Progress of Research on Tumor Extracellular Matrix and Discoidin Domain Receptor 1 / 肿瘤防治研究

Tumor extracellular matrix (ECM) is the center component of tumor microenvironment (TME), ECM diversity constitutes the inherent heterogeneity of TME that contributes to tumor growth, dormancy, drug resistance, and metastasis. Discoidin domain receptor 1 is one of the ECM receptors that interact with multiple ECM ligands. It also regulates the occurrence and development of tumors. Accordingly, DDR1 plays an increasingly important role in the prevention, diagnosis, and treatment of cancer. In this review, we primarily summarize the research of ECM and its receptors with components, regulation, cell receptors, and signaling pathways in tumor progression.

Differential diagnosis and treatment of chylothorax and pseudochylothorax after lung cancer surgery / 中华胸心血管外科杂志

Objective:To explore the differential diagnosis and different treatment methods of chylothorax and pseudochylothorax after lung cancer surgery.Methods:Clinical data of 1 584 surgical patients with non-small cell lung cancer from January 2016 to December 2021 were analyzed, 21 cases of chylothorax and 8 cases of pseudochylothorax were identified and analyzed to compare the differences in pleural fluid chyle test, pleural effusion biochemical values, total cholesterol, triglycerides, total cholesterol/triglyceride ratio, leukocyte count, bacterial culture and treatment.Results:The incidence of chylothorax after lung cancer surgery was 1.3%, and the incidence of pseudochylothorax was 0.5%; 80.9%% of chylothorax on the right side was significantly higher than 19.1% of chylothorax on the left side, and the difference was statistically significant( P<0.05). Pseudochylothorax occurred on the right side(100%). The difference between chylothorax and pseudochylothorax in pleural fluid tests for cholesterol and triglyceride was statistically significant( P<0.05), the leukocyte count was significantly higher in pseudochylothorax than chylothorax, and the difference was statistically significant( P<0.05). The differences in drainage before treatment, postoperative drainage time and postoperative hospitalization time between the two groups were statistically significant( P<0.05). The success rate was 61.9% in 13 cases of chylothorax treated conservatively and 38.1% in 8 cases of thoracic duct clamping; all cases of pseudochylothorax were treated conservatively with a success rate of 100%. Conclusion:In naddition to pleural fluid chyle test and pleural effusion biochemical values, total cholesterol, triglyceride and total cholesterol to triglyceride ratio in pleural fluid should be tested to identify chylothorax and pseudochylothorax, high triglyceride in pleural fluid diagnosed as chylothorax; Pseudochylothorax is diagnosed with a cholesterol/triglyceride ratio >1 in the pleural fluid, pseudochylothorax is usually treated conservatively. Chylothorax is treated conservatively and surgically according to different conditions. If the drainage flow is greater than 800 ml/day for 3 consecutive days or if it causes serious electrolyte disorders, it is recommended to perform thoracoscopic-assisted thoracic duct clamping via right-sided approach.

Constructionof the militaryandlocal integration course of"Field and Disaster Nursing"based on Delphi method / 中国实用护理杂志

Objective To construct a"Field and Disaster Nursing"course which is suitable for the military and local higher nursing education, on-the-job and post training education to organic integrate the modern disaster nursing teaching and military field nursing teaching. Methods Based on the methods of qualitative and quantitative research, such as literature research, interviews and observational method, a preliminary plan of curriculum design was established, and an expert consultation table was designed.The Delphi expert consultation method was used to conduct two rounds of consultation with 30 related experts on disaster relief.Quantitative data were obtained and statistical data were analyzed. Results The final screening results of the curriculum content applicability index are 4 first grade, and 23 for the two level.A set of"Field and Disaster Nursing"curriculum had been set up that is suitable for education and training needs of the military and local fusion. Conclusion The construction of this course provides a training thought for exploring disaster nursing education suitable for China's national conditions, and has reference and guiding significance for further implementation of military and civilian integration and multi participatory disaster nursing teaching practice.

