Objective To investigate the feasibility of preparing HCV-1 diagnostic microarray probes using the technique of cDNA fragments library construction. Methods The full-length cDNAs of HCV of subtypes 1a and 1b were digested with restriction endonuclease Sau3A I, and the resulted fragments were cloned into the pMD18-T vector. Positive clones were isolated and identified by sequencing. Results A total of 57 different fragments were obtained, and sequence analysis showed that all the fragments ranging from 200 to 1000bp were specific gene fragments of HCV genotype 1, which can be efficiently used as probes in microarray prepapration. Conclusion The method of preparing microarray probes by construction of cDNA fragments library was effective, quick and simple.

Objective To prepare the microarrays for joint detection of HBV and HDV. Methods The specific primers of PCR were designed with Primer Premier 5.0 program according to the conserved regions of HBV and HDV. The PCR fragments were purified and cloned into the pMD18-T vectors. The recombinant plasmids were extracted from positive clones and the target gene fragments were sequenced. The DNA microarray was obtained by spotting PCR products onto the surface of glass slides by robotics. Restriction display PCR (RD-PCR) was used to label the samples. Results After the sequences were aligned, we found that the products of PCR amplification were the specific gene fragments of HBV and HDV. The hybridized signals on gene chips indicated that the specificity and sensitivity of DNA microarray for joint detecting the HBV and HDV were satisfactory. Conclusion Using PCR amplification products to construct gene chips is a quick, simple and effective method for clinical diagnosis of HBV and HDV. Further application of restriction display PCR technique in labeling the sample may expedite and raise the sensitivity in multi-virus detection by microarray technology.

Objective To investigate the applieation of restriction display (RD) technique in the preparation of HCV probes of clinical genotyping microarray. Methods Restriction enzyme Sau3A Ⅰwas chosen to digest the full-length HCV cDNAs of three distinct subtypes, i.e.1a, 1b and 2a. The resultant restrictive fragments were then ligated with universal adapters. PCR primers were designed to match the universal adapters but with one "nesting" base overhanging at the 3′- end. The PCR reactions were performed by ten pairs of different primer combinations. The differential genes were separated through polyacrylamide gel electrophoresis and silver staining. The second-round PCR was performed using the isolated bands as PCR templates. The purified PCR products were then cloned into T-vectors. The recombinant plasmids were extracted from positive recombinant clones and the target gene fragments were sequenced. Results The target HCV gene fragments ranging from 200 to 900bp were isolated and sequenced, which were correlated precisely with the RFLP (Restriction Fragment Length Polymorphism) prediction. A total of 66 different fragments were obtained, averaging about 22 for each subtypes. These fragments could be further used as probes in HCV microarray preparations. Conclusion RD technique is of great value in obtaining a large number of equal sized gene probes, which provide a swift protocol in generating DNA probes for the preparation of microarrays.

Objective To investigate haemodynamic change during hemodialysis and analyze effects of cardiac index (CI) on hemodynamic parameters and associated influencing factors in maintenance hemodialysis (MHD) patients.Methods Seventy-five patients bearing an arteriovenous fistula (AVF) entered the study.Cardiac output (CO),cardiac index (CI),central blood volume (CBV) and peripheral vascular resistance (PR) were determined by ultrasound dilution technique at the end of 1 hour,2 hours and 3 hours of dialysis.AVF blood flow (Qa) was also measured with the same device before haemodynamic parameters investigation.Results Mean age of patients was (55.84 ±12.39) years old (range 21-81 years) and 43 patients (57.3％) were female.Systolic arterial pressure,SV,CO,CI and CBV were significantly declined and no significant change for diastolic arterial pressure and heart rates at the end of 2 hour and 3 hour hemodialysis,whereas PR was increased gradually during hemodialysis.Patients were divided into there groups with CI less than 2.5 L·min 1·(m2)-1,2.5-4.2 L·min-1·(m2)-1 and more than 4.2 L·min-1 ·(m2)-1 at the end of 1 hour dialysis.Statistically significant decreasing SV,CO and increasing PR were detected in patients with CI＜2.5 L·min-1· (m2)-1 and 2.5-4.2 L· min-1· (m2)-1,compared with CI＞4.2 L·min-1· (m2)-1 group (P＜0.01).The hemodynamic change was the most obvious in the group of CI less than 2.5 L· min-1· (m2) 1,and no significant changes happened in CI＞4.2 L·min-1 ·(m2)-1 group.Some factors were found to be associated to CI values.Qa and systolic arterial pressure had positive relationship with CI,while age and diabetes had negative relationship with CI.Conclusions Systolic arterial pressure,CO,CI and CBV decrease and PR increases during hemodialysis.Obvious change occurs when CI is less than 2.5 L·min-1· (m2)-1.CI is associated with Qa,systolic arterial pressure,age and diabetes.

