AIM: To investigate protect effect of the NO donating-oleanlic acid derivatives on hepatic fibrosis in rats. METHODS: The rat model with early hepatic fibrosis was induced by carbon tetrachloride (CCl_4). ZCⅡ_2 at the dose of 128 and 64 mg?kg -1 had been given orally for 30 days. Serum level of the total protein (TP), albumin (ALB), the albumin/globulin (A/G) and ALT, AST. The hyaluronic acid (HA), laminin (LN), the procollagen type Ⅲ (PCⅢ) and the level of MDA, GSH-Px in liver tissues were determined. Pathological examination to reveal the extent of liver damages was observed. [WTHZ]RESULTS: ZCⅡ_2 at the dose of 128 mg?kg -1 increased the serum TP, ALB, and A/G and decreased HA, LN, PCⅢ, ALT, and AST more significantly than the model group, and hepatic pathological injury was abated to some degree. CONCLUSIONS: ZCⅡ_2 at the dose of 128 mg?kg -1 can attenuate liver damages, protect against lipoperoxidation and attenuate hepatic fibrosis induced by CCl_4.

Aim To study the inhibitory effect of NO donating-aspirin derivative Ⅱ_6 on thrombosis and its mechanism of nitric oxide release. Methods Inferior vena cava thrombosis and cerebral thrombosis model in rats were used to investigate the antithrombotic activities of Ⅱ_6.TXB_2 and 6-Keto-PGF_(1?)in rat serum、rat aortic strip and in ECV304 cells were determined by radioimmunosay.NO content in the serum of rats and extracellular fluid of the ECV304 cells were tested with Griess reagent.Results Ⅱ_6 inhibited the thrombosis in inferior vena cava and brain,and it reduced the concentration of TXB_2.but increased NO production.Conclusion Ⅱ_6 has an inhibitory effect on thrombosis and increases NO content in vivo and vitro.

AIM: To estimate the antiasthmatic activity of new compounds: SDF-1, SDG-1 and SDG-3. F-1 is a furoxan derivative, G-1 and G-3 are hydroxylguanidine derivatives, SDF-1 is a novel seratrodast derivative connected with F-1, SDG-1 a seratrodast derivative connected with G-1, and SDG-3 a novel seratrodast derivative connected with G-3. METHODS: Firstly, the in vivo antiasthmatic activity was estimated in asthmatic guinea pigs induced by acetylcholine and histamine. Secondly, the in vitro NO releasement of these compounds was determined following the procedures of Griess. Finally, tracheal smooth muscle relexant potency of these compounds was evaluated on trachea of guinea pigs. RESULTS: The in vivo antiasthmatic activity of SDF-1 was more potent than seratrodast (P

Objective To exclude false positivities in detection of IgM antibodies against hepatitis E vires of genotype 4 (HEV-4) using a truncated immunodominant polypeptide of HEV open reading frames (ORF2). Methods The recombinant ORF2 immunodominant polypeptide corresponding to amino acids (AA) 459-607 and a truncated polypeptide corresponding to AA 472-607 were separately applied to coat ELISA plates. Anti-HEV IgM from 35 serum samples with HEV RNA positive, 69 serum samples from healthy individuals and 117 clinically suspicious HEV RNA positive serum samples was detected by an indirect ELISA and was confirmed by western blot in protein level and RT-PCR detecting in RNA level. Results Western blot analysis showed that the sera from HEV patients reacted with the dimmer of peptide 459-607, but they didn't react with the monomer and peptide 472-607. The ELISA showed that all 35 serum HEV RNA positive samples reacted with peptide 459-607 but not with peptide 472-607 and none of the 69 serum samples from healthy individuals reacted with either polypeptide. Among 117 chnically suspicious HEV RNA serum samples, 5 samples reacted simultaneously with both polypeptides. But the difference between 450 nm absorbance (A450) value was less than 0. 5. Western blot analysis demonstrated that all the 5 serum samples were anti-HEV IgM- negative. The 5 serum samples was detected negative by RT-PCR, indicating that the false pesitivities were caused by non-specific absorption. Conclusions ORF2 peptide 459-607 may be used to detect anti-HEV lgM efficiently. The false positivities caused by non-specific absorption can be largely excluded according to the difference between 45Ohm absorbance (A450) value when serum reacts with both polypeptides.

Objective To characterize the dimerization and the antigenicity of the ORF2 polypep-tide of hepatitis E virus (HEV, genotype 4). Methods HEV ORF2 gene was cloned from the serum of a patient with hepatitis E. The genotype was determined by sequencing. Three ORF2 polypeptides differing in size and other polypeptides with point mutations were produced in E. coli. The recombinant polypeptides were purified and analyzed by SDS-PAGE and Western blot. Results The ORF2 polypeptide containing 459-607 amino acid formed homedimer even in 8 mol/L urea. The truncated polypeptides containing amino acid 472-607 or 459-594 formed monomer only. The mutations at amino acid 562 or 595 disrupted the ho-modimer, whereas the mutations at amino acid 476 or 580 did not. Anti-HEV from hepatitis E patients only reacted with the homodimer form of the polypeptide 459-607 and did not react with monomer or tnmcated pol-ypeptides. Conclusion The amino acid 459-607 of HEV ORF2 is essential for dimerization of the ORF2 polypeptide. Residues at amino acid 562 and 595 are critical for the dimerization. The antigenicity of the polypeptide 459-607 mainly depends on its homodimer form.

