ObjectiveTo observe the apoptosis of breast cancer cells induced by small direct-current electric field (EF) and its related possible mechanism in expression of p21 and p16 genes.MethodsSmall direct-current EF was applied in human breast cancer cell line MCF7 in vitro and the effect of EF on apoptosis and expression of p21 and p16 important genes of breast cancer cells was observed.ResultsThe small direct-current EF had an obvious apoptosis inducing effect on breast cancer cells. A lot of cancer cells were sloughed from culture dish by 200 mV/mm of the EF strength. The number of the apoptotic cancer cells was gradually significantly increased and mRNA expressions of p21 and p16 genes were raised following the EF strength being up (P<0.01).ConclusionThe small direct-current EF has an obvious apoptosis inducing effect on breast cancer cells. It may be related with the expression of p21 and p16 genes.

ObjectiveTo research the apoptosis of breast cancer cells by application of small direct-current electric field(EF) and its related possible mechanism.MethodsSmall direct-current EFs were applied in human breast cancer cell line MCF7 in vitro to observe the effects on apoptosis of cancer cells by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL)and to detect the mRNA expression of p53, Rb and E2F1 genes by reverse transcriptase-polymerase chain reaction (RT-PCR).ResultsA lot of cancer cells were observed slaughed from culture dish by 200mV/mm of the field strength and the number of the apoptotic cancer cells was gradually significantly increased with the application of EFs(P<0.01). mRNA expression level for p53 and Rb genes was elevated but that for E2F1 gene was decreased overtly in cancer cells exposed to EFs.ConclusionApoptosis of breast cancer cells can be induced by small direct-current EFs and the change for expression of p53, Rb and E2F1 genes may be involved in the mechanism.

AIM:To construct the adenovirus vector with adiponectin (Acrp30) siRNA, and to observe its effect on the Acrp30 expression and glucose transport in 3T3-L1 adipocytes. METHODS: Mouse Acrp30 siRNA fragment was designed, synthesized and cloned into the adenovirus vector. 3T3-L1 cells were infected with the two recombinant adenoviruses, respectively. The mRNA expression and protein levels of Acrp30 in these cells were evaluated by RT-PCR and ELISA. Glucose transport was measured by 2-Deoxy-[3H]-D-glucose incorporation method. RESULTS: The recombinant adenoviruses were successfully constructed. They remarkably downregulated the expression of Acrp30 at both mRNA and protein levels in 3T3-L1 cells, and decreased the glucose transport in 3T3-L1 adipocytes (P

Objective To study the content changes of wilforlide A in Xinfeng capsules before and after compatibility.Methods The content of wilforlide A was determined in Xinfeng capsules ester content and in Tripterygium wilfordii Hook.f.single solution by TLCS method.Developing solvents:chloroform and methanol (85:1),scanning wavelength:230 nm.Results Wilforlide A showed a good linear relationship in 1.700-5.100p g,the regression equation is Y =1 118.766 + 3 253.751 X (r =0.999).The average recovery of the wilforlide A in Xinfeng capsules was 102.84％,and RSD was 3.64％.The average recovery of the wilforlide A in the Tripterygium wilfordii Hook.f.single decoction was 98.14％,and RSD was 2.21％,compatibility levels before and after the change (compatibility with 0.179mg/g,0.136mg/g after compatibility).Conclusion The established TLCS method was simple,specific and accurately quantitative.The comparative study found that the compatibility of Xinfeng capsules Tripterygium reduced the content of wilforlide A,which may be related to decreasing toxicity.

Objective To assess the clinical significance of antibodies against cyclic citrullinated peptide(anti - CCP) in a cohort of patients with juvenile idiopathic arthritis (JIA). Methods The anti - CCP was tested by an enzyme linked immunosorbent assay (ELISA) in serum samples from 22 patients with JIA(9 boys, 13 girls), with a mean age of 9.1 years(range 1.1 - 16). As control groups, anti - CCP were also tested in sera of 20 healthy children, and 30 adult patients with rheumatoid arthritis(RA). Results Positive anti - CCP were found in sera of 2 patients with JIA(9.1 %), both with lower titre comparing with those of adult patients with anti - CCP positive. In the control groups, none of healthy children was positive for anti - CCP, but 16/30(53. 3%) adults with RA showed anti - CCP positivity. Conclusion Anti - CCP can be detected in children with JIA, but are less frequently present than in adults with RA, so anti - CCP has limited diagnostic value to JIA.

