Objective To construct Cox7a2 fluorescent vector and study its effect on cytochrome C oxidase (COX) activity in mouse Sertoli cell line TM4. Methods The coding region of Cox7a2 was amplified from mouse Sertoli cell line TM4 by RT-PCR. The PCR product was inserted into pEYFP-C1 vector with BamH I and EcoR I restriction site, and confirmed by DNA sequencing. The recombinant fusion protein vector was amplified by transforming into DH5a and transfected into TM4 cells. The protein expression was identified by Western blot. COX activity was measured by spectrophotometer 6, 12, 24 and 48 h after the transfection of recombinant vector into the TM4 cell line. Results The entire coding sequence of Cox7a2 was cloned with 252 bp length. Plasmid pEYFP-C1-Cox7a2 vector was constructed and the positive clones were verified by restriction enzymes digestion and DNA sequencing. The transfection efficiency of the TM4 cell line was about 70% and 37000 D fusion protein was obtained. The COX activities were (0.642±0.051), (0.542±0.049), (0.311±0.021) and (0.216±0.010) U/mg 6, 12, 24 and 48 h after the transfection of recombinant vector in the TM4 cell line. Meanwhile, the COX activities were (0.714±0.064) and (0.653±0.031) U/mg in non-tranfected and naked vector group respectively. Compared with the non-tranfected group, COX activity decreased significantly 12, 24 and 48 h after the transfection. Conclusions The recombint plasmid vector was successfully constructed. Cox7a2 gene has an inhibiting effect on COX activity and may play an important role in the regulation of COX activity in mouse Sertoli cell line TM4.

Objective To investigate clinical efficacy of transforaminal approach debridement with fusion,thoracolumbar single segment of Brucella spondylitis pedicle screw fixation (TLIF surgery).Methods We analyzed retrospectively the clinical data of 28 patients with Brucella spondylitis treated in our department between January 2009 and January 2014 with TLIF surgery (Group A) and internal fixation with a simple posterior anterior interbody disease debridement,autogenous bone graft (Group B).The two groups were compared in operation time,blood loss,postoperative ambulation time,hospitalization days,erythrocyte sedimentation rate (ESR),Creactive protein (CRP),American Spinal Injury Association (ASIA) classification,visual analogue scale (VAS),Oswestry Disability Index (ODI),Cobb angle of vertebral bone graft healing,and complications.Results All the patients were followed up for an average of 20.2 months (18 to 27 months).They were all cured.Compared with those in Group B,patients in Group A had shorter operation time (164.60±59.19)min,significantly reduced blood loss (346.00±108.90)mL and complications (1 case);significantly shorter postoperative ambulation time (3.36±1.11 days),hospitalization days (17.36 ± 4.19) days and duration (13.16 ± 3.94) months (P ＜ 0.05).The two groups did not significantly differ in VAS scores,ODI,ESR CRP,or Cobb angle (P＞0.05).Conclusion On the basis of norms of anti-drug treatment for brucellosis,TLIF surgery on Brucella spondylitis has the advantages including less trauma,shorter operation time,easier operation,less bleeding,earlier postoperative ambulation,and lower complication rate.

Objective: To analyze the clinical pathologic characteristics of cases with fluorescence in situ hybridization (FISH) positive of exfoliated urothelial cells, so as to evaluate the clinical utility of FISH in the diagnosis of urothelial carcinoma (UC).Methods: A total of 271 cases of FISH positive in Department of Urology of Peking University First Hospital from Apr.2012 to Sep.2015 were recruited in this study.Retrospective analysis was made on their clinical data.For FISH analysis, labeled probes specific for chromosomes 3, 7, 17, and the p16 (9p21) gene were used to assess chromosomal abnormalities indicative of malignancy.The positive predict values (PPV) of all the techniques were analyzed.Results: Of the 271 patients, 207 cases were UC, 7 cases were non-UC, and 57 cases were benign diseases.The PPV of FISH in detecting UC was 76.4%, while the 95% confidence interval (CI) 71.3% to 81.5%.In the cohort of FISH positive, this value was similar to that of urinary cytology (PPV 86.8%, 95% CI: 78.5%-95.0%).The PPV of FISH was lower than that of cystoscopy and ureteroscopy (PPV 96.1%, 95% CI: 91.7%-100.0%).There were significant differences between this study and the PPV of FISH reported abroad (PPV 53.9%, χ2=33.048, P<0.001).Of all the UC with FISH positive, bladder cancer showed an earlier pathological stage versus renal pelvic carcinoma and ureteral carcinoma, with significance (χ2=5.894, P=0.015, and χ2=13.601, P<0.001, respectively).However, no difference was found in the size, pathological stage and pathological grade of tumors between the urinary cytology positive group and the urinary cytology negative group.The rate of high-grade UC in ureteral carcinoma of FISH positive was 92.3%, much higher than that of ureteral carcinoma reported domestically.Conclusion: The PPV of FISH in detecting UC is higher relatively, with a better clinic value for Chinese patients.The ureteral carcinoma with FISH positive obtains a higher pathological grade, which is of great guiding significance for UC.

