Objective To screen out the PI3K/Akt pathway related genes in vascular smooth muscle cells (VSMCs) subjected to cyclic strain. Method VSMCs of rat aorta were subjected to cyclic strain (10 %, 1 Hz) by using a FX-4000T system. Phosphorylation of Akt was examined by Western blotting. Suppression subtractive hybridization (SSH) method was employed to analyze the differently expressed cDNA sequence between the strained VSMCs pretreated with and without wortmannin, a specific inhibitor of PI3K. These fragments were ligated with T vectors, screened through the blue-white screening system to establish cDNA library, and was sequenced and analyzed in GenBank with BLASTN search.Result Level of Akt phosphorylation of VSMCs was significantly enhanced by the mechanical strain compared with the control. Ten different expressed sequence tags (EST) were gained after sequencing 30 clones randomly selected from 54 white clones. There may be six genes related with the mechanical strain and cell signal PI3K/Akt, such as High mobility group box 1 (HMGB1), Neural precursor cells expressed (Nedd4a), Cyclin-dependent kinase 5 (CDK5), Histone deacetylase 3 (HDAC3), Ubiquitin-like 1 (Uble1a) and Heterogeneous nuclear ribonucleoprotein H1 (hnRNP). Conclusion SSH is an effective and reliable approach to screen out the genes related with the mechanical strain and PI3K/Akt pathway in VSMCs.

Objective To build experimental animal model of cerebral alveolar echinococcosis disease of sheep,in order to study of human alveolar hydatid disease of the brain.Methods Experiment animal models of ten Xinjiang big-tail sheep were performed by, direct skull puncture,intracerebral inoculation of echinococcus multilocularis.MRI was used to observe the growth status of cerebral alveolar echinococcosis disease of sheep after 8 months,and morphological and pathological characteristics after autopsy were ana-lysed.Results 4 sheep models (40%)were successful built which were confirmed by pathology and MRI.On MRI,4 cases all were single lesion,on T2 WI there was multiple follicles bubbles under the background of low signal in one case,and low signal in other three cases.Under the microscope,a large number of lymphocytes,eosinophils and plasma cells infiltrated the lesion area,around which small blood vessels were blocked and had inflammatory reaction were showed.Conclusion The method using artificial inocula-tion rat alveolar echinococcosis to establish experimental model of cerebral alveolar echinococcosis disease of sheep has the character-istic of feasibility,simplicity and repeatability.

Objective To determine in vitro drug susceptibility to five antifungal agents of clinical Cryptococcus neoformans strains isolated from different areas of China in recent ten years. Methods Eighty clinical isolates of Cryptococcus neoformans were isolated from Shanghai, Guangdong, Fujian, Beijing and some other areas of China from 1998 to 2007. The minimal inhibitory concentrations (MIC) of the isolates to five antifungal agents, including amphotericin B, fluconazole, flucytosine, itraconazole and voriconazole, were determined using broth microdilution procedure (document M27-A2) recommended by the Clinical and Laboratory Standards Institute (CLSI). Kruskal-Wallis rank sum test was employed for the statistical analysis. Results The MIC50 of the Cryptococcus neoforrnans isolates tested for amphotericin B, fluconazole, flucytosine, itraconazole and voriconazole were 0.5, 4, 2, 0.25 and ≤0.031 3 mg/L, respectively; and the MIC<,90> of the isolates tested for the above antifungal agents were 1, 8, 4, 0.5 and 0.062 5 mg/L, respectively. Among the tested isolates, 3 (3.8 %) were resistant to flucytosine, 4 (5.0 %) were resistant to itraconazole. All isolates were susceptible to amphotericin B and voriconazole. There was no significant difference in MIC of the strains isolated from any particular years to the five agents (χ2=0.500,2.687,2.211, 2.660,0.677,P>0.05). Conclusions The Cryptococcusneoformans isolates are highly susceptible to the five antifungal agents, while a few strains are resistant to flucytosine or itraconazole. The drug susceptibilities of the strains isolated from particular years are similar.