Objective To investigate the clinical features and prognosis of rectal neuroendocrine neoplasms (NENs) in different pathological grades.Methods The clinical data of 183 patients with rectal NENs who were admitted to the PLA General Hospital from January 2001 to April 2012 were retrospectively analyzed.All the clinical and pathological data of the patients who received endoscopy and (or) surgical resection were retrieved from the work station and the database of the endoscopic center.Based on the 2010 WHO pathology classification of digestive tumors,the pathological data were ranked according to the mitotic count.The prognosis of the patients was learned by re-examination or phone call.The follow-up ended till July 2014 or at the death of patients.Data were analyzed using the chi-square test.Results A total of 183 patients were enrolled in this study including 120 males and 63 females.The median age of the patients was 48 years (range,14-83 years).Seventy-four patients had the symptom of hemafecia,9 patients had abdominal pain and change in bowel habit,and 70 patients were diagnosed by body examination.Other symptoms included increased level of tumor markers and abdominal distension.Carcinoid syndrome was not detected in all the patients.Fourteen patients were complicated with polyp of intestine,5 with tubular adenoma,3 with colorectal adenocarcinoma and 1 with small cell lung cancer.The diameters of the tumor under 1 cm were detected in 162 patients,the diameters of the tumors ranged between 1 cm and 2 cm in 14 patients,and the diameters of the tumors above 2 cm in 7 patients.The mean distance between the tumor and the anus was (5-± 3)cm.Of the 183 patients,130 received endoscopic treatment,43 received surgical treatment and 10 received clamping because the tumor was misdiagnosed as polyps.There were 158 patients in grade1 (154 in stage Ⅰ,1 in stage Ⅱ,1 in stage Ⅲ and 2 in stage Ⅳ),21 in grade2 (13 in stage Ⅰ,3in stage Ⅱ,3 in stage Ⅲ and 2 in stage Ⅳ),4 in grade 3 (1 in stage Ⅰ,1 in stage Ⅲ and 2 in stage Ⅳ).Six patients had liver metastasis and 9 had lymph node metastasis.Fourteen patients died (4 in grade 1,6 in grade 2 and 4 in grade 3).The 5-year survival rate of patients was 92.35％ (169/183).There were significant differences in the gender,tumor diameter,tumor staging,lymph node metastasis distal metastasis and 5-year survival rate among patients with rectal NENs in different pathological grades (x2=60.949,71.587,32.135,55.486,56.512,P ＜ 0.05).Conclusions Rectal NENs lacks the specific clinical manifestation and is more likely to happen in males,and it often locates at the middle-lower rectum.Most of the rectal NENs belongs to stage Ⅰ and grade 1 and is less than 1 cm in size.The prognosis of patients with rectal NENs in different pathological grades is different.The 2010 WHO pathology classification of digestive tumors is useful to asses the prognosis of rectal NENs.Different grades of rectal NENs could be taken into account when designing the treatment plan.

Objective To study the related prognostic factors influencing rectal neuroendocrine tumor prognosis.Methods We retrospectively reviewed tumor clinical and pathology material of 183 cases with rectal neuroendocrine in our hospital during recent 10 years and reclassified them according to the 2010 WHO pathology classification.Single factor and multivariate analysis were performed to find related prognostic factors.Results Single factor analysis showed that tumor size (＜ 1 cm,1-2 cm,＞2 cm,98.77％ vs 78.57％ vs 28.57％,x2 =71.4793,P =0.0001),pathological grading (G1,G2,G3,98.73％ vs.76.19％ vs 25.00％,x2 =56.5121,P =0.0001) and the stage (stage Ⅰ,Ⅱ,Ⅲ,Ⅳ period,99.40％ vs 50.00％ vs 60.00％ vs 16.67％,x2 =105.0383,P =0.0001) among the three variables survival difference was statistically significant.Multivariate analysis (Cox regression) indicated that the tumor stage were independent factors affecting the prognosis,regression coefficient was 1.277 (B),Wald value of 28.145,Exp (B) a value of 3.586,95％ CI:2.237-5.747.Conclusion Tumor staging is an independent risk factor of survival,without considering other factors,different tumor size and pathologic grade also have different prognosis.Clinicians in making treatment plan should fully consider the above factors before choosing the appropriate follow-up plan.

