Gene mutation profiling and clinical significances in patients with renal cell carcinoma

Abstract Objectives The pathological mechanisms of patients with Renal Cell Carcinoma (RCC) remain defined. This study aimed to evaluate relationships between the landscape of gene mutations and their clinical significance in RCC patients. Methods Tissue and peripheral blood samples of 42 patients with RCC were collected and performed for the Next Generation Sequencing (NGS) with Geneseeq PrimeTM 425-gene panel probes. Their landscapes of gene mutation were analyzed. We also carried out an evaluation of Tumor-Node-Metastasis (TNM) staging, RENAL nephelometry score, surgery, and targeted drug treatment of patients. Then we compared the correlations of landscape in gene mutations and the prognosis. Results The most common gene alternations, including BAP1, PBRM1, SETD2, CSF1R, NPM1, EGFR, POLE, RB1, and VHL genes, were identified in tissue and blood samples of 75% of patients. EGFR, POLE, and RB1 gene mutations frequently occurred in relapsed and metastatic patients. BAP1, CCND2, KRAS, PTPN11, ERBB2/3, JAK2, and POLE were presented in the patients with > 9 RENAL nephelometry score. Univariable analysis indicated that SETD2, BAP1, and PBRM1 genes were key factors for Disease-Free Survival (DFS). Multivariable analysis confirmed that mutated SETD1, NPM1, and CSF1R were critical factors for the Progression Free Survival (PFS) of RCC patients with target therapy. Conclusions Wild-type PBRM1 and mutated BAP1 in patients with RCC were strongly associated with the outcomes of the patient. The PFS of the patients with SETD2, NPM1, and CSF1R mutations were significantly shorter than those patients without variants.

Identification of activated pathways in lung adenocarcinoma based on network strategy

Background: Lung adenocarcinoma has increased incidence over the past years and is the cause for almost 50% of deaths attributable to lung cancer. The objective of this paper is to identify activated pathways associated with lung adenocarcinoma based on gene co-expression network analysis. Materials and Methods: Kyoto Encyclopedia of Genes and Genomes pathway analysis of dysregulated genes was performed based on Expression Analysis Systematic Explorer test to illuminate the biological pathways. Co-expression networks of lung adenocarcinoma in different tumor Stages (IA, IB, IIA, IIB, IIIA, IIIB, and IV) were constructed by Empirical Bayes approach to reweight gene pair scores. Pathway activity analysis was conducted to compute the distribution of pathways in different stages and to identify “activated” pathways in lung adenocarcinoma. Results: We evaluated 211 dysregulated genes between lung adenocarcinoma patients and normal controls. Pathway activity analysis was performed and P values of pathways, which obtained from co-expression networks (Stage IA, IB, IIA, IIB, IIIA, IIIB, and IV), were calculated. Cell cycle, progesterone-mediated oocyte maturation, and oocyte meiosis were activated during all stages in lung adenocarcinoma. Conclusions: We successfully identified three activated pathways (cell cycle, progesterone-mediated oocyte maturation, and oocyte meiosis) in different Stages (IA, IB, IIA, IIB, IIIA, IIIB, and IV) of lung adenocarcinoma

Preliminary study of the relationship between novel coronavirus pneumonia and liver function damage: a multicenter study / 中华肝脏病杂志

