Objective:To investigate the immunogenicity of pORF5 plasmid protein,and further to screen and identify its im-munodominant domian.Methods: 10 different fragments of pORF5 gene including full length were amplified from the DNA of Chlamydia trachomatis serovar D by PCR and cloned into appropriate site of pGEX-6p vector to construct recombinant vectors after digested with BamHⅠand NotⅠrestriction endonucleases.After identification by PCR and sequencing,the recombinant plasmids were transformed into XL1 Blue E.coli to express the GST fusion proteins.ELISA and Western blot were carried out to identify the immunogenicity and immunoreaction of pORF5 plasmid protein.10 different fragments were reacted with sera from patients urogenitally infected with Chlamydia trachomatis, mouse polyclonal antibodies and mouse monoclonal antibodies of pORF5 plasmid protein with ELISA method.Results: pORF5 plasmid protein displayed strong immunogenicity and could induce a strong antibody response in human.The reactivity of human antibodies almost completely disappeared,when the native structure of pORF5 plasmid protein was de-stroyed.F6 that only lacked the N-terminal 66 amino acids was recognized by antibodies in ELISA as strongly as the whole pORF5 plasmid protein was.However,no other fragments were significantly recognized although there was a minimal reactivity of F2 and F3 with antibodies.Conclusion:pORF5 plasmid protein was an immunodominant antigen containing conformation-dependent epitope,and the C-terminal three quarters of pORF5 amino acid sequence was required for maintaining its immune dominance and conformation.The significance of the above findings lay a foundation for the further study on pORF5 protein function and vaccine development.

Objective To investigate the expression of Oct4 in liver cancer, and the interrelation of the Oct4 and Wnt/β-catenin genes in hepatocellular carcinoma( HCC) cell line HepG2. Methods RTPCR technique was used to detect the expression of Oct4 and β-Catenin in HCC specimens; RNAi was used to knock-down the expression of Oct4 in HepG2, and the change of Wnt/β-catenin related genes were detected by Real time-PCR. Results In HCC specimens, the expression of Oct4 and β-Catenin in tumor and cirrhotic liver tissues were stronger than normal liver tissues. In SiRNA Oct4 HepG2 cells, the expression of Oct4 was downregulated, and β-catenin as well as Wnt10b were in a positive correlation with Oct4, TCF3 was in negative correlation with Oct4. Clone formation and move ability of the HepG2 were downregulated. Conclusions The expression of Oct4 was higher in tumor tissues than in normal liver tissues. Silencing Oct4 by SiRNA-0ct4 in HepG2 resulted in decreased ability of clone formation and cell movement.

Objective To investigate the variation characteristics of adenovirus type 55 (HAdV-B55) gene on plateaus.Methods Throat swabs were collected from HAdV-B55 infected patients and used for virus isolation in HEp-2 cells.The whole-genome sequence was obtained by PCR and sequencing.HAdV-B55 gene sequence was blast with the previously reported virus.Results HAdV-B55 strains were isolated from throat swabs, which were named LS89/Tibet/2016.The whole-genome sequence was obtained and submitted to GenBank with the accession number of KY002683.No large fragment gene recombination was found between this HAdV-B55 strain and previous strains, and the sequence similarity with QS-DLL strain was 99.9%.Conclusion This study provides more information for the evolution patterns of adenovirus 55 and will contribute to the prevention and control of HAdV-B55 infection in the future.

