Objective: To study the isolation, identification, and biological characteristics of the secondary metabolites of myxobacteria. Methods: A growth inhibition model of Pyricularia oryzae was used to screen the active microbes. The compounds extracted with methanol from the fermentation broth of Myxococcus xanthus 095B06 were separated by silica chromatography, gel filtration chromatography, and high-performance liquid chromatography. The chemical structures of the compounds were identified by 1HNMR, 13CNMR, ESI-MS, and EI-MS techniques. The bioactive activities of the separated compounds were evaluated by Kirby-Bauer Disc Diffusion method and MTT method. Results: Six compounds, namely, Avermectin Ala, Avermectin A2a, Avermectin Bla, Avermectin B2a, ergosta-7,22-dien-3,5,6-triol, and 4-quinolinecarboxylic acid, were obtained. Conclusion: Compound 1, 2, 3, 4, and 5 have been isolated from myxobacteria for the first time and compound 2 and 4 can strongly inhibit the growth of SMMC-7721 cell line, with both IC50 values being 5 g/ml.

Adrenergic beta-Antagonists ; therapeutic use ; Drug Therapy, Combination ; Esophageal and Gastric Varices ; drug therapy ; Humans ; Isosorbide Dinitrate ; analogs & derivatives ; therapeutic use

Objective To test the hypothese that extreme low estrogen level will lead to elevation of blood pressure.Methods Thirty-two adult male SD rats were submitted to following approaches:control(n=8),orchi- ectomy(n=8),orchieetomy plus letrozole,an inhibitor of estrogen synthese [5 mg/(kg?d)by gavage,n=8], letrozole treatment in intact testis rats(n=8).Four weeks after treatment,SBP,serum testosterone(T),estradiol (E_2),nitric oxide(NO),endotheline(ET),thromboxane(TXB_2),prostacyelin(6-Keto-PGF_(1?)),atrial naturetic pep- tide(ANP),C type natriuretic peptide(CNP)were determined.Results SBP was increased significantly after or- chiectomy compared with control rats(orehiectomy:171?17 vs control:156?14 mmHg,P

Objective To investigate the method and value of adjustable interatrial fistulization in the operation of congenital heart disease(CHD) accompany with severe pulmonary arterial hypertension(PH).Methods Twenty-seven patients(19 male,8 females) accompany with severe PH were entered the study,age ranged from 4 to 14 years old,weight from 13.7 to 42.0 kilogram.The enrolled diseases included 11 cases of atrial septal defect(ASD),10 cases of ventricular septal defect(VSD),4 cases of patent ductus arteriosus(PDA),and 2 cases of Ebstein syndrome accompany with severe tricuspid insufficiency.All patients were diagnosed as CHD accompany with severe PH(bidirectional shunt)which was the contraindications for routine operation before operation through chest X-ray,electrocardiography,ultrasonic cardiography,cardiac catheteri-zation and cardiac angiography.Results With adjustable interatrial fistulization and treatment to the abnormalities,14 fistulaes were closed immediately after operation,7 fistulaes were closed 2 days after operation,3 fistulaes were closed 3 days and 1 fistulae was closed 4 days after operation and accompanied with empyema discharged initiatively.One fistula was never closed,1 case died from low cardiac output symptom.The effective rate was 92.6%,closed to that of routine operations.Conclusion Adjustable interatrial fistulization is an easy procedure,and it can decrease the danger of PH post-operation effectively and provide operation opportunity for those patients with CHD approaching terminal stage.

