Objective To detect 5′CpG island methylation of APC(adenomatous polyposis coli) gene promoter region in cervical carcinoma cell lines (HeLa,CaSki,SiHa), and to investigate the possibility of Hydralazine in restoring the expression of APC gene through demethylation. Methods The CpG island methylation status of APC gene promoter region of the cervical carcinoma cell lines were analyzed using methylated specific polymerase chain reaction (MSP). The mRNA expression profile of APC gene was analyzed by using RT-PCR after Hydralazine treatment. Results The CpG island of APC gene was methylated in HeLa and hemimethylated in CaSki but not in SiHa. The mRNA expression of APC gene can be detected after hydralazine treatment. Conclusion The APC gene non-expression in cervical carcinoma is associated with CpG island methylation in APC gene promoter region. Hydralazine, served as a demethylating agent, enables to restore the expression of APC gene.

Objective To investigate the expression of survivin, matrix metalloproteinase (MMP-2) and tissue inhibitor 2 of matrix metalloproteinase (TIMP-2) in cervical carcinoma and their relationship with invasion and node metastasis of the cervical cancerous tissues. Methods The expressions of survivin, MMP-2 and TIMP-2 were examined by immunohistochemical S-P method and colour pathological image computer analysis system in 10 cases of normal cervical epithelia, 10 cases of cervical carcinoma in situ, 40 cases of invasive squamous cell cervical carcinoma and 11 cases of invasive cervical adenocarcinoma. The relationship between those indexes and the factors related to clinical pathology of cervical carcinoma were analyzed statistically. Results It was found that the positive level of survivin and MMP-2 expression increased in the order of normal cervical epithelium, cervical carcinoma in situ and invasive carcinoma of cervix (P0.05). The positive expressions of survivin and MMP-2 in patients under 35 years old or with pelvic lymph node metastasis, intravascular involvement and stroma involvement were significantly higher than that in the cases without them, while TIMP-2 expression was opposite to that of MMP-2 (P

Objective To investigate the effect of Calponin-1 suppression on human myometrium cells through adenovirus mediated siRNA. Methods Human uterine smooth muscle tissues were digested with enzymes, cultured and confirmed with immunocytochemistry. Aadenovirus siRNA-Calponin-1 plasmid was transfected into primary cultured uterine smooth muscle cells in vitro. The expressions of Calponin-1 mRNA and protein were analyzed by RT-PCR and Western blot, respectively.Results The pAdEasy-pShuttle-U6-Calponin-1 siRNA plasmid was successfully constructed, and Calponin-1 siRNA mediated by recombinant adenovirus resulted in markedly reduced expression of Calponin-1 mRNA and protein in human myometrium cells. The gray values of Calponin-1 mRNA in the uterine smooth muscle cells in the experimental, blank control, and empty vector groups were 316.3±39.2, 1048.5±126.4 and 1027.2±127.5, respectively. The gray values of Calponin-1 protein were 323.3±43.2, 1021.5±143.4, and 1019.2±144.5,respectively. The difference between the experimental group and the blank control group as well as the empty vector group was significant (P< 0.05). There was no significant difference between the empty vector group and the blank control group (P>0.05).Conclusion The pAdEasy-pShuttle-U6-Calponin-1 siRNA plasmid can inhibit the expression of Calponin-1 in human myometrium cells in vitro,which may be a useful approach to determine the role of Calponin-1 in delivery.

Objective To investigate the growth inhibition and radiosensitization of Celecoxib in hu-man nasopharyngeal carcinoma cell line CNE-2. Methods CNE-2 growth inhibition by Celecoxib was eval-uated by MTT method. Apoptosis-related changes in morphology were observed by transmission electron mi-croscopy (TEM). Cell cycle distribution and apoptosis rate were measured by flowcytometry (FCM). The ex-pression of COX-2 protein was observed by SP method after the treatment of Celecoxib. Cells were randomly planted into four groups: irradiation control(Ci), drug group(Cd), irradiation group(R), and Celecoxib plus irradiation group(D+R). Single irradiation of 2,4,6,8,and 10 Gy were administered for colonogenic assay. Cell cycle distribution and apoptosis rate were analyzed at 6 Gy irradiation. Results The growth of CNE-2 cell was inhibited by celecoxib in a dose-and time-dependent manner, the IC50 was 80 μmol/L After the treatment, cell ratio of GO and G, phases was increased (47.03±2.76 vs 56.17±1.95, t=4.68, P= 0.010), whereas the ratio of S and G2/M phases was decreased (33.07±1.86 vs 24.87±1.76, t=5.54, P = 0.010; 19.30±0.53: 17.73±0.83, t=2.75, P=0.050), and the apoptosis rate was increased (1.57±0.47:10.47±0.31, t = 27.39, P = 0.000) in a dose-dependent manner. Apoptosis with nuclear chromatin condensation, fragmentation and cell shrinkage was found by TEM. SP method showed that Celeib decreased COX-2 expression (17.48±0.34 vs 12.82±0.51,t=13.20,P =0.00). The sensitivity ratio(D0) was 1.15. FCM showed that the percentage of cells in G2/M phase was significanty more in R and D+R groups than in Ci and Cd groups (68.00±1.65,54.27±5.74,17.60±0.80,14.86±1.23, t=47.70,P=0.000; t=11.63, P=0.000), and also significantly different between R group and D + R group (t=3.99, P= 0.020). The apoptosis rate was higher in R and D + R groups than Ci and Cd groups(4.83±0.97,9.50± 1.35,1.33±0.86 and 2.28±0.42,t=4.67,P=0.010;t=8.81, P=0.000), D + R group than R group(t =4.85,P=0.010). Conclusions Celecoxib can markedly inhibit the growth and induce apoptosis in CNE-2 cells,which may depend on COX-2 pathway. Celeeoxib potently enhances the radiosensitivity of CNE-2 cells,which may due to the repair inhibit of radiation-induced DNA damage, inhibit of cell proliferation,and enhancement of cell apoptosis after irradiation.

