Study on PVL, blaOXA-23 and blaOXA-51 Genes in Drug Resistant Staphylococcus aureus Causing Surgical-Sites and Traumatic Wounds Infections, Sudan

Background: The characteristics of Staphylococcus aureus that made it the most important cause of wound infections are environmental spread antimicrobials resistance and virulence. Absence of molecular detection of drug resistance and virulence factors in many developing countries limits the epidemiological information. This study conducted to identify PVL virulence gene, and blaOXA-23 and blaOXA-51 drug resistance genes of Staphylococcus aureus isolated from surgical-sites infections (SSIs) and traumatic wounds. Methods: A cross-sectional study was conducted from 2019 to 2021, in which 70 cefepime resistant Staphylococcus aureus were used, the strains were isolated from patients of SSIs and traumatic wounds admitted to the department of General Surgery in Wad Medani Teaching Hospital. Mannitol salt agar was used for primary culture followed by biochemical identification and Kirby Bauer susceptibility testing. Single and multiplex PCR protocols performed for bacterial confirmation and target genes detection. Results: Staphylococcus aureus strains from SSIs constituted 56% (39/70) from which 41% (16/39) possessed PVL gene while 42% (13/31) of wound infections strains were positive for PVL gene. Presence of PVL gene was significantly associated with resistance to meropenem (P. value 0.023) and ceftriaxone (P. value 0.037). blaOXA-23 was significantly detected with resistance to meropenem, augmentin and ceftriaxone. While blaOXA-51 was significantly identified among Staphylococcus aureus strains that showed resistance to meropenem and ciprofloxacin. Conclusion: This is the first study in Sudan that identified blaOXA-23 and blaOXA-51 in Staphylococcus aureus and correlated them to resistance to commonly used antimicrobials. Meropenem resistant Staphylococcus aureus were significantly positive for PVL, blaOXA-23 and baOXA-51 genes.

Cloning and Bioinformatics Analysis of P23 Gene from Theileria sergenti

[Objective] The aim of this study is to provide basis for developing genetic engineering vaccine and diagnostic kit for Theileria sergenti infection. [Objective] P23 gene of Theileria sergenti was amplified from its genomic DNA by PCR amplification, and cloned into the pGEM-Easy vector; then the sequencing result was analyzed with bioinformatics methods. [Result] Whole length of the P23 gene from Theileria sergenti is 684 bp containing a 672 bp open reading frame. The deduced amino acid sequence (223 amino acid residues) contains a signal peptide of 19 amino acid residues and two fragments of transmembrane domains, with relative molecular weight of the 25.886 kD and with the pI of 9.22. The homology between the yielded sequence and Chitose of Theileria sergenti P23 gene(TS-Chitose type, D84446), Ikeda of Theileria sergenti P23 gene(TS-Ikeda type, D84447) reached 99% and 90%, respectively. The sequence has been accessed in GenBank(EU573168). [Conclusion] The protein encoded by the P23 gene has better stability and immunogenicity, thus can be used as the antigen candidate for preparing genetic engineering vaccine for Theileria sergenti.

Cloning and Prokaryotic Expression of P23 Major Surface Protein Gene from Theileria sergenti

[Objective] The aim was to study cloning and prokaryotic expression of P23 major surface protein gene of Theileria sergenti. [Method] A pair of specific primers was designed according to the sequence of P23 major surface protein of T. sergenti (D84447).The P23 gene was amplified by PCR from genomic DNA of T. sergenti and cloned into pMD18-T vector to construct recombinant clonal vector pMD18-P23. Positive clones were identified by PCR screening and restriction digestion. A recombinant expression plasmid pGEX-4T-P23 was constructed by subcloning the cloned P23 gene into the linearized pGEX-4T-1 vector and transformed into E. coli BL21. After introduction by IPTG,the expressed fusion protein was identified by SDS-PAGE and Western-blotting. [Result] The cloned gene has a total length of 507 bp. Sequencing result showed that the nucleotide sequence of the cloned P23 gene shared 99.4% identity with that of P23 published in GenBank (D84447). The expressed fusion protein was 46 ku in molecular mass. Induction opportunity of zhours after culture inoculation was the best,the induction time of 6 h was the best,and induction temperature of 34 ℃ was the best as well,IPTG of 1 mmol/L had little effect on the expression. Western-blotting indicated that recombinant protein was recognized by specific antibody. [Conclusion] This study would lay a foundation for further research on the prevention and diagnose of T. sergenti.

