Detection of Grass Carp Hemorrhage Virus (GCHV) from Vietnam and Comparison with GCHV Strain from China

Grass carp plays an important role in small-scale aquaculture in Vietnam. However, a severe disease, known in Vietnam as "Red Spot Disease", is causing significant economic loss in grass carp aquaculture. In this study, the tissue samples isolated from the grass carp with Red Spot Disease in Vietnam are investigated and comparied with the control GCHV isolated in China by experimental infection, culture cell infection, serological cross reactivity, and RT-PCR amplification. Infected grass carp exhibits hemorrhage symptoms about 5 days after experimental injection with GCHV-V (Vietnam) strain. The symptoms and lethality induced by the GCHV-V strain are identical to that induced by the Chinese GCHV-9014 strain. The Chinese GCHV-873 strain induces typical cytopathogenic effects in 4 cell lines, such as CIK, CAB, FHM and GCO, from the 6 fish cell lines examined. No cytopathogenic effects are observed in all the 6 examined cell lines, including CAB, FHM, CIK, EPC, CCO and GCO, infected by the GCHV-V strain and GCHV-9014 strain. Immunodiffusion assays demonstrate an obvious cross-reactivity among three GCHV strains. Precipitin lines are clearly observed not only between the anti-GCHV-873 serum and the two strains GCHV-873 and GCHV-9014, but also between the anti-GCHV-873 serum and the GCHV-V strain. GCHV can be detected by immunodiffusion assays after three generations of blind propagations in the cell lines inoculated by GCHV-V strain. This implicates that GCHV-V viruses have been replicated and amplified despite there being no cytopathogenic effects observed in these examined cell lines. Three genome segments of GCHV, including S8, S9 and S10, are amplified by three sets of PCR primers designed according to the segment sequences published recently. The Q8fp and Q8rp primer set specific for genome segment S8 amplifies a 955 bp fragment from the extracted sample of diseased fish with Red Spot Disease, and the fragment size is identical to that amplified by the same primer set from control GCHV-873 strain. Simultaneously, the Q9fp and Q9rp primer set specific for genome segment S9 generates a same 635 bp product, and the Q10fp and Q10rp primer set specific for genome segment S10 produces a same 697 bp fragment from both template samples of diseased fish with Red Spot Disease and control GCHV-873 strain. The RT-PCR amplification and corresponding size comparison data indicate that the three GCHV-V genome segments extracted from the diseased grass carp with Red Spot Disease in Vietnam should be identical to that in control GCHV-873 strain from China. The data confirm that the causative agent of grass carp Red Spot Disease in Vietnam is a virus, and the virus is closely similar to GCHV strain in China.

CONSTRUCTION OF EUKARYOTIC EXPRESSION VECTOR WITH GRANULOCYTE-MACROPHAGE COLONY-STIMULATING FACTOR GENE

Objective: To construct the eukaryotic expression vector that express human granulocyte-macrophage colony-stimulating factor (hGM-CSF) gene for making highly express in mammalian cells. Methods: Extract totally RNA from the induced human fetal lung (HFL) cell line. HGM-CSF cDNA was obtained by reverse transcription-polymerase chain reaction (RT-PCR), and then directionally subcloned into the HindIII and EcoRI site on the pcDNA3.1 plasmid, which was controlled by the CMV promoter, to form the recombinant expressing vector pcDNA3.1-GM-CSF. Results: The PCR amplification was identified and the sequence was analyzed, the results showed that hGM-CSF was properly inserted into the vector and the sequence was correct.

