OBJECTIVE:To establish a HPLC method for the determination of stavudine and related substances METHODS:The Hypersil C18(4 6mm?150mm,5?m)was used as analysis column The mobile phase was methanol-water(13∶87,V/V) Detection wavelength was 264nm The flow rate was 1ml/min The column temperature was 20℃ RESULTS:The calibration curves was linear in the concentration range of 0 025～0 3mg/ml(r=0 9 999,n=6) The average recovery was 99 26%(RSD=0 76%) The contents of stavudine in 3 batches of semifinished materials and stavudine capsules(imported and domestic)were 99 48%,99 55%,99 32%(materials);97 21%,101 54%,98 92%(imported stavudine capsule);100 57%,97 86%,102 33%(domestic stavudine capsule)respectively CONCLUSION:The method is simple,accurate,sensitive and repeatable and it is suitable for the determination of stavudine and its preparations

Purpose To Prepare anti- HIV-1 gp120 monoclonal antibodies and to identify the specificity of antibodies in order to provide technique for preparing HIV remedial antibodies. Methods The gene fragment of HIV-1 gp120 was connected to PEGX-4T-2 prokaryotic expressing vector. The vector was cut by enzyme. GST-HIV protein was expressed by E. coli XL1-blue. The BALB/c mice were immunized with purified GST-HIV antigen, and then the fusion of mice spleen cell and myeloma cell SP2/0 was executed as the routine cell-fusion technique. Positive cells were screened by Indirect ELISA. Immuno-blotting assay and Western blot identified the specificity of antibodies. Results External gene section from the recombinant plasmid by sequencing showed the same size of HIV-1 gp120 gene sequences. An external expressed protein band of 32 KD was obtained after purified protein SDS-PAGE electrophoresis. It indicated that six strains of hybridoma cells secreting special monoclone antibodies had been obtained. ELISA results showed that six strains monoclonal antibodies only reacted to HIV-1 gp120 antigen. Western blot results showed that a band with molecular weight 32 kDa was obtained, which could interact with HIV-1 gp120 monoclonal antibody. Conclusion Six strains of hybridoma cells secreting special monoclone antibodies had been obtained. The prepared monoclonal antibodies have established a basis for HIV remedial antibody.

purpose To construct the expression vector of Hexastatin gene,to express and to purify the recombinant protein for further activity research.Methods The human Hexastatin gene was isolated by RT-PCR from EC9706 cells total RNA and cloned into pMDl8-T for sequencing.Then the Hexastatin gene was subcloned into pMAL-c4x expression vector and induced to express by IPTG.The recombinant fusion protein was purified with Amylose Resin Heads.Results RT-PCR product was about 687 bp and its sequence was the same as that of Hexastatin reported.The recombinant protein was expressed in E.coli BL21 with high level and the soluble protein accounted for 24.8% of the total bacterial protein,The purification of recombinant protein purified with Amylose Resin Heads reached more than 90%.Conclusion The cloning,expression and purification of human Hexastatin have laid a foundation for its anti-angiogenesis therapy for tumor.

Objective To study the endoscopic manifestations of moderate or severe chronic atrophic gastritis (CAG) related to their pathologic diagnosis.Methods The correlation analysis was done between gastroscopic manifestations and pathologic diagnosis of 49 patients with moderate to severe atrophic gastritis.Results Pathologic diagnosis of moderate to severe CAG and mild CAG were conducted in 33 and 10 cases respectively, among them, 27 cases with intestinal metaplasia (IM) and 30 cases with atypical hyperplasia. For moderate to severe CAG under gastroscopy compared with their pathologic diagnosis, the coincidence rate was 67 35%; 20 cases out of them, stained in vivo with methylene blue under gastroscopy, the coincidence rate of diagnosis was 80%. Compared with pathologic diagnosis, the gastroscopic manifestations of gastric mucosa have positive prediction rate more than 80%, when several alterations coexisted, positive prediction rate raised to more than 90%, but for every individual alteration, incidence of positive prediction was not significant. Both sensitivity and specificity of each manifestation under endoscopy were more than 95%. Other manifestations such as gastric mucosa thinning, their positive prediction rates of IM and atypical hyperplasia were less than 70%; their sensitivity and specificity were less than 30%. In using coarse and uneven lumped mucosa as the criteria in diagnosing IM and atypical hyperplasia, the sensitivity was 92 85% and 83 33% respectively, and the specificity was the same (about 70%).Conclusion Coincidence rate of diagnosis under gastroscopy and pathology is quite low in moderate to severe CAG, but the knowledge of gastroscopic manifestation and the staining technique might raise this coincidence rate.

