Obective To evaluate the effects of dihydrotestosterone (DHT) on the expression of sterol regulatory element-binding protein-1c (SREBP-1c) in human HaCaT keratinocytes.Methods HaCaT cells were cultured in vitro and classified into 4 groups,i.e.,control group receiving no treatment,DIIT group treated with 3 different concentrations (10,100,1000 nmol/L) of DHT,LY294002 plus DHT group treated with DHT of 100 nmol/L after 40-minute pretreatment with the PI3K inhibitor LY294002 of 50 μmol/L,PD98059 plus DHT group treated with DHT of 100 nmol/L after 40-minute pretreatment with the MEK inhibitor PD98059 of 50 μmol/L.After another 24-hour culture,real time PCR and Western blot were carried out to detect the expression of SREBP-1c mRNA and protein in HaCaT cells,respectively.Western blot was also performed to determine the phosphorylation levels of protein kinase B (AKT),extracellular signal-regulated kinase (ERK),p38 mitogen activated protein kinase and c-Jun N-terminal kinase (JNK) in the HaCaT cells.Results DHT could enhance the expression of SREBP-1c mRNA and protein in HaCaT cells in a concentration-dependent manner,and induce the phosphorylation of AKT and ERK,but not that of P38 or JNK.The expressions of SREBP-1c mRNA and protein were significantly decreased in HaCaT cells treated with LY294002 plus DHT (7.4780 ± 1.2638 vs.21.6170 ± 2.2759,t =9.406,P ＜ 0.05; 0.7113 + 0.0313 vs.2.2577 + 0.0601,t =39.498,P ＜ 0.05),but experienced no statistical changes in those treated with PD98059 and DHT(both P ＞ 0.05),compared with those treated with DHT only.Conclusion DHT can induce the expression of SREBP-1c mRNA and protein in HaCaT cells,likely via the PI3K/AKT signaling pathway.

Objective Gasp was defined as a pathology respiration during cardiac arrest. This study was to investigate its effect on hemodynamics during CPR. Method Twelve domestic pigs, weighening (30 ± 1) kg,were anaesthetized. After tracheal intubation and mechanical ventilation, continuous respiratory variables were recorded. An artery catheter was inserted for reference blood samples and measuring aortic artery pressure (AOP).Right atrial pressure (RAP) and cardiac output (CO) were detected by Swan-Ganz catheter. Ventricular fibrillation (VF) was induced by programmed electrical stimulation instruments. After 4 minutes untreated VF, standard 30:2 CPR was done for 12 minutes and the parameters were recorded. Results pH, PaCO2 and lactic acid increased and PaO2 decreased progressively during CPR, whereas PaO2 was up to 50mmHg during the whole protocol. Gasps were observed in 10 animals, but weaken gradually; the left 2 animals with no gasp did not restore of spontaneous circulation (ROSC). Standard CPR could produce passive ventilation more than dead space (VD), but its tidal volume decreased gradually, which led to the percentage of rescue ventilation increased progressively. Positive correlations were found between CO, coronary perfusion pressure (CPP) and minute ventilation of gasps (MVg) (r was 0.736 and 0.721 respectively, both P ＜0.01); negative correlation were found between RAP and MVg (r= -0. 744, P ＜ 0.01). Conclusions Standard CPR could maintain 12 minutes oxygenation of body; compressions could produce enough passive ventilation more than VD; gasps were benefit to ROSC by increasing CO, CPP and decreasing RAP.

Objective To investigate the relationship between the serum concentrations of brain derived neurotrophicfactor (BDNF) and its G196A polymorphism in the amphetamine induced-psychosis inpatients.Methods The cross-sectional study included 233 amphetamine abuses and 110 healthy participants who served as controls.The serum concentration of BDNF was measured by sandwich ELISA,and the genotype of BDNF G196A polymorphism was determined used polymerase chain reaction (PCR) techniques.The data were analyzed using SPSS 12.0 statistics software.Results The serum concentration of BDNF in case group((205.81±75.36) pg/ml) were significantly higher than that in control group((95.04±31.63) pg/ml;t=15.02,P＜0.01).There was no significant difference about the BDNF serum concentrations between the inpatients with the amphetamine induced psychosis and the inpatients with the amphetamine abuse (P＞0.05).The BDNF serum concentration showed no significant difference in the genotype distributions and allele frequencies (P＞0.05).The genotype distributions and allele frequencies of BDNF G196A showed no significant difference among three groups(P＞0.05).Conclusion The BDNF serum concentration is correlated with methamphetamine abuse,while the BDNF G196A gene polymorphism may not be associated.