To analyze the clinical characteristics of cases of novel coronavirus pneumonia and a preliminary study to explore the relationship between different clinical classification and liver damage. Consecutively confirmed novel coronavirus infection cases admitted to seven designated hospitals during January 23, 2020 to February 8, 2020 were included. Clinical classification (mild, moderate, severe, and critical) was carried out according to the diagnosis and treatment program of novel coronavirus pneumonia (Trial Fifth Edition) issued by the National Health Commission. The research data were analyzed using SPSS19.0 statistical software. Quantitative data were expressed as median (interquartile range), and qualitative data were expressed as frequency and rate. 32 confirmed cases that met the inclusion criteria were included. 28 cases were of mild or moderate type (87.50%), and four cases (12.50%) of severe or critical type. Four cases (12.5%) were combined with one underlying disease (bronchial asthma, coronary heart disease, malignant tumor, chronic kidney disease), and one case (3.13%) was simultaneously combined with high blood pressure and malignant tumor. The results of laboratory examination showed that the alanine aminotransferase (ALT), aspartate aminotransferase (AST), albumin (ALB), and total bilirubin (TBil) for entire cohort were 26.98 (16.88 ~ 46.09) U/L and 24.75 (18.71 ~ 31.79) U/L, 39.00 (36.20 ~ 44.20) g/L and 16.40 (11.34- ~ 21.15) mmol/L, respectively. ALT, AST, ALB and TBil of the mild or moderate subgroups were 22.75 (16.31- ~ 37.25) U/L, 23.63 (18.71 ~ 26.50) U/L, 39.70 (36.50 ~ 46.10) g/L, and 15.95 (11.34 ~ 20.83) mmol/L, respectively. ALT, AST, ALB and TBil of the severe or critical subgroups were 60.25 (40.88 ~ 68.90) U/L, 37.00 (20.88 ~ 64.45) U/L, 35.75 (28.68 ~ 42.00) g/L, and 20.50 (11.28 ~ 25.00) mmol/L, respectively. The results of this multicenter retrospective study suggests that novel coronavirus pneumonia combined with liver damage is more likely to be caused by adverse drug reactions and systemic inflammation in severe patients receiving medical treatment. Therefore, liver function monitoring and evaluation should be strengthened during the treatment of such patients.

Comparative study of surgical and interventional closure for the treatment of patent ductus arteriosus in children / 中国胸心血管外科临床杂志

@#Objective    To study effect of different surgical treatments for patent ductus arteriosus in children. Method    A total of 38 patients with patent ductus arteriosus who underwent surgical treatment of cardiothoracic surgery between January 2016 and December 2017 in our hospital were as an observation group (12 patients with severing suture, 26 patients with ligation, 14 males and 24 females, aged 0.08–8.67 years). In the same period, 38 patients with patent ductus arteriosus who underwent interventional closure in the Department of Cardiology were as a control group (17 males and 21 females, aged 0.50–5.42 years). The clinical effectiveness of the two groups was compared. Results    The operation time, postoperative hospital stay, and blood transfusion rate in the observation group were higher than those in the control group with statistical differences (P<0.05). There was no statistical difference in intraoperative blood loss and complications between the two groups. Conclusions    In patients with a single patent ductus arteriosus or a small catheter, interventional closure of the patent ductus arteriosus is less trauma and faster recovery. But in patients with lower weigh, premature, other intracardiac malformations, large catheter, moderate or severe pulmonary hypertension, the surgery is better.

Comparison of Apical and Asymmetric Septal Phenotype in Patients With Hypertrophic Cardiomyopathy: Clinical Features and Prognosis / 中国循环杂志

Objectives: To compare the clinical features and long-term outcomes of patients with apical hypertrophic cardiomyopathy (ApHCM) and patients with asymmetric septal hypertrophic cardiomyopathy (ASHCM). Methods: Data from 600 patients (300 with ApHCM and 300 with ASHCM) identified in a consecutive single-center cohort between 1996 and 2014 were retrospectively analyzed. The two groups were 1:1 matched by age of diagnosis, gender and the presence of outflow tract obstruction. Clinical features, cardiovascular mortalities, incidence of sudden cardiac death and cardiovascular morbidity (including unexplained syncope, atrial fibrillation, nonsustained ventricular tachycardia, progressive heart failure, embolic stroke or transient ischemic attack and myocardial infarction) were compared between the two groups. Results: Forty-two patients (14.0%) had a maximum LV wall thickness of ≥30 mm in the ASHCM group compared to only 11 patients (3.7%) in the ApHCM group (P<0.01). 156 patients in ApHCM group (52.0%)and 168 patients in ASHCM group(56.0%)underwent cardiovascular NMR examination, the incidence of late gadolinium enhancement was significantly lower in ApHCM group than in ASHCM group(26.9% vs 76.2%,P<0.01). The mean follow-up durations for ApHCM and ASHCM were (7.5 ± 4.0) years and (6.6 ± 5.4) years, respectively. The incidence of cardiovascular death (1.0% vs 5.7%), sudden cardiac death (0.33% vs 3.3%) and major adverse cardiovascular event (18.3% vs 40.3%) were significantly lower in the ApHCM group than in the ASHCM group (all P<0.01). Unexplained syncope, nonsustained ventricular tachycardia, and progressive heart failure were less common in ApHCM group than in ASHCM group (all P<0.05). Multivariate COX regression analysis showed that late gadolinium enhancement positivity (HR=4.62, 95% CI: 2.28- 68.0, P=0.02) and unexplained syncope (HR=8.56, 95% CI: 2.1-16.6, P<0.01) were independent predictors of cardiovascular mortality. Unexplained syncope was independent predictor for sudden cardiac death (HR=4.40, 95% CI: 1.5-15.2, P=0.02). Conclusions: After eliminating the interference of age at diagnosis, gender and outflow tract obstruction, patients with ApHCM represent a more benign prognosis with a lower incidence of cardiovascular mortality and morbidity than patients with ASHCM.