Objective To investigate the influence of kaempferol on transforming growth factor(TGF)-β1/Smads signal trans-duction in liver tissue of mice with schistosomiasis liver fibrosis. Methods Forty BALB/c mice were randomly divided into a normal control group(8 mice),a praziquantel group(8 mice ),and 4 praziquantel+kaempferol groups with different kaempfer-ol dosages(5,10,15,20 mg/kg respectively,6 mice each group). Besides the normal control group,all the mice in the other 5 groups were infected with Schistosoma japonicum. After the infection for 6 weeks,the praziquantel group and the 4 praziquantel+ kaempferol groups were treated with praziquantel 500 mg/(kg · d) for 2 d,then the mice in the praziquantel group were drenched with normal saline for 6 weeks,and those in the 4 praziquantel+kaempferol groups were drenched with kaempferol 5, 10,15,20 mg/kg respectively for 6 weeks. After the treatment,all the animals were sacrificed by the cervical dislocation meth-od,and the area of egg granuloma and the degree of fibrosis in the livers of the mice were observed by HE and Masson staining. The expressions of TGF-β1,Smad2/3,Smad7 proteins were measured by the immunohistochemical method,and the mRNA lev-els of the 3 proteins were detected by RT-PCR. Results Compared with the mice in the praziquantel group,the areas of egg granuloma of the liver of the mice in the 4 praziquantel+kaempferol groups were smaller,and the degrees of the hepatic fibrosis of the mice were lesser,and their expressions of Smad2 and Smad3 at protein and their mRNA levels were significantly lower (all P<0.05),while the expression of Smad7 at protein and its mRNA level were significantly higher(all P<0.05). Conclu-sion By decreasing the expressions of TGF-β1 and Smad2/3,and increasing the expression of Smad7,kaempferol can signifi-cantly reduce the degrees of hepatic fibrosis and granuloma caused by schistosome eggs after the praziquantel treatment.

OBJECTIVE:To prepare famotidine sodium chloride injection METHODS:The famotidine sodium chloride injection was prepared with aspartic acid as solutizing agent and sodium chloride injection as solvent The contents of famotidine and its related substances were determined by means of HPLC Influencing factor and the stability of famotidine were studied and the therapeutic effect was observed RESULTS:The content of famotidine was slowly reduced while temperature rising and time prolongine The content of famotidine was 98 6% at the end of the 6th month at 40℃ with accelerating experiment Its content was 98 4% after one year storage under room temperature Its content was 98 5% at the 10th day after illumination with 4 500Lx light in accelerating experiment The content of related substances was less than 2% The total effective rate was 93 75% CONCLUSION:Famotidine sodium chloride injection was stable in property and had definite therapeutic effect,and its clinical application was simple and free from contamination

OBJECTIVE:To prepare captopril hydrophilic gel sustained-release tablet and study the influencing factors on its release METHODS:With HPMC as the matrix,the tablets were prepared by direct compression method and influencing factors on release were studied RESULTS & CONCLUSION:The in vitro release of prepared tablets conformed to Higuchi equation The HPMC matrix tablet could release in a sustained way when the proportion of HPMC was at least 15% in weight and the best proportion was 60%;The dissolution of Methocel K was slower than 60RT or 75RT;Taking lactose as the filler was better than starch or CaSO4;When the proportion of lactose increased,the dissolution sharply decreased;The tablet dissolved more rapidly with paddle method than with rotating basket methos,compression force and pH of dissolution medium affected the release very little

