Objective To observe the activation of rat hepatic stellate cells (HSC-T6) and the changes of methyltransferase (DNMT)1,DNMT3a mRNA expression with different doses of sodium arsenite stimulation.Methods HSC-T6 cells were exposed to a final concentration of 0 (control),5 (low dose),15 (medium dose) and 25 (high dose) μmol/L sodium arsenite in culture medium for 24,48 and 72 h,cells and cell culture supernatant were harvested.Enzyme-linked immunosorbent assay (ELISA) kit was used to measure fibrosis factors contents of type Ⅰ collagen (COL-1),type 11Ⅲ collagen (COL-3) and alpha smooth muscle actin (α-SMA) and quantitative real-time PCR was used to detect the expression levels of DNMT1 and DNMT3a mRNA.Results Different arsenic exposure time (24,48,72 h) had a significant effect on COL-1,COL-3 and α-SMA contents in HSC-T6 cells (F =249.574,328.493,3 157.436,all P ＜ 0.01);Different arsenic exposure content (low,medium,high dose groups) had a significant effect on COL-1,COL-3 and α-SMA contents in HSC-T6 cells (F =3 946.521,1 006.399,13 025.770,all P ＜ 0.01).After arsenic exposure for 24 and 48 h,the expression levels of DNMT1 mRNA in high dose group (4.33 ± 0.24,2.34 ± 0.43) were higher than those of control group (1.00 ± 0.00,1.00 ± 0.00,all P ＜ 0.05).At the same arsenic exposure levels (low,medium or high dose),the expression level of DNMT1 mRNA was declined with prolongation of sodium arsenite stimulation time (all P ＜ 0.05).After arsenic exposure for 48 and 72 h,the expression levels of DNMT3a mRNA in high dose group (2.23 ± 0.50,5.02 ± 0.23) were higher than those of control group (1.00 ± 0.00,1.00 ± 0.00,all P ＜ 0.05).The expression levels of DNMT3a mRNA in medium and high dose groups at 72 h (3.80 ± 0.14,5.02 ± 0.23) were higher than those of 24 h (3.03 ± 0.12,0.42 ± 0.15,all P ＜ 0.05).Conclusion HSC-T6 cells are obviously activated with pro-fibrotic effect;the expression levels of DNMT1 and DNMT3a mRNA are both up regulated in HSC-T6 cells after being exposed to sodium arsenite.

OBJECTIVE:To establish a method for the content determination of geniposide in TCM callus convenient wipes drug solution. METHODS:HPLC was performed on the column of Agilent Zorbax SB-C18 with mobile phase of acetonitrile-0.1%phosphoric(10:90,V/V)at the flow rate of 1.0 ml/min,column temperature was 30 ℃,detection wavelength was 236 nm and the volume was 10 μl. RESULTS:The linear range of geniposide was 0.104 1-1.041 μg (r=0.999 8);RSDs of precision,stability and reproducibility tests were all lower than 2%;recovery was 99.04%-100.82%(RSD=0.85%,n=6). CONCLUSIONS:The method is simple,accurate,reproducible,and can be used for the content determination of geniposide in TCM callus convenient wipes drug solution.