Efficacy of Extended Myectomy on Reducing Mitral Regurgitation in Patients With Hypertrophic Obstructive Cardiomyopathy / 中国循环杂志

Objectives:To analyze the impact of extended myectomy on reducing mitral regurgitation in patients with hypertrophic obstructive cardiomyopathy (HOCM). Methods:We retrospectively analyzed 480 consecutive HOCM patients who underwent surgical treatment by the same surgeon in our institution from October 2002 to July 2017. The efficacy of extended myectomy for reducing mitral regurgitation and left ventricular outflow tract (LVOT) obstruction were evaluated by echocardiography after surgery. Results:Among the 480 patients, 22 (4.6%) received concomitant mitral repair or replacement because of their intrinsic mitral diseases. In the remaining 458 (95.4%) patients without concomitant mitral valve surgery, 1 (0.2%) died at the 5th day after surgery because of infective shock, and another 4 (0.9%) lost to follow-up, a total of 453 (98.9%) patients underwent echocardiographic follow-up (median follow-up time:6 months [3, 12]). During follow-up, left ventricular out flow tract gradient was significantly decreased from (89.1±30.6) to (12.8±11.6) mmHg (P＜0.001); the number of patients with systolic anterior motion (SAM) of mitral leaflets decreased from 451(98.5%) to 42 (9.3%) (P＜0.001); 297 (64.8%) patients presented with moderate or severe mitral regurgitation before surgery, which decreased to 14 (3.1%) at follow-up (P＜0.001); the multivariate regression analysis showed that patients with residual SAM were significantly associated with a higher incidence of moderate to severe mitral regurgitation during follow-up (odds ratio 30.334, 95% confidence interval:5.619-163.739, P＜0.001). Conclusions:Extended myectomy, combined with dividing the anomalous links between mitral apparatus and septum, and trimming papillary muscles, yields satisfactory outcomes of relieving LVOT obstruction and reducing mitral regurgitation in most of patients with HOCM. Concomitant mitral valve surgery is rarely required unless the patient have intrinsic mitral valve disease.

2011 Status Survey for Physical Activity and Regular Exercise in Adult Residents of Beijing / 中国循环杂志

Objective: To explore the physical activity (PA) and regular exercise in adult residents of Beijing in order to provide the basis for guiding regular exercise and chronic disease prevention. Methods: A multi stage, random cluster investigation was conducted in 20242 residents from (18-79) years in Beijing area. Based on International physical activity questionnaire (IPAQ) published in USA (2000), the metabolic equivalent (MET) was calculated; according to the standard by General administration of sport of China, regular exercise rate was assessed. Weighting adjustment was conducted on the basis of 2010 Beijing population census data and sampling design. Results: 19584 participants with complete information were enrolled for final analyses. The median weekly MET of occupational PA, domesticity, transport-related PA and leisure exercise were (1918, 839, 922 and 928) MET-minute/week respectively. Female had the lower MET in occupational PA and leisure exercise than male (1436 vs 2156) MET-minute/week and (827 vs 944) MET-minute/week, both P<0.001. Elderly adults had the higher MET in leisure exercise than young and middle aged adults (1490 vs 718) MET-minute/week and (1490 vs 1162) MET-minute/week, both P<0.001. Regular exercise rate was only 31.5％ (95％ CI 30.7％-32.4％); multivariate analysis indicated that compared with male, female had the higher risk of insufficient regular exercise, OR=1.26 (95％ CI 1.14-1.40), compared with young adults, middle and elderly adults had the lower risk of insufficient regular exercise, OR=0.54 (95％ CI 0.50-0.59) and OR=0.46 (95％ CI 0.39-0.54). Compared to residents with education≤6 years, those with education at (7-12) years and with education>12 years had the lower risk of insufficient regular exercise, OR=0.66 (95％ CI 0.57-0.77) and OR=0.46 (95％ CI 0.39-0.54). Conclusion: Sampling residents in Beijing had insufficient leisure exercise with the low rate of regular exercise in 2011. Health policy and environment especially beneficial to female, youth and lower educated adults should be established to improve the healthy Beijing construction.