Objective To evaluate the effectiveness of free anterolateral thigh flaps for the reconstruction of lower extremities vast soft tissue defect caused by Gustilo Ⅲ open fracture. Methods From June,2013 to December, 2016,pedicled vascular anterolateral thigh flaps with end-to-side vascular anastomosis were adopted to repair 10 cas-es of lower legs caused by Gustilo Ⅲ open fracture and soft tissue defects. Among those cases,6 cases combined with anterior tibial arteries injury and 4 cases with posterior tibial arteries injury. The receiving arteries were anastomosed with flaps arteries in end-to-side method and the receiving veins were anastomosed in end-to-end method. All pa-tients were regular follow-up, and follow-up contents included flap appearance, color, limb ischemia. Results All the flaps survived in10 cases. Venous crisis was happened in one case,and flap survived after the exploration. Two cases delayed healing because of wound infection.All the 10 cases were followed up for 3-24 months,with an average of 10 months.The skin flaps of 10 cases were thin and well fitted,with good appearance and soft texture. The color of the flaps were similar to that of the recipient areas.The appearance and function of the affected limbs were satisfacto-ry. Conclusion Free anterolateral thigh flap with end-to-side vascular anastomosis is an ideal method to repair the vast soft tissue defects caused by Gustilo Ⅲ open fracture.

Objective: To investigate the clinical effect of one-stage revision combined with intra-articular injection of antifungal agents in the treatment of chronic periprosthetic fungal infection. Methods: A retrospective analysis of 11 patients(4 hips, 7 knees) admitted with chronic periprosthetic fungal infection at Department of Orthopaedics, First Affiliated Hospital of Xinjiang Medical University from January 2004 to April 2016.There were males and females with an age of 67 years (range:47-77 years). Each patient underwent single-stage revision including aggressive soft-tissue debridement. Liquid samples and tissue samples were immediately sent to the microbiology laboratory for drug sensitivity testing and histological analysis. Removed the infected components and cement thoroughly, pouring powdered vancomycin into the medullary cavity and direct intra-articular injection of fungussensitive antibiotics. The patients with infected hips received an uncemented prosthesis and 0.5 g of gentamicin loaded commercial cement was received by the patients with infected knee.After that, a new prosthesis was implanted.Long-term combination therapy of antibacterial agents and antifungal agents were given after operation. Recurrence of infection and clinical outcomes were evaluated. The follow-up period was 5 years (range: 2-12 years). Results: One patient died of acute heart failure on the eighth postoperative day.Three infection cases were recurred.Eight cases had satisfactory outcomes and required no additional surgical or medical treatment for recurrence of infection. The Harris hip score assessed preoperatively and at latest follow-up was increased from 39.25±5.12 to 79.50±4.79, the difference was statistically significant (t=-11.356, P=0.001).The Hospital for Special Surgery knee score was improved from preoperative 46.25±5.61 to final follow-up 80.50±5.06, and the difference was statistically significant (t=-9.930, P=0.002). Conclusion Treatment of chronic fungal periprosthetic joint infection with single-stage revision can be fairly effective for achieving acceptable functional outcomes.

Objective:To investigate the effects of SuperPATH approach versus conventional posterolateral approach in total hip replacement on inflammatory response, hip function, and quality of life in patients with hip diseases. Methods:The clinical data of 140 patients with hip diseases who underwent total hip replacement in Shanxian Central Hospital from March 2017 to May 2019 were retrospectively analyzed. These patients were divided into SuperPATH approach ( n = 70) and posterolateral approach ( n = 70) groups. Operation-related indexes, inflammatory response indexes, hip function, quality of life, and pain were compared between the two groups. Results:Intraoperative blood loss was significantly less in the SuperPATH approach group than in the posterolateral approach group [(105.40 ± 15.11) mL vs. (196.89 ± 24.26) mL, t = 26.74, P < 0.001]. Incision length, postoperative time to getting out of bed, length of hospital stay in the SuperPATH approach group were (6.85 ± 1.42) cm, (2.92 ± 0.28) days, and (6.67 ± 1.36) days, respectively, which were significantly shorter than those in the posterolateral approach group [(13.07 ± 1.89) cm, (8.36 ± 1.45) days, (10.91 ± 1.34) days, t = 19.36, 30.82, 18.58, P < 0.001]. Operative time was significantly longer in the SuperPATH approach group than in the posterolateral approach group [(69.38 ± 8.62) minutes vs. (60.45 ± 7.79) minutes, t = 6.43, P < 0.001). The scores of social role functioning, general health perceptions, vitality, mental health, bodily pain, emotional role functioning, physical functioning, and physical functioning measured 6 months after surgery were significantly higher in the SuperPATH approach group than in the posterolateral approach group ( t = 9.12, 11.80, 11.64, 11.69, 6.45, 11.79, 6.04, 10.74, all P < 0.001). There were no significant differences in C-reactive protein and erythrocyte sedimentation rate measured 3 and 14 days after surgery between the two groups (both P > 0.05). Harris score used for evaluation of hip function 1 month after surgery was significantly higher in the SuperPATH approach group than in the posterolateral approach group [(76.42 ± 4.17) points vs. (69.37 ± 5.11) points, t = 8.94, P < 0.001]. The Visual Analog Scale score 3 days after surgery was significantly lower in the SuperPATH approach group than in the posterolateral approach group [(3.18 ± 0.21) points vs. (4.26 ± 0.29) points, t = 25.23, P < 0.001]. Conclusion:Compared with the conventional posterolateral approach, the SuperPATH approach for total hip arthroplasty takes longer operative time, but it can better reduce early postoperative pain, promote hip function recovery, and improve quality of life.

