Objective To study the endogenous metabolites of liver-kidney deficiency syndrome in knee-joint osteoarthritis (KOA) and explore the metabolic profile of KOA liver-kidney deficiency syndrome. Methods Totally 50 cases of KOA with liver-kidney deficiency syndrome and 50 cases of KOA with non-liver-kidney deficiency syndrome were collected respectively, and 20 cases of healthy volunteers were collected as the normal group. The serum samples of subjects were collected after fasting for 8 h. Hydro-Nuclear Magnetic Resonance (1H-NMR) spectrometer was collected. Principal component analysis and partial least squares discrimination analysis were used to conduct multivariate statistical analysis. Results 1H-NMR could identify 23 kinds of metabolites, and there was statistical significance between KOA patients and healthy volunteers (P<0.05, P<0.01). There was statistical significance in cartilage matrix metabolism, energy metabolism, lipid metabolism, pain, inflammation-related metabolites in patients with KOA liver-kidney deficiency syndrome and patients with KOA non-liver-kidney deficiency syndrome (P<0.05). Conclusion Patients with KOA liver-kidney deficiency syndrome have a unique 1H-NMR metabolic profile, KOA syndrome has a metabolic material basis.

Objective To detect the COMP and aggrecan content in serum of patients with KOA and explore its correlative factors.Methods COMP and aggrecan in serum of 100 patients with knee-osteoarthritis(KOA group)and 30 healthy volunteers (control group)were measured by ELISA.Correlative factors for COMP and aggrecan levels were tested by linear regression analy-sis.Results COMP and aggrecan levels in KOA group were significantly higher than that in control group (P < 0.05 ).Serum COMP and aggrecan levels were positively correlated with age,body mass index(BMI),WOMAC scores and X-ray K-L grade(P <0.05).Conclusion Serum COMP and aggrecan are useful markers in the diagnosis,which their level in KOA are higher than the normal people.Moreover,these rise of levels positively correlated with clinical disease parameters and radiological joint damage.

Objective To explore the value of carbon nanoparticles in patients with papillary thyroid carcinoma (PTC) undergoing total thyroidectomy combined with ipsilateral central lymph node dissection.Methods 43 patients with unilateral PTC were retrospectively analyzed.All patients underwent total thyroidectomy combined with central lymph node dissection.Patients were divided into carbon nanoparticles group and the control group according to whether carbon nanoparticles were used in the operation.The operation time,postoperative hospitalization time,the serum calcium level and its rate of change,the parathyroid hormone and its rate of change on the 1 st day after surgery,the number of central lymph node and the transfer rate,and the postoperative complications were compared between the 2 groups.Results There was no statistical difference between the 2 groups in operation time(P ＞ 0.05),while the postoperative hospitalization time of the carbon nanoparticles group was shorter than that of the control group(P ＜ 0.05).The serum calcium and the parathyroid hormone on the 1 st day after surgery in the carbon nanoparticles group was (2.31 ± 0.13) mmol/L and (33.45 ± 14.37) pg/ml respectively,higher than those of the control group (P ＜ 0.05).The low blood calcium rate (3/20 (15％)),the temporary hypoparathyroidism rate (2/20 (10％)) and the decline degree of parathyroid hormone (47.3 ± 14.31)％ in the carbon nanoparticles group were lower than those of the control group(P ＜0.05).The number of central lymph node dissected (9.45 ± 2.33) pieces/case in the carbon nanoparticles group was more than that of the control group (P ＜ 0.05).The number of lymph node in the right recurrent laryngeal nerve (3.12 ± 0.65) pieces/case was more than that of the control group(P ＜ 0.05).The lymph node metastasis rate had no significant difference between the 2 groups(P ＞ 0.05).The incidence of postoperative complications in the carbon nanoparticles group was lower than that of the control group (P ＜ 0.05).Conclusion The application of carbon nanoparticles in total thyroidectomy combined with central lymph node dissection can contribute to the recognition and protection of parathyroid glands and its blood supply,improve the rate of central lymph node dissection,and reduce the incidence of postoperative complications.

