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.
ObjectiveTo study the contents of CD44 that shared exon variant 5 (CD44v5) in peripheral blood lymphocytes (PBL) of patients with gastric carcinoma and the expression of CD44v5 in tumor tissue and their clinical significance. MethodsThe contents of CD44v5 were determined by FlowCytometry in PBL of 31 patients with gastric carcinoma before surgery and 10 normal controls. Tissue expression of CD44v5 in 33 patients with gastric carcinoma was investigated by immunohistochemistry. ResultsThe contents of CD44v5 were significantly higher in PBL of patients with gastric carcinoma before surgery than those of controls (P＜0.01). Nodepositive gastric cancer patients showed significantly elevated contents of CD44v5 in PBL in comparison with nodenegative gastric cancer (P＜0.01）. Significant correlations were noted between the contents of CD44v5 in PBL of patients with gastric carcinoma before surgery and tumor size, depth of invasion, lymph node metastasis and different the Vnion International Centre Le Cancer (VICC) stages of tumor (P＜0.05). The expression of CD44v5 could be detected in 69.7% of tumor tissue,but was not detected in adjacent normal gastric mucosa. Significant correlations were noted between CD44v5 expression and depth of invasion,and lymph node metastasis.The presence of CD44v5 protein was correlated with the lymph node involvement rate. Conclusion CD44v5 in PBL or tumor tissue may be useful as a metastatic marker. It may be of important clinical value in the diagnosis of metastasis and judgement of development for the patients with gastric cancer.
From March 1979 to February 1987, 500 cases of firearm wounds of blood vessels were treated. Of them, 465 cases were recovered, 15 cases were disabled, 13 cases had amputation, and 7 cases died. The article presented the clinic materials. The following problems were discussed: (1) The characteristics of firearm wounds of blood vessels. (2) Emergency treatment of injuries of major blood vessels of limbs. (3) Indications of repair of blood vessels. (4) The methods of repair of defect in blood vessel. (5)Factors influencing the survival of extremities, and (6) Active prevention and treatment of complication.
Effect of radical operation on expression of interleukin-2(IL-2)mRNA and production of IL-2 were markedly reduced preoperatively and four weeks after operation,expression of IL-2 mRNA significantly enhanced,but it was still lower than that in the normal group.Production of IL-2 nearly reached normal level,When PBL was activated by phytohemagglutinin(PHA),expresseion of IL-2 mRNA and production of IL-2 were much higher than that in non-activated PBL.These results suggested that expression of IL-2 were much higher than that in non-activated PBL.These results suggested that expression of IL-2 mRNA and production of IL-2 are dificient in gastric cancer patients,and radical surgery will help them to recover and they can also be improved through activation with PHA.
ObjectTo investigate the pathogenesis of drug-resistant epilepsy by examining the expression of mRNA and protein of Cell Division Cycle 42 GTP-binding protein (Cdc42), Neural Wiskott-Aldrich Syndrome Protein (N-WASP) and Actin-related protein 2/3(Arp2/3) in peripheral blood of patients with drug-resistant epilepsy (DRE).MethodsSeventy two essential epilepsy patients who were attended at outpatients and inpatients in the Department of Neurology of the Affiliated Hospital of Youjiang Medical University for Nationalities were selected from October 2016 to October 2018. According to the 2010 International League Against Epilepsy’s definition of Drug-Resistant Epilepsy, the patients were divided into 2 groups: 32 patients with DRE were defined as DRE group, 40 patients with anti-epilepsy drugs (AEDs) well controlled were defined as the well controlled group. Thirty two healthy persons were selected as control group. The expression of mRNA and protein of Cdc42, N-WASP and Arp2/3 in peripheral blood were measured by quantitative real-time PCR (RT-qPCR) and Western blot(WB). Experimental data were analyzed by ANOVA or rank-sum test.ResultsCompared with well-controlled group and healthy persons group, Cdc42, N-WASP, Arp2/3 in DRE group were significantly increased, the differences were statistically significant (P<0.05). Compared with the control group, Cdc42, N-WASP, Arp2/3 in well-controlled group were significantly increased, with statistically significant differences (P<0.05).ConclusionThe expression of Cdc42, N-WASP, Arp2/3 in peripheral blood of patients with DRE significantly increased, being closely related to the occurrence and development of DRE, and used as indicators in peripheral blood predicting the occurrence of DRE.
ObjectiveTo investigate the clinical significancy of K-ras gene mutation in peripheral blood free DNA in patients with non-small cell lung cancer (NSCLC). MethodsA total of 242 patients pathologically diagnosed with NSCLC in the Third People's Hospital of Chengdu were recruited between January 2013 and August 2015. Both tumor tissues and peripheral blood free DNA were collected for detection of K-ras gene mutation by mutant-enriched liquidchip technology. The detection rate was compared between these two kinds of samples. ResultsIn tumor tissues, the K-ras gene mutation was detected in 12 cases with a positive rate of 4.96%. While in peripheral blood samples, the K-ras gene mutation was detected in 10 cases with a positive rate of 4.13%. The detection yield of K-ras gene mutation in peripheral blood had a good consistency with that of lung cancer tissues (Kappa value=0.81). ConclusionK-ras in peripheral blood plasma free DNA can be a surrogate marker for tumor tissues' K-ras gene mutation in screening patients with NSCLC.
