Objective:A novel tuberculosis DNA vaccine integrating siRNA,fusional antigen and hIL-12 triple expression units was constructed in our laboratory.Methods:To evaluate the independent expression of the three expression units,two eukaryotic expression plasmid pVAX1-siRNA-PVAE[EGFP]-hIL12 with TB fusional gene Ag85B-ESAT6(PVAE) and enhanced green fluorescent protein(EGFP),and pVAX1-siEGFP-PVAE[EGFP]-hIL12 with siRNA coding sequence targeting EGFP instead of Mcl-1L were constructed.Then two plasmids were used to transfer human embryonic kidney 293 cells.Based on EGFP report gene,it was demonstrated that siRNA expression unit and fusional antigen gene were independently expressed.Results:The hIL12 expression at 48h and 72h post transfection were also detected by ELISA analysis up to 1571.63pg/ml and 2392.25pg/ml respectively in the cell culture fluid.Conclusion:The results demonstrated that the novel DNA vaccine with siRNA,TB fusional antigen and hIL12 three expression units in the same plasmid frame is successfully constructed and independently expressed in eukaryotic cells.It laid a foundation for further animal model study on anti-tuberculosis effects of this novel DNA vaccine.

Objective To improve the understanding and diagnosis of primary hypertrophic os-teoathropathy(PHO).Methods A case of PHO was reported.The clinical data and the process of the diagno-sis and treatment was analyzed retrospectively,and the related literature were reviewed.Results The patient was a young man without family history of PHO.He had symptoms since age 16.He had clubbing fingers and toes,hypertrophic skin,joint swelling,hyperhidrosis and radiographic evidence of periostitis.Thus the disease was diagnosed as PHO.The patient was treated with NSAIDs and the symptoms relieved very quickly.Conclusion Radiographic examination should be taken in time when young males have the general characters of clubbing fingers and toes,hypertrophic skin changes.If the periostitis presents,the final diagnosis of PHO can be confirmed.

Objective To investigate the application of Critical Care Chest Ultrasonic Examination (CCUE)-plus (CCUE-plus) in the etiological diagnosis in patieuts with dyspnea and/or hemodynamic instability caused by abdominal abnormalities.Methods Patients who suffered from dyspnea and/or hemodynamic instability in the Department of Critical Care Medicine,Xiangya Hospital,Central South University from September 2013 to September 2016 were recruited in this study.A total of 255 consecutive patients completed CCUE within 2hrs of admission.If the diaphragm could not be seen in the routine phrenic points according to Bedside Lung Ultrasound Evaluation (BLUE) protocol,it would be found along midaxillary line and defined m-point.The 59 patients with altered diaphragmatic position (m-point was more than 2 cm higher than phrenic point) received sequential abdominal ultrasonography.The latter ultrasonographic findings were compared with CT results.Results There were 42 (71.19％) cases with positive findings of abdominal ultrasonography,including 18 cases of seroperitoneum,16 cases of intestinal obstruction and 8 cases combined.Compared with 56 patients who applied with CT exam,the abdominal ultrasonography revealed a sensitivity of 76.7％ and a specificity of 100.0％ to diagnose seroperitoneum (AUCRoc 0.917);whereas the sensitivity was 75.0％ and the specificity was 90.9％ (AUCRoc 0.778) to diagnose intestinal obstruction.Moreover,there were 44 (74.58％) patients with normal left ventricular systolic function;more than three quarters (46/59,77.97％) patients had pulmonary consolidation.Conclusion In patients with dyspnea and/or hemodynamic instability caused by abdominal abnormalities and altered diaphragmatic position in BLUE protocol,CCUE-plus protocol has a high positive predictive value of more than 90％ in abdominal abnormality.The findings of abdominal ultrasonography may change therapeutic target from cardio-pulmonary optimization to relief of intestinal obstruction or drainage of seroperitoneum.

Objective To further control the indications and to improve the effect of intra-esophageal metallic stents.Methods 83 metallic stents were be planted into esophageal through endoscopy and under X-ray TV guiding using two kinds of stent meterials in 72 cases with benign or maligant stenosis(esophageal,cardiac orifice).Results The operation of planting stents was successful in all patients and the effect of treatment of esophageal stenosis with stents covered by dacron kint membrane was best.The clinical symptoms and food intaking ablities of all patients were improved.Conclusion Home-made nitinol stents can be plant in treatment of malighent esophageal stenosis.It should be first choice for the patients without indications of surgery or after operation.