Expression dynamics of caveolin-1 in fibroblasts of newborn rats with chronic lung disease and its impact on lung fibroblast proliferation

Abstract Purpose: To evaluate the changes of caveolin-1 in lung fibroblasts in newborn Wistar rats when exposed to hyperoxic conditions, as well as lung fibroblasts cell cycle. Methods: One hundred newborn Wistar rats were randomly divided (50 rats/group) into experimental and control groups, exposed to hyperoxic conditions or normal air, respectively. The fraction of inspired oxygen (FiO2) in the experimental group was 90%, whereas this value was 21% in the control group. Lung fibroblasts were collected on days 3, 7, and 14 of the experiment. Caveolin-1 expression dynamics in lung fibroblasts was assayed in each group by immunofluorescence and Western blot analyses. Flow cytometry (FCM) was used to assess the proportions of lung fibroblasts at different stages of the cell cycle. Results: On day 3, no significant difference in caveolin-1 expression was observed between the hyperoxic and control groups; however, on days 7 and 14, caveolin-1 expression was significantly lower in the hyperoxic group than in the control (P<0.05). No apparent differences were observed in caveolin-1 expression in the control group at the different time points. Using FCM analysis, we showed that the proportion of lung fibroblasts in G0/G1 phase in the hyperoxic group decreased compared to that of the control group on day 7, while the proportion of S-phase cells increased (P<0.05). These differences were more significant when the groups were compared on day 14 (P<0.01). Conclusion: After seven days the exposure to hyperoxic conditions, lung fibroblasts proliferated and caveolin-1 expression decreased.

Clinical distribution and resistance analysis of pseudomonas aeruginosa in the four hospitals of Henan province during 2012-2015 / 新乡医学院学报

Objective To investigate the clinical distribution and resistance change of pseudomonas aeruginosa (PAE) in the four hospitals of Henan province during 2012-2015,so as to provide guidance for reasonable clinical use of antibiotics.Methods By means of disk diffusion method (K-B method) or automatic instrument,the drug susceptibility testing was performed for the clinical isolated strain which collected from the 371st Central Hospital of PLA,the Third Affiliated Hospital of Xinxiang Medical University,the 150th Central Hospital of PLA and the 153rd Central Hospital of PLA during 2012-2015,then the result were interpreted according to the criteria of Clinical and Laboratory Standards Institute 2010,and the date were analyzed by using WHONET 5.6 software.Results A total of 1 796 strains of PAE had been isolated,and mainly distributed in department of critical care medicine (32.6％) and department of gastroenterology (28.7％).The specimens mainly collected from sputum (74.6％) and wound secretion (9.6％).The drug susceptibility results showed that the antibiotic sensitive rate of PAE to polymyxin B(93.5％) was the highest,followed by the piperacillin/tazobactam(78.2％) and amikacin(76.2％).Resistance rate of PAE to piperacillin(34.4％) was the highest,followed by the ticarcillin/calvulanic acid(34.1％) and aztreonam (30.7 ％).Drug-resistance of PAE to the ceftazidime,cefepime,aztreonam,amikacin,gentamicin and polymyxin B had a downward trend,respectively.Conclusion Drug-resistance of the PAE to the most commonly used antimicrobial has a downward trend.However,the drug-resistance situation of PAE is very serious and the resistance rates still keep the high level during 2012-2015.It is necessary to investigate the clinical distribution and resistance of PAE,so as to provide guidance for reasonable clinical use of antibiotics.