Effect of sorafenib on growth and circadian clock gene expression of human hepatocellular carcinoma in nude mice / 中华肝胆外科杂志

Objective To investigate the function of sorafenib on the growth of hepatocellular carcinoma by establishing subcutaneous transplantation tumor model with nude mice.To explore the effect of sorafenib on circadian clock gene expression in hepatoma cells.Methods Mouse tumor model was established by implanting hepatocarcinoma cell (HepG2) subcutaneously in Balb/C nude mice.Sixteen experimental mice were randomly divided into two groups:sorafenib treatment group (n =8) and solvent control group (n =8).The nude mice were treated with sorafenib (100 mg/kg) and DMSO daily by intragastric administration,respectively.The volume of tumors was recorded every 3 days.The expressions level of circadian clock genes (Per1,Per2,Per3,CLOCK,Cry1,Cry2,BMAL1 and CKIε) were detected by real-time polymerase chain reaction (Real-time PCR).The correlations between the size of the xenografts and the expressions of the circadian clock genes were further analyzed.Results Compared with the control group,the tumor size in the sorafenib treatment group were significantly smaller comparing with the control group.Results of Real-time PCR showed that the expression level of Per1,Cry1 and BMAL1 mRNA was remarkably decreased in the treatment group (Per1,P =0.02;Cry1,P =0.002;BMAL1,P =0.035),the differences were statistically significant.Correlation analysis showed that the size of subcutaneous transplantation tumorsin nude mice was negatively correlated with the expressions of Per1,Per2,Cry1 and Cry2 mRNA in control group.While,the size of subcutaneous transplantation tumors was negatively correlated with the expressions of Per2,Per3 and BMAL1 levels in the sorafenib treatment group.Conclusions There is a negative correlation between the expression levels of some biological clock genes and the size of transplantation tumor in nude mice.Sorafenib treatment significantly inhibited the growth of hepatocellular carcinoma in nude mice and down-regulation the expressions of Per1,Cry1 and BMAL1 mRNA in hepatoma cells.

Research progress of preventive effects of statins on contrast induced nephropathy / 心血管康复医学杂志

Contrast induced nephropathy (CIN)is one of the main complications caused by intravenous iodinated contrast media injection,which can increase hospitalization rate and mortality of patients,and it has become the third main cause of acquired acute renal failure in hospital currently, but its fundamental pathogenesis is still not clear,which is considered to be related to inflammation and oxidative stress response etc.The present article made a review on pathogenesis of CIN and preventive effects of statins on CIN.

Heme oxygenase-1 and research progress in tumor / 实用肿瘤学杂志

Heme oxygenase-1(HO-1)is a type of stress related gene,which harbors the ability of anti-inflammatory,anti-apoptosis,anti-oxidation and drug resistance.In physiological status,the expression of HO-1 gene in human normal tissues is extremely low.When subjected to a constant stimulation of alcohol,spicy and hot,over expression of HO-1 gene in the related tissues is often observed.In tumor tissues,the abnormal expres-sion of HO-1 is closely related to their biological behaviors.In recent years,there is a trend to believe that the gene polymorphism and its metabolic products( bilirubin,Fe2+,CO) involve in the decomposition of heme partici-pate in the activation of many signaling pathways,and affect the initiation and development of tumor diseases.In addition,the expression level of HO-1 gene is up-regulated in many tumor diseases,which can not only affect the growth and proliferation of tumor cells by regulation of the oxidative stress levels of tumor microenvironments, but also induce the tumor cells to produce drug resistance.Therefore,the abnormal expression of HO-1 may be involved in the initiation and development processes of tumors.

Significance of FGFR4 expression in prognosis in lung adenocarcinoma / 实用医学杂志

Objective To investigate the expression of FGFR4 in lung adenocarcinoma tissue and its association with the prognosis of lung adenocarcinoma. Methods The expression FGFR4 in 128 lung adenocarcinoma tissue and the normal lung tissue was detected by the immunohistochemistry SP method.The relationship between the expression of FGFR4 in lung adenocarcinoma and the prognosis of lung adenocarcinoma was analyzed. Results The expression of FGFR4 protein in lung adenocarcinoma was significantly lower than that in the tumor-adjacent normal lung tissue (P 0.05), but was significantly and positively correlated with the degree of tumor differentiation and T stage, N stage and the clinical TNM stage of lung adenocarcinoma (P < 0.05).Kaplan-Meier survival test showed that the postoperative survival time in the high expression group of FGFR4 was significantly longer than that in the low FGFR4 expression group (P < 0.05). Conclusion The expression of FGFR4 protein in lung adenocarcinoma was significantly lower than that in the tumor-adjacent normal lung tissue. Postoperative survival time in the high FGFR4expression group was significantly longer than that in the low FGFR4 expression group.

