【摘要】 目的 检测B细胞成熟抗原（BCMA）mRNA在系统性红斑狼疮（SLE）患者外周血单个核细胞（PBMC）的表达水平，探讨BCMA在SLE发病中的意义。 方法 纳入2006年1-11月收治的36例SLE患者，同期17例健康志愿者作为对照组，采用半定量RT-PCR法检测外周血单个核细胞中BCMA mRNA的表达，并与SLE疾病活动指数（SLEDAI）进行相关性分析。 结果 SLE患者组BCMA mRNA表达水平（0.598±0.230）均明显高于正常对照组（0.411±0.309）（Plt;0.05）。SLE患者BCMA mRNA表达水平与SLEDAI评分无相关性（P=0.590）。 结论 SLE患者BCMA mRNA表达水平的增高，可能在SLE的发病机制中具有一定的作用。【Abstract】 Objective To detect the mRNA expression of B-cell maturation antigen (BCMA) in peripheral blood mononuclear cells (PBMC) in patients with systemic lupus erythematosus (SLE), and explore the role of BCMA in the pathogenesis of SLE. Methods From January 2006 to November 2006 the expression of BCMA mRNA in PBMC of 36 patients with SLE and 17 normal controls were measured by half-quantitative RT-PCR. The linear correlation between the expression of BCMA mRNA and SLE disease activity index (SLEDAI) was assessed. Results The level of BCMA mRNA (0.598±0.230) in PBMC significantly increased in SLE patients compared with that in the normal controls (0.411±0.309) (Plt;0.05). The expression of BCMA mRNA in SLE patients showed no correlation with SLEDAI score (P=0.590). Conclusion The results suggest that the expression of BCMA mRNA might play an important role in the pathogenesis of SLE.
The way of intravenous drug abuse is to puncture the peripheral blood vessels and inject the drug directly into the blood. Therefore, this method has an impact on the peripheral artery and venous system of the users, and can cause a variety of peripheral vascular diseases, such as phlebitis, deep vein thrombosis, chronic venous insufficiency, phlebangioma, atherosclerosis, acute arterial ischemia, pseudoaneurysm, etc. However, due to the particularity of drug abusers, the vascular complications caused by intravenous drug abuse have not attracted enough attention. This paper reviewed the types and pathogenesis of peripheral vascular diseases caused by intravenous drug abuse, so as to improve the clinical understanding of peripheral vascular diseases caused by intravenous drug abuse, improve the prognosis of patients, reduce occupational exposure of medical staff, and play a certain role in social warning.
Objective To investigate the changes of indocyanine green retention rate at 15 minutes (ICGR15) of autologous peripheral blood CD34+ hematopoietic stem cells transplantation in end-stage liver disease (end-stage liver, disease, ESLD) patients with different Child-Pugh grades during before and after transplantation of 3, 6, 12, 36, and 60 months. Methods The CD34+ hematopoietic stem cells transplantation were performed in 60 cases of advanced liver cirrhosis with different Child-Pugh grades who were ineffectively treated with strictly conservative treatment and complied with the criterion of liver transplantation. The ICGR15 were performed before transplantation and in 3, 6, 12, 36 and 60 months after transplantation. And the results of each time point in each Child-Pugh classification group were compared, and the rate of change of ICGR15 value were compared between each Child-Pugh classification group. Results The ICGR15 values of the Child-Pugh grading groups all decreased with time. In Child A group, there were respectively significant differences between the 6 months, 12 months, 36 months, and 60 months groups after transplantation and preoperative and 3 months groups after transplantation (P<0.05), but there was no significant difference between preoperative and 3 months group after transplantation (P>0.05), and there was significant difference between the 12 months and the 60 months group after transplantation (P<0.05). As same as Child A group, there were also significant differences between that time groups in the Child B group (P<0.05), but there were also significant differences between the 3 months group after transplantation and preoperative (P<0.05), and there were respectively significant differences between the 6 months and 12 months, 36 months, and 60 months group after transplantation in the Child B group (P<0.05). Also in the Child C group, there were significant differences between that time groups (P<0.05), but there was no significant difference between preoperative and 3 months group after transplantation (P>0.05), and there were respectively significant differences between the 6 months and 12 months, 36 months, and 60 months group after transplantation (P<0.05). There was no significant difference in the rate of ICGR15 between Child-Pugh classification groups. Conclusion Autologous peripheral blood CD34+ hematopoietic stem cells transplantation can effectively improve the liver function reserve capacity of ESLD patients and improve the safety of operation for a long time.