Evaluation of left atrial function and its determinants by three-dimensional echocardiography in patients with hypertrophic cardiomyopathy / 中华医学超声杂志（电子版）

Objective To evaluate the left atrial function and to explore its determinants in patients with hypertrophic cardiomyopathy by three-dimensional echocardiography (3DE).Methods 46 patients with HCM (obstructive HCM:25 cases,nonobstructive HCM:21 cases) and 46 healthy cases (controls) were enrolled in this study.Time-volume curve of left atrium was acquired by 3DE in all subjects.Left atrial maximal volume (LAVmax),left atrial minimal volume (LAVmin) and left atrial presystolic volume (LAVp) were acquaired.Left atrial volume index (LAVI),left atrial expansion index (LAEI),left atrium emptying fraction (LAEF),left atrium passive emptying fraction (LAPEF) and 1eft atrium active emptying fraction (LAAEF) were calculated.Comparative analysis between two groups was taken .The Spearman correlation analysis and multiple linear regression analysis between left atrial volume index (LAVI) with interventricular septal thickness (IVSd),left ventricular outflow tract peak gradient (LVOT-PG),mitral regurgitation (MI), left ventricular diastolic function (LVDF) were analyzed respectively .Results Compared to the controls LAVmax (45.67 ±11.96)ml,LAVmin (20.48 ±6.80)ml,LAVp (24.48 ±9.31)ml,LAVI 25.63 ±6.52, LAEI (1.32 ±0.49)%,LAEF (55.25 ±8.06)%,LAPEF (35.90 ±7.00)%and LAAEF (30.20 ±10.13)%, the patient with HCM had a significantly larger LAVmax (81.45 ±24.24)ml,LAVmin (44.60 ±18.96)ml, LAVp (61.00 ±21.64) ml and LAVI 45.39 ±14.17,there were significant differences among the groups (t=8.978,8.123,9.227,8.436,all P0.01).There were significant positive correlation between LAVI and IVSd,LVOT-PG,MI,LVDF respectively (r=0.704,0.517,0.640,0.701,all P<0.01).Multiple linear regression analysis demonstrated that IVSd , LVOT-PG, MI and LVDF were correlated factors of LAVI (absolute standardized coeffients =0.264,0.515,0.614,0.341,all P<0.05).Conclusions 3DE could evaluate the left atrial volume and function in patients with HCM , with increased left atrial volume and decreased reservioer,conduit and booster pump function .Mitral regurgitation,obstruction of left ventricular outflow tract,left ventricular diastolic dysfunction and the thickness of left ventricular wall contributed to left atrial dysfunction at different levels ,among which mitrial regurgitation contributed the most .

Comparison of dobutamine stress echocardiography and exercise echocardiography in evaluating left ventricular outflow tract obstruction in patients with hypertrophic cardiomyopathy / 中华心血管病杂志

<p><b>OBJECTIVE</b>To compare left ventricular outflow tract (LVOT) gradient induced by dobutamine stress echocardiography (DSE) and exercise echocardiography (EE) in patients with hypertrophic cardiomyopathy (HCM).</p><p><b>METHODS</b>DSE and EE were performed in 40 consecutive patients with HCM and LVOT gradient < 50 mm Hg (1 mm Hg = 0.133 kPa) at rest. Dobutamine was administered intravenously at incremental doses of 200, 400, 600, 800 and 1200 µg/min at 5 min intervals. LVOT gradients were measured at rest, at peak exercise and during each dose of DSE.</p><p><b>RESULTS</b>LVOT gradient at rest was lower than 30 mm Hg in 36 patients and between 30 and 49 mm Hg in 4 patients. Of the 36 patients with LVOT gradient < 30 mm Hg at rest, mechanical LVOT obstruction (latent LVOTO) was evidenced in 17 patients during EE and in 18 patients during DSE and good consistency (91.7%, kappa value 0.833) was found between the two maneuvers. The provoked gradient was similar between DSE at 800 µg/min and EE at peak exercise (P = 0.181). In the 4 patients with LVOT gradient between 30 and 49 mm Hg [(38.8 ± 2.6) mm Hg], LVOT gradient increased to (85.3 ± 26.4) mm Hg during EE and (105.0 ± 28.0) mm Hg during DES.</p><p><b>CONCLUSIONS</b>DES and EE are comparable and suitable provoke methods for identifying LVOT obstruction in patients with HCM.</p>

Absorption and excretion of fluoride and aluminum after drinking brick tea in healthy adults / 中国地方病学杂志