In recent years, many scholars have explored the clinical application value of a number of peripheral hematology indexes in tumor patients. The significant correlation of neutrophil to lymphocyte ratio and platelet to lymphocyte ratio with the prognosis in various tumors has also been confirmed. At present, more peripheral blood indexes have been gradually applied to the evaluation of the prognosis in patients with malignant tumors. Small cell lung cancer (SCLC) is a type of highly malignant tumor and most patients are in advanced stage at the time of diagnosis. The evaluation value of tumor stage for survival is extremely limited. Therefore, this review intends to explain the relationship between various peripheral hematology indexes and the prognosis of SCLC patients, so as to provide some academic evidence for the clinical assessment of the survival of SCLC patients and formulation of appropriate treatment strategy, which may contribute to the improvement of the prognosis.
Objective To compare the effectiveness of autologous implantation between bone marrow stem cells and peripheral blood stem cells for treatment of lower limb ischemia. Methods From December 2004 to December 2005, 42 patients with unilateral lower limb ischemia were treated with both autologous bone marrow stem cell implantation(group A, n=21)and autologous peripheral blood stem cell implantation (group B, n=21). Fortytwo patients included 32 males and 10 females. The age ranged from 34 to 80 years, with a mean of 65.6 years. Of the patients, there were 28, 8 and 6 patients suffered from diabetic lower limb ischemia, Burger’s disese and atherosclotic occlusion, respectively. Ischemic history was from 3 months to 5 years, with amean of 2.1 years. A series of subjective indexes (such as improvement of pain, cold sensation and numbness) and objective indexes such as increase of ankle brachial index (ABI), transcutaneous oxygen pressure (TcPO2), angiography, amputation rate, and improvement of foot wound healing, were used to evaluate the effect. Results After 4 weeks of implantation, the rate of pain relief was 88.2% in group A and 89.5% in group B (Pgt;0.05) ; the rate of cold sensation relief was 94.4% in group A and 94.7% in group B（Pgt;0.05）; improvement of numbness was 69.2% and 66.7% respectively in groups A and B（Pgt;0.05）. Increaseof ABI was 38.1% in group A and 33.3% in group B（Pgt;0.05）; increase of TcPO2 was 85.7% and 90.5% respectively in groups A and B（Pgt;0.05）; angiography was performed in 12 patients (group A) and 9patients (group B), and the new formed collateral vessel rate was 83.3% in group A and 77.8% in group B（Pgt;0.05）; the amputation rate was 9.1% in groups A and B（Pgt;0.05）; the rate of improvement of foot wound healing was 60.0% in group A and 66.7% in group B(P＞0.05). Forty patients were followed up 3-15 months(mean 8 months). The improvement rate of subjective symptoms was 75.0% in group A and 70.0% in group B (Pgt;0.05); increase of ABI was 60.0% in group A and 65.0% in group B; increase of TcPO2 was 80.0% and 75.0% respectively in groups A and B; the new formed collateral vessel rate was 90.0% in group A and 84.6% in group B. All ulcers healed except 1 case in group B. Conclusion Bone marrow stem cell graft and peripheral blood stem cell graft are all effective in treatingower limb ischemia.
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.
Objective To determine the application values of gene chip technique in cardiovascular surgical clinical and research work. Microarray for gene expression profiles was used to screen out the differentially expressed genes during cardiopulmonary bypass(CPB) in peripheral blood mononuclear cell. By doing these, it was hoped that some clues in inflammatory response during CPB could be found out. Methods The patients’ oxygenated bloods were drawn immediately before onset and termination of CPB. Peripheral blood mononuclear cell (PBMC) were obtained from heparinised blood by Ficoll gradient centrifugation. The differentially expression was measured using BD AtlasTM cDNA Expression Arrays. The candidate genes were corroborated by semiquantitative reverse transcriptionpolymerase chain reaction (RT-PCR). Results Gene chip technique was successfully used in CPB study. The gene expression profiles of cytokines of PBMC during CPB were screened out. Interleukin 6 and Wnt5a were the differentially expressed genes. But the validity using semiquantitative RT-PCR found no statistically difference(P=0.888,0.135). Conclusion Microarray technique has positive application values in the study of cytokines during CPB. cDNA microarray for gene expression profiles can primarily screen out differentially expression genes during CPB. These genes may be engaged in inflammation and other pathophysiological reactions during CPB. PBMC is not the major source of cytokines during CPB.