盐亭县人群白细胞介素-23受体基因多态性与上尿路结石易感性的关联

目的:研究盐亭县人群白细胞介素-23(IL-23)受体基因多态性与上尿路结石易感性的关联。方法:应用简单随机抽样法选取盐亭县人民医院2020年2月—2023年2月收治的上尿路结石患者104例作为结石组,另选取100例健康体检者为健康组。比较两组IL-23受体基因相关位点的多态性,分析IL-23受体基因多态性与上尿路结石易感性的关系。结果:结石组IL-23受体基因rs10889677位点AA基因型人数占比低于健康组,CC基因型人数占比高于健康组,差异有统计学意义(P<0.05)。结石组IL-23受体基因rs11465817位点AA基因型人数占比高于健康组,CC基因型人数占比低于健康组,差异有统计学意义(P<0.05)。IL-23受体基因rs10889677位点基因型为CC型、IL-23受体基因rs11465817位点基因型为AA型是盐亭县人群上尿路结石易感性的独立危险因素(P<0.001)。结论:IL-23受体基因rs10889677位点基因型为CC型、IL-23受体基因rs11465817位点基因型为AA型是盐亭县人群上尿路结石易感性的独立危险因素,提示IL-23受体基因多态性的检测可为早期识别上尿路结石易感人群及后续预防性干预措施的制定和实施提供参考。

Expression of nm23 gene in hepatocellular carcinoma tissue and its relation with metastasis

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The 5-HT2c receptor gene Cys23Ser polymorphism influences the intravaginal ejaculation latency time in Dutch Caucasian men with lifelong premature ejaculation

It has been postulated that the persistent short intravaginal ejaculation latency time （IELT） of men with lifelong premature ejaculation （LPE） is related to 5-hydroxytryptamine （HT）2c receptor functioning. The aim of this study was to investigate the relationship of Cys23Ser 5-HT2c receptor gene polymorphism and the duration of IELT in men with LPE. Therefore, a prospective study was conducted in 64 Dutch Caucasian men with LPE. Baseline IELT during coitus was assessed by stopwatch over a 1-month period. All men were genotyped for Cys23Ser 5-HT2c receptor gene polymorphism. Allele frequencies and genotypes of Cys and Ser variants of 5-HT2c receptor gene polymorphism were determined. Association between Cys/Cys and Ser/Ser genotypes and the natural logarithm of the IELT in men with LPE were.investigated. As a result, the geometric mean, median and natural mean IELT were 25.2, 27.0, 33.9s, respectively. Of all men, 20.0%, 10.8%, 23.1% and 41.5% ejaculated within 10, 10-20, 20-30 and 30-60s after vaginal penetration. Of the 64 men, the Cys/Cys and Ser/Ser genotype frequency for the Cys23Ser polymorphism of the 5-HT2c receptor gene was 81% and 19%, respectively. The geometric mean IELT of the wildtypes （Cys/Cys） is significantly lower （22.6s; 95% CI 18.3-27.8s） than in male homozygous mutants （Ser/Ser） （40.4s; 95% CI 20.3-80.4s） （P = 0.03）. It is concluded that Cys23Ser 5-HT2c receptor gene polymorphism is associated with the IELT in men with LPE. Men with Cys/Cys genotype have shorter IELTs than men with Ser/Ser genotypes.