Objective To validate a new standardized training program for urological surgeons to improve their laparoscopic surgical skills. Methods The laparoscopic surgical training program was carried out by using the traditional mechanical simulators and animal models. Thirty-three trainees participated the urological laparoscopic surgical training program. Initially, the novices were assigned to practise basic laparoscopic skills step by step on the simulator with fixed trocar positions. After a period of basic training, they were allowed to practise on animal models for some particular proce-dures. Results All trainees (33/33, 100.0%) participated were able to perform all basic techniques skillfully and completed laparoscopic anastomosis accurately after the training. The time required for performing the partial nephrectomy, dismembered pyeloplasty and ureteral reimplantation on animal models declined from 64.0±18.4, 127.54±17.5 and 75.84±11.6 min at the beginning to 30.94±3.8, 65.2±7. 5 and 37.7±7.2 min after practicing these procedures 8 times (P<0.01). They could un-derstand the crucial procedures of the laparoseopic surgeries after 6 to 8 special trainings on animal models. Fifteen trainees (15/33, 45.5%) had started to carry out laparoscopic surgeries after-finish- ing the training program. Conclusions Our program enables the participants to improve their techniques in complicated laparoscopic surgeries. The challenging parts of reconstructive laparoscopic surgeries such as laparoscopic dismembered pyeloplasty can be taught by using animal models. This program could be incorpo-rated easily by all urological departments developing a laparoscopie surgical training program.

Objective To improve the level of CT in diagnosing LDH and to provide the proof for selecting method of clinical treatment in lumbar disc herniation (LDH). Methods 218 cases of LDH showed by CT and operative exploration were analysed retrospectively.Results The accurate rats of CT diagnosis of LDH were 95.4%. According to CT findings it may be divided into 5 types: Ⅰ was central type, Ⅱ was postero-lateral type, Ⅲ was forminal type, Ⅳ was extreme lateral type, was nucleus pulposus type. Conclusion CT plays an important role in diagnosis and selecting methods of clinical treatment of LDH; It is signficance to enhance the accurate rats of CT diagnosis and the effect of treatment by distinguish nucleus pulposus and fibrous scar tissue from CT finding of LDH.

Objective To conclude and compare the technical characteristics and clinical efficacy of retroperitoneal and transperitoneal approach for robotic partial nephrectomy.Methods 418 patients received robotic partial nephrectomy conducted by one surgeon in the urology department of PLA General Hospital from 2013, December to 2015, October, including 157 cases in retroperitoneal approach and 261 cases in transperitoneal approach.There were no significant difference between the transperitoeal and retroperitoneal approach in patient age(51 years, 51 years, P=0.593), BMI(25.5 kg/m2,25.6 kg/m2, P=0.331), gender(male/female:111/46,186/75,P=0.184), location(left/right,80/77,123/138,P=0.575), tumor size(3.1cm,3.5cm,P =0.356), comorbidities, proportion of solitary kidney(11.5%, 9.6%,P=0.253) or bilateral tumors(1.3%,3.4%,P=0.179), RENAL score(6(5-8),6(5-8),P=0.083), ASA score(2 vs.2)(P=0.310) and preoperative serum creatinine(74.7 μmol/L,76.7 μmol/L, P=0.736).Patients in the retroperitoeal approach group were more likely to have abdomen surgery history (21.7%,14.2%,P=0.049).there was significant difference between two approaches in tumor location in the kidney ( anterior/posterior/others: 9/140/8, 212/36/13, P <0.001 ) Results The operation time (105min, 115min, P =0.041 ) and warm ischemia time ( 15min, 20min, P <0.001 ) were shorter and estimated blood loss(50ml,75ml,P<0.001) was less in the retroperitoneal group.No significant difference was observed in postoperative 24h blood loss(45ml,50ml,P =0.093), intraoperative transfusion rate (2.5%,6.5%,P=0.072) and collective system injury(24.8%,27.6%,P =0.539) between the two groups.Peritoneum break occurred in 25 cases in retroperitoneal group, which were repaired by Hem-o-lock.One intestinal injury occurred in transperitoneal group and was repaired intraoperatively.Three and nine cases in the retroperitoneal and transperitoneal group were converted to radical nephrectomy.One case in the transperitoneal group was found positive margin and received laparoscopic radical nephrectomy.Two liver injury and five spleen injury occurred in the transperitoneal group.For postoperative food intake(1 day, 2 days, P <0.001 ) , the length of indwelling drainage ( 3 days, 4 days, P <0.001 ) and the length of postoperative stay ( 5 days, 6 days, P =0.001 ) , the retroperitoneal group had one day shorter than the transperitoneal group.With the median following-up time of both group, 11 months (IQR:6-16) in RPRPN group and 12 months ( IQR: 7-19 ) , no subject with a tumor recovery or metastasis.Conclusions Retroperitoneal robotic partial nephrectomy is feasible and effective, which is similar to transperitoneal approach.The retroperitoneal approach is superior to transperitoneal approach for posterior, lateral and superior renal tumors.

