Hepatitis C virus in human B lymphocytes transformed by Epstein-Barr virus in vitro by in situ reverse transcriptase-polymerase chain reaction

AIM: To study persistence and replication of hepatitis C virus (HCV) in patients' peripheral blood mononuclear cells (PBMC) cultured in vitro. METHODS: Epstein Barr virus (EBV) was used to transform the hepatitis C virus from a HCV positive patient to permanent lymphoblastoid cell lines (LCL). Positive and negative HCV RNA strands of the cultured cells and growth media were detected by reverse transcriptase-polymerase chain reaction (RT-PCR) each month. Core and NS5 proteins of HCV were further tested using immunohistochemical SP method and in situ RT-PCR. RESULTS: HCV RNA positive strands were consistently detected the cultured cells for one year. The negative-strand RNA in LCL cells and the positive-strand RNA in supernatants were observed intermittently. Immunohistochemical results medicated expression of HCV NS3 and C proteins in LCL cytoplasm mostly. The positive signal of PCR product was dark blue and mainly localized to the LCL cytoplasm. The RT-PCR signal was eliminated by overnight RNase digestion but not DNase digestion. CONCLUSION: HCV may exist and remain functional in a cultured cell line for a long period.

DETECTION OF BREAST CANCER MICROMETASTASES IN BONE MARROW USING REVERSE-TRANSCRIPTASE CHAINREACTION AND SOUTHERN HYBRIDIZATION

Objective: The aim of this study was to detect micrometastases in bone marrow of primary breast cancer patients, and compare with other clinical parameters. Methods: Cytokeratin 19 (CK-19) gene mRNA expression was detected by reverse transcriptase-polymerase chain reaction (RT-PCR) and Southern blot hybridization. Human breast cancer cell line T47D was mixed with bone marrow cells in different proportions. The positive detection rate was compared among RT-PCR, Southern blotting and immunohistochemistry (IHC) methods. Results: Cytokeratin 19 gene was expressed in all 6 positive control samples while the expression was not seen in 8 negative control samples. In all 54 patients 14 cases were CK-19 positive (25.9%) by RT-PCR, another positive signal was obtained in 5/54 (9.3%) of bone marrow samples by Southern blotting. The total positive cases are 19/54 (35.2%). CK-19 IHC+ cells were detected at a dilution of one T47D cell in 5×104 bone marrow cells, while the sensitivity detected by PCR and Southern blot hybridization was at 1∶5×105 and 1∶1×106, respectively. This demonstrates that RT-PCR and Southern blotting was at least 20 times more sensitive than the IHC method. The micrometastases positive rate of the larger tumor size group (>5.0 cm) was significantly (P<0.05) greater than that of the smaller tumor size group (0–2.0 cm). Conclusion: detection of micrometastases in bone marrow by RT-PCR and Southern blotting, using CK-19 as a biological marker, is highly sensitive and it is a method to be used for anticipating the prognosis of breast cancer patients.

COMBINED DETECTION OF BREAST CANCER MICROMETASTASES IN THE LYMPH NODES AND BONE MARROW USING REVERSETRANSCRIPTASE CHAIN REACTION AND SOUTHERN HYBRIDIZATION

Objective: The presence of lymph nodes and bone marrow micrometastases of patients with breast carcinoma by immunohistochemistry (IHC) methods has been strongly correlated to early recurrence and shorter overall survival. The aim of this study was to detect micrometastases in matched sample pairs of lymph nodes and the bone marrow of primary breast cancer patients using a more sensitive method, and compare with other clinical parameters. Methods: Cytokeratin 19 (CK-19) gene mRNA expression was detected by reverse transcriptase-polymerase chain reaction (RT-PCR) and Southern blot hybridization. Human breast cancer cell line T47D was mixed with bone marrow cells at different proportions. The positive detection rate was compared among RT-PCR, Southern blotting and IHC methods. Results: Cytokeratin 19 gene was expressed in all 6 positive control samples, while the expression wasn’t seen in 18 negative control samples. CK-19 IHC positive cells were detected at a dilution of one T47D cell in 5×105 bone marrow cells, while the sensitivity detected by PCR and Southern blot hybridization was at 1:5×104 and 1:106, respectively. In the samples from the 35 patients, we found CK-19 positive cells in 2 cases (5.7%) by IHC. CK-19 gene expression signal was detected in 14/35 (40%) by RT-PCR, and 17/35 (48.6%) by southern blotting. Four cases were micrometastases positive both in lymph node and bone marrow (11.4%). There was no correlation between CK-19 detection and other clinical parameters. Conclusion: combined detection of micrometastases in lymph node and bone marrow by RT-PCR and Southern blotting, using CK-19 as a biological marker, is a highly sensitive method for breast cancer.

