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.
目的 回顧性研究老年系統性紅斑狼瘡（SLE）患者合并感染的危險因素。 方法 選取1995年1月－2009年12月間在四川大學華西醫院確診為SLE，起病年齡為50周歲以上的158例患者，收集性別、臨床表現、疾病活動度、實驗室檢查指標、合并癥以及并發癥等進行單因素分析或多因素非條件logistic回歸分析。 結果 所納入的158例患者中，合并感染53例（占33.5%），采用單因素分析顯示疾病活動性（P=0.001）、低蛋白血癥（P=0.030）、糖尿病（P=0.003）、肺間質病變（P=0.000）與老年SLE患者感染發生有關。經logistic回歸分析顯示，疾病活動性（OR=7.533，P=0.000）、肺間質病變（OR=19.762，P=0.000）、糖尿病（OR=6.862，P=0.025）是老年SLE患者感染發生的危險因素。 結論 積極控制老年SLE的疾病活動度，減少危險因素的發生是控制老年SLE患者并發感染的有效手段。
Abstract：?Objective?To evaluate clinical outcomes of single utility port complete video-assisted thoracoscopic surgery （VATS） lobectomy for patients with early-stage lung cancer.?Methods?We retrospectively analyzed the clinical data of 162 consecutive patients with early-stage lung cancer who underwent single utility port complete VATS lobectomy from September 2009 to October 2011 in Chinese PLA General Hospital （single utility port group），and compared them with 221 patients with early-stage lung cancer who underwent video-assisted mini-thoracotomy （VAMT） lobectomy in the same period （VAMT group）. The clinical outcomes including operation time, intraoperative blood loss, lymph node dissection number, time to first activity out of bed, chest drainage duration and postoperative complications, were compared between the two groups.?Results?No perioperative death was observed in both groups. There were statistical differences in the intraoperative blood loss （162.8±75.6 ml vs. 231.4±62.8 ml）, time to first activity out of bed （2.2±0.3 d vs. 3.7±0.5 d） , and chest drainage duration （3.5±0.2 d vs. 4.6±0.4 d） between the two groups （P＜0.05）. There was no statistical difference in operation time （133.7±22.0 min vs. 124.9±25.7 min） , lymph node dissection number （11.7±1.9 vs. 12.5±2.7）， and incidence of serious postoperative complications （7.4% vs. 8.1%）between the two groups.?Conclusion?Single utility port complete VATS lobectomy and lymph node dissection are safe and reliable for patients with early-stage lung cancer with less injury and better postoperative recovery compared with VAMT.
Objective To investigate the impacts of cytokines （interleukin-4，IL-4；tumor necrosis factor-α，TNF-α） and medications of bronchial asthma （dexamethasone，aminophylline，salbutamol） on the activity of histamine N-methyltransferase（HMT） in tracheal epithelial cells.Methods BEAS-2B bronchial epithelial cells were cultured and treated with different concentration of TNF-α, IL-4, dexamethasone, salbutamol and aminophylline respectively. The activity of HMT in BEAS-2B cells was determined by high performance liquid chromatography.Results The activity of HMT in tracheal epithelial cells was (50±7) pmol?min-1?mg pro-1.TNF-α and IL-4 lowered the activity of HMT significantly at the concentration equal to or higher than 1 ng/mL and 5 ng/mL respectively，and reached the maximum inhibitory effect at the level of 10 ng/mL.Dexamethasone and aminophylline could ameliorate distinctly the inhibitory effect of TNF-α on the activity of HMT, while salbutamol had no significant inhibitory effect.Conclusions TNF-α and IL-4 exert the lowering effect on the activity of HMT，which would be one important cause of airway hyperreactivity.Glucocorticoids and theophyllines are administered to treat asthma partly due to its relieving mechanism of TNF-α negative effects on HMT.
ObjectiveTo explore the value and role of post-processing techniques such as 3D reconstruction in the online education mode in neurosurgery undergraduate clinical probation teaching.MethodsA retrospective analysis method was used to collect 120 clinical 5-year medical students who were trained in neurosurgery at West China Hospital of Sichuan University from January 2019 to May 2020, including 40 students receiving traditional imaging materials offline (control group 1), 40 students being taught on image post-processing technology offline (control group 2), and 40 students being taught on-line image post-processing technology during the novel coronavirus epidemic (observational group). The students’ scores of departmental rotation examination and feedback survey results on teaching satisfaction were collected, and multiple comparison was conducted between the observational group and the two control groups respectively.ResultIn the control group 1, the control group 2, and the observational group, the theoretical test scores were 36.80±3.22, 38.17±2.61, and 38.97±2.79, respectively; the case analysis scores were 37.05±2.01, 38.40±2.62, and 39.25±2.88, respectively; the total scores were 73.85±5.06, 76.57±4.29, and 78.10±4.53, respectively; the scores of interest in teaching were 84.47±3.71, 86.05±2.87, and 86.82±2.60, respectively; the scores of mastery of knowledge were 82.85±4.39, 84.90±2.72, and 85.78±2.36, respectively; and the scores of overall satisfaction with teaching were 84.17±3.45, 85.97±2.64, and 86.37±2.59, respectively. The differences among the three groups were all statistically significant (P<0.05). The observational group differed significantly from the control group 1 in all the above scores (P<0.05), while did not differed from the control group 2 in any of the above scores (P>0.05).ConclusionsIn neurosurgery internship activities, the online application of image post-processing techniques such as 3D reconstruction will help students establish 3D spatial concepts, better understand the brain anatomy, and improve students’ academic performance and acceptance.
