OBJECTIVETo observe monocyte (Mo) development in wild type C57BL/6 mice and apoE gene knockout (apoE(-/-)) mice, and to evaluate the immuno-regulatory effect of Huanglian Jiedu Decoction (HJD) on peripheral Mo development in apoE(-/-) mice.

METHODSFour, 8, 12, and 16 weeks old female C57BL/6 mice were set up as control groups of different ages, while 4, 8, 12, and 16 weeks old female apoE(-/-) mice were set up as hyperlipidemia groups of different ages. Four-week old female C57BL/6 mice were recruited as a blank group. Four-week old female apoE(-/-) mice were randomly divided into the control group, the Western medicine group, and the Chinese medicine group by paired comparison, 5 in each group. Equivalent clinical dose was administered to mice according to body weight. Mice in the Western medicine group were administered with Atrovastatin at the daily dose of 10 mg/kg by gastrogavage, while those in the Chinese medicine group were administered with HJD at the daily dose of 5 g/kg by gastrogavage. Body weight was detected each week. After 4 weeks blood lipids levels (such as TG, TC, LDL-C, and HDL-C), and the proportions of Mo and Ly6c(hi) were detected.

RESULTSCompared with 4-week-old homogenic mice, the proportion of Mo decreased in 16-week-old C57BL/6 mice (P < 0.05). Levels of TC and TG, and the proportion of Ly6c(hi) subtype increased, but the proportion of Mo de- creased in 8-week-old apoE(-/-) mice (P <0. 05). Levels of TC, TG, and LDL-C increased in 12-week-old apoE(-/-) mice (P < 0.05). Levels of TC, TG, LDL-C, and HDL-C increased in 16-week-old apoE(-/-) mice (P < 0.05, P < 0.01). Compared with 8-week-old homogenic mice, the proportion of Mo decreased in 16-week-old C57BL/6 mice (P < 0.05); levels of TC and LDL-C increased in 12-week-old apoE(-/-) mice (P < 0.05); levels of TC and HDL-C increased in 16-week-old apoE(-/-) mice (P < 0.05, P < 0.01). Compared with C57BL/6 mice of the same age, TC and TG increased, HDL-C decreased (P < 0.01) in 4-and 8-week-old apoE(-/-) mice (P < 0.01); levels of TC, TG, LDL-C increased, and HDL-C level decreased in 12- and 16-week-old apoE(-/-) mice (P < 0.05, P < 0.01); the proportion of Mo increased in 4-week-old apoE(-/-) mice (P < 0.05); proportions of Mo and Ly6c(hi) increased in 8-week-old apoE(-/-) mice (P < 0.05). Compared with the blank control group, levels of TC, TG, and LDL-C, proportions of Mo and Ly6c(hi) increased (P < 0.01, P < 0.05), but HDL-C level decreased (P <0. 01) in the control group after intervention. Compared with the control group, body weight gained less in the Western medicine group and the Chinese medicine group (P < 0.05); the proportion of Ly6c(hi) subtype decreased in the Chinese medicine group (P < 0.05).

CONCLUSIONSIn development process blood lipids levels in apoE(-/-) mice are not only associated with age. Blood lipids levels induced growth changes in natural immune system are also correlated with age. In early stage of lipids development HJD intervention could correct this special immune disorder in apoE(-/-) mice.

Animals ; Apolipoproteins E ; genetics ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; Gene Knockout Techniques ; Hyperlipidemias ; Lipids ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; Monocytes ; physiology

