Objective To evaluate the accuracy and clinical practicality of DNA chip in comparison with serology in typing of human leukocyte antigen A (HLA-A) in Han's individuals of donor-recipients of transplantation. Methods 120 peripheral blood samples were obtained from donor-recipients of transplantation. Each sample was divided into two parts and HLA-A antigens were identified by DNA chip in one part and by serology in another. Samples in which the HLA-A typing results by these 2 methods were discordant were verified by polymerase chain reaction with sequence specific primers (PCR-SSP). Accuracy and clinical practicability of both methods were compared according to the typing results. Results Serological typing for HLA-A took 3 h, while DNA chip typing 4. 5-5 h. 112 samples have been typed successfully. Typing results were same in 91 samples and discordant in 21 cases. The verified results showed that DNA chip made 2 incorrect typing and the error rate was 2%. Meanwhile, serology made 19 mistakes, consisting of 5 antigens being incorrectly interpreted and 14 "blanks" turning out to be definable alleles. The discrepancy rate was 17 %. Conclusions DNA chip typing for HLA-A is suitable for clinical application in Chinese Han's population with a greater precision than serology. It may replace the serology in future after being improved and perfected.

Objective To explore the instructive value of preoperative splenic artery CTA examination on Laparoscopic splenectomy (LS). Methods From January 2008 to February 2010,36 cases requiring Laparoscopic splenectomy (LS) were selected randomly and divided into two groups: CTA group and non-CTA group, 18 cases in each group. As for the CTA group,splenic artery CTA examination was performed before operation,to understand splenic artery and its branches anatomy type and track and then individualized surgical treatment was developed. As for the non-CTA group,a routine surgical procedure was performed. The indices before and during the operation were recorded and compared through χ2 test The operating time and the amount of bleeding in the two groups were compared using t-test Results There were no significant difference between the indices recorded by splenic artery CTA examination and those by laparoscopic splenectomy intraoperation ( P > 0. 05 ) . The operating time was (124. 32 ±21.43) mins in the CTA group, which was significantly shorter than that in the non-CTA group ((148.27 ±28. 36)mins) (P <0. 05). The amount of blood in the CTA group was significantly less than that in the non-CTA group( ( 80. 50 ± 16. 42) ml vs. (101. 35 ± 26.25 ) ml). Conclusions Splenic artery CTA can identify splenic artery and its branches anatomy type and track before LS,therefore guide the development of individualized surgical treatment It increases the safety of LS, reduces bleeding and shortens the operational time. It has an instrutive value to laparoscopic splenectomy.

Objective To study the correlation between amino terminal B-type natriuretic peptide precursor(NT-proBNP) and risk factors of acute coronary syndrome (ACS) with elder patients.Methods One hundred and twenty-eight ACS patients were divided into unstable angina pectoris (UAP) group with 52 samples,ST elevation myocardial infarction (STEMI) group with 35 samples and non-ST elevation myocardial infarction (NSTEMI) group with 41 samples.Meanwhile 45 healthy elder people were adopted as control group.Firstly,the subjects of blood pressure,body mass index (BMI) and smoking numbers were measured.Secondly,venous blood was collected to assay NT-proBNP,cardiac troponin Ⅰ (cTn Ⅰ),homocysteine (Hcy),blood-lipoids and C-reactive protein(CRP).Lastly,ultrasonic cardiogram was used to test left ventricular end diastolic diameter(LVEDD),left ventricular end systolic diameter (LVESD) and left ventricular ejection fraction (LVEF).In addition,correlation analysis was researched between NT-proBNP and other factors.Results NT-proBNP levels of UAP,STEMI and NSTEMI groups were significantly higher than the control group ((794.18±182.64) ng/L,(872.43±245.67) ng/L,(557.25±163.81) ng/L) and (125.84±59.27) ng/L,P ＜ 0.05).NT-proBNP was positive correlation with systolic blood pressure,diastolic blood pressure,Hcy and CRP (r=0.182,0.176,0.281,0.191;P=0.040,0.043,0.001,0.031),however negative with LVEF(r=-0.247,P =0.005).Conclusion NT-proBNP level is sensitive to monitor ACS variety,and it is significant to test NT-proBNP combining Hcy,CRP,and cTn Ⅰ for diagnosing and treating ACS.