This study is to observe the effect of ilexonin A (IA) on the expression of basic fibroblast growth factor (bFGF) and growth associated protein-43 (GAP-43), and neurogenesis after cerebral ischemia-reperfusion in rats and explore its possible mechanism of protecting neuronal injury. Models of middle cerebral artery occlusion (MCAO) were established in SD rats. Before and after two hours ischemia-reperfusion, IA (20 and 40 mg x kg(-1)) was injected immediately and on 3, 7, 14, and 28 d once a day. The neurological severity was evaluated by neurological severity scores (NSS); neuronal injury in the boundary zone of the infarction area was evaluated by TUNEL and Niss1 staining. The expressions of bFGF and GAP-43 and neurogenesis were evaluated by Western blotting and 5-bromodeoxyuridine (Brdu) fluorescence staining, respectively. After treatment with IA, the NSS of treatment groups were lower than that of the models (3 and 7 d). The number of TUNEL positive neurons decreased and Nissl positive neurons increased at the same time (3 d). The expressions of bFGF and GAP-43 increased significantly in the boundary zone of the infarction area when compared to model group. Moreover, IA markedly enhanced the neurogenesis in the brain after ischemia-reperfusion, which revealed an increase of Brdu/NeuN positive cells in the boundary zone of the infarction area. The possible mechanism of protecting neuronal injury of IA may be related to inhibition on neuronal apoptosis, upregulation of bFGF and GAP-43, and neurogenesis in boundary zone of infarction after cerebral ischemia-reperfusion.
Animals ; Apoptosis ; drug effects ; Brain Ischemia ; etiology ; Bromodeoxyuridine ; metabolism ; Fibroblast Growth Factor 2 ; metabolism ; GAP-43 Protein ; metabolism ; Infarction, Middle Cerebral Artery ; complications ; Male ; Neurogenesis ; drug effects ; Neurons ; pathology ; Neuroprotective Agents ; pharmacology ; Organic Chemicals ; pharmacology ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; etiology ; metabolism

OBJECTIVEThe purpose of this study is to study the sealing ability and the furcal appearance of repairing subpulpal wall perforation with resinous inlay.

METHODSFifty newly extracted human molars were randomly divided into three experiment groups (group A, group B, group C, 15 teeth each) and one control group (5 teeth). In experiment groups, perforations were made perpendicularly to the center of the pulp chamber floor. Perforations of group A and B were repaired with resinous inlay and sealed by AH Plus sealer and luting glass-ionomer, respectively. Perforations of group C were directly repaired using light-cure composite resin. Perforations were not made in five teeth of control group. The furcal appearances were evaluated under stereomicroscope after repairing. Microleakage was measured by glucose oxidase detection.

RESULTSThe fineness rate of furcal appearances with resinous inlay repairing were 83.3%, while the fineness rate of furcal appearances with light-cure composite resin directly repairing were 46.7%. There were statistics difference between resinous inlay repairing and light-cure composite resin directly repairing (P<0.05). There were statistics difference among the daily microleakage of three experiment groups, group A

CONCLUSIONUsing resinous inlay to repair the subpulpal wall perforation can improve the sealing effect and avoid material overextension. AH Plus can be used as perforation sealant because of its better sealing ability.

Bicuspid ; Composite Resins ; Dental Leakage ; Dental Pulp Cavity ; Glass Ionomer Cements ; Humans ; Inlays ; Molar

OBJECTIVETo explore the role of estrogen in the regulation of the expression of claudin-6 and biological behavior in MCF-7 cells.

METHODSRT-PCR and immunocytochemistry were conducted to analyze the expression and localization of claudin-6 in MCF-7 cells treated with 17β-estradiol. CCK-8 kit assay and Scratch Test were conducted to analyze the capability of proliferation and migration of 17β-estradiol treated MCF-7 cells.

RESULTSRT-PCR analysis and immunocytochemistry showed that 17β-estradiol induced a concentration-and time-dependent effect on claudin-6. At 5 nmol/L and at 24 h, 17β-estradiol treatment led to an increased level of claudin-6, which was located in the membrane of MCF-7 cells. CCK-8 analysis showed a significant decrease in the capability of proliferation of MCF-7 cells compared with the control group (P < 0.05). Cells Scratch Test showed decreased migration capability of MCF-7 cells compared with the control group (P > 0.05).

CONCLUSIONS17β-E2 might regulate the expression of claudin-6 and inhibit the proliferation and migration of MCF-7 cells. The inhibitory effects of 17β-E2 on growth and migration of MCF-7 cells may be mediated by claudin-6 expression regulation.

Breast Neoplasms ; metabolism ; pathology ; Cell Line, Tumor ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Claudins ; Dose-Response Relationship, Drug ; Estradiol ; administration & dosage ; pharmacology ; Female ; Humans ; Membrane Proteins ; metabolism

OBJECTIVETo observe the toxicity of Pulsatilla chinensis (Bunge) Regel saponins (PRS) against Oncomelania hupensis (O. hupensis).

METHODSO. hupensis snails were exposed to 40% and 80% of 24 h LC50 of PRS for 24 h, and then choline esterase (CHE), alanine aminotransferase (ALT), alkaline phosphatase (ALP), lactate dehydrogenase (LDH) activities in cephalopodium and liver of snails were determined. Niclosamide (NIC) was used as the reference molluscicide. Zebra fish lethality test was evaluated to non-target aquatic species of PRS.