SEQUENCE ANALYSIS OF THE NS5 REGION OF GBVC/HGV AND DETECTION OF THE VIRUS BY REVERSE TRANSCRIPTASE PCR

GBV C/HGV is a newly identified virus associated with human hepatitis In this study, the nucleotide sequences of the partial NS5 gene of GBV C/HGV derived from sera of 8 Chinese patients were determined The nucleotide homology among the 8 isolates were 92% on average On the basis of sequence analysis, two sets of oligonucleotide primers derived from highly conserved region of GBV C/HGV NS5 gene were designed to establish both sensitive and specific nested PCR for detection of GBV C/HGV RNA 253 Chinese patients were examined for the virus RNA GBV C/HGV RNA positive rates in patients infected with HBV, HCV and patients with chronic non B,non C hepatitis were 18 4%, 19 8% and 8 9% respectively This result suggested that HBV,HCV and GBV C/HGV shared the same transmission risk factors 8 patients with GBV C/HGV and HCV coinfection were retrospectively observed for the response to interferon Coinfection with GBV/HGV did not negatively influence the responsiveness of HCV, and GBV C/HGV was sensitive to interferon to a certain degree

Detection of SYT-SSX fusion transcripts in paraffin-embedded tissues of synovial sarcoma by reverse transcription-polymerase chain reaction

OBJECTIVE: To assess the feasibility of detecting SYT-SSX fusion transcripts in paraffin-embedded tissues of synovial sarcoma by reverse transcription-polymerase chain reaction (RT-PCR). METHODS: RT-PCR was used to amplify the SYT-SSX fusion transcripts using archival formalin-fixed paraffin-embedded tumor specimens from a series of 37 synovial sarcoma cases. To investigate the specificity of the SYT-SSX fusion transcripts, a variety of non-synovial sarcoma tumors were included in the study as negative controls. The detected messages derived from fusion genes were confirmed by subsequent sequence analysis. RESULTS: SYT-SSX fusion transcripts were detected in 33 of 37 (89.2%) synovial sarcomas. None of the 34 cases of non-synovial sarcoma tumors showed amplified products of SYT-SSX fusion transcripts, although PBGD mRNA was detected in all specimens. Among 33 SYT-SSX-positive synovial sarcomas, 22 tumors had an SYT-SSX 1 fusion transcript, whereas 6 tumors had an SYT-SSX2 fusion transcript. Fusion types can not be distinguished in the remaining 5 cases. There was a significant relationship between SYT-SSX fusion type and histologic subtype. All 10 biphasic synovial sarcomas had the SYT-SSX1 fusion, whereas all tumors with SYT-SSX2 were of monophasic morphology (P

新型冠状病毒Omicron I1566V突变位点MS2噬菌体病毒样颗粒的构建

目的构建新型冠状病毒Omicron I1566V突变位点MS2噬菌体病毒样颗粒。方法选取并合成带有6×His标签的MS2噬菌体的衣壳蛋白(CP)、成熟蛋白(A蛋白)及Omicron I1566V突变基因序列,插入pACYCDuet-1质粒构建重组载体,通过原核系统诱导表达目的蛋白,纯化重组蛋白后利用透射电镜对蛋白质进行物理表征,最后通过反转录聚合酶链反应(RT-PCR)检测病毒样颗粒的热稳定性及耐核酸酶水解能力。结果成功构建包含有6×His标签的CP、A蛋白和Omicron I1566V突变基因序列的重组载体,经限制性内切酶BamHⅠ和KpnⅠ酶切鉴定和测序验证,结果均与预期相符。经诱导并纯化后,通过电镜观察到了大小均匀、直径为23~28 nm的病毒样颗粒,该病毒样颗粒经核酸酶消化后可在37℃条件下稳定储存20 d以上。结论该研究成功利用MS2噬菌体的CP和A蛋白构建了Omicron I1566V突变位点病毒样颗粒,为该突变位点的RT-PCR检测体系提供了可靠的质量保障。

