Animals ; Heart Ventricles ; cytology ; Lipid Bilayers ; Potassium Channels ; Sarcolemma ; physiology ; Swine

BACKGROUND: The degree of neurofilament (NF) phosphorylation is closely correlated with the occurrence of Alzheimer disease (AD), and apolipoprotein E4 (apoE4) is a generally acknowledged liability factor for AD, but the effect and mechanism of apoE4 on the NF phosphorylation in neurons are not very clear. It has been reported that in the neurons cultured in vitro and in brain tissue of AD patients, the amino acid residues of apoE4 protein C terminal (272-299) could be truncated by hydrolysis,and produce truncated-apoE4 fragment. The latter interacts with the NF phosphorylation in neurofibrillary tangles (NFTs), which are the characteristic pathological changes of AD.OBJECTIVE: To observe the effect of truncated-apoE4 overexpression on the NF phosphorylation in the cultured neurons.DESIGN: A non-randomized controlled observation.SETTING: Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Tongji Medical College, Huazhong University of Science and Technology.MATERIALS: The experiment was carried out in the Laboratory of Biochemistry and Molecular Biology, Tongji Medical College, Huazhong University of Science and Technology in December 2005. The mice neuroma cell strain N2a was provided by Dr. Xu.METHODS: The truncated-apoE4(△272-299) cDNA was subcloned into pEGFP-c3 to form pEGFP-T-apoE4 recombinant. Then pEGFP-c3, pEGFP-apoE4 and pEGFP-T-apoE4 were transfected into N2a cells by lipofectamine2000 respectively. NF phosphorylation was detected by Western blot assay. The activities of glycogen synthase kinase-3 (GSK-3) and cyclin-dependent kinase 5(CDK-5) were measured.MAIN OUTCOME MEASURES: The degree of NF phosphorylation and the activities of GSK-3 and CDK-5 were mainly observed.RESULTS: In the transfected groups, the contents of phosphorylated NF were significantly increased, the GSK-3 activities were significantly increased, which were the most significant in the pEGFP-T-apoE4 transfected group (P＜0.05), but the CDK-5 activities were not significantly different from that in th e control group (P＞0.05).CONCLUSION: These results indicate that in vitro overexpression of truncated-apoE4(△272-299) can lead to NF hyperphosphorylation by activating of GSK-3 but not CDK-5, which may be the underline mechanism of AD induced by truncated-apoE4(△272-299).

Objective To investigate the influences of mutation at precore and basic core promoter(BCP) region in hepatitis B virus(HBV) on the immune response of specific cytotoxic T lymphocytes(CTL) in patients with chronic hepatitis B(CHB).Methods The number of specific CTL in peripheral blood mononuclear(PBMC) of CHB patients were tested by cytokine flow cytome- try(CFC) and HBV core18-27 peptide.HBV precore and BCP fragments were directly sequenced. Results Twenty-one(38.9%) samples were HBV precore G1896A mutation.Twenty-six(48.1%) samples were BCP region 1762/1764 combined mutation.Thirteen(24.1%) stains were three sites mutated simultaneously.Stimulated with HBV core 18-27 in vitro,the specific CTL level was signifi- cantly higher in the patients with G1896A mutation and BCP region mutation [(0.41?0.09)%, (0.36?0.08)%,(0.48?0.08)%,respectively]than those without mutation[(0.11?0.06)%, P

Cathepsin B from Helicoverpa armigera (HCB) belongs to the group of cysteine proteinases. HCB is proved being involved in the degradation of yolk proteins during embryonic development,which is an acidic preferring enzyme and is resistant to SDS. The expression of the proenzyme may offer a model for investigating the activation of the enzyme. The HCB gene was constructed into pPIC9K and expressed in Pichia pastoris KM71 strain . After induction by methanol, HCB was expressed and secreted into the medium. The molecular weight of the recombinant procathepsin B was determined as about 38 kDa. The expressed product was confirmed to be HCB by immunoblotting assay using specific rabbit anti-HCB polyclonal antibody. The activity of the product was assayed by in situ hydrolysis (gelatin-SDS-PAGE). These results showed that HCB with proteolytic activity was expressed in P. pastoris KM71. This proenzyme can be used for further research on the activation of the proenzyme or industrial production.

Objective To study the increased radiosensitivity of tumor cells by triterpene acids of loquat leaf (TAL) and mechanism.Methods C6 and MG63 cells were pretreated by TAL,and then were exposed to X-rays at 1,2,3,5,7 Gy.Clonogenic assay was used to evaluate those tumor cells survival fraction (SF) to calculate the sensitization enhancement ratios (SER) of TAL.The cellular micronuclei ratios of tumor cells were analyzed by micronuclei assay.Additionally,the changes of Rad51 and XRCC4 levels were observed by RT-PCR and Western blot.Results D0 values were decreased to 1.31 and 2.85 Gy in C6 and MG63 cells by TAL pretreatment respectively.The SER value of the effect of TAL on C6 and MG63 cells was 1.73 and 2.04,respectively.There was a statistical difference in the cellular micronuclei ratios between tumor cells with TAL above 2 Gy and those without TAL (C6:t =-8.372--2.476,P ＜0.05 ; MG63:t =-4.03--2.557,P ＜ 0.05).TAL attenuated the expression of XRCC4 at transcriptional and translational level,but not for Rad51,a key gene in homologous recombination repair (HRR).Conclusions TAL pretreatment could increase the lethal effect of X-rays on tumor cells in vitro.The mechanism might be involved in the inhibition of non-homologous end joining (NHEJ).

