ObjectiveTo investigate the effects of carboxymethyl-chitosan(CMCS) on chondrocyte apoptosis induced by sodium nitroprusside (SNP),and the effects of CD44 in the process.MethodsA small interference RNA (siRNA) targeting to CD44 mRNA (siRNA-1,siRNA-2,siRNA-3) was constructed.The siRNA was transfected into chondrocytes in vitro with LipofectamineTM 2000.The efficacy of transfection was detected by transfecting fluorescence siRNA into cells.The mRNA expression of CD44 in vitro was detected by RT-PCR.The protein level of CD44 was detected by Western blotting.The apoptosis rate of the transfected and non-transfected cells induced by SNP was detected by flow cytometry.Statistical analysis was conducted with one-way ANOVA and SNK-q test.ResultsThe efficacy of transfection was about 60％.As compared with the control group,the mRNA expression was specifically inhibited after transfecting CD44 siRNA-1 for 24,48 and 72 h(0.198±0.007 vs 0.429±0.053 at 24 h,0.211±0.016 vs 0.501±0.037 at 48 h,0.153±0.005 vs 0.341±0.009 at 72h,q=5.93,7.01,11.23,P＜0.01 ),and the protein level of cells was inhibited after transfecting CD44 siRNA-1 for 24 h compared with the control group (0.231±0.064 vs 0.675±0.113,q=13.09,P＜0.01 ).The FCM results showed that 3 mmol/L SNP could induce chondrocytes apoptosis(70±6)％,and 50,100,200 μg/ml C MCS could affect the inhibitory effect of SNP-induced apoptosis of chondrocyte [ (51 ±7)％,(30±4)％,(15±4)％,q=5.08,6.97,9.73,P＜0.01 ],but it had milder inhibitory effect on CD44-siRNA-1 transfected chondrocytes when compared with those of the non-transfected chondrocytes [ (34±6)％ vs(15±4)％,q=6.95,P＜0.01 ].ConclusionThe data of this study has suggest that siRNA-1 against CD44 gene can significantly inhibit the expression of CD44 in chondrocyte of rats in vitro after transfection.The CD44 may play an important role in chondrocyte apoptosis induced by SNP and protected by CMCS.

Objective To study the level changes and correlation of homocysteine (Hcy) ,high-sensitivity C-reactive protein(hs-CRP) ,cystatin C(Cys-C) and plasma fibrinogen(Fib) in the patients with acute cerebral infarction (ACI) .Methods The fasting blood samples were collected from 178 cases of ACI(ACI group) and 93 healthy individuals blood samples (control group) .The lev-els of serum Hcy ,hs-CRP and Cys-C were detected by the BECKMAN AU-680 fully automatic biochemical analyzer and plasma Fib was determined by the RAC-100 fully automatic coagulometer .Results The levels of serum Hcy ,hs-CRP ,Cys-C and plasma Fib in the ACI group were significantly increased compared with the control group (P<0 .05) .There was significantly positive correlation between Hcy with hs-CRP and Cys-C in the ACI group(r=0 .326 ,0 .361 ,P<0 .05) ,but there was no significant correlation be-tween Hcy and Fib ;there was significantly positive correlation between hs-CRP with Cys-C and Fib(r=0 .365 ,0 .421 ,P<0 .05);the same significant positive correlation also existed between Cys-C and Fib (r=0 .447 ,P<0 .05) .The positive rate of the joint de-tection of Hcy ,hs-CRP ,Cys-C and Fib was 93 .8 % ,which was obviously higher than that of the single indicator detection (P<0 .05) .Conclusion Cys-C ,hs-CRP ,Fib and Hcy participate in the occurrence and development process of ACI ,their joint detection has the important clinical significance for the prevention ,early diagnosis and treatment of ACI .

Hepatocellular carcinoma(HCC)is one of the most common malignancies worldwide.Due to the difficulty of diagnosis in the early stage of HCC, most HCCs are diagnosed in intermediate-advanced stage.Moreover, the high invasion, metastasis and recurrence rate of HCC result in the high mortality of HCC.MicroRNAs(miRNAs) are a class of highly conserved, endogenous, small, non-coding ,single stranded RNA with the length of 22 nucleotides.There are plentiful of miRNAs in liver.MiRNAs not only can regulate the growth and development of liver, but also are closely related to the formation of HCC.In the process of HCC formation, miRNAs could function as oncogenes or tumor suppressor genes to regulate multiple biological processes related to HCC, including cell differentiation,proliferation,tumorigenesis,angiogenesis,invasion,and metastasis.With the intensive study of molecular mechanisms of miRNAs in the process of HCC formation, increasingly studies have revealed that miRNAs could become sensitive biomarkers and effective therapeutic targets for HCC.