Epidemiological and Subtype Characterization of Influenza Viruses Infection in Children in Shenzhen, China during Three Consecutive Seasons (January 2016-December 2018)

Background: Children with seasonal influenza infection cause a significant burden of disease each year in the pediatric clinic. Influenza A and B viruses are the major types responsible for illness. A better understanding of the periodicity facilitates the prevention and control of influenza in children. Objective: This study aims to analyze the epidemiological patterns and subtype characterization of influenza viruses among children in Shenzhen, China. Methods: Influenza samples were collected by nasopharyngeal swabs from influenza like illness patients in Shenzhen Children’s Hospital from January 2016 to December 2018. The positive cases and influenza subtypes were determined by gold labeled antigen detection and reverse transcriptase polymerase chain reaction. The influenza periodicity and age, subtype distribution as well as the association between climate parameters and different influenza subtypes were analyzed by SPSS 22.0. Results: The influenza positive rate during 2016-2018 was 21.0%, with a highest positive rate in the year 2018. The positive rate varied by month, season, and year describing a sequence of peaks presenting primarily in all year including spring, summer and winter. The characteristics of influenza peak were different in each year, with a spring peak in 2016 and a summer plus a winter-spring peaks in 2017 and 2018. In addition, influenza B exhibited a winter-spring seasonal pattern while influenza A displayed a more variable seasonality, highlighting influenza B rather than influenza A which had a negative association with climate parameters. Influenza-positive cases were older than influenza-negative cases (P P Conclusion: Influenza activity in children from Shenzhen typically displays both winter-spring and summer peaks. Influenza A epidemic occurred separately or co-circulated with influenza B, with a winter-spring pattern for influenza B and a much more variable seasonality for influenza A. Influenza B had a negative association with climate parameters. In addition, hospitalization with influenza often occurs in younger individuals infected with influenza A.

Comparison of gene expression profiles between primary tumor and metastatic lesions in gastric cancer patients using laser microdissection and cDNA microarray

AIM. To study the differential gene expression profiles of target cells in primary gastric cancer and its metastatic lymph nodes using laser microdissection （LMD） in combination with cDNA microarray. METHODS： Normal gastric tissue samples from 30 healthy individuals, 36 cancer tissue samples from primary gastric carcinoma and lymph node metastasis tissue samples from 58 patients during gastric cancer resection were obtained using LMD in combination with cDNA microarray independently. After P27-based amplification, aRNA from 36 of 58 patients （group 1） with lymph node metastasis and metastatic tissue specimens from the remaining 22 patients （group 2） were applied to cDNA microarray. Semiquantitative reverse transcriptase polymerase chain reaction （RT-PCR） and imrnunohistochemical assay verified the results of microarray in group 2 and further identified genes differentially expressed in the progression of gastric cancer. RESULTS： The expression of 10 genes was up-regulated while the expression of 15 genes was down-regulated in 22 gastric carcinoma samples compared with that of genes in the normal controls. The results were confirmed at the level of mRNA and protein, and suggested that four genes （OPCML, RNASE1, YES1 and ACK1） could play a key role in the tumorigenesis and metastasis of gastric cancer. The expression pattern of 3 genes （OPCML, RNASE1 and YES1） was similar to tumor suppressor genes. For example, the expression level of these genes was the highest in normal gastric epithelium, which was decreased in primary carcinoma, and further decreased in metastatic lymph nodes. On the contrary, the expression pattern of gene ACK1 was similar to that of oncogene. Four genes were further identified as differentially expressed genes in the majority of the cases in the progression of gastric cancer. CONCLUSION： LMD in combination with cDNA microaro ray provides a unique support foe the identification of early expression profiles of differential genes and the expression pattern of 3 genes （OPCML, RNASE1 and YES1） associated with the progression of gastric cancer. Further study is needed to reveal the molecular mechanism of lymph node metastasis in patients with gastric cancer.

