Objective The present study is to investigate the influence of vaginal discharge on dry chemistry determination of leukocyte esterase in female urine.Methods Collected 20 and 30 normal vaginal discharge samples.Which humoral routine test degree were Ⅰ and Ⅱ correspondingly,and then analyze leukocyte esterase and squamous epithelial cells in these normal vaginal discharge samples.Collected normal vaginal discharge samples again and isolate the vaginal squamous epithelial cells from those normal vaginal discharge samples.Made two concentration of squamous epithelial cell suspension liquid (20/μl and 60/μl)and added these liquid.To normal female urine and analyze the drying chemical leukocyte esterase in those urine. Cleaned the vulva of those patients with normal leukocyte counts in urine sediment determination,whose dry chemical deter-mination of leukocyte esterase was strong positive,perform the routine urine test with her middle urine.Results The results of the determination of leukocyte esterase in normal vaginal discharge samples were 66.7% positive.And there were ~squamous epithelial cells in all samples (microscopy).No significant difference was observed in the examination of dry chem-istry leukocyte esterase among the normal female urine group,low (20/μl)and high (60/μl)concentrations of squamous epi-thelial cell urine group (P >0.05).The counts of squamous epithelial cells and the rate of positive and intensity of dry chem-ical leukocyte esterase in the middle of second urine were significantly lower than before,and the differences are statistically significant (P <0.05).But there was no significant difference for leukocytes counts.Conclusion Urine squamous epithelial cells had no influence on the detection of leukocyte esterase by dry chemistry.However,the leukocyte esterase in the vaginal discharge greatly influenced the examination of the leukocyte esterase in urine.

ObjectiveTo investigate the learning curve of total thoracoscopicy cardiac surgery.MethodsClinical data of a succession of 125 patients undering total thoracoscopicy ASD and VSD repair between October 2004 to January 2010 were collected and reviewed.The procedure was perfomed by the same surgeon.The patients were divided equally into 5 groups (groupA,B,C,D and E,n =25 in each group ) according to the sequence of the operation.The operative time,extracorporeal circulation time,aortic cross-clamped time,the rate of conversion rate to thoracotomy and postoperative complications were compared between the 5 groups.ResultsThere were no statistically significant differences between the 5 groups with respect to age,gender,weight,dieases and surgical approach(P ＞ 0.05).The operative time,extracorporeal circulation time and aortic clump time in group A and group B significant longer then that in group C,group D and group E(P ＜0.05).Group A and group B was no statistically significant difference each other( P ＞ 0.05 ).Group C,D and E have no statistically significant differences between(P ＞0.05 ).The rate of conversion rate to thoracotomy and postoperative complications in 5 groups have no statistically significant differences comparative( P ＞ 0.05 ).ConclusionThe learning curve of total thoracoscopicy surgery is approximalely 50 cases.

BACKGROUND:Lung cancers are highly heterogeneous and resistant to available therapeutic agents, with a five year survival rate of less than 15%. It has been difficult to determine the basis of lung cancer heterogeneity and drug resistance. Cancer stem cellmodel has attracted a significant amount of attention in recent years as a viable explanation for the heterogeneity, drug resistance, dormancy and recurrence and metastasis of various tumors. OBJECTIVE:To summarize the current understanding of lung cancer stem cells, including their histological types and tumor growth areas, and to discusses the prognosis of lung cancer and its relationship with lung cancer stem cells, in an effort to eradicate these cells to combat lung cancer. METHODS:In order to search relevant articles about the lung cancer stem celland its relationship with lung cancer from PubMed and Sciencedirect databases (from 1990 to 2014), a computer-based search was performed, using the key words of“lung cancer, cancer stem cell, lung cancer stem cell, lung cancer occur, tumor heterogeneity, drug resistance, gene mutation, signal pathways”in English. After eliminating literatures which were irrelevant to research purpose or containing a similar content, 48 articles were chosen for further analysis. RESULTS AND CONCLUSION:The cancer stem cellmodel has gained considerable support recently in context of lung cancers and stem-like cells that are associated with aggressive cancer behavior, metastatic progression, resistance to therapy and relapse. Since lung cancer stem cells are thought to consist of a heterogeneous population depending on the histology and site of tumors, and multiple signaling pathways might have to be targeted to effectively eliminate lung cancer stem cells for therapeutic benefit. It can be imagined that the multidisciplinary efforts currently under way to characterize and target stem-like cells in lung cancer wil reap significant therapeutic benefits in the future.