Clinical Value of CYP2C19 Polymorphisms for Guiding the Anti-platelet Therapy in CAD patients After Percutaneous Coronary Intervention / 中国循环杂志

Objective: To explore the value of CYP2C19 genotype polymorphisms for guiding the strategy of anti-platelet therapy in coronary artery disease (CAD) patients after percutaneous coronary intervention (PCI) with clinical prognosis. Methods: A total of 326 CAD patients with PCI treatment received CYP2C19 genotype examination. According to CYP2C19 polymorphisms, the patients were divided into 3 groups. The patients with CYP2C19*1/*1, fast metabolism were defined as Routine group, who received clopidogrel 75 mg/day,n=128. The patients with CYP2C19*1/*2, CYP2C19*1/*3, CYP2C19*2/*3, CYP2C19*2/*2, CYP2C19*3/*3, medium and slow metabolism were further divided into 2 groups as High clopidogrel group, who received clopidogrel 150 mg/day,n=99 and Ticagrelor group, who received ticagrelor 90 mg/bid,n=99. The changes of platelet aggregation rate (PAgT) at before and 1, 3, 6 months after PCI were observed, the major adverse cardiac events (MACE) and bleeding condition were compared among different groups at 6 months after PCI. Results: The PAgT was similar between High clopidogrel group and Routine group at 1, 3, 6 months after PCI, P>0.05;while compared with High clopidogrel group and Routine group, the PAgT decreased in Ticagrelor group at 1, 3, 6 months after PCI,P0.05; while compared with High clopidogrel group and Routine group, Ticagrelor group had much less MACE occurrence,P0.05. Conclusion: High dose clopidogrel does not increase MACE occurrence in CAD patients with CYP2C19 medium and slow metabolism after PCI. Ticagrelor may decrease PAgT and MACE occurrence without elevating the major bleeding events.

The role of hepatitis B virus X protein in regulation of hypoxia inducible factor-1αand the underlying mechanisms in hepatocellular carcinoma / 中国癌症杂志

Background and purpose:Hepatitis B virus X protein (HBx) and hypoxia inducible factor-1α(HIF-1α) play key roles in hepatocarcinogenesis and the development of hepatocellular carcinoma. Positive correlation on the expression of these 2 proteins in hepatocellular carcinoma tissues has been found, whereas the underlying mechanisms have not been fully elucidated. This study focused on the role of HBx in regulating HIF-1α and the underlying mechanisms in hepatocellular carcinoma cells. Methods:The expression plasmids were transfected into Huh7 cells with LipofectemineTM 2000. Western blot analysis was applied to detect the expressions of HIF-1αand HIF-1β protein. The transcriptional activity of HIF-1α was detected by the commercial analysis kits. The mRNA levels of HIF-1αand its target genes, including vascular endothelial growth factor (VEGF) and multi-drug resistance gene 1 (MDR1), were detected by quantitative real-time PCR (qRT-PCR). Immunoprecipitation analysis was applied to detect the interaction of HIF-1α, HBx and protein von Hippel-Lindau (pVHL). Results:Huh7 cells transfected with HBx plasmid led to sharp increase of HIF-1αprotein and transcriptional activity, as well as the mRNA of VEGF and MDR1 (P0.05). Meanwhile, HBx also signiifcantly impaired the function of pVHL in mediating the degradation of HIF-1αby ubiquitin hydrolase. This finding was further confirmed by the immunoprecipitation analysis, which showed that HBx could directly bind to pVHL, but not to HIF-1α. Conclusion:HBx may inhibit the inter-activation between pVHL and HIF-1αthrough directly binding to pVHL, and thus enhance the stability and transcriptional activity of HIF-1α.