Three new alternative splicing variants of human cytochrome P450 2D6 mRNA in human extratumoral liver tissue

AIM: To identify the new alternative splicing variants of human CYP2D6 in human extratumoral liver tissue with RT-PCR and sequencing. METHODS: Full length of human CYP2D6 cDNAs was amplificated by reverse transcription-polymerase chain reaction (RT-PCR) from a human extratumoral liver tissue and cloned into pGEM-T vector. The cDNA was sequenced. Exons from 1 to 4 of human CYP2D6 cDNAs were also amplificated by RT-PCR from extratumoral liver tissues of 17 human hepatocellular carcinomas. Some RT-PCR products were sequenced. Exons 1 to 4 of CYP2D6 gene were amplified by PCR from extratumoral liver tissue DNA. Two PCR products from extratumoral liver tissues expressing skipped mRNA were partially sequenced. RESULTS: One of the CYP2D6 cDNAs had 470 nucleotides from 79 to 548 (3' portion of exons 1 to 5' portion of exon 4), and was skipped. Exons 1 to 4 of CYP2D6 cDNA were assayed with RT-PCR in 17 extratumoral liver tissues. Both wild type and skipped mRNAs were expressed in 4 samples, only wild type mRNA was expressed in 5 samples, and only skipped mRNA was expressed in 8 samples. Two more variants were identified by sequencing the RT-PCR products of exons 1 to 4 of CYP2D6 cDNA. The second variant skipped 411 nucleotides from 175 to 585. This variant was identified in 4 different liver tissues by sequencing the RT-PCR products. We sequenced partially 2 of the PCR products amplified of CYP2D6 exon 1 to exon 4 from extratumoral liver tissue genomic DNA that only expressed skipped mRNA by RT-PCR. No point mutations around exon 1, intron 1, and exon 4, and no deletion in CYP2D6 gene were detected. The third variant was the skipped exon 3, and 153 bp was lost. CONCLUSION: Three new alternative splicing variants of CYP2D6 mRNA have been identified. They may not be caused by gene mutation and may lose CYP2D6 activity and act as a down-regulator of CYP2D6.

Comparison of the Tellgenplex HPV DNA test with the PCR-reverse dot blot assay for human papillomavirus genotyping

Objective: To access the performance of the Tellgenplex human papillomavirus(HPV) DNA test compared to the polymerase chain reaction-reverse dot blot(PCR-RDB) assay for the HPV genotyping.Methods: Sixty cervical swab samples were genotyped by the Tellgenplex HPV DNA test and the PCR-RDB assay.The Tellgenplex HPV DNA test and the PCR-RDB assay can detect 26 and 23 HPV genotypes, respectively.Each sample showed discrepancy was genotyped using sequencing.Results: The percent agreement between the two tests ranged from 83.3% to 100.0% according to different genotype.This showed perfect agreement(>0.81) for high-risk HPV genotypes(35, 39, 45, 53, 56, 59, 66, 68, and 82), substantial agreement(>0.65) for high-risk HPV genotypes(16, 18, 33, 52, and 58) and low-risk HPV genotype 43 between the two assays by the kappa analysis.The positive rates of the two assays for frequent HPV genotypes(16, 35, 39, 45, 52, 53, 58, 59, 66, and 82) were not statistically different, but the PCR-RDB assay showed higher positive rates than the Tellgenplex HPV DNA test for HPV genotypes 81(P<0.05).As for more than 10 positive results by the Tellgenplex HPV DNA test and/or the PCR-RDB assay, the PCR-RDB assay showed higher relative sensitivity and specificity than the Tellgenplex HPV DNA test for the three HPV genotypes(16, 52, and 81).All HPV genotypes that can be detected by only the Tellgenplex HPV DNA test(HPV genotypes 44 and 55) were confirmed by sequencing.Conclusions: In conclusion, our results demonstrated that the PCR-RDB assay which can detect more multiple HPV genotypes in each specimen shows higher relative sensitivity and specificity than the Tellgenplex HPV DNA test, which makes it a better option for routine clinical use.

