Objective To construct the lentiviral vector to co-express enhanced green fluorescent protein (EGFP) gene and human insul in (insulin) gene, and to explore the condition to transfect human umbil ical cord mesenchymal stem cells (hUCMSCs) so as to lay a foundation for tissue engineered adipose reconstruction and transplantation in vivo infuture. Methods The insulin gene was cloned to lentiviral expression vector with EGFP [pLenti6.3-internal ribosome entrysite (IRES)-EGFP] by recombinant DNA technology, the positive clones were screened, and lentiviral packaged systems and target gene plasmid were co-transfected to package virus in 293T cells by lipofectin. The reporter gene expression was observed by fluorescent inverted phase contrast microscope, virus supernatant was collected, purificated and concentrated, and the titer of recombinant viruses was determinated. hUCMSCs from umbilical cord tissue of mature neonates were isolated and cultured by different multiple of infection (MOI, 0, 1, 3, 5, 7, 10, 15, and 20). By recombinant lentiviral infected hUCMSCs with reporter gene green fluorescent protein expression, the best MOI was screened; recombinant lentiviral infected hUCMSCs at the best MOI, then real-time PCR and Western blot methods were appl ied to detect insulin gene and insul in protein expression levels in cells. Results The recombinant lentiviral vector of co-expressing insulin gene and EGFP gene (pLenti6.3-insulin-IRESEGFP) was successfully constructed. Virus could be packaged, purificated and concentrated successfully. The virus titer was 1.3 × 108 TU/mL. The best MOI was 10 and the transfer efficiency was up to 90% in the same time. Real-time PCR results showed that insulin gene expression of transfected group was positive and non-transfected group was negative; Western blot detection confirmed that insul in protein expression of transfected group was positive in cells and supernatant, but that of non-transfected group was both negative. Conclusion Lentiviral vector pLenti6.3-insulin-IRES-EGFP carrying recombinant insulin gene could effectively transfect hUCMSCs and express insul in protein.
ObjectiveTo investigate the effectiveness of anterior cruciate ligament (ACL) reconstruction with the autologous ipsilateral peroneus longus tendon (PLT) under arthroscopy.MethodsA retrospective study was conducted on 35 patients with ACL rupture who underwent ACL reconstruction with autologous ipsilateral PLT under arthroscopy between October 2017 and October 2018. There were 19 males and 16 females with an average age of 43.4 years (range, 18-60 years), with 20 cases of left knee and 15 cases of right knee. The causes of injury included traffic accident in 14 cases, falling injury in 13 cases, and sports injury in 8 cases. The time from injury to operation ranged from 3 to 9 days (mean, 4.7 days). The patients suffered from swelling, pain, and limited mobility of knee joint before operation. The anterior drawer test, Lachman test, and pivot-shift test were positive before operation, whereas MRI was taken to confirm the ACL rupture. After operation, the patients were followed up every 3 months until the knee joint’s function returned to normal. MRI and X-ray films were used to observe the tendon-bone healing as well as the position of Endobutton suspension plate and hollow nail. The anterior drawer test, Lachman test, and pivot-shift test were conducted to observe the improvement of knee joint mobility. The functional improvements were evaluated by the International Knee Documents Committee (IKDC) score, Lysholm score, knee injury and osteoarthritis (KOOS) score.ResultsAll the 35 patients were followed up 12-18 months, with an average of 14.2 months. The incisions healed by first intention, and no complications such as infection, joint stiffness, and rerupture occurred. Postoperative anterior drawer test, Lachman test, and pivot-shift test turned to be negative of all patients. MRI showed that the ACL was continuous, and the tendon-bone in the distal femur tunnel and proximal tibia tunnel recovered well after operation. X-ray films showed that the positions of Endobutton suspension plate and hollow nail were stable. The IKDC, Lysholm, and KOOS scores at 3, 6, and 12 months after operation were significantly improved when compared with those before operation, and the scores were further improved with time after operation (P<0.05).ConclusionFor patients with ACL rupture, ACL reconstruction with the autologous ipsilateral PLT under arthroscopy has satisfactory effectiveness of quick recovery, good function, and great stability.