Objective To investigate the efficiency and safety of autologous peripheral blood stem cell transplantation （ABSCT） in treatment for thromboangiitis obliterans. Methods Fifty patients （62 affected limbs） with thromboangiitis obliterans were treated by ABSCT. A series of subjective indexes including improvement of pain and cold sensation and objective indexes including intermittent claudication distance， ankle brachial index （ABI）， skin temperature， and improvement of foot skin ulcer were evaluated. Results Due to necrosis in middle and lower part of leg， 4 of 50 patients （4 lower limbs） were taken extremity amputation on 3 weeks after ABSCT， 46 patients kept their legs successfully. On 1 month after ABSCT， the legs pain and cold sensation of 46 patients （58 affected limbs） vanished， and the score of feet pain and cold sensation after ABSCT were better than those before ABSCT （P＜0.05）. The intermittent claudication distance， skin temperature， and ABI of 46 patients with kepting their legs on 3 months after ABSCT significantly increased as compared with before ABSCT 〔intermittent claudication distance：（80.38±45.53） m versus （330.56±142.31） m；skin temperature：（26.50±0.46） ℃ versus （31.49±0.45） ℃；ABI：0.41±0.02 versus 0.71±0.05〕， the differences were statistically significant（P＜0.05）. Six months after ABSCT， different degree neonatal lateral vessels were found in 58 affected limbs of 46 patients by lower extremity arteriography. The complications were not found in all the patients by laboratory or CT detection， such as malignant tumors， retinal hyperplasia， aneurysm and so on. After ABSCT， 40 patients were followed up for 9 to 36 months （mean 22.5months）， the symptom had improved. Due to leg pain aggravated after 6 months， score of pain feelings was 4 in 6 patients and with toe ulcers， who had ABSCT again. Eighteen months after transplantation， the patients had only debilitation of lower extremity. The pain feeling was improved （score of pain feeling was 1）. The toe ulcer was healed and no angiosclerotic myasthenia happened. Conclusions ABSCT is a simple， safe， and effective method， especially in treatment for patients with severe lower limb ischemia who is no arterial reconstruction is feasible. It could improve the quality of life of patients and might be avoided amputation of lower extremity or foot.
ObjectiveTo investigate the predictive value of the preoperative peripheral blood neutrophil percentage-to-albumin ratio (NPAR) for survival after radical resection of hepatocellular carcinoma (HCC) and to construct a nomogram prediction model based on NPAR. MethodsAccording to inclusion and exclusion criteria, the HCC patients with China Liver Cancer Staging (CNLC) stage Ⅰa–Ⅱa who underwent radical hepatectomy at West China Hospital of Sichuan University from January 2010 to December 2020 were retrospectively collected and then randomly divided into a training set and a validation set with a 7∶3 ratio. The optimal cutoff value for NPAR was determined using X-tile. Cox proportional hazards regression model was used to identify the independent risk factors for overall survival (OS) in HCC patients and then construct a nomogram model. The predictive performance of the model was evaluated using the C-index, receiver operating characteristic (ROC) curve, and calibration curve, as well as validated in the validation set. ResultsA total of 3 423 HCC patients with CNLC stage Ⅰa–Ⅱa were enrolled in this study, with 2 397 in the training set and 1 026 in the validation set. There were no statistically significant differences in baseline characteristics between the training and validation sets (P>0.05). The optimal cutoff value for NPAR was 17.0, and patients with NPAR ≤17.0 (2 124 cases) had significantly better OS and relapse-free survival (RFS) than those with NPAR>17.0 (273 cases). The multivariate Cox proportional hazards regression model analysis showed that the alpha-fetal protein>400 μg/L, NPAR>17.0, multiple tumors, tumor diameter >5 cm, poor tumor differentiation, capsular invasion, microvascular invasion, and satellite lesions were the independent risk factors affecting postoperative OS in HCC patients (RR>1, P<0.05). The nomogram constructed based on these risk factors demonstrated good discriminations for OS and RFS (C-indexes of 0.708 and 0.709, respectively) and predictive performance in both the training and validation sets. ConclusionsPreoperative high NPAR (>17.0) in HCC patients with CNLC Ⅰa–Ⅱa stages is associated with significantly worse OS compared to those with low NPAR (≤17.0). The nomogram prediction model based on NPAR can effectively predict postoperative survival.
Abstract： Objective To evaluate the sensitivity, specificity and clinical significance of Lunx mRNA in surveying micrometastasis by sampling peripheral blood of lung cancer patients, studying the early diagnosis of lung cancer metastasis. Methods From March 2004 to February 2005，Reverse transcriptionpolymerase chain reaction(RT-PCR) was used to detect Lunx mRNA of peripheral blood of 60 lung cancer patients（lung ancer group）. Peripheral blood of 20 patients with pulmonary benign lesions (pulmonary benign lesions group) and 10 normal healthy volunteers (control group) were used as control. Results (1) In the lung cancer group, Lunx mRNA were expressed positive in 28(46.7％) patients. All the pulmonary benign lesions group (0/20) and the control group (0/10) were expressed negative. (2) One of the 12 stage I patients with lung cancer (8.3％) was positive for Lunx mRNA, 5 of the 15 stage Ⅱ patients (33.3%) were positive, 22 of the 33 stage Ⅲ patients (66.7%) were positive. Comparing the positive rate of these groups, there was no statistically difference between stage Ⅰ and stage Ⅱ, but the difference between stage Ⅰ+ stage Ⅱ and stage Ⅲ significant (χ2=15.88, P=0.000). (3) In 38 adenocarcinoma, 17 were positive for Lunx mRNA. In 14 squamous carcinoma, 7 were positive. All the 3 adenosquamous carcinoma expressed positive. 1 of 3 small cell lung cancer was positive, 1 large cell carcinoma and 1 carcinoma sarcomatodes expressed negative. Comparing the positive rate of these groups, there was no statistically difference among them. (4) By followup till March 2005, 10 lung cancer patients were found metastasis. Among them, 9 were positive for Lunx mRNA expression, and 1 was negative. Conclusion Lunx mRNA has high sensitivity and specificity in surveying micrometastasis by ampling peripheral blood. It would likely to be an proper gene for the detection of micrometastasis in lung cancer patients.