Expression of tumor necrosis factor related apoptosis inducing ligand receptor in glioblastoma

BACKGROUND： Receptors for tumor necrosis factor related apoptosis inducing ligand （TRAIL） include death receptor 4, death receptor 5, decoy receptor 1, and decoy receptor 2. Activation of death receptor 4 and 5 selectively kills tumor cells. OBJECTIVE： To detect TRAIL receptor expression in glioblastoma by immunohistochemistry and RT-PCR, and to compare this expression to that in normal brain tissue. DESIGN： Observational analysis. SETTING： Department of Pathology, the First Affiliated Hospital of Zhengzhou University; Henan Tumor Pathology Key Laboratory. PARTICIPANTS： Twenty-five patients （17 males and 8 females） who received glioblastoma resection were selected from the Fifth Affiliated Hospital of Zhengzhou University, between September 2003 to June 2004. All glioblastoma samples were diagnosed pathologically. Twenty patients （12 males and 8 females） with craniocerebral injury who received normal brain tissue resection were selected in the same time period. There were no significant differences in sex and age between glioblastoma patients or between craniocerebral injury patients （P 〉 0.05）. All patients and appropriate relatives provided informed consent, and this study was approved by the local research ethics committee. METHODS： Polyclonal antibody against TRAIL receptors and an immunohistochemical kit （batch number： 200502） were purchased from Boster Company, Wuhan. Immunohistochemistry： Expression of death receptor 4, death receptor 5, decoy receptor l, and decoy receptor 2 were observed in both glioblastoma and normal brain tissue. The experiment was performed according to the kit instructions, and positive staining was brown-yellow. Assessment： There were no positive signals （-）; weakly positive signals, positive cells 〈 25% （＋）; weakly positive signals, positive cells 25%-50% （＋＋）; strongly positive signals, positive cells 50%-75% （＋＋＋）; strongly positive signals, positive cells 〉 75% （＋＋＋＋）. Evaluation： Expression levels of TRAIL receptors were estimated in both normal brain tissue and glioblastoma. Expression of decoy receptor 1 and decoy receptor 2 mRNA in glioblastoma were detected by reverse transcription polymerase chain reaction, and expression of decoy receptor in glioblastoma was estimated. MAIN OUTCOME MEASURES： Comparison of death receptor and decoy receptor protein expression between glioblastoma and normal brain tissue; decoy receptor mRNA expression in glioblastoma. RESULTS： Death receptor protein expression was strongly positive （＋＋＋） in glioblastoma, while it was weakly positive （＋, ＋＋） in normal brain tissue. Therefore, expression rate of death receptor protein in the glioblastoma was significantly higher than that in the normal brain tissue （.~ 2 = 18.48, 23.03, P 〈 0.01）. Decoy receptor protein expression in the glioblastoma was significantly lower than that in the normal brain tissue （ x2 = 6.65, 18.76, P 〈 0.01）. The level of decoy receptor mRNA expression in glioblastoma was significantly higher than those of protein expression （ x 2 = 9.82, 10.09, P〈 0.01）. CONCLUSION： High expression of death receptor and low expression of decoy receptor are frequently observed in glioblastoma, suggesting that TRAIL receptor genes show an anti-tumor and expressive response during the initiation and development of the tumor. There are significant differences in decoy receptor expression between normal brain tissue and glioblastoma, suggesting that the restricted expression of decoy receptor in glioblastoma is regulated at the post-transcriptional level.