Objective To investigate the effects of arsenic trioxide(ATO) on anti-dsDNA antibody and lymphocyte subsets proliferation in splenic cells of MRL/lpr mice.Methods MRL/lpr mice were divided into three groups: ATO group(0.4 mg/kg?d,ip),sodium chloride(NS) group(0.25 mL,ip),and cyclophosphamide(CTX) group(50 mg/kg?qw,ip).The control group consisted of 12 syngeneic normal C57/BL mice,which were sub-divided into ATO group(0.4 mg/kg?d,ip) and NS group(0.25 mL,ip).After two-month treatment,all mice were killed and their bodies and spleens were weighed.Anti-dsDNA antibody in serum were detected by ELISA.The percentage of different cell subsets was checked through flow cytometry.Results(1)The spleen index、serumlevel of anti-dsDNA antibody and the percentages of CD19+、CD3+、CD3+CD4+ lymphocytes in ATO group were much lower than those in NS group of MRL/lpr mice(P0.05). Conclusion Arsenic trioxide can inhibit the activation of T and B lymphocytes,increase the percentage of NK cell and reduce the serum level of auto-antibody.And it was of no effect in the normal mice.

Objective To analyze the complete genome sequence of Guangxi HEV isolate swGX32 and to compare it with other HEV isolates. Methods The overlapping fragments of HEV isolate swGX32 were amplified with reverse-transcription nested polymerase chain reaction (RT-nPCR),and the 5′ and 3′ ends of viral genome were amplified with rapid amplification of cDNA ends (RACE). The PCR products were cloned and sequenced. The sequence and phylogenetic analysis of swGX32 was performed. Results The genome of swGX32 consisted of 7240 nt excluding the polyA tail, with 4 nt overlapping between ORF1 and ORF2. ORF3 is contained in the sequence of ORF2. The complete genome sequence of swGX32 shared identity of 73%-74%, 73%, 74%-75%,83%-94% with HEV genotype 1,2,3 and 4, respectively. Among all these HEV reference sequences, swGX32 showed the highest identity with the human isolate JKO-ChiSai98C (94%). Phylogenetic tree showed that swGX32 belonged to genotype 4 and clustered with JKO-ChiSai98C in the branch of HEV subtype 4a. Conclusion The swine HEV isolate swGX32 is closely related to human strain JKO-ChiSai98C genetically and phylogenetically, which further provides molecular biology evidence of hepatitis E as a zoonosis.

Objective:To explore the expression of mammalian target of rapamycin (mTOR)and its relationgship with the prognosis of breast cancer,and to provide evidence-based basis for the using of mTOR inhibitor in the treatment of breast cancer.Methods: A systemical literature search was conducted based on the following databases:PubMed,EMBbase,Cochrane Library,ISI Web of Science,and CNKI up to November 24,2015.The outcome measures were hazard ratio (HR)with 95% confidence interval (CI) for the association between the mTOR expression and the prognosis of patients with breast cancer.The primary end points including disease-free survival (DFS ), and overall survival (OS ). STATA 12.0 was used to conduct the statistical analysis. Results:A total of seven cohort studies,1 758 patients were included. The risk of recurrence and metastasis of the breast cancer patients with positive expression of mTOR was 2.05 times of the patients with negative expression of mTOR (HR= 2.05, 95% CI: 1.01 - 4.13,P = 0.003);the risk of death in the breast cancer patients with positive expression of mTOR was 2.63 times of the patients with negative expression of mTOR (HR = 2.63, 95%CI:1.45-4.80,P = 0.736).Conclusion:The positive expression of mTOR can significantly increase the recurrence,metastasis and death risk of the patients with breast cancer.

Objective:To investigate the effects of hematopoietic progenitor kinase 1 (HPK1) overexpression by construction of lentiviral vector on the proliferation and apoptosis of breast cancer MCF-7 and MDA-MB-231 cells,and to elucidate its possible mechanism.Methods:The cells were infected with the lentivirus overxpressing HPK1,and the MCF-7-HPK1 and MDA-MB-231-HPK1 cell lines were stably expressed HPK1;each cell line was divided into three experimental groups:blank group (untreated),control group (empty vector) and HPK1-overexpression group.The expression levels of HPK1 mRNA and protein in breast cancer cells in each group were detected by RTPCR and Western blotting methods,respectively.The cell proliferation rate was detected by MTT assay.The cell cycle and apoptotic rate were detected by flow cytometry.Transwell assay was used to analyze the cell migration ability.Western blotting method was used to measure the expression levels of caspase 3,PTEN,MMP-9,MMP-2,Ki-67and HPK1 proteins.Results:Compared with blank groups and control groups,the expression levels of HPK1 mRNA and protein in the both cell lines in HPK1 overexpression groups were significantly up-regulated (P＜0.05),the proliferation rates were significantly decreased (P＜0.05) and the apoptotic rates were significantly increased (P＜0.05),the number of cells crossing matrigel was significantly reduced (P＜0.05),the cell cycle of MCF-7 was blocked in G1 phase (P＜0.05),the expression levels of caspase 3 and PTEN proteins in HPK1 overexpression group were significantly increased (P＜0.05),and the expression levels of MMP-2 and MMP-9 proteins were significantly decreased (P＜0.05).Conclusion:HPK1 overexpression can inhibit the proliferation and migration of MCF-7 and MDA-MB-231 cells and induce apoptosis,which may be related to the up-regulation of caspase 3 and PTEN and down-regulation of MMP-9,MMP-2 and Ki-67.

