Objective To explore the effect of carvedilol on ACE2 gene and protein expression in chronic heart failure rats after myocardial infarction.Methods The heart failure model was induced by acute myocardial infarc- tion (AMI) through ligating the left anterior descending coronary artery.One month after operation,rats were randomized to receive placebo or carvedilol 2 mg/(kg?d),by gavage.Sham-operated rats were used as the control group.Hemodynamies,body mass and left ventrieular mass index,plasma and myocardial level of angiotensin Ⅱ were determined.ACE2 gene and protein expression was assessed by using RT-PCR and Western Blot.Results The mortality of placebo and Carvedilol groups were 20%,compared with 0% in sham operated rats.Carvedilol significantly improved LVEDP,LVSP,+dp/dt_(max) and-dp/dt_(min) in CHF rats but all the hemodynamics data were still inferior than that of controls.Plasma and myocardial angiotensin Ⅱ level were increased significantly in CHF placebo rats than those of control rats (plasma Ang Ⅱ:CHF:194?19 vs controls:132?15 ng/L,myocardium Ang Ⅱ:CHF:6.7?0.4 vs control:3.8?0.3 ng/g,P

OBJECTIVE:To investigate the preventive and therapeutic effect of Shuganning injection on alcoholic liver fibrosis (ALF) in model rats,and provide experimental basis for its clinical application for alcoholic liver disease. METHODS:50 rats were enrolled and intraperitoneally given mixed liquid of 60% alcohol-corn oil-pyrazole to reduce ALF model. Another 10 rats were enrolled and intraperitoneally given normal saline,as normal control group. After 16 weeks,survived model rats(n=40)were ran-domly divided into model group,positive control group(Anluo huaxian pill 0.75 g/kg,ig),Shuganning injection high-dose,medi-um-dose,low-dose groups(4.8,2.4,1.2 mL/kg,ip),8 in each group. Normal control group and model group were intraperitone-ally injected equal volume of normal saline (5 mL/kg),administration groups were given relevant medicines,once a day,for 8 weeks;and modeling was contiuously conducted at the same time. After administration,body mass of rats was weighed,and the levels of liver function indexes [aspartate aminotransferase(AST),alanine aminotransferase(ALT)] and liver fibrosis indexes [hyal-uronic acid(HA),laminin(LN),type Ⅲ procollagen(PⅢNP),type Ⅳ collagen(Ⅳ-C)] in serum of rats were detected. Liver index of rats was determined and pathological changes of liver tissue were observed. RESULTS:Compared with normal control group,body mass of rats in model group was significantly decreased(P<0.05);liver index,and liver function index,liver fibro-sis index levels in serum were significantly increased (P<0.05 or P<0.01). Liver tissue showed steatosis,hepatocyte vacuoliza-tion,a large number of fibrous tissue deposition around portal areas and other pathological changes. Compared with model group,above-mentioned changes were improved significantly in administration groups (P<0.05 or P<0.01). CONCLUSIONS:Shugan-ning injection can obviously improve liver tissue damage of model rats with ALF,showing certain preventive and therapeutic effect on alcoholic liver disease.