RESULTSThe molluscicidal activity of PRS (LC50 at 24 h: 0.48 mg/L) was similar to that of NIC (LC50 at 24 h: 0.16 mg/L). Significant alterations about CHE, ALP, and ALT activities both in the cephalopodium and the liver of snails were observed when O. hupensis was exposed to 40% and 80% LC50 of PRS or NIC for 24 h. PRS and NIC could not affect LDH activity in the cephalopodium and the liver. Lower toxicity to fish of PRS was observed up to the highest concentration tested than NIC.

CONCLUSIONPRS, as compared with the reference molluscicide NIC, is thought to be used for the control of harmful vector snails safely.

Animals ; Molluscacides ; pharmacology ; Pulsatilla ; chemistry ; Saponins ; pharmacology ; Snails ; drug effects

To obtain the recombinant pZP3alpha protein for the study of the contraceptive vaccines, the DNA sequence (446-1423) encoding purified pZP3alpha was inserted into a vector--pPICZalphaA. The recombinant plasmid pPICZalphaA-pZP3alpha was linearized and then transformed into Pichia pastoris GS115 by electroporation. Engineering strains were attained by screening with zeocin and induced to produce rpZP3alpha in high-density fermentation. Then rpZP3alpha was purified by Cu2+ metal affinity column chromatography from the separated and concentrated fermentative supernatants. The purified rpZP3alpha was identified by SDS-PAGE and Western blot, and the quantity, purity and rate of recovery of the rpZP3alpha were analyzed by Quantity One software. One male rabbit was immunized with the Cu-NTA-purified rpZP3alpha. The antibody responses against rpZP3alpha and porcine ZP were detected by ELISA and the indirect immunofluorescence. Engineering strains expressing rpZP3alpha in secretion were constructed. A 46kD component named rpZP3alpha which can react with anti-pZP3 antibody was purified from fermentative supernatants of engineering strains and the average yield of purified rpZP3alpha obtained from fermentative supernatants was 8mg/L. The purity and the rate of recovery were up to 92% and 63% respectively. The anti-rpZP3alpha antiserum was prepared by immunization of a male rabbit with purified rpZP3alpha. This anti-rpZP3alpha antiserum could react with rpZP3alpha and purified pZP3 in ELISA and bind to porcine zona pellucida which produced bright green fluorescence in the indirect immunofluorescence. The rpZP3alpha (46kD) protein could be successfully expressed in the Pichia pastoris expression system. And this protein retained the immunogenic activity of natural pZP3.
Animals ; Egg Proteins ; genetics ; metabolism ; Electroporation ; Fermentation ; Immunization ; Male ; Membrane Glycoproteins ; genetics ; metabolism ; Pichia ; genetics ; metabolism ; Rabbits ; Receptors, Cell Surface ; genetics ; metabolism ; Recombinant Proteins ; genetics ; immunology ; secretion ; Swine ; Zona Pellucida Glycoproteins

OBJECTIVE To observe whether human CD4 + T cells could be activated by immuno-globulin D (IgD) via IgD receptor(IgDR)-Lck. METHODS Human CD4+ T cells were purified from peripheral blood mononuclear cells (PBMCs) with microbeads. The viability of T cells were detected by CCK-8. The binding affinity and expression of IgDR on T cells were detected by flow cytometry. The protein expression of IgDR, Lck and P-Lck were analyzed by western blot. RESULTS IgD could concentration-dependent bind to IgDR on CD4+ T cells. The expression of IgDR was increased in response to treatment with IgD in a time- dependent and concentration- dependent manner. Stimulating by IgD resulted in enhanced phosphorylation of Lck compared with that in the medium control sample. The expression of Lck was not changed. As inhibitor of PTK, Herbimycin A or A770041, which combined with IgD could significantly inhibit phosphorylation of Lck(Tyr394). The proliferation promoting effect of IgD was blocked by Herbimycin A or A770041. IgD could stimulate CD4+ T cell activation and proliferation through upreg?ulating activating tyrosine residue of Lck (Tyr394) phosphorylation. CONCLUSION These results demon?strate that IgD exaggerates CD4+T cell activities, which may be through promoting Lck phosphorylation.