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 .

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.

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.

BACKGROUND:It has been confirmed that carboxymethylated chitosan has an promoting effect on Schwann cel proliferation and secretion, but its impact on the cyclic adenosine monophosphate-mediated protein kinase A signaling pathway in schwann cel stil needs further study. OBJECTIVE:To investigate the effect of carboxymethylated chitosan on cyclic adenosine monophosphate/ protein kinase A signaling pathway in rat schwann cels. METHODS:The Schwann cels of the second generation neonatal rats were obtained and seeded in 6-wel plate at a concentration of 1×109/L. These Schwann cels were cultured and divided into four groups. The Schwann cels in the control group were cultured by adding PBS. The Schwann cels in the experimental groups were cultured by adding 50, 100 and 200 mg/L of carboxymethyl chitosan solution, respectively. After 24 hours, the concentration of cyclic adenosine monophosphate, protein kinase A activity and cyclic adenosine monophosphate response element binding protein mRNA expression were detected. RESULTS AND CONCLUSION:Compared with the control group, carboxymethyl chitosan increased cyclic adenosine monophosphate concentrations, the activity of protein kinase A and cyclic adenosine monophosphate response element binding protein mRNA expression within the Schwann cels in a dose-dependent manner (P < 0.05). These results demonstrate that carboxymethyl chitosan can increase the concentration of cyclic adenosine monophosphate within the Schwann cels and promote protein kinase A activity, thereby activating cyclic adenosine monophosphate/protein kinase A signaling pathway.

TiO2 coated hollow fiber ( HF) was prepared by sol-gel method and characterized by X-ray powder diffraction and scanning electron microscope. The adsorption performances of the self-prepared TiO2 coated HF for interest metal ions were explored. The sample solution was extracted for 30 min at pH 8. 0 under stirring at speed of 700 r/min, then desorpted with 100μL of 1 mol/L HNO3 . On the basis of this, a method combining TiO2 coated HF micro-solid phase extraction with electrothermal vaporization-inductively coupled plasma mass spectrometry ( ICP-MS ) was developed for the determination of trace metal ions in environmental water samples. Under the optimized conditions, the limits of detection obtained by the proposed method were 0. 039, 0 . 021 , 0 . 009 and 0 . 018 ng/mL for CrⅢ, CuⅡ, CdⅡ and PbⅡ with enrichment factors of 12 . 5 , 11 . 7 , 10. 3 and 18. 6, respectively. The preparation reproducibility of self-prepared TiO2 coated HF ranged from 4. 5% to 6. 8% (n=9) in one batch, and from 7. 7% to 9. 6% (n=7) in batch-to-batch. The developed method has been validated by analyzing a certified reference material ( GSBZ50009-88 200925 ) and also applied to the analysis of CrⅢ, CuⅡ, CdⅡ and PbⅡ in East Lake water and snow water successfully. Meanwhile, TiO2 coated HF stir bar was prepared for a comparison and it was demonstrated that TiO2 coated HF displayed higher extraction efficiency and adsorption capacity for target ions.

〔Abstract〕 Medical and health sci-tech novelty assessment plays a key role in information supporting in the hospital scientific research and management work of health sector.Combining with the current status of medical and health sci-tech novelty assessment work station in the Military Hospital of Beijing PLA, the paper puts forward countermeasures acoording to the existing problems, namely, weak management and personnel and so on, in order to promote comprehensive development of medical and health sci-tech novelty assessment work.