Objective To observe the absorption, excretion and retention of fluoride and aluminum after drinking brick tea in healthy adults. Methods The study was conducted in ten healthy volunteers by drinking brick tea solution in which the fluoride and aluminum concentrations were 5.97, 7.53 mg/L, respectively. The concentrations of fluoride and aluminum were determined in serum samples collected before and at 0.5,1.0,2.0,3.0,and 24.0 h, and in urine samples collected before and during the periods 1.0,2.0,4.0, ＞ 4.0 - ＜ 24.0 h and 24 h after drinking brick tea solution by ion-selective electrode, inductively coupled plasma atomic emission spectrometry (ICP-AES) and inductively coupled plasma atomic mess spectrometry. The total amounts of intake and excretion of fluoride and aluminum in healthy volunteers during 24.0 h were calculated from their corresponding fluoride and aluminum ingesting from brick tea and excreting from urine. Results Before and during the periods 1.0,2.0,4.0,＞ 4.0 - ＜ 24.0 h and 24.0 h after drinking brick tea solution, the urinary fluoride concentrations were (0.50 ±0.14), (2.14 ± 0.90), (1.57 ± 0.93), (2.43 ± 1.49), (1.91 ± 0.69), (0.58 ± 0.20)mg/L, respectively, and the aluminum concentrations in the urine samples were (0.35 ± 0.15), (0.65 ± 0.37), (0.50 ± 0.25), (0.52 ± 0.21 ),(0.50 ± 0.23), (0,46 ± 0.27)mg/L, respectively; the 24.0 h excreting rates of fluoride and aluminum from urine were 52.90%(4.64/8.77) and 12.38% (1.37/11.07), respectively. The fluoride concentrations in serum samples collected before and at 0.5,1.0,2.0,3.0,24.0 h after drinking brick tea solution were (0.06 ± 0.02), (0.14 ± 0.01 ), (0.21 ±0.04), (0.17 ± 0.04), (0.10 ± 0.03), (0.04 ± 0.01 )mg/L, respectively, and aluminum in those were (0.30 ± 0.05),(0.27 ± 0.01 ), (0.30 ± 0.12), (0.34 ± 0.19), (0.30 ± 0.10), (0.27 ± 0.09)mg/L, respectively. Conclusions Fluoride in brick tea is easyly to be absorbed and excreted through kidney, and the kidney excretory capacity of aluminum in brick tea is limited. Levels of fluoride and aluminum in urine could be taken as the indicators for monitoring the prevalent extent and evaluating preventive experiment's effect on the brick tea fluoride-aluminum toxicity; and fasting serum fluoride and aluminum levels could be taken as indicators for estimating the toxic condition of the brick tea fluoride-aluminum toxicity.

Myocardial perfusion evaluation post percutaneous transluminal septal myocardial ablation in patients with hypertrophic obstructive cardiomyopathy by ⁹⁹Tc(m) MIBI SPECT MPI / 中华心血管病杂志

<p><b>OBJECTIVE</b>To evaluate the myocardial perfusion and function in patients with hypertrophic obstructive cardiomyopathy (HOCM) before and after percutaneous transluminal septal myocardial ablation (PTSMA).</p><p><b>METHODS</b>Sixty-eight patients with hypertrophic obstructive cardiomyopathy were included and (99)Tc(m)-MIBI SPECT MPI was applied before and at 1 week after PTSMA, six-month follow-up was finished in 11 patients. Semi quantity and QGS quantity perfusion and function assessment was performed in 17 LV segments.</p><p><b>RESULTS</b>Myocardial perfusion post-PTSMA was significantly reduced in 98% patients, especially in basal anterosepta, basal interseptal, mid-anteroseptal, mid-interseptal and apical septal segments compared with pre-PTSMA (all P < 0.05). Perfusion was significantly increased at 6 months follow-up than at 1 week post-PTSMA but still lower than pre-PTSMA (all P < 0.05). LVEF (evaluated by gated SPECT) was similar before and after the procedure (P > 0.05). Regional wall motion after PTSMA was lower than pre-PTSMA in basal anterior, basal anteroseptal, basal interseptal and basal inferior (P < 0.05). Regional wall thinkening was lower than pre-PTSMA in basal interseptal, mid-anteroseptal, mid-interseptal (P < 0.05).</p><p><b>CONCLUSIONS</b>(99)Tc(m) MIBI SPECT can be used to monitor myocardial perfusion post PTSMA in patients with HOCM.</p>