Objective To explore the roles of MircroRNA-217 ( Mir-217 ) , silent information regulator 1 (Sirt1), and hypoxia-inducible factor-1α(HIF-1α)in high glucose-induced inflammation and fibrosis in rat glomerular mesangial cells( RMCs) . Methods RMCs were pre-incubated with a Sirt1 activator resveratrol prior to high glucose treatment or transfected with Sirt1 small interfering RNA( siRNA) , HIF-1αsiRNA, and Mir-217 inhibitor. Real-time PCR was used to analyze the expressions of Mir-217, Sirt1 mRNA, and HIF-1α mRNA; Western blot was used to observe the protein expressions of Sirt1, HIF-1α, connective tissue growth factor(CTGF), endothelin-1(ET-1), and fibronectin( FN) . Enzyme-linked immunosorbent assay was applied to detect protein expression of transforming growth factor-β1(TGF-β1)and vascular endothelial growth factor(VEGF). Results High glucose increased Mir-217, HIF-1α, CTGF, ET-1, FN, TGF-β1, and VEGF expressions(all P<0. 01), while decreased Sirt1 expression. In addition, Mir-217 gene silencing or 25μmol/L resveratrol suppressed high glucose-stimulated expressions of HIF-1α, CTGF, endothelin-1, FN, TGF-β1, and VEGF(all P<0. 01). Conclusion Mir-217 mediates high glucose-induced inflammation and fibrosis in RMCs via Sirt1/HIF-1αsignal pathway. This study provides new evidence to clarify the protective mechanisms of Sirt1 in diabetic nephropathy.

Objective To study the risk factors affecting neoplasm recurrence and survival of hepatocellular carcinoma(HCC)following liver transplantation(LT).Methods The clinicopatholo gic data,neoplasm recurrence and survival results of 118 patients with HCC receiving LT were retro- spectively analyzed and various clinicopathologic risk factors for neoplasm recurrence and survival were evaluated by univariate and multivariate analysis.Results The follow-up time ranged from 1 to 32 months.The recurrence rate was 37.3% and the mortality was 35.5%.The 12-,18-,24-month survival rate was 84.55%,70.30% and 62.24%,respectively.The 12-,18-,24-month neoplasm free survival rate was 69.05%,66.93% and 61.38%,respectively.In the univariate analysis,por- tal vein neoplasm thrombus(PVTT),Milan-criteria,neoplasm size,histological differentiation and pTNM stage were associated significantly with neoplasm recurrence,and PVTT,Milan-criteria,pre- operative alpha-fetoprotein(AFP),histological differentiation and pTNM stage were associated signif- icantly with survival rate;In the multivariate Logistic regression analysis,PVTT and histological dif- ferentiation were independent predictive factors of neoplasm recurrence,and multivariate Cox regres- sion analysis showed that PVTT and AFP independently associated with prognosis.Conclusions PVTT and histological differentiation are the most important predictive factors of neoplasm recur- rence,and PVTT and AFP independently predict the survival of patients undergoing LT.

Objective To evaluate the clinical efficacy ofDuhuo Jisheng Decoction in treating liver and kidney deficiency syndrome of knee osteoarthritis (KOA) by ultrasound.Methods A total of 120 cases of KOA patients with liver and kidney deficiency syndrome were randomly divided into two groups (each contains 60 cases). Control group took hydrochloric acid glucosamine capsules, while treatment group tookDuhuo Jisheng Decoction additionally for 8 weeks. ESR, CRP, WOMAC, joint swelling scores, and TCM syndrome scores of the two groups were determined, and the sonographic characteristics of knee-joint effusion in patellar fabricius, synovial membrane, cartilage and bone erosions were compared before and after treatment.Results There were significant differences in ESR, CRP, WOMAC, joint swelling scores, and TCM syndrome scores between before and after treatment in both groups (P<0.05,P<0.01). The knee-joint synovial membrane got thinner after therapy in osteoarthritis patients, and the effusion in knee-joint also decreased. The vessel flow signals in knee-joint synovial membrane of osteoarthritis decreased after drug therapy (P<0.05,P<0.01).ConclusionDuhuo Jisheng Decoction can improve clinical efficacy of KOA with liver and kidney deficiency syndrome. Ultrasound can clearly observe the changes of effusion in patellar fabricius, and synovial membrane, and can provide good imaging assessment for the drug therapy for KOA patients.