Single-port video-assisted thoracoscopic surgery for pulmonary diseases: analysis of 158 cases / 南方医科大学学报

<p><b>OBJECTIVE</b>To assess the clinical value of single-port video-assisted thoracoscopic surgery (VATS) for treatment of pulmonary diseases.</p><p><b>METHODS</b>From October, 2009 to December, 2013, 105 patients with pulmonary diseases were scheduled for single-pore VATS for pulmonary lobectomy (19 patients), wedge resection of the lung (34 patients), and bullae resection and pleurodesis for spontaneous pneumothorax or pulmonary bleb (52 patients).</p><p><b>RESULTS</b>Of the 105 patients, 101 patients underwent single-port VATS; the procedure was converted to open thoracotomy in 1 patient and to conventional three-port VATS in 2 patients. The operative time was 50.6∓36.8 min (20-200 min) with intraoperative blood loss of 70∓56.9 ml (10-300 ml), thoracic drainage time of 4.2∓3.2 days (2-14 days), and postoperative hospital stay of 5.4∓3.8 days (3-16 days). Postoperative complications of the procedures included prolonged air leakage (6 cases) and atelectasis (2 cases). All the other patients recovered smoothly without serious complications.</p><p><b>CONCLUSION</b>Single-port VATS is a safe and efficient procedure that allows rapid postoperative recovery and is a method of choice for selected patients with pulmonary diseases.</p>

Comparison of radiation doses between rotational angiography and standard angiography / 中华放射医学与防护杂志

Objective To compare radiation doses to patients between rotational angiography and standard angiography.Methods According to different modes,our experiment was divided into RA (Dual-axis Rotational Angiography) group and SA (Standard Angiography) group.The latter was divided into 3 subgroups (SA1,SA2 and SA3) in terms of different time of cine.The procedure time and the position were controlled by the order preset into the machine.Each group consisted of 10 repeated operations with a unique phantom to simulate the true patients.The radiation doses were measured by using dose meter provided by the machine and TLD system; the means of the data were compared.Results The mean of total accumulated DAP and AK in group RA were 3 061.6 mGy·cm2 and 64.2 mGy,respectively,compared with group SA1 (4 213.9 mGy·cm2 and 85.5 mGy),SA2 (6 436.0 mGy·cm2 and 112.2 mGy),SA3 (12 810.4 mGy·cm2 and 243.7 mGy).The mean of accumulated dose in group RA was 37.70 mGy,higher than group SA1 (26.56 mGy) but lower than group SA2 (46.86 mGy) and SA3 (77.79 mGy).The gap between highest dose and lowest dose in RA group was insignificant whereas significant in the subgroups of SA.Conclusions In comparison with conventional angiography,the dual-axis rotational angiography has the potential to reduce radiation exposure of patients significantly so as to prevent skin radiation injures.

Changes of cardiac function and left atrial diameter after radiofre-quency catheter ablation in patients with atrial fibrillation / 心血管康复医学杂志

Objective: To observe curative effect of radiofrequency catheter ablation (RFCA) and changes of cardiac function and left atrial diameter after operation in patients with atrial fibrillation (AF). Methods: A total of 28 AF patients with obvious clinical symptoms and without effective for drug therapy received RFCA in our hospital, their data were retrospectively analyzed. They received RFCA under guidance of three dimensional electro-anatomic mapping (EAM) system. Changes of cardiac function and left atrial diameter were evaluated by echocardiography and curative effect was evaluated by ambulary blood pressure monitoring before, three and six months after operation. Results: Pulmonary vein isolation rate was 100% in all patients. No severe complication occurred during or after operation. After six-month follow-up, 27 cases（96.4%）did not recur AF among the 28 patients; Compared with before operation, there were significant decrease in left atrial diameter [(37.3±4.8) mm vs. (33.6±4.5) mm] and significant increase in left ventricular ejection fraction [(59.8±8.7) % vs. (64.2±6.8) %] by echocardiography, P<0.05 both. Conclusion: Radiofrequency catheter ablation is safe and effective in treatment of atrial fibrillation, and there are significant improvements in cardiac function and left atrial diameter.

