OBJECTIVE To set up a kind of Helicobacter pylori(Hp) detection method which can determine the Hp infection and its clinical value for the digestive diseases.METHODS Gastric biopsy was used with gastric mucosa Hp rapid urease test,Hp-IgG was detected by ELISA,and Hp-DNA of feces by fluorescent gene quantitation method.RESULTS From the 96 patients positively determined by gastric biopsy,the positive rate of Hp-DNA of feces was 95.8%,but the positive rate of Hp-IgG was 47.9%.From the 59 Hp-IgG positive patients,the positive rate of gastric biopsy was 45.8%.CONCLUSIONS Gastric biopsy and HP-DNA quantitation of feces are both reliable methods which can determine the Hp infection,but Hp-DNA quantitation of feces is a simple,rapid,reliable and atraumatic test,it can be put into practice widely and more easily.

Aim To investigate the protective effect of catechin on cerebral ischemia-reperfusion injury in rats and its mechanism.Methods 40 rats were randomly divided into 5 groups:sham operation group,model group and 50,100 and 200 mg·kg~(-1) catechin groups,with 8 rats in each group.The model of focal cerebral ischemia-reperfusion in rats was established with modified sutured-occluded method.The rats in catechin groups were injected with catechin at the matched concentration.The rats in sham operation group and model group were injected with saline.And all rats were given more time in 2 hours after ischemia.Rats were sacrificed for histologic examination after the behavioral test,and their brains were taken to assay the activities of MPO and NOS.Results Catechin at different dosages(50,100 and 200 mg·kg~(-1))could obviously decrease neurological deficit score,repair histological injury,and reduce the activities of MPO and NOS in rats of focal cerebral ischemia-reperfusion injury.Conclusions Catechin can relieve the cerebral ischemia reperfusion injury,and its mechanism may be partly related to the effects of its antiinflammation and antioxidation.

Objective To investigate the cross reaction characteristics of recombinant human papillomavirus 16 type L2 full-length fusion protein in HPV types of 6, 11, 18.Methods The serum samples of 108 condyloma acuminatum patients, 156 cervix cancer patients and 100 healthy control subjects were collected.The gene of full-length HPV16 L2 was amplificated from the tissue DNA of cervical cancer patient and inserted into the prokaryotic expression vector PGEX-4T-1 to construct the recombinant plasmid PGEX-4T-1-HPV16 L2.After sequencing identification, the recombinant plasmid was tranformed into E.coli BL21 (DE3).After induction by IPTG, the fusion protein containing HPV16 L2 was expressed and analyzed by both SDS-PAGE and WB.Furthermore, the specific binding capacity of the fusion protein to the HPV 6,11, 16 and 18 DNA positive patient's sera were analyzed by WB.The fusion protein was purified with NiNTA Agarose Kit and coated with ELISA reaction plates.The specific serum IgG were detected by indirect ELISA.Results The recombinant plasmid PGEX-4T-1-HPV16 L2 was constructed successfully. Highly expressed HPV16 L2 full-length fusion protein was obtained and the expression level was 27.2 %.The relative molecular mass(Mr) of the fusion protein is about 82 × 103, which matches up to the expected Mr.Meanwhile, the sera of HPV 6,11,16,18 DNA positive patients were used as the primary antibody and the Mr of the specific band was detected to be about 82 × 103 by WB.The results of indirect ELISA showed that the average levels of specific IgG in condyloma acuminatum group, cervical cancer group and healthy control subjects were 0.848 ±0.257, 0.822 ±0.247 and 0.173 ±0.143 with the positive rate of 92.6%, 94.2%and 8.0% respectively.There was no significance of the specific IgG levels between condyloma acuminatum group and cervical cancer group ( F = 0, P ＞ 0.05 ), but there was significant difference of specific IgG levels and positivity among the three groups ( F = 305.201 ,x2 = 253.178, P ＜ 0.01 ).Conclusions The HPV16 L2 full-length fusion protein has better antigenicity.However cross reactions with HPV6, 11 and 18 were found.It can be applied in serological screening reagents for HPV infection and associated cancer.