Objective To identify the the key factors influencing the growth of national-level high-caliber talents in medical disciplines through multi-angle and comprehensive research. Methods A questionnaire survey was conducted among medical talents from eastern and western regions, and in-depth interviews were carried out with 8 of these individuals from April to June 2024. The questionnaire results were analyzed for reliability and validity, and a comprehensive analysis was performed in conjunction with the interview results. Results A total of 167 medical talents were surveyed. The reliability analysis of the questionnaire showed a Cronbach’s α value of 0.814, indicating high internal consistency. The KMO and Bartlett’s sphericity test results revealed a KMO value of 0.75, with Bartlett’s test of sphericity yielding a P-value<0.001, suggesting suitability for factor analysis. The validity analysis extracted three common factors with eigenvalues greater than 1, accounting for a cumulative variance contribution rate of 67.136%. The three common factors were named as “incentive mechanisms” “global perspective” and “fostering a positive academic atmosphere and cultural environment”, respectively. The findings from the in-depth interviews were largely consistent with the questionnaire survey results. Conclusion This study identifies three main factors influencing the growth of national-level talents in medical disciplines: establishing effective incentive mechanisms, cultivating strategic thinking and a global perspective among national-level talents, and fostering a positive academic atmosphere and cultural environment.
Objective To review research advances of revision surgery after primary total hip arthroplasty (THA) for patients with Crowe type Ⅳ developmental dysplasia of the hip (DDH). Methods The recent literature on revision surgery after primary THA in patients with Crowe type Ⅳ DDH was reviewed. The reasons for revision surgery were analyzed and the difficulties of revision surgery, the management methods, and the related prosthesis choices were summarized. Results Patients with Crowe type Ⅳ DDH have small anteroposterior diameter of the acetabulum, large variation in acetabular and femoral anteversion angles, severe soft tissue contractures, which make both THA and revision surgery more difficult. There are many reasons for patients undergoing revision surgery after primary THA, mainly due to aseptic loosening of the prosthesis. Therefore, it is necessary to restore anatomical structures in primary THA, as much as possible and reduce the generation of wear particles to avoid postoperative loosening of the prosthesis. Due to the anatomical characteristics of Crowe type Ⅳ DDH, the patients have acetabular and femoral bone defects, and the repair and reconstruction of bone defects become the key to revision surgery. The acetabular side is usually reconstructed with the appropriate acetabular cup or combined metal block, Cage, or custom component depending on the extent of the bone defect, while the femoral side is preferred to the S-ROM prosthesis. In addition, the prosthetic interface should be ceramic-ceramic or ceramic-highly cross-linked polyethylene wherever possible. Conclusion The reasons leading to revision surgery after primary THA in patients with Crowe type Ⅳ DDH and the surgical difficulties have been clarified, and a large number of clinical studies have proposed corresponding revision modalities based on which good early- and mid-term outcomes have been obtained, but further follow-up is needed to clarify the long-term outcomes. With technological advances and the development of new materials, personalized prostheses for these patients are expected to become a reality.
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：機械分離、培養小鼠耳蝸螺旋神經元，并進行免疫熒光細胞學鑒定，為后期進一步的實驗研究提供實驗材料。方法：采用初出生1～5天以內的昆明小鼠進行解剖、機械分離以獲得螺旋神經節組織，進行原代培養后，應用神經微絲蛋白（Neurofilament protein,NFP－H）單克隆抗體進行免疫熒光細胞學鑒定。結果：機械分離后獲得的螺旋神經節組織中的螺旋神經元，在體外培養條件下可以存活并進行正常分化。典型的螺旋神經元，其細胞形態呈橢圓形,胞體透明光滑、接近生理形態。熒光染色標記后，胞體和神經突起均顯色好，Schwann細胞和成纖維細胞未著色。結論：應用機械分離的方法獲得小鼠耳蝸螺旋神經節組織并進行培養，耳蝸螺旋神經元在體外可以穩定地存活生長。培養獲得的細胞形態和生存狀態接近生理狀態，滿足電生理、免疫細胞化學、藥理學等研究。應用特異性的神經微絲蛋白對培養獲得的螺旋神經元進行免疫熒光細胞學鑒定，特異性好，熒光顯色好。
Objective To study the influence of different mechanical environments on repair cartilage defect with marrow mesenchymal stem cells as seed cells. Methods The rabbit marrow mesenchymal stem cells were isolated and cultured. The cartilage defects were repaired by autologous tissue engineered cartilage with the marrow mesenchymal stem cells as seed cells. Fifteen rabbits with cartilage defect were divided into 3 groups: dislocation group with cell-free scaffold(controlgroup), dislocation group with cartilaginous construct and normal mechanical environment group with cartilaginous construct. The repaired tissue was harvested and examined 6 weeks postoperatively. Results The repair tissue in normal mechanical environment group with cartilaginous construct showed cartilage-like tissue in superficial layer and subchondral bone tissue in deep layer 6 weeks postoperatively. The defect was filled with bone tissue in dislocation group with cartilaginous construct 6 weeks postoperatively. The surrounding normal cartilage tissue showed vascular invasion from subchondral area and the concomitant thinningof the normal cartilage layer. The cartilaginous construct left in the femoral trochlea groove formed hyaline cartilage-like tissue. The defect was repaired byfibrous tissue in control group. Conclusion The repaired tissue by tissue engineered cartilage with marrow mesenchymal stem cells as seed cells showed the best result in normal mechanical environment group, which indicates that it will be essential for the formation and maintenance of tissue engineered cartilage to keep the normal mechanical stress stimulus.