Objective To evaluate the platelet function in the patients with recurrent spontaneous abortion (RSA) ,to explore the correlations between Hcy and folic acid(FA)with mean platelet volume(MPV) and platelet distribution width(PDW) ,and to evalu-ate the application value of above indicators in predicting RSA .Methods Twenty-eight cases of RSA ,40 cases of healthy pregnancy and 50 healthy child-bearing women(control group) in our hospital were selected .The detection indicators included peripheral blood Hcy ,FA ,PLT ,MPV and PDW .Results The PLT and FA levels in the patients group were significantly lower compared with the normal pregnancy group and control group(P<0 .01) ,while the Hcy ,PDW and MPV were increased(P<0 .05);the various indica-tors had no statistically significant difference between the normal pregnancy group and control group (P>0 .05) .Blood Hcy was negatively correlated with PLT and FA ;Hcy and FA were negatively correlated in the normal pregnancy group ,other indicators had no obvious correlation(P>0 .05) .Conclusion The Hcy metabolism abnormality ,platelet parameter changes and their interactions may be related to RSA .The combined detection of Hcy ,FA and platelet parameters has a certain clinical significance to early diag-nosis and treatment of RSA .

Objective The stability and other characteristics of the active recombinant human anionic trypsin(hT 2) with site-mutation R 122 L(mhT 2) were investigated. Methods An active human anionic trypsin and its R 122 L mutate were produced with E.coli BL 21(DE 3) and purified with ion-exchange chromatography. The properties of mutant were studied and compared with the wild type. Results The optimal pH for mhT 2 was 7~11. mhT 2 was active over a broad temperature range (4℃~80℃) and owned a little better thermal stability than the wild type. The inhibition of typical metal chelating agent(EDTA), Fe 3+, denaturant, reducer(β-ME) on activity of mhT 2 was the same as the wild type. Michaelis constant Km of mhT 2 was 0.010 mmol/L with BAEE as a substrate, a little lower than wild type. Conclusion Compared with the wild type, the R 122 L site mutate significantly enhanced tolerance to acidic pH、denaturants、reductions and autolysis.

BACKGROUND:At present, there are no studies of comparing the effect of silk fibroins from different sources in repair of osteochondral defects. OBJECTIVE:To compare the effect of mulberry silk- and tussah-derived silk fibroin scaffold materials in repair of osteochondral defect. METHODS:Totaly 20 New Zealand white rabbits were obtained to prepare osteochondral defect models on the unilateral knee joint and randomly divided into five groups: control group, experimental group 1, experimental group 2, experimental group 3 and experimental group 4. Rabbits in the control group were not implanted any materials. In the experimental group 1, 3 layers of mulberry silk protein scaffolds stuck together to fil in defects. In the experimental group 2, one mulberry silk protein scaffold coated with transforming growth factor-β3 was stuck with two mulberry silk protein scaffolds coated with bone morphogenetic protein-2 to fil in defects. In the experimental group 3, three layers of tussah protein scaffolds stuck together to fil in defects. In the experimental group 4, one tussah protein scaffold coated with transforming growth factor-β3 stuck together with two tussah protein scaffolds coated with bone morphogenetic protein-2 to fil in defects. At 8 weeks post surgery, articular cartilage repair area was observed histopathologicaly. Type I and II colagen expressions were determined. RESULTS AND CONCLUSION:The colagen fibers in experimental group 1 were widely distributed in the ful-thickness defect area. The colagen fibers in the experimental group 2 were paralely distributed on the surface of repair area, verticaly distributed from the middle and bottom to the top direction. Colagen was observed on the surface of repair area in the experimental group 3. The cartilage-like cels presented clumped distribution on the surface and at the bottom of scaffold. The type I colagen expression in the repair area was strongly positive in these four experimental groups. The type II colagen expression in the repair area of experimental 1 and experimental 2 groups was weak. The type II colagen expression in the repair area of experimental 3 and experimental 4 groups was strongly positive. These results demonstrate that these two kinds of silk fibroins can both repair osteochondral defects, in which mulberry silk proteins tend to form bone tissue, and tussah silk proteins tend to form cartilage tissue.

Objective To observe the efficacy and toxicity of 5 -Fluorouracil plus cisplatin (FP)and docetaxel plus cisplatin(DP)in the treatment of patients with metastatic esophageal cancer.Methods A total num-ber of 68 cases of metastatic esophageal cancer confirmed by histopathology and (or)cytological examination was ana-lyzed retrospectively.There were 33 cases in FP group(5 -Fu 500mg/m2 ,d1 -5;DDP 20mg/m2 ,d1 -5,28 days a cycle),and 35 cases were in DP group(DTX 75mg/m2 ,d1,DDP 20mg/m2 ,d1 -3,21 days a cycle).The evaluation of efficacy and toxicity were performed on all patients per 2 cycles.Statistics and analysis of the difference between the two groups was performed.Results For FP group,CR was in 1 case,PR ere in 12 cases,SD ere in 16 cases and PD ere in 4 cases,respectively.The corresponding value was 3,18,11 and 3 for DP group.The objective response rate (ORR)for DP group(68.57%)was significantly higher than FP group(36.39%)(χ2 =5.83,P =0.02),and the disease control rate (DCR)were similar with no significance between the two groups(P =0.71).The Ⅲ -Ⅳ degree myelotoxicity for DP group was higher than for FP group with statistical difference (χ2 =7.07,P =0.01),while the incidence of nausea,vomiting,diarrhea and liver toxicity was similar (P >0.05).Conclusion DP regimen was more effective than FP regimen for the metastatic esophageal cancer,and the patients can tolerate with the slightly higher incidence of myelotoxicity,it was worthy of further study and clinical application.