Abnormal expression of VEGF and its gene transcription status as diagnostic indicators in patients with non-small cell lung cancer

Objective Angiogenesis is known to be essential for the survival,growth,invasion,and metastasis of lung cancer cells. Vascular endothelial growth factor(VEGF) is an important factor regulating angiogenesis of non-small cell lung cancer(NSCLC); however,its pathologic features and significance are unclear. In this study,the tissue VEGF expression levels and its gene transcriptional status,as well as circulating VEGF levels,were investigated in patients with lung disease. Methods VEGF protein and m RNA expression levels in 38 lung tissue samples were analyzed by immunohistochemistry and reverse transcription-polymerase chain reaction(RT-PCR),respectively. Circulating VEGF levels were detected quantitatively by an enzyme linked immuno-sorbent assay. Results The level of VEGF expression was significantly higher in lung cancer tissue than in the corresponding paracancerous or non-cancerous tissues. The average level of VEGF-positive staining was 76% in tissue samples from NSCLC patients; the levels were 89% in tissue samples from stage III patients and 92% in stage IV patients. High VEGF expression was also evident in cases with lymph node metastasis(84%),distant metastasis(90%),and lower differentiation degree(89%). VEGF m RNA in cancerous tissues was represented predominantly by the VEGF121 and VEGF165 isoforms. Circulating VEGF levels were significantly higher in NSCLC patients [(840 ± 324) pg/m L] than in patients with benign lung diseases [(308 ± 96) pg/m L] or in healthy individuals serving as controls [(252 ± 108) pg/m L]. Conclusion The over-expression of lung VEGF and its gene transcription status should be useful molecular indicators for NSCLC diagnosis.

EXPRESSION AND SIGNIFICANCE OF PTEN IN ENDOMETRIAL CARCINOMA

Objective: To investigate the expression of PTEN in endometrial carcinoma and its clinical significance. Methods: Reverse transcriptase-polymerase chain reaction and Western-blot methods were used to detect PTEN expression in 28 cases of endometrial carcinoma. Results: mRNA and protein expression levels of PTEN in endometrial carcinomas were significantly lower than those in normal endometrium (P<0.01). Conclusion: PTEN may play an important role in the tumorigenesis of endometrial carcinoma.

不同类型淋巴瘤Survivin的表达及其意义

背景与目的:淋巴瘤的诊断与分型是临床病理诊断的难点。本研究检测抗凋亡基因survivin在不同类型淋巴瘤中的表达,并探讨其对淋巴瘤分型的意义。方法:用免疫组化法检测83例淋巴瘤、5例淋巴结反应性增生石蜡标本中survivin蛋白的表达;同时用RT-PCR检测K562、HL-60、Raji、Jurkat细胞系和以上病例中18例淋巴瘤及2例淋巴结反应性增生新鲜标本中survivinmRNA的表达;对不同类型的淋巴瘤survivin蛋白及mRNA表达进行半定量分析。结果:Survivin蛋白在非霍奇金淋巴瘤(non-Hodgkinslymphoma,NHL)的DLBL(diffuselargeBcelllymphoma)、BL(Burkittlymphoma)、LBL(lymphoblasticlymphoma)中有较高的表达,分别为87.2%(34/39)、100%(2/2)、85.7%(6/7),而在FL(follicularlymphoma)、MALT(extranodalmarginalzoneB-celllymphomaofmucosa-associatedlymphoidtissue)和MZL(marginalzonelymphoma)中表达较低,分别为22.2%、33.3%和40.0%,且多为弱阳性。高表达组(DLBL、BL、LBL)与低表达组(FL、MZL、MALT)之间差异有统计学意义(P<0.01)。并且DLBL中survivin阳性者中位年龄为57岁,明显高于阴性者41岁。霍奇金淋巴瘤(Hodgkinslymphoma,HL)中大部分R-S(Reed-Sternberg)细胞表达survivin蛋白。

