Aim To explore the relationship between Mre11/Rad50/Nbs1 ( MRN ) complex focus formation and DNA double-strand breaks( DSBs) caused by cinob-ufagin in human hepatocellular carcinoma HepG2 cells. Methods The Na+,K+-ATPaseα1 subunit expression level in liver cancer tissues was detected by immunohis-tochemistry. After HepG2 cells were treated with 5μmol·L-1 cinobufagin for 6, 12 and 24 h, the drug-in-duced DSBs were assessed by single cell gel electro-phroesis ( SCGE ) , the gene transcription and protein levels of Mrel1, Nbs1, Rad50 and p53 were evaluated by Real time-PCR and Western blot. The cell cycle in parallel was analyzed by flow cytometry. Results The Na+, K+-ATPase α1 subunit expression level in liver cancer tissues was significantly increased compared with the tissue adjacent to carcinoma ( P <0. 05 ) . The 5μmol · L-1 cinobufagin could induce the DSBs in a time-dependent manner (P <0. 05), and it could up-regulate the gene expression levels of Mre11, Nbs1, Rad50 and p53 in HepG2 cells ( P<0. 05 ) . The pro-portions of HepG2 cells in S phase were ( 21. 32 ± 4. 21) % in the control group, and (33. 25 ± 5. 72) %, (56. 72 ± 6. 29) % and (67. 32 ± 9. 42) % in HepG2 cells treated with 5 μmol · L-1 cinobufagin for 6, 12 and 24 h, respectively. The proportions of cells in S phase in cinobufagin groups were significantly increased compared with the control group ( P<0. 05 ) . Conclu-sion Cinobufagin could induce the cell cycle arrest in liver cancer HepG2 cells by activation of Mre11/Rad50/Nbs1 Complex.

Objective To investigate the infection status of human papillomavirus (HPV) of females in Chongqing city ,in or‐der to provide scientific references for prevention of HPV infection and control of early cervical lesions .Methods The detection re‐sults of cervical cytology screening for 19 860 females in the University‐Town Hospital of Chongqing Medical University ,the First Affiliated Hospital of Chongqing Medical University and a county‐level hospital in Chongqing from January 2012 to January 2015 were collected ,and the HPV typing data were analysed .Results Among the 19 860 females ,5 681 females were infected with HPV ,the infection rate was 28 .61% .Due to the presence of multiple infection ,the actual infection frequency of HPV subtypes in 5 681 females was 8 906 .The infection frequency of high‐risk subtypes ,including multiple infections ,was 6 166 (accounted for 69 .23% ) .According to the probability of infection ,the top eight rate of subtype infection was HPV 52 ,16 ,58 ,6 ,CP8304(81) ,33 , 53 and 18 .The positive rate of single HPV infection was 19 .38% ,the positive rate of multiple infection was 8 .74% ,in which doub‐le infection was the main infection model .The infection rate of females aged 20 years old or less(43 .89% ) was the highest ,while have the infection rate of females aged 36-50 years old was the lowest(27 .26% ) .Conclusion The HPV infection rate of females in Chongqing is high ,the most of females infected by high‐risk subtype HPV .It is necessary to lay emphasis on preventing HPV 52 , 16 ,58 ,33 ,18 infection for females in Chongqing city .

Objective To study the consistency among non-invasive and invasive brachial artery pressure,radial artery pressure and invasive central arterial pressure,and to explore the correlation between the severe degree of coronary artery disease and invasive central aortic pressure.Methods A total of 331 patients who underwent coronary angiography in our hospital were selected.The invasive central aortic pressure,invasive and non-invasive brachial arterial pressure,radial artery pressure in all patients were measured.The severe degrees of atherosclerosis were recorded.The differences among invasive brachial arterial pressure and invasive radial artery pressure,non-invasive brachial artery pressure and non-radial artery pressure and invasive central aortic pressure were compared.Results The systolic pressure values measured in invasive and non-invasive brachial artery and radial artery were higher than that measured by central aortic pressure,while the diastolic pressure values measured in the four peripheral artery were lower than that measured in central aorta.The pressure values measured by non-invasive brachial artery pressure were more close to that measured by invasive central aortic pressure (P＞0.05).The systolic pressure was increased and the diastolic pressure was reduced in central aortic pressure with the coronary vessel lession numbers increased.The values of systolic pressure in patients with single-vessel,double-vessel and triple-vessel lesions were (118.2± 19.5) mm Hg,(124.9 ± 19.7) mm Hg and (137.7 ± 20.6) mm Hg,respectively and the values of diastolic pressure were (86.8±8.4) mm Hg,(85.3± 10.3) mm Hg and (83.1± 9.4) mm Hg,respectively.There were significant differences in systolic and diastolic pressure values among patients with single-vessel lesions,double vessel lesions and triple-vessel lesions(F=3.93,4.31,both P＜ 0.05).Conclusions The blood pressure values measured by noninvasive brachial artery pressure are more close to that measured by invasive central aortic pressure.There is a significant correlation between the severe degree of coronary heart disease and invasive central aortic pressure.Non invasive brachial artery pressure can be used in the early detection of cardiovascular dysfunction.