The study is to establish an HPLC method using fluorescence detector for the determination of doxazosin enantiomers and investigate their chiral inversion in vitro and in vivo. Ultron ES-OVM was taken as the chiral chromatographic column, and the column temperature was 30 degrees C. Isocratic elution using a mobile phase of phosphate buffer-acetonitrile (85 : 15, v/v) at a flow rate of 0.8 mL x min(-1) was done. The fluorescence detection was set at lambda(Ex) = 255 nm and lambda(Em) = 385 nm. Prazosin was used as the internal standard. (-) Doxazosin or (+) doxazosin added into rat plasma in vitro was determined after incubating in 37 degrees C water bath for 2, 5 and 10 days. (-) Doxazosin or (+) doxazosin was administered orally to the rats for one months. Plasma samples were taken at 8 h after the last administration. A good linear relationship was achieved when the concentration of doxazosin enantiomers was within the range of 4 - 2 000 ng x mL(-1). The average recovery for (-) doxazosin was 99.5% with RSD 3.6%, and for (+) doxazosin was 99.3% with RSD 4.3%. Chiral inversion was observed neither in vitro nor in vivo studies. The method is selective, accurate and reproducible, which is suitable for the detection of doxazosin enantiomers in rat plasma. The in vitro and in vivo studies indicate that chiral inversion occurs uneasily between (-) doxazosin and (+) doxazosin in the rat.
Animals ; Blood Chemical Analysis ; methods ; Doxazosin ; blood ; chemistry ; Male ; Rats ; Rats, Sprague-Dawley ; Reproducibility of Results ; Sensitivity and Specificity ; Stereoisomerism

OBJECTIVETo investigate the protective effect of suppressive oligodeoxynucleotides (Sup ODN) on interferon-γ (IFN-γ) and signal transducers and activators of transcription (pSTAT4) expression of Silica-induced pulmonary inflammation in Mice.

METHODSSixty Balb/c mice were randomly divided into 4 groups, normal control group, silicious group, suppressive oligodeoxynucleotides (Sup ODN) group, control oligodeoxynucleotides (Con ODN) group. Except the normal control group injected normal saline, the rest groups were induced by the intratracheal instillation of 0.1 ml (5 g/L) of sterilized silica suspension. Sup ODN group and Con ODN group were treated by i.p. injection of 0.3 ml (1mg/mL) of suppressive or control ODN 3 h before silica administration. After 7 days, the animals were killed and levels of IFN-γ were detected by ELISA. The pathologic changes in lung tissues of mice were observed with HE staining. Expressions of IFN-γ and pSTAT4 in lung tissue were detected with immunohistochemistry and quantified by Image-Pro Plus 7.0.

RESULTSHE staining showed that the lung tissue of silicious group were damaged seriously than Sup ODN group. Compared with the normal control group (serum: (280.1±41.3) pg/ml, lung tissue: (0.249±0.373), IFN-γ increased in silicious group (serum: (886.3±81.7) pg/ml, lung tissue: (0.270±0.300) (P < 0.05). Compared with the normal control group and Con ODN group [(894.5±91.6) pg/ml], IFN-γ in the serum of Sup ODN group decreased significantly (P < 0.01). Compared with the silicious group , IFN-γ in lung tissue decreased in Sup ODN group (0.241±0.250) (P < 0.05). Compared with the normal control group (0.279±0.353), pSTAT4 in lung tissue increased significantly in silicious group (0.313±0.231) (P < 0.01). Compared with the silicious group, pSTAT4 in lung tissue decreased significantly in Sup ODN group (0.269±0.523) (P < 0.01).

CONCLUSIONSup ODN attained protective effect on Silica treated mice by suppressing expression of IFN-γ and pSTAT4.

Animals ; Female ; Inflammation ; metabolism ; Interferon-gamma ; metabolism ; Lung ; drug effects ; metabolism ; pathology ; Mice ; Mice, Inbred BALB C ; Oligodeoxyribonucleotides ; pharmacology ; Phosphorylation ; STAT4 Transcription Factor ; metabolism ; Silicon Dioxide ; toxicity