To analyze the relationship between obesity with related measurable indicators and insulin resistance among the residents over 50-year-old in Wuxi area.The questionnaire survey,physical examination,and relevant biochemical measurements as well as gender,age,height,body w eight,waist circumference,and hip circumference were obtained.The prevalence of obesity was 11.95％ of which 57.24％ were with central obesity,and the prevalence in male was higher than that in female(P＜0.05).With inereasing body mass index (BMI) and waist circumference(WC),the prevalence of insulin resistance increased gradually (P ＜ 0.05).Meanwhile,With the increasing HOMA-IR,the prevalence of obesity and central obesity also gradually increased (P＜0.05).BMI,WC,and waist-to-hip ratio can effectively predict insulin resistance.The cut-off point of waist circumference was 87.5 cm for male,and 84 cm for female.At the same level of body mass index,metabolic measurements in central obesity group were higher than those of non-central obesity group with the same body mass index,especially in normal weight group.The related metabolic measurements of metabolic obesity but normal weight group were mostly higher than metabolically healthy but obese group,and there were statistically significant differences in fasting blood glucose,fasting insulin,HOMA-IR and HOMA-β.Obesity,especially central obesity,is closely related to insulin resistance among residents over 50-year-old in Wuxi area.Meanwhile,waist circumference may effectively predict insulin resistance and may serve as a parameter in the occurrence of metabolic syndrome and cardiovascular diseases.

Objective To explore the clinical signiifcance of combined detection of thyroid stimulating hormone (TSH) and free thyroxin (FT4) in dried blood spots in screening for congenital hypothyroidism (CH) in neonates. Methods The TSH and FT4 levels in dried blood spot were measured by time-resolved lfuorescence immunity in live born neonates from June to December 2013. If the screening was positive, the blood was drawn and the serum TSH and FT4 were measured and compared with the results from dried blood spots. Results In a total of 31 199 neonates screened, 12 cases were diagnosed with CH and the prevalence rate of CH was 1/2 600;4 cases were hyperthyropinemia and no pituitary CH was detected. There was no signiifcant difference between TSH or FT4 levels in dried blood spot and those in serum in neonates diagnosed with CH (P>0.05). Conclusions Combined detection of TSH and FT4 in dried blood spot can be used for neonatal screening of CH. It can be applied for early distinguishing CH from hyperthyropinemia, and also helpful for early diagnosis and treatment of central CH.

Objective To explore the regulation mechanism for miR - 125a - 5P in epidermal growth factor receptor(EGFR)signaling pathway in medulloblastoma. Methods The potential targets of miR - 125a - 5P in the EGFR signaling pathway were predicted by TargetScan and Sanger software,there were 3 groups:control group,non -sense group and miR - 125a - 5P group. Their relationship,between miR - 125a - 5P and cyclin - dependent kinase in-hibitor 2B( CDKN2B),E2F transcription factor 3( E2F3),mitogen - activated protein kinase 14( MAPK14)and growth factor receptor - bound protein 10(GRB10),were tested by luciferase experiments. After miR - 125a - 5P oligo-nucleotide was transfected to D341 cells,miR - 125a - 5P level was detected by reverse transcription polymerase chain reaction. Then the thiazolyl blue tetrazolium bromide assay was used to draw the cell growth curves,and Transwell assay was used to detect cell migration ability. The expression levels of GRB10,EGFR,phosphatidylinositol 3 - kinase(PI3K) and Ras were tested by Western blot method. Results The results of luciferase experimental results showed that GRB10 was the only target gene of miR - 125a - 5P. After miR - 125a - 5P being transfected,the D341 cell prolifera-tion obviously declined markedly. Compared with control group[(38. 16 ± 7. 47)% ]and the non - sense group [(36. 79 ± 8. 94)% ],cell migration rate in the miR - 125a - 5P group was lowest[(13. 59 ± 4. 41)% ],and there was a significant difference among 3 groups(χ2 = 11. 495,P < 0. 05);in the miR - 125a - 5P group,the expression level of EGFR increased 1. 67 times,GRB10,PI3K and Ras levels were reduced to 23% ,61% and 42% . Conclusion miR - 125a - 5P can inhibit tumor growth by silenced GRB10 expression targeting EGFR downstream signaling pathways in medulloblastoma.