ObjectiveTo discuss the effectiveness of posterior cruciate ligament (PCL) reconstruction with autologous peroneus longus tendon under arthroscopy.MethodsBetween January 2016 and December 2018, 46 patients with PCL injuries were enrolled. There were 34 males and 12 females, with an average age of 40.7 years (range, 20-58 years). There were 43 cases of acute injury and 3 cases of old injury. The anterior drawer test and the posterior tibia sign were positive in 4 cases, the posterior drawer tests and the posterior tibia sign were positive in 46 cases, the varus stress tests were positive in 10 cases, and the valgus stress tests were positive in 6 cases. The difference of dial-test at 30° knee flexion between affected and healthy sides was (5.20±3.91)°. The tibia posterior displacement under posterior stress position was (12.03±2.38) mm. The Lysholm score of the knee joint was 36.68±7.89, the International Knee Documentation Committee (IKDC) score was 33.58±5.97, and the American Orthopaedic Foot and Ankle Association (AOFAS) score of the ankle joint was 97.60±1.85. PCL was reconstructed with autologous peroneus longus tendon under arthroscopy, and the combined meniscus injury, posterolateral complex injury, and anterior cruciate ligament injury were all treated according to the degree of injury.ResultsAll incisions healed by first intention. Forty patients were followed up 12-26 months, with an average of 16.0 months. At last follow-up, the Lysholm score of the knee joint was 84.85±7.03, and the IKDC score was 87.13±6.27, which were significant different from preoperative ones (t=−13.45, P=0.00; t= −39.12, P=0.00); the AOFAS score of ankle joint was 93.98±2.14, which was not significant different from preoperative one (t=8.09, P=0.90). The tibia posterior displacement under posterior stress position was (2.75±1.76) mm and the difference of dial-test at 30° knee flexion between affected and healthy sides was (1.75±2.09)°, which were significant different from preoperative ones (t=29.00, P=0.00; t=4.96, P=0.00). The posterior drawer test and the posterior tibia sign were positive in 1 case and negative in 39 cases; the anterior drawer test and the varus and valgus stress tests were all negative.ConclusionReconstruction of PCL with autologous peroneus longus tendon under arthroscopy can significantly improve the stability and function of the knee joint, with satisfactory clinical results.
目的 检测类风湿关节炎（RA）患者血清和关节液白细胞介素17A（IL-17A）的变化，探讨其与临床炎症指标、疾病活动性的关系。 方法 2011年6月－2012年6月采用酶联免疫吸附试验检测30例活动性RA患者和20例健康对照血清IL-17A水平，其中18例有膝关节积液RA患者同时检测配对血清和关节液IL-17A水平。 结果 RA组患者血清IL-17A水平显著高于健康对照组[（40.651 ± 16.402）、（23.799 ± 10.693） pg/mL，P＜0.05]。RA患者关节液IL-17A水平明显高于其血清中水平[（63.555 ± 23.405）、（43.727 ± 17.212） pg/mL，P＜0.05]。RA患者血清IL-17A水平只与疾病活动性评分（DAS28）呈正相关（r=0.498，P=0.020），而RA患者关节液IL-17A水平与DAS28和血清C反应蛋白有相关性（r=0.515，P=0.029；r=0.498，P=0.035）。 结论 RA患者血清和关节液IL-17A水平与疾病活动性显著相关，提示IL-17A可作为衡量疾病活动和关节损伤的标志之一。
ObjectiveTo analyze the effectiveness and problems existing in implementing acquired immune deficiency syndrome (AIDS) control and prevention in community health service centers, and to provide scientific evidence for promoting AIDS control and prevention. MethodsRelated information on AIDS control and prevention in community health service centers in a community in the whole year of 2013 was acquired. The effectiveness and problems existing in case management, laboratory testing and highly active antiretroviral therapy (HAART) based on the current community health service system were analyzed. ResultsBy the end of 2013, the rate of AIDS case management was from zero to 100%, the rate of CD4 T detection was from 76.60% (360/470) to 88.35% (508/575), and the rate of receiving highly active antiretroviral therapy among AIDS patients was from 81.40% (175/215) to 84.41% (287/340). But in the actual work on AIDS in community health service centers, there were still some common problems needing to be solved. ConclusionThe community level management mode of AIDS can promote the accuracy of AIDS patients' information, improve the rate of case management, the rate of CD4 T detection and HAART. It is suggested that the community level management mode of AIDS should be promoted in the cities where AIDS patients are concentrated.