A rapid reverse dot blot assay for all 18 β-thalassemia mutations in Chinese population

A set of allele-specific oligonucleotide (ASO) probes used for detecting all 18 β-tha-lassemia mutations found in Chinese was immobilized on two strips of Biodyne C membrane;one containing 7 pairs of oligonucleotide probes specific for the most commonly found mutant al-leles,and the other containing the remaining 11 pairs of ASO_s specific for the less commonlyfound.The membranes were hybridized with β-globin sequences amplified by polymerase chainreaction (PCR) with biotinylated primers,and then treated with Streptavidin-POD conjugateand substrates for color development.The method has been applied successfully to the detectionof all 18 Chinese β-thalassemia mutations and prenatal diagnosis of two high-risk pregnancies ofβ-thalassemia.Patients with homozygous,heterozygous and compound heterozygous alleles ofthese mutations and normal individuals could be easily distinguished by the present method.Us-ing the immobilized-probe format (reverse dot blot),it was able to screen simultaneously multi-ple β-thalassemia mutations of a DNA sample by performing hybridization only once.This assayis simple,rapid and independent of radio-isotopes and can be appplied for all 18 β-thalassemiamutations so far found in Chinese population.It is considered that this method may be usefulfor gene frequency investigation of large numbers of β-thalassemia DNA samples and used as aroutine method in the clinic laboratory.

Quantification of mRNA Levels by Fluorescently Labelled Reverse Transcription Competitive PCR

A reproducible,quantitative,non-radioactive method for the analysis of mRNA expression is described.After RNA preparation and cDNA synthesis,the cDNA was co-amplified with an internal standard in the same PCR system.The PCR products containing both targen and internal standard amplificates were electrophoresed and detected on an ABI 377 DNA Sequencer.For each sample,β-actin was also quantified by an identical procedure to compensate for relative differences between samples in the integrity of the individual RNA samples and for variations in reverse transcription.Due to the linear relationship between cDNA content and PCR product ratio of target cDNA template and competitive standard,a single PCR reaction was sufficient for quantification of a sample.The experimental results showed that the method is a mRNA quantitative RT-PCR method with high sensitivity and good reproducibility.It can be used in large-scale accurate quantitative analyses of mRNA expression of any gene.

肝癌患者血清中lncRNA HOTAIR表达与临床病理特征的相关性及诊断价值

目的分析血清长链非编码RNA HOX转录反义RNA(IncRNA HOTAIR)在肝癌患者中的表达与临床诊断意义。方法选取100例肝癌患者为研究对象,并将其纳入肝癌组,另选取同期收治的80例肝硬化患者纳入肝硬化组,同时选取同期行体检的75例健康体检者纳入对照组。采集3组5 ml静脉血,以实时荧光定量逆转录聚合酶链反应(RT-PCR)检测对比3组血清IncRNA HOTAIR相对表达量。同时,收集肝癌患者的年龄、性别等一般资料,分析血清IncRNA HOTAIR与其临床病理特征的关系;另外绘制受试者工作曲线(ROC),分析血清IncRNA HOTAIR在肝癌中的诊断效能。结果肝癌组的IncRNA HOTAIR相对表达量[(2.53±0.58)]高于肝硬化组[(0.81±0.25)]与对照组[(0.20±0.05)],差异有统计学意义(P<0.05)。IncRNA HOTAIR表达与肝癌患者的TNM分期、血管侵犯、肝外转移有关,差异有统计学意义(P<0.05);IncRNA HOTAIR表达与患者的年龄、性别、肿瘤数目无关,差异无统计学意义(P>0.05)。ROC结果显示:血清IncRNA HOTAIR诊断肝癌的AUC为0.901(95%CI:0.852~0.949),具有较高的诊断价值。结论肝癌患者血清内IncRNA HOTAIR呈高表达,其表达水平与患者TNM分期、血管侵犯、肝外转移相关,可能参与肝癌的发生、发展,在肝癌中具有较高的临床诊断意义。