Objective To study the mechanism of liver fibrosis in rats caused by chronic exposure through drinking water containing sodium arsenite,to identify the differential proteins via proteomics technique.Methods Totally 40 healthy 8-week-old male Sprague Dawley (SD) rats of specific pathogen free (SPF) grade were randomly divided into 4 groups,which were control group (deionized water),0.68,1.36 and 2.73 mg/kg sodium arsenite (iAs3+) treated groups,respectively.The rats were fed with iAs-treated drinking water freely for 24 consecutive weeks.Twenty-four hour urine sample,blood and liver samples were collected.Hepatic fibrosis indices,specifically,type Ⅲ precollagen (PC Ⅲ),type Ⅳ collagen (Ⅳ-C),hyaluronic acid (HA) and laminin (LN) were detected by enzymelinked immunoassay (ELISA).Based on the isobaric tags for relative and absolute quantitation (iTRAQ) reagent 8-plex experiment,combined with 2DLC-MS/MS,the proteins in rats liver tissue of the medium dose group and the high dose group were compared with the those of control groups.Results ①The serum HA contents in the C (control) group,the L (low dose) group,the M (medium dose) group and the H (high dose) group were (198.51 ± 16.64),(218.39 ± 34.98),(261.72 ± 30.56) and (297.31 ± 35.72) ng/L;the serum PCⅢ contents in C,L,M and H groups were (15.32 ± 2.15),(16.78 ± 2.64),(19.51 ± 0.85) and (21.42 ± 1.63) μg/L;the serum LN contents in C,L,M and H groups were (734.57 ± 86.00),(792.65 ± 94.15),(916.83 ± 84.40) and (1 008.09 ± 64.17) μg/L;the serum Ⅳ-C contents in C,L,M and H groups were (52.34 ± 14.65),(59.72 ± 12.84),(74.38 ± 4.83) and (78.46 ± 4.30) μ.g/L,respectively.The differences in serological indices of liver fibrosis between-groups were statistically significant (F =21.136,19.957,22.007,14.288,all P ＜ 0.05).In multiple comparison of serum HA,PCⅢ and LN,there were no statistical significant differences between L group and C group.M and H groups were higher than L group and C group,significant statistical difference was found between H group and M group (all P ＜ 0.05).②Combining iTRAQ with 2DLC-MS/MS,based on the confidence threshold of protein (unused protScore) ＞ 1.3 and at least 1 matched peptides within the 95％ confidence interval,2 948 proteins were identified.Totally 2 162 proteins were detected in three groups compared with Venn diagram,after removing significant different proteins in C group,687 up-regulated proteins and 548 down-regulated proteins were identified in M group;633 up-regulated proteins and 519 downregulated were found in H group;the differences of protein expression between M and H groups were not statistically significant (P ＞ 0.05).③Up-regulated proteins related to the metabolism including AS3MT,MAT,SHMT,CHDH,CTH,CSAD and BHMT in M and H groups;of the two kinds of proteins of MTR,METK1 was up-regulated and F1LRB8 was down-regulated.Proteins associated with GSH including Gsta1,Gsta4,Gsta5,Gstt1,Gstt2,Gstk1,Gstp1,Gstm1,Gstm2,Gstm3,Gss,Gpx1,Gpx4,Esd,Hagh,Glo1,Mgst1 and B6DYQ5 which were all up-regulated.Proteins associated with liver fibrosis were Hic-5,Gss and six kinds of Tpm,and six kinds of Tpm subunits including two kinds of Tpm1,three kinds of Tpm2 and one kind of Tpm3 which were all up-regulated.Conclusions There is liver accumulation of arsenic after chronic arsenic exposure and resulting in liver fibrosis and decline of liver function.Expressions of AS3MT,MTR,MAT,SHMT,BHMT,CHDH,CTH and CSAD are up-regulated;arsenic meta bolism methionine cycle,folic acid cycle and sulfur transfer pathways are closely related.GSH plays an important role in arsenic metabolism and liver fibrosis,Hic-5,GSS and TPM may be associated with the occurrence of liver fibrosis.

Objective To investigate the expression of Fc?RⅡb on peripheral blood mononuclear cells(PBMCs)and serum anti-C1q antibodies in SLE patients and their role in SLE pathogenesis.Methods The ex-pression of Fc?RⅡb on peripheral neutrophiles,lymphocytes and monocytes was detected by flowcytometry and the level of anti-C1q antibodies was tested by ELISA in32SLE patients and22healthy individials.At the same time,the correlation between Fc?RⅡb,anti-C1q antibodies and ANA,anti-dsDNA antibodies,SLEDAI was eval-uated respectively.Results The expression of Fc?RⅡb on peripheral neutrophiles,lymphocytes and monocytes(especially on neutrophiles and monocytes)of SLE patients decreased,the level of anti-C1q antibodies was signifi-cantly increased,compared with that of the control group.Fc?RⅡb was negatively and anti-C1q antibodies were positively associated with ANA,anti-dsDNA antibodies and SLEDAI respectively.Conclusion The defective ex-pression of Fc?RⅡb on PBMCs and high level of anti-C1q antibodies do exist in SLE patients.Fc?RⅡb and anti-C1q antibodies may decrease the clearance of immune complex and play an important role in the pathogene-sis of SLE.They may also be important parameters in indicating SLE activity.