Background Efficient and lowcost way to isolate keratocytes is helpful for research on cornea.Either relatively expensive or inefficient is the shortage of those means now applied,while raising the keratocytes through passage will change the phenotype of them quickly.Our aim is to approach the way getting keratocytes effectively utilizing modified two step enzymatic digestion by type I collagenase. Objective To evaluate the effect of isolating the bovine keratocytes utilizing two step enzymatic digestion and observe the morphological changes of the keratocytes during cultivation in vitro. Methods Keratocytes were isolated from bovine corneas using 0.5 mg/mL and 1 mg/mL type I collagenase digestion.The harvesting rate and viability rate of the primary keratocytes were evaluated.During the primary cultivation in vitro,the morphological changes of the keratocytes and their F-action distribution were observed.Results(1)The extracellular matrix of the bovine corneas were almost dissolved by the two step enzymatic digestion,followed the keratocytes completely isolated from the solid matrix.The amount of the harvested keratocytes from each cornea was(2.11±0.15)X106 on average while the viability rate was(91.69±3.73)% and the inoculation rate Was(81.20±1.25)%.(2)The primary keratocytes attached and spreaded out with dendritic and stellate morphology.After 3 days cultured,the branches of the keratocytes were contacting and formed networks.The F-actin detected by phalloidin binding showed a limited cortical localization. Conclusion (1)The method of two step enzymatic digestion can make the extracellular matrix of bovine cornea stroma completely degraded with the advantages in high efficiency of harvesting keratocytes and high cell viability and relatively simple manipulation. (2) The primary bovine keratocytes have dendritic morphology and with limited F-action distribute in cellular cortex.

Background The incidence of posterior capsular opacification (PCO) is increasing with the growing of cataract surgery rate.Recent researches provend that disintegrin has inhibitory effect on PCO,and echistatin is one of the disintegrin prime families.Objective This study was to investigate the effects of disintegrin and echistatin on proliferation,adhestion and migration in human lens epithelial cells (LECs) line (SRA01/04).Methods Human LECs line at logarithmic growth phase was used in the study.Cells were cocultured with medium and different concentrations of echistatin (0,2.5,5.0,7.5,10.0,15.0,20.0 mg/L) for different time.The proliferative inhibitory rates of LECs were detected by MTT method 24,48 and 72 hours after cultured.Anti-adhesion effect of echistatin were analyzed by the same assay in 90 minutes.Cell scratching test was performed to evaluate the migration ability of LECs.The width of the scratch was recorded in the culture plate covered with cells under an inverted microscope.After being cultured for 24 hours and 48 hours with echistatin,cell migration distances was examined.Results Compared with the 0 mg/L echistatin group,cells proliferation was obviously inhibited.After cultured with 2.5,5.0,7.5,10.0,15.0,20.0 mg/L echistatin,the proliferation inhibitory rate was 2.6％,15.4％,21.2％,34.7％,46.1％,58.2％ at 24 hours;6.6％,21.9％,38.2％,50.0％,60.7％,76.9％ at 48 hours and 9.8％,29.0％,46.6％,63.4％,69.1％,92.4％ at 72 hours,respectively.The absorbance value (A) in the 5.0,7.5,10.0,15.0,20.0 mg/L groups were significantly lower than that in the 0 mg/L group (P＜ 0.05).With the prolongation of acting time of Ecs,the A value of the cells was gradually reduced,with statistically significant difference (P＜0.05).The adhesion inhibitory rate was 2.6％,15.0％,26.1％,35.3％,45.2％ and 54.5％ in the 2.5,5.0,7.5,10.0,15.0,20.0 mg/L group,respectively.Compared with the result in the 0 mg/L group,the A value in the 5.0,7.5,10.0,15.0,20.0 mg/L group was statistically significant (P＜0.05).After cultured for 24 hours and 48 hours,cell migration distance shortened in the 5.0,7.5,10.0,15.0,20.0 mg/L group,showing a statistically significant difference among them (P＜0.05).Cell migration distance was gradually shortened with the lapse of action time of Ecs with the significant difference (P ＜ 0.05).Conclusions echistatin has inhibitory effects on proliferation,adhestion and migration for human LECs in vitro in time-and dose-dependent manner.It is inferred that echistatin may play a role in the prevention and treatment of PCO.

