Objective To master the technique of mouse embryonic stem (ES) cells differentiate into endothelial cells,which would be a new therapeutic approach for cardiovascular disease.Methods Expression of selfrenewal marker genes in E 14 cells was assessed.Expression of vascular endothelial growth factor receptor 2 (Flk 1) in monolayer differentiation on day 4 and vascular endothelial cadherin (VE-cadherin) on day 8 were detected.On day 8,differentiation cells were also observed under phase contrast microscopy (PCM) and transmission electron microscope (TEM).ES cells and endothelial-specific molecular markers were assessed by RT-PCR at different time-points.Results As self-renewal marker genes were expressed in E14 cells,E14 cells was identified to maintain their selfrenewal pluripotency.The marker gene of letarl,Flk1 was expressed on differentiation day 4.On differentiation day 8 the marker gene VE-cadherin was expressed and as observed under PCM endothelial cells with spindle shape and TEM with Weibel-Palade body,thus were the major populations generated after VEGF induction,and E14 cells were confirmed differentiated into mature endothelial cells.The expressions of genes octamer binding transcription factor 4 (Oct4),Flk1 and VE-cadherin were detected on differentiation day 2,4,6,8 and 10.Conclusions As VE-cadherin gene was expressed in monolayer on differentiation day 8,E14 cells were confirmed differentiated into endothelial cells,which would be a new therapeutic approach for cardiovascular disease.

Objective To investigate pathogenic factors,clinical characteristics,pathological changes in 6 patients with localized or disseminated intestinal neuronal dysplasia(IND).Methods The clinical retrospective study was performed in 6 cases with localized and disseminated IND.Results In 6 patients,constipation and partial intestinal obstruction were the major symptoms.Preoperatively,dia-gnoses were revaluated as intestinal stenosis in 2 cases,long sigmoid in 2 cases and long-segment aganglionosis and total gastrointestinal IND in the rest 2 cases,respectively.Exploratory laparotomy was performed on all these patients.Pathological study suggested that 4 of them were localized IND,on whom enterectomy and intestinal anastomosis were performed successfully.However,2 cases of disseminated IND died postoperatively.Conclusions Disseminated IND may be caused by congenital factors,while localized IND by the acquired factors which including intestinal obstruction,local ischemia or inflammation.The surgical interventions are effective for localized IND,and intestine transplantation might be prospective treatment for disseminated IND.

Objective To investigate the safety and efficacy of structured triglycerides in parenteral nutrition in elderly patients with acute biliary tract infection.Methods 62 elderly patients with acute biliary tract infection under conservative therapy were randomly divided into structured triglycerides (ST) group and medium-chain triglycerides plus long-chain triglycerides (MCT/LCT) group.Patients underwent parenteral nutrition for 5 days.Levels of hepatic enzymes,blood triglycerides,nitrogen balance,fasting blood insulin and glucose were compared between the two groups after 5 days of parenteral nutrition.Results There were no significant differences in levels of hepatic enzymes,blood triglycerides and fasting blood glucose between ST group and MCT/LCT group.Plasma total bilirubin (T-Bill) levels were increased in the two groups,and T-Bill level dropped faster in the ST group than in the MCT/LCT group after 7 days of parenteral nutrition(P＜ 0.05).The difference in total cholesterol level dropping was significant between ST group and MCT/ LCT group (P＜0.05).The cumulative nitrogen balance for 5 days had significant difference between ST and MCT/LCT groups [(4.7-±-2.2) g vs.(2.8±0.7) g,P＜0.05].STG group versus MCT/LCT group had less fluctuation of fasting blood insulin level after 1,4 and 7 days of parenteral nutrition.Conclusions In the acute phase of biliary tract infection in elderly patients,ST-based parenteral nutrition can provide energy and help maintain nitrogen balance,with a little fluctuation of fasting blood insulin level.Short-term nutritional support may have better tolerance in the liver and has little effect on the change of hepatic enzymes.

Objective To observe the effect of fenofibrate intervention on high-fructose-feeding-induced liver steatosis in rats and explore the possible mechanism. Methods Male Wistar rats were randomly divided into control group ,high fructose group and fenofibrate group[fenofibrate intervention started after 8 weeks of high fructose feeding ,30 mg/(kg · d)]. Rats were sacrificed after 12-week of high fructose feeding. Serum alanine aminotransferase(ALT),aspartate aminotransferase(AST),total cholesterol(TC),free triglyceride(TG)and liver TG content were determined;protein levels of fatty acid synthase(FAS),endoplasmic reticulum stress mark-er Bip and autophagy markers such as Atg7,Beclin1,LC3 and the related pathway mTOR in liver tissues were de-tected. Results Compared with those in control group and fenofibrate group,serum AST,serum total cholesterol, blood free TG and hepatic TG were significantly increased in high-fructose group(P < 0.01). The protein expres-sion of Fas,Bip and mTOR were significantly increased in high-fructose group compared with those in control group and fenofibrate group;the protein expression of Atg7,beclin1 and LC3 were significantly decreased in high-fructose group compared with those in control group and fenofibrate group. Conclusions Long-term high-fructose-feeding induces fatty liver and liver cell injury ,and may affect ERS and autophagy. High-fructose-feeding-in-duced fatty liver may be improved by fenofibrate and its underlying mechanism might be associated with Fas,ERS and autophagy in liver.

