1.Construction and identification of eukaryotic expressed plasmid vector pLenti6/V5- DEST-Cx43 in human
Huimin FAN ; Yang LI ; Zhongmin LIU
Chinese Journal of Thoracic and Cardiovascular Surgery 2003;0(02):-
Objective To construct eukaryotic expressed plasmid vector pLenti6/V5- DEST-Cx43 in human for further study on bioinformative communication between myocardial cell, embryonic development of heart and differentiation of myocardial cell. Methods Cx43 cDNA fragments were obtained from plasmid p18-T by restriction enzyme BamH1、Xho1, after the agarose gel electrophoresis was performed, the Cx43 cDNA was retrieved. pENTR11 were extracted by enzyme BamH1、Xho1, then connected with Cx43 cDNA by T4 DNA ligase. LR reaction was performed and Cx43 cDNA were cloned into Lentiviral vector pLenti6/V5-DEST. Results Agarose electrophoresis and sequent examination, identified that Cx43 cDNA was cloned into Lentiviral vector pLenti6/V5-DEST. Conclusion The present experiment demonstrate a successful cloning of human Cx43 cDNA into the Lentiviral vector pLenti6/V5-DEST.
2.Effect of lentivirus expressing TLR4 shRNA on function of rat alveolar macrophage
Jian LIU ; Huimin FAN ; Zhongmin LIU
Academic Journal of Second Military Medical University 1985;0(05):-
Objective To construct the replication-incompetent lentivirus expressing rat TLR4 shRNA,and to observe its inhibitory effect on TLR4 expression in alveolar macrophage and on lipopolysaccharide (LPS)-induced release of IL-1?,IL-6.Methods We designed and constructed four shRNA expressing plasmids with silencing effect,then they were co-transfected into HEK293T cells with the TLR4 eukaryotic vector constructed previously;the best shRNA was selected to be packed into lentivirus;and the titer was determined.The packed lentvirus was used to infect the alveolar macrophage(NR8383) in presence of LPS,and the LPS-induced IL1?,IL-6 expression was examined by ELISA.Results The best shRNA was successfully screened out and was correctly inserted into the lentivirus.The titer of the recombinant lentivirus was 2.0 ? 106 TU/ml.The LPS-induced expression of IL-1?,IL-6 in the alveolar macrophages was greatly reduced after virus infection (P
3.Updated biological characterization of Th17 cells
Hao CAO ; Huimin FAN ; Zhongmin LIU
Journal of Medical Postgraduates 2003;0(11):-
Th17 cells play an important role in the pathogenesis of various immune-mediated diseases,including psoriasis,rheumatoid arthritis,multiple sclerosis,inflammatory bowel disease,asthma,and so on.The discovery of Th17 cells has offered scientists a new insight into the etiology and treatment of a broad spectrum of diseases.This article presents an overview of the differentiation,cytokine expression and trafficking of Th17 cells.
4.Heart-lung transplantation for 4 patients with Eisenmenger’s syndrome
Huimin FAN ; Zhongmin LIU ; Jianhua GUO
Chinese Journal of Organ Transplantation 2003;0(05):-
Objective To summarize the experience of heart-lung transplantation.Methods Four patients with Eisenmenger’s syndrome underwent heart-lung homoplastic transplantation. All patients were complicated with severe pulmonary hypertension in New York Heart Association ( NYHA ) functional class IV. Cannulation for cardiopulmonary bypass consisted of a cannula in the high ascending aorta and separate vena caval cannulas. The heart-lung graft was moved into the chest, beginning with passage of the lung before the phrenic nerve pedicle. The bronchus was trimmed, leaving two cartilaginous rings proximal to the orifice of the upper lobe. The tracheal anastomosis was performed with a continuous 4-0 polypropylene suture, with the posterior portion continuously and anterior interrupted. The lungs were then ventilated (
5.Combined procedure of surgical treatment and cell transplantation for dilated cardiomyopathy: initial results
Huimin FAN ; Rong LU ; Liang MA ; Yang LI ; Zhongmin LIU
Chinese Journal of Thoracic and Cardiovascular Surgery 2008;24(2):95-97
Objective This study evaluated the effects of combined procedure of surgical treatment and cell transplantation on dilated cardiomyopathy. Methods Eight patients (5 men and 3 women) with moderate to severe mitral regurgitation from end-stage dilated cardiomyopathy underwent surgery. Four patients were in functional class (FC) Ⅳ, six were in FC Ⅲ. There age ranged from 15 to 56 years. The preoperative ejection fraction (EF) ranged from 0.15 to 0.32 (mean 0.26 ± 0.08). Mitral valve replacement was performed in 5 patients and mitral valve repair in 3. Auto-bone marrow monenuclear cells were harvested, isolated, washed, and resuspended for direct injection after surgical procedure. Results All patients survived and were discharged from the hospital. After a mean follow-up period of 18 months ( 12 - 42 months). Echocardiography showed postoperative ejection fraction and wall movement velocity increased after 6 months. Radionuclide ventriculography showed myocardial perfusion improved significantly. Conclusion Combined procedure of surgical treatment and cell transplantation led to significant improvement in cardiac function in patients with dilated cardiomyopathy.