Animals ; Bone Morphogenetic Protein 2 ; metabolism ; Bone Morphogenetic Protein 4 ; metabolism ; Carrier Proteins ; pharmacology ; Liver ; metabolism ; Liver Regeneration ; Male ; Rats ; Rats, Wistar

Through collecting and collating the development process of traditional Chinese medicine dispensing, the development of modern Chinese medicine dispensing on the basis of experience could be promoted. "Heyaofenji", "Hehe", " Heji" in ancient Chinese medicine, herbal medicine literature and law were collected, and then things were sorted out according to traditional Chinese medicine dispensing theory, skills and legal norms. Firstly, "Tang Ye Jing Fa" is the earliest book which marks the rudiment of traditional Chinese medicine dispensing. Secondly, traditional Chinese medicine dispensing theory formed in "Shen Nong's herbal classic". Thirdly, Zhang Zhongjing's "Treatise on Febrile Diseases" marked the formation of Chinese medicine dispensing skills. Lastly, Provisions in Tang Dynasty law marks the development of traditional Chinese medicine dispensing.
Chemistry, Pharmaceutical ; education ; history ; legislation & jurisprudence ; methods ; China ; Dosage Forms ; Drug Compounding ; history ; methods ; Drug and Narcotic Control ; history ; legislation & jurisprudence ; Drugs, Chinese Herbal ; chemistry ; History, Ancient ; Medicine in Literature ; Medicine, Chinese Traditional ; history ; methods

OBJECTIVETo explore the laws of anovulatory infertility patients of Gan-yin deficiency syndrome (GYDS), and to analyze the correlation between GYDS and partial sex hormones and metabolic parameters.

METHODSRecruited were 103 anovulatory infertility patients, including 48 of GYDS and 55 of non-GYDS. At the same time, 20 healthy pluripara at the child-bearing period were recruited as the control group. The body mass index (BMI) and waist to hip ratio (WHR) were measured. Serum levels of luteinizing hormone (LH), follicle stimulating hormone (FSH), testosterone (T), prolactin (PRL), estradiol (E2), sex hormone binding globulin (SHBG), fasting blood glucose (FBG), fasting insulin (FINS), total cholesterol (TC), triglyceride (TG), low density lipoprotein-cholesterol (LDL-C), high density lipoprotein-cholesterol (HDL-C), and leptin were detected. The inter-group difference of the above indices was compared between the two groups.

RESULTSLevels of BMI, FINS, HOMA-IRI, leptin, TC, TG, LDL-C, T, PRL, and LH were higher in the GYDS group and the non-GDS group than in the control group (P < 0.01, P < 0.05), while SHBG was lower in the GYDS group and the non-GYDS group than in the control group (P < 0.01). Only the PRL level was higher in the GYDS group than in the non-GYDS group (P < 0.01).

CONCLUSIONSGan-yin deficiency is a predominant manifestation in anovulatory infertility patients. Partial disorder of some sex hormones and metabolic derangement might be common pathological factors for anovulatory infertility, while increased PRL levels was dominant in GYDS.

Adult ; Anovulation ; metabolism ; Body Mass Index ; Case-Control Studies ; Female ; Gonadal Steroid Hormones ; blood ; Humans ; Infertility, Female ; metabolism ; Lipids ; blood ; Prolactin ; blood ; Yin Deficiency

Humans ; Minimally Invasive Surgical Procedures ; utilization

To study the ecological distribution and diversity of endophytic fungi associated with Ferula of medicinal plants in Xinjiang. The endophytic fungi were isolated from roots, stems and leaves of Ferula by microbiology research methods and technology. The endophytic fungi were identified using ITS rDNA sequence analysis and morphology analysis. The composition, diversity and preference of endophytic fungal community were analyzed with Shannon-Wiener biodiversity index (H') and Sorensen coefficient (Cs). A total of 337 strains endophytic fungi were isolated and classified into 38 genera, Alternaria, Aureobasidium and Fusarium were the dominant genera. Among the 337 isolates, the endophytic fungi of F. sinkiangensis were the most, The Shannon-Wiener biodiversity index (H') associated with roots of F. fukanensis was the highest, reached 1.85. The highest Sorensen coefficient ( Cs) was between leaf of F. sinkiangensis and leaf of F. ovina, reached 0.75. From the result, endophytic fungi were widely distributed in six Ferula, there are some notable differences between distribution and composition of the endophytic fungi isolated from different issues and different species of Ferula, show a certain degree of species and tissue preference. The results obtained in this study will provide realistic basis and theoretical basis for further study the secondary metabolites of endophytic fungi associated with Ferula, and the relationship between endophytic fungi and their host plants.
Biodiversity ; Ecology ; Endophytes ; isolation & purification ; Ferula ; microbiology ; Fungi ; classification ; isolation & purification ; metabolism