Objectives Esophageal cancer development and progression is driven by the accumulation of genetic changes. In this study, we used tissue microarray to survey the expression of p53, p16 and COX-2 in esophageal cancer progression and their clinical implication. Methods We tested p53, p16 and COX-2 proteins by immunohistochemistry and tissue microarray in 86 specimens from different stages of esophageal cancer and 40 specimens from adjacent non-cancer tissue. Results The expression of p53 and COX-2 was significantly higher in tumor tissue than that in non-tumor ones. As for the expression of pl6, no significant difference was found between tumor and adjacent tissue. An obvious relation was observed among p53, p16 and COX-2 expression that esophageal carcinogenesis was highly correlated with the positive expression of p53 or COX-2, however, no reciprocal relationship to neoplastic progression was recognized with p53, p16 and COX-2. Conclusion We observed that the tissue with the positive expression of p53 or COX-2 was more likely to develop esophageal cancer. Further work will verify the hypothesis that the expression level of p53 and COX-2 in biopsy specimen is applicable to predict early outset of esophageal cancer.

Objective To study on the relations between endometrial carcinoma and Infections of HSVⅡ.Methods Using to skill of PCR-SSCP to test 30 cases of endometrial carcinoma,others 15 cases of the hyperplastic endometrial and 15 cases of the normal of endometrial were infected HSVⅡ.Results Positive rates of endometrial carcinoma that was infected of HSVⅡ were 50%(15/30),Positive rates of the hyperplastic endometrial that was infected of HSVⅡ were 6.67%(1/15),15 cases of the normal of endometrials were not infected.Compared with the endometrial carcinoma and the hyperplastic endometrial and the normal of endometrial,P

ObjectiveTo compare the efficacies between conventional acupuncture plus acupoint application and conventional acupuncture plus TDP in treating dysmenorrhea due to blood stagnation led by cold.MethodSixty patients were randomized into two groups, 31 cases in the treatment group and 29 cases in the control group. The treatment group was intervened by conventional acupuncture plus acupoint application, while the control group was by conventional acupuncture plus TDP. Before and after the intervention, McGill Pain Questionnaire was adopted to compare the Pain Rating Index (PRI) score, Visual Analogue Scale (VAS) score, and Present Pain Index (PPI) score, and the therapeutic efficacy was also compared.ResultThe recovery rate was 58.1% in the treatment group, significantly higher than 24.1% in the control group (P<0.01); the total effective rate was 90.3% in the treatment group, significantly higher than 62.1% in the control group (P<0.01), indicating that the therapeutic efficacy of the treatment group was more significant than that of the control group. Before the intervention, there were no significant differences in comparing the PRI, VAS, and PPI scores between the two groups (P>0.05), suggesting the comparability. After the intervention, there were significant differences in comparing the PRI, VAS, and PPI scores between the two groups (P<0.05), and the PRI, VAS, and PPI scores were markedly decreased in the treatment group.ConclusionConventional acupuncture plus acupoint application can improve dysmenorrhea symptoms and reduce patient’s pain, and can produce a more significant efficacy compared with conventional acupuncture plus TDP.

Objective To investigate the response of cryopreserved ovarian tissue after autologous implantation in mouse stimulated with gonadotrophin.Methods Thirty six female mice were randomly divided into three groups,with 12 mice in each group.In group of fresh ovarian tissue,fresh ovarian tissue was implanted into kidney capsule of mice:in group of cryopreserved ovarian tissue,ovarian tissue was implanted into kidney capsule of mice after cryopreserved by vitrification for two weeks.We investigated the response of ovarian tissue two weeks later after autologous implantation stimulated with gonadotrophin.Immunohistochemistry staining method was used to observe the expression of follicle stimulating hormone receptor.Results Before and after stimularian with gonadotrophin,the mature follicle rate of group of fresh ovarian tissue was 2.3%and 4.2%.that of group of cryopreserved ovarian tissue was 2.3%and 4.0%,and that of group of control was 2.6%and 5.8%.Regarding the percentages of mature follicle.there were significant differences after stimulation with gonadotrophin(P＜0.05).After stimulating with gonadotrophin the percentages of mature follicle were the same in the fresh tissue group,cryopreserved tissue group and control group(P＞0.05).The integrated optical density of follicle stimulating hormone receptor of fresh ovarian tissue in antrofollicle and pre-antrofollicle were 9408±2777 and 4531±1903.that of cryopreserved ovarian tissue were 9175±3093 and 4808±1386.and that of the control ovarian tissue were 8838±2064and 5516±1136 respectively.There was no significant difference between any two groups(P＞0.05).Conclusion The follicle stimulating hormone receptor is preserved by cryopreservation and transplantation,small pieces of ovarian tissue response to gonadotropin stimulation is normal.

