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.
Objective To evaluate the effects of intraocular lenses (IOL) of silicone and polymethylmethacrylate (PMMA) on posterior capsule opacification (PCO). Methods We searched MEDLINE (1966 to 2003), EMBASE (1980 to 2003), Cochrane Central Register of Controlled Trials (Issue 1, 2003) and CBMdisc (1979 to 2003). Only randomized controlled trials (RCTs) were identified. Two reviewers independently assessed the quality of the included trial and extracted data. The following outcomes were assessed: PCO index, percentage of neodymium: YAG (Nd: YAG) laser posterior capsulotomy and visual acuity after cataract surgery. RevMan 4.2 was used for meta-analysis.Results Nine RCTs including 672 eyes were included. In patients with senile cataract, the results of meta-analysis suggested that silicone lens were better than PMMA lens on PCO index (the pooled SMD=－0.92, 95%CI －1.19 to －0.64) and percentage of Nd: YAG (the pooled OR 0.35, 95%CI 0.22 to 0.57) with a statistically significant difference, but there was no significant difference on visual acuity (the pooled OR 1.22, 95%CI 0.43 to 3.50) after cataract surgery between silicone lens and PMMA lens. However, in patients with uveitic cataract, the results of meta-analysis showed that PMMA lens were better than silicone lens in visual acuity (the pooled OR 0.38, 95%CI 0.15 to 0.91) after cataract surgery with a statistically significant difference, but there was no significant difference on PCO index (SMD 0.23, 95%CI －0.59 to 1.05) and percentage of Nd: YAG (the pooled OR 1.82, 95%CI 0.47 to 6.95) between silicone lens and PMMA lens. Conclusions Current evidence indicates that silicone lens are superior to PMMA lens in reducing PCO. Further trials with high quality on methodology are required.
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.
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。
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.
目的：机械分离、培养小鼠耳蜗螺旋神经元，并进行免疫荧光细胞学鉴定，为后期进一步的实验研究提供实验材料。方法：采用初出生1～5天以内的昆明小鼠进行解剖、机械分离以获得螺旋神经节组织，进行原代培养后，应用神经微丝蛋白（Neurofilament protein,NFP－H）单克隆抗体进行免疫荧光细胞学鉴定。结果：机械分离后获得的螺旋神经节组织中的螺旋神经元，在体外培养条件下可以存活并进行正常分化。典型的螺旋神经元，其细胞形态呈椭圆形,胞体透明光滑、接近生理形态。荧光染色标记后，胞体和神经突起均显色好，Schwann细胞和成纤维细胞未着色。结论：应用机械分离的方法获得小鼠耳蜗螺旋神经节组织并进行培养，耳蜗螺旋神经元在体外可以稳定地存活生长。培养获得的细胞形态和生存状态接近生理状态，满足电生理、免疫细胞化学、药理学等研究。应用特异性的神经微丝蛋白对培养获得的螺旋神经元进行免疫荧光细胞学鉴定，特异性好，荧光显色好。
ObjectiveTo explore the effect of fingolimod (FTY720) on secondary nerve injury after thalamic-ventricle hemorrhage (TH-IVH) in rats.MethodsAdult male Sprague Dawley rats (clean animal) were randomly divided into 3 groups: sham group, TH-IVH group, and intervention group (FTY720 group), with 6 rats in each group. TH-IVH model was established in both TH-IVH group and FTY720 group, but only the rats in FTY720 group were treated with FTY720. The observation was conducted at the 1st, 3rd and 7th day after modeling. The main observation index included scores of neurological function, change of body weight, water content of brain tissue, the activation of inflammatory cells, the degree of neuronal degeneration and apoptosis, and the level of cell autophagy.ResultsAt the 1st, 3rd and 7th day after modeling, the change of body weight, the neurological score, brain edema and microglia activation in TH-IVH group were statistically different from those in sham group and FTY720 group (P<0.05). The number of degenerated neurons and the number of apoptotic cells in TH-IVH group were statistically different from those in sham group and FTY720 group at the 1st and 3rd day after modeling (P<0.05). The differences in the ratio of LC3Ⅱ/LC3Ⅰ protein expression andBcl-2/Bax expression were statistically significant between FTY720 group and TH-IVH group at the 1st and 3rd day after modeling (P<0.05).ConclusionsFTY720 can improve neurological function of the TH-IVH model in the acute phase, and has certain neuroprotective effect. The neuroprotective effect of FTY720 may be associated with neuronal autophagy and apoptosis regulation and immunosuppression.
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 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.
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.