The clinical safety of dual axis rotational angiography in the diagnosis of coronary artery disease / 中华放射学杂志

Objective To observe the clinical safety of dual axis rotational coronary angiography (DARCA) in the diagnosis of coronary artery disease in Chinese population.Methods From March to December in 2010,74 patients undergoing diagnostic DARCA were enrolled.The improved isocentering technique was adopted in 34 of the patients at the end of the study during DARCA.Blood pressure,heart rate and symptoms were recorded immediately before-and-after contrast injections.Contrast dose,radiation exposure and procedure time for DARCA were recorded.Continuous variable data were analyzed using Student＇s t test,if normality assumption was violated,rank sum test would be used.Categorical variables were analyzed using x2 test.Results ( 1 ) Clinical safety:There was no chest pain documented during or immediately post-injection for all patients.Only 1 patient ( 1 ％ ) had an attack of ventricular tachycardia immediately after the contrast injection and then relieved automatically.Pre and post-injection systolic blood pressure values of left coronary artery were statistically different [ ( 116 ± 20 ) mm Hg vs.( 111 + 18) mm Hg( 1 mm Hg =0.133 kPa),t =3.303,P =0.001 ],and heart rates differed,too [ 73 ( 65- 84)bpm vs.71(64-78) bpm,Z =-4.789,P =0.001 ],but that imposed no clinical significance.(2)Contrast dose,radiation dose and procedure time:The mean contrast utilization,radiation dose and procedure time for DARCA were 28 (25-34) ml,8979 ( 6733-12 363 ) mGycm2 and 200 (164-270) s.Compared with conventional DARCA,improved isocentering technique during DARCA had less radiation exposure and procedure time in left coronary artery angiography and the whole coronary artery angiography [ left coronary artery angiographic radiation exposure:4004 (2932-5772) mGycm2 vs.5808 (4798- 8838) mGycm2,Z =-3.471,P =0.001 ;total radiation exposure:(8116 +2493) mGycm2 vs.( 11 371 ±4122) mGycm2,t =-4.176,P =0.001 ; left coronary artery angiographic procedure time:120 ( 80-180)s vs.150(126-214) s,Z =- 2.836,P =0.005; total procedure time:180 (139-240) s vs.220( 186-308 )s,Z =-3.004,P =0.003 ],but there was no statistically difference in contrast utilization [30(25-35) ml vs.27(25-34)ml,Z=-0.906,P=0.365].Conclusion This study demonstrates clinical safety of DARCA in the diagnosis of coronary artery disease in Chinese population.Compared with conventional isocentering technique of DARCA,improved isocentering technique can significantly reduce radiation exposure and procedure time on the basis of simplified operation,and replace the conventional isocentering technique,but randomized double-blind controlled studies should be conducted.

Safety and feasibility of Szabo technique in percutaneous coronary intervention of coronary artery ostial lesions / 心血管康复医学杂志

Background and Object: Percutaneous coronary intervention (PCI) on ostial lesions in coronary arteries associated with higher procedural difficulty and complication morbility. It has been technically difficult because it should be done with precise stent placement in ostium and absence of side branch compromise. The Szabo technique consists of side branch wiring through most proximal stent strut as well as main branch wiring through stent lumen. The side branch wire or anchor wire prevents stent advancement beyond ostial segment and makes possible the accurate stent implantation in ostium. The purpose of this study is to evaluate the safety, feasibility and success rate of Szabo technique by analysing technical, angiographic and IVUS (Intravascular Ultrasonography) findings. Methods: The data of 39 PCIs in 39 patients with a significant lesion at a coronary artery ostium which was treated percutaneously using Szabo technique were retrospectively analysed. The technically successful procedure was defined as there was neither stent loss nor second guide wire pull back during stent advancement. A successful procedure from angiographic point of view was defined as a precise stent implantation at ostium without side branch compromise. Successful procedure from IVUS point of view was defined as accurate stent placement in ostium without proximal protrusion and without any stent uncovered area. A total of 39 patients was with 28 males and 11 females. They aged from 43 to 79 years with a mean age of （65±12）years. The 6F and 7F guiding catheter were used in 35 (89.7%) and 4 (10.3%) patients separately. The access was radial in 31 (79.5%) and femoral in 8 (20.5%) patients. The culprit vessel was left anterior descending (LAD) in 26(66.7%), right coronary artery (RCA) 5 (12.8%), circumflex-obtuse marginal (LCX-OM) 3(7.7%), and posterior descending (PDA) 5(12.8%). IVUS was performed through culprit vessel in 30 (76.9%) and was also done in side branch in 9 (23.1%) patients after stent implantation. Results The procedure was technically successful in 36 (92.3%) patients. All technically successful patients had angiographic success (100%). IVUS examination of culprit vessel showed accurate stent placement in ostium 29 (96.7%) and slight stent proximal protrusion in 1(3.3%) patients. No serious complication occured. Conclusions This study shows that Szabo technique is safe and feasible for PCI in ostial coronary artery lesions with a high angiographic success rate，its deserved to spread.