6.A case of blindness caused by nephritic syndrome with cerebral venous sinus thrombosis
Tao SUN ; Zhengkun XIA ; Zhongmin FAN ; Zhuo SHI ; Yuanfu GAO
Journal of Clinical Pediatrics 2014;(2):160-163
Objectives To explore the clinical manifestations, treatment and prognosis of a case of blindness caused by nephrotic syndrome with cerebral venous sinus thrombosis (CVST). Methods The clinical manifestations, diagnosis and treatment of a case of NS with CVST were analyzed. The latest domestic and foreign reseach progresses in treatment for CVST in children were reviewed. Results Epilepsy suddenly appeared with diplopia, binocular vision loss and blindness in anticoagulant therapy for the child with NS. Brain magnetic resonance venography (MRV) suggested CVST. MRV reexam-ined showed that the intracranial thrombosis was completely dissolved after urokinase thrombolysis for one month followed by ineffective heparin anticoagulation. At present, international standards of anticoagulant therapy have been adopted in the treatment for CVST patients. Coagulation function (e.g.APTT) and international standardization ratio were monitored in order to prevent bleeding. Conclusions It is better to perform neural imaging examination early in suspected CVST patients. Anti-coagulation and thrombolytic therapy should be given immediately once the risk of bleeding was excluded and used for 3-6 months.
7.Inflammatory Factors and von Willebrand Factor in Hypertensive Patients Complicated with Metabolic Syndrome
Jing LIU ; Zhongmin TANG ; Qian FAN ; Jishun MA ; Ningling SUN
Chinese Journal of Hypertension 2006;0(10):-
Objective To test the hypothese that metabolic syndrome is closely associated with inflammation, we compare the difference of inflammatory factors and von Willebrand factor between essential hypertensives and hypertensive patients with metabolic syndrome. Methods According to the new definition of metabolic syndrome by IDF 2005, one hundred thirty eight consecutive hospitalized essential hypertensives were catelogorized into EH+MS group, n=99 and EH alone group, n=39. Biochemistry assay, white blood cell (WBC) counts, serum high sensitive C reactive protein (hsCRP) and plasma von Willebrand factor (vWF) were measured. The relationship between hsCRP, vWF and MS were analysed. Results Waist circumference, body mass index, triglyceride, low density lipoprotein cholesterol (LDL-C), WBC, serum hsCRP and plasma vWF were increased(P
8.The strategy of diagnosis and treatment of anti-neutrophil cytoplasm antibody-associated vasculitis
Zhengkun XIA ; Qingshan ZHOU ; Yuanfu GAO ; Zhongmin FAN
Journal of Clinical Pediatrics 2016;34(6):401-405
Anti-neutrophil cytoplasm antibody-associated vasculitis (ANCA) is an autoimmune disease with multi organ involvement characterized by vascular wall inflammation and fibrinoid necrosis, including microscopic polyangitis (MPA), granuloma polyangitis (GPA), and eosinophilic granuloma polyangitis (EGPA). Because its clinical manifestations are complicated and non-speciifc, it is dififcult to make early diagnose. In recent years, some new progress has been made in diagnosis and treatment of this disease. The article will review the related information.
9.The hemolysis research and fatigue test in vitro of the national implantable ventricular assist device
Albahde MUGAHED ; Liang YE ; Huimin FAN ; Zhongmin LIU
Chinese Journal of Thoracic and Cardiovascular Surgery 2016;32(1):34-36
Objective To evaluate the hemolysis and fatigue resistance self suspension ventricular assist device (NIVAD) in vitro.Methods Resettled NIVAD which is filled with 2 500 ml fresh anticoagulant blood from pig in vitro loop system.The speed of blood pump was adjusted to 8 000 rpm.The afterload and the flow of pump were set as 100 mmHg and 3.5 L/min respectively.Keep the temperature at around 37 degree centigrade which is equal to average internal temperature in pig.Then harvest and measure the free hemoglobin(FHB) of blood samples at different time points before and after working.Then calculated the normalized index of hemoylsis(NIH) by comparing to control group, the unused pig blood at the same period.In fatigue test, conditions remained unchanged, except the blood was replaced by pure water.The blood pump had operated for 90 d continuously and recorded its performance.Results The FHB of experimental blood is higher than that of the control.The normal index of hemolysis NIVAD is(0.0052 ±0.0019)mg/L.Fatigue test , the blood pump operated smoothly, and every parameters was stable.Conclusion The blood pump of NIVAD does a certain degree of destruction to blood, but the NIH is in criterion range of hemolysis, it has certain advantages compared to similar products in china and aboard.At the same time it has good resistance and stable property to fatigue.
10.Novel deletion mutation of type-Ⅳcollagen in a Chinese family with Alport syndrome
Chunlin GAO ; Zhengkun XIA ; Zhongmin FAN ; Yuanfu GAO
Journal of Medical Postgraduates 2015;(9):929-933
Objective Alport syndrome is one of the diseases that may lead to the end-stage renal disease ( ESRD) in chil-dren, and the methods for its diagnosis and treatment remain quite limited.This study aimed to investigate the clinical and genetic di-agnosis of a Chinese family with hematuria companied by genetic nephritis. Methods We analyzed the renal pathology of 7 patients in a family, performed immunofluorescence staining of type-Ⅳcollagen in the nephridial and skin tissue, conducted gene sequencing i-dentification using the exon sequence method, and examined the blood and urine samples from the patients. Results Renal patholo-gy manifested mesenterium hyperplasia in the index patient, with IgM+under the light microscope, no thickening or thinning under the electromicroscope, and no absence of type-Ⅳcollagen on immunofluorescence analysis.Mutation of c.1365_1373del TCCAGGCCC (p.Pro456_Pro458del3) was observed in exon 21 of the COL4A5 gene.Only 1682 amino acids were found in the mutated protein as compared with 1685 in the wild type. Conclusion This is the first case of Alport syndrome induced by gene deletion mutation ever reported in China and abroad.There are many female patients in this family, all with a high risk of reproduction failure.Antepartal gene diagnosis or genetic diagnosis before embryo transfer may contribute to the prevention of the disease.