Msi1基因沉默对人肝癌HepG2细胞的影响

目的:探讨siRNA沉默Msi1基因表达后,人肝癌HepG2细胞的增殖与凋亡的情况。方法:设计合成针对Msi1 mRNA序列的siRNA多条序列,分别转染人肝癌HepG2细胞。使用RT–PCR方法检测各组细胞Msi1基因表达情况;Western blot法检测survivin蛋白、caspase3蛋白表达情况;MTT法、平皿克隆实验检测HepG2细胞增殖情况;流式细胞术检测各组HepG2细胞凋亡情况。结果:(1)RT-PCR结果:转染后24 h后,序列a,b,c转染效率分别为65%,64%及62%。其抑制率分别是Msi1 siRNAa(68%)、Msi1 siRNAb(56%)、Msi1 siRNAc(35%)。(2)流式细胞术检测各组细胞凋亡:空白组、阴性组、脂质体组、Msi1 siRNAa组凋亡率分别为(4.14±0.26)%、(4.51±0.78)%、(4.19±1.21)%,(16.8±0.26)%,Msi1 siRNAa组与空白对照组比较差异有统计学意义(P<0.05)。(3)Msi1 siRNAa最能有效抑制肝癌HepG2细胞中Msi1表达,与空白对照组比较,Msi1 siRNAa组HepG2细胞生长、增殖速度明显减缓(P<0.05);survivin蛋白表达水平显著下调(P<0.05),而caspase3蛋白表达水平上调(P<0.05);Msi1 siRNAa组凋亡率增高(P<0.05)。结论:沉默Msi1可抑制人肝癌HepG2细胞的增殖,诱导其凋亡。

Isolation and Expression Analysis of MaPRMT1 Gene in Banana

[Objective] The aim of experiment was to lay molecular foundation for studying maturity mechanism of banana after harvest. [Method] The combined method of suppressing subtractive hybridization and cDNA micro-array were used to obtain cDNA segment of one PRMT gene in banana and the whole cDNA sequence of the gene was cloned.The bioinformatics analysis was operated on it,in addition, the expression profile analysis was conducted in different organs and different mature periods of banana.[Result] The whole length of cDNA in MaPRMT1 was 1 158 bp and possessed a complete open reading frame,which could encode 385 amino acids.It had high homology with PRMT in plant,containing one Methyltransf_1 domain.The MaPRMT1 gene was expressed in root,stem,leaf and fruit of banana and the expression levels in stem and leaf were relatively high.As the increase of days after harvest,the expression level declined gradually,however it reached maximum when ethylene release was biggest,then it declined.[Conclusion] MaPRMT1 belonged to the first kind of arginine methyltransferase and it was expressed differently in different organs and fruits at different mature periods.

Polypeptide Daintain as a New Biomarker for Detecting Breast Tumor

Daintain, a novel bioactive peptide produced and secreted by macrophages, was expressed in breast tumor tissues. The spatial distributions of daintain in 66 breast tumor specimens were investigated with immuno-histochemistry method. Reverse transcription polymerase chain reaction （RT-PCR） and in situ hybridization inspection system were also used to detect daintain in 45 cases of malignant breast tumors. The final results show that 93% high positive responses to daintain on breast cancer tumors. RT-PCR demonstrated that, no smear of daintain transcripted in benign tissues was found, and light smear in peri-cancer tissue was observed. Distribution of daintain was distinguishable among benign tissues, hyperplasia tissues, immature hyperplasia and invasive breast cancer, which can be used to mark the progression of the malignant lesion development. We conclude that the expression of daintain is up-regulated in breast cancers, which indicates that the peptide is closely associated with the disease progression. So daintain could be used as the biomarker for detecting breast cancer.

The detection of micrometastases in the peripheral blood of patients with breast cancer for hSBEM mRNA and CD44V6 mRNA