Objective:To evaluate the clinical effects of anatomical reconstruction of the deltoid ligament and medial migration calcaneal osteotomy in the treatment of chronic ankle instability after trauma.Methods:A retrospective study was conducted to analyze of the clinical data of 16 patients with chronic post-traumatic instability of the medial malleolus who had been treated at The Eighth Department of Orthopedics, Foshan Hospital of Traditional Chinese Medicine from December 2015 to February 2017. There were 14 males and 2 females with an age of (28.1±4.2) years. Seven left sides and 9 right sides were affected; the time from injury to operation was (8.9±2.4) months. Before operation, X-rays (anteroposterior, lateral and Saltzman views) and MRI of weight-bearing ankle were taken. All patients were treated by anatomical reconstruction of the deltoid ligament using the autologous semitendinosus and medial migration osteotomy of the calcaneus. The time for injury healing and occurrence of complications were recorded. The talus tilt angle, Meary angle, hindfoot valgus angle, visual analogue scale (VAS), and the American Orthopedic Foot & Ankle Society (AOFAS) ankle-hindfoot function score were compared between the preoperation and the last follow-up. The Sefton evaluation for efficacy in ankle ligament reconstruction was used to assess stability of the ankle joint.Results:Of this cohort, 14 patients were followed up for (16.4±4.9) months after operation and 2 patients lost to follow-up. The 14 patients all returned to normal physical activities 3 months after operation. All incisions healed at the first stage with no infection. One patient experienced pain at the site for harvest of the semitendinosus but the symptoms were relieved after rehabilitation treatment like massage and physical therapy. At the last follow-up, the talus tilt angle [1.0 (0.0, 2.0)°], Meary angle (1.4°±4.2°), hindfoot valgus angle (3.2°±2.4°), VAS score [0.5 (0.0, 1.0) points], and AOFAS ankle-hindfoot score [(89.2±6.1) points] were all significantly improved compared with the preoperative values [8.3°±1.8°, 0.8°±3.8°, 9.9°±3.4°, (5.7±2.5) points, and (49.6±9.8) points] (all P<0.05). According to the Sefton evaluation, the stability of the ankle joint was excellent in 9 cases, good in 4 cases, and fair in 1 case. Conclusion:In the treatment of chronic ankle instability after trauma, anatomical reconstruction of the deltoid ligament and medial migration calcaneal osteotomy is safe and effective, resulting in limited complications.