The effects of quercetin-3-O-β-D-glucuronide (Q3GA)on the triglyceride metabolism and oxidative stress in steatotic HepG2 cells and the underlying mechanism were investigated in this study.Significant fat accu-mulation was documented by Oil Red O staining;intracellular triglyceride levels were detected by triglyceride(TG)enzymatic assay.DCFH-DA staining assay was performed to observe reactive oxygen species (ROS)pro-duction of HepG2 cells.The level of malondialdehyde(MDA)and superoxide dismutase(SOD)were assayed by thibabituric acid method and xanthine oxidase method.Changes in the mRNA expression of peroxisome prolifera-tor-activated receptorα(PPARα);carnitine palmitoyltransferase 1 A(CPT1 A);medium chain acyl-CoA dehydro-genase (MCAD);cytochrome P450 4A11(CYP4A11)and acyl-CoA oxidase(ACO);which are related with fatty acid oxidation were assessed by RT-PCR.Our results showed that Q3GA obviously reduced fat deposition and TG content.At the same time;Q3 GA decreased MDA content and significantly increased the SOD activity with reduced ROS production.Moreover;the PPARα;CPT1A;MCAD expression-related fatty acid βoxidation was upregulated with the treament of Q3GA;while without any change of the expression of CYP4A11;ACO.In conclu-sion;Q3GA prevents FFA-induced HepG2 cell steatosis;and enhances mitochondrial fatty acidβoxidation;which may partly be related to its anti-oxidation ability.

Objective To compare the risk factors of breast cancer in Ningxia Hui and Han women,and to provide evidence for the prevention and control of breast cancer.Methods The female patients of breast cancer treated at the Affiliated Hospital of Ningxia Medical University between May 2013 and July 2014 were chosen for case study,while other patients treated at the same hospital and during the same period who did not have breast cancer were selected as a control. An epidemiological survey was conducted using the same questionnaire among the two groups.The survey involved general demographic information,menstrual history,reproductive history,life habit and family history of cancer.Risk factors of breast cancer in Hui and Han nationalities were analyzed by logistic regression analysis.Results Logistic regression analysis showed that the abortion number(OR =2.631,P =0.028)was a risk factor for the occurrence of breast cancer in women of the Hui nationality,while physical exercise (OR =0.177,P =0.040)was a protective factor.Tumor suffered by immediate family members (OR =4.249,P =0.014),abortion number (OR =1.602,P =0.001 ),the age of the first childbirth (OR =1.253,P =0.001 )and the age of first marriage(OR =1.223,P =0.001 )were the major risk factors while physical exercise (OR =0.422,P =0.001 )was a protective factor against breast cancer in Han nationality. Conclusion The risk factors of breast cancer in the women of Hui and Han nationalities are consistent in terms of the total number of abortions and physical exercise.Compared with the Hui people,the age of first marriage,the age of first child birth,and tumor suffered by immediate family members also play a role in the occurrence of breast cancer in the Han na-tionality.

OBJECTIVE To investigate the protective effect of Sika deer velvet antler protein (SVPr) against renal toxicity in mice and its mechanism.METHODS Forty ICR mice were randomly divided into 5 groups:normal control group (ig distilled water),model group (ig distilled water for 7 d,on the 7th day,ip cisplatin 25 mg·kg-1 to establish the model,afterwards ig distilled water for 3 d) and SVPr 5,10 and 20 mg· kg-1 groups (ig SVPr for 7 d,cisplatin 25 mg· kg-1 was provided 2 h after the last administration,then ig SVPr for 3 d).Testing kits were adopted for the measurement of renal indexes in mice,such as blood urea nitrogen (BUN) and serum creatinine (SCr);oxidative stress indictors of super oxide dismutase (SOD),catalase (CAT),glutathione (GSH) and malondialdehyde (MDA);inflammation indictor levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6).Caspase 3,Bax and Bcl-2 were detected via Western blotting,and renal pathological changes were observed by HE staining.RESULTS SVPr (5,10 and 20 mg·kg-1) significantly reduced the levels of SCr,BUN,MDA,TNF-α and IL-6,and the expressions of caspase 3 and Bax (P＜0.05),but increased the activities of SOD,CAT and GSH,and the expression of Bcl-2 (P＜0.05).The renal pathological changes were improved.CONCLUSION SVPr can reduce renal toxicity induced by cisplatin in mice,and the mechanism is probably related to inhibiting oxidative stress or inflammatory reaction and improving cell apoptosis.