Objective： Successful treatment of breast cancer greatly depends on the early detection of its metastasis, therefore a sensitive and specific biomarker for detecting dissemination of the cancer cells will help to achieve this goal. This study was to evaluate the prognostic significance of human small breast epithelial mucin （hSBEM） and CD44V6 in breast cancer. Methods： The expressions of hSBEM mRNA and CD44V6 mRNA were detected with nested reverse transcription polymerase chain reaction （nested RT-PCR） in 67 samples of breast cancer and adjacent normal breast tissue, 16 samples of breast benign lesions tissue, and 67 specimens of peripheral blood from patients with breast cancer, 16 specimens of benign breast lesions, 20 specimens of healthy volunteers, and 25 （each five cases） other carcinomas tissue samples, including those of gastric carcinoma, colorectal carcinoma, esophageal carcinoma, lung carcinoma, and ovary carcinoma, were analyzed for hSBEM mRNA expression by nested RT-PCR. Results： hSBEM mRNA expression was observed in 62/67 （92.54%） of breast cancer, 14/16 （87.50%） of breast benign lesions and 59/67 （88.05%） of normal breast tissue, with no significant differences between them （P 〉 0.05）. None of the samples from other cancer tissues were positive. In peripheral blood the expression of hSBEM mRNA was detected in 34/67 （50.75%） from patients with breast cancer, with significant increasing （P 〈 0.05） in the cases of metastatic disease （stage Ⅳ） and those with lymph node metastasis compared with localized disease （stage Ⅰ, Ⅱ and Ⅲ） and without lymph node metastasis, but its expression was not found in peripheral blood of patients with benign breast lesions or healthy volunteers. Although CD44V6 mRNA was significantly higher in breast cancer than in benign breast lesions tissue and normal breast tissue, its expression in peripheral blood show no significant difference （P 〉 0.05） in the patients with breast cancer （82.09%）, benign breast lesion （75.00%）, or healthy volunteers （70.00%）. The expressions of hSBEM mRNA and CD44V6 mRNA had no correlation with the age of the patients, size of primary tumor, histological type and estrogen or progestin receptor status （P 〉 0.05）. Conclusion： hSBEM mRNA, as assessed by nested RT-PCR, shows a mammary-specific and mammary-sensitive expression, and is a sensitive indicator of hematogeneous spread of breast cancer cell, while CD44V6 shows low sensitivity and specificity in detecting dissemination of breast cancer cell in peripheral blood, hSBEM mRNA is a promising molecular biomarker for detecting breast cancer micrometastases.

Sequence Analysis and Quantitative Detection of Norwalk-like Viruses in Cultured Oysters of China

We isolated 4 Norwalk-like viruses （NLVs） contaminated oysters from 33 Chinese oysters collected from local commercial sources of Shandong Province. After amplification of the RNA-dependent RNA polymerase （RdRp） region of NLVs genomes with RT-PCR, the open reading frame 1 （ORF 1） of the RdRp was sequenced and subjected to multiple-sequence alignment. The results showed that NLVs in the four isolates belong to genogroup Ⅱ. The sequence comparison showed that the similarity between four Chinese oyster isolates were higher than 99.0%, which indicated that NLVs prevalent in close areas have high homogeneity in genome sequences. In addition, the most conserved sequences between diverse NLVs were used to design primers and TaqMan probes, then the real-time quantitative PCR assay was performed. According to the standard curve of GII NLVs, the original amounts （copies） of NLVs in positive patient＇s fecal isolate, positive Japanese oyster isolate, and the Chinese oyster isolate were 8.9× 10^8, 1.25× 10^8 and 4.7× 10^1 respectively. The detecting limit of NLVs was 1 × 10^1 copies. This study will be helpful for routine diagnosis of NLVs pathogens in foods and thus for avoiding food poisoning in the future.

Molecular characterization of yolk proteins in the female crab Neptunus pelagicus(A.Milne-Edwards,1861) from the Mediterranean Sea of Alexandria,Egypt

This study aimed to characterize the morphological changes in the ovary of the female crab Neptunus pelagicus and to identify specific fractions of vitelloginin and vitelline molecules during primary and secondary vitellogenesis.Samples of the blue crab were collected from the Mediterranean Sea of Alexandria monthly during 2017.Ovaries and oocytes in primary and secondary vitellogenesis were detached and treated for histological test.Native polyacrylamide gel electrophoresis(PAGE)Bis-Tris Gels was applied to identify vitelloginin(VN)and vitelline(VL)molecules.Protein Analyses were done by PAGE-SDS.The initial degenerate primers were built regarding the conserved amino acid domains of the yolk proteins.Primary and secondary vitellogeneses consisted of 8 phases.Lipoprotein fraction with molecular weight 550 kDa was identified in the hemolymph in secondary vitellogenesis.Two protein fractions(VLI&VLII)were identified in secondary vitellogenic oocytes.The electrophoresis performed with extract of stage I oocyte showed two protein fractions with molecular weights 550 kDa and 460 kDa.In stage II and III oocyte,4 subunits were presented of 180,195,140 and 120 kDa in VLI and 2 subunits with molecular weight of 110 kDa and 95 kDa in VLII.Another two fractions in stage V oocyte presented with molecular weights of 380 kDa and 360 kDa.Western blot analysis proved that both fractions were of four major polypeptide subunits with molecular weight of 180,125,90 and 85 kDa in each of the two VLs.The hybridization signal obtained by the Northern blot was detected in the hepatopancreas during ovarian cycle and in the ovary during secondary vitellogenesis.The result of the reverse transcription-polymerase chain reaction(RT-PCR)analysis showed that the mRNA that encodes the C-terminal region of the VN cDNA was found in the ovary in secondary vitellogenesis and in the hepatopancreas.