Objective To analyze the expression and role of ubiquitin specific peptidase 18 (USP18) in gastric cancer cells,and to investigate the relationship between the development of gastric cancer and USP18.Methods The levels of USP18 protein and mRNA expression in immortalized gastric mucosa epithelial GSE cell lines and gastric cancer cell lines (AGS,MKN45,MKN25,BGC823,BGC803,SGC7901) were detected by using reverse transcription-polymerase chain reaction (RT-PCR) and Western blot,respectively.The role of USP18 in the invasion and proliferation of gastric cancer cells was analyzed by using CCK8 and Transwell assays.Results The mRNA level of USP18 was lower in GSE cell lines than that in gastric cancer cells (F =794.052,P ＜ 0.000 1).In six gastric cancer cell lines,mRNA level of USP18 was relatively high in BGC823 (17.62±0.55) and BGC803 (13.52±0.50) cell lines,and low in MKN28 (1.40±0.17) and MKN45 cell lines (4.23±0.26).As for the protein level,the expression of USP18 was lowest in GSE cell line.In six gastric cancer cell lines,the expression of USP18 was the highest in more aggressive SGC7901 and BGC803 cell lines and the lowest in AGS and MKN45 cells.Compared with the control group,interference of USP18 decreased the invasion and proliferation abilities of SGC7901 and BGC803 cell lines (P ＜ 0.01).Conclusion USP18 is highly expressed in more invasive gastric cancer cells,and the downregulation of USP18 can suppress the invasion and proliferation of gastric cancer cells.

Objective To compare the changes in IFN-γ, IL-4, IL-10 and IL-1β expression at mRNA level in gastric mucosae of BALB/c, C57BL/6 and nude mice at different stages of Helicobacter heilmannii ( H. heilmannii) infection, and to investigate the types of induced immune responses. Methods Each kind of mice was randomly divided into two groups: infection ( n=30 ) and control ( n=6 ) groups. Those in the infection groups were intragastrically inoculated with H. heilmannii strains to establish long-term stable mouse infection models. Gastric mucosa tissues were collected at weeks 4, 8, 12, 24 and 36 and ana-lyzed by semi-quantitative RT-PCT to detect the expression of IFN-γ, IL-4, IL-10 and IL-1βat mRNA lev-el. Results In the early stage of infection (weeks 4-12), INF-γ, IL-4, IL-10, and IL-1β expression at mRNA level in the gastric mucosae of BALB/c and C57BL/6 mice were significantly increased compared with those of the control groups (P<0. 05). IL-4, IL-10 and IFN-γexpression peaked at weeks 8-12, while IL-1βexpression reached the peak at week 4. After 12 weeks, IFN-γexpression at mRNA level in BALB/c mice was significantly decreased, but showed no significant change in C57BL/6 mice. IL-4 expression at mRNA level in C57BL/6 mice at the late stage of infection (week 36) was lower than that in the correspond-ing control group (P<0. 05). Expression of IFN-γ, IL-4, IL-10 and IL-1β at mRNA level in nude mice were all higher than those in the control group, and there were significant differences in IL-1βand IL-4 ex-pression between groups (P<0. 05). Conclusions Expression of cytokines in H. heilmannii-infected mice increased over time. IFN-γ-mediated Th1 immune responses were the predominant immunity induced by H. heilmannii infection. Immune responses to H. heilmannii infection varied with the kinds of mice. C57BL/6 mice showed mainly Th1 cell immune responses, while Th1/Th2 mixed immune responses were induced in BALB/c mice.

OBJECTIVETo investigate cardiac function and myocardial perfusion during 48 h after cardiopulmonary resuscitation (CPR), further to test myocardial stunning and seek indicators for long-term survival after CPR.

METHODSAfter 4 min of untreated ventricular fibrillation, fifteen anesthetized pigs were studied at baseline and 2 h, 4 h, 24 h, and 48 h after restoration of spontaneous circulation (ROSC). Hemodynamic data, echocardiography and gated-single photon emission computed tomography myocardial perfusion images were carried out.

RESULTSMean arterial pressure (MAP), coronary perfusion pressure (CPP) and cardiac troponin I (CTNI) showed significant differences between eventual survival animals and non-survival animals at 4 h after ROSC (109.2 ± 10.7 mmHg vs. 94.8 ± 12.3 mmHg, P=0.048; 100.8 ± 6.9 mmHg vs. 84.4±12.6 mmHg, P=0.011; 1.60 ± 0.13 ug/L vs. 1.75 ± 0.10 ug/L, P=0.046). Mitral valve early-to-late diastolic peak velocity ratio, mitral valve deceleration time recovered 24 h; ejection faction and the summed rest score recovered 48 h after ROSC.

CONCLUSIONCardiac systolic and early active relaxation dysfunctions were reversible within survival animals; cardiac stunning might be potentially adaptive and protective after CPR. The recovery of MAP, CPP, and CTNI could be the indicators for long-term survival after CPR.

Animals ; Blood Pressure ; Cardiopulmonary Resuscitation ; Coronary Circulation ; Heart Arrest ; Hemodynamics ; Male ; Myocardial Contraction ; physiology ; Myocardial Stunning ; Swine ; Time Factors ; Ventricular Fibrillation