Effects of Keratinocyte Growth Factor 2(KGF-2) on Keratinocyte Growth, Migration and on Excisional Wound Healing / 生物化学与生物物理进展

Keratinocyte growth factor 2 (KGF-2) is a member of the FGF family that is mainly synthesized by mesenchymal cells and acta predominantly on epithelial cells in a paracrine manner. It is known to play an important role in fetal limb and lung development; skin wound healing and prostatic epithelial cell growth. The KGF-2 coding sequence were isolated from human kidney cDNA library, revealing that the Kgf-2 gene is also expressed in the kidney apparatus. Purified from prokaryotic E. coli cells, the effects of the recombinant KGF-2 protein in cultured keratinocyte were analyzed by using MTT assay and in situ TUNEL assay. Interestingly, results revealed that KGF-2 promoted keratinocyte cell growth by stimulating cell proliferation and attenuating cell apoptosis. These findings supported a few evidences that KGF-2 could contribute to alveolar epithelial cells against apoptosis. Cell migration assays for the first time revealed that KGF-2 could stimulate keratinocyte cell migration in vitro. In addition, in the pilot animal test, recombinant KGF-2 incorporated within the hydrogel dressing exhibited significantly stimulatory effect on cutaneous wound healing. These combined effects implicate a potential therapeutic application of human recombinant KGF-2 in the future.

Effects of hypoxia-indudble factor 1α on the growth of subcutaneously implanted hepatocellular carcinoma in nude mice / 中华普通外科杂志

Objective To study the effects of HIF-1α expression on the growth of subcutaneously implanted human hepatocellular carcinoma (HCC) cell lines in nude mice. Methods A nude mouse model of subcutaneously implanted HCC cell line HepG2Tet-on-HIF-1α was established. This cell lines characterizes the inducible expression of HIF-1α by doxycycline (Dos). The impact of HIF-1α expression induced by Dox on the growth of subcutaneously implanted HCC cell lines in the nude mouse model was observed. Results The mRNA and protein expression of HIF-1α were significantly up-regulated in the nude mouse model by oral administration of Dox. Compared with that model in which Dox was not administrated ie Dos( - ) group, the tumor volume(513.545 ± 276. 229) mm3 vs. ( 166. 506 ± 110. 142) mm3 ( P < 0. 05 ), tumor weight ( 1.251 ± 0. 438 ) g vs. (0. 640 ± 0. 296) g ( P < 0. 05 ), and tumor growth velocity were significantly enhanced in Dox ( + ) group, while tumor necrosis was inhibited ( 31. 360% ±2. 728% vs. 36. 640% ± 3. 804% ) (P<0. 05). The weight loss of nude mice was larger in Dox( + )group( P < 0. 01 ). There was no liver or lung metastasis in either group. Conclusion The expression of HIF-1αin subcutaneously implanted HCC in a nude mouse model is up-regulated by oral Dox. High grade expression of HIF-1α promotes the growth of implanted HCC.

