (DCs) in rat high-risk corneal allograft survival pro- longed by superantigen Staphylococcal Enterotoxin B (SEB) and to compare the different effects between SEB and glucocorticoid. Design Experimental study.Participants Fisher 344 and Lewis rats.Methods The Fisher 344 and Lewis inbred rats were used for high-risk penetrating keratoplasty model.All of the Lewis rat recipients were divided into three groups by blinded fashion.GroupⅠand GroupⅡrats were injected intraperitoneally with 0.2ml saline buffer or SEB (75?g/ml) respectively at 4-day intervals on three occasions before transplantation.GroupⅢrats were injected subconjunctivally with 0.1ml dexamethasone (1mg/ml) daily from the first day after surgery for 2 weeks.The allograft survival was examined under slit-lamp.The concentration of interleukin IL-1?,IL-2,IL-4,IL-5,IL-6, IL-10,TNF-?in rat aqueous humor and peripheral blood were measured by liquichip and the cytokine and CD11c,CD80,MHC-Ⅱex- pression in corneal grafts were examined by immunohistochemestry staining.Main Octcome Measures Mean survival time of the allo- grafts,the level of cytokines in aqueous humor,peripheral blood and corneal grafts.Results Compared with group control,the grafts mean survival time was delayed about 3.8d in group SEB (P=0.00) and 7.1d in group dexamethasone(P=0.01).But there was no signifi- cant difference between group SEB and dexamethone (P=0.26).Liquichip test showed that the level of IL-1?in aqueous humor was re- duced and IL-4,IL-6 and IFN-?,ascended.Only IFN-?,and IL-6 could be found in peripheral blood,and the changing shift of them was similar to that in aqueous humor.The immunohistochemistry staining showed that the expression of IL-2 in rat corneal grafts was signif- icantly decreased but IL-4,IL-6 and IL-10 elevated.The expression of DCs in group SEB was similar to that in group control,which was elevated after keratoplasty,but the phenotype of DCs was not the same.There were predominantly mature DCs in group control while immature DCs in group SEB.Conclusions The immunological inhibiting effect of SEB is same to glucocorticoid,but the mecha- nism is different.SEB can modulate immune response,which might induce immune inhibition via the local production of cytokines and effect on DCs maturation involving corneal graft rejection prevention.

BackgroundPterygia is a clinical common disease.A lot of surgical methods are developed to decrease the recurrence rate.Resent years,the application of fibrin glue is receiving more and more attention.Objective This study was to explore the effects of fibrin glue in decreasing inflammatory irritation and its mechanism. Methods Pterygia models were created in 12 clean rabbits by exsection of limbal tissue and topical administration of 1.25％ diluted hydrochloric acid,and then the conjunctival autograft surgery was performed in the experimental rabbits.The conjunctival flap was sutured in the left eyes,and the conjunctival wound was closed using fibrin glue in the right eyes.The operation duration for each group was documented and compared.The irritation sign was examined under the slit lamp in all the rabbits 1 week and 4 weeks respectively.The expressions of vascular endothelial growth factor (VFGF) and basic fibroblast growth factor (bFGF) proteins in the conjunctiva tissue were detected by immunochemistry,and the expressions of VFGF mRNA and bFGF mRNA in the conjunctival tissue were determined by reverse transcription polymerase chain reaction (RT-PCR).Results The operative duration was (21.3±0.2) minutes in suture group and( 10.1 ±0.1 )minutes in the fibrin glue group with a significant difference between two groups( t =102.242,P＜0.05 ).From 1 week through 4 weeks,the hyperemiain degree was obviously slight in fibrin glue group compared with suture group.Immunochemistry showed that VEGF and bFGF proteins were expressed mainly in the cytoplasm of conjunctival epithelium layer.The positive response intensity was weaker in the fibrin glue group than in suture group 1 week and 4 weeks after operation.RT-PCR revealed that the expression level of VEGF mRNA was significantly lower in fibrin glue group than in suture group,and the VEGF mRNA was gradually decreased with the time lapse ( Fgroup =174.443,P =0.000 ; Ftime =231.459,P =0.000 ).The similar outcomes were found in the expression of bFGF mRNA(Fgroup=41.727,P=0.000;Ftime=55.417,P=0.000). ConclusionsThe use of fibringluecanshortentheoperationdurationandreducepostoperationinflammatoryreaction.The downregulation of VEGF and bFGF in tissue is the possible mechanism of remitting irritation sign,which allows a reduce of the recurrence rate of pterygia.

Carcinoma, Hepatocellular ; genetics ; metabolism ; DNA Methylation ; DNA Restriction Enzymes ; metabolism ; DNA, Neoplasm ; genetics ; Glutathione S-Transferase pi ; genetics ; metabolism ; Humans ; Liver Neoplasms ; genetics ; metabolism ; Polymerase Chain Reaction ; methods

OBJECTIVETo observe the expression profiles of osteoblast-related genes in human mesenchymal stem cells (MSCs) derived from bone marrow during osteogenic differentiation.

METHODSMSCs were induced to differentiate with MSC osteogenic differentiation medium for 7, 14, 21 and 28 days respectively. Alizarin Red staining was used to detect matrix mineralization. Expression of osteoblast-related genes, including osteocalcin, osteopontin, Runt-related transcription factor 2 (Runx2), alkaline phosphatase and collagen type 1, was assessed with quantitative reverse transcription-polymerase chain reaction.

RESULTSOn day 14 after induction of differentiation, cells were stained positively with Alizarin Red. The expression levels of these genes exhibited an upward trend as induction time was prolonged. Exposure to osteogenic differentiation medium less than 21 days did not significantly induce osteocalcin expression; osteocalcin expression levels in the differentiated cells induced for 21 and 28 days were 1.63 and 2.46 times as high as the undifferentiated cells respectively (all P<0.05). Stimulation with MSC osteogenic differentiation medium over 14 days significantly enhanced bone marrow-derived MSCs to express osteopontin and Runx2 genes (all P<0.05). Osteogenic differentiation medium could significantly induce the expressions of alkaline phosphatase and collagen type1 genes (all P<0.05). Their expressions reached the peak levels on day 21, which were increased more than 4- and 3-fold respectively.

CONCLUSIONHuman bone marrow-derived MSCs could exhibit the sequential expression pattern of osteoblast marker genes during osteogenic differentiation in vitro.

Alkaline Phosphatase ; genetics ; Cell Differentiation ; Cells, Cultured ; Collagen Type I ; genetics ; Core Binding Factor Alpha 1 Subunit ; genetics ; Genetic Markers ; Humans ; Mesenchymal Stromal Cells ; metabolism ; Osteocalcin ; genetics ; Osteogenesis ; Transcriptome