Objective The individualized treatment of breast cancer have attracted more and more attention .Different mo-lecular subtypes of breast cancer have different kinds of prognosis and therapeutic regimen .Studies have found that miR-342-3p is asso-ciated with breast cancer of hormone receptor and endocrine therapy resistance , as well as tumor cell apoptosis .This study was to fur-ther investigate the expressions of miR-342-3p in different breast cancer molecular subtypes and breast cancer cell lines to reveal the importance of miR-342-3p in individualized treatment of breast cancer . Methods A total of 90 tissue samples from patients with breast cancer surgery were collected .Three types of breast cancer cell line were cultured , including MCF-7, SKBr3 and MDA-MB-231.Real-time fluorescent quantitative PCR was applied to detect the expression of miR-342-3p in breast cancer tissue . Results Expression of the miR-342-3p increased the most in Lumina B type breast cancer tissue (1.594 ±0.465), followed by Lumina A type (1.386 ±0.443), Her-2 high expression type (1.165 ±0.337), and the lowest in the tripe negative breast cancer tissue (0.837 ± 0.351), representing significant difference (P<0.05).There was no statistical difference in the expression of the miR-342-3p as to different age groups, lymph node metastasis and tumor size, histological grading and staging (P>0.05).As to the expression of miR-342-3p in three types of breast cancer cell line , taking SKBr3 as the reference, the relative ratio was 126(118-134) and MDA-MB-231 was 0.017(0.014-0.018). Conclusion The expressions of miR-342-3p are different in different molecular subtypes and cell lines of breast cancer , which are relevant to different molecular subtypes of breast cancer , making it possible reference index for breast cancer typing and relevant to good prognosis .

Objective To illustrate the clinical relevance of distinct PML-RARα fusion gene isoforms in acute promyelocytic leukemia (APL). Methods The nested reverse transcriptase polymerase chain reaction (RT-PCR) was used to detect the long (L) or short (S) PML-RARα fusion gene isoforms in 92 newly diagnosed APL so as to evaluate the clinical feature, therapeutic reaction and prognosis of the two fusion gene isoforms. Results PML-RARα fusion gene was positive in all 92 APL patients, of which 52(56.5 %) was L type and 40 (43.5 %) was S type. There were no significant differences between L type and S type in the aspect of sex, age, white blood cell count,the percentage of bone marrow blasts plus promyeloeytes and chromosome before treatment. And there were no significant differences between the two isoforms in complete remission (CR) rate, the time of getting CR as well as the occurrence of retinoic acid syndrome (RAS), disseminated intravascular coagulation (DIC), intraeranial hemorrhage. Also, there were no significant differences in overall survival rate (OS) and relapse-free survival rate (RFS) between the two isoforms. Conclusion PML-RARα fusion gene isoforms in APL were not correlated with clinical therapeutic effect or prognosis.

BACKGROUND:Studies have shown that lung cancer stem cels can be isolated from the lung cancer cel lines, But there are few reports on in vitro isolation, culture and identification of lung cancer stem cels in patients with lung squamous carcinoma.
OBJECTIVE:To establish the feasible methods of harvesting lung cancer stem cels from fresh lung cancer tissues in patients with lung squamous carcinoma, and to investigate the alterations in cel number and function during primary culture.
METHODS: Side population cels were isolated by colagenase digestion, Ficol density gradient centrifugation and Hoechst 33342 efflux properties. The isolated cels were isolated and cultured in conditioned medium. Flow cytometry method was used to detect lung cancer stem cels based on the cel surface markers CD133 and CD44, and the positive rates of CD133+, CD44+ and CD133+/CD44+ were recorded. The single cel clones assay, flat colony formation assay and the cel sphere formation assay were used to identify the stem-like characteristics of lung cancer stem cels between the first and fourth generations.
RESULTS AND CONCLUSION:The positive rates of CD133+, CD44+ and CD133+/CD44+ cels at the fourth generation were increased significantly, and the positive rates of CD133+ and CD133+/CD44+ cels at passage 4 were significantly higher than those at the first generation. The abilities of single cel clone formation, the flat colony formation and the cel sphere formation in the fourth-generation cels were greatly enhanced compared with the first-generation cels. Experimental findings showed that stem cel-like lung cancer cels were obtained from fresh lung cancer tissue in patients with lung squamous carcinoma, which stably and rapidly amplified in vitro, laying the foundation for the further study of the heterogeneity and drug resistance of lung cancer stem cels.

BACKGROUND:Studies have shown that lung cancer stem cel s can be isolated from lung cancer cel lines. But there are few reports about in vitro isolation, culture and identification of lung cancer stem cel s in patients with lung squamous carcinoma.
OBJECTIVE:To explore the feasible methods of harvesting lung cancer stem cel s from fresh lung cancer tissue in patients with lung squamous carcinoma.
METHODS:Side population cel s were isolated by col agenase digestion, Ficol density gradient centrifugation and Hoechst 33342 solution. The isolated cel s were suspended in conditioned medium for isolated culture. Flow cytometry method was used to detect lung cancer stem cel s based on the cel surface markers CD133 and CD44, and the positive rates of CD133+, CD44+and CD133+/CD44+cel s were recorded.
RESULTS AND CONCLUSION:Cel s adhered at 0.5 hour after incubation;typical cel colony was formed at 4 days of culture;cel s showed paving stone-shape at 7 days in a total number of 10 8. The positive rates of CD133+, CD44+and CD133+/CD44+cel s at passage 4 were increased significantly. These findings indicate that stem cel-like lung cancer cel s were obtained from fresh lung cancer tissue in patients with lung squamous carcinoma, which were stably and rapidly amplified in vitro, laying the foundation for the further study on the heterogeneity and resistance of lung cancer stem cel s in the future.

