Objective:To explore relation of TCM syndrome types in patient with sexual hormones of hyperplasia of mammary glands,so as to establish objective indexes of TCM syndrome types of hyperplasia of mammary glands.Methods:112 cases of hyperplasia of mammary glands were divided into 3 types:stagnation of liver-Qi,intermingled phlegm and blood stasis,disharmany of thoroughfare vessel and conception vessel.Level of estradiol(E_2),progestin(PT),testosterone(TT),prolactin(PRL),follicle- stimulating hormone(FSH)and luteotropic hormone(LH)during ovulatory period in the patients were determined.The relation of hormone level with TCM syndrome types was analyzed.Results:Level of LH and E_2 decreased,PRL and PT increased and level of TT and FSH did not significantly change during ovulatory period in the patient of hyperplasia of mammary glands.LH and E_2 level in the patients with stagnation of liver-Qi or intermingled phlegm and blood stasis was significantly lower than normal values,and the level of PRL and PT was significantly higher than that of normal values.The level of LH and E_2 was also lower than the normal value in the patient with disharmany of thoroughfare vessel and conception vessel,but the level of FSH was significantly higher than normal value.Conclusion:The patient of hyperplasia of mammary glands during ovulatory period manifests endocrine dysfunction,and the patients with different TCM syndrome are different in secretion of sexual hormones; Changes of sexual hormones can be used as objective indexes of TCM syndrome types and basis of clinical medicinal application.

Objective To analyze the clincopathological features and prognosis of 77 colorectal cancer patients with synchronous liver metastases after resection of the primary tumors.Methods Seventy seven colorec-tal cancer patients with synchronous liver metastases who underwent resection of the primary tumors and patholog-ical confirmed were enrolled in this retrospective study.The clincopathological features and prognosis were ana-lyzed.Kaplan-Meier method was used to analyze survival rate.The differences between survival curves were cal-culated by Log-rank test.Results The analysis showed that sex,tumor differentiation and TNM stage were as-sociated with prognosis(P<0.05).The analyses demonstrated no differences in age,primary tumor site,treat-ment,number of liver metastases and extrahepatic metastasis(P>0.05).There was a significant correlation be-tween CEA,CA19-9 level and gender,tumor differentiation and TNM stage(P<0.001)and the corresponding surgical procedure and TNM stage(P<0.05).Conclusion The analysis show that male,the low differentiation and stageⅢare poor prognosis factors.

ObjectiveTo investigate the effect of the expression of HBcAg in hepatocytes on the serum level of HBcAb and seroconversion of HBeAg after antiviral therapy with nucleos(t)ide analogues (NUCs). MethodsSerum samples and liver tissue paraffin sections were collected from 101 chronic hepatitis B (CHB) patients who received antiviral therapy with NUCs in Nanfang Hospital and Panyu Central Hospital from January 2015 to June 2018. ELISA was used to measure the serum level of HBcAb, and immunohistochemistry was used to measure the expression of HBcAg in the liver. The GEO database (GSE96851) was analyzed to obtain differentially expressed genes in the liver of patients with HBcAg-positive hepatitis. The two-independent-samples t test was used for comparison of continuous data between two groups; the multiple-independent-samples nonparametric Kruskal-Wallis H test was used for comparison of continuous data between multiple groups, and Dunnett method was used for further comparisons; the chi-square test was used for comparison of categorical data between groups. ResultsThe expression pattern of HBcAg in hepatocytes was classified as absent expression, nuclear expression, cytoplasmic expression, and nuclear/cytoplasmic expression, and according to expression level, HBcAg expression was classified as grades Ⅰ, Ⅱ, Ⅲ, and Ⅳ expression. HBeAg seroconversion rates after 96 weeks of antiviral therapy were 5.88%, 16.67%, 22.73%, and 24.24%, respectively, in the patients with absent expression, nuclear expression, cytoplasmic expression, and nuclear/cytoplasmic expression (χ2=4753, P=0.037), and HBeAg seroconversion rates after 96 weeks of antiviral therapy were 5.88%, 13.04%, 27.59%, and 26.67%, respectively, in the patients with grade Ⅰ, Ⅱ, Ⅲ, and Ⅳ expression (χ2=6.580, P=0.016). There were significant differences in the serum levels of HBcAb-IgM and total HBcAb between the patients with absent expression, nuclear expression, cytoplasmic expression, and nuclear/cytoplasmic expression of HBcAg (HBcAb-IgM: H=9.760, P=0.021; total HBcAb: H=21.46, P＜0.001), and there were also significant differences in the serum levels of HBcAb-IgM and total HBcAb between the patients with grade Ⅰ, Ⅱ, Ⅲ, and IV expression of HBcAg (HBcAb-IgM: H=18.80, P＜0.001; total HBcAb: H=26.03, P＜0.001). The analysis of differentially expressed genes in the liver showed that the expression of antibody-related genes was upregulated in the liver of patients with HBcAg-positive acute liver failure. ConclusionThe expression pattern and level of HBcAg in the cytoplasm of hepatocytes are associated with serum HBcAb, and the measurement of HBcAg may help to predict the efficacy of antiviral therapy with NUCs.