Objective To study the diagnosis,surgical treatment and prognosis of patients with primary duodenal carcinoma.Methods The clinical data of 56 patients with primary duodenal carcinoma treated between 2008 to 2015 were retrospectively analyzed.Results The number of patients with tumors located in the first,second,third and fourth parts of duodenum were 3,44,6,and 3 patients respectively.Tumors which were within the papillary region accounted for 71.4％ (40 patients).Twenty-two patients (22/56,39.3％) had well differentiated adenocarcinoma,16 patients (16/56,28.6％) had moderately differentiated adenocarcinoma and 6 patients (7/56,10.7％) had undifferentiated carcinoma.The clinical manifestations were not specific,which included abdominal pain,abdominal distention,jaundice,bowel obstruction or bleeding.The correct rates of diagnosis made by endoscopy,duodenography,ultrasound and CT were 84.0％,81.3％,30.4％ and 48.2％ respectively.Forty patients underwent pancreaticoduodenectomy,5 segmental duodenectomy,9 bypass operation,and 3 subtotal gastrectomy and duodenal bulb tumor resection.The 1-,3-,and 5-year survival rates of all the patients were 82.6％,56.7％ and 30.1％ respectively.The-1,3-,and 5-year survival rates of the patients who underwent pancreaticoduodenectomy and segmental duodenectomy were 100％,68.8％,42.2％ and 100％,61.8％,0 respectively.All the patients who underwent palliative resection died 6 to 24 months after surgery.Univariate analysis revealed the operation types.,depth of tumor invasion,lymphatic invasion,and tumor differentiation correlated with prognosis.Multivariate analysis showed only the operative types,depth of tumor invasion and lymphatic invasion to be independent prognostic factors.Conclusions Tumors located in the papillary region accounted for the majority of primary malignant tumors of the duodenum and they were mainly adenocarcinomas.Duodenography and endoscopy were the major methods used in the diagnosis of primary duodenal carcinoma.Pancreaticoduodeneetomy was the best therapy for primary duodenal carcinoma.

Objective To observe the impact of diabetes and stress hyperglycemia on thrombolytic effect and short-term prognosis in patients with acute cerebral infarction. Methods A total of 127 patients with acute cerebral infarction (≤4. 5 h) who received thrombolytic therapy with alteplase at General Hospital of Beijing Military Command from January 2012 to August 2013 were enrolled retrospectively. They were divided into three groups:Diabetes group (n=35),stress hyperglycemia group (n=49),and normal glucose group (n=43) according to whether they had a history of diabetes,random glucose on admission, and oral glucose tolerance test at day 7. At 24 h after thrombolysis,the National Institute of Health Stroke Scale (NIHSS) scores,recanalization rate,and the modified Rankin Scale (mRS) scores at day 90 were compared between the 2 groups. Results Before thrombolysis,the NIHSS scores of the diabetic group, stress hyperglycemia group,and normal glucose group were 14. 2 ± 5. 1,12. 8 ± 5. 6,and 13. 0 ± 4. 6,respectively (P>0.05);at 24 h after thrombolysis,they were 14.7 ±6.0,11.9 ±4.9,and 8.0 ±2.9,respectively (P<0.05);compared with before thrombolysis,the NIHSS scores of the diabetes group and the stress hyperglycemia group had no significant change (P>0. 05);the NIHSS score of the normal glucose group was lower than before thrombolysis. There was significant difference (P <0. 05). After thrombolysis,the patients with good recanalization were 54. 3% (n=19),57. 1% (n=28),and 67. 4% (n=29),respectively in the three groups;the hemorrhagic conversion rate was 14. 3% (n=5),6. 1% (n=3),and 2. 3% (n=1),respectively. There were no significant differences. At day 90 after thrombolysis,the mRS scores in the 3 groups showed that the good prognosis rate of the normal glucose group was 72. 1% (n=31);it was significantly higher than 51. 0% (n=25) of the stress hyperglycemia group and 29. 6% (n=10) of the diabetes group. There were significant differences (P<0. 05,P<0. 01). There was also significant difference between the stress hyperglycemia group and the diabetes group. Conclusion Diabetes and stress hyperglycemia have varying degrees of adverse effects on the efficacy and prognosis of the thrombolytic therapy for acute cerebral infarction.