NORTHERN BLOT ANALYSIS OF nm23 GENE EXPRESSION IN HUMAN LUNG CANCER

Objective: To investigate the role of nm23 gene expression in human lung cancer. Methods: Forty human lung cancer tissues and 19 non-cancer pulmonary tissues were studied for their nm23-H1 and nm23-H2 mRNA expression with non-radioactive Northern blot hybridization. The correlation of nm23 mRNA expression with clinical features of lung cancer was analyzed. Results: The mRNA expression of nm23-H2 gene in poorly differentiated squamous cell carcinoma was significantly decreased compared to that in moderate-high differentiated squamousd cell carcinoma. The mRNA expression of nm23-H1 and nm23-H2 gene in small cell lung cancer was significantly decreased compared to that in squamous cell carcinoma. No significant difference in nm23 mRNA expression was observed between lung cancer with and without lymph node metastasis, nor was there significant difference between tumor stage. Conclusion: The mRNA expression of nm23 gene is correlated with the degree of differentiation of lung cancer, but there is no evidence of metastasis suppression effect by nm23 gene.

EVALUATION OF THE PROGNOSTIC VALUE OF nm23 GENE EXPRESSION IN BREAST CANCER

Objective: To investigate the expression of nm23 gene and evaluate its prognostic value in breast cancer. Methods: nm23 expressions were detected in 101 breast cancer patients (group 1) by immunohistochemistry. RT-PCR and immunohistochemistry were used to measure expressions of nm23 gene in another 68 patients with breast cancer (group 2). Results: nm23 gene expression in group 1 was inversely associated with distant metastasis and lymph node metastasis (P<0.05). In 44 patients with negative lymph node, 9 cases progressed to distant metastasis, 7 of them (77.8%) showed low expression of nm23 gene (P<0.05). In 57 patients with positive lymph node, 24 our of 29 patients who had no distant metastasis (82.8%) expressed nm23 gene at high level (P<0.05). Meanwhile, there were 6 patients with distant metastasis in the group 2, all of thenm expressed nm23 gene mRNA at low level. Conclusion: The results showed that nm23 gene might play an independent role in predicting prognosis of breast cancer.

Incidence of Oxa23 and Oxa51 Genes Associated with Bacterial Isolated from Patients with Urosepsis: Single Centre Prespective

Background: Urosepsis is one of the most common infections that require empirical broad spectrum antibiotics immediately after diagnosis. This has led to development of bacterial resistance by acquiring the capability to destroy the β-lactam ring. Methodology: This is a cross-sectional hospital-based study. The study was conducted from 2019 to 2020 at Gezira Hospital for Renal diseases and surgery (GHRDS). A hundred patients were diagnosed clinically with urosepsis and the isolated organisms were Escherichia coli, Staphylococcus aureus, Proteus mirabilis, Klebsiella pneumonia and Pseudomonas aeruginosa. The susceptibility test was conducted by Kirby Bauer disc diffusion technique according to clinical laboratory standard institute (CLSI) guidelines. Seventy eight samples of bacterial genomic DNA were confirmed by 16srRNA and multiplex PCR, were performed for genotypic blaOXA-51 and blaOXA-23 gene characterization of isolated bacteria. Then gel electrophoresis was used to identify the presence or absence of (blaOXA-51 and blaOXA-23) genes. Results: 88.5% (69/78) in 16srRNA detected. Using multiplex PCR, the frequencies of blaOXA-51 and blaOXA-23 genes were 13% and 10.1%, respectively. The percentages of isolates which yielded both blaOXA-51 and blaOXA-23 among P. aeruginosa was 25% (1/4), among K. pneumonia was 17% (1/6), and among E. coli was 8% (3/37). Only blaOXA-51 was detected in P. mirabilis 10% (1/10) and only blaOXA-23 was detected in S. aureus 5% (1/18). Conclusion: In this study, the presence of blaOXA-51 and blaOXA-23 genes was increased in the isolated bacteria.

