Objective To obtain sufficient recombinant multiepitope peptide of ESAT-6.Methods DNA encoding ESAT-6 peptide was amplified by PCR using a pair of primers which were designed in accordance with the reported ESAT-6 gene sequence.The ESAT-6P gene was inserted into the prokaryotic expression vector pET-30a with the N-terminal 6His-tag.The recombinant peptide vector pET30a-ESAT-6P was transformed into E.coli BL21(DE3) via chemical transformation.The positive clone was screened by means of PCR.The target peptide was expressed in E.coli after induction with IPTG and analyzed by SDS-PAGE.The immunogenicity of the peptide was evaluated by dot immunobinding assay.Results The target peptide was successfully expressed and purified.The solubility analysis showed that the recombinant peptide existed as inclusion body in the E.coli.DIBA indicated that the ESAT-6P was specifically reactive to sera from TB patients.Conclusions The recombinant multiepitope peptide of ESAT-6 engineering bacteria has been obtained,which will be quite helpful to develop novel specific diagnostic reagents and effective vaccine.

Objective To discuss the clinical value of laparoscopic myomectomy.Methods We compared clinical data of laparoscopic myomectomy(Laparoscopic Group,n=185) with open myomectomy(Open Group,n=69),from December 2002 to February 2005,in respect of operative time,hemorrhage volume,time to normal temperature,recovery time of bowel movement,and duration of hospital stay.Results Conversions to open surgery were required in 2 cases in the Laparoscopic Group because of difficulties of hemostasis.There was no significant difference in the operative time(t=1.849,P=0.066) between the Laparoscopic Group(81.3?14.7 min) and the Open Group(77.2?18.1 min).The intraoperative hemorrhage volume was significantly less in the Laparoscopic Group(101.5?36.7 ml) than in the Open Group(154.5?61.1 ml)(t=-8.397,P=0.000).Shorter time to normal temperature,recovery time of bowel movement,and hospital stay were achieved in the Laparoscopic Group(2.7?0.8 d;13.8?5.4 d;6.0?1.5 d) than in the Open Group(3.8?1.0 d;23.3?6.0 d;8.0?2.0 d),with significant difference(t=-9.064,P=0.000;t=-12.074,P=0.000;t=-8.575,P=0.000).Follow-ups were conducted in 152 cases in the Laparoscopic Group for 13.2?8.7 months,which revealed 17 cases of recurrence(11.2%) and 17 cases of pregnancy within 1 year out of 57 cases of infertility(29.8%).As compared with the Laparoscopic Group,followups were conducted in 48 cases in the Open Group for(12.5?)7.9 months(t=0.511,P=0.610),which revealed 8 cases of recurrence(16.7%;?~2=1.003,P=0.317) and 4 cases of pregnancy within 1 year out of 16 cases of infertility(25.0%;?~2=0.004,P=0.949).Conclusions Laparoscopic myomectomy has advantages of micro-invasion,little blood loss,rapid postoperative recovery,and short hospitalization.However,it cannot entirely supersede the open myomectomy.

Objective To explore the continuous improvement to reduce the suctioning pediatrics lumen instruments return-cleaning rate of the first time washing, improve work efficiency and reduce the cost by applying root cause analysis. Methods Using causal analysis of fishbone diagram to analysis and verify the main reason of leading to high lumen instruments return-cleaning rate. According to the three terminal factors of continuous quality improvement, quality control group was set up, lumen instruments cleaning quality control standards was made, water flow mode of lumen instruments cleaning was changed, selected the appropriate cleaning tools and real picture show, synchronize quality control measures of publishing the quality and safety board. Compared before and after return-cleaning rate of three different detection methods and the different parts of the same suction lumen instruments. Results Before carrying out eye-measurement, cotton swab to wipe, ATP bioluminescence back washing rate was 0.89% (2/225), 7.11%(16/225), 27.11%(61/225), respectively after implementation of 0, 0.44%(1/226), 3.98%(9/226), visual observation before and after the return rate of washing was no statistically significant difference (χ2=2.018, P>0.05);Cotton swab to wipe, ATP bioluminescence back washing rate difference was statistically significant (χ2=13.820, 45.999, P<0.01). The lumen instruments total return-washing rate was decreased from 35.11% (79/225) to 4.42% (10/226). Among them, the return- washing rate of the inside surface of lumen instruments was decreased from 32.89% (74/225) to the 3.10% (7/226) and the difference was statistically significant(χ2=67.028, 67.915,P<0.01). By contraries, the thread interface and the outside surface of lumen instruments return- cleaning rate before and after the implementation has no statistical significance (P>0.05). Conclusions ATP bioluminescence assay has fine effects to detect the return-washing rate of the inner wall of the lumen instruments. The Root Cause Analysis method significantly reduced the return-washing rate of the inside surface of the suction lumen instruments, improve the efficiency, save the medical cost and reduce the hospital infection.