ABSTRACT:Objective To study the effect of curcumin on the learning and memory ability in a rat model of Alzheimer’s disease (AD).Methods AD rat model was prepared using intraventricular injection of Aβ1-42. Curcumin was acutely (single injection before the behavioral tests)or chronically (injected for 6 consecutive days) injected intraperitoneally at doses of 50,100 or 300 mg/kg.Brain-derived neurotrophic factor (BDNF)protein (1 μg/side)or BDNF shRNA (2×10 5 units/side)was infused into the hippocampus.The behavioral changes in Y-maze,open field test and Morris water maze and the expression of BDNF in the hippocampus were analyzed. Results Acute treatment with curcumin had no significant effects on the spontaneous alteration,locomotor activity or water maze latency of AD rats.AD rats treated chronically with curcumin (300 mg/kg ) showed significant elevation in the spontaneous alternation (P <0.000 1)in Y-maze and memory ability in the water maze test (P <0.05 )compared with those in the saline group.Chronic treatment with 100 and 300 mg/kg of curcumin induced an increased level of BDNF in the hippocampus as compared with the saline controls (P <0.05 and <0.000 1). Intrahippocampal injection of BDNF significantly decreased the escape latency of AD rats in the water maze (F 4,2 9 5=5.813,P <0.01 ).Rats chronically injected with curcumin combined with shBDNF showed no difference in the swimming time in Ⅱ quadrant as compared with saline controls (P =0.657).However,rats in 100 mg/kg curcumin group,BDNF group and sham group had significantly increased swimming time than the saline controls (P <0.05, P <0.05 and P <0.000 1,respectively).Conclusion Curcumin may activate the downstream signaling pathways by upregulating the expression of BDNF and ultimately contribute to the improvement of learning and memory in AD rats.

Objective To evaluate the ultrasonic feature and clinical significance of persistent left superior vena cava(PLSVC)in fetal life.Methods Fetal echocardiography was performed in 3368 fetuses.Thirty-one fetuses of PLSVC were confirmed.Results The dilated coronary sinus was observed in 30 of 31 fetuses.Congenital heart defects were presented in 14 of these cases,and extracardiac anomalies were presented in 6 fetuses.Both congenital heart defects and extracardiac anomalies were observed in 4 fetuses.Conclusions PLSVC is always associated with congenital heart defects.The prognosis Of affected fetuses largely depends on whether or not the PLSVC is associated with a congenital heart defect.Prenatal diagnosis of PLSVC can help US plan perinatal counseling and ameliorate the postnatal course.

Objective To explore the effect of electroacupuncture at Quchi (LI11), Zusanli (ST36) on differentiation of neural stem cells after cerebral ischemia-reperfusion in rats. Methods Thirty-six male Sprague-Dawley rats were randomly divided into sham group (n=12), model group (n=12) and electroacupuncture group (n=12). The latter two groups were occluded the left middle cerebral arteries for 90 min-utes and reperfused. The electroacupuncture group received electroacupuncture at Quchi and Zusanli acupoints for 21 days. They were evalu-ated with modified Neurological Severity Scores 7, 14 and 21 days after electroacupuncture. Their infarct volumes were tested with MRI T2WI 21 days after electroacupuncture, while the differentiation of neural stem cells was observed with double-immunopositive BrdU/Dcx and BrdU/NeuN. Results Compared with the model group, the neurological deficits score improved in the electroacupuncture group in all the time points (P<0.05). The infarct volumes decreased in the electroacupuncture group (P<0.05), with less number of BrdU+/Dcx+cells in subventricular zone (P<0.001) and more number of BrdU+/NeuN+ cells in peri-infarct cortex (P<0.001) 21 days after electroacupuncture. Conclusion Electroacupuncture at Quchi and Zusanli acupoints can improve neurological function and decrease the infarct volumes in rats after cerebral ischemia-reperfusion, which may be associated with promoting differentiation of neural stem cells to neurons.