Objective To study the anti-injury effects of grape seed proanthocyanidin (GSP) on the umbilical vein endothelial cell (HUVEC)injured by hydrogen peroxide(H2O2) and its mechanism. Method Oxidative damage model of endothelial cell was induced by H2O2,and the cells were divided into five groups,including normal groups,oxidative damage group and three (100,50,10 ?g/ml) GSP pretreated groups. MTT assay was used to detect the protection of GSP,Reverse transcription PCR (RT-PCR) was used to detect the protein expression in mRNA level of monocyte chemoattractant protein (MCP-1) and flow cytometry(FCM) was used to detect apoptosis rate and quantitate the expression of intercellular adhesion molecule(ICAM-1)and vascular cell adhesion molecule ( VCAM-1). Results The cell optical density in oxidation damaged group was lower than that in normal control. When pretreated by GSP,the OD value was increased,and was insignificantly different between high GSP group and normal control. In H2O2 damage group,the expression of MCP-1 mRNA and the apoptosis rate were increased,and the numbers of positive cells which express ICAM-1 or VCAM-1 were obviously more than those in pretreated groups respectively.Futhermore,these effects were dose dependent. Conclusion GSP has the protective effect on oxidative damage of HUVEC. The mechanism may be related to stabilization of cell wall and inhibition of inflammatory reaction and apoptosis induced by ICAM-1,VCAM-1 and MCP-1.

Objective To study the heart rate variability and arrhythmia characteristics in the elderly patients with unstable angina and negative psychology and to observe the changes before and after treatment. Methods Eighty patients aged ≥ 65 years with unstable angina were recruited in the study, and divided into two groups according to the determination of Hospital Anxiety and Depression Scale: 36 patients in the negative psychology group( NP group) and 44 in the non-negative psychology group ( non-NP group). All patients were monitored by Holter. Results The SDNN, SDANN, SDNNidex, rMSSD and PNN50 in the NP group were (86.8 ±14.1 ) ms, (78.1 ± 11.2 ) ms, (22.9 ± 4.5 ) ms, ( 6.3 ± 1.2 ) % and (40.3 ± 16.4) ms, respectively, which were significantly decreased compared to that in the non-NP group [ ( 138.9 ± 37.1 ) ms, ( 123.3 ± 32.5 ) ms, (25.7 ±12.3)ms,(9.2 ±5.3)% and (47.8 ± 10.7)ms,respectively] (t =4.20,2.52,2.32,2.28 and 2.20,Ps ＜0.05)The prevalence of ventricular arrhythmia, sinus tachycardia, ischemic ST-T changes and heart rate variability (HRV) (55.56%, 13.89%, 33. 33% and 52. 78%, respectively) significantly increased in the NP group,compared to those in the non-NP group(9.09% ,2.27% ,6.82%, 18.18% ,respectively) ( χ2 = 16.89,4.18,4.33 and 4.99, respectively, Ps ＜ 0.05 ). After treatment with Prozac ( fluoxetine hydrochloride scientific name), SDNN,SDANN,rMSSD,PNN50,SDNNindex significantly increased from (84.7 ± 34.0) ms, (79.2 ± 39.6) ms, (23.4 ±7.5)ms,(5.1 ±2.1)% and (38.1 ± 11.8)ms to (102. 1 ±29.6)ms,(94.2 ±26.7)ms,(35.6 ± 10.2)ms,(9.9 ± 7.5 ) % and (63.3 ± 13.8 ) ms ( t = 3.60,2.43,2.34,2.26 and 2.30, P ＜ 0.05 ). After treatment,ventricular arrhythmia,sinus tachycardia,ischemic ST-T changes and HRV significantly decreased compared to that before treatment in the NP group (χ2 = 5.58,4.33,4.24 and 4.06 ,P ＜ 0. 05 ). Conclusions In patients of coronary heart disease with unstable angina and negative psychologically , HRV decreased , and they are prone to ventricular arrhythmias,sinus tachycardia and ischemic ST-T changes. Treatment with Prozac could decrease arrhythmia and improve cardiac ischemia arrhythmias, sinus tachycardia and ischemic ST-T changes. Treatment with Prozac could decrease arrhythmia and improve cardiac ischemia.