Objective To evaluate the risk factors of developing post-engraftment hemorrhagic cystitis (HC) in patients receiving allogeneic stem cell transplantation (allo-HSCT).Methods Retrospective data was collected from 92 patients with acute leukemia (acute myeloid leukemia 41 and acute lymphoblastic leukemia 51) who underwent allo-HSCT from 2000 to 2010,and the association of pre-transplantation parameters with the incidence of post-engraftment HC was analyzed.Results Forty-three patients had HLA-matched donors and 49 had unrelated donors.Of these patients,25 developed HC at a median of 35 days (day +20 to +65) after allo-HSCT.In the univariate analysis,unrelated donor,gender mismatch (female donor to male recipient),conditioning containing busulfan,graft-versus-host disease (GVHD),prophylaxis with cyclosporine (CSA) + methotrexate (MTX) + mycophenolate mofetil (MMF),use of anti-thymoglobulin (ATG) and development of GVHD were associated with increased incidence of HC.In the multivariate study,gender mismatch (P =0.001),use of ATG (P ＜ 0.001),and GVHD (P =0.007) remain as independent factors for the increased risk of HC.More importantly,with these 3 factors,it is able to classify patients into 4 groups with risk of postengraftment HC at (7.7±4.6) ％,(22.9±7.1) ％,(48.2±10.5) ％,and 100.0 ％,respectively.Conclusion This retrospective study identified the gender mismatch,use of ATG in the preparation regimen,and aGVHD as important risk factors to predict the development of post-engraftment HC.Based on these risk factors,it is possible to classify patients into different risk groups for post-engraftment HC.Prospective study with a large cohort of patients is warranted to confirm the findings.Future clinical trial for HC prevention and treatment must be carried out on the intermediate and high-risk patients.

Purpose To investigate the positive rate and concordance rate of BRAF mutation in papillary thyroid carcinoma detected by real-time PCR method and Sanger sequencing. Methods 312 papillary thyroid carcinomas patients were enrolled in this study. Real-time PCR method and Sanger sequencing were performed to detect BRAF gene mutations. The frequency of BRAF mutation and the concordance of two methods were analyzed. Results BRAF mutation was detected in 65. 4% (204/312) and 63. 8% (199/312) of 312 papillary thyroid carcinoma samples by using real-time PCR method and Sanger sequencing, respectively. There was no significant correlation between BRAF gene mutations and patients’ gender. There was significant correlation between BRAF gene mutations and patients’ age. The overall concordance between real-time PCR method and Sanger sequencing for BRAF mutation detection was 98. 4%. Conclusion Real-time PCR method provides an effective method in BRAF gene mutation detection.

PURPOSE: The purpose of this study was to test the reliability and validity of the Behavior Assessment for Children (BAC) in a community of school-aged children in Taiwan. METHOD: A school-based sample comprising third grade and fourth grade students was recruited from Taichung City in Taiwan. The parents (n = 248) and teachers (n = 15) of these students completed structured questionnaires, including the Child Behavior Checklist (CBCL) and the proposed BAC. Content validity, concurrent validity, construct validity, internal consistency, and inter-rater reliability of the BAC were assessed. RESULTS: The BAC comprised three subscales (attention, emotion, and self-control) that included 17 items. The content validity index (CVI) score was 0.98. The result of the confirmatory factor analysis (goodness of fit = .90, root mean square of residual = .03, root mean square error of approximation = .06, and comparative fit index = .94) supported the construct validity of the three BAC subscales. The concurrent validity of the BAC subscales significantly correlated with the compatible CBCL subscales (r = .59-.78, p < .001). Cronbach α of the subscales of the BAC ranged from .78 to .92. The intraclass correlation coefficient between the parents and teachers ranged from .31 to .44, and the joint probability of agreement ranged from 31.4% to 92.2%. CONCLUSIONS: The BAC is a valid and reliable instrument for evaluating behavioral problems in schoolaged children.
*Attention ; Child ; Child Behavior Disorders/*diagnosis ; *Diagnostic Techniques and Procedures ; *Emotions ; Female ; Humans ; Male ; *Psychometrics ; Reproducibility of Results ; *Self-Control ; Taiwan

Tumor immune checkpoint therapy is a clinical treatment strategy developed based on the new principle of the inhibition of negative immune regulation. In this article, the tumor immune checkpoint therapy and the drug delivery strategies were reviewed, mainly including immunity and tumor therapy, tumor immune checkpoint therapy and its mechanism of action, clinical application of tumor immune checkpoint therapy and therapeutic drugs, immune resistance of programmed cell death protein 1 (PD1)/programmed cell death ligand 1 (PDL1) treatment and countermeasures, drug delivery strategies for tumor immune checkpoint therapeutic agents, etc. As a revolutionary new immunotherapy strategy, tumor immune checkpoint therapy has shown obvious superior therapeutic efficacy in a variety types of tumor. However, tumor immune checkpoint therapy is also faced with a big challenge, namely, immunotherapy resistance. With the discovery of new mechanism, the continuous development of new therapeutic drugs and delivery strategies, tumor immune checkpoint therapy is expected to further improve the clinical efficacy of tumor.