Novel frame-shift mutation of 540 A deletion in GP IIb gene from a patient with Glanzmann thrombasthenia / 中南大学学报(医学版)

OBJECTIVE@#To explore the molecular mechanism of Glanzmann thrombasthenia (GT).@*METHODS@#All 45 exons of alphaIIb and beta3 subunit genes as well as their splicing sites were amplified by polymerase chain reaction(PCR) with 40 primer pairs, and then the PCR products were used to screen the gene mutation by single strand conformation polymorphism-polyacrylamide gel electrophoresis (SSCP-PAGE). The mutation was further confirmed by direct DNA sequencing.@*RESULTS@#A DNA band alterated migration was detected after SSCP-PAGE. DNA sequencing showed that a base deletion within the band at the site of 540 in GPIIb gene(540A) was found.@*CONCLUSION@#The frame-shift mutation caused by the deletion of 540A in GPIIb gene is a novel mutation which is a genetic defect in patients with GT.

3 polymorphisms of gene GP IIb and the function of GP IIb T13959 G in the platelet transfusion refractoriness / 中南大学学报(医学版)

OBJECTIVE@#To investigate the relationship among 3 polymorphisms of GP IIb and the function of GP IIb T13959 G in the platelet transfusion refractoriness(PTR).@*METHODS@#The 26th exon, the 30th exon and the 21st intron of gene GP IIb in 110 individuals were amplified by polymerase chain reaction (PCR), and the PCR products were analyzed with single-strand conformation polymorphism(SSCP) and sequenced to investigate whether there was linkage among the polymorphisms of the gene. Human platelet antigen-3 (HPA-3) gene frequency was detected by Fok I enzyme in 147 patients with hematologic diseases, and was compared with that in 110 normal individuals. Forty-four patients who received apheresis platelet transfusion repeatedly were randomly divided into the HPA-3 homotype group and the control group. The antibodies of the platelet were detected after 3 times of platelet transfusion.@*RESULTS@#There were polymorphisms of gene GP IIb in the 26th, 30th exon and the 21st intron, and the mutations were: T changed into G in 13,959 th of the 26th exon; C changed into T in 16,997 th of the 30th exon; the 9 bps deletion occurred in 11,996-12,004 th of the 21st intron. The 3 polymorphisms had synchronization in the individuals. The results of Fok I enzyme indicated that the frequency of HPA-3a was 83.6% (92/110)and 81.9%(119/147), and that of HPA-3b was 16.4%(18/110) and 19.1%(28/147) in the normal individuals and the patients respectively. There was no significant difference between the patients and normal individuals (P>0.05). After the platelet transfusion, the antibodies of all the cases of the homotype platelet transfusion were negative, but the antibodies in 2 cases of the control group were positive, and there was antibody to HPA-3a in one of the antibodies positive cases.@*CONCLUSION@#(1)There is close linkage among the polymorphisms of gene GP IIb, which is T->G in 13 959 th of the 26th exon, C->T in 16,997 th of the 30th exon, and the 9 bps deletion in 11,996-12,004 th in the 21st intron. (2)The gene frequency of HPA-3a/3b is similar in the normal individuals and patients with hematologic diseases. (3) HPA-3 system may be one of the reasons for PTR in Chinese.

Expression of coagulation factor IX in human CD34 + hematopoietic stem cells by adeno-associated virus 2 / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the expression of human coagulation factor IX (hFIX) gene in human umbilical cord blood (CB) CD34+ cells which was transfected with recombinant adeno-associated virus 2 (rAAV-2).</p><p><b>METHOD</b>The CD34+ cells were transfected with rAAV-2/hFIX and cultured for 21 days for inducing differentiation into myeloid, erythroid and megakaryocytes, respectively. The expression of hFIX was studied at mRNA, protein and biological activity levels. The cytotoxicity of rAAV-2 to CD34+ cells was evaluated by cell proliferation, cell vitality, CD antigen expressions and CFU yields.</p><p><b>RESULTS</b>The hFIX mRNA was expressed in the cultured cells which was verified by RT-PCR and DNA sequencing. An elevated level of hFIX expression and biological activities were detected with a maximum amount of 14.10 ng/10(6) cell x 24 h. During the period of 21 day culture, the cell vitality, cell proliferation, CD antigen expression and CFU yields between the transfected and un-transfected groups had no difference(P > 0.05).</p><p><b>CONCLUSION</b>The human CB CD34+ cells are able to produce functional hFIX after transduction by rAAV-2/hFIX. The cell proliferation and differentiation capacities of the host CD34+ cells were not affected by rAAV-2.</p>