The bioactivity of a working reference standard was determined by replicate bioassays with calibration against a primary reference standard. In this study the number of bioassay replicates needed for calibration first was calculated theoretically, and if the mean value of the experimental bioassay replicates fell within the predefined bioactivity level the bioactivity of the working reference was defined as 100%. Our results showed that when the total intermediate precision of the bioassay method was at 11.66% and the predefined bioactivity level was set at 95%-105% with a confidence level of 95%, 21 bioassay replicates should be carried out for calibration. The average value of the 22 experimental bioassay replicates was 101.96%, so the bioactivity of the working reference standard was consistent with that of the primary reference standard at 100%. The results suggest that a strategy of first calculating the number of bioassay replicates needed for calibration and then determining whether the resulting experimental mean value is within the predefined bioactivity level will be of value to the biopharmaceutical industry.

Background and purpose: High expression of telomerase and telomere stability are two common features in tumor cell. hTERT is a catalytic subunit of telomerase, TRF2 is extremely important to maintain the length and stability of telomerase. This study was to construct the recombinant adenovirus mediated shRNA to hTERT and TRF2, and to investigate the inhibitory effects of the vector by solo-inhibiting and connect-inhibiting in the MCF-7 cells, in order to present a new approach to the gene therapy for breast cancer. Methods: rAd-hTERT and rAd-TRF2 were constructed and the expression of hTERT mRNA and TRF2 mRNA were tested by FQ-PCR 48 hours after transfecting in MCF-7 cells. Apoptosis was observed by flow cytometry 1 to 6 days after transfection. Results: ①At 48 hours after transfection, the results of FQ-PCR showed that compared to PBS group, the expression of hTERT in rAd-hTERT group was obviously decreased and the inhibition ratio was about 86%, but TRF2 had not been obviously inhibited (P>0.05);the expression of TRF2 in rAd-TRF2 group was obviously decreased and the inhibition ratio was about 80%, but hTERT had not been obviously inhibited (P>0.05);in rAd-hTERT/rAd-TRF2 group, the inhibition ratio of hTERT and TRF2 were about 88% and 85%. Comparing rAd-hTERT/rAd-TRF2 group with rAd-hTERT group and rAd-TRF2 group, there were no significant differences of inhibition ratio between hTERT gene and TRF2 gene(P>0.05). Otherwise, comparing rAd-HK group, rAd-blank group with PBS group, there were no significant differences of inhibition ratio between hTERT gene and TRF2 gene(P>0.05). ②The result of flow cytometry showed that apoptosis was induced at the first day after transfecting in rAd-hTERT group and rAd-TRF2 group, the most obvious apoptosis was in the 3rd to 5th days,at the peak in the 5th day, and decreased in the 6th day after transfection. The apoptosis ratio of rAd-hTERT group was 46.2%, rAd-TRF2 group was 43.5%. The apoptosis ratio of rAd-hTERT/rAd-TRF2 group was 46.2% at first day, 68.5% at the second day, the most obvious apoptosis was in the 3rd to 6th days and was 77.6% in the 6th days in rAd-hTERT/rAd-TRF2 group. There were significant differences in apoptosis ratio in solo-inhibiting and connect-inhibiting(P<0.05). In addition, comparing rAd-HK group, rAd-blank group with PBS group, there were no significant differences in apoptosis ratio(P>0.05). Conclusion: ①Target sequence of RNAi which aimed at hTERT gene and TRF2 gene was designed efficiently, and the RNAi expression vectors were seen in vivo study efficiently and specifically inhibited the correspond gene expression and promoted cell apoptosis in MCF-7 cells. ②rAd-hTERT vector and rAd-TRF2 vector have no synergistical effect and antagoinstical effect on inhibiting hTERT gene and TRF2 gene mRNA expressing in MCF-7, but there was synergistical effect in terms of the induction of apoptosis. So association-RNAi-technique targeting to the genes of telomere length and stability can effectively promote tumor cell into apoptosis and inhibit breast cancer cell growth. RNAi technique of connecting correlation genes is a more effective gene therapy strategy.

