Objective To quantify the mRNA of 5-lipoxygenase (5-LO) in the rat peritoneal macrophages cultured in vitro by capillary electrophoresis (CE). Methods The rat peritoneal macrophages were isolated and cultured for indicated time. The mRNA of 5-LO was detected by RT-PCR, and the products of RT-PCR were quantified by CE. Results The DNA fragments in the 100 bp DNA marker and the products of the RT-PCR were separated successfully by CE with the sieving buffer containing 1.8% hydroxypropylmethylcellulose (HPMC). It proved that there were no significant changes on the expressions of 5-LO mRNA when the cells were cultured for 72 h quantified by CE. The mRNA of 5-LO significantly decreased by almost 80% by CE with the cells cultured for 120 h in vitro.Conclusions The products of RT-PCR could be separated and quantified by CE directly.The 5-LO mRNA could express normally in the rat peritoneal macrophages for 72 h in vitro.

Objective To investigate the anti-inflammatory effect of proanthocyanidins and its mechanisms.Methods Inflammation models such as dimethylbenzene-indueed ear swelling and carrageenan-induced hind paw edema in mice and rats were prepared.The contents of PGE2 in exudate from edema paws of rats were measured by ultraviolet spectrophotography and the protein expression of COX-2 in edema paws of rats by Western-blot and immunohistoehemistry(IHC)assay.Results Pro-anthocyanidins remarkably inhibited the ear edema induced by dimethylbenzene in mice at the dose of10 mg/kg and 20 mg/kg;paw edema of rats induced with carrageenan was significantly inhibited byproanthocyanidins 5,20 mg/kg ip from 2 to 5 h;proanthoeyanidins 5,20 mg/kg ip reduced the contents of PGE2 in exudate from edema paws of rats induced by carrageenan;proanthocyanidins 5,20 mg/kg ip inhibited the protein expression of COX-2.Conclusion Proanthocyanidins has an anti-inflammatory effect in vivo which may be related to inhibition of protein expression of COX-2 and downregutation of PGE2 biosynthesis.

Objective To evaluate the diagnostic accuracy of the combination of endorectal ultrasonography and serum CEA in preoperative diagnosis of rectal wall invasion (T staging) and nodal involvement (N staging) of rectal carcinoma.Methods We retrospectively analyzed clinical records of 310 patients with rectal carcinoma who underwent endorectal ultrasonography and serum CEA evaluation in Shanxi Province Tumor hospital from January 2007 to January 2010.The positive standard of CEA is more than 5 μg/L.The endorectal ultrasonography staging with postoperative pathological staging,and calculated the overall accuracy of T staging and N staging based on TRUS alone or on TRUS combined with serum CEA level were compared.Results The difference in serum CEA level was statistically significant from T1 to T4 (P ＜ 0.05).The accuracy rate of preoperative T staging of rectal carcinoma by TRUS alone was 71％ (219/310) and was 82 ％ (254/310) with TRUS combined with serum CEA level,showing significant statistical difference (x2 =10.92,P ＜ 0.01).The accuracy rate of preoperative N staging of rectal carcinoma was 69 ％ (211/308)with TRUS alone and was 77 ％ (238/308) with TRUS combined with serum CEA level,the difference of which was statistically significant (x2 =5.00,P ＜ 0.05).Conclusion Serum CEA level increases with an increasing pathological stage of rectal cancer.The combination of TRUS and serum CEA improves the accuracy of preoperative staging of rectal cancer.

Objective: In order to detect the effect of bcl-xs on camptothecin-induced apoptosis in human nasopharyngeal carcinoma CNE-2Z cells in vitro.Methods:bcl-xs gene-bearing mammalian expression vector(pcDNA3xs)was transfected into CNE-2Z cells using LipofectAmine.The expression of bcl-xs was determined with western blot.Cells which were transfected with native pcDNA3 vector were used as control.Apoptotic cells were detected with flow cytometry after exposure to camptothecin for 24h.Results:Cell clone(CNE-2Zxs)with stable expression of bcl-xs was obtained as confirmed with western blot.Results from flow cytometry analysis showed a significant increase of apoptotic cells in CNE-2Zxs as compared with CNE-2Zneo after treatment with the same dose of camptothecin.Conclusion:Exogenous bcl-xs expression sensitized nasopharyngeal carcinoma CNE-2Z cells to camptothecin-induced apoptosis.