The expression of livin and hepatitis B virus X protein in HCC / 中华普通外科杂志

Objective To investigate the expression of livin in hepatocarcinoma tissues and its relationship with hepatitis B virus X protein. Methods Immunohistochemistry was used to detect the expression of livin in 30 cases of hepatic carcinoma tissues and 10 cases of normal liver tissues. Stable L02 cell hne tansfected with HBx was established. Real-time PCR and Western blot were used to observe the expression of livin gene before and after the HBx transfeetion. Results The protein expression of livin was higher in hepatocarcinoma tissues than in normal tissues, and correlated with the exist of portal vein cancer embolus (t = 2.24, P = 0.033 ; P<0.05), but it was not correlated with cancer size or classification pathologically(t = 1.688, P = 0.103 ; γ = - 0.137, P = 0.471 ; P＞0.05). Livin Mrna expression was up-regulated after the HBx gene transfection (t = 6.73, P<0.05). Conclusions Apoptosis inhibition induced by livin up-regulation may play a role in pathogenesis and development of hepatocarcinoma. HBx up-regulates livin expression at transcriptional level.

A modified simple RFLP-PCR method for single nucleotide polymorphism (SNP) typing

We describe a modified single nucleotide polymorphism (SNP) typing method based on the restriction fragment length polymorphism polymerase chain reaction (RFLP-PCR). This is a simple, economical method without the need for special equipment. For most SNP loci, a common restriction endonuclease (Hind III, EcoR I or BamH I) recognizing site (RER) can be introduced into one allelic form, but not the other by two rounds of mismatched PCR. The flanking regions can be changed by as many as five bases after PCR amplification with specially designed mismatching primers so the genotypes can be distinguished after digestion of the PCR products with corresponding endonucleases.

The establishment of the modified cultural technique of cardiomyocytes in human fetal hearts / 医学研究生学报

Objectives:This study was designed to establish the modified method of cardiomyocytes culture in human fetal hearts. Methods:The human fetal heart cells of ventricular muscle were isolated by 0.2% trypsin and 0.1% collagenase and cultured primarily and passaged in Iscove's modified Dulbecco's medium(IMDM) in vitro by means of differential attachment technique.The changes in morphology,ultrastructure,viability,immunocytochemistry antibody staining and immunofluorescence antibody staining of human fetal heart cells were studied in culture. Results:The ratio of viable cells was 99% identified by trypan blue staining.The ratio of attachment cells was 95% after 24 h in culture.The cultured human fetal heart cells were roundness shaped,rod shaped,shuttle shaped,ellipse shaped,star shaped and bifurcate shaped with spontaneous contractility.The myocardial actin and myoglobin are identified in the cultured cells by immunocytochemistry antibody staining and immunofluorescence antibody staining.The ultrastructure of cells was similar to that of the cardiac tissue in vivo by electron microscopy.Human fetal heart cells after 20 d of primary culturing and after 5 d of passaged culturing were growed well. Conclusions:The method for isolating and culturing human fetal heart cells is successful and reliable.This model provides an effective experimental mothod for studying the mechanism of myocardial injury.

Specific anti-tumor immune response induced by pEGFP-C3-B7.2-MAGE-1 fusion DNA vaccine / 中国免疫学杂志

Objective:To construct pEGFP-C3-B7.2-MAGE-1 eukaryotic expression plasmids and to study the immunological effects of esophageal cancer cell vaccine modified by human B7.2/MAGE-1 gene.Methods:The B7.2 and MAGE-1 cDNA,which was amplified by RT-PCR, and the eukaryotic expression plasmid pEGFP-C3-B7.2-MAGE-1 was constructed. Human esophageal cancer cell EC9706 was transfected with the vector of pEGFP-C3-B7.2-MAGE-1 using the technique of lipofectamine transfection.The dendritic cells(DCs)from peripheral blood mononuclear cells(PBMC)were loaded with the tumor antigen,and co-cultured with congeneric T cells derived from PBMCs for 3 days to obtain the tumor specific cytotoxicity T lymphocytes(CTL). Methy1 thiazoly1 tetrazolium (MTT) assay was used to detect inhibition effect of CTL on transfected and untransfected EC9706 cells.Results:The CTL had stronger inhibition response against the cancer cells transfected with pEGFP-C3-B7.2-MAGE-1 than to the cancer cells transfected with pEGFP-C3 and the untransfected cancer cells(P