OBJECTIVE: To explore the mutation of Y-STR loci in meiotic allelic transmission in a large pedigree. METHODS: The oral swabs of 163 male individuals were collected from a Lin pedigree. Twenty-two Y-STR genetic markers were typed with AGCU Y24 fluorescent detection kit (AGCU Y24 system), which also contained 16 Y-STR markers included in Yfiler multiple amplification kit (Yfiler system). The genotyping results of Y-STR loci were compared between each two males in the pedigree. RESULTS: There were 20 and 30 kinds of haplotypes obtained with Yfiler and AGCU Y24 systems in 163 male individuals from the Lin pedigree, respectively. The rates referred to haplotype differences (RRHD) of these two typing systems between male pairs were 0.910 5 and 0.922 7, respectively. The average number of marker differences were 6.582 1 and 9.824 8, respectively. The RRHD increased along with the incidents of meiosis. CONCLUSION Y-STR mutation leads to different Y-STR haplotypes among the male members in a paternal pedigree and the rate of difference increases along with the incidents of meiosis.
Alleles ; Chromosomes, Human, Y/genetics* ; DNA Fingerprinting ; Genetic Linkage ; Genetic Markers/physiology* ; Genotype ; Haplotypes ; Humans ; Male ; Mutation/genetics* ; Pedigree

OBJECTIVETo investigate the feasibility of real-time PCR-optimized oligonucleotide probe method for detection of KRAS mutations in lung and colorectal carcinomas, as compared with Sanger sequencing method.

METHODSGenomic DNA was extracted from formalin fixed paraffin embedded samples of 221 lung carcinomas and 131 colorectal carcinomas after tumor cell content assessment and macrodissection. Real-time PCR-optimized oligonucleotide probe method and Sanger sequencing were performed to detect KRAS gene mutations. The frequency of KRAS mutation, mutation types, and their concordance were analyzed.

RESULTSKRAS mutation was detected in 6.3% (14/221) and 4.5% (10/221) of 221 lung cancer samples by using real-time PCR-optimized oligonucleotide probe method and Sanger sequencing, respectively, while in 41.2% (54/131) and 40.5% (53/131) of 131 colorectal cancer samples, respectively. There was no significant correlation between KRAS gene mutations and patients' gender and age (P > 0.05). The positive rate of KRAS codon 12 mutation was significantly higher than that of KRAS codon 13 mutation (P < 0.05). The overall concordance between real-time PCR-optimized oligonucleotide probe method and Sanger sequencing for KRAS mutation detection was 97.4%.

CONCLUSIONReal-time PCR-optimized oligonucleotide probe method provides an alternative method with high consistency and sensitivity as compared to Sanger sequencing in gene mutation detection.

Codon ; Colorectal Neoplasms ; genetics ; DNA Mutational Analysis ; methods ; Female ; Humans ; Lung Neoplasms ; genetics ; Male ; Middle Aged ; Mutation ; Oligonucleotide Probes ; Proto-Oncogene Proteins ; genetics ; Proto-Oncogene Proteins p21(ras) ; Real-Time Polymerase Chain Reaction ; methods ; Sensitivity and Specificity ; ras Proteins ; genetics

OBJECTIVETo investigate the mutation frequencies of KRAS, BRAF, PIK3CA and EGFR genes that were effective on the targeted therapies in colorectal carcinoma.