Aim: To study the synthesis and anti-inflammatory activity of p-(sulfamyl) benzylidene-linked hetero-cyclic ketone derivatives. Methods: A series of p-(sulfamyl) benzylidene-linked heterocyclic ketone derivatives were synthesized. Anti-inflammatory activity was evaluated against xylene-induced ear oedema in mice and against carrageenan-induced paw oedema in rats. Gastrointestinal side effects in the rats were also examined after continu-ous introgastric administration of these compounds once daily for 7 days. Results: Twelve compounds( LHZ-101-LHZ-112) were synthesized and their structures were confirmed by IR, ~1H NMR, MS and elemental analysis. LHZ-105, LHZ-106 and LHZ-111 exhibited marked anti-inflammatory activity in xylene-induced mice ear swelling model. LHZ-106 and LHZ-111 showed significant anti-inflammatory activity in carrageenan-induced rat paw ede-ma model. LHZ-105, LHZ-106 and LHZ-111 had less gastrointestinal side effects than diclofenac sodium and CI-1004. Conclusion: These results suggest that some of these compounds have the potential for anti-inflammatory activity with few gastrointestinal side effects.

Objective To obtain highly purified astrocytes and identify the cells in each stage to support further studies.Methods The cerebral cortex of a neonatal SD rat was isolated and prepared into single cell suspension.The obtained cells were purified by differential adherence and shook at a constant temperature.By inverted phase contrast microscopy and HE staining,cell morphology was observed.The immunofluorescence staining with anti-mouse GFAP was used to identify the cells.Results The primary cortical cells developed rapidly at 3 d after culture and covered the flasks at 9-12 d.At this time,the cells showed stratification and the astrocytes lay at the lower layer.GFAP positive rate was only about (67.2 ±7.1)%.After the first passage,GFAP positive rate increased obviously (84.0±6.0)%. However, oligodendrocytes and microglias could not be removed completely,and the cells also showed stratification.Through 3 times of passages,we obtained many single species of astrocytes showing satellite shape with 2 or 3 processes,big cell body and round or oval-shaped nuclei leaned to one side.Immunofluorescence staining showed that nearly all of the cells were strong positive and the positive rate reached as high as (97.6 ± 2.4 )%.Conclusion Through differential adherence and shaking at a constant temperature,more astrocytes of high purity and in good state can be obtained.

Background and purpose:Drug resistance is a major cause of failure in lung cancer chemotherapy. This study aimed to investigate the effect of YAP on doxorubicin resistance in lung cancer and its underlying mechanism. Methods:Doxorubicin resistant lung cancer cell clones were established from parental sensitive cancer PC9 cell line via in vitroinduction, and the expression of YAP was analyzed. YAP was down-regulated via shRNA to different levels. MTS assay was employed to determine cell proliferation and drug sensitivity. Flow cytometry was used to determine cell cycle distribution, apoptosis and uptake of Rh-123. Western blot and quantitative real-time polymerase chain reaction (QRT-PCR) assay were used to determine the expression of ABCB1, ABCC1, p53, Runx2, ITGB2 and ErbB4. The phosphory-lation of serine/threonine kinase (AKT) was determined as well.Results:Doxorubicin resistant PC9/Adr cell clone was obtained with over-expressed YAP. Expression of YAP in PC9/Adr cells was down-regulated to different levels via shR-NA. After YAP silencing, cell proliferation was reduced, while sensitivity to doxorubicin was increased. The cell cycle was significantly halted by G0/G1 phase. Doxorubicin induced-apoptotic rate and cellular uptake of Rh-123 were increased,with positive correlation to YAP silencing level. Western blot and QRT-PCR results showed that after YAP silencing, ABCB1, ABCC1, Runx2, ITGB2, and ErbB4 proteins were down-regulated, while the expression of p53 was up-regulated. Phosphorylation of AKT was inhibited as well.Conclusion:Over-expression of YAP is involved in doxorubicin resistance in PC9/Adr cell line. Silencing of YAP could restore doxorubicin sensitivity. The mechanism involves regulation of drug resistance-related genes and promotion of apoptosis.

Phthaloyl dichloride (1)was reacted with LiAlSeH2 to give benzo[c]selenophene-1;3-dione (2);which was treated with the Grignard reagents to generate hydroxyl compounds 3a-3h.These compounds were finally converted to target products 4a-4h by treatment with hydriodic acid.The structures of 4a-4h were confirmed by MS and 1 H NMR.Their inhibitory activity against adenosine diphosphate (ADP)-induced platelet aggregation was evaluated by Born′s turbidimetric assay;free radical scavenging activity was assayed by xanthine oxidasemethod and 1;10-phenanthroline spectrophotometric method.It was found that compound 4 f displayed more potent inhibi-tory effect on platelet aggregation than 3-n-butylphthalide and comparable hydroxyl free radical scavenging activity in vitro to that of edaravone.Therefore;compound 4 f might be the candidate for further investigation.

Objective To study the clinical pathological features,treatment and prognosis of umbilical metastases of intra abdominal malignancies (sister Mary Joseph nodule, SMJN). Methods From January 1980 to July 2003, 17 SMJN cases were admitted. The clinical features were reviewed. Results The diagnosis of SMJN was confirmed by pathology in all cases, including epithelial ovarian carcinoma in 6 cases, gastric adenocarcinoma in 6 cases,colon adenocarcinoma in 2 cases, and endometrial carcinoma, gallbladder carcinoma and undifferentiated adenocarcinoma without identifying primary site in one each. Mean survival、half year、one year and two year survival rate in 10 palliative excision cases was 13 5 months、60%、20% and 10%, which were significantly better than 8 25 months、14 3%、0% and 0% in 7 conservative therapy cases ( P