EXPRESSION OF NM23-H1 GENE PRODUCT IN NASOPHARYNGEAL CARCINOMA AND ITS CLINICAL SIGNIFICANCE

Objective: The nm23 gene is one of the tumor metastatic suppressor genes. The expression of nm23H1 has been reported to be inversely associated with metastatic potentiality in a number of human carcinomas, including breast, colorectal, gastric, hepatocellular and gallbladder carcinomas. In this study, the immunohistochemical staining of nm23H1 protein in human nasopharyngeal carcinoma (NPC) was examined, and the relationship between nm23H1 and both metastasis and prognosis of patients with NPC was also investigated. Methods: Routine LSAB immunohistochemistry with the nm23H1 monoclonal murine antibody was employed to study the expression of nm23H1 protein in 95 paraffinembedded specimens of NPC treated at our hospital. The clinical pathologic data and results of followup were also retrieved. Comparisons between patients with and without expression of nm23H1 protein with respect to metastasis, locoregional recurrence and survival were performed using Log rank test. Multivariate prognostic analyses were performed by using Cox's regression model. Results: Nm23H1 negative expressive tumors were associated with a higher incidence of lymphnode metastasis (86.7%) than those of nm23H1 positive (48.6%, P<0.01). Nm23H1 negative expressive tumors were associated with a high incidence of recurrence and distant metastasis after radiotherapy (P<0.05). A significant association was found between expression of nm23H1 and prognosis (P<0.01). The expression of nm23H1 indicated favorable prognosis. Conclusion: It was suggested that nm23 H1 negative expression was significantly associated with lymphnode metastasis, recurrence and distant metastasis. Nm23H1 may have value for predicting the prognosis of NPC.

THE HIGH-LEVEL EXPRESSION OF nm23(NDP)GENE IN NPC

We have studied the expression of nm23(NDP) in 50 cases nasopharyngeal biopsies with anti-nm23(NDP) antibodies. As a result, the NDP positive rate in nasopharyngeal carcinoma (NPC) (95.54%) markedly increased (P<0.05), as compared with that in the normal nasopharyngeal epithelia (50.00-60.00%) and lymphocytes (52.00%). There were cytopfasmic type, nucleus type and mixed cytoplasmonucleus type according to NDP location in a cell. Their positive rates were 64.44%, 15.56% and 20.00% respectively in nasopharyngeal carcinoma. The expression of NDP had no relation with cervical lymphometastases in NPC, and the NDP positive rates had no significance between bilateral cervical lymphometastases and unilateral (P<0.05). But the NDP expression had most relation with the NPC staging. The expression rate and the intensity in Ⅲ or Ⅳ stage patients were markedly higher than that in II stage. It points out that the high-level expression of NDP had relation with the rapid cellular proliferation in NPC, and it may indicate the bad prognoses.

Expression of the novel gene NM23-H1B in ovarian cancer

Objective:To study the expression of the human novel gene NM23-H1B in ovarian cancer. Methods: Totally 24 samples from patients with epithelial ovarian tumor at different clinical stages and 4 from normal ovaries were examined for NM23-H1B mRNA expression by RT-PCR and Northern blot. Results: All samples expressed NM23-H1B mRNA through RT-PCR, while the level of expression in ovarian tumor was higher than that of normal ovary. The results of Northern blot showed that NM23-H1B was overexpressed in ovarian cancer while lowexpressed in normal ovary or low malignant potential (LMP). The level of expression at early stage cancer(stageⅠand Ⅱ) was higher than those in advanced cancer(stage Ⅲ and Ⅳ). In early stage carcinoma, the expression level was involved in the differentiation of tumor cell, and well-differentiated cancer expressed NM23-H1B mRNA in comparatively higher level. Conclusion: The novel gene NM23-N1B is closely correlated with the ovarian cancer.

IDENTIFICATION OF A NEW HUMAN nm23 GENE,nm23-H3b

Nm23 is a kind of effective tumor metastasis suppressor genes which included two types in human:nm23-H1 and nm23-H2. Amino acid identity between nm23-H1 and nm23-H2 was 88%.In this study,using a pair of primers to flank the part of coding sequence of nm 23,the 5'-translated sequence was amplified by polymerase chain reaction (PCR) from human normal liver genomic DNA.A 375-base pairs clone was charactertzed,which designated pnm23-H3b.The nm23-H3b nucleotide sequence between 40 bp and 70 bp is different from nm23-H1 and nm23-H2,and other sequences have 86%and 90%identical to nm23-H1 and nm23-H2,respectively.Southern blot containing BglⅡ-digested human liver genomic DNA hybridized to the entire nm23-H3b DNA and showed three bands at 10.5,7.9 and 4.0 Kb.These data demonstrate that the third human nm23 exists possibly.Therefore,nm23 may be considered a family of closely related genes.

