OBJECTIVETo develop a method for detection of coxsackie B virus type 1-6 by RT-PCR.

METHODSA pair of primers were designed to amplify all types of coxsackie B virus 1-6 efficiently. The PCR product was hybridized in micro-wells in which 6 type specific oligonucleotide probes had been coated respectively, colorimetric detection was performed to discriminate the types of coxsackie B virus.

RESULTSThis method was shown to be concordant with the IgM ELISA, 71.7% of anti-coxsackie B positive cases could be detected by RT-PCR.

CONCLUSIONThe RT-PCR method can type coxsackie B virus efficiently and provides a tool for clinical diagnosis and epidemiological investigation.

DNA Primers ; Enterovirus B, Human ; classification ; genetics ; isolation & purification ; Enterovirus Infections ; diagnosis ; virology ; Enzyme-Linked Immunosorbent Assay ; Humans ; Immunoglobulin M ; blood ; Reverse Transcriptase Polymerase Chain Reaction ; methods

OBJECTIVEBrugada syndrome is an inherited channelopathy that characterized by ST-segment elevation in the right precordial lead (V(1)-V(3)) on the electrocardiogram with or without right bundle branch block and related with high risk of sudden cardiac death and structurally normal hearts. The first and only gene linked to this disease is SCN5A, a gene encodes for alpha subunit of the cardiac sodium channel. The objective of this study is to explore SCN5A gene mutations in Chinese patients with Brugada syndrome.

METHODSFour patients diagnosed as Brugada syndrome and nine patients with suspected Brugada syndrome were chosen for the study. The exons in the functional regions of SCN5A gene were amplified with polymerase chain reaction and the amplified products were sequenced with Sanger method. If a mutation was identified, patient's family members were also screened.

RESULTSTwo heterozygous mutations were found in one family diagnosed as Brugada syndrome. One missense mutation was a G-->A transition in the first nucleotide of codon 95 in SCN5A gene exon 3, which was predicted to result in substitution of Valine with Isoleucine (V95I). The other missense mutation was a C-->T transition in the second nucleotide of codon 1649 in SCN5A gene exon 28, which was predicted to result in substitution of Alanine with Valine (A1649V). A heterozygous mutation was identified in one family suspected to have the disease. The mutation was a three nucleotides (TCT) deletion that caused Phenylalanine deletion in codon 1617 in SCN5A gene exon 28. The three mutations were not detected in 100 control chromosomes.

CONCLUSIONSMutation in SCN5A gene is one of the causes of Brugada syndrome in Chinese. Three novel SCN5A gene mutations were identified in Chinese with Brugada syndrome, which expands the spectrum of SCN5A mutations associated with the disease.

Adolescent ; Adult ; Aged ; Brugada Syndrome ; genetics ; Case-Control Studies ; Exons ; genetics ; Humans ; Male ; Middle Aged ; Muscle Proteins ; genetics ; Mutation ; NAV1.5 Voltage-Gated Sodium Channel ; Sodium Channels ; genetics

OBJECTIVETo investigate the effect of preoperative use of viable Bifidobacterium supplement on the intestinal flora, immune status, inflammatory response and prognosis of patients undergoing colorectal cancer resection.

METHODSSixty patients with colorectal cancer were randomized into treatment group (n=30) and control group (n=30). Patients in the treatment group received oral viable Bifidobacterium with routine enteral nutrition and patients in the control group received routine enteral nutrition alone. The intestinal flora of stool was analyzed and stool SIgA, serum IgG, IgM, IgA, IL-6, and C-reactive protein (CRP) were detected.