ObjectiveTo investigate the value of a digital three-dimensional reconstruction technique in the treatment of hepatic alveolar echinococcosis (HAE).MethodsThe computed tomography scan data for 13 patients with HAE who were admitted to the First Affiliated Hospital of Xinjiang Medical University from February 2011 to October 2011 were reconstructed and analyzed by a three-dimensional reconstruction system to assess resectability,and to facilitate surgical planning and individualized virtual surgery.The results of preoperative analysis were compared with the results of actual operations.ResultsThe three-dimensional models of the liver were reconstructed successfully,and intrahepatie lesions and vessels were clearly displayed.One patient received an autologous liver transplantation,10 underwent hepatectomy,and 2 received percutaneous transhepatic cholangial drainage.Virtual operation planning was carried out for 11 patients using the three-dimensional reconstruction system.The mean volume of the liver to be resected was predicted to be 920 ml (range,339-2678 ml),and the mean percentage of liver to be resected to the total liver volume was predicted to be 45％ ( range,23％ -68％ ).The mean volume o[ the actual liver resection was 834 ml (range,315-2250 m[),and the mean percentage of actual liver resected to the total liver volume was 42％ (range,22％ -70％ ),which was consistent with the results of preoperative three-dimensional reconstruction.All patients were followed up for 2-8 months,and no severe complications such as liver failure,hemorrhage and bile leakage were detected.ConclusionDigital three-dimen-sional reconstruction is helpful in the diagnosis and treatment of HAE and effectively reduces surgical risks.

Objective To investigate the effects of lentiviral-mediated RNA interference (RNAi) targeting tumor necrosis factor-α(TNF)-αgene on the expression of TNF-α, interleukin (IL)-1β, IL-6 of murine macrophages RAW264.7, and the efficiency of RNAi experimental gene therapy for the murine collagen-induced arthritis (CIA). Methods The RAW264.7 macrophages were infected by lentivirus-RNAi particles, then stimulated by Lipopolysaccharides (LPS). The TNF-α, IL-1β, IL-6 expression of RAW264.7 macrophages were measured with real-time polymerase chain reaction (PCR) and enzyme linked immunosorbent assay (ELISA). CIA models were esta-blished in DBA/1 mice using bovine type Ⅱ collagen. The treatment effect of lentivirus-RNAi on CIA were observed through arthritis scores, serum TNF-α measurement and hind paw paraffin section hematoxylin/eosin staining after lentivirus-RNAi particles tail vein injection. Results The TNF-αmRNA relative expression level of lentiviral RNAi group was 0.291 ±0.021, significantly lower than that of negative control group 0.925±0.013 (t=25.4, P<0.01). The inhibition rate in mRNA levels was 68.5%. The serum TNF-α level of lentiviral RNAi group was [(249 ±11) ng/ml], significantly lower than that of negative control [(382±6) ng/ml] (t=10.31, P<0.05). The inhibition rate of protein levels was 34.7%. It had no effect on the IL-1β and IL-6 mRNA expression. On the 8th day after systemic administration, the arthritis score of lentivirus-RNAi group was 2.50±0.19, which was significantly lower than that of blank controls (3.63 ±0.18) and negative controls (3.75 ±0.16) (F=42.8, P<0.01). From now on, arthritis score of lentivirus-RNAi group and positive control decreased slowly to at least 2 weeks after treatment induction. The serum TNF-α levels of lentivirus-RNAi group and positive controls were [(35±6) pg/ml] and [(32±7) pg/ml] significantly lower than that of negative controls [(47±3) pg/ml] (t=3.03, 4.11, P<0.01) respectively. Morphological examination showed that the lentivirus-RNAi decreased CIA pathological manifestations. Conclusion Lentiviral-mediated RNAi targeting murine TNF-α gene can effectively inhibit TNF-α expression both in vitro and in vivo, which also effectively improve the CIA arthritis score. Lentiviral-mediated RNAi targeting TNF-αgene provides a potential strategy for rheumatoid arthritis (RA) treatment.

Objective To evaluate the efficacy and safety of adefovir dipivoxil(ADV)in treating patients with hepatitis B e antigen(HBeAg)positive chronic hepatitis B.Methods In this randomized,double blind,placebo-controlled,multicenter trial,210 eligible patients with HBeAg positive chronic hepatitis B were recruited and randomized(randomization ratio was 2:1)receiving ADV 10 mg/d for 48 weeks(ADV+ADV group,n=142)or placebo for 24 weeks followed by ADV 10 mg/d for 24 weeks(PLB+ADV group,n=68).The primary endpoint was virological response. The secondary endpoint was serologic response(HBeAg loss rate and HBeAg seroconversion rate) and alanine aminotransferase normalization rate.Results After 24 weeks therapy,mean reduction of hepatitis B virus(HBV)DNA level comparing with that of baseline was 3.12 log_(10)copy/mL in ADV +ADV group while it was 0.95 log_(10)copy/mL in PLB+ADV group.The percentages of patients with HBV DNA clearance(HBV DNA level