Objective To observe how brain death affects the hepatic morphology and function of pigs and explore the roles of NF-κB. Methods Under general anaesthesia twelve healthy pigs were allocated randomly to two groups:control group(6 pigs),with non-inflacted Foley balloon catheter placed in the cerebral ventricle for 24 h,and brain death group,6 pigs,with estabhshment of brain death for 24 h.The serum and hepatic tissues in the same locus were taken at 6 h,12 h,and 24 h after the initial conformation of brain death.AST and ALT were determined by automatic biochemistry analyzer.IL-1βwas determined by ELISA.The NF-κB mRNA was determined by Real-time PCR and the NF-κB p65 by immunohistochemistry. Results The AST,ALT,IL-1β in serum,the NF-κB mRNA and the NF-κB p65 in hepatic tissues in brain death group were higher than those in control group and they all increased with the time(P<0.05).In brain death group,hepatocytes were edematous lightly after 12 hours,and the swelling progressively deteriorated after 24 hours,but there were no necrosis. Conclusion The activated NF-κB by brain death promoted the synthesis and release of inflammatory mediators,resulting in the hepatic dysfunction.

Objective To investigate micro-inflammatory state and protein-energy wasting (PEW) states in maintenance peritoneal dialysis(MPD) patients,then analysis of the correlation between them.Methods Ninty-six cases of MPD patients in this Hospital were selected from March 2012 to September 2015.The status of nutrition were assessed by Quantitative Subjective and global Assessment(SGA),malnutrition-inflammation score(MIS) and albumin(Alb),micro-inflammatory state was assessed by enzyme-linked immunoassay(ELISA) method serum hypersensitive c-reactive protein (hs-CRP),tumor necrosis factor-α(TNF-α),interleukin-6(IL-6).At the same time,various serological markers like serum Alb,serum total protein(TP),serum prealbumin(PA),hemoglobin(Hb),transferrin(TF),serum creatinine(Scr),urea nitrogen(BUN),cholesterol(Teh) were measured.Results The incidence of PEW in MPD patients was 36.50％,among which 62.86 ％ of them were over 65 years old,57.10％ were over 2 years of dialysis time and 40.00％ with diabetic nephropathy.MPD patients with hs-CRP＞5 mg/L accounted for 58.33％,of which over 65 year old accounted for 42.86％,MPD age longer than 2 years accounted for 60.71％,32.14％ of them with diabetic nephropathy.The proportion of diabetic nephropathy,average age,dialysis duration time,hs-CRP,TNF-α and IL-6 in PEW group were higher than non-PEW group(P＜0.05);BM,TP,Alb,PA,Hb,TCh,MAC and MAMC were lower ban non-PEW group(P＜0.05).Compared with the hs-CRP≤5 mg/L group,average age,the time of dialysis duration,TNF-α,IL-6 were higher and TP,Alb,PA,TF,Hb,the proportion of Kt/V≥1.72 were lower in the hs-CRP＞5 mg/L group.After the correction of age,sex,dialysis ages,it was found that the level of hs-CRP in MPD patients was negatively correlated with the level of Alb,PA,TF,Tch,Scr,TG;The level of IL-6 was negatively correlated with the levels of Alb,PA,TF,Tch,TG.The level of TNF-α in MPD patients showed different degrees of negative correlation with the leves of Alb,PA,TF,TG,Tch(all P＜0.05).Multivariate analysis showed that elderly,the time of dialysis duration,the microinflammatory state,and hypoalbuminemia were the independent risk factors of PEW.Conclusion PEW and micro-inflammatory state are very common in PHD patients.Patients with longer duration of dialysis,elderly or associated with diabetic nephropathy are more likely to suffer PEW and micro-inflammatory.Elderly,the time of dialysis duration,microinflammatory state,hypoalbuminemia are the independent risk factors of PEW.