Objective:To explore the application effect of flipped classroom combined with online interactive teaching in clinical clerkship of cardiology.Methods:The study collected 56 students from Batch 2017 five-year clinical medicine undergraduate class of Medical College of China Three Gorges University and they were randomly divided into experimental group and control group in average. The control group adopted the traditional teaching method while the experimental group adopted the combined teaching method. The differences of test scores after teaching were compared between the two groups and the feedback of students on the two methods was also analyzed. SPSS 12.0 was used for t test and chi-square test. Results:The final score of clinical thinking in the experimental group (84.38±3.18) was significantly higher than in the control group (75.43±5.85) ( P<0.05), and the satisfaction of the experimental group to the teaching mode was [86% (24/28)], which was significantly higher than that of the control group [50% (14/28)]. More than 80% of the students in the experimental group regarded that the combined teaching method was helpful to promote the interaction between teachers and students, master relevant knowledge, and improve learning interest and learning efficiency. Conclusion:This combined teaching method can enhance students' clinical thinking ability, learning enthusiasm and efficiency, and finally effectively improve the effect of clerkship.

BACKGROUND/OBJECTIVES: The study examined the association between homocysteine and diabetes mellitus in patients with H-type hypertension and assessed the possible effect modifiers. SUBJECTS/METHODS: This cross-sectional study included 1,255 eligible participants in the ‘H-type Hypertension Management and Stroke Prevention Strategic International Science and Technology Innovation Cooperation Project’ among rural Chinese people with H-type hypertension. A multivariate logistic regression model was used to evaluate the relationship between homocysteine and diabetes mellitus. RESULTS: The mean level of total homocysteine (tHcy) in the diabetes mellitus population was 19.37 μmol/L, which was significantly higher than the non-diabetic patients (18.18 μmol/L). When tHcy was analyzed as a continuous variable, the odds ratio (OR) of diabetes was 1.17 (95% confidence interval [CI], 1.01–1.35; per interquartile range). When tHcy was stratified according to the quintile, the ORs for diabetes were 2.86 (95% CI, 1.22–6.69) in the highest quintile (tHcy ≥ 20.60 μmol/L) compared to the reference group (tHcy < 12.04 μmol/L). When tHcy was grouped by 15 μmol/L and 20 μmol/L, patients with tHcy ≥ 20 μmol/L had a significantly (P = 0.037) higher risk of diabetes (OR, 2.03; 95% CI, 1.04–3.96) than in those with tHcy < 15 μmol/L. Subgroup analysis showed that the tHcy-diabetes association was unaffected by other variables. CONCLUSION In this study of rural Chinese people with H-type hypertension, the tHcy levels showed a positive association with diabetes mellitus. This independent association is unaffected by other potential risk factors.

BACKGROUND/OBJECTIVES: The study examined the association between homocysteine and diabetes mellitus in patients with H-type hypertension and assessed the possible effect modifiers. SUBJECTS/METHODS: This cross-sectional study included 1,255 eligible participants in the ‘H-type Hypertension Management and Stroke Prevention Strategic International Science and Technology Innovation Cooperation Project’ among rural Chinese people with H-type hypertension. A multivariate logistic regression model was used to evaluate the relationship between homocysteine and diabetes mellitus. RESULTS: The mean level of total homocysteine (tHcy) in the diabetes mellitus population was 19.37 μmol/L, which was significantly higher than the non-diabetic patients (18.18 μmol/L). When tHcy was analyzed as a continuous variable, the odds ratio (OR) of diabetes was 1.17 (95% confidence interval [CI], 1.01–1.35; per interquartile range). When tHcy was stratified according to the quintile, the ORs for diabetes were 2.86 (95% CI, 1.22–6.69) in the highest quintile (tHcy ≥ 20.60 μmol/L) compared to the reference group (tHcy < 12.04 μmol/L). When tHcy was grouped by 15 μmol/L and 20 μmol/L, patients with tHcy ≥ 20 μmol/L had a significantly (P = 0.037) higher risk of diabetes (OR, 2.03; 95% CI, 1.04–3.96) than in those with tHcy < 15 μmol/L. Subgroup analysis showed that the tHcy-diabetes association was unaffected by other variables. CONCLUSION In this study of rural Chinese people with H-type hypertension, the tHcy levels showed a positive association with diabetes mellitus. This independent association is unaffected by other potential risk factors.