Objective:To investigate the expression of microRNA-324-5p (miR-324-5p) and transcription factor forkhead box C1 (FOXC1) in glioma and their relationship with the prognosis of patients.Methods:From March 2012 to March 2015, a total of 72 cases of glioma tissues were collected from glioma patients who were admitted to Chongqing Hygeia Tumor Hospital and the People's Hospital of Nanchuan in Chongqing, and 28 cases of normal human brain tissues resected in craniocerebral surgery were also collected. The expressions of miR-324-5p and FOXC1 mRNA were detected by real-time fluorescent quantitative polymerase chain reaction (qRT-PCR), and the expression of FOXC1 protein was detected by immunohistochemistry. Pearson method was used to analyze the correlation between the expressions of miR-324-5p and FOXC1 in glioma tissues; Kaplan-Meier method was used to analyze the survival of patients with glioma; Cox regression analysis was used to analyze the risk factors affecting the prognosis of patients with glioma.Results:FOXC1 protein was mainly located in the cytoplasm of glioma, and its positive expression rate in glioma tissues was 81.94% (59/72), which was significantly higher than that in normal brain tissues [17.86% (5/28)], and the difference was statistically significant ( χ2 = 35.938, P<0.01). Compared with normal brain tissues, the expression of miR-324-5p was down-regulated in glioma tissues (0.62±0.19 vs. 0.98±0.02, t = 9.974, P < 0.05), and the expression of FOXC1 mRNA was up-regulated (1.41±0.29 vs. 0.99±0.02, t = 7.633, P < 0.05). The expressions of miR-324-5p and FOXC1 protein were correlated with the number of primary lesions, differentiation degree, TNM stage and lymph node metastasis of glioma (all P<0.05). Pearson analysis showed that the expressions of miR-324-5p and FOXC1 mRNA were negatively correlated ( r = -0.550, P<0.01). The 5-year overall survival rate of patients in miR-324-5p high-expression group was significantly higher than that of patients in miR-324-5p low-expression group (45.71% vs. 24.33%, χ2 = 6.531, P = 0.011), and the 5-year overall survival rate of patients in FOXC1 protein high-expression group was significantly lower than that of patients in FOXC1 protein low-expression group (30.41% vs. 42.34%, χ2 = 3.631, P = 0.047). Multivariate Cox regression analysis showed that low differentiation, TNM stage Ⅲ-Ⅳ, lymph node metastasis, low expression of miR-324-5p and high expression of FOXC1 protein were independent risk factors for prognosis of glioma patients (all P < 0.05). Conclusions:The expression of miR-324-5p is low and the expression of FOXC1 is high in glioma. They may be involved in the regulation of tumor differentiation and metastasis, and related to the poor prognosis of patients. They may be potential therapeutic targets for glioma.

Cardiopulmonary exercise test and electrocardiogram exercise test were performed in 68 patients with suspected coronary artery disease,the diagnostic value of two tests was compared with coronary angiography results as the gold standard.The results show that the sensitivity,specificity,positive predictive value and negative predictive value of electrocardiogram exercise test for coronary heart disease were 51.28％,68.97％,68.97％ and 51.28％ respectively; those for ratio of O2 pulse peak in cardiopulmonary exercise test were 51.28％,75.86％,74.07％ and 53.66％ respectively.It suggests that the results of cardiopulmonary exercise test may have the same value as electrocardiogram exercise test in diagnosis of coronary artery disease.

OBJECTIVETo observe the changes of tumor necrosis factor alpha (TNF alpha), interleukin 8 (IL-8) in liver ischemia/reperfusion injury and the protective effect of hydrophilic Salvia monomer MP-1 on them.

METHODSHypothermic hypoxia reoxygenation model of human liver sinusoidal endothelial cell line and ischemia/reperfusion model of isolated rat liver were used. TNF alpha and IL-8 were measured with ELISA kits. Cell injury was excluded by trypan blue stain, sinusoidal endothelial cell function was assessed by hyaluronic acid uptake rate through RIA. Liver function was assayed by alanine transaminase (ALT), aspartate aminotransferase (AST) and lactate dehydrogenase (LDH) release as well as output of bile flow.

RESULTSDuring hypoxia reoxygenation, sinusoidal endothelial cell injury, TNF alpha and IL-8 increased significantly in time-dependent manner, while sinusoidal cell function decreased. Cell injury was positively correlated to the released amount of TNF alpha (r = 0.949, P < 0.05) and IL-8 (r = 0.892, P < 0.05), respectively, the mortality could be reduced within 6 hrs by adding anti-TNF alpha monoclonal antibody and increased by treating with recombinant human TNF alpha. Function of isolated rat liver lowered alone the increasing of low temperature ischemia/reperfusion time. MP-1 could markedly lower the mortality of endothelial cells and TNF alpha and IL-8 release, it also could alleviate ischemia/reperfusion injury to isolated rat liver.

CONCLUSIONTNF alpha participated in liver ischemia/reperfusion injury directly, and MP-1 might alleviate the injury through inhibiting TNF alpha and IL-8.