ObjectiveTo analyze retrospectively the risk factors of nonarteritic anterior ischemic optic neuropathy (NAION). MethodsThe complete clinical data of 116 patients (134 eyes) were collected. All patients were asked in detail about the disease history and symptoms and were examined for the visual acuity, intraocular pressure, fundus, visual field and fundus fluorescein angiography (FFA), blood pressure, blood glucose, blood fat and head MRI or CT. Suspicious cases and patients with incomplete clinical data were excluded. The relationship between NAION and age, visual field, FFA, systemic and ocular factors, onset seasons were retrospectively analyzed. Results80 patients (68.97%) were 55 to 70 years old. 97 patients (83.7%) had systemic diseases, including 38 patients (39.2) with diabetes mellitus, 32 patients (32.9%) with hypertension (8 patients had low blood pressure at night), 28 patients (28.9%) with hyperlipidemia, 16 patients (16.5%) with cerebrovascular diseases (mainly lacunar cerebral infarction), 6 patients (6.2%) with coronary heart disease. There were 8 patients with ocular factors, including 3 patients (2.6%) with cataract surgery history, 5 patients (4.2%) with small optic discs. The difference of percentage of with or without diabetes mellitus and hypertension was significant (χ2=362, 259; P < 0.05). There were 27.6% patients with disease onset at March to April, 24.1% patients with disease onset at September to October, much higher than other months (χ2=580, P < 0.05). Visual field test results showed that 49 eyes (36.5%) had inferior visual field defect, 12 eyes (9.0%) had superior visual field defect. FFA showed that in the early stage 103 eyes (76.9%) had optic weak fluorescence, 13 eyes (9.7%) had strong fluorescence; in the late stage, 110 eyes (82.1%) had strong fluorescence, 8 eyes (6.0%) had weak fluorescence. ConclusionsDiabetes mellitus, hypertension may be the system risk factors of NAION. The seasonal variation from spring to summer and from autumn to winter may also be another risk factor for the onset of NAION.
ObjectiveTo investigate the efficacy of sulfasalazine combined with thalidomide in the treatment of spondyloarthropathy (SpA), and to probe whether the treatment can reach and maintain clinical remission for the patients. MethodBetween January 2011 and June 2013, we used a prospective, non-intervention and regular follow-up study to observe and assess 70 SpA patients on their Bath ankylosing spondylitis disease activity index, visual analogue scale score, peripheral arthritis, blood sedimentation, and C-reactive protein. All the patients had taken sulfasalazine, thalidomide and non-steroidal anti-inflammatory drugs for 24 weeks. Multivariate analysis of factors affecting the efficacy of the program was our object of this study. ResultsAfter 24 weeks, the total clinical remission rate of these patients was 72.9%. Clinical remission rate of 40 patients with short duration of SpA was 90.0%, while it was 50.0% for the other 30 patients with a non-short duration of SpA. Observation indexes before and after treatment in both groups of patients had significant differences (P<0.05). For patients with non-short duration SpA, the curative effect for female was obviously better than male, but the difference between male and female patients with short-duration SpA was not so obvious. Clinical remission rate for youth was similar with that for non-youth patients. Binary logistic regression analysis showed that whether the disease had a short duration[OR=3.408, 95%CI (1.637, 7.437), P=0.001] and whether the patients were urban residents[OR=4.163, 95%CI (2.011, 8.761), P=0.001] were statistically significant (P<0.05). ConclusionsClinical remission of spondyloarthropathy can be maintain by sulfasalazine combined with thalidomide. Clinical remission rate of the scheme is affected by duration of disease and residency except age and gender of the patients. Short duration and urban residency are independent factors for reaching clinical remission after treatment.