Objective To study the interaction between HCV core protein and HCBP6 in mammlian cells using CheckMateTM Mammalian Two-Hybrid System.Methods cDNA fragments encoding HCV core protein and HCBP6 were amplified by PCR and subsequently cloned into pGEM-T vector.After verified by sequencing,the target fragments were subcloned into mammalian two-hybrid plasmids,pBIND and PACT,respectively.The recombinant plasmids,pBIND-core and pACT-HCBP6 were co-transfected into HepG2 cells with reporter plasmid pG5luc.pBIND+pACT were induced as background controls,pBIND-Myod+pACT-Id as positive controls,and pBIND-core+pACT,pBIND+pACT-HCBP6 as blank controls.The expression of G5luc,which indicated the interaction between HCV core protein and HCBP6 in mammlian HepG2 cells,was assayed through Dual-Luciferase Report Assay System and Turner Biosystems Veritas Microplate Lunimometer.Results The recombinant vectors pBIND-core and pACT-HCBP6 were successfully constructed.When co-transfected into HepG2 cells with reporter plasmid pG5luc,there were significant differences in the luciferase activity in the pBIND-core and pACT-HCBP6 groups compared with every control group.Conclusions HCBP6 can interact with HCV core protein in HepG2 cells,which provides clues for further study on the function of HCBP6 and core proteins,and on the mechanism of HCV core in cell apoptosis and cancer transformation.

Objective: To evaluate reliability and validity of ATQ. Methods: A total of 350 undergraduates and 102 Psychiatric patients were tested by ATQ and BDI. Results:The ATQ attained good psychometric properties: Cronbach α of ATQ was 0.95, spit-half correlations ranged from 0.90 to 0.94; correlation coefficient between ATQ and BDI was 0.54 for normal undergraduates, 0.60 for schizophrenics, and 0.75 for depressive subjects (p<0.001). There were significant difference among non-depressed undergraduates、depressed undergraduates、schizophrenics and depressive patients. Conclusion: The present study provided empirical support for the reliability and validity of ATQ.

Objective To investigate the clinical features of gynecological malignant tumor related multiple primary malignant neoplasms (MPMN).Methods Apply retrospective and comprehensive analysis to the clinical data of 30 patients with gynecological malignant tumor related MPMN.Results Synchronous MPMN were found in 9 patients.Their average age was 50.2 years old and their median age was 49 years old.The neoplasms were located at ovary,uterus,cervix,breast and intestine.Metachronous MPMN were found in 21 patients.Their average age was 57.7 and their median age was 57 years old.The median interval between the first and the second primary malignant neoplasm was 4.0 years.The neoplasms were located at breast,ovary,uterus,gastrointestinal tract,uterine cervix,lung etc.In 30 cases,26 of them were treated by surgical operation and further adjunctive treatment of chemotherapy and (or) radiotherapy was conducted as per the neoplasm staging and its pathological results.The rest 4 patients (first primary malignant neoplasms were excised from 3 of them and another one was not treated by surgical operation) received adjunctive treatment of chemotherapy and (or) radiotherapy.Followed ups,which varied from 6 to 60 months,were made to 29 patients and 20 out of the 29 were alive.5-year survival rate of patients with gynecological malignant tumor related MPMN was 47.8％,2-year survival rate was 73.9％,and 1-year survival rate was 88.6％.Conclusion Pay more attention to the patients with gynecological malignant tumor related MPMN,examine the high-risk patients with malignant tumor comprehensively,identify whether it is recurrence,metastasis or new growth of malignant neoplasm,and further ensure early diagnosis and proper treatment,avoiding misdiagnosis and missed diagnosis.

Objective To prepare polyclonal antibody of transactivated protein 5 of hepatitis C virus nonstructural 5A (NA5ATP5) and to explore its expression in the liver tissues. Methods In Escherichia coil BL21, the prokaryotic expression vector pET32a(+)-NS5ATP5 was induced by isopropyl-β-D-thiogalactoside (IPTG), and it was analyzed with sodium dodecyl sulfate-polyaerylamide gel electrophoresis (SDS-PAGE) and Western blotting. And the purified protein was used to immunize the rabbit to prepare polyelonai antibody, with which we studied the function of NSSATP5 by determining the different liver tissues with the streptavidin-perosidase (SP) immunohistochemistry method. Results Recombinant NS5ATP5 (molecular weight: 65 kD) and polyclonal antibody were successfully prepared. NS5ATP5 expression in the liver of patients with chronic HCV infection was much higher than that of a normal person, and it was detected mainly in the cytoplasm. Conclusion The findings of the expression difference between HCV patients and normal people led to a novel diagnostic marker to detect HCV infection.