Relationship between IL-23R rs11209026 Gene Polymorphism and Susceptibility to Acute Coronary Syndrome

Background: Several studies illustrated that IL-23/Th17 axis could be an important pro-inflammatory pathway in atherosclerosis. As a key element in this inflammatory mechanism, interleukin-23 receptor (IL-23R) may play a critical role in the pathological process of atherosclerosis. Single nucleotide polymorphisms (SNPs) in IL-23R have recently been consistently found to be associated with atherosclerosis diseases. However, its association with acute coronary syndrome (ACS) is still indistinct. Here, we discussed whether genetic polymorphisms in IL-23R (rs11209026 G/A) were associated with susceptibility to ACS. Methods: Among 160 patients with ACS, it includes 80 patients with unstable angina pectoris (UAP), 80 patients with myocardial infarction (MI), and 80 control subjects were selected randomly. The polymorphisms of IL-23R (rs11209026 G/A) were analyzed by the Sanger method. Results: Data showed that percentages of rs11209026 AG genotypes were significantly lower in ACS group (including: UAP and MI) than in controls (odds ratio [OR] = 0.324, 95% confidence interval [CI]: 0.148 - 0.712, p = 0.005;OR = 0.351, 95% CI: 0.135 - 0.910, p = 0.031;OR = 0.303, 95% CI: 0.112 - 0.817, p = 0.018, respectively). Furthermore, there was no correlation between rs11209026 G/A SNP and dyslipidemia-associated ACS. Conclusions: The variant of the rs11209026 polymorphism in IL-23R gene might decrease the risk of ACS, and these data suggest that AG genotype of the rs11209026 G/A polymorphism may act as a protective factor for acute coronary syndrome and subtype of ACS.