Objective To measure the nursing workload in ICU,and provide the basis for the scientific distribution of human resources and reasonable scheduling in ICU.Methods ICU patients of the First and Third People's Hospital of Zigong from July to September,2011 were selected.The TISS-28 and NAS were applied to measure the nursing workload in the ICU.The number of staff nurses on each day and on each shift was recorded.Results The total nursing workload and the per capita nursing workload for each day in a week showed no significant difference,and the per capita nursing workload of different shifts and the needs of nurse-patient ratio of different patients in ICU were significantly different.Conclusions The allocation of nursing staff should be balance for each day in a week and different shifts for each day.The configuration of the nursing human resources should be based on actual nursing workload,no only considering the species of the diseases,the disease condition,self-care ability and cooperation of the patients,but also the proficiency of nurse skills.

Soft environment of opening laboratory in key subject will deeply affect the construction and development of disciplines.Developing and innovative academic team;scientific and rigorous running mechanism,healthy and concordant academic atmosphere,active and positive behavior mode,all of the above are the main factors of soft environment of laboratory.From now on,we need try our best to improve human rights,design more efficient aims,constitute impellingly guarantee system and fine mechanism of consistent of regulations and environment,apply modern managing instrument scientificly and improve managing efficiency.

Objective To investigate the relationship between the levels of Thl and Th2 cells and their cytokines IFN-γ,TNF-αt,IL-2,IL-4,IL-5 and IL-13 in peripheral blood of patients with acute pancreatitis and the clinical severity of acute pancreatitis,and to evaluate the diagnostic value and significance for the severity of acute pancreatitis.Methods This study enrolled 72 patients with acute pancreatitis and 30 healthy controls admitted to the Emergency Surgery Department of the First Affiliated Hospital of Zengzhou University from January 2015 to June 2017.The patients were divided into the mild MAP group,moderate MSAP group,and severe SAP group according to the 2012 Atlanta classification criteria.Whole blood and serum samples were taken on the day of admission for all confirmed cases,the number of Th1 and Th2 cells was detected by flow cytometry,the expression levels of IFN-γ,TNF-α,IL-2,IL-4,IL-5 and IL-13 were determined by ELISA.The total length of hospital stay and length of hospital stay in ICU of each group were recorded.The number of Th 1 and Th2 cells and the expression of related cytokines in patients with mild,moderate and severe acute pancreatitis were compared by one-way analysis of variance.Pearson linear correlation was used to analyze the correlation between Th1and Th2 cytokines and the length of hospital stay and the length of hospital stay in ICU.A P＜0.05 was considered statistically significant.Results Compared with the healthy controls,the number of Th1 and Th2 cells,the levels of Th1 cytokine IFN-γ,TNF-α IL-2 and Th2 cytokine IL-4 were significantly increased in acute pancreatitis.Th1 cells in the SAP group were significantly higher than those in the MAP and MSAP groups (F=11.137,P＜0.01),while Th2 cells in the SAP group were significantly lower than those in the MAP and MSAP groups (F=9.493,P＜0.01),and there was no significant difference between the MAP group and MSAP group.The expression level of IFN-γ was higher in the SAP and MSAP groups than that in the MAP group (F=8.605,P=0.001).The expression level of TNF-α was higher in the SAP group than those in the MSAP and MAP groups (F=1 1.847,P＜0.01),and the IL-4 expression level was significantly lower in the SAP group than those in the MAP and MSAP groups (F=8.042,P=0.001).The expression level of IFN-γ was positively correlated with the length of hospital stay and the length of hospital stay in ICU (r=0.569,P=0.014;r=0.538,P=0.021).The expression level of TNF-α was positively correlated with the length of hospital stay (r=0.475,P=0.046).The expression level of IL-4 was negatively correlated with the length of hospital stay and the length of hospital stay in ICU (r==0.577,P=0.012;r=-0.657,P=0.003).Conclusions Th1 and Th2 cells and their associated inflammatory factors IFN-γ,TNF-α,IL-2 and IL-4 are elevated in patients with acute pancreatitis.The increase in the number of Th1 proinflammatory cytokines IFN-γ,TNF-α and Th1 cells tends to aggravate the development of acute pancreatitis,and the increase in the number of Th2 cells and the anti-inflammatory cytokine IL-4 are associated with a better prognosis.