RESULTSPostoperative Bifidobacterium/E.coli (B/E) ratio in the treatment group decreased significantly as compared to the preoperative ratio (2.01+/-0.36 vs 26.53+/-4.22, P<0.05). However, the ratios remained above one. Both preoperative and postoperative B/E ratios in the control group (0.72+/-0.14, 0.02+/-0.06) were significantly lower than those in the treatment group (P<0.05). Indexes of immunity and inflammation were not significantly different between the two groups (P>0.05). At day 9 after operation, stool SIgA was higher in the treatment group, while serum IgG, IgM, IgA, IL-6, CRP in the treatment group were lower (P<0.05). Postoperative septic complications in the treatment group was less than that in the control group (P<0.05), but other complications and hospital stay were comparable.

CONCLUSIONSIn patients with colorectal cancer, supplementation of viable Bifidobacterium before surgery can improve bacterial dysbiosis and immunity, and can reconstruct the balance of intestinal flora, and reduce infection complication of surgery.

Adult ; Aged ; Aged, 80 and over ; Antibodies ; blood ; Bifidobacterium ; C-Reactive Protein ; analysis ; Colorectal Neoplasms ; immunology ; microbiology ; therapy ; Female ; Humans ; Inflammation ; Interleukin-6 ; blood ; Male ; Middle Aged ; Postoperative Period ; Probiotics ; therapeutic use

AC133 Antigen ; Adult ; Aged ; Antigens, CD ; metabolism ; Carcinoma, Hepatocellular ; diagnosis ; metabolism ; pathology ; Female ; Glycoproteins ; metabolism ; Humans ; Liver Neoplasms ; diagnosis ; metabolism ; pathology ; Male ; Middle Aged ; Peptides ; metabolism ; Prognosis ; Thy-1 Antigens ; metabolism

OBJECTIVETo investigate the role of D-galactose, especially in the structural and functional changes of the immune system in aging.

METHODSSerum levels of advanced glycation end-products (AGE) were determined by ELISA method. Ultra-structures of thymus and spleen were detected by transmission electron microscopy. MTT method was used to determine the lymphocyte proliferation. IL-2 activity was determined by bioassay. Northern blot was used to detect the IL-2 mRNA levels.

RESULTSSerum AGE levels of D-galactose- (P < 0.01) and AGE-treated (P < 0.05) mice (n = 8) were increased significantly. The ultra-structures of thymus and spleen in D-galactose- and AGE-treated mice showed regressive changes similar to those in the aged control group. The lymphocyte mitogenesis and IL-2 activity of spleen were also decreased significantly (P < 0.01, n = 8). The change of IL-2 activity shown by Northern blot resulted from the change of mRNA expression. The AGE plus aminoguanidine group, however, showed no significant change in these parameters in comparison with the young control group (P < 0.01 or P < 0.05, n = 8).

CONCLUSIOND-galactose and AGE lead to a mimic regression change of aging in the immune system in vivo.

Aging ; drug effects ; immunology ; Animals ; Cell Proliferation ; drug effects ; Galactose ; pharmacology ; Glycation End Products, Advanced ; blood ; Interleukin-2 ; metabolism ; Lymphocytes ; drug effects ; immunology ; Mice ; Microscopy, Electron, Transmission ; RNA, Messenger ; metabolism ; Spleen ; drug effects ; immunology ; ultrastructure ; Thymus Gland ; drug effects ; immunology ; ultrastructure

OBJECTIVETo observe the expressions of E-cadherin and beta-catenin in LNCaP and ARCaP cell lines and explore their relationship with the metastasis of human prostate cancer.

METHODSThe expressions and distribution of E-cadherin and beta-catenin in LNCaP and ARCaP cell lines (IF11 and IA8) were detected by Western blot and immunofluorescent staining.

RESULTSThe expression of E-cadherin was high in LNCaP, but absent in IF11 and IA8, while beta-catenin was expressed highly in IF11 and LA8, but lowly in LNCaP. Immunofluorescent staining showed that E-cadherin was mainly in the membrane of LNCaP, while beta-catenin both in the membrane of LNCaP and in the nuclei of IF11 and IA8.

CONCLUSIONE-cadherin and beta-catenin are differently expressed and distributed in prostate cancer cell lines with different characteristics of epithelial-mesenchymal transition (EMT), and the abnormal activation of the beta-catenin signal pathway may be involved in the EMT of prostate cancer cells.