2019—2021年安徽省合肥市某三甲医院住院的严重急性呼吸道感染患者的监测分析

目的分析2019—2021年在安徽省合肥市第二人民医院住院的严重急性呼吸道感染(SARI)患者的监测结果。方法筛查2019年1月—2021年12月安徽省合肥市第二人民医院住院部监测科室(呼吸内科、儿内科与重症监护科)SARI患者的资料,并采集患者鼻咽拭子、呼吸道分泌物,通过反转录聚合酶链反应(RT-PCR)检测甲乙通用性流感病毒。结果2019—2021年,共监测SARI患者934例,占监测科室同期入院病例的12.04%;2019年、2020年和2021年,SARI患者分别占入院患者的10.62%、14.90%和11.40%。2020年,检测出的SARI患者占比较高。2019—2021年,SARI监测病例主要集中于冬春季节,以5岁以下儿童病例占比较大(54.82%),男女比例为1.66∶1。SARI的监测科室分别为儿内科(82.12%)、呼吸内科(17.77%)以及重症监护科(0.11%);2019—2021年采集的934份咽拭子标本经实验室检测,阳性标本39份,阳性率为4.17%,主要感染病毒类型为B型Victoria系[27例(69.23%)]。SARI与流感阳性感染患者中合并症最常见的为肺炎,SARI合并并发症的467例患者中,99.36%的患者转归为治愈或好转,不合并并发症的患者中,99.57%的患者转归为治愈或好转,差异无统计学意义(P=1.000)。SARI病例采样前最常用的药物为抗生素与神经氨酸酶抑制剂类抗病毒药物。SARI病例中,使用抗病毒药物的患者中有98.66%疾病转归为治愈或好转,而不使用抗病毒药物的患者中,99.53%的患者疾病转归为治愈或好转,差异无统计学意义(P=0.343)。结论2019—2021年,在安徽省合肥市第二人民医院住院的SARI病例多为男性、儿童、老人,冬春季为高发季节,流感阳性患者主要感染菌株均为FluB,且所有患者转归情况较好。

急性胰腺炎病人血清长链非编码RNA核富集转录体1、Toll样受体2表达变化及临床意义研究

目的探讨急性胰腺炎(AP)病人血清长链非编码RNA核富集转录体1(LncRNA NETA1)、Toll样受体2(TLR2)表达及临床意义。方法选取2017年5月至2020年12月石家庄平安医院收治的AP病人125例为研究对象,根据急性生理学和慢性健康状况评价Ⅱ(APACHEⅡ)对AP病人进行评分,并将病人分为轻症组56例,重症组69例;根据住院期间重症组AP病人预后情况,将病人分为预后不良25例,预后良好44例。同期选取该院健康体检者130例为健康组。实时荧光定量逆转录聚合酶链反应(qRT-PCR)检测血清LncRNA NETA1水平,酶联免疫吸附测定(ELISA)检测血清TLR2水平;Pearson相关性分析探索AP病人血清LncRNA NEAT1、TLR2水平相关性及二者与病人APACHEⅡ评分的关系;受试者操作特征曲线(ROC曲线)分析血清LncRNA NEAT1、TLR2对AP病人重症的评估价值。结果与健康组比较,AP组血清LncRNA NEAT1(2.16±0.31比1.00±0.00)、TLR2水平[(6.43±1.05)µg/L比(0.59±0.24)µg/L]升高(P<0.05);与轻症组比较,重症组AP病人血清LncRNA NEAT1(2.31±0.33比1.98±0.27)、TLR2水平[(6.89±1.16)µg/L比(5.86±0.92)µg/L]升高(P<0.05);与预后良好组比较,预后不良组重症AP病人血清LncRNA NEAT1(2.43±0.36比2.14±0.27)、TLR2水平[(7.12±1.24)µg/L比(6.45±1.02)µg/L]升高(P<0.05);Pearson相关性分析结果表明,AP病人血清LncRNA NEAT1、TLR2呈正相关(r=0.65,P<0.05),二者与病人APACHEⅡ评分均呈正相关(r=0.50、0.52,P<0.05);ROC曲线结果显示,血清LncRNA NEAT1水平、TLR2单独及联合评估AP病人重症的曲线下面积(AUC)分别为0.73[95%CI:(0.64,0.82)]、0.75[95%CI:(0.67,0.84)]、0.88[95%CI:(0.82,0.94)],血清LncRNA NEAT1联合TLR2水平预测AP病人重症的AUC明显大于二者单独预测(P<0.05)。结论AP病人血清LncRNA NEAT1、TLR2表达水平显著升高,且与病人严重程度和预后有关,临床上检测LncRNA NEAT1、TLR2表达可能有助于AP病人的病情评估。