Aim To observe the effects of theaflavin on intracellular calcium concentration(i) level in rat ventricular myocytes and discuss the possible mechanisms.Methods The effects of theaflavin oni were investigated in rat ventricular myocytes.i was detected by laser confocal microscopy and represented by relative fluorescent intensity(FI-FI0)/FI0,%;FI0:control;FI:administration of drugs).Results ① Theaflavin(10,20,40 ?mol?L-1) had no effect on the i of ventricular myocytes in normal Tyrode′s solution.However,it reduced the i of ventricular myocytes in simulated ischemia solution in a concentration-dependent manner.② Pretreatment with Bay k8644(0.5 ?mol?L-1) mostly abolished the effects of theaflavin(20 ?mol?L-1) in simulated ischemia solution.③ Theaflavin(20 ?mol?L-1) markedly inhibited the low concentration of ryanodine-induced i increase in Ca2+-free Tyrode′s solution.④ When the propagating waves of elevated i(Ca2+ waves) were produced by increasing extracellular Ca2+ concentration from 1 mmol?L-1to 10 mmol?L-1,theaflavin(20 ?mol?L-1) could block the propagating waves of elevated i(Ca2+ waves),reduce the frequency and duration of propagating waves,and reduce i as well.Conclusion Theaflavin may reduce the i in isolated rat ventricular myocytes via inhibiting Ca2+ influx by voltage-dependent Ca2+ channel and alleviating Ca2+ release from sarcoplasmic reticulum(SR).

OBJECTIVEThe glucocorticoid dexamethasone (DEX) can induce palatal cleft; however, the mechanism involved remains unclear. E-cadherin is an important cell adhesion molecule, and it can significantly affect cell fate and embryonic development. Recent studies have indicated that E-cadherin expression in palatal epithelial cells is suppressed in normal palate fusion. This study aimed to determine whether the change in E-cadherin expression is related to the incidence of cleft palate in DEX-induced mice.

METHODSMice were divided into the experimental group and the control group. Pregnant mice were injected with DEX on E10.0-E12.0, whereas mice in the control group were injected with normal saline. Hematoxylin and eosin (HE) staining, immunohistochemistry, and real-time quantitative polymerase chain reaction were employed to evaluate the effect of DEX on fetal mouse palatal processes, particularly the changes in E-cadherin and β-catenin expression levels in the phases of the experimental and control groups.

RESULTSData indicated that the incidence of cleft palate in the DEX group was 43.59% (17/39), whereas that in the control group was only 3.03% (1/33). The results of HE staining showed that the obviously shortened palatal processes could not contact and fuse with one another in the DEX-treated mice model compared with those in the control group. The ectopic expression of E-cadherin in embryonic palatal mesenchymal cells was also analyzed. The expression levels of E-cadherin and β-catenin in the experimental group were higher than those in the control group.

CONCLUSIONThese findings indicated that DEX could induce E-cadherin gene upregulation and ectopic expression, as well as high β-catenin expression, thereby inhibiting the growth of mesenchyme cells and cleft palate.

Animals ; Cadherins ; genetics ; metabolism ; Cleft Palate ; chemically induced ; embryology ; Dexamethasone ; adverse effects ; Disease Models, Animal ; Epithelial Cells ; Female ; Glucocorticoids ; Immunohistochemistry ; Mice ; Pregnancy ; beta Catenin ; metabolism