Effect of GPI-PLD on adhesion function of bone marrow mononuclear cell from patients with myeloid leukemia and its mechanism / 中国实验血液学杂志

To explore the effect of glycosyl-phosphatidyl inositol-specific phospholipase D (GPI-PLD) on the adhesion function of bone marrow mononuclear cell from patients with myeloid leukemia and analyze its mechanism, the activity of GPI-PLD in bone marrow mononuclear cell from the patients were measured by using GPI-anchored placental alkaline phosphatase (PLAP) as substrate and Triton-X114 partitioning; the adhesion rate and CD24 expression of these cells were measured by MTT and immunohistochemical method respectively, when these cells were or were not treated by 1 mmol/L 1,10-phenanthroline for 5 hours. The results showed that the GPI-PLD activity of bone marrow mononuclear cells from the patients was significantly inhibited after being treated by 1 mmol/L 1, 10-phenanthroline for 5 hours [(42.08 +/- 7.21)% vs (5.4 +/- 2.96)%], while the adhesion rate and the expression of CD24 of these cells were increased [(49.78 +/- 26.73)% vs (61.19 +/- 29.14)%, (16.02 +/- 9.68)% vs (18.5 +/- 11.14)%, respectively)]. It is concluded that depression of GPI-PLD activity can increase the adhesion rate of bone marrow mononuclear cells from the patients while the CD24 expression is enhanced.

Assessment of diastolic function in patients with hypertrophic cardiomyopathy by Doppler tissue imaging / 中国医学科学杂志(英文版)

<p><b>OBJECTIVE</b>To determine the clinical application of pulsed Doppler tissue imaging in assessing the left ventricular diastolic function and in discriminating between normal subjects and patients with hypertrophic cardiomyopathy with various stages of diastolic dysfunction.</p><p><b>METHODS</b>We measured the peak diastolic velocities of mitral annulus in 81 patients with hypertrophic cardiomyopathy with various stages of diastolic dysfunction and 50 normal volunteers by Doppler tissue imaging using the apical window at 2-chamber and long apical views, respectively. The myocardial velocities were determined with use of variance F statistical analysis.</p><p><b>RESULTS</b>Early diastolic myocardial velocities of mitral annulus were higher in normal subjects than in patients with hypertrophic cardiomyopathy with either delayed relaxation, pseudonormal filling, or restrictive filling. However, peak myocardial velocities of mitral annulus during atrial contraction were similar in normal subjects and patients with hypertrophic cardiomyopathy.</p><p><b>CONCLUSION</b>Doppler tissue imaging can directly reflect upon left diastolic ventricular function. Early phase of diastole was the best discriminator between control subjects and patients with hypertrophic cardiomyopathy.</p>

Effects of integrin alpha IIb(R995A) mutation on receptor affinity and pp125 (FAK) phosphorylation / 中国医学科学杂志(英文版)

<p><b>OBJECTIVE</b>To investigate the role of cytoplasmic domain of integrin alpha IIb in platelet signal transduction.</p><p><b>METHODS</b>Binding capacity of integrin alpha IIb(R995A) to antibody platelet activation complex-1 (PAC-1) and pp125 focal adhesion kinase (FAK) phosphorylation of cells were detected by flow cytometry, immune precipitation, and Western blotting.</p><p><b>RESULTS</b>Without activation, wild-type alpha IIb beta3 Chinese hamster ovary (CHO) cells failed to bind to PAC-1, but mutant chimera alpha IIb(R995A)beta3 CHO cells were able to bind with PAC-1. Furthermore, phosphorylation of pp125 (FAK) in wild-type alpha IIb beta3 CHO cells occured only when cells were adhered to fibrinogen, but could not be detected in bovine serum albumin suspension. However in the mutant chimera group, it could be detected in both conditions.</p><p><b>CONCLUSION</b>The mutation in integrin alpha IIb(R995A) alters its affinity state as a receptor, thus also mediating cytoplasmic signal transduction leading to the phosphorylation of pp125 (FAK) without ligand binding.</p>