ObjectiveTo investigate the alteration of plasma C-reactive protein (CRP) count in patients with non-valvula atrial fibrillation combining thrombosis.MethodsA total of 154 patients with non-valvula atrial fibrillation were divided into thrombus group (n = 46) and non-thrombus group (n = 108)in accordance with transesophageal echocardiography (TEE) results.The concentration of CRP by scattering turbidimetry, D-dimer by immunoturbidmetry, left atrium diameter (LAd), fraction shortening (FS) and left ventricular ejection fraction (LVEF) by TEE or echocardiography were detected.Logistic multi-factors regression analysis was performed.ResultsThere were significant differences in CRP [(5.77 ± 6.37)mg/L vs (1.73 ±2.39)mg/L,P =0.003], LAd [(59.86 ± 10.70) mm vs (47.97 ± 13.19)mm,P ＜0.001]and LVEF[(58.75 ±8.28)％vs (64.10 ±6.75)％, P ＜0.001]between thrombus group and nonthrombus group.The results of logistic regression analysis found CRP ≥ 1.33 mg/L(OR 2.856, 95％ CI 1.235-6.600,P =0.014) and LAd≥54.5 mm(OR 4.236, 95％ CI 1.877-9.562,P =0.001) were independent risk factors of patients with non-valvula atrial fibrillation combining with thrombosis.ConclusionsCRP and LAd are independent risk factors for patients with non-valvula atrial fibrillation combining with thrombosis.Inflammation may involve with the formation of thrombosis.

BACKGROUND: Vascular endothelial growth factor (VEGF) and its receptor fms-like tyrosine kinase-1 (Flt-1) have a very important position in the field of male reproduction. However, it is still unclear about their expression meaning and regulatory mechanism in the reproductive system. OBJECTIVE: To study the expression and location of VEGF and Flt-1 in the epididymal sperm of adolescent male rats. METHODS: The expression of VEGF and Flt-1 was detected in 10 adolescent SD rats. The concentration of the sperm was (30-40)×109/L. Immunofluorescent staining was used for VEGF and Flt-1 expression and location in the sperm. RESULTS AND CONCLUSION: Immunofluorescent staining showed that VEGF and Flt-1 were both localized in the acrosome of sperm head, as well as in the neck, middle and principal segment of sperm tail. Specific expression patterns of VEGF and Flt-1 in the sperm of rats display that they may participate in spermiotelcosis, relevant to movement, capacitation and acrosome reaction of the sperm.

In this study, we investigated the effect of Cigu Xiaozhi formula on HSC-T6 activity in hypoxic microenvironment based on network pharmacology and computer-aided drug design, and predicted and verified its possible targets and related signaling pathways. The potential active components and targets of Cigu Xiaozhi formula were screened by searching Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP), Encyclopaedia of Traditional Chinese Medicine (ETCM) and Bioinformatics Analysis Tool for Molecular Mechanism of Traditional Chinese Medicine (BATMAN-TCM) databases, and the liver fibrosis related targets retrieved from Gene Cards and Pharm GK database were integrated to obtain the potential targets of Cigu Xiaozhi formula in the treatment of liver fibrosis. GO enrichment analysis and KEGG signaling pathway enrichment analysis were performed on Omic Share platform, and Cytoscape software was used to construct the "potential active ingredient-key target-pathway" network. The active components and target proteins were subjected to molecular docking analysis by Auto Dock software. According to the results of molecular dynamics simulation and binding free energy calculation, the top 5 active components with degree were scored. The active components stigmasterol and β-sitosterol were subjected to molecular docking. CoCl₂ was used to induce HSC-T6 cells to construct hypoxia model in vitro. The cell viability was detected by CCK-8 assay, and the optimal time and concentration of hypoxia model of HSC-T6 cells was determined to be 100 µmol·L^-1 CoCl₂ for 24 h. Under hypoxia condition, HSC-T6 cells were activated, the wound healing rate was significantly increased, and the fluorescence signal of activation marker protein α-smooth muscle actin (α-SMA) was significantly enhanced. However, 6% drug-containing serum could inhibit the activation of HSC-T6 cells, and the wound healing rate was significantly decreased, and the fluorescence signal of α-SMA was significantly weakened. Further studies showed that the expressions of hypoxia-inducible factor-1α (HIF-1α), α-SMA and key proteins of Hedgehog (Hh) signaling pathway in HSC-T6 cells were up-regulated under hypoxia, while the expressions of HIF-1α, α-SMA, Patched-1 (Ptch-1) and glioma related oncogene homology-1 (Gli-1) were down-regulated in 6% drug-containing serum group, the YC-1 group and the cyclopamine group. These results indicated that HIF-1α and Hh signaling pathways were involved in the activation of HSC-T6 cells, and the traditional Chinese medicine Cigu Xiaozhi formula could inhibit the activation of HSC-T6 cells, and the mechanism may be related to the inhibition of HIF-1α expression and the blocking of Hh signaling pathway. In conclusion, Cigu Xiaozhi formula can inhibit the activation of HSC-T6 cells by directly acting on HIF-1α and Hh signaling pathway, and exert an anti-hepatic fibrosis effect. The animal experimental protocol has been reviewed and approved by Laboratory Animal Ethics Committee of Gansu University of Chinese Medicine, in compliance with the Institutional Animal Care Guidelines.