Objective To investigate the modulating effects and explore their mechanism of 9-nitrocamptothecin and its liposomes to induce apoptosis and inhibit cell cycle in HepG2 and L02 cell lines. Methods Cells were incubated with 9-nitrocamptothecin(9NC) or with 9-nitrocamptothecin liposomes for 24 h, 48 h and 72 h, then, the cell viability was measured via MTT assay; cell cycle and apoptosis was evaluated by flow cytometry after stained by PI and Annexin V-PE/7AAD. Additional, Western Blot was used to evaluate the expression of cell cycle and apoptosis related protein. Results Both cells viability were apparently inhibited by the 9-nitrocamptothecin and 9-nitrocamptothecin liposomes, the inhibitory effect showed a time-dependent and dose-dependent manner. Both S and G2/M phases arrest were observed after incubated with drugs. HepG2 cell was completely arrested in S phase when 9NC concentration over than 0. 1 μmol/L after incubation for 24 h, while more than 95% cells arrested in G2/M phase when 9NC concentration is 0. 1 μmol/L after incubation for 72 h. Apoptosis induction effect also showed a time-dependent and dose-dependent manner. Western Blot results showed the expression of Bax and Caspase-3 were upregulated while Cyclin A, Cdk2, Cyclin E and Bcl-2 were downregulated. More importantly, the compounds were more cytotoxic to the cancer cell lines than to the normal liver cell. Conclusions 9-nitrocamptothecin and 9-nitrocamptothecin liposomes can potently inhibit cell growth via regulation of cell cycle and induction of apoptosis, and this effect was preferentially in cancer cell. Inhibitory of 9-nitrocamptothecin liposomes was slightly better than the 9-nitrocamptothecin.

Objective To observe the inhibitory effect and mechanism of 9-nitrocamptothecin liposomes on HepG2 liver carcinoma cells. Methods HepG2 cells were incubated with 9-nitrocampto-thecin(9NC) or with 9-nitrocamptothecin liposomes(9NC-LP) for 24 h, 48 h and 72 h. Cell viability was then measured by the MTT assay. Cell cycle and apoptosis were evaluated by flow cytometry.Western Blot was used to determine the expression of cell cycle and apoptosis related proteins. HepG2tumor-bearing mouse models were then established. The HepG2 tumor-bearing mice were randomly divided into control group, free liposomes group, DMSO group, 9NC low dose group, 9NC high dose group, 9NC-LP low dose group and 9NC-LP high dose group. There were 10 mice in each group.Drugs were administered by tail vein and tumor volume and body weight were observed 28 days after administration. Then animals were sacrificed and the expression of proteins from tumor homogenates was analyzed by Western blotting. Results In vitro, HepG2 cell viability was apparently inhibited by 9NC and 9NC-LP, and the inhibitory effect increased in a time-dependent and dose-dependent manner.Both S and G2/M phase arrests were observed after incubation with drugs. HepG2 cells were completely arrested in S phase with 9NC concentration over than 0.1 μmol/L after incubation for 24 h,while more than 95% of cells arrested in G2/M phase when 9NC concentration was 0.1 μmol/L after incubation for 72 h. In vivo, compared with the control group, the average tumor volume was reduced in both the 9NC and 9NC-LP group (P<0.05) , and the average animal body weight also decreased in both the 9NC and 9NC-LP group (P<0.05). There was no significant difference among the control group, free liposomes group, and DMSO group. The lights inhibition rates of tumor growth in the 9NC-LP(2.5 mg/kg),9NC-LP(1.5 mg/kg),and 9NC(1.5 mg/kg)groups were 87.02%, 51.57%and 35.47%, respectively. In the 9NC-LP(2.5 mg/kg)group, >50% of animals died 14 days after drug administration. Conclusion 9NC and 9NC-LP can inhibit HepG2 cell growth via cell cycle arrest and apoptosis induction. 9NC-LP has a more potent anti-tumor effect and fewer side effects in vivo,which means 9NC-LP is a promising compound for cancer therapy via intravenous administration.