Objective To observe the clinical effect of Xiaoke-Yuzu decoction on diabetic peripheral neuropathy (DPN). Methods A total of 100 DPN inpatients were recruited and randomly divided into the treatment and control groups. The two groups were both received basic therapy, while the treatment group additionally received Xiaoke-Yuzu decoction. Toronto clinical scores and Chinese medicine symptom scores of both groups were collected to evaluate the clinical effect before and after the therapy. Results The Toronto scores of treatment group were significantly lower than control group after treatment (symptoms score 1.50 ± 0.94 vs. 2.23 ± 1.01, reflection score 3.60 ± 1.77 vs. 4.27 ± 1.72, feeling test score 1.53 ± 0.63 vs. 2.10 ± 0.84,all P<0.05). Meanwhile, the Chinese medicine symptom scores of treatment group were also significantly lower than the control group (main symptom score 1.77 ± 1.17 vs. 3.17 ± 1.82, posterior symptom score 2.23 ± 1.59 vs. 4.27 ± 1.57, the tongue and pulse score 1.83 ± 0.65 vs. 2.47 ± 0.51, all P<0.05). Conclusion Xiaoke-Yuzu decoction plus basic therpy could improve the clinical symptoms of DPN patients.

ObjectiveTo explore the correlation between butylphthalide and plasma brain-type creatine kinase isoenzyme (CK-BB),endothelin (ET),calcitonin gene related peptide (CGRP) in patients with acute cerebral infarction (ACI).MethodsSixty patients with ACI were divided into treatment group and control group with 30 cases each by random digits table method.Both groups received normal treatment continuously for 7 days,and butylphthalide of 200 mg was added to treatment group for 3 times per day.The levels of plasma CK-BB,ET and CGRP of two groups before and after treatment were measured and compared.ResultsThe levels of plasma CK-BB,ET of two groups both decreased significantly after treatment,but CGRP rose obviously.It showed significant differences before and after treatment of both groups (P ＜0.01 or ＜ 0.05).The levels of plasma CK-BB,ET of treatment group after treatment were lower than those of control group in the same period [(216.48±36.95) U/L vs.(333.07±54.03) U/L,(83.33±26.48)ng/L vs.(98.46±31.46) ng/L,P ＜ 0.05 ].The level of plasma CGRP of treatment group after treatment was significantly higher than that of control group in the same period [ (44.16±13.28 ) ng/L vs.(36.42±12.31 )ng/L,P ＜ 0.05 ].ConclusionButylphthalide can reduce the plasma enzyme activity,balance ET and CGRP,extend the cerebral arteries to antagonize ET,improve cerebral ischemia and cerebral hypoxia,which can protect brain cells and endothelial cells.

Objective To investigate the clinical prognostic value of neutrophils to lymphocyte ratio(NLR)in patients with ST-segment elevation myocardial infarction(STIMI)who underwent percutaneous coronary intervention(PCI). Meth?ods A total of 326 patients with acute ST segment elevation myocardial infarction were enrolled. According to the neutro?phils to lymphocyte ratio, patients were divided into group T1(≤3.46), group T2(3.47-6.75)and group T3(>6.75). Basic clinical data,laboratory indexes,coronary angiography results, TIMI grading and left ventricular ejection fraction(LVEF) were collected in all groups. Incidence of adverse events in hospital such as acute heart failure, thrombosis, ventricular tachy?cardia and ventricular fibrillation, atrial fibrillation, atrioventricular block and cardiogenic shock, were recorded. Major ad?verse cardiac events(MACE), death of all causes, non-fatal MI, ischemic stroke and readmission due to heart failure at the 6-month follow-up were also noted. Results The incidence of heart failure, atrial fibrillation, ventricular tachycardia/ven?tricular fibrillation and cardiogenic shock was higher in group T3 than those in the other two groups(P < 0.05).The inci?dence of multivessel coronary artery disease was significantly higher in group T3 than that in group T1 (54.1% vs 37.0%, P<0.05). The percentage of TIMI grade 3 in post-operative infracted artery was significantly lower than that in group T1 (83.5%vs 94.4%). Left ventricular ejection fraction was significantly lower in group T3 than that of group T1 during hospital?ization (0.48 ± 0.10 vs 0.52 ± 0.10). There was no significant difference in the incidences of death, non-fatal MI, ischemic stroke at 6 month follow up. But the rate of readmission due to heart failure was significantly higher in group T3 than that in the group T1(P<0.05). Conclusion Neutrophils to lymphocyte ratio may be a useful marker to predict the prognosis of pa?tients with heart failure due to acute myocardial infarction.