Objective To construct the bait expression plasmid pGBKT7-GR of glucocorticoid receptor(GR)binding domain.Methods The fragments of GR binding domain was amplified by RT-PCR,and then was cloned into pMD18-T.After being verified by sequencing,it was subcloned into the bait expression vector pGBKT7.Then the bait vector pGBKT7-GR was transformed into AH109 yeast cells and the expression of the bait protein was analyzed by Western blot.Toxicity and self-activation of the bait protein were detected.Results GR binding domain was amplified and cloned into pMD18-T and pGBKT7 successfully.The bait vector was transformed into AH109 yeast cells successfully,without toxicity or self-activation.The expression of the bait protein was confirmed by Western blot.Conclusion The successful construction of bait expression vector of glucocorticoid receptor binding domain lays the foundation for constructing small molecule ligand yeast three-hybrid system.

Objective To investigate the risk factors and early diagnosis of the severe thrombocytopenia complicating transcatheter ccclusion of patent ductus arteriosus ( PDA ) . Methods Between February, 2011 and May, 2015, 80 patients with patent ductus arteriosus underwent percutaneous intervention occlusion were studied. Results Average age were ( 17. 5 ± 17. 1 ) years, 63 were females (78. 8%), mean weight were (35. 6 ± 20. 2)kg (from 6. 0 to 75. 0 kg), mean body surface area (BSA) were (1. 09 ±0. 44) m2(from 0. 32 to 1. 91 m2). A bolus of heparin calcium (80 U/kg) was administered by intravenous injection. The mean diameters of patent ductus arteriosus were 4 mm (from 2 to 18 mm), and the mean diameters of occluders were 12 mm (from 6 to 30 mm). 14 patients were found to have severe thrombocytopenia (PLT count﹤100 × 109/L). The reduction rate of platelet in 12 of 14 patients was more than 19%. The diameters of all occluders were equal to or more than 14 mm, the mean diameters of patent ductus arteriosus were 10 mm ( from 6 to 18 mm) and the mean diameters of occluders were 18 mm ( from 14 to 30 mm). All the 14 patients started to present progressive decrease in PLT count since the second day post procedure. Taking the selected occluder diameter greater than 14 mm as cut-off points in diagnosis of severe thrombocytopenia, the sensitivity was 100%, specificity was 68%, the positive predictive value was 40%, and the negative predictive value was 100%. Combined with the postprocedural second day complete blood count analysis and the platelet count decreased by 10% as cut-off points in diagnosis of severe thrombocytopenia patients, the sensitivity was 93%, specificity was 67%, the positive predictive value was 65%, the negative predictive value was 93% . If taking the platelet count decreased by 7% on second day as cut-off points in diagnosis of severe thrombocytopenia patients, the sensitivity was 100%, specificity was 57%, the positive predictive value was 61%, the negative predictive value was 100% . Logistic regression analysis discovered that risk factors of severe thrombocytopenia after PDA are procedural platelet count and occluder diameter. Conclusions The risk factors of severe thrombocytopenia complicating transcatheter ccclusion of patent ductus arteriosus were the procedural reduction of platelet count and big occluder diameter. Patients with PDA who were inplanted with occluders equal to or bigger than 14 mm should retest the numbers of platelet on the second day after procedure and retest on third day if the numbers reduce on the second day, which may help in the prediction of severe thrombocytopenia.

Objective:To observe influence of simvastatin on differentiation ,proliferation ,migration and adhesion of marrow-derived smooth muscle progenitor cells (SPCs) and screen coated eluting stent drugs of new generation . Methods :The mononuclear cells (MNCs) were isolated from rat marrow by density gradient centrifugation method , and then plated on fibronectin-coated culture dishes ,after culture 8d ,marrow-derived SPCs were identified by α-smooth muscle actin (α-SMA) immunofluorescent staining and counted under inverted fluorescence microscope .The MNCs and adhesion cells were treated with simvastatin (0.01~10 μmol/L) respectively for 8 d and 24h .SPCs pro-liferation ,migration and adhesion were observed by Tritium thymidine (3 H-TdR) intake method ,modified Boyden chamber assay and adhesion assay .Results:Compared with control group (no simvastatin intervention ) ,0.01μmol/L simvastatin significantly inhibited the MNCs differentiation towards SPCs [ (85 ± 4) vs .(79 ± 5)] ,proliferation [ (4070 ± 184) vs .(3833 ± 126)] ,migration [ (44 ± 3) vs .(39 ± 3)] and adhesion of SPCs [ (59 ± 5) vs .(52 ± 4)] , P<0.05 all ,and number of SPCs significantly reduced along with simvastatin concentration increased (P<0.01) . Conclusion:Simvastatin could inhibit the differentiation ,proliferation ,migration and adhesion of marrow-derived smooth muscle progenitor cells .