Objective To investigate the infection of Xuebijing injection on inflammatory factor of large sized avulsion patients.Methods 70 patients being selected with large sized avulsion were randomly recruited into a treatment group(35 patients)and a control group(35 patients).The control group received traditional comprehensive treatment.On this basis,Xucbijing injection was injected to the treatment group on admission day,and Xuebijing injection 50 ml in 0.9%NaCl solution 100 ml was,intravenously infused for 60 min once,2 times daily,up to 7 d.TNF-α,IL-6,CRP,WBC and NEU%of the two groups were respectively detected before treatment and 7 days after the treatment.Exudation of wound surface was also observed.Results After 7 days of treatment,there was significant difierence in the treatment group compared with pretreatment(P＜0.01).There was statistical difference between two groups after 7 days of treatment(P＜0.01).Exudation of wound surface of the treatment group was less than the control group's.Conclusion Xuebijing injection has antagonistic effect on inflammatory factor of large sized avulsion patients and can lessen exudation of wound surface.

Objective To explore the clinical efficacy of hepatic segmentectomy under segmental staining and intraoperative chemoembolization for primary liver cancer(PLC).Methods Twenty cases of liver cancer underwent hepatic segmentectomy under segmental staining and intraoperative chemoembolization(observed group),the results were compared with 22 cases of PLC after treated by routine hepatectomy(control group).AFP,CT and MRI were regularly used after hepatectomy to evaluate the outcome.Results In observed group,the operative blood loss was(295?105)mL,blood transfusion was(280?85)mL,liver function levels were in the normal range accounted for 15%(3/20) one week postoperatively,the incidence of postoperative complications was 40%(8/20),the postoperative 3-year survival rate was 60%,and the postoperative local recurrence rate was 35%;while in the control group,these parameters were(490?140)mL,(370?105)mL,40.9%(9/22),45.5%(10/22),40.91% and 68.18% respectively.In observed group,the operative blood loss,blood transfusion,cases with liver function levels in the normal range,the incidence of postoperative complications,postoperative 3-year survival rate,and postoperative local recurrence rate were significantly lower than those in the control group(P0.05).Conclusions The hepatic segmentectomy under segmental staining and intraoperative chemoembolization for PLC may reduce postoperative complications,lower postoperative relapse rate and improve survival rate.

Objective To study the effects of carboxymethylated chitosan (CMCS) to nitric oxide (NO)-induced apoptosis on rat chondrocytes,and explore p38MAPK signal transduction pathway in the process and its mechanism.Methods The rat articular cartilage cells were cultured in vitro,collagen type-2 (collagen-2) immunohistochemical staining was used to identify the cartilage cells.The model of chondrocyte apoptosis was built by different concentrations of sodium nitroprusside (SNP) induction.The cells were divided into the control group,the SNP treated group SNP+CMCS treated group,and the SNP+p38 MAPK inhibitor SB203580 treated group.The apoptotic rate of chondrocytes was calculated by FCM,apoptotic nuclei was identified by Hoechst33342 stain,the mitochondrial membrane potential changes was detected by Rhodamine123 (Rho123) stain,the expression of p38 and p-p38 were detected by Western blotting analysis.Results 1-3 mmol/L SNP could induce chondrocyte apoptosis,the apoptotic rate was increased with the SNP increasing,the most obvious apoptosis was occurred in 3 mmol/L SNP treated chondrocytes,which was 69.8％ (P＜0.05).SNP could increase the nuclear fragmentation of chondrocytes,the cells with nuclear fragmentation was significantly higher than that in the control group.SNP could reduce mitochondrial membrane potential in chondrocytes,which decreased significantly compared with the control group.SNP could increase the p-p38 expression in chondrocytes,which was 4.3 times compared to the control group.CMCS of different concentrations could reduce the apoptotic rate of SNP-induced chondrocytes,which was 51.0％,29.9％ and 15.2％,which was decreased significantly (P＜0.05) when compared with 3 mmol/L SNP induced group,CMCS decreased the cells number of SNP-induced nuclear fragmentation.CMCS increased the mitochondrial membrane potential in SNP-induced chondrocytes.CMCS reduced the expression levels of p-p38 in SNP-induced chondrocytes.Conclusion CMCS has protective effect on SNP-induced apoptosis of chondrocytes.This process is completed by inhibiting the activity of p38 MAPK signal pathway.