血清成纤维生长因子-23/抗衰老基因Klotho、辅助性T淋巴细胞1/辅助性T淋巴细胞2细胞因子在老年终末期肾病患者中的表达及对医院感染的预测和预后的影响

目的观察血清成纤维生长因子-23(fibroblast growth factor 23,FGF23)/抗衰老基因Klotho、辅助性T淋巴细胞(helper T lymphocytes,Th)1/Th2细胞因子在老年终末期肾病患者中的表达,并分析对医院感染的预测和预后的影响。方法选取2021年1月1日至2022年12月31日自贡市精神卫生中心(自贡市老年病医院)收治的106例老年终末期肾病患者,男65例,年龄范围60~84岁,年龄(67.21±3.07)岁,女41例,年龄范围60~84岁,年龄(67.65±2.98)岁,于入院第2天检测血清FGF23、抗衰老基因Klotho、Th1细胞因子[γ干扰素(interferon-γ,IFN-γ)、白介素2(interleukin-2,IL-2)]、Th2细胞因子(IL-4、IL-10)水平,进行急性生理功能及慢性健康状况评分Ⅱ(acute physiology and chronic health status scoring systemⅡ,APACHEⅡ)评估。根据住院期间是否发生医院感染分为感染组(28例)、未感染组(78例),比较两组各项血清指标及APACHEⅡ评分水平,分析各项指标与APACHEⅡ评分的相关性。随访3个月统计患者生存预后[生存(71例)、死亡(35例)],比较不同预后患者血清FGF23、Klotho、IFN-γ、IL-2、IL-4、IL-10水平,分析各项指标对医院感染及预后的预测价值及临床效用。结果106例老年终末期肾病患者根据住院期间是否发生医院感染分为感染组(28例)、未感染组(78例),随访3个月后生存71例、死亡35例。入院第2天感染组血清FGF23、IL-4、IL-10水平及APACHEⅡ评分分别为(78.64±20.16)ng/mL、(20.14±1.48)μg/L、(22.47±2.56)μg/L、(26.38±6.51)分,未感染组分别为(60.17±16.83)ng/mL、(16.25±1.21)μg/L、(19.52±1.86)μg/L、(22.97±6.45)分,感染组血清FGF23、IL-4、IL-10水平及APACHEⅡ评分均高于未感染组(P<0.05);入院第2天感染组血清Klotho、IFN-γ、IL-2水平分别为(34.95±12.62)ng/mL、(22.19±1.69)μg/L、(28.73±2.95)μg/L,未感染组分别为(51.61±16.08)ng/mL、(25.31±1.74)μg/L、(33.95±1.52)μg/L,感染组血清Klotho、IFN-γ、IL-2水平均低于未感染组(P<0.05);血清FGF23、IL-4、IL-10水平与APACHEⅡ评分呈正相关(相关系数r=0.629、0.597、0.612,P均<0.05),血清Klotho、IFN-γ、IL-2水平与APACHEⅡ评分呈负相关(相关系数r=-0.632、-0.718、-0.701、0.597,P均<0.05);死亡患者入组后1个月血清FGF23、IL-10、IL-4水平分别为(77.49±21.85)ng/mL、(24.76±4.77)μg/L、(24.81±6.28)μg/L,生存患者分别为(58.92±16.94)ng/mL、(18.10±3.82)μg/L、(13.57±4.38)μg/L,死亡患者入组后1个月血清FGF23、IL-10、IL-4水平高于生存患者(P<0.05);死亡患者入组后1个月血清Klotho、IFN-γ、IL-2水平分别为(30.03±11.76)ng/mL、(20.33±2.63)μg/L、(27.19±4.91)μg/L,生存患者分别为(55.68±17.02)ng/mL、(26.54±4.79)μg/L、(35.22±5.64)μg/L,死亡患者入组后1个月血清Klotho、IFN-γ、IL-2水平低于生存患者(P<0.05);血清FGF23、Klotho、IFN-γ、IL-2、IL-4、IL-10预测终末期老年肾病患者发生医院感染、生存预后曲线下面积分别为0.904(OR:0.832~0.953)、0.911(OR:0.840~0.958),且具有良好临床效用(P<0.05)。结论老年终末期肾病患者血清FGF23/Klotho、Th1/Th2细胞因子水平失衡,合并感染患者血清FGF23、IL-10、IL-4水平异常升高,血清Klotho、IFN-γ、IL-2水平表达降低,联合血清FGF23/Klotho、Th1/Th2细胞因子预测医院感染、评估预后的价值较高。

某院儿童肺炎支原体感染情况及23S rRNA基因位点突变与抗生素耐药的相关关系

目的分析某院儿童肺炎支原体感染情况及23S rRNA基因位点突变与抗生素耐药的相关性。方法选取2021年2月至2023年1月在浙江省长兴县中医院就诊的100例疑似肺炎支原体感染患儿,对比不同性别、年龄儿童肺炎支原体的感染情况。根据基因检测结果,将患儿分为突变组和非突变组,并对比2组患儿的临床资料,采用多因素Logistic回归分析影响耐药突变的相关因素。结果100例疑似肺炎支原体感染患儿中,92例(92.0%)PCR检测结果为阳性;其中23S rRNA基因位点突变36例(39.1%),未突变56例(60.9%)。女性患儿的肺炎支原体阳性率(93.2%)稍高于男性(91.1%),且23S rRNA基因位点突变率(41.5%)稍高于男性(37.2%),但差异无统计学意义(P>0.05)。>3岁患儿的肺炎支原体阳性率(96.8%)高于1~3岁患儿(83.8%),且23S rRNA基因位点突变率(47.5%)高于1~3岁患儿(22.6%)(P<0.05)。突变组和未突变组患儿的白细胞计数、嗜中性粒细胞百分比、降钙素原、红细胞沉降率、发热持续时间、咳嗽持续时间、住院时间、本次病程大环内酯类药物应用时间、本次病程肺炎支原体-DNA载量等相比,差异有统计学意义(P<0.05)。多因素Logistic回归分析显示,本次病程肺炎支原体-DNA载量与耐药突变相关(P<0.05)。结论肺炎支原体23S rRNA基因位点突变风险随着肺炎支原体肺炎患儿年龄的增加而逐渐升高。此外,本次病程肺炎支原体-DNA载量与V区A2063G和A2064G耐药突变相关。