Objective To investigate the standardized treatment of 33 children with thalassemia and their family financial burden registered in Bao'an district, Shenzhen city, and to provide basic information for formulating health policy for the government. Methods In 2009, preliminary investigations on 39 registered families with thalassemia children were conducted by telephone, and a household survey was made to collect treatment and economic status by questionnaire on 33 children. Results Among 33 cases of thalassemia children, 21 cases(63.7%) were severe anemia, 5 cases( 15.1%) in need of care or special care, and 25 cases(75.8%) were difficult or unable to maintain standardized treatment. The average family monthly income and expenditure was (4060 ± 2002) and (4926 ± 2991) yuan, respectively. The average monthly treatment costs were (2665 ± 1872) yuan, and the average debt amounted to (64 600 ± 53 940) yuan. Fifteen families[60.0%(15/25)] would reduce the times of blood transfusions or iron transpirations when they encountered revenue deficiency. Conclusions The heavy economic burdens on families with children thalassemia result in inadequate or interrupted treatment on sick children and affect their survival and quality of life, which should be taken more attention and social care.

Aim To investigate the role of TGF-β3 in the anti-proliferation effect of ursolic acid(UA)in co-lon cancer cells and the possible molecular mechanism underlying this effect.Methods We introduced crys-tal violet staining,flow cytometry and Western blot as-say to determine the effect of UA on proliferation and apoptosis in HCT1 1 6 cells.The levels of TGF-β3, Smad2 /3 and β-catenin in HCT1 1 6 cell were evaluated by RT-PCR and Western blot.Finally,TGF-β3 inhibi-tor and recombinant adenovirus,and luciferase reporter assay were used to analyze the possible mechanism through which TGF-β3 mediated the anti-cancer effect of UA in HCT1 1 6 cells.Results UA inhibited the proliferation and induced apoptosis apparently in HCT1 1 6 cells.UA down-regulated TGF-β3 both in mRNA and in protein level.Meanwhile,UA decreased the phosphorylation of Smad2 /3 concentration depend-ently,although no significant effect was found on the total protein level of Smad2 /3 in HCT1 1 6 cells.Over-expression of TGF-β3 attenuated the inhibitory effect of UA on the proliferation of HCT1 1 6 cells,while the TGF-β3 inhibitor potentiated this effect. UA sup-pressed the transconduction of Wnt/β-catenin signaling in HCT1 1 6 cells through decreasing the level of β-catenin.Exogenous expression of TGF-β3 increased the level of β-catenin and partly reversed the UA-in-duced decrease of β-catenin.However,TGF-β3 inhib-itor potentiated the inhibitory effect of UA on β-catenin in HCT1 1 6 cells.Conclusion The anti-proliferation activity of UA in colon cancer may be partly mediated through down-regulating TGF-β3 to suppress Wnt/β-catenin signaling at least.