In recent years, iron has been regarded as a common pathological feature of many neurodegenerative diseases, including Alzheimer's disease (AD), Parkinson's disease (PD) and Friedreich's ataxia (FRDA). A number of genes involved in iron transport, storage and regulation have been found associated with initiation and progression of neurodegeneration. However, whether iron abnormalities represent a primary or secondary event still remains unknown. Due to the limitation in transgenic rodent model construction and transfection systems, the progress in unraveling the pathogenic role of different iron-related proteins in neurodegenerative diseases has been slow. Drosophila melanogaster, a simple organism which has a shorter lifespan and smaller genome with many conserved genes, and captures many features of human nervous system and neurodegeneration, may help speed up the progress. The characteristics that spatial- and temporal-specific transgenic Drosophila can be easily constructed and raised in large quantity with phenotype easily determined turn Drosophila into an excellent in vivo genetic system for screening iron-related modifiers in different neurodegenerative conditions and hence provide a better picture about the pathogenic contribution of different iron-related protein abnormalities. It is believed that identification of important iron-related genes that can largely stop or even reverse degenerative process in Drosophila models may lead to development of novel therapeutic strategies against neurodegenerative diseases.
Alzheimer Disease ; physiopathology ; Animals ; Disease Models, Animal ; Drosophila melanogaster ; Friedreich Ataxia ; physiopathology ; Humans ; Iron ; Neurodegenerative Diseases ; physiopathology ; Parkinson Disease ; physiopathology

OBJECTIVETo investigate the possible association between ZNF230 gene and azoospermia.

METHODSScreening for mutation of all 6 exons of ZNF230 gene was performed by denaturing high performance liquid chromatography(DHPLC) in 99 patients with azoospermia and in 115 healthy men as controls.

RESULTSAn A-->G transition at nucleotide 316 in exon 6 was identified. There were significant differences in the distribution profiles of both allele and genotype frequencies between patient group and control group (P < 0.01 and P < 0.05, respectively). In addition,there was a statistically significant difference in the serum follicle stimulating hormone (FSH) level between the patients with GG/GA genotype and those with AA genotype (P < 0.05).

CONCLUSIONZNF230 gene may be associated with azoospermia, and the A316G mutation may be correlated with the serum FSH level.

Adult ; Azoospermia ; diagnosis ; genetics ; Base Sequence ; Chromatography, High Pressure Liquid ; DNA Mutational Analysis ; DNA-Binding Proteins ; genetics ; Gene Frequency ; Genetic Testing ; Genotype ; Humans ; Male ; Mutation ; Polymerase Chain Reaction ; Transcription Factors ; genetics ; Young Adult

The flagellin of Listeria monocytogenes is encoded by flaA gene;there is no detail report about the influence of flaA gene on the virulence of LM90 SB2.FlaA gene-deleted strain was constructed successfully with recombination technology.The influence of FlaA on LM90 SB2 virulence was evaluated by motility,BF formation,LD50,etc.Results showed that the colony morphology did not change,gene-deleted strain still had good genetic stability,but its growth was slow and the motility was decreased in the environment at 25 ℃,the morphological structure of the mutant BF was looser and incomplete,LD50 was increased from 4.27 × 106 cfu to 1.62 × 107 cfu.Results indicated that the flaA gene affected the flagellar formation of LM90 SB2 significantly,the ability of the BF formation and the virulence of the flaA deleted strain were decreased obviously.

OBJECTIVE To study the effects of Liuweidihuang Formula (LWDHF) on the ovariectomized ApoE-/-AS mice and the proliferation and migration of vascular smooth muscle cell (VSMCs) in vitro,and to elucidate the roles of estrogen receptor β (ERβ) and heat shock protein 27 (HSP27) phosphorylation in the context.METHODS The artery injury of ApoE /-mice was observed by HE staining and the phosphorylation of HSP27 in vessels was detected by immunohistochemistry.VSMC proliferation and cell cycle were examined by MTT assay and flow cytometry,respectively.VSMC migration was observed by wound-healing assay and F-actin staining.The levels of HSP27,pHSP27,cyclin D1,protein phosphatase 2 (PP2A) and ERβ were determined by Western blot analyses.Transfection with ERβ siRNA and ERα shRNA was carried out to investigate their roles in LWDHF effects,respectively.RESULTS LWDHF improved the arterial lesion,inhibited HSP27 phosphorylation,and increased the expression of PP2A and ERβ in ovariectomized ApoE / AS mice.In in vitro experiments,angiotensin Ⅱ (Ang Ⅱ) at 1 × 10-7 mol/L obviously induced VSMC proliferation and migration,increased the expression of cyclin D1 and cell number in the S phase,but these effects were significantly inhibited by LWDHF (12μg/mL).Further molecular examinations showed that LWDHF up-regulated PP2A expression through ERβ to inhibit HSP27 phosphorylation,contrib uting to inhibition of VSMC proliferation and migration.CONCLUSION LWDHF reduced menopause AS in vivo and inhibited proliferation and migration of VSMCs by regulating ERβ-inhibition of HSP27 phosphorylation in vitro.These discoveries provided novel molecular insights into LWDHF as potential therapy for AS.