Objective To investigate the risk factors of gallbladder carcinoma,so as to provide theoretical base for the prevention of gallbladder carcinoma.Methods The clinical data of 153 patients with gallbladder carcinoma (gallbladder carcinoma group) who were admitted to the Peking Union Medical College Hospital from January 2000 to December 2010 were retrospectively analyzed. A total of 300 patients with cholecystolithiasis (cholecystolithiasis group) and 300 patients without gallbladder carcinoma or cholecystolithiasis (control group)were collected and matched at the ratio of 1∶2 to conduct the controlled study.Data were statistically analyzed by the Chi-square test and conditional Logistric regression.Results Univariate analysis showed significant difference in age,history of cholecystolithiasis,postmenopausal age,accumulated menstrual period,giving birth or not and number of birth between gallbladder carcinoma group and control group ( x2 =58.22,180.14,9.59,24.30,18.66,15.17,P ＜0.05).Age,history of cholecystolithiasis,accumulated menstrual period and number of birth were the independent risk factors of gallbladder carcinoma (x2 =55.76,180.95,24.30,8.54,P ＜ 0.05).The risk of having gallbladder carcinoma in patients who had a history of cholecystolithiasis was 34 times higher than those who did not have the history of cholecystolithiasis (OR =34.22).Late postmenopausal age (51 -55 years old),longer accumulated menstrual period ( ≥30 years),and the number of birth ( 3 times) were associated with higher risk of gallbladder carcinoma (OR =3.96,9.68,3.51 ). Age,course of cholecystolithiasis and accumulated menstrual period and number of birth were the risk factors of gallbladder carcinoma when comparing patients who have history of cholecystolithiasis in the gallbladder carcinoma group with those in the cholecystolithiasis group (x2 =70.66,16.66,11.59,4.69,P ＜ 0.05 ).Age,course of cholecystolithiasis and accumulated menstrual period were the independent risk factors of gallbladder carcinoma ( x2 =64.29,8.82,5.58,P ＜ 0.05).The risk of gallbladder carcinoma increased as the increase of age and course of cholecystolithiasis. The accumulated menstrual period ≥ 30 years was also a risk factor of gallbladder carcinoma. Conclusions Age,history of cholecystolithiasis,course of cholecystolithiasis,accumulated menstrual period and number of birth may be the risk factors of gallbladder carcinoma.For patients with age above 60 years and course of cholecystolithiasis above 3 years,cholecystectomy should be conducted to reduce the incidence of gallbladder carcinoma,and great importance should be attached to female patients with indications mentioned above.