OBJECTIVE: To investigate the sleep monitoring feature of the MSMS in elderly patients with OSAHS. METHOD: One hundred and ninety patients diagnosed with OSAHS were divided into elderly group and non elderly group according to age, then the results of MSMS were analyzed. RESULT: Majority elderly patients were with mild to moderate OSAHS. The nocturnal mean blood oxygen and the lowest oxygen were higher than non elderly group, coupled with higher percentage of the total oxygen saturation < 90% monitoring time (TS90). There was no significant difference in sleep structure between two groups, but the total sleep time of elderly group is lower than the non elderly group, the difference is statistically significant. CONCLUSION The elderly patients with OSAHS were less severe in nature, but the nocturnal hypoxia last longer in the elderly group. There is no significant difference in the sleep structure between the two groups. But the total sleep time decrease in elderly group. With smaller interference, the MSMS is closer to the natural sleep stustus of the subjects.
Aged ; Humans ; Hypoxia ; diagnosis ; Oxygen ; physiology ; Polysomnography ; Sleep ; Sleep Apnea, Obstructive ; diagnosis

Objective To investigate the efficacy and side effects of the molecular targeted drug sunitinib in the treatment of metastatic renal cell carcinoma(mRCC). Methods Fifteen patients with histopathological confirmed mRCC,including 10 males and 5 females,were enrolled in the study.The median age was 56(range from 37 to 73 years).Fourteen cases of clear-cell RCC and 1 papillary RCC were diagnosed.Thirteen patients had prior nephrectomy,and 6 patients were treated with cytokines previously.All of the patients were given sunitinib at a dosage of either 50 mg daily(4 weeks on/2 weeks off)or 37.5 mg daily continuously.Objective response rate(ORR),progressive-free survival(PFS)and overall survival(OS)were evaluated,and adverse events were also observed. Results The median follow-up was 13 months(2-24 months).According to RECIST,partial response could be evaluated in 8 patients and stable was found in another 5 patients.Only 2 patients were confirmed disease progression.The ORR was 53% with the disease control rate of 87%,However,the median PFS and OS were not yet available due to the short-term follow-up.During the treatment,the most common adverse events experienced by patients were hand-foot syndrome 11/15(73%),changes in hair color 10/15(67%),mucositis 9/15(60%),alopecie 9/15(60%),diarrhea 8/15(53%)and neutropenia 8/15 (53%). All of adverse events were manageable and reversible. Conclusions Sunitinib as a single agent in the treatment for Chinese mRCC patient is efficient and the side effects are reversible. Further long-term follow-up and expanded samples should be expected to confirm the efficacy and safety of sunitinib.

Objective To investigate radiation risk perception and nuclear acceptance after the 60Co stuck source incident in June 2009 in Qixian County,Henan Province.Methods From March to April 2010,23 hamlets and 2 schools from 5 towns in Qixian County were selected,while 3 hamlets and 2 schools from 3 towns were used as control in Huixian County,1 35 km away where no any impact of the incident was found.Face-to-face questionnaire survey was conducted among three groups including mass public,elite public(representatives of public opinion,such as physicians,teachers,govemmental officials,administrative personnel,private entrepreneurs),and middie school students(the third grade students in junior middle schools and the first grade students in senior middle schools).Results A total of 1 340 valid questionnaires were collected.The public knowledge levels about radiation and nuclear power were low among all groups in two areas and public knowledge levels in Qixian County were higher than those in Huixian County.In both areas.the knowledge levels of the students were higher tIIan those among the elite public,and that among the general public were the lowest.The knowledge levels of males and those with higher educational levels were higher than those of females and those with lower educational levels.More than 40.0% of the respondents supported the idea to build nuclear power plants in China.but only a small part of them supported to build nuclear power plants in their own areas.The percentages of opponents against local nuclear power plant construction were especially higher among the students and those with higher educational levels.More than 60% of the mass public and elite public believed the explanation of the government.More than 80% of the respondents held negative attitudes towards the performance of the Qixian County Government in dealing with this incident.The student gave the lowest evaluation of the credibility and the specific performance of the local government,whereas the elite public placed themselves between the students and the mass public.The reasons why the local people left their homes to evade risk included rumors(67.2%),lack of radiation knowledge(61.9%),group psychology(59.2%),and failure of the government to inform the truth to the public in time(42.3%).Conclusions The public still fails to know enough about radiation and nuclear power.Although they trust the government,they are dissatisfied with the performance of the local government in dealing with the incident.The negative evaluation of the students is worth concerning.