常见白血病融合基因筛查在白血病诊断与分型中的意义

目的: 分析病人白血病细胞染色体畸变涉及的86种融合基因和临床白血病类型的相关性,探讨常见融合基因筛查法在临床诊断和分型中的应用价值。方法: 收集161例初发或者复发的白血病患者及8例骨髓增生异常综合征(MDS)患者的骨髓细胞,提取RNA,用32条特异性引物逆转录为cDNA,利用白血病29种染色体畸变形成的融合基因的86种mRNA剪接变异体引物,分8管进行多重RT PCR,筛查白血病融合基因。结合临床状态和形态学观察了解融合基因与白血病类型的关系。结果: 白血病中115例(71% )分别检测出10种白血病常见融合基因,包括AML1 /ETO、PML/RARα、PLZF/RARα、dupMLL、MLL/AF6、MLL/AF10、CBFβ/MYH11、BCR/ABL、Hox11、Evi1。其中52例慢性粒细胞白血病(CML)100%检出BCR/ABL; 25例急性早幼粒细胞白血病(APL)中88%检出融合基因,其中21例APL检测出PML/RARα, 1例APL检测出PLZF/RARα; AML1 /ETO阳性的17例急性白血病(AL)16例为FAB M2亚型, 1例为混合型白血病; CBFβ/MYH11阳性的4例AL3例为FAB分型的M4, 1例为M5,属于向粒单细胞系统分化的白血病。16例AL检测出MLL基因异常,其中MLL/AF6白血病均为FAB分型的M5,具有典型的原始单核细胞白血病的特征。17例急性淋巴细胞白血病(ALL) 5例检测出BCR/ABL。8例MDS病人中2例检测出融?

中药滋补肝肾方对鼠B16黑素瘤细胞酪氨酸酶mRNA表达的调节

目的 探讨中药滋补肝肾方治疗白癜风的分子学作用机制。方法 采用定量逆转录 聚合酶链式反应(RT PCR)方法 ,观察中药滋补肝肾方对鼠B16黑素瘤细胞株酪氨酸酶mRNA表达的影响。结果 中药滋补肝肾方能使黑素瘤细胞酪氨酸酶mRNA表达水平增加。结论 中药滋补肝肾方能上调黑素细胞酪氨酸酶mRNA表达水平 ,这是其治疗白癜风的药理作用机制之一。

大鼠关节软骨酸敏感离子通道的表达

目的探讨大鼠关节软骨酸敏感离子通道(ASICs)的表达及其意义。方法利用RT-PCR和Western blot检测ASICs在大鼠关节软骨的表达。结果大鼠关节软骨存在ASIC1a、ASIC2a和ASIC3 mRNA及其蛋白的表达。结论ASICs在大鼠关节软骨组织的表达,其有望成为关节炎治疗的新靶点。

大豆异黄酮对胰岛素抵抗大鼠胰岛素敏感性及瘦素基因表达的影响

目的:观察大豆异黄酮(SIF)对高脂膳食诱导的胰岛素抵抗(IR)大鼠血清瘦素水平及白色脂肪组织瘦素mRNA表达的影响,并探讨SIF提高胰岛素敏感性的可能机制。方法:选用高脂饲料诱导的IR雄性SD大鼠,根据其胰岛素抵抗指数(IRI)随机分为模型对照组(水)和3个SIF组(50 mg／kg,150 mg/kg及450 mg／kg)。各组经口灌胃给予相应受试物1个月,空腹过夜后取材,用酶法检测各组动物空腹血糖、用放射免疫法检测空腹血胰岛素、用酶联免疫法检测血清瘦素含量,以实时定量RT-PCR法检测肾周脂肪组织瘦素基因的mRNA水平。结果:与模型对照组比较,SIF 150 mg／kg和450 mg／kg组能显著降低空腹血胰岛素水平及IRI,SIF450 mg／kg组能明显降低大鼠体重及肾周脂肪组织瘦素基因mRNA的表达、提高血清瘦素的含量。结论:SIF可能具有改善瘦素抵抗的作用,并进而提高胰岛素的敏感性。