Objective To explore associations between SG13S114A/T and SG13S32A/C polymorphisms of ALOX5AP gene and the genetic susceptibility of ischemic cerebrovascular diseases (ICVD) in Henan Han population.Methods Two hundred and forty-six ICVD patients and 245 healthy controls were recruited from Han population in Henan province. Polymorphisms of SG13S114A/T and SG13S32A/C in ALOX5AP gene were genotyped in these samples by SnaPshot minisequencing method.Each genotype frequency and allele frequency were statistically analyzed and compared between ICVD group and control group using SPSS16.0 software.Haplotype and linkage disequilibrium were analyzed by SHEsis software.Results The SG13S114 AA genotype frequency ( 18.7％ ) and A allele frequency (41.3％) in ICVD group were significantly higher than those in control group (9.0％ and 32.7％,respectively; P =0.002 and P =0.005 ).It was also found that in male ICVD group and in younger ICVD group ( ＜50 years old),the SG13S114 AA genotype frequencies (22.1％ and 22.0％,respectively) and A allele frequencies (42.1％ and 42.7％,respectively) were significantly higher than those in male control group and younger control group (SG13S114 AA genotype:9.0％ and 8.9％ ; P =0.010 and P =0.006,respectively) ;A allele frequencies,34.0％ and 32.0％ ; P =0.048 and P =0.020,respectively.Finally,the prevalence of A-A haplotype in ICVD group was significantly higher than that in control group(30.4％ vs 23.5％,OR =1.419,95％ CI 1.068-1.885,P =0.015).T-C haplotype frequency of ICVD group was significantly lower than that in control group (22.0％ vs 28.8％,OR =0.698,95％ CI 0.523-0.932,P =0.014 ).Conclusions The A allele in SG13S114 loci of ALOX5AP may be a genetic risk factor for ICVD in Han population in Henan province.The association is predominant in ICVD patients of male and younger than 50 years old.Maybe A-A haplotype increases the risk of ICVD and T-C haplotype and has a protective effect against ICVD in Henan Han population.

Objective To investigate the role of TNSALP gene detection in prenatal diagnosis of HPP. Method The clinical data and the results of complete exon sequencing of TNSALP gene in one neonate with low alkaline phosphatase (HPP) were analyzed retrospectively. Peripheral bloods from his family members were collected. The amniotic fluid cell in fetuses at 17 weeks was tested for candidate gene mutations by Sanger sequencing. Results Mainly manifestations in 6-day-old baby were multiple fractures, limb shortening and bending and dyspnea. He died of respiratory failure 9 days after birth. The serum alkaline phosphatase was decreased and serum calcium was decreased slightly; serum phosphorus, serum 25 hydroxyvitamin-D and parathyroid hormone were normal. X-ray showed that the whole body bone was very poorly mineralized, and the long diaphysis was enlarged with shape of a cup at the end and multiple fractures existed. Gene sequencing revealed a complex heterozygous missense mutation in the TNSALP gene, including the heterozygous missense mutation c.542C>T in exon sixth causing 181st amino acids changed from serine to leucine (p.S181L), and tenth exon heterozygous missense mutation in c.1016G>A causing 339th amino acid changed from glycine to glutamic acid (p.G339E). The parental phenotypes were normal. The c.542C>T mutation is inherited from his father and the c.1016G>A mutation is inherited from his mother. These two mutations were not detected in the fetus. Conclusion TNSALP gene analysis can be applied to the diagnosis and prenatal diagnosis of HPP.

