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.
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.
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.
Objective To improve the knowledge of lung injury induced by rituximab. Methods Clinical data of 5 lymphoma patients with lung injury caused by rituximab chemotherapy were analyzed. Results Five patients received chemotherapy including rituximab, and had fever, cough and dyspnea after 3 to 5 chemotherapy cycles. Chest CT showed bilateral diffuse interstitial infiltrates. All 5 cases experienced hypoxemia or respiratory failure. Clinical symptoms were improved 3 to 5 days after the treatment of glucocorticoids, and pulmonary lesions were significantly alleviated 1 to 2 weeks after the treatment. According to the literature, the incidence rate of lung injury caused by rituximab was 0. 03% to 4. 9%, which has increased recently. Conclusions With the comprehensive application of rituximab, lung injury caused by this drug is not rare. The good prognosis depends on early diagnosis and treatment by further recognition of the side effect of rituximab.
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 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 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.
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 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.
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.