Objective To analysis the clinical and pathological features, results of laboratory tests and prognosis of nephrotic syndrome (NS) in patients with non-heamatological maligancy. Methods The data were collected from 25 patients who presented with NS around the diagnosis of non-heamatological malignancy. Results Twenty-five cases were investigated (age: (56.6±17.7) years; male/female ratio: 20/5). Malignancy and NS occurred within one year in 92% patients. There was a wide distribution of malignancy with involvement of 36% in digestive system and 20% in respiratory system. Ten patients (40%) presented with NS as their initial manifestation. Heamaturia appeared in 67% patients and acute renal insufficiency was complicated in 12% cases before treatments. Some other non-specific laboratory tests were found including elevated serum gamma-globulin in 50% and anemia not related with renal failure in 28% cases. Membranous nephropathy was the most common pathological changes in 67% cases. Although NS still continued for several weeks in 8 of 9 cases after surgery and/or chemical therapy, glucocorticoids was helpful to achieve the remission in these patients. However, no remission was achieved in patients without the treatment for malignancy. Conclusion Malignancy may present with NS as its initial manifestation. It should be inspected routinely and regularly in elder patients with NS, especially in those with membranous nephropathy, as well as gamma-globulinemia and anemia.

Objectives:To preparing the probes of cDNA microarray in detection of the Hepatitis D virus. Methods:The specific primers of PCR were designed accordion to the conserved region of HDV. Results: Sequences were aligned , and the results showed that the products of PCR amplification were the specific gene fragments of HDV. Conclusions:Using PCR amplification products to prepare gene chip probe was a quickly, simple effective method.

Objective To investigate the involvement of mitogen-activated protein kinase(MAPK)and PI3K(phosphatidylinositol 3 kinase)/ Protein kinase B(Akt) signal transduction pathways in the mechanism of myocardium remodeling in patients with congestive heart failure(CHF).Methods Thirty nine patients of mitral valve disease with CHF were randomly selected and 30 cases of healthy persons were included as controls.Cardiac function parameters were measured by echocardiography.Concentration of AngⅡ in plasma and myocardial tissues was determined by radio immunoassay.Activity of PKC was determined by using competive prote in binding method,activity of MAPK was detected by the methods of immunoprecitipation.Immunoprecitipation was used to assay the protein expression and phosphorylation of PI3K and Akt(Protein kinase B),protein expression of C-FOS and ?-skeletal-actin in myocardial tissues.Results Pathological changes of myocardial tissues in CHF with valvular heart diseaseshowed typical myocardial remodeling.The hypertrophy was dominant at early stagy of CHF,while at end stage the characteristics include disordered alignament of the myocytes,the discontinuity and dissolving of cardiomyofibrills,destroyed subcellular organs,and the hyperplasia of interstitial tissue.AngⅡ concentration in plasm and myocardial tissues in patients with CHF was higher than those in the control group(P

Objective To investigate the clinical characteristics of hypertrophic cardiomyopathy (HCM) in the elderly. Methods The clinical characters of HCM in 70 elderly patients were retrospectively analysed. Results Among the 196 identified HCM patients, 70 were elders. Out of them, 7 patients(10.0%) were suspected as HCM according to the clinical symptoms, 29 patients (41.4%)were suspected as other cardiac diseases, 34 (48.3%) were diagnosed HCM due to other reasons. Among the 70 patients, 12 patients(17.1%) had history of cerebrovascular diseases, 54 (77.1%) manifested symptoms after 45 years of age and 4 (5.7%) showed no obvious symptoms. Among them, 18 patients were examined by UCG for two times and HCM was diagnosed at the second time. Conclusions HCM in the elderly is not an uncommon disease. The onset of cardiac symptoms is relatively late in the elderly HCM and frequently misdiagnosed; cerebrovascular diseases are commonly seen in elderly HCM patients.

Objective:To understand the state of Chlamydia pneumoniae (Cpn) infection in patients with coronary hear disease (CHD), and explore the relationship between Cpn infection and the gonesis and progressin of CHD.Methods:By means of PCR and ELISA, Cpn IgG antibody and nucleic acid were detected in 159 patients with CHD and 41 control subjects.Results:The positivity rate of Cpn DNA was 43 40%(69/159) in the patient group and 7 32%(3/41) in the control group, showing obvious difference between the two groups( P

Objective To observe the cerebral hemodynamics in ischemic stroke patients after compound anisodine acupoint injection.Methods 40 stroke patients were measured with transcranial color Doppler before and 4 h,24 h after compound anisodine acupoint injection.Results After injection,the velocity of the cerebral artery showed some increase,the end diastolic velocity of the ophthalmic artery increased and the pulsitility index decreased(P<0.05).Conclusion The compound anisodine acupoint injection may improve the flow velocity of ophthalmic artery and cerebral artery.