Objective To analyze the clinicopathological features of gallbladder adenosquamous carcinoma.Methods The clinical data of 8 patients with gallbladder adenosquamous carcinoma who were admitted to the Peking Union Medical College Hospital from January 2000 to December 2010 were retrospectively analyzed.The clinicopathological features,diagnosis and treatment strategies were summarized.The survival curve was drawn by Kaplan-Meier method,and the survival of the patients was analyzed by Log-rank test. Results The clinical manifestation of the 8 patients was non-specific,and all the patients were misdiagnosed as with primary gallbladder cancer complicated with cholecystolithiasis or chronic cholecystitis.The levels of carcinoembryonic antigen and γ-glutamyl transferase of 3 patients were higher than the normal,the level of alkaline phosphatase of 2 patients was higher than the normal,the level of aspartate transaminase of 1 patient was higher than the normal,the level of CA242 of 3 patients was higher than the normal and the level of CA19-9 of 4 patients was higher than the normal.The levels of aspartate transaminase,total bilirubin and direct bilirubin were in the normal range. Different surgical approaches were selected according to different tumor positions.The mean diameter of the tumor was (5 ±3)cm. Lymph node metastasis was detected at the hepatic hilar,hepatoduodenal ligament and area behind pancreatic head. Postoperative pathological examination confirmed that all the patients were with gallbladder adenosquamous carcinoma. All patients were followed up till April 2011. Two patients survived with no discomfort,and the survival time was 2-3 months.Six patients died of tumor recurrence or metastasis,and their median survival time was 8 months (range,4-13 months). Conclusions The clinical manifestation of gallbladder adenosquamous carcinoma is non-specific, and definite diagnosis depends on the results of postoperative pathological examination.Surgical treatment is the only effective method in improving the prognosis of patients with gallbladder adenosquamous carcinoma,and the prognosis of patients is poor.

Objective To compare the various surgical procedures commonly used to treat ampullary neoplasms.Methods The clinical data of 238 patients with ampullary tumor who were admitted to the Peking Union Hospital from January 1995 to June 2012 were retrospectively analyzed.There were 51 patients with benign ampullary adenomas (34 patients treated with local tumor resection,17 patients treated with pancreaticoduodenectomy) and 187 patients with ampullary cancer (25 patients treated with ampulla local excision,162 patients treated with pancreaticoduodenectomy).Results For both ampullary cancer and benign tumor,the group of patients who received local excision was superior to the group of patients who received pancreaticoduodenectomy in operative time,blood loss,hospital stay,and incidence of postoperative complications (all P＜0.05).For benign ampullary adenomas,there were no significant differences in postoperative median survival time between local resection and pancreaticoduodenectomy (P=0.071,x2 =2.003).For stage T1 or T2 ampullary cancer,there was also no significant difference in the postoperative median survival time (P=0.054,x2=3.163).However,the postoperative median survival time of the pancreaticoduodenectomy group was significantly longer than that of the local resection group for patients with stage T3 or T4 ampullary cancer (P=0.041,x2 =6.309).Conclusions Ampullary tumors are characterized by relatively good response to treatment.The results suggested that for patients with benign ampullary adenomas and stage T1 or T2 of ampullary cancer,local resection is a good surgical procedure.For patients with ampullary cancer at more advanced stages (T3 or T4),pancreaticoduodenectomy is the first choice of therapy.

Objective To summarize the clinical and pathological characteristics of primary gallbladder carcinoma, analyze factors such as surgical procedures, tumor stages that associated with prognosis, and standardize the diagnosis and treatment strategy for primary gallbladder carcinoma.Methods From January 1986 to October 2009, 197 patients of primary gallbladder carcinoma were treated in Peking Union Medical College Hospital (PUMCH). A retrospective study was performed basing on clinical data of these patients, statistical analysis carried out using SPSS version 13.0, Kaplan-Meier estimate employed for the survival analysis and Log-rank test for the group comparisons. Results One hundred and ninety-seven patients were enrolled (73 male and 124 female) with the mean age of 64.4±1.1years. Abdominal pain (77.3%) , obstructive jaundice (33.6%) and fever (19.5%) were the main symptoms of primary gallbladder carcinoma, with accompanying cholecystolithiasis in 99 cases. B-mode ultrasonic examination and CT scan were important for preoperative diagnosis. Nevin stages and surgical modalities were independent prognosis factors. For stage Ⅰ and Ⅱ patients the survival terms were not different between those receiving radical cholecystectomy and simple cholecystectomy. However, for stage Ⅲ, Ⅳ and Ⅴ patients the survival terms were of statistically different between those in radical cholecystectomy group and simple cholecystectomy group. Conclusions For stage Ⅰ and Ⅱ patients, simple cholecystectomy is a good surgical procedure. For those at more advanced stages, radical cholecystectomy is the choice of therapy, while simple cholecystectomy also improves the prognosis compared with laparotomy only.