Objective To analysis the diagnostic values and characteristics of ultrasound and ultrasound guided fine needle aspiration cytology (FNAC) in patients with papillary thyroid carcinoma (PTC) before operation. Methods The data of ultrasound and ultrasound guided FNAC in 129 patients (including 148 PTC nodules) with PTC were collected from January 2014 to February 2017, and the diagnostic reports and feature descriptions of ultrasound and ultrasound guided FNAC were analyzed retrospectively. Results Among the 148 PTC nodules, the ultrasonographic imaging showed that 84.5% (125/148) with low echo-level solid nodules, 61.5% (91/148) with echo heterogenicity, 77.7% (115/148) with a ratio ≥ 1 in longitudinal/breadth, 69.6 % (103/148) with fuzzy boundary, 75.0% (111/148) with microcalcification in nodules, 97.3% (144/148) without or with incomplete aureole, 64.9% (96/148) with rich blood flow and 7.0% (9/129) with enlargement of cervical lymph nodes. The FNAC diagnosis showed that 78.4% (116/148) was diagnosed with suspected papillary carcinoma, 1.4% (2/148) was diagnosed with malignent tumor, 11.5% (17/148) was diagnosed with atypia of undetermined significance (AUS), 1.4% (2/148) was diagnosed with benign lesion, 0.7% (1/148) was diagnosed with follicular neoplasm and 6.7% (10/148) could not be diagnosed. If the suspected papillary carcinoma and malignent tumor were defined as cytodiagnosis, the diagnostic accordance rate with intraoperative pathology was 79.7%. Conclusion The preoperative accuracy rates of ultrasound diagnosis and ultrasound guided FNAC diagnosis for patients with PTC are high, and the characteristics of the both are also typical. The two examinations before operation are helpful for early diagnosis and treatment formulation for patients with PTC.

AIM: To investigate the effects of TNF receptor-associated death domains (TRADD) of latent membrane protein 1 (LMP1) on the proliferation of nasopharyngeal cancer SP18 cells.METHODS: The SP18-LMP1 cells and SP18-LMP1TRADD cells, which expressed LMP1 and LMP1 TRADD proteins, respectively, were established.The proliferation of SP18 cells affected by LMP1TRADD was detected by cell counting to analyze the cell growth curve, and by colony formation assay, soft agar formation assay, and flow cytometry.Moreover, the expression profile of differential genes between SP18-LMP1 cells and SP18-LMP1TRADD cells was analyzed by gene chips.RESULTS: The cell growth curve, and the results of colony formation and soft agar formation displayed that the growth velocity and colony forming ability of SP18-LMP1 cells were stronger than those of SP18-LMP1TRADD cells (P<0.01).The results of flow cytometry analysis showed that the proliferation index of SP18-LMP1 cells was higher than that of SP18-LMP1TRADD cells (P<0.01).Sixty-three differentially expressed genes associated with cell proliferation were screened out, in which 33 genes were up-regulated and 30 were down-regulated in the SP18-LMP1TRADD cells.CONCLUSION: TRADD active region is an important functional site of LMP1 to promote the proliferation of SP18 cells.LMP1 may improve the cell proliferation index and induce the proliferation of SP18 cells through TRADD.

To explore the relationships between traditional Chinese medicine (TCM) constitutional types and overweight or obesity so as to provide evidence for adjusting constitutional bias and preventing and treating obesity.