Objective To genotype mixed samples with next generation sequencing and evaluate its prospects in forensic DNA application. Methods Three mixed biological samples from rapes cases and their reference samples were collected. DNA was extracted using the MagAttract M48 DNA Manual Kit(200). The ForenSeqTMDNA Signature Prep Kit was used for library preparation, and next generation sequencing was performed on the MiSeq FGx system. The ForenSeqTMUniversal Analysis v1.2.1 software was used for data analysis. NGS-based STR results were compared with CE-based genotypes. Results A single length polymorphic STR allele in the mixed profile could be recognized as two sequence polymorphic STR alleles from different donors, which would assist mixed profile analysis. Such phenomenon was observed in D3S1358, D9S1122 and D13S317 in this work. Conclusion Our results suggested that precision STR genotyping of mixed samples based on NGS can provide more information and hints for mixed STR profile separation.

Objective: To understand characteristics of demographic, seasonal and spatial distribution of H5N1 cases in major countries of Asia (Indonesia, Cambodia, Vietnam, China) and Africa (Egypt). Methods: Through searching public data resource and published papers, we collected cases information in five countries from May 1st, 1997 to November 6th, 2017, including general characteristics, diagnosis, onset and exposure history, etc. Different characteristics of survived and death cases in different countries were described and χ² test was used to compare the differences among death cases and odds ratio (OR) and 95%CI value was used to compare death risk in different countries. Results: A total of 856 cases were reported in five countries with Egypt had the most cases (44.3%). The highest number of cases were reported in 2015 (18.3%). 53% cases were reported from January to March, and 96.1% of cases had the history of poultry exposure. 64.2% (43 cases) cases in China had live poultry market exposure, but the sick/dead poultry exposure was the major exposure for cases in other four countries. 452 death cases were reported in five countries, and the fatality rate was 52.8%. With Egypt as the reference group, the highest death risk was seen in Indonesia (OR (95%CI): 11.52 (7.46-17.77)), followed by Cambodia (OR (95%CI): 4.27(2.37-7.69)) and China (OR (95%CI): 2.87 (1.73-4.74)). The age distribution of death cases among 5 countries was statistically significant, and the highest fatality rate was in 15-54 years group in Egypt (83.6%, 102 cases), while in Cambodia the highest fatality rate was in 0-14 years group (76.9%, 30 cases). The highest number of deaths were reported in 2006, and 48.3% were reported from January to March. There was difference in exposure routes among 5 countries (χ²=43.85, P=0.001), 63.2% (24 cases) of the death cases in China had live poultry market exposure. 92.9% (79 cases), 83.3% (40 cases) and 100.0% (38 cases) death cases in Indonesia, Vietnam and Camodia had sick/dead poultry exposure, respectively;and 81.6% (31 cases) of the death cases in Egypt had backyard poultry exposure. Conclusion The geographical distribution, seasonal age, gender, exposure matter and outcome of H5N1 cases in five countries were different.

【Objective】 To investigate the current situation concerning volume control of red blood cells in additive solution produced by blood service in Chongqing, and to lay a foundation for promoting the homogenization of preparation process of red blood cells in additive solution. 【Methods】 A questionnaire was designed to investigate the factors related to the preparation of red blood cells in additive solution. The questionnaire was sent by Chongqing Association of Blood Transfusion via E-mail to 18 blood services in the city, and the collected data was sorted, revised and analyzed by research team. 【Results】 A total of 18 blood services(including 1 blood center + 1 sub-center, 6 central blood stations and 11 central blood banks) returned the questionnaires. The results showed that there were differences among blood services across Chongqing, regarding the centrifugal parameters during preparation, the operation mode and monitoring situation of the capacity control during preparation, and the formulation of the capacity standard of red blood cells in additive solution etc. 【Conclusion】 The preparation process of red blood cells in additive solution, produced by Chongqing blood services, should be further standardized, and the capacity control method of this product in Chongqing should be gradually unified to achieve regional homogeneity and to ensure blood safety.