Cadherins ; metabolism ; Cell Line, Tumor ; Humans ; Male ; Prostatic Neoplasms ; metabolism ; pathology ; beta Catenin ; metabolism

BACKGROUNDAlthough endomyocardial biopsy (EMB) plays a crucial role in the final diagnosis in patients with heart failure of unknown etiology, the invasive nature of this technique limits its clinical application in China. The purpose of this study was to evaluate the clinical application of EMB in diagnosing cardiomyopathy with unexplained etiologies in China.

METHODSFifty-three consecutive patients (38 males, age 14 - 67 years, median 43 years) were included in the study who were initially diagnosed as unexplained cardiomyopathy and under EMB biopsy in Peking Union Medical College Hospital from 2006 to 2009. The patients were clinically divided into four groups: dilated, hypertrophic, restrictive and unclassified cardiomyopathy. Biopsies were performed via right internal jugular vein with the use of the bioptome under fluoroscopic guidance. Three to five endomyocardial samples were taken from each patient for light microscopy examination and one sample for electron microscopy was taken if necessary. For each patient, an initial clinical diagnosis, an EMB diagnosis and a final diagnosis prior to discharge were established. All the data were compared and analyzed for the evaluation of clinical utility of EMB in China.

RESULTSIn 26 patients initially diagnosed with restrictive cardiomyopathy (RCM), the etiology of the condition was finally diagnosed using EMB in 15; including 13 amyloidosis and two eosinophilic myocarditis. We employed EMB in 19 patients clinically diagnosed as dilated cardiomyopathy and detected viral myocarditis in one patient, cardiac involvement due to polymyositis in four and doxorubicin-induced cardiomyopathy in one. In five patients with severe left ventricle hypertrophy undergoing EMB, one patient was diagnosed as autophagic vacuolar cardiomyopathy and one as mitochondrial disease. In the remaining three patients with unclassified cardiomyopathy, EMB revealed infiltration of eosinophils as the cause of atrial ventricular block in one patient. Final diagnoses were made in 24 of the total 53 patients (45%) based on the combination of EMB and clinical data. Transient atrial ventricular block in a patient with prior complete left bundle branch block was the only complication occurred during the procedures.

CONCLUSIONThe clinical application of EMB is safe. The combination of EMB and clinical data produced a better understanding of the mechanisms behind the clinically diagnosed cardiomyopathy in China.

Adolescent ; Adult ; Aged ; Biopsy ; methods ; Cardiomyopathies ; classification ; diagnosis ; pathology ; Cardiomyopathy, Dilated ; diagnosis ; pathology ; Cardiomyopathy, Hypertrophic ; diagnosis ; pathology ; Cardiomyopathy, Restrictive ; diagnosis ; pathology ; Female ; Humans ; Male ; Middle Aged ; Myocardium ; pathology ; Young Adult

OBJECTIVETo investigate the inhibiting effects and mechanism of achyranthes bidentata polysaccharide (ABP) and lycium barbarum polysaccharide (LBP) on nonenzyme glycation in D-galactose induced mouse aging model.

METHODSSerum AGE levels were determined by AGE-ELISA, MTT method was used to determine lymphocyte proliferation, IL-2 activity was determined by a bioassay method. Spontaneous motor activity was used to detect mouse's neuromuscular movement, latency of step-through method was used to examine learning and memory abilities of mouse, colormetric assay was used to determine hydroxyproline concentration in mouse skin, pyrogallol autoxidation method was used to determine superoxide dismutase (SOD) activity of erythrocytes.

RESULTSDecreased levels of serum AGE, hydroxyproline concentration in mouse skin and spontaneous motor activity in D-galactose mouse aging model were detected after treated with ABP or LBP, while lymphocyte proliferation and IL-2 activity, learning and memory abilities, SOD activity of erythrocytes, were enhanced.