OBJECTIVE:To systematically review the efficacy of agomelatine in the treatment of depression in acute phase, and provide evidence-based reference for the clinical treatment. METHODS:Retrieved from Cochrane Library,Medline,EMBase, CJFD,VIP,CBM,Wanfang Database,WHO Clinical Trials Registry Platform and American Clinical Trials Registry Platform, the randomized controlled trials (RCT) about agomelatine versus placebo in the depression patients in acute phase. After quality evaluation and data extraction,Meta-analysis was conducted by using Rev Man 5.2 statistics software. RESULTS:A total of 7 RCT were included,involving 2 378 patients. Results of Meta-analysis showed the effective rate in agomelatine group was obviously bet-ter than placebo group [RR=1.43 ,95%CI(1.29 ,1.59),P<0.001] ,remission rate was obviously better than placebo group [RR=1.27 ,95%CI(1.03 ,1.57),P=0.02] ,and the endpoint score of depression scales was obviously lower than placebo group [MD=-2.92,95%CI(-3.65,-2.20),P<0.001],there was statistically significance. CONCLUSIONS:Agomelatine is effective in the treatment of depression patients in acute phase. However,due to the limit of methodological quality and sample size,it remains to be further verified with more rigorously designed and long-term follow-up of large-scale RCT.

Forensic entomotoxicology is a branch of forensic medicine, which applies entomology, toxicology and other related studies to solve the poisoning cases. It has an obvious advantage in the investigation on poisoning death. Based on the expounding definition and research of entomotoxicology, this paper reviews research progress and application value in some aspects of forensic medicine, such as the effects of drugs/toxins on the growth and development of sarcosaphagous insects and the qualitative and quantitative analysis of the drugs/toxins in the poisoned body tissue.
Animals ; Death ; Entomology/methods* ; Forensic Medicine/methods* ; Humans ; Insecta ; Postmortem Changes

OBJECTIVETo identify the expression characteristics of the 1700001022RIK (RIKEN cDNA 1700001022) gene in mice and explore its function by bioinformatic analysis.

METHODSUsing the expression profile of gene microarray, we detected the expression of a new testis-specific gene, 1700001022RIK, in mice. We analyzed its expression characteristics in the testis tissue and their changes in different developmental stages of the testis by RT-PCR, real-time RT-PCR, Western blot, and immunohistochemistry. We performed bioinformatic analysis using a bioinformatic software.

RESULTSThe 1700001022RIK gene was specifically expressed in the mouse testis in an age-dependent manner, most highly in the adult mice. The 1700001022RIK protein was mainly expressed in the spermatogonia, spermatocytes, and round spermatids of the adult mice. Bioinformatic analysis showed that the 1700001022RIK protein amino acid sequence had a high similarity in human and mice, which indicated that this gene was highly conserved in mammals.

CONCLUSION1700001022RIK is a testis-specific gene mainly expressed in the spermatogonia, spermatocytes, and round spermatids of seminiferous tubules, which might be involved in the regulation of spermatogenesis.

Age Factors ; Animals ; Blotting, Western ; Computational Biology ; DNA, Complementary ; Gene Expression ; Genomics ; Male ; Mice ; Molecular Chaperones ; genetics ; Seminiferous Tubules ; Spermatids ; Spermatocytes ; Spermatogenesis ; genetics ; Spermatogonia ; Testis

AIMTo investigate the role of motilin in paraventricular nucleus (PVN) of hypothalamus on the regulation of gastric motility and the mechanism.

METHODSImmunohistochemistry and microinjection of motilin into PVN were used to observe motilin neural cells in PVN, the neural path between PVN and dorsal vagal complex (DVC) and the changes of gastric motility in conscious rats.

RESULTS(1) There were motilin immunoreactive-cells in PVN and a significant increase was found in groups of fasting and HCL-perfusion into duodenum. (2) HRP-positive cells were found in PVN after microinjection HRP into dorsal nucleus of vagal nerve. It proved that there was neural relationship between PVN and DVC. (3) The amplitude and frequency of gastric motility increased significantly by microinjection motilin into PVN in conscious rats. The effects produced by motilin could be abolished by vagotomy.

CONCLUSIONAll these results presented imply that motilin in PVN may increase gastric motility through PVN-DVC-Vagal nerve axis.

Animals ; Female ; Gastrointestinal Motility ; physiology ; Hypothalamus ; physiology ; Male ; Motilin ; physiology ; Paraventricular Hypothalamic Nucleus ; physiology ; Rats ; Rats, Wistar