BACKGROUND:Caffeic acid phenethyl ester (CAPE) can inhibit lipid peroxidation after rat brain injury. However, the trend of 5-lipoxygenaseis (5-LOX) and cysteinyl leukotrienes (CysLTs) in model of Parkinson’s disease, and whether CAPE protects against rotenone-induced cel ular injuries by inhibiting the levels of 5-LOX and CysLTs stil need further research.
OBJECTIVE:To investigate the protective effect of CAPE on the rotenone-induced Parkinson-like injury, and to determine whether 5-LOX involved.
METHODS:(1) PC12 cel s in good-growth were col ected and divided into five groups cultured with different concentrations of rotenone (0, 0.01, 0.1, 1, 10μmol/L). 24 and 48 hours later, changes of cel ular morphology and activity were observed to single out the optimum concentration of rotenone;at 24 hours, the levels of 5-LOX and CysLTs were detected by western blotting and ELISA, respectively. (2) PC12 cel s were pretreated with different concentrations of CAPE (0, 0.01, 0.1, 1, 10 μmol/L) for 30 minutes, and 1 μmol/L rotenone was then added. The other cel s received no intervention as blank control group. Subsequently, the cel activity was detected, and the CysLTs production was detected by ELISA at 24 hours.
RESULTS AND CONCLUSION:(1) Rotenone (0.1-10μmol/L) could induce PC12 cel injury with overt morphological and cel activity changes at 24 hours, especial y the 1 μmol/L rotenone. (2) Rotenone also significantly increased the 5-LOX expression and CysLTs production in a concentration-dependant manner. (3) CAPE (1-10μmo/L) significantly attenuated rotenone-induced CysLTs production and cel viability reduction in a concentration-dependant manner. (4) These results suggest that CAPE protects against PC12 cel injuries in the model rat with Parkinson’s disease induced by rotenone involving 5-Lox.

The ability to induce hypersensity on leaves of tomato and the stability of double-plasmid of an harpin-producing, nitrogen-fixing engineered strain E4 were tested. Hypelsensitivity-inducing experiment indicated that the time and density of hypersensitivity-induction of E4 was similar to those of DH5, the positive control of pCPP430. Although E4 took the same time to induce hypersensitivity as 308R, another positive control of pCPP430, it induced weaker hypersen- sitivity on tobacco leaves. On tomato leaves, there was no difference in time and density of hypersensitivity between E4 and 308R (pCPP430). Results revealed that the two plasmids, pCPP430 and pMC73A, were unstable in host bacteria, with the losing rate of 100% at the 48th generation. The emergence probability of bacteria with either pCPP430 or pMC73A was almost the same.

AIM: To investigate killing effect of herpes simplex virus thymidine kinase/ganciclovior system(HSV-TK/GCV) on osteosarcoma cell and its mechanisms.METHODS: Recombinant retroviral vector (DORHyTK) containing hygromycin phosphotransferase-thymidine kinase fusion gene(HyTK) was constructed and introduced into human osteosarcoma cell line OS732 with DOTAP. DNA and total RNA extracted from HyTK expressing cells (OS732TK) were tested by PCR and RNA dot blot analysis. A chemosensitivity of OS732TK cells to GCV and “bystander effect” were measured by means of MTT colorimetric assay. Hoeschst 332258 staining and flow cytometric analysis were performed for mechanism of HSV-TK/GCV system gene therapy. RESULTS: DORHyTK was constructed successfully; the HyTK gene existed and expressed in OS732TK cells; All OS732TK cells were killed after 5 days of exposure to 5 mg/L GCV. The “bystander effect” was observed in both a high population and a low population, but the former was stronger than the latter. Hoeschst 332258 staining revealed the characteristic hallmarks of apoptosis, however necrosis also existed. Flow cytometric analysis of DNA content showed a G0-G1 phase blockade. CONCLUSION: HSV-TK/GCV gene therapy system showed its strong killing effects on osteosarcoma cells OS732; The phenomenon of “bystander effect” was also very apparent. GCV exposure induced both necrotic and apoptotic death in HSV-TK expressing cells, and perturbed the cell cycle. HSV-TK/GCV gene therapy system may provide a new therapeutic approach for treatment of osteosarcoma.