Aberrant Expression of TNF-α and TGF-β_1 mRNA in Spontaneous Abortion

Objective To investigate the aberrant expressions of TNF-α and TGF-β1 in peripheral blood mononuclear cells （PBMCs） and placental tissues in patients with early spontaneous abortion Methods Using the technique of semi-quantitative reverse transcript-polymerase chain reaction （RT-PCR）, TNF-α mRNA and TGF-β1 mRNA in PBMCs were measured in spontaneous abortion group （30 cases）, normal pregnancy group （25 cases） and nonpregnant group （25 cases）. The expressive intension of TNF-α protein and TGF-β1 protein in placental tissues was also identified by immunohistochemistry. Results Both levels of TNF-α mRNA and TGF-β1 mRNA expressed in PBMCs were significantly different between the three groups respectively （P〈0.05）. Levels of TNF-α in syncytiotrophoblastic and cytotrophoblastic cells of the two aborted groups were substantially higher than those of the nonregnant group （P〈0.01）, but the levels of TGF-β1 in syncytiotrophoblastic cells of the two aborted groups were markedly lower than those of the non-pregnant group （P〈0.01）. Conclusion There is potential relation between TGF-β at the fetomaternal interface and spontaneous abortion. TGF-β1 may contribute to the maintenance of pregnancy, and low-level expression of TGF-β1 may be associated with pregnancy failure.

OVEREXPRESSION OF Akt-1 GENE IN HUMAN HEPATOCELLULAR CARCINOMA

Objective: To investigate the expression difference of protein kinase B/Akt (Akt-1) between hepatocellular carcinoma (HCC) and adjacent normal liver tissues through the use of semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) and Northern blot. Methods: RT-PCR of 24 pairs of specimens and Northern blot of 4 pairs of specimens were performed to investigate the expression of Akt-1. Results: Akt-1 gene was overexpressed in 15 of 24 HCC (63.3%) by RT-PCR and in all HCC (4 paired tissues) by Northern blot. Conclusion: Akt-1 activation may play a role in the pathogenesis and progression of HCC. Akt-1 gene is reported to have changed in HCC for the first time. The precise relationship between Akt-1 and HCC is a matter of further investigation.

Predictive biomarker and clinicopathological characteristics analysis for esophageal cancer

Objective:The aim of this study was to explore the expression of manganese superoxide dismutase(MnSOD) in esophageal squamous cell carcinoma and its relationship with clinicopathological characteristics and its biological behavior.Methods:Immunohistochemical method(SP method),reverse transcription-polymerase chain reaction(RT-PCR) and Western blot were combined to detect the MnSOD protein and mRNA expression in 45 cases of esophageal squamous cell carcinoma and the normal tissue that was 5 cm apart from the edge of esophageal cancer lesion and without documented microscopic invasive cancer.Meanwhile,analysis was performed on the relationship between the pathological features of esophageal cancer and its biological behavior.Results:In esophageal squamous cell carcinoma and normal esophageal tissue,MnSOD protein expression was identified in 31.1%(14/45) and 86.7%(31/45)(P = 0.003),respectively,with the relative expression levels of MnSOD mRNA were 0.310 ± 0.036 and 0.482 ± 0.053(P = 0.000).The longer the lesions and the deeper the invasion,the differentiation would become poorer and the expression level of MnSOD would get lower,indicating that the level of MnSOD protein and mRNA expression were closely related to esophageal squamous cell carcinoma in the length of lesion,depth of invasion,and degree of differentiation(P < 0.05).Nevertheless,it showed no association with the presence of the lymph node metastasis,lesion site and the macroscopic classification(P > 0.05).Conclusion:The MnSOD protein and mRNA expression were both decreased in esophageal squamous cell carcinoma tissue.This may be related to the carcinogenesis and development of esophageal cancer.Detection of the expression of MnSOD would be of clinical significance in understanding the prognosis and guiding therapeutic strategy of esophageal cancer.