Objective To investigate the correlation between serum cystatin-C (Cys-C) levels and cognitive impairment.Methods A perspective study involving 273 patients in our hospital was performed from 2013 to 2015.They were divided into the cognitive impairment group and the control group.Cys-C measurement results and cognitive impairment assessment scores were collected.Possible influence factors were adjusted,and the correlation between Cys-C levels and mild cognitive impairment was analyzed.Results The distribution of age,hypertension,diabetes mellitus,smoking,drinking,dyslipidemia,and creatinine showed significant differences between the groups at different Cys-C levels (all P＜0.05).There was a significant difference in the Cys-C level between the cognitive impairment group and the control group [(0.727±0.082) mg/L vs.(0.514±0.045)mg/L,t=23.68,P＜0.01].The Cys-C level was negatively correlated with the scores of mini-mental state examination (MMSE) and Montreal cognitive assessment (MOCA) in the cognitive impairment group (r =-0.318 and -0.572,P＜0.05).The incidence of cognitive impairment was elevated with increasing Cys-C levels (x2 =13.12,P＜0.01).Logistic regression analysis showed that high levels of Cys-C (OR=3.298,95％ CI:1.417-7.675,P=0.001),history of diabetes mellitus (OR=7.971,95％ CI:3.036-31.562,P =0.03),education level (OR =2.237,95 ％ CI:1.022-4.896,),smoking (OR =5.692,95 ％ CI:1.060-2.614),drinking (OR=1.227,95％ CI:0.083-1.228),and dyslipidemia (OR=2.267,95％ CI:1.177-4.366) are independent risk factors for cognitive impairment.Conclusions High serum cystatin C levels are closely correlated with the risk of cognitive impairment occurrence.

Aim To make ethanol from bistortae(EFB)from traditional Chinese medicine bistortae,and investigate the protection on the injury of the human umbiliar vein endothelial cell(ECV304)induced by hydrogen peroxide(H2O2).Methods ECV304 cells were cultured and divided by five experimental groups including normal,injury model induced by H2O2 and three pretreated groups with EFB;flow cytometry(FCM)methods Annexin V/PI staining and PI staining were used to analyze the effect of the medicine on the apoptosis and cell cycle;reverse transcription PCR(RT-PCR)was used to detect the mRNA expression level of apoptosis associated protein caspase-3 in all experiment-al groups.Results FCM Annexin V/PI staining result showed early and advanced stage apoptosis rate was lower when predisposed than that of the damaged group,FCM PI staining showed there were more cells in G0/G1 stage in oxidation damage group,but less in S stage,and hypodiploid peak could be observed,RT-PCR showed that the expression of caspase-3 was weakened when predisposed by EFB than without this disposal(P

ObjectiveTo explore the effects of hepatitis B virus X protein (HBx) on the expression and promoter methylation of the p16 tumor suppressor gene, and to investigate the epigenetic role of HBx in the development and progression of hepatitis B virus (HBV)-associated hepatocellular carcinomas (HCC). MethodsExperiments were performed in the human hepatoblastoma cell line HepG2, HepG2 cells expressing green fluorescent protein (HepG2/GFP), and HepG2 cells stably expressing GFP-HBx fusion protein (HepG2/GFP-HBx). Western blot was used to determine the expression levels of the p16 protein in HepG2 cells, HepG2/GFP cells, and HepG2/GFP-HBx cells. HepG2/GFP-HBx cells were treated with a universal inhibitor of DNA methyltransferase (DNMT), 5-aza-2'-deoxycytidine (5-aza-2'-dC). Methylation-specific polymerase chain reaction (MSP) was used to determine the promoter methylation of the p16 tumor suppressor gene in HepG2 cells, HepG2/GFP cells, and HepG2/GFP-HBx cells treated with or without 5-aza-2′-dC. Multiple-group comparison was made by analysis of variance. ResultsAccording to the results of Western blot, HepG2/GFP-HBx cells had a significantly lower expression level of the p16 protein than HepG2 cells and HepG2/GFP cells (P=0.0007; P=00014); there was no significant difference in the expression level of the p16 protein between HepG2/GFP and HepG2 cells (P＞0.05). The MSP assay revealed partial CpG methylation in the p16 promoter region in HepG2/GFP-HBx cells. No promoter methylation was detected in HepG2 cells or HepG2/GFP cells. Non-methylation in the p16 promoter region was restored in HepG2/GFP-HBx cells treated with 5-aza-2′-dC. ConclusionIn the hepatoblastoma cell line, HBx down-regulates the expression of the p16 tumor suppressor gene by inducing methylation in its promoter region. The DNMT inhibitor, 5-aza-2′-dC, restores non-methylation in the p16 promoter region. The reversible modification provides new insights for the treatment and prevention of HBV-associated HCC.