Objective To express in prokaryotic system and to analyze the antigenic specificity of EB virus(EBV) latent membrane protein 2(LMP2) multi-epitopes gene rich of T cell and B cell epitopes.Methods Using on-line prediction service, T cell epitopes and B cell epitopes of EB virus latent membrane protein 2 were predicted. The genes rich of CTL and th cell epitopes were selected as the candidate gene sequences, while B cell epitopes around them were taken into account. The finial selected multi-epitope gene was synthesized after being optimized according to prokaryotic codon bias and inserted into prokaryotic expression vector pET32a( + ) to get the recombinant plasmid: pET32a( + )/EBV-LMP2 multi-epitopes. After transformed into E. coli BL21 (DE3) and induced by IPTG, the target multi-epitopes gene can be expressed as Trx-His fusion protein. The expression products can be identified by SDS-PAGE and Western blot. Moreover, rabbit serum antibody to EBV membrane protein and nasopharyngeal carcinoma(NPC) patient serum were used respectively to detect the antigenic specificity of the multi-epitopes. Meanwhile, 6-8 weeks female BALB/c mice were immunized with EBV-LMP2 multi-epitope at 2 week intervals, three times in all, Trx-His protein and PBS were set as the control groups. At the second week after the last immunization, the mice were sacrificed. LDH and indirect ELISA were taken to detect the specific spleen CTL activityand specific IgG in serum, which reflected the immunogenicity of the EBV-LMP2 multi-epitope. Results Two amino acid sequences which locate at the LMP2 (aa195 -232 ) and LMP2 (aa419-436 ) were selected and connected in series to be the target gene. The recombinant plasmid containing EBV-LMP2 multi-epitope gene successfully constructed and the target protein was expressed in E. coli BL21 ( DE3 ). The relative molecular mass(Mr) of The expression products is about 27 × 103 , which matches up to the expected Mr. The antigenic specificity of the multi-epitopes protein was identified by Western blot and the multi-epitopes protein also can be detected by rabbit serum antibody to EBV membrane protein and NPC patient serum respectively. In the result of the animal experiment, EBV-LMP2 multi-epitope was able to induce the specific CTL activity in BALB/c mice. With the increasing of the effector: target ( E: T) 1: 5,1: 10, 1: 25, the CTL activity was also increased wih( 12.52% + 2.59% ), (21.80% + 1.08% ), (23.68% + 3.74% ) respectively; EBV-LMP2 multi-epitope was able to induce LMP2-specific antibody response(A490 =0.258 +0.040) as compared with Trx-His protein(A490 =0.095 +0.011) and PBS(A490 =0.068 +0.014,P＜0.05＝. Conclusion The EBV-LMP2 multi-epitopes gene was designed successfully and expressed precisely in prokaryotic expression system with good antigenicity and immunogenicity.

ObjectiveTo study the anti-cancer effects and mechanism of Gexia-Zhuyu decoction combined with cyclophosphamide on EAC liver cancer mice.MethodsChoose EAC liver cancer mice cell lines and production of animal models of tumor.The mice were completely randomized into blank group,model group,the Chinese medicine group,CTX group,Chinese medicine and CTX group.The blank group and model group were given normal saline 0.4 ml/d; traditional Chinese medicine group was administrated with Gexia-Zhuyu decoction (11 g/kg body weight ) 0.4 ml/d gavage; CTX group was given intraperitoneal injection of 2 mg/ml of cyclophosphamide 0.4 ml/d; and traditional Chinese medicine and CTX group was given Gexia Zhuyu decoction 0.4 ml/d orally and CTX intraperitoneal injection of 0.4 ml/d.Ten days after the administration,the body weight of mice and the general condition were tested,and morphology was observed under the light microscope electron microscope mesenchymal tumor angiogenesis situation.Results Gexia-Zhuyu decoction combined with cyclophosphamide in mice could improve appetite,gain weight,and greatly reduce the tumor stroma in the number of capillaries under the electron microscope together with significant pathological morphological changes.Conclusion Gexia-Zhuyu decoction combined with cyclophosphamide had anti-cancer effects which can improve the quality of life and inhibit the growth of tumor in interstitial capillaries.