Objective To investigate the distribution of measured value and the expected value of spirometry parameters in healthy children aged 5-14 years old in Beijing,and to provide evidence for clinical interpretation precisely.Methods Four hundred healthy children (198 boys and 202 girls) aged 5 to 14 years from the kindergartens and the primary schools in Beijing were selected as subjects.All of the parameters were measured by using Jaeger Masterscreen spirometry machine(Germany).Pulmonary function parameters including forced vital capacity(FVC),forced expiatory volume in 1 second (FEV1),FEV1/FVC,peak expiratory flow(PEF),forced expiratory flow at 50% of vital capacity(FEF50),forced expiratory flow at 75% of vital capacity(FEF75),and the maximal midexpiratory flow(MMEF) were collected for analysis.The expected values of the 7 parameters above were calculated according to Zapletal prediction equations(4-18 years old).The measured value vs.the expected value was calculated.Results Children aged 5 to 12 years had similar height and weight between boys and girls,and the differences were not significant(all P>0.05);while boys were taller and heavier than girls in 13-<14 years old group and 14 years old group,and the diffe-rences were significant(t=3.35,2.05,5.99,2.67;all P<0.05).The mean measured value vs.the expected value of PEF,FVC,FEV1,FEV1/FVC of children in different gender and age groups ranged between 105.0%-114.7%,95.4%-106.8%,104.6%-115.4% and 105.8%-112.6%,respectively.The mean measured value vs.the expected value of small airway function parameters FEF50,FEF75,MMEF was as follows respectively:79.7%-103.7%,70.2%-110.5%,and 81.6%-114.7%.The low limit of normal(LLN) measured value vs.the expected value of the 7 pulmonary function parameters was as follows:PEF 85.0%,FVC 84.7%,FEV1 92.4%,FEV1/FVC 98.4%,FEF50 67.7%,FEF75 60.1%,and MMEF 71.0%.Conclusions The pulmonary function parameters of the healthy children in Beijing are higher than those of the Zapletal prediction.The measured value vs.the expected value of FEV1 92.4%,FEV1/FVC 98.4%,PEF 85.0%,FVC 84.7%,FEF50 67.7%,FEF75 60.1%,and MMEF 71.0% can be used as LLN in 5-14 years old children.

BACKGROUND: Literatures report that double-bundle reconstruction is a better choice for posterior cruciate ligament (PCL),but Tibial-inlay technique exhibits no significant difference.OBJECTIVE: To evaluate the effect of Tibial-inlay technique used in the PCL reconstruction with allograft tendons.METHODS: Thirty-one patients with PCL rupture were verified by arthroscopy in the Department of Orthopaedics, at the Affiliated Hospital of Chengde Medical College between February 2006 and May 2008, including 14 knees caused by traffic accident injury,9 knees by crashing, and 8 knees by athletic injury. All the damaged PCLs were reconstructed with allograft tendons by Tibial-inlay technique under arthroscopy. During surgery, the remnants of the original PCL were retained as much as possible,PCL femoral tunnel was prepared. By using of allogeneic patella tendon with bone block at both ends or achilles tendon allograft with bone block at one side, the lateral bone block was chipped into trapezoidal block at a width of 1.0-1.5 cm and a length of 2 cm, that is, in addition to relying on friction force to fix between bone groove and bone graft, it can also generate compressive stress and increase its stability. The PCL tibial attached point was stripped, the trapezoid bone groove was prepared according to the size of bone block at the insertion site, and allogeneic bone block was embedded into bone groove, then the other end was introduced to femoral tunnel using pulling wire through the joint, after tensed at anterior drawer site, the extrusion screw was twisted forward to fix the tendon or bone. Before operation and during follow-ups, the posterior drawer test knee instability, joint activity, Lysholm knee function scores were recorded.RESULTS AND CONCLUSION: No severe complications, such as vascular nerve injury, rejection or infection, occurred in early stage after operations in 31 patients. All of them were followed up for 12-24 months. Posterior drawer test of all the subjects were above 2+ before operation, of which, 3+ and 4+ were 84%; the posterior drawer test re,suits were as the following after operation:4 cases of 0+ (normal), 17 cases of 1 +, 9 cases of 2+ and 1 case of 3+, which shows the improvement of retroposition (P ＜ 0.05);Lysholm score was remarkably improved at follow-up compared with preoperation (P ＜ 0.05), and there were significant improvements in the joint range of motion (P ＜ 0.05). The reconstruction of PCL by Tibial-inlay technique with allograft tendons has advantages of minimal trauma in surgery, reliable fixation and satisfactory outcome.