Effects of Mangiferin on the Expression of TNF-α,iNos,ICAM-1 and Its mRNA in the Heart,Brain and Kidneys of SHR

[Objectives] To study the effects of Mangiferin( MGF) on TNF-α,iNOS,ICAM-1 and its mRNA expression in the heart,brain and kidneys of spontaneously hypertensive rats( SHR),and reveal the mechanism of its anti-inflammatory injury in hypertension target organs.[Methods]SHRs were randomly divided into 5 groups: the model group,the high-dose,medium-dose,low-dose MGF groups and the Benazepril group,with 8 rats in each group,WKY rats were used for the normal control group. Besides,on-invasive blood pressure( BP) instruments were used to measure systolic blood pressure in the rats' tail artery,western blot was used to analyze the expression of TNF-α,iNOS,ICAM-1 and reverse transcription-polymerase chain reaction( RT-PCR) was used to analyze the expression of TNF-α,iNOS,ICAM-1 mRNA.[Results]Compared with the normal control group,the model group's BP level was significantly increased( P <0. 01)),but the MGF had no significant lowering BP effect( P > 0. 05); compared with the normal control group,the expression of TNF-α,iNOS,ICAM-1 and its' mRNA in the model group was significantly increased( P < 0. 05 or P < 0. 01),and MGF could reduce the level of expression of these inflammatory cytokines( P < 0. 05 or P < 0. 01); between the Benazepril group and high-dose,medium-dose,low-dose MGF groups,most of the indicators had no significant difference( P > 0. 05). [Conclusions]MGF had no significant lowering BP effect,SHR showed inflammatory injury in the heart,brain and kidneys,MGF showed improvement on the inflammatory injury,and the anti-inflammation mechanism may be associated with lowering TNF-α,i NOS and ICAM-1 and its mRNA expression.

Detection of CEA mRNA in patients with non-small cell lung cancer and it’s significance

Objective: To detect the expression of CEA mRNA in patients with non-small cell lung cancer (NSCLC) and to investigate it's significance. Methods: The blood samples were taken from peripheral veins of 70 patients with NSCLC and 18 patients with benign diseases at 3 intervals during the surgery. The transcription of carcinoembryonic antigen messenger ribonucleic acid (CEA mRNA) was assayed by means of nested reverse transcriptase polymerase chain reaction (RT-PCR) and micro-fluid chip. Results: The CEA mRNA positive rates at each of the 3 time spots were as follows: 50% at beginning of the surgery (group 1), 62.8% in the samples collected when ligating the pulmonary vein (group 2) and 57.1% in samples collected 1 h after ligation (group 3). A significant difference was found between groups 1 and 2 (χ2 = 7.114, P < 0.05). Con-clusion: Cancer cell dissemination during surgery is demonstrated indirectly in our study, when to ligate the pulmonary vein (earlier or later) may affect the quantity of tumor cells spread into the circulation.

Callus Cultures Of Beans Infected With Virus As A Model For Testing Antiviral Compaunds

In the work,bean callus raised from a leaves of Bean common mosaic virus infected bean plant was obtained and adapted for the testing of antiviral activity of liposomal glycan-glycolipid complexes.Ganoderma adspersum glucans and Pseudomonas spec.rhamnolipids were constituents of liposomal compaunds.It has been shown that under the long-term cultivation(up to 3 months)in the presence of a liposomal preparation containing(10-100 mg/l),the virus is eliminated from the tissue.This is evidenced by the absence of 391 bp sequence amplification product established by RT-PCR in the callus tissue,cultured on a medium containing the liposomal complex.The proposed model system is analogous to plant tumors and has obvious advantages over similar systems in vivo,since the callus growth is controlled and independent of environmental factors.