Objective To construct a recombinant plasmid encoding Tp0821,a membrane lipoprotein of T. pallidum,express and purify this protein,and to evaluate its immunocompetence.Methods The recombinant plasmid pQE32/Tp0821 was constructed and induced to express the corresponding protein.Then,New Zealand rabbits were immunized with purified recombinant protein to prepare polycional antibodies,and the titer of polyclonal antibody was determinated.Indirect ELISA was developed with the recombinant protein of T. pallidum as coating antigen to detect 80 control sera and 150 FTA-ABS-positive sera.Results The recombinant plasmid pQE32/Tp0821 was constructed and a fusion protein with expected molecular weight was expressed.Specific humoral response was elicited by the recombinant protein in New Zealand rabbits and the antibody titer reached 1:6400.Compared with FTA-ABS test,the indirect ELISA showed a sensitivity and specificity of 92.6%and 98.6%,respectively,in the detection of control and clinical sera.Conclusion The recombinant protein Tp0821 shows excellent immunocompetence,which can be applied to the serological diagnosis of syphilis.

Objective To investigate the factors associated with postoperative metastasis, recurrence, and survival in patients with stage ⅢA ( N2 ) non?small cell lung cancer ( NSCLC ) , and to provide an objective basis for postoperative radiotherapy in patients. Methods Clinical data were collected from 199 patients who underwent complete resection and were diagnosed with stage ⅢA ( N2 ) NSCLC after surgery in our hospital from 2009 to 2013. The Cox regression model was used for the multivariate analyses of metastasis and recurrence. The survival rates were calculated using the Kaplan?Meier method and analyzed using the log?rank test. Results In the 199 patients, 173 had complete follow?up data. The 1?and 2?year metastasis, recurrence, and survival rates were 38?7%/52?6%, 27?8%/39?1%, and 92?5%/51?4%, respectively. The multivariate analysis showed that pathological type and two positive indices among preoperative CEA/CY211/SCC were two risk factors for metastasis ( P=0?013,0?014) . Positive lymph node number, metastatic lymph node number, lymph node metastasis rate, and two positive indices among preoperative CEA/CY211/SCC were risk factors for recurrence ( P=0?046,0?004,0?028,0?001) . All the above indices were risk factors for low survival rates ( P= 0?013 , 0?011 , 0?002,0?026 ) . Conclusions Patients with stage ⅢA ( N2 ) NSCLC who have positive lymph nodes, lymph node metastases, and two positive indices among preoperative CEA/CY211/SCC may benefit from postoperative radiotherapy.

Objective To clone, express Tp0319 gene from Treponemapallidum (T. pallidum), and to assess the immunocompetence of recombinant protein. Methods The immuno-dominant region of Tp0319gene was chosen by computer analysis, amplified from T. pallidum complete genome by PCR, subcloned into the expression vector pQE32 to construct a recombinant plasmid, pQE32/Tp0319, which was then expressed in E. coli M15. The recombinant protein was purified with Ni-NTA affinity chromatography, and identified by using sodium dodecyl sulfate polyacrylamide gel electropheresis (SDS-PAGE) and Western blot. New Zealand rabbits were immunized with the recombinant protein, and the titer of anti-Tp0319 antibodies in sera from immunized rabbits were measured with indirect ELISA. Also, indirect ELISA with the recombinant Tp0319 as coating antigen was performed to detect the anti-Tp0319 antibody in sera from 200 normal human controls and 200 patients with syphilis. Results The prokaryotic expression vector pQE32/Tp0319 was constructed successfully, and the recombinant protein Tp0319 with a molecular weight of about 30 000 was attained. Specific humoral response was elicited by the recombinant protein in New Zealand rabbits and the specific antibody titer was more than 1: 10 240 after immunization for 3 times. Western blot proved that the recombinant protein could specifically react with anti-T. pallidum IgG antibody-positive sera. Indirect ELISA was successfully developed with the recombinant Tp0319, and detected antibodies to T. pallidum in control sera with a sensitivity and specificity of 100% (40/40), respectively. Compared with T. pallidum particle agglutination (TPPA) assay, the sensitivity and specificity of the indirect ELISA were 92.6% and 100%, respectively, in the detection of T. pallidum in sera from patients and controls, and the concordance between the indirect ELISA and TPPA was 96%. Conclusions The prepared recombinant protein shows a satisfactory immunocompetence, which may lay a foundation for its further application in the serodiagnosis of syphilis.