OBJECTIVETo construct full-length human bladder cancer-specific antibody libraries for efficient display of full-length antibodies on the surface of mammalian cells.

METHODSThe total RNA was isolated from peripheral blood mononuclear cells from patients with bladder cancer. The repertoires of IgG1 heavy chain variable region (VH) and Kappa light chain were amplified by RT-PCR using specific primers. The antibody genes were inserted into the vector pDGB-HC-TM to construct the bladder-cancer-specific antibody libraries of heavy chains and light chains. Ten clones from each library were randomly picked for gene sequencing and transient transfection into FCHO cells to analyze antibody display on mammalian cell surface by flow cytometry after staining with corresponding fluorescent labeled antibodies.

RESULTSThe libraries of bladder-cancer-specific antibody heavy chain (IgG1) and light chain (LCk) were successfully constructed. Seven out of the 10 clones randomly selected from the heavy chain library and 9 out of the 10 clones from the light chain library showed correct open reading frame, coding for 7 unique VH and 9 unique LCk. The combinatory library size reached 3.32×10(11).

CONCLUSIONWe have successfully constructed a full-length human bladder-cancer-specific antibody library with a combinatory diversity of 3.32×10(11) based on mammalian display technology, which can be used for screening monoclonal antibodies against bladder-cancer-associated antigens.

Amino Acid Sequence ; Animals ; Antibodies ; genetics ; Cell Surface Display Techniques ; Gene Library ; Humans ; Immunoglobulin Heavy Chains ; genetics ; Immunoglobulin kappa-Chains ; genetics ; Peptide Library ; Urinary Bladder Neoplasms ; genetics ; immunology