Objective: Tumor markers have been widely used clinically. Detection of serum CA125 is one of the commonly used clinical methods for early screening and early diagnosis of epithelial ovarian cancer, but it is difficult to diagnose epithelial ovarian cancer with a single specific tumor marker. In this study, the combinatorial tumor marker detection method was used to compare the value of each tumor marker alone and different combinations in the diagnosis of epithelial ovarian cancer. Methods: The clinical data of patients with epithelial ovarian cancer (n=65) and ovarian benign disease (n=29) were collected. Multiple tumor marker protein chip was used to detect cancer antigen 125 (CA125), carbohydrate antigen 242 (CA242), alpha-fetoprotein (AFP), beta-human chorionic gonadotropin (β-HCG), carcinoembryonic antigen (CEA), cancer antigen 199 (CA199), neuron-specific enolase (NSE), Ferritin, cancer antigen 153 (CA153), and human growth hormone (HGH) serum levels, and to compare the differences between the benign and malignant ovarian tumors. The correlation between tumor markers and clinicopathologic features for ovarian epithelial carcinoma was analyzed by χ2 test. Spearman rank analysis showed the correlation between CA125 expression level and other tumor markers in epithelial ovarian cancer and the correlation between age and the above 10 tumor markers. Sensitivity, specificity, positive predictive value, negative predictive value, Youden index, and diagnostic efficiency were used to evaluate the diagnostic value of single tumor marker and the combination of tumor markers. Results: The levels of β-HCG, NSE, CA153, and CA125 in the epithelial ovarian cancer group were higher than those in the ovarian benign disease group. The level of NSE in the serum of patients with epithelial ovarian cancer was related to the clinical stage of patients. In addition, the levels of CA242, β -HCG, CEA, NSE, Ferritin, CA153 in the serum of patients with epithelial ovarian cancer were positively correlated with CA125 (rs=0.497, P< 0.001; rs=0.612, P<0.001; rs=0.358, P=0.003; rs=0.680, P<0.001; rs=0.322, P=0.009; rs= 0.609, P<0.001, respectively), and the levels of β-HCG, Ferritin, CA153 were positively correlated with the patient's age (rs=0.256, P=0.040; rs=0.325, P=0.008; rs=0.249, P=0.046, respectively). In the diagnosis of epithelial ovarian cancer, the sensitivity, Youden index, and diagnostic efficiency of CA125 detection alone were higher than the results of the other 9 separate detections. When CA153, CA199, CA242, Ferritin, and CEA were combined with CA125, the sensitivity of the combined detection of different combinations was higher than that of CA125 alone. The combined detection sensitivities of CA125+CEA and CA125+Ferritin+CEA were 89.2％ and 90.8％, respectively, and the diagnostic efficiencies were both 84.1％, which were higher than those of other combinations. The Youden index of CA125+CEA joint detection was 0.616, which was higher than those of other combinations. Conclusion: CA125 has a high diagnostic value in the diagnosis of epithelial ovarian cancer. The detection of combined tumor markers in serum has higher sensitivity and specificity in epithelial ovarian cancer.

Objective:To explore the establishment and application of ovarian cancer organoids.Methods:Fresh ovarian tumor tissues, obtaining from patients underwent surgery in the First Affiliated Hospital of Nanjing Medical University between October 2021 and March 2022, were collected, enzymatic degraded, digested, and embedded into matrigel to establish organoids. A total of 32 ovarian cancer samples were collected. Hematoxylin eosin (HE) staining and immunofluorescence (IF) procedure were used to verify the morphological structure of organoids and their expression of molecular markers. 3D cyto-live or dead assay was used to detecte the live or dead cells in organoids. Carboplatin with a concentration ranging from 5 to 80 μmol/L (5, 10, 20, 40, 80 μmol/L) was added to organoids to calculate the 50% inhibitory concentration (IC 50) in different organoids. Results:(1) Organoids from a total of 32 patients were established, of which 18 cases could be passaged stably in the long term in vitro, while 14 could be passaged in the short time. The average amplification time of long-term passage in vitro was over 3 months, and the longest reached 9 months. (2) In HE staining, significant nuclei atypia and local micropapillary structures were observed in organoids. IF staining revealed that ovarian cancer organoids expressed molecular markers similar to primary tumor tissues, such as Pan cytokeratin (Pan-CK), p53, paired box gene 8 (PAX8), and Wilms tumor gene 1 (WT1). (3) In 3D cyto-live or dead assay, a large number of apoptotic cells were observed inside and around the organoids after added carboplatin. The sensitivity to carboplatin varied in 18 organoids could amplify in the long term, with an average IC 50 of (29.5±15.8) μmol/L. Moreover, IC 50 values of 4 organoids derived from patients received neoadjuvant chemotherapy were much higher than the 14 organoids which did not received neoadjuvant chemotherapy [(48.7±11.3) μmol/L vs (24.0±12.1) μmol/L; t=3.429, P=0.022]. Conclusions:Organoids recapitulate ovarian cancers in vitro and could be stably passaged. Organoids derived from patients received neoadjuvant chemotherapy have higher resistance to carboplatin.