ObjectiveTo separate peripheral blood mesenchymal stem cells (PBMSC) and peripheral blood endothelial progenitor cells (PBEPC) from peripheral blood, and investigate the biological characteristics of composite cell sheets of PBMSC and PBEPC.MethodsThe peripheral blood of healthy adult New Zealand white rabbits was extracted and PBMSC and PBEPC were separated by density gradient centrifugation. Morphological observation and identification of PBMSC and PBEPC were performed. The 3rd generation of PBMSC and PBEPC were used to construct a composite cell sheet at a ratio of 1∶1, and the 3rd generation of PBMSC was used to construct a single cell sheet as control. The distributions of cells in two kinds of cell sheets were observed by HE staining. In addition, the expression of alkaline phosphatase (ALP), osteocalcin (OCN), and vascular endothelial growth factor (VEGF) in the supernatants of cell sheets were observed by ELISA at 1, 5, and 10 days after osteogenic induction.ResultsThe morphology of PBMSC was spindle-shaped or polygonal, and PBMSC had good abilities of osteogenic and adipogenic differentiation. The morphology of PBEPC was paved stone-like, and the tube-forming test of PBEPC was positive. Two kinds of cell sheets were white translucent. The results of HE staining showed that the composite cell sheet had more cell layers and higher cell density than the single cell sheet. The expressions of ALP, OCN, and VEGF in the supernatant of the two groups of cell sheets increased with the time of induction. The expression of OCN in the group of composite cell sheet was significantly higher than that in the group of single cell sheet on the 5th and 10th day, ALP on the 10th day was significantly higher than that in the group of single cell sheet, VEGF expression on the 1st, 5th, and 10th day was significantly higher than that in the group of single cell sheet, all showing significant differences (P<0.05), and there was no significant difference between the two groups at other time points (P>0.05).ConclusionPBMSC have stable differentiation ability, and they have good application prospects because of their minimally invasive access. Composite cell membranes constructed by co-culture of two kinds of cells and induction of membrane formation provides a new idea and exploration for tissue defect repair.
ObjectiveTo research the effect of recombinant adenovirus-bone morphogenetic protein 12 (Ad-BMP-12) transfection on the differentiation of peripheral blood mesenchymal stem cells (MSCs) into tendon/ligament cells. MethodsPeripheral blood MSCs were isolated from New Zealand rabbits (3-4 months old) and cultured in vitro until passage 3. The recombinant adenoviral vector system was prepared using AdEasy system, then transfected into MSCs at passage 3 (transfected group); untransfected MSCs served as control (untransfected group). The morphological characteristics and growth of transfected cells were observed under inverted phase contrast microscope. The transfection efficiency and green fluorescent protein (GFP) expression were detected by flow cytometry (FCM) and fluorescence microscopy. After cultured for 14 days in vitro, the expressions of tendon/ligament-specific markers were determined by immunohistochemistry and real-time fluorescent quantitative PCR. ResultsGFP expression could be observed in peripheral blood MSCs at 8 hours after transfection. At 24 hours after transfection, the cells had clear morphology and grew slowly under inverted phase contrast microscope and almost all expressed GFP at the same field under fluorescence microscopy. FCM analysis showed that the transfection efficiency of the transfected group was 99.57%, while it was 2.46% in the untransfected group. The immunohistochemistry showed that the expression of collagen type Ι gradually increased with culture time in vitro. Real-time fluorescent quantitative PCR results showed that the mRNA expressions of the tendon/ligament-specific genes (Tenomodulin, Tenascin-C, and Decorin) in the transfected group were significantly higher than those in untransfected group (0.061±0.013 vs. 0.004±0.002, t=-7.700, P=0.031; 0.029±0.008 vs. 0.003±0.001, t=-5.741, P=0.020; 0.679±0.067 vs. 0.142±0.024, t=-12.998, P=0.000). ConclusionAd-BMP-12 can significantly promote differentiation of peripheral blood MSCs into tendon/ligament fibroblasts and enhance the expressions of tendon/ligament-specific phenotypic differentiation, which would provide the evidence for peripheral blood MSCs applied for tendon/ligament regeneration.