OBJECTIVE:To explore the effects of Bushen huoxue huatan decoction on ovarian blood supply and form in model rats with polycystic ovarian syndrome(PCOS). METHODS:50 SD female rats were randomized into normal control group,model group,positive control group [rosiglitazone,3 mg/(kg·d)],Bushen huoxue huatan decoction low-dose and high-dose groups [5.0, 7.5 g/(kg·d)],with 10 rats in each group. Except for normal control group,PCOS model was induced in the other groups by Poresky method. After modeling,treatment groups were given relevant medicine intragastrically. The normal control group and mod-el group were given normal saline [10 mL/(kg·d)] intragastrically,once a day,for 22 days. After the end of administration,the rats were sacrificed. The microvessel density (MVD) was recorded. mRNA expression of vascular endothelial growth factor (VEGF), ovarian angiotensin Ⅱ receptor(Ang Ⅱ)type 1(AT1)and type 2(AT2),G-protein coupled receptor MAS were all detected in ovarian tissue of rats. The ovarian form was observed. RESULTS:Compared with normal control group,MVD of mod-el group and treatment groups were all increased,while mRNA expression of AT1,AT2 and MAS were down-regulated (P<0.05);cystic sinus sollicle,the decrease of granular cell layer,egg mother cells disappearance,the decrease of luteal tissue and other obvious pathological changes could be found in ovarian tissue of rats in model group. Compared with model group,above in-dexes of treatment groups were improved significantly,and the effects of Bushen huoxue huatan decoction low-dose and high-dose groups were better than that of positive control group (P<0.05). The pathological changes of ovarian tissue in treatment groups were all improved to certain extent. CONCLUSIONS:Bushen huoxue huatan decoction can improve ovarian blood supply,angio-genesis and ovulation in PCOS rats. It is beneficial to improve ovarian form.

AIM: To investigate killing effect of herpes simplex virus thymidine kinase/ganciclovior system(HSV-TK/GCV) on osteosarcoma cell and its mechanisms.METHODS: Recombinant retroviral vector (DORHyTK) containing hygromycin phosphotransferase-thymidine kinase fusion gene(HyTK) was constructed and introduced into human osteosarcoma cell line OS732 with DOTAP. DNA and total RNA extracted from HyTK expressing cells (OS732TK) were tested by PCR and RNA dot blot analysis. A chemosensitivity of OS732TK cells to GCV and "bystander effect" were measured by means of MTT colorimetric assay. Hoeschst 332258 staining and flow cytometric analysis were performed for mechanism of HSV-TK/GCV system gene therapy. RESULTS: DORHyTK was constructed successfully; the HyTK gene existed and expressed in OS732TK cells; All OS732TK cells were killed after 5 days of exposure to 5 mg/L GCV. The "bystander effect" was observed in both a high population and a low population, but the former was stronger than the latter. Hoeschst 332258 staining revealed the characteristic hallmarks of apoptosis, however necrosis also existed. Flow cytometric analysis of DNA content showed a G 0-G 1 phase blockade. CONCLUSION: HSV-TK/GCV gene therapy system showed its strong killing effects on osteosarcoma cells OS732; The phenomenon of "bystander effect" was also very apparent. GCV exposure induced both necrotic and apoptotic death in HSV-TK expressing cells, and perturbed the cell cycle. HSV-TK/GCV gene therapy system may provide a new therapeutic approach for treatment of osteosarcoma.