METHODSThe tissue specimens from 331 colorectal cancer patients were collected and subject to KRAS, BRAF, PIK3CA and EGFR mutation analysis. Paraffin-embedded tissue samples were obtained and macrodissection was performed to enrich the tumor cells for DNA extraction when necessary. PCR-based direct DNA sequencing was used to investigate the codons 12 and 13 in exon 2 of KRAS gene, exons 11 and 15 of BRAF gene, exons 9 and 20 of PIK3CA gene and exons 18-21 of EGFR gene.

RESULTSActivating mutations were detected in KRAS (44.1%, 137/311), BRAF (5.8%, 9/156), PIK3CA (2.6%, 4/156) and EGFR (1.3%, 2/156) in the study cohort of colorectal carcinoma cases. Among KRAS gene mutations, 81.0% (111/137) occurred in codon 12, with p.G12D as the most common variant (45.3%, 62/137); 19.0% (26/137) occurred in codon 13, with 38G > A (G13D) as the most common variant (17.5%, 24/137).

CONCLUSIONSThe KRAS mutation frequency is the highest among the four genes (KRAS, BRAF, PIK3CA and EGFT) tested in colorectal carcinoma. The presence of these gene mutations may provide therapeutic information for targeted therapy. Mutation analyses of BRAF and PIK3CA in addition to KRAS should be a part of the standard diagnostic algorithm for colorectal carcinoma patients.

Class I Phosphatidylinositol 3-Kinases ; Codon ; Colorectal Neoplasms ; genetics ; pathology ; DNA Mutational Analysis ; Exons ; Female ; Humans ; Male ; Middle Aged ; Mutation ; Phosphatidylinositol 3-Kinases ; genetics ; Proto-Oncogene Proteins ; genetics ; Proto-Oncogene Proteins B-raf ; genetics ; Proto-Oncogene Proteins p21(ras) ; Receptor, Epidermal Growth Factor ; genetics ; ras Proteins ; genetics

AIMTo investigate the role of ICAM-1 on the surface of vascular endothelial cell (VEC) in freezing/thawing injury of VEC, in order to elucidate the pathogenesis of freezing/thawing injury.

METHODSVEC separated and cultured from rat aorta and PMN separated from rat peripheral blood were selected as experiment materials. The frozen/thawed VEC model was founded by freezing VEC with the type WKL-V rate cooling instrument and then rewarming them in a water bath. ICAM-1 expression on the surface of frozen/thawed VEC was detected at 4, 12 and 24 h after freezing/thawing with immunohistochemical method. After coincubating frozen/thawed VEC with normal PMN, the adhesion of VEC to PMN was monitored with rose bengal staining assay and the injury level of VEC was indicated by measuring LDH activity in nutrient solution.

RESULTSThe ICAM-1 expression on the surface of VEC increased from 13.2% +/- 3.6% before freezing/thawing of VEC to 22.3% +/- 4.4% at 4 hour after freezing/thawing, and reached the peak (37.9% +/- 2.5%) at 12 hour after freezing/thawing of VEC. After coincubation of frozen/thawed VEC with normal PMN, the adherence of frozen/thawed VEC to PMN increased from group control 0.204 +/- 0.025 to 0.363 +/- 0.022 (P < 0.01), LDH activity in nutrient solution increased from group control 104.64 +/- 20.14 U/L to 162.33 +/- 27.88 U/L (P < 0.01), monoclonal antibody against ICAM-1 (ICAM-1 Mab) could partially block the adherence of frozen/thawed VEC to PMN (0.270 +/- 0.021, P < 0.01), and diminish LDH activity in nutrient solution (125.39 +/- 22.26 U/L, P < 0.05).

CONCLUSIONThe freezing/thawing of VEC can elicit an increase in ICAM-1 expression on the surface of VEC, and then proceed to VEC-PMN adherence and lead to VEC injury.

Animals ; Cells, Cultured ; Endothelial Cells ; metabolism ; Endothelium, Vascular ; cytology ; Freezing ; Intercellular Adhesion Molecule-1 ; metabolism ; Neutrophils ; cytology ; Rats