BACKGROUND: Insulin resistance (IR) is associated with type 2 diabetes mellitus and its macrovascular complications. Euglycemic agent (rosiglitazone and metformin) can ameliorate IR, but its influences on other cardiovascular risk factors, including blood lipid, blood pressure and body mass need to be further studied.OBJECTIVE: To observe the effects of rosiglitazone and metformin on the blood glucose, blood lipid, blood pressure and body mass of patients with type 2 diabetes mellitus complicated by blood lipid disturbance.DESIGN: A clinical observation.SETTING: Department of Endocrinology, Nanjing First Hospital, Nanjing Medical University.PARTICIPANTS: Forty-two patients with type 2 diabetes mellitus complicated by dyslipideamia, who received treatment in the Department of Endocrinology, Nanjing First Hospital, Nanjing Medical University between June and September 2006 were recruited in this study. The involved patients, 27 male and 15 female, averaged (58±2)years ranging from 30 to 70 years old. Inclusive criteria:①Type 2 diabetes mellitus all met the diagnostic criteria of American Diabetes Association (2006).② Diagnostic criteria of blood lipid disturbance : total cholesterol(TC)＞ 4.68 mmol/L, triglyceride (TG) ＞ 1.7 mmol/L, low density lipoprotein cholesterol (LDL-C) ＞ 2.68 mmol/L, high density lipoprotein cholesterol (HDL-C) ＜ 0.9 mmol/L (female), ＜ 1.0 mmol/L (male). Informed consents were obtained from all the patients.METHODS：①Drug treatment：Dudng the 12-week treatment,all the patients received 0．75 g metformin(Lot No．051110 and 051214,0.25 g，lablel,Jiangsu Hengrui Medicine Co.，Ltd)and 4 mg rosiglitazone (Lot No．0511101,2 mg／tablet，Jiangsu Hengrui Medicine Co.，Ltd) daily．②Physical and laboratory measurements：Body mass index，blood pressure were taken，and blood lipid profile(TC,TG,C-LDL and C-HDL)，fasting and postprandial blood glucose were measured by colodmetdc method before,8 and 12 weeks after treatment．MAIN OUTCOME MEASURES：①Fasting and postprandial blood glucose level change．②Blood lipid，blood pressure and body mass index change．RESULTS：Forty-two patients with type 2 diabetes mellitus complicated by dyslipideamia were recruited，and three dropped out the experiment，one patient suffered from severe watery stool,one patient from severe abdominal distention,one patient from urinary tract infection, and the other thirty-nine patients completed the observation in the final analysis．Compared with pretraatment data,fasting and postprandial blood glucose levels 12 weeks after treatment were decreased(P＜0．01),TC increased(P＜0．05)，LDL-C decreased(P＜0．01)，HDL-C increased(P＜0．01),blood pressure did not change(P＞0．05) and body mass index increased(P＜0．01)．CONCLUSION：Combinatin of rosiglitazone and metformin significantly decreases blood glucose，decreases LDL-C,increases TC，HDL-C,body mass index,and has no obvious effects on TG and blood pressure．It is safe and bearable for type 2 diabetes mellitus patients with dyslipideamia．

Objective:To observe the clinical efficacy and safety of Bianyan oral liquids in the treatment of acute pharyngitis. Methods:The clinical data of 200 patients with acute pharyngitis in the outpatient department of traditional Chinese medicine in our hospital from September 2010 to June 2012 were analyzed. They were divided into the control group with 100 cases and the observation group with another 100 cases. The control group was given Jinlianhua granules, while the observation group was received Bianyan oral liquids. After 5 days, the symptom score, effective rate, changes in accompanied symptoms and safety index were observed and com-pared. Results:The symptom score and effective rate of the observation group were better than those of the control group (P<0. 05), and the improvement in defecation was better than that of the control group (P<0. 05) as well. There were two cases in the observation group suffered nausea and stomachache, and one case in the control group suffered diarrhea while no untoward effect emerged. Conclu-sion:Bianyan oral liquids with the effects of heat-clearing and detoxicating, nourishing yin and fluid and treatment both manifestation and root cause show significant effect in the treatment of acute pharyngitis.

Objective To observe effects of the antisense genes of Toll-like receptor 4 (TLR4 ) and myeloid differentiation protein-2 (MD2) on the activation of NF-?B in alveolar typeⅡepithelial cells (ATⅡcells) induced by lipopolysaccharide (LPS). Methods Cultured ATⅡcells were randomized to normal cells (control) group,LPS group,LPS+empty vector group,LPS+TLR4 antisense gene group,LPS+MD2 antisense gene group,LPS+TLR4-MD2 antisense gene group. The expressions of TLR4 and MD2 mRNA and protein in ATⅡcells were detected by Northern blotting and Western blotting respectively. The activity of NF-?B in ATⅡcells was measured with electrophoretic mobility shift assay (EMSA). The TNF-? and IL-6 concentrations in the supernatant of cultured ATⅡcells were tested with ELISA.Results Compared with control groups,the expressions of TLR4 and MD2 mRNA and protein,the NF-?? activity,the levels of TNF-? and IL-6 were increased remarkably in LPS group and LPS+empty vector group (P

Recent evidence suggests Toll like receptor 4 (TLR4) and its accessory protein MD2 mediate endotoxin/lipopolysaccharide (LPS) signaling. LPS binds to LPS binding protein (LBP) in plasma and is delivered to CD14. Next, LPS is transferred to TLR4 MD2 complex. LPS activates several intracellular signaling pathways that include the NF ?B pathway and three mitogen activated peotein kinase (MAPK) pathway: extracellular signal regulated kinase (ERK), c Jun N terminal kinase (JNK) and p38. These signaling pathways in turn activate NF ?B and AP 1 (c fos/c jun), which coordinate the induction of many genes encoding inflammatory mediators. Targeting the signaling molecule in the pathway will develop new remedies for treatment of inflammatory disorder.