A novel cytogenetic abnormality r(7)(::p11.2->q36.3::) in a Philadelphia-positive chronic myeloid leukemia case

The so-calledPhiladelphia(Ph) chromosome is present in more than 90% of chronic myeloid leukemia (CML) cases. It results in juxtaposition of the 5' part of the BCR gene on chromosome 22 and the 3' part of the ABL1 gene on chromosome 9. An additional acquired monosomy 7 or deletion of 7q is associated with poor prognosis in a variety of myeloid disorders. Here we report a novel Ph chromosome positive CML case with a ring chromosome 7 [r(7)]. Immunophenotyping was compatible with CML, although 4.5% of total leucocytes appeared like acute myelogeneous leukemia (AML) subtype M2. The r(7) was characterized in detail by array-proven multicolor banding (aMCB), the latter being of enormous significance to characterize breakpoint regions in detail. Underlying mechanisms and prognostic are discussed, as ring chromosomes are rare cytogenetic abnormalities in hematopoietic malignancies.

Detection of respiratory viral and bacterial pathogens causing pediatric community-acquired pneumonia in Beijing using real-time PCR

Objective: The aim of this study was to determine the etiology and prevalence of pediatric CAP in Beijing using a real-time polymerase chain reaction (PCR) technique. Methods: Between February 15, 2011 and January 18, 2012, 371 pediatric patients with CAP were enrolled at Beijing Children's Hospital. Sixteen respiratory viruses and two bacteria were detected from tracheal aspirate specimens using commercially available multiplex real-time reverse transcription PCR (RT-PCR) kits. Results: A single viral pathogen was detected in 35.3%of enrolled patients, multiple viruses in 11.6%, and virus/bacteria co-infection in 17.8%. In contrast, only 6.5%of patients had a single bacterial pathogen and 2.2%were infected with multiple bacteria. The etiological agent was unknown for 26.7% of patients. The most common viruses were respiratory syncytial virus (RSV) (43.9%), rhinovirus (14.8%), parainfluenza virus (9.4%), and adenovirus (8.6%). In patients under three years of age, RSV (44.6%), rhinovirus (12.8%), and Streptococcus pneumoniae (9.9%) were the most frequent pathogens. In children aged 3e7 years, S. pneumoniae (38.9%), RSV (30.6%), Haemophilus influenzae (19.4%), and adenovirus (19.4%) were most prevalent. Finally in children over seven years, RSV (47.3%), S. pneumoniae (41.9%), and rhinovirus (21.5%) infections were most frequent. Conclusions: Viral pathogens, specifically RSV, were responsible for the majority of CAP in pediatric patients. However, both S. pneumoniae and H. influenzae contributed as major causes of disease. Commercially available multiplexing real-time PCR allowed for rapid detection of the etiological agent. Copyright ? 2015, Chinese Medical Association Production. Production and hosting by Elsevier B.V. on behalf of KeAi Communications Co., Ltd. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

Livin在原发性肝癌中的表达及与Caspase-3的关系

目的 研究凋亡抑制基因Livin在原发性肝癌中的表达,并探讨其与Caspase-3表达的相关性.方法 采用逆转录多聚酶链式反应(RT-PCR)方法检测Livin和Caspase-3在46例原发性肝癌,28例癌旁组织,12例正常肝组织中的表达,分析二者之间相关性.结果 Livin基因在肝癌组织、癌旁组织、正常肝组织表达依次降低,其表达与原发性肝癌患者的年龄、性别、肿瘤大小、是否合并乙肝肝硬化、分化程度、甲胎蛋白等差异均无统计学意义(P＞0. 05),而与肝癌是否发生转移有关(P<0. 05);Caspase-3在上述组织中依次升高,差异均有统计学意义,二者在肝癌组织中表达负相关(rs=-0. 333, P<0.05).结论 Livin在肝癌组织中高表达,可能和Caspase-3在原发性肝癌细胞凋亡中起重要作用,可能是原发性肝癌生物学行为和预后重要标记物之一.