To demonstrate the current strategies for treating cartilage defects of knees and the related research. Published papers about cartilage defects were searched and reviewed. The current strategies for the treatment were summarized. Based on the research of our study and others, the conclusion how to treat cartilage defects was made. The current ways for treating cartilage defects include micro-fractures, chondrocytes transplantation, mosaicplasty and tissue engineering; Research on functional magnetic resonance imaging in the early diagnosis of cartilage defects, cartilage degeneration is gradually increasing. There is still no effective treatment of cartilage defects and tissue engineering has brought new hopes for the treatment of cartilage defects , functional magnetic resonance imaging has some significance in early diagnosis of cartilage defects, cartilage degeneration.
Animals ; Cartilage Diseases ; surgery ; therapy ; Cartilage, Articular ; surgery ; Humans ; Knee ; surgery ; Tissue Engineering ; Transplantation, Autologous

Objective To discuss the relationships between serum uric acid levels and clinical stages,cognitive dysfunction in parkinson' s disease (PD).Methods Sixty patients of PD were enrolled,and their history,detailed physical examination UPDRS Ⅲ scale and mini-mental state score (MMSE) scale were performed to determine the Hoehn-Yahr stage of patients.At the same time 52 cases of healthy person were collected from the Health Examination Centers whose age,sex,and education matched with PD group as control group.The levels of serum uric acid in the two groups were examined and compared.Results The serum uric acid levels in PD group were significantly lower than those in normal control group((262.4 ±85.2) μmol/L vs (357.6 ±73.3) μmol/L) (t =1.013,P =0.005) ;There was no level correlation between different Hoehn-Yahr stage period of PD group (r =0.143,P ＞ 0.05) ; There was no significant difference on serum uric acid levels between subgroup of PD patients (F =1.014,P =0.06) ;The M MSE score between PD group and normal control group had no significant difference (26.12 ± 2.87 vs 28.32 ± 2.17) (t =1.970,P =0.073).Conclusion Low level of the serum uric is likely to be a risk factor for PD,but cannot be used to assess the severity of disease and MMSE scale used to determine the cognitive impairment in PD is not sensitive enough.

Objective To investigate the condition and the related influence factors of small airway function in controlled bronchial asthmatic children.Methods Children diagnosed with bronchial asthma controlled,aged 5-14 years old without gender limitation were consecutively enrolled.Survey questionnaire including asthma symptoms in the past one month and medication usage were conducted.Physical condition and lung function were examined.Results A total of 255 patients were included in this study,and 224 patients had normal small airway function in which the level of forced expiratory flow at 50％ of forced vital capacity(FVC) exhaled (FEF50),forced expiratory flow at 75％ of FVC exhaled(FEF75),maximal mid-expiratory flow(MMEF) was 87.9％ (77.0％,97.2％),73.8％ (65.6％,93.5％),and 84.9％ (76.4％,97.5％) of the predicted value,respectively.Thirty-one patients had small airway dysfunction,accounting for 12.2％ (31/255 cases),and the level of FEF50,FEF75,MMEF were 62.8％ (59.9％,65.5％),51.9％ (46.6％,55.5％),and 62.7％ (57.2％,64.4％) of the predicted value,in which FEF75 had shown more decrease than others.In small airway dysfunction group,10 patients had conducted bronchodilation testing,and improvement rate of FEF50,FEF75 and MMEF were 41.1％,47.5％ and 41.3％,and all 10 cases (100.0％) returned to normal level.No significant difference was found in gender,body mass index,atopy,disease duration,drug usage between 2 groups (all P ＞ 0.05).Conclusion In 5-14 controlled asthmatic children,12.2％ have small airway dysfunction with a certain degree of reversibility.Gender,body mass index,atopy,disease duration,drug usage are not influencing factors.

OBJECTlVE To study the reIationship between microRNA(miRNA)-122 and Iiver injury in-duced by anti-tubercuIosis drugs,and to discover the new biomarkers for earIy diagnosis of this type of Iiver injury. METHODS mice were given 2 mL isoniazid oraIIy at 90 mg·kg-1 . BIood and Iiver tissue sampIes were coIIected at 1,3,5,7,14,21 and 28 d after administration of isoniazid. Serum gIutamic-pyruvic transaminase( GPT)and gIutamic-oxaIoacetic transaminase( GOT)IeveIs were determined using an automatic biochemicaI anaIyzer. Cu/ Zn-superoxide dismutase( Cu/ Zn-SOD ) activity and maIondiaIdehyde( mDA)content were detected by biochemicaI method. ReaI-time qPCR was used to measure the expression of miRNA-122. RESULTS GPT and GOT IeveIs were significantIy higher at 14 and 21 d(P﹤0.05)than in the controI. Cu/ Zn-SOD began to decIine whiIe mDA began to increase after 5 d(P﹤0.05). miRNA-122,which progressiveIy decreased after administration,was reduced to the mini-mum 0.58 ±0.02 at 14 d. There were good correIations between miRNA-122 and GPT,Cu/ Zn-SOD, mDA(the correIation coefficients were -0.370,0.268,and -0.298,respectiveIy),but no correIation with GOT was observed. CONCLUSlON The tissue miRNA-122 profiIe can be used as a sensitive marker for anti-tubercuIosis drug-induced Iiver injury,which couId contribute to the earIy diagnosis of Iiver injury.