OBJECTIVETo compare two different routes of totally implantable venous access ports (TIVPs) from the upper arm vein and the subclavian vein in terms of complications for patients with gastrointestinal malignancy.

METHODSPatients who underwent implantations of TIVPs from September 2013 to January 2015 were retrospectively evaluated. The outcome measurements were rates and types of postprocedural early-stage and long-term complications.

RESULTSA total of 208 patients(upper arm vein group, 86; subclavian vein group, 122) were included in this study. All TIVPs were implanted successfully. The rate of catheter displacement was higher in upper arm vein group(14.0% vs 5.7%, P=0.04), while other postprocedural early-stage complications had no significant difference between the two groups. The occurrence of transfusion obstacle and rates of overall postprocedural long-term complications were significantly lower in upper arm vein group than that in subclavian vein group(1.2% vs. 9.8%, P=0.02; 7.0% vs. 27.0%, P=0.01, respectively).

CONCLUSIONCompared with subclavian vein group, upper arm vein group has lower postprocedural long-term complication rates and is recommended as a safe and comfortable choice for port implantation.

Objective: To observe ascitic interleukin-7 expression level in cirrhotic patients complicated with spontaneous bacterial peritonitis, and to detect the effect of recombinant human IL-7 on CD4⁺ and CD8⁺T lymphocyte function. Methods: A total of 84 patients with liver cirrhosis who were hospitalized from August 2017 to April 2018 were selected. Among them, 51 cases were complicated with cirrhosis and untainted ascites, and 33 cases were cirrhosis complicated with spontaneous bacterial peritonitis. Peripheral blood and ascites were collected routinely. The levels of IL-7 in peripheral blood and ascites were measured by enzyme-linked immunosorbent assay. CD4⁺T cells and CD8⁺T cells were purified from ascites, and were stimulated with recombinant IL-7. Cellular proliferation, key transcription factors for mRNA, and cytokines production by CD4⁺T cells in response to IL-7 stimulation was measured. mRNA expression corresponding to perforin, granzyme B, and granulysin as well as cytokines production by CD8⁺T cells was also measured in response to IL-7 stimulation. Cytolytic and non-cytolytic activity of CD8⁺T cells in response to IL-7 stimulation was also investigated in both direct and indirect contact co-culture system. Measurement data of the normal distribution were compared between the two groups by Student’s t-test and the data before and after stimulation were compared by paired t-test. Measurements that did not conform to normal distribution were compared between the two groups using Mann-Whitney U test, and data before and after stimulation were compared using Wilcoxon paired test. Results: There was no significant statistical difference in serum IL-7 levels between the two groups [(5 001 ± 1 458) pg/ml vs. (4 768 ± 1 128) pg/ml, P = 0.41]. The level of ascitic IL-7 in cirrhotic patients complicated with SBP was significantly lower than cirrhosis patients with untainted ascites [(966.4 + 155.8) pg/ml vs. (792.1 + 126.4) pg/ml, P < 0.01]. Recombinant IL-7 stimulation promoted the proliferation of CD4⁺ and CD8⁺T cells from ascites in patients with liver cirrhosis complicated by SBP. T-bet mRNA relative expression and IFN-γ secretion in CD4⁺T cells was also elevated in response to IL-7 stimulation in vitro. Moreover, IL-7 stimulation also increased the mRNA expressions of perforin, granzyme B, and granulysin as well as productions of IFN-γ and TNF-α by CD8⁺T cells. Recombinant IL-7 stimulation elevated cytolytic and non-cytolytic activity of CD8⁺T cells from ascites in patients with liver cirrohosis complicated by SBP, which manifested as increased target cell death and IFN-γ production in both direct and indirect contact co-culture system. Conclusion Ascitic IL-7 promotes T lymphocyte function in patients with liver cirrhosis complicated with SBP.