It is the focus of higher education reform in the new era to comprehensively promote the con-struction of ideology and political education based on the characteristics of professional courses and en-hancing the effectiveness of ideology and political education.As an important basic course for medical students in colleges,molecular biology is closely related to basic medical disciplines and clinical medi-cine,and is a rapidly developing cutting-edge discipline,which has the natural advantage of serving as a carrier of ideology and political education.In this study,the innovative integration of project-based group study (PBGS) with the outcome-oriented behavior (OBE) of moral education is applied to the teaching of the ideology and politics of the medical molecular biology course,and the integration of the two has made a useful exploration to enhance the effectiveness of the ideology and politics teaching of the course.Taking students as the center,we have constructed an ideology and politics teaching system for medical molecular biology courses by combining on-line and off-line teaching activities through improving the teaching objectives,innovating the teaching design,digging into the case of ideology and politics,intro-ducing a variety of teaching methods,strengthening the management of teaching practice,and optimizing the evaluation mode.After two years of teaching practice,this model has effectively improved the teach-ing effect of the medical molecular biology course.The academic performance of the students in the prac-tice group has improved significantly,and the teachers and students have been given excellent evalua-tion.The results of the questionnaires before and after class showed that more than 80％ of the students believed that their horizons had been broadened and their knowledge had been increased through learn-ing.More than 50％ of the students believed that their learning ability and innovation consciousness had been improved;their scientific research quality had been improved;and their confidence in studying medicine had been strengthened.By strengthening the cultivation of students' scientific research and in-novation capabilities,we guided students to participate in subject competitions and won many national a-wards.Throughout the teaching process,we aim to expand the breadth and depth of ideology and political education,cultivate scientific spirits,innovation ability,moral cultivation,and humanistic qualities.In sum,our work provides experiences for the cultivation of high-quality medical talents.

Objective: The current study aims to explore precipitating and social risk factors for internet addiction (IA) in university undergraduate students, and to provide evidence for interventions and the early prevention of IA in different genders. Methods: Four thousand eight hundred and fifty-eight college sophomores completed an online survey on their internet use-related behaviours and social risk factors. Results: We found that more male (8.3%) than female students (5.4%) had moderate and severe IA. The main online activity in the moderate and severe IA groups was online gaming in males and online streaming in females. Roommates engaging in similar internetbased entertainment was a risk factor of IA only for males, while not being in a romantic relationship was a risk factor of IA for females only. Infatuation with the internet before college and adjustment problems for college life were shared risk factors for both genders in the mild and moderate IA groups. Conclusion IA was a common phenomenon in college students with shared and unique precipitating and social risk factors in males and females. The gender-sensitive risk factors for IA warranted earlier and individualized intervention and prevention strategies for IA in this population.

Objective: The current study aims to explore precipitating and social risk factors for internet addiction (IA) in university undergraduate students, and to provide evidence for interventions and the early prevention of IA in different genders. Methods: Four thousand eight hundred and fifty-eight college sophomores completed an online survey on their internet use-related behaviours and social risk factors. Results: We found that more male (8.3%) than female students (5.4%) had moderate and severe IA. The main online activity in the moderate and severe IA groups was online gaming in males and online streaming in females. Roommates engaging in similar internetbased entertainment was a risk factor of IA only for males, while not being in a romantic relationship was a risk factor of IA for females only. Infatuation with the internet before college and adjustment problems for college life were shared risk factors for both genders in the mild and moderate IA groups. Conclusion IA was a common phenomenon in college students with shared and unique precipitating and social risk factors in males and females. The gender-sensitive risk factors for IA warranted earlier and individualized intervention and prevention strategies for IA in this population.