CONCLUSIONSABP and LBP could inhibit nonenzyme glycation in D-galactose induced mouse aging model in vivo and ABP has a better inhibiting effect than LBP.

Achyranthes ; chemistry ; Aging ; physiology ; Animals ; Disease Models, Animal ; Erythrocytes ; Female ; Galactose ; chemistry ; Learning ; Lycium ; chemistry ; Memory ; Mice ; Mice, Inbred C57BL ; Motor Activity ; Polysaccharides ; pharmacology ; Superoxide Dismutase ; pharmacology

BACKGROUNDHyperglycemia has been shown to be a powerful predictor of poor outcome after ST-segment elevation myocardial infarction (STEMI). This study aimed to evaluate the effect of admission glucose on microvascular flow after successful primary percutaneous coronary intervention (PCI) in patients with STEMI.

METHODSSuccessful primary PCI was performed in 267 patients with STEMI. The maximum ST elevation of single electrocardiogram (ECG) lead before and 60 minutes after PCI was measured, and patients were then divided into 3 groups according to the degree of ST-segment resolution (STR): absent (<30%), partial (30% to 70%) or complete (> or =70%).

RESULTSOf the 267 patients, 48 (18.0%) had absent STR, 137 (51.3%) experienced partial STR, and 82 (30.7%) had complete STR. The degree of STR decreased with increasing admission glucose levels (P=0.032), and patients with hyperglycemia (serum glucose level > or =11 mmol/L) were more likely to have absent STR (P=0.001). Moreover,hyperglycemia was an independent predictor of incomplete STR (odds ratio, 1.870; 95% confidence interval, 1.038 to 3.371, P=0.037).

CONCLUSIONSHyperglycemia on admission is associated with abnormal coronary microvascular reperfusion in patients with STEMI after successful primary PCI, which may contribute, at least in part, to the poor outcomes in these patients.

Adult ; Aged ; Angioplasty, Balloon, Coronary ; methods ; Electrocardiography ; Female ; Glucose ; metabolism ; Humans ; Hyperglycemia ; blood ; pathology ; physiopathology ; Male ; Middle Aged ; Myocardial Infarction ; blood ; physiopathology ; therapy ; Odds Ratio

The effects of hypoxia on the level of reactive oxygen species (ROS), IkappaBalpha tyrosine phosphorylation, transcription of P65 mRNA and NF-kappaB activation in isolated rat peritoneal macrophages were investigated by DCFH-DA fluorescence spectrophotometry, Western blotting and RT-PCR. The results obtained are as follows. (1) During hypoxia, the levels of intracellular ROS began to increase at 1 h, then reached a peak at 2 h, and began to decrease after 3 h. IkappaBalpha tyrosine phosphorylation began to rise after 2 h hypoxia and was the highest after 3 h hypoxia. After 4 h hypoxia it decreased gradually. NF-kappaB activation began to increase after 3 h hypoxia, and reached a peak after 4 h hypoxia. (2) When antioxidant NAC (500 mmol/L) was added into the medium, the level of IkappaBalpha phosphorylation showed no significant changes during hypoxia. After adding protein tyrosine kinase inhibitor genistein (200 micromol/L), NF-kappaB activation induced by hypoxia was blocked significantly. (3) The expression of p65 mRNA was also elevated markedly during hypoxia. These results suggest that hypoxia may lead to IkappaBalpha phosphorylation and NF-kappaB activation through intracellular ROS, and that the regulation of NF-kappaB activity may involve IkappaBalpha phosphorylation and the expressions of each subunit gene of NF-kappaB.
Animals ; Cell Hypoxia ; Cells, Cultured ; Macrophages, Peritoneal ; cytology ; physiology ; Mice ; NF-kappa B ; biosynthesis ; genetics ; physiology ; Phosphorylation ; RNA, Messenger ; biosynthesis ; genetics ; Reactive Oxygen Species ; analysis ; Signal Transduction