AIM: To construct pVAX1-GrB. METHODS: Lymphocytes from human laryngeal carcinoma tissue were separated from tumor tissue. The fragment of granzyme B (GrB) was amplified by RT-PCR and was recombined to the downstream of T7 promoter in the vector pVAX1. The construction was transfected into Hep2 cells with lipofectamine 2000. The expression of protein was identified by indirect immunofluorescent antibody assay. RESULTS: It has been proved that the sequence of the RT-PCR product was totally consistent with the data of GenBank by DNA sequencing analysis. The GrB cDNA fragment was cloned into the vector of pVAX1 in the right direction and the open reading fragment of GrB was maintained. The target protein was detected in the transfected Hep2 cells. CONCLUSION: The pVAX1-GrB plasmid was successfully constructed and expressed. [

Objective To investigate the current status and awareness of pharmaceutical services in hospitals in Guizhou province and to provide a reference for exploring and carrying out pharmaceutical service fees.Methods The questionnaire was designed by the"wjx.cn"website.Three kinds of questionnaires were designed for pharmacists,doctors,nurses,and patients as the research objects,with corresponding differences in some questions,and promoted on WeChat,Dingxiangyuan,and other network platforms.Results A total of 655 questionnaires were collected,and 639 valid questionnaires were recovered,with an effective recovery rate of 97.56%.324 pharmacists(50.70%),82 doctors and nurses(12.83%),233 patients(36.46%)were surveyed.The average approval score of these three groups of respondents on pharmaceutical service fees was 4.67,4.23,and 4.22,respectively(full score:5).Conclusions Overall,pharmacists'professional services have received support from medical staff and patients.However,patients'pharmaceutical service projects currently focus on dispensing services.The recognition of pharmacists'work and the public's awareness of pharmaceutical services can be improved by enhancing the professional ability of pharmacists,strengthening publicity and guidance,and exploring"Internet+pharmaceutical services",etc.,to promote the sustainable development of pharmaceutical services.

OBJECTIVE：To establish the metho d for the concentration det ermination of foretinib derivative LWK- 126 in liver microsomes，and to study its metabolism stability in liver microsomes of rats ，Beagle dogs and human. METHODS ：In the in vitro incubation system of liver microsomes ，LWK-126 was dissolved in liver microsomal incubation systems of rats ，Beagle dog and human initiated by reduced nicotinamide adenine dinucleotide phosphate solution. After incubation in water at 37 ℃ for 0，5，10，20， 30 and 60 min，the reaction was terminated with acetonitrile containing internal standard （1 μg/mL tolbutamide）. UPLC-MS/MS method was applied to determine the concentration of LWK- 126 in the incubation systems. The determination was performed on Waters BEH C 18 column with mobile phase consisted of water （containing 0.1％ formic acid ）-acetonitrile（containing 0.1％ formic acid）by gradient elution at the flow rate of 0.4 mL/min. The column temperature was 30 ℃，and the sample size was 2 μL. The mass spectral analysis was performed in a positive electrospray ionization mode ，and the full MS experiment was run with the selective reaction monitoring mode with a scanning range of m/z 50→1 200. Taking the concentration of LWK- 126 at 0 min as reference，the remaining percentage and the enzyme kinetic parameters were calculated. RESULTS ：The linear range of LWK- 126 was 0.05-15 μg/mL（R 2＝0.999 2）；the lower limit of quantification was 0.05 μg/mL，and the lowest detection limit was 0.01 μg/mL. The precision，accuracy，extraction recovery and matrix effect all met the analysis requirements of biological samples. The remaining percentage of LWK- 126 in liver microsomes of human ，rats and Beagle dogs for 60 min were （33.17±4.52）％，（3.14± 6.73）％，（1.38±5.85）％；t1/2 of them were 33.15，11.76，5.62 min；the clearance rates were 38.45，118.81，245.76 μL（/ min·mg）， respectively. CONCLUSIONS ：The method for the content ； determination of LWK- 126 in liver microsomes is established successfully. The order of metabolic stability of LWK- 126 in 〔2016〕4015） liver microsomes of different species is human ＞rats＞Beagle dogs.

Primary acinic cell carcinoma (ACC) of the lung is extremely rare. The World Health Organization tumor classification defines ACC as "a malignant epithelial neoplasm that demonstrates some cytological differentiation towards (serous) acinar cells". It is considered to be a low-grade malignant tumor. Since the first case described by Fechner in 1972, less than 30 cases have been reported in the literature. The rarity of this tumor may leads it to be confused with other primary lung tumors and incorrectly diagnosed. We reported a female patient with primary ACC of the lung with mediastinum deviation at age of 27 years received a right pneumonectomy. She was followed up for 12 months postoperatively and remains well.