Objective Using single direction dispersion breathless DWI, to analyze the value of DWI for vertebral bone marrow infiltration in patients with acute leukemia (AL). MethodsForty-two patients with AL and 15 healthy volunteers received vertebral sagittal DWI with single shot spin-echo echoplan imaging (SS-SE-EPI) sequence( b value ＝ 0,650 s/mm2) at a GE Signa Excite 1. 5 T scanner with 8 channels body coil. DWI for all patients were performed from three directions, including from superior to inferior (S/I), from anterior to posterior (A/P) and from right to left (R/L). The apparent diffusion coefficient (ADC) value was measured on ADC map from each direction using GE-Function tool DWI software. Forty two patients consisted of 13 onset with untreated patients and 29 treated patients (7 nonremission,8 complete remission and 14 consolidation therapy). The ADC values among the three diffusion directions were compared. Analysis of variance and t test were used to compare the ADC values in different AL stages, Pearson correlation analysis was used to analyze the correlation between ADC values and the percentage of bone marrow progenitor cells. Results The ADC values from S/I, A/P and R/L of 362 vertebras in the 57 subjects are (0. 758 ±0. 009) × 10-3 mm2/s, (0. 732 ±0. 009) × 10 -3 mm2/s and (0. 758 ±0. 009) × 10 -3 mm2/s, respectively. There is no statistical significance( F ＝ 2. 958, P ＞ 0. 05 ).The ADC values from S/I of 94 vertebras in 15 healthy volunteers is (0. 697 ± 0. 122) × 10 -3 mm2/s, of 85 vertebras in 13 untreated AL patients is (0. 592 ±0. 071 ) × 10-3mm2/s. There is statistical significance between them ( t ＝ 2. 568, P ＜ 0. 05 ) ; The ADC value of 183 vertebras in 29 treated AL patients [ ( 0. 796 ±0. 225 ) × 10-3mm2/s]is higher than that in untreated patients with statistical significance (t ＝ -1. 332,P ＜0. 05). One hundred and forty vertebras in patients with complete remission and consolidation therapy were [ (0. 786 ±0. 184) × 10-3 mm2/s],and 43 vertebras in patients with non-remission(NR) [ (0. 804 ±0. 327 ) × 10 - 3 mm2/s], there was not statistical significance between them ( t ＝ - 0. 160, P ＞ 0. 05 ). The ADC values from S/I direction of untreated patients showed significant negative correlation with the proportion of the blast cell in the bone marrow ( median value 26. 4％. Min 7.9％, Max 48. 2％ ) ( r ＝- 0. 524, P ＜ 0. 05 ). ConclusionsDWI of vertebral bone marrow is isotropy. ADC value is a non-invasive and quantitative index for evaluating the pathogenetic condition of AL.

OBJECTIVETo study the protective effect of single prescription of traditional Chinese medicine Zige lyophilized powder on hypoxia/reoxygenation injury of human umbilical vein endothelial cells (HUVECs).

METHODHUVECs were cultured in vitro and Krebs liquid was adopted to establish the hypoxia/reoxygenation injury model. After intervention of Zige lyophilized powder, MTT colorimetric method was used to determine cell activity, the xanthine oxidase method is adopted to detect the activity of superoxide dismutases (SOD) in cells, the thibabituric acid developing method was adopted to determine the content of malondialdehyde (MDA), the nitrate reductase method was used to detect the content of nitric oxide (NO), the 2,4-dinitrophenylhydrazine developing method was adopted to the content of extracellular lactate dehydrogenase (LDH) , and the Western blot method was used to analyze the expression of cell apoptosis-related proteins Bcl-2, Bax and Caspase-3.

RESULTCompared with the model group, 12.5 mg x L(-1) of Zige lyophilized powder can enhanced cell viability (P < 0.05). Besides, 49.0, 24.5, 12.5 mg x L(-1) of Zige lyophilized powder prevents the decline of SOD activity during H/R (P < 0.05). Furthermore, 24.5, 12.5 mg x L(-1) Zige lyophilized powder significantly inhibited the increase of MDA and NO content and improved the expression of Bcl-2 protein (P < 0.05, P < 0.01). And 49.0, 24.5, 12.5 mg x L(-1) of Zige lyophilized powder significantly decreased Bax level and up-regulated Bcl-2/Bax ratio. Caspase activation and apoptosis were monitored in the reoxygenated cells. 49.0, 24.5 mg x L(-1) Zige lyophilized powder can decreased Caspase-3 expression (P < 0.01).

CONCLUSIONZige lyophilized powder can prevent H/R induced injury to HUVECs, which may be related to its effect in inhibiting cell peroxidation injury and enhancing cell antioxidant and antiapoptotic capacities

Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Cell Hypoxia ; drug effects ; Cells, Cultured ; Drugs, Chinese Herbal ; pharmacology ; Human Umbilical Vein Endothelial Cells ; cytology ; drug effects ; metabolism ; Humans ; L-Lactate Dehydrogenase ; metabolism ; Malondialdehyde ; metabolism ; Nitric Oxide ; metabolism ; Superoxide Dismutase ; metabolism ; bcl-2-Associated X Protein ; metabolism