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.
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 study the feasibility of constructing tissue engineered cartilage by differentiated rabbit bone marrow mesenchymal stem cells(MSC) cultured in vitro and in vivo. METHODS The MSC were isolated from the nucleated cells fraction of autologous bone marrow by density gradient centrifuge, and then induced into chondrogenic differentiation by adding dexamethasone, transforming growth factor-beta 1 (TGF-beta 1) and ascorbic acid in vitro. After 3 weeks, some cells turned to round shape and secreted metachromatic matrix. The cartilaginoid grafts composed of chondrogenic MSC. Bovine type I collagen and human fibrin were cultured within the chondrogenic medium for 2 weeks in vitro or transplanted subcutaneously adjacent to the knee joint for 3 weeks in vivo. RESULTS The most cells in the grafts were degenerated and disappeared after cultured in vitro. But the residual cells were survival and secreted metachromatic staining proteoglycan with toluidine blue, which was characteristic cartilage matrix. The grafts developed into matured cartilage tissue assessed by histological examination after 3 weeks of transplantation in vivo. CONCLUSION MSC can be used as functional cells to constructing tissue engineered cartilage.
Driven by advances in intelligent technology, artificial intelligence (AI) is emerging as the cornerstone of neurosurgical education. By providing personalized learning experiences and enhancing learning outcomes, AI has enriched the avenues and depth of knowledge acquisition for medical students. The integration of AI not only helps medical students master the basic theories and practical skills of neurosurgery more thoroughly, but also lays a solid foundation for them to provide high-quality and efficient medical services in the future. At the same time, the ability of educators to use intelligent technologies further enhances the interactivity and effectiveness of teaching. In order to further ensure the application of AI in neurosurgery teaching, this article explores the strategic integration of AI in neurosurgical education, emphasizing its critical importance in ensuring that teaching methods evolve with the times.
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.
Acute kidney injury (AKI) is characterized by a sudden and rapid decline of renal function and associated with high morbidity and mortality. AKI can be caused by various factors, and ischemia-reperfusion injury (IRI) is one of the most common causes of AKI. An increasing number of studies found out that exosomes of mesenchymal stem cells (MSCs) could alleviate IRI-AKI by the adjustment of the immune response, the suppression of oxidative stress, the reduction of cell apoptosis, and the promotion of tissue regeneration. This article summarizes the effect and mechanism of MSC-derived exosomes in the treatment of renal ischemia-reperfusion injury, in order to provide useful information for the researches on this field.
目的 回顾性研究老年系统性红斑狼疮（SLE）患者合并感染的危险因素。 方法 选取1995年1月－2009年12月间在四川大学华西医院确诊为SLE，起病年龄为50周岁以上的158例患者，收集性别、临床表现、疾病活动度、实验室检查指标、合并症以及并发症等进行单因素分析或多因素非条件logistic回归分析。 结果 所纳入的158例患者中，合并感染53例（占33.5%），采用单因素分析显示疾病活动性（P=0.001）、低蛋白血症（P=0.030）、糖尿病（P=0.003）、肺间质病变（P=0.000）与老年SLE患者感染发生有关。经logistic回归分析显示，疾病活动性（OR=7.533，P=0.000）、肺间质病变（OR=19.762，P=0.000）、糖尿病（OR=6.862，P=0.025）是老年SLE患者感染发生的危险因素。 结论 积极控制老年SLE的疾病活动度，减少危险因素的发生是控制老年SLE患者并发感染的有效手段。
Superficial temporal artery (STA) - middle cerebral artery (MCA) bypass surgery has been widely used to treat patients with moyamoya disease, and its application value in symptomatic internal carotid artery (ICA)/MCA stenosis/occlusion remains controversial. With the development of imaging, micro-devices and surgical techniques, and the deepen understanding of diseases, the effectiveness of STA-MCA bypass surgery in the treatment of symptomatic ICA/MCA stenosis/occlusion is further required. This article reviews the process of development and evolution of this surgical technique, as well as the significance and deficiencies of several randomized controlled trials of ICA/MCA treatment in the past, and looks forward to possible improvements in future research, so as to clarify the way for further randomized controlled study.
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.
Objective To explore whether bundled care for anesthesia management can reduce the risk of postoperative nausea and vomiting (PONV). Methods The data of laparoscopic cholecystectomy patients admitted to the Day Surgery Center of West China Hospital, Sichuan University between July and November 2021 were retrospectively collected. Patients were divided into a bundled care group and a control group based on whether anesthesia management was implemented according to the bundled care. The demographic characteristics, intraoperative anesthesia management methods, postoperative conditions, and incidence of PONV between the two groups of patients were analyzed and compared. Results A total of 314 patients were included. Among them, there were 124 cases in the bundled care group and 190 cases in the control group; PONV occurred in 52 cases, the incidence of PONV was 16.6% (52/314). Except for surgical time and postoperative incision infiltration (P>0.05), there were statistically significant differences in age, gender, body mass index, anesthesia time, airway establishment, and postoperative analgesic use between the two groups of patients (P<0.05). There was no statistically significant difference in the occurrence of PONV between the bundled care group and the control group (17 vs. 35 cases; χ2=1.205, P>0.05). The results of logistic regression analysis showed that PONV was correlated with gender [odds ratio=0.107, 95% confidence interval (0.030, 0.375), P<0.001], and using bundled care [odds ratio=0.388, 95% confidence interval (0.169, 0.894), P=0.026]. Conclusions Women are at high risk of PONV among patients undergoing day laparoscopic cholecystectomy. The risk of PONV is lower when using bundled care.