Objective To analyze the association of polymorphisms of estrogen receptor (ER) α and β genes with systemic lupus erythematosus (SLE) in Chinese Han cohort of Yunnan Province.Methods XbaⅠ and Pvu Ⅱ of ERα gene,Rsa Ⅰ and Alu Ⅰ of ERβ gene were typed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) in 697 SLE patients and 638 healthy controls.The frequency distribution of the alleles and genotypes were analyzed by Hardy-Weinberg equilibrium test and x2 test.Results ① For ERα gene,the frequency of minor allele of Pvu Ⅱ C in SLE patients was significantly higher than healthy controls (x2=15.427,P=0.001);the allele frequencies of XbaⅠ in SLE patients showed no significant difference compared with healthy controls (P＞0.05).The frequency of minor genotype of Pvu Ⅱ CC in SLE patients was significantly higher than healthy controls (x2=17.371,P=0.011).The frequency of two locus haplotype AATT in SLE patients was significantly lower than healthy controls (x2=6.333,P=0.012);the frequency of the two locus haplotype AACC in SLE patients was significantly higher than healthy controls (x2=7.771,P=0.038).② For ERβ gene,the frequency of minor allele RsaⅠ A in SLE patients was significantly lower than healthy controls (x2=12.595,P=0.013);the allele frequencies of Alu Ⅰ in SLE patients showed no significant differences compared with the healthy controls (P＞0.05).The frequency of minor genotype AA of Rsa Ⅰ in SLE patients was significantly higher than healthy controls (x2=41.456,P=0.000).The frequency of two locus haplotype AAGG in SLE patients was significantly higher than healthy controls (x2=37.063,P=0.000).The frequency of the two locus haplotype AAGA in SLE patients was significantly lower than healthy controls(x2=21.086,P=0.001).③ Pvu Ⅱ C was related with splenomegaly (x2=4.212,P＜0.05).The two locus haplotype AGTC of Xba Ⅰ and Pvu Ⅱ was related with edema (x2=7.898,P＜0.05).Conclusion There are associations between the polymorphisms of ERα and ERβ genes and SLE.The ERα and ERβ genes may be the susceptible genes for SLE in Yunnan Han Chinese Cohort.

Objective To explore the correlation between hepatitis B virus large surface protein(HBV-LP)and hepatitis B virus deoxyribonucleic acid(HBV-DNA),analyze the clinical significance of detecting the two index dynamically for diagnosis and treatment of hepatitis B.Methods Enzyme linked immunosorbent assay(ELISA)and fluorescent quantitation polymerase chain reaction(FQ-PCR)were used to detect the levels of HBV-LP and HBV-DNA in serum specimen of 230 hepatitis B patients.Results There was a positive correlation between the content of HBV-LP and copy numbers of HBV-DNA in serum of hepatitis B positive cases(r=0.84,P＜0.01).The positive rate was 84.78％ in HBV-LP and 84.35％ in HBV-DNA of 230 HBV positive cases.As a result there was no significant difference(P＞0.05)in this study.Therefore,the positive rate was 63.63 ％(35/55)in HBV-LP and 58.18％(32/55)in HBV-DNA of 55 HBV negative cases.There was also no significant difference(P＞0.05)in this study.However,the positive rate of HBV-LP(82.47％)was higher than HBE(52.06％)in the 194 HBV-DNA positive cases.There was significantly different(P＜0.01).Conclusion HBV-LP and HBV-DNA are the significant index of the degree of hepatitis B virus replication in hepatitis B positive cases,especially in determining the virus replication and prognosis of treatment in HBe Ag negative,providing the reliable laboratory data for the clinical diagnosis and treatment.

Purpose　To observe the killing effect of HSV-TK/GCV system on osteosarcoma cells. Methods　The eukaryotic exprssoin vector (tgCMV-HyTK) containing HSV-TK gene was transduced into human osteosarcoma cell lines SAOS-2 by DOTAP. Total RNA extracted from HSV-TK expressing cells (SAOS-2TK) were tested by RNA dot blot analysis. A chemosensitivity of SAOS-2TK cells to GCV and “bystander effect” were measured by means of MTT colrimetric assay. Results　HSV-TK gene was expressed in SAOS-2TK cells. SAOS-2TK cells were susceptible to the cytotoxicity of GCV. A meaningful “bystander effect” was demonstrated. Conclusion　HSV-TK/GCV system may kill selectively the human osteosarcoma cell line SAOS-2.

Animals ; Diet, High-Fat ; Dietary Fats ; administration & dosage ; Female ; Obesity ; etiology ; physiopathology ; Physical Conditioning, Animal ; physiology ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Receptors, Leptin ; genetics ; metabolism ; Sexual Development ; physiology

Calcium Channel Blockers ; pharmacology ; Cell Survival ; drug effects ; Drug Antagonism ; Humans ; Kidney ; cytology ; drug effects ; Lead ; toxicity