Objective:To investigate clinical efficacy of conformal sphincter preservation operation (CSPO) versus intersphincteric resection (ISR) in the treatment of low rectal cancer.Methods:The retrospective cohort study was conducted. The clinicopathological data of 183 patients with low rectal cancer who were admitted to two medical centers (117 in the Changhai Hospital of Naval Medical University and 66 in the Huashan Hospital of Fudan University) from August 2011 to April 2020 were collected. There were 110 males and 73 females, aged (57±11)years. Of 183 patients, 117 cases undergoing CSPO were allocated into CSPO group, and 66 cases undergoing ISR were allocated into ISR group, respectively. Observation indicators: (1) surgical situations of patients with low rectal cancer in the two groups; (2) postoperative complications of patients with low rectal cancer in the two groups; (3) follow-up; (4) influencing factors for prognosis of patients with low rectal cancer; (5) influencing factors for satisfaction with the anal function of patients with low rectal cancer. Follow-up was conducted using outpatient examination, questionnaire and telephone interview to determine local recurrence, distal metastasis, survival, stomal closure, satisfaction with the anal function of patients. Measurement data with normal distribution were represented as Mean±SD, and comparison between groups was analyzed using the t test. Measurement data with skewed distribution were represented as M (range). Count data were described as absolute numbers or percentages, and comparison between groups was analyzed using the chi-square test. Comparison of ordinal data was analyzed using the rank sum test.The Kaplan-Meier method was used to draw survival curves, and life table method was used to calculate survival rates. Log-rank test was used for survival analysis. Univariate analysis was performed using the linear regression. Variables with P<0.10 in the univariate linear regression analysis were included for multivariate analysis. Multivariate analysis was performed using the COX stepwise regression model and linear regression analysis. Results:(1) Surgical situations of patients with low rectal cancer in the two groups: cases with laparoscopic surgery, operation time, volume of intraoperative blood loss, distance from tumor to distal margin, cases with postoperative chemotherapy, duration of postoperative hospital stay were 44, (165±54)minutes, (142±101)mL, (0.6±0.4)cm, 76, (6.6±2.5)days for the CSPO group, respectively, versus 55, (268±101)minutes, (91±85)mL, (1.9±0.6)cm, 9, (7.9±4.7)days for the ISR group, showing significant differences between the two groups ( χ2=35.531, t=8.995, -3.437, -3.088, χ2=44.681, t=2.267, P<0.05). (2) Postoperative complications of patients with low rectal cancer in the two groups: 19 patients in the CSPO group had complications. There were 6 cases with grade Ⅰ complications, 12 cases with grade Ⅱ complications, 1 case with grade Ⅲb complication. Fourteen patients in the ISR group had complications. There were 4 cases with grade Ⅰ complications, 7 cases with grade Ⅱ complications, 1 case with grade Ⅲa complication, 2 cases with grade Ⅲb complications. There was no significant difference in the postoperative complications between the two groups ( χ2=0.706, P>0.05). Patients with complications in the two groups were improved after symptomatic and supportive treatment. There was no perioperative death in the postoperative 30 days of the two groups. (3) Follow-up: 183 patients received follow-up. Patients of the CSPO group and ISR group were followed up for (41±27)months and (37±19)months, respectively, showing no significant difference between the two groups ( t=-1.104, P>0.05). There were 2 cases with local recurrence and 9 cases with distal metastasis of the CSPO group, respectively, versus 3 cases and 4 cases of the ISR group, showing no significant difference between the two groups ( χ2=1.277, 0.170, P>0.05). The 3-year disease-free survival rate and 3-year total survival rate were 84.0% and 99.0% for the CSPO group, versus 88.6% and 92.8% for the ISR group, showing no significant difference between the two groups ( χ2=0.218, 0.002, P>0.05). The stomal closure rate was 92.16%(94/102) and 96.97%(64/66) for 102 patients of CSPO group and 66 patients of ISR group up to postoperative 12 months,respectively, showing no significant difference between the two groups ( χ2=1.658, P>0.05). Of the 8 cases without stomal closure in the CSPO group, 2 cases refused due to advanced age, 4 cases subjectively refused, and 2 cases were irreducible due to scar caused by radiotherapy. Two cases in the ISR group had no stomal closure including 1 case of postoperative liver metastasis and 1 case of subjective refusal. There were 92 and 61 patients followed up to 12 months after stomal closure, of which 75 cases and 38 cases completed questionnaires of satisfaction with the anal function. The satisfaction score with the anal function was 6.8±2.8 and 5.4±3.0 for CSPO group and ISR group, respectively, showing a significant difference between the two groups ( t=-2.542, P<0.05). Fifty-four cases in the CSPO group and 21 cases in the ISR group had satisfaction score with the anal function >5, showing no significant difference between the two groups ( χ2=3.165, P>0.05). (4) Influencing factors for prognosis of patients with low rectal cancer: results of COX stepwise regression analysis showed that gender and pT staging were independent influencing factors for disease-free survival rate of patients with low rectal cancer ( hazard ratio=2.883, 1.963, 95% confidence interval as 1.090 to 7.622, 1.129 to 3.413, P<0.05). Gender and pT staging were independent influencing factors for total survival rate of patients with low rectal cancer ( hazard ratio=10.963,3.187, 95% confidence interval as 1.292 to 93.063, 1.240 to 8.188, P<0.05). (5) Influencing factors for satisfaction with the anal function of patients with low rectal cancer: results of univariate analysis showed that surgical method and tumor differentiation degree were related factors for satisfaction with the anal function of patients with low rectal cancer (partial regression coefficient=1.464, -1.580, 95% confidence interval as 0.323 to 2.605, -2.950 to -0.209, P<0.05). Results of multivariate analysis showed that surgical method, tumor differentiation degree and preoperative radiotherapy were independent influencing factors for satisfaction with the anal function of patients with low rectal cancer (partial regression coefficient=1.637, -1.456, -1.668, 95% confidence interval as 0.485 to 2.788, -2.796 to -0.116, -2.888 to -0.447, P<0.05). Conclusion:Compared with ISR, CSPO can safely preserve the anus in the treatment of low rectal cancer, without increasing the incidence of postoperative complications, which can also guarantee the oncological safety and improve the postoperative anal function.