Animals ; Cell Hypoxia ; Cells, Cultured ; Drugs, Chinese Herbal ; pharmacology ; Endothelium ; cytology ; metabolism ; In Vitro Techniques ; Interleukin-8 ; metabolism ; Liver ; blood supply ; cytology ; Male ; Phytotherapy ; Protective Agents ; pharmacology ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; metabolism ; Salvia miltiorrhiza ; chemistry ; Tumor Necrosis Factor-alpha ; metabolism

Objective To assess value of serum level of ischemia modified albumin (IMA) in diagnosis for myocardial ischemia of coronary artery disease (CAD). Methods Seventy-two patients with clinically suspected myocardial ischemia of CAD admitted to The First People's Hospital of Hangzhou during November 2009 to May 2010 ready for undergoing coronary angiography, the gold standard for diagnosis of CAD, were randomly selected for the study. The patients were divided into CAD and non-CAD groups based on their coronary angiography. Serum level of IMA was determined with cobalt-albumin binding ( ACB) assay before coronary angiography, which served as diagnostic standard for CAD. Logistic regression analysis method was used to evaluate varied levels of IMA with area under the receiver operating characteristic curve (AUCROC) in diagnosis for myocardial ischemia of CAD. Results Mean level of IMA was (97 ±24) U/ml and (81 ±15) U/ml for CAD group (n =51) and non-CAD group (n =21), respectively. Sensitivity and specificity of a cut-off value of IMA 83.69 U/ml in diagnosis for myocardial ischemia of CAD was 80 percent and 57 percent, respectively, with a predictive value of a positive test 82 percent and that of a negative test 55 percent, respectively, from AUCROC. Logistic regression analysis demonstrated that both hypertension (P=0. 022, 6 = 1.421, OR=4. 141) and level of IMA (P=0.003, b= 1.780, OR=5.928) were independent predictors for CAD. Conclusions Sensitivity, specificity and predictive value of a positive test of the level of IMA are relatively high in diagnosis for myocardial ischemia of CAD, which is an independent predictor of it.

Objective To investigate the factors associated with postoperative complications after laparoscopic-assisted radical surgery in rectal cancer.Methods The clinical data of 310 patients with rectal cancer performed by laparoscopic-assisted radical resection from November 2010 to August 2013 were analyzed retrospectively.The differences between patients with and without postoperative complications were compared.All the data were analyzed by the t test,chi-square test or Logistic regression analysis.Results Among the 310 patients,postoperative complication occurred in 80 patients.On univariate analysis,postoperative complication was associate with gender,age,body mass index,preoperative comorbidity,diameter and location of tumor,TNM staging,operative time and surgeon experience (all P ＜ 0.05).Logistic regression analysis revealed that gender,body mass index,preoperative comorbidity,location of tumor,TNM staging and surgeon experience were independent risk factors for postoperative complications (all P ＜ 0.05).Follow-up was available in 260 patients,with a median follow-up of 18 months (3-30 months).Differences in survival rates between patients with and without postoperative complications were no statistical significance (x2 =1.201,P =0.273).Conclusions Gender,body mass index,preoperative comorbidity,location of tumor,TNM staging and surgeon experience are independent risk factors for postoperative complications in laparoscopic-assisted radical surgery for rectal cancer.The short and medium-term survival time between patients with and without postoperative complications are similar.

Objective To explore effects of Naikan cognitive therapy on improving clinical symptoms in patients with convalescent schizophrenia. Methods The 69 convalescent schizophrenic patients with convalescent clinical state were consecutively recruited. All the patients were divided into Naikan cognitive therapy ( NCT) group an control group at random and were pretreated with antipsychotic agent therapy. In NCT group,the patients received NCT for successive 7 days. In control group,the patients only received antipsychotic agent therapy. Pre-and post-treatment positive and negative syndrome scale( PANSS) , Nurses'observation scale for inpatient evaluation (NOSIE) were administered to all subjects. Results ① A significant decrease occurred in PANSS total score, negative symptom score, positive symptom score, compound scale score, general psychopathology score, reaction retardation score and paranoid score in NCT group ( t = 2. 672～7. 370, P < 0. 05). In the post-treatment, PANSS total score, negative symptom score, positive symptom score, compound scale score, reaction retardation score and thought disorder score were significantly lower in NCT group than those in control group ( t ' = 2. 696, P = 0. 009; t = 5. 186, P=0.000; t = 3.757, P = 0.001; t = 2.634,P = 0.011; t ' =2.376, P = 0.021). ②A significant decrease occurred in NOSIE total negative score( 10.43 ± 9. 24 vs 13. 87 ± 8. 03, t = 3. 463 , P = 0. 002) , irritation score(3. 13 ±0.43 vs8.53 ±4.98, t = 6. 139, P=0.000) and retardation score(1.07 ± 1.64 vs 2. 20 ±2.85, t = 2.067, P = 0.048) in NCT group. Conclusion NCT can possibly improve part clinical symptoms of patients with convalescent schizophrenia to a certain extent,especially negative symptom,but need to further prove the effect of NaiKan cognitive therapy.