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.

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 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.

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 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 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 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.

Background and Objectives: Anticoagulation clinics are widely used for anticoagulation management in many countries, but have only recently began to gain acceptance in Taiwan. Our service model is a physician-managed outpatient clinic collaborating with clinical pharmacist and nurse. This study aimed to evaluate the adequacy of anticoagulation and rates of warfarin-related complications before and after referral to our collaborative anticoagulation clinic (CAC). Methods: Stroke patients taking warfarin from the neurology department were identifi ed and referred to the CAC during the 12-month period from February 2009 to January 2010. Quality markers include percentage of international normalized ratio (INR) values in the therapeutic range, frequency of INR monitoring, and frequency of follow-up visits and the mean interval of next INR monitoring after non-therapeutic INRs were compared one year before and after management in the CAC. Using studied patients as self-control, they were included in the analysis if patients had at least 3 months follow-up or 3 INR values both before and after referral. Results: A total of 44 stroke patients were included: mean age of 75.0 ± 9.7 years, with a CHADS2 score of 3.71 ± 0.69. The adequacy of anticoagulation was signifi cantly greater during CAC care compared with the period before referral; the percentage of INR within expanded therapeutic range was 60.9% versus 53.7%, respectively (p=0.049). Reduction in sub-therapeutic INR values from 31.8% to 24.2% (p=0.023) contributed mostly to the improved quality of care. The time interval of next INR monitoring after non-therapeutic INRs ( 4.0 or 1.5) was also signifi cantly shorter. However, there was no signifi cant difference in the rates of thromboembolic and hemorrhagic events which may be attributed to a small sample size. Conclusion: Based on results of our study, a CAC may be the optimal structure for anticoagulation management service in the future.

Objective:To study the expression of P53, Bax, Bcl-2 proteins and the role of cell apoptosis in the formation and development of acute radiation-induced skin ulcers.Methods:A rat model which was locally irradiated with 60 Co γ-rays was used, and the pathological changes were observed for 40 days. Immunohistochemistry and TUNEL assay were performed which enabled the detection of P53, Bax, Bcl-2 and cell apoptosis during the formation and development of radiation skin ulcers.Results: Skin ulcers were found on day 14 after irradiation, and enlarged and deepened gradually during the observation period. P53 was over expressed during days 11 to 40 after irradiation and was localized in vascular endotheliocytes and smooth muscle cells. Bax was moderately positive during days 14 to 21 and weakly positive during days 28 to 35, and was localized in vascular endotheliocytes, some fibroblasts and proliferating keratinocytes. Bcl-2 was weakly positive during days 1 to 11 after irradiation, and was located in keratinocytes, hair follicular cells and some vascular endotheliocytes. Bcl-2 was negative during days 11 to 40.The rate of cell apoptosis, especially of vascular endotheliocytes,wash igher than that in the early process of normal wound healing. Conclusions:After irradiation,the increased expression of the apoptosis-inducing protein P53, Bax and the decreased expression of the apoptosis-inhibiting protein Bcl-2 might be associated with the high rate of apoptotic events, and play important roles in the formation and development of radiation skin ulcers.