癌组织中MACC1、KIF23表达对早期结肠癌术后再发的预测价值

目的观察早期结肠癌患者癌组织内结肠癌转移相关基因-1(MACC1)、驱动蛋白家族成员23(KIF23)表达,并探讨二者对早期结肠癌术后再发的预测价值。方法选择2018年6月至2021年6月拟于新乡市第一人民医院接受腹腔镜联合纤维结肠镜手术的80例早期结肠癌患者,采用实时荧光定量RT-PCR法测定患者癌组织与癌旁组织中MACC1、KIF23 mRNA表达,经Pearson相关性分析MACC1、KIF23 mRNA关系。术后随访2年,根据是否再发分为再发组、未再发组,对比2组术前基础资料与癌组织MACC1、KIF23 mRNA表达,建立Logistic回归模型筛选早期结肠癌术后再发的影响因素,绘制ROC曲线分析术前癌组织MACC1、KIF23 mRNA表达预测早期结肠癌术后再发的价值。结果术前癌组织MACC1 mRNA、KIF23 mRNA相对表达量均高于癌旁组织(P<0.05);Pearson相关性发现,术前癌组织MACC1 mRNA与KIF23 mRNA相对表达量呈正相关(γ=0.326,P<0.05)。再发组肿瘤直径≥3 cm、合并梗阻、TNM分期为Ⅱ期的患者占比及MACC1 mRNA、KIF23 mRNA水平均高于未再发组(P<0.05);经Logistic回归分析,结果显示,肿瘤直径≥3 cm(OR=3.947,95%CI=1.256~12.409)、梗阻(OR=4.385,95%CI=1.273~15.101)、TNM分期Ⅱ期(OR=4.071,95%CI=1.058~15.666)、MACC1 mRNA高表达(OR=2.485,95%CI=1.126~7.325)、KIF23 mRNA高表达(OR=3.467,95%CI=1.298~9.415)是影响早期结肠癌术后再发的独立危险因素(OR>1,P<0.05);绘制ROC曲线发现,术前癌组织MACC1 mRNA、KIF23 mRNA及二者联合预测早期结肠癌术后再发的AUC为0.771(95%CI:0.649~0.894)、0.849(95%CI:0.722~0.976)、0.928(95%CI:0.825~1.000)。结论早期结肠癌患者癌组织内MACC1、KIF23呈高表达,且二者联合可有效预测早期结肠癌术后再发,临床应密切监测患者术前癌组织内MACC1、KIF23的表达水平,以筛查术后再发的高危人群。

亚胺培南耐药鲍曼不动杆菌由插入序列ISAba1点突变引起的bla_(OXA-23)表达减低

目的研究临床分离的8株亚胺培南耐药鲍曼不动杆菌中,与插入序列相关的blaOXA-23 mRNA的表达情况。方法应用双纸片增效法进行金属酶表型筛查并以PCR的方法扩增blaIMP-1、blaIMP-2、blaVIM-2和blaOXA-23 4种β-内酰胺酶基因;实时荧光PCR检测blaOXA-23 mRNA表达量,并以PCR的方法扩增插入序列ISAba1并测序。结果 4株菌金属酶表型筛查为阳性结果,PCR检测8株菌blaOXA-23阳性,2株blaIMP-1阳性、3株blaIMP-2阳性、blaVIM-2基因均为阴性;所有菌株均存在插入序列ISAba1,实时荧光PCR检测显示1株菌blaOXA-23 mRNA表达升高,其余7株菌与对照株比较为表达减低或相近,且这7株菌的插入序列存在点突变现象。结论插入序列ISAba1点突变是引起blaOXA-23碳青霉稀酶活性减低的主要原因。

不同动物种源肺炎克雷伯氏菌分离株23S rRNA序列分析

为分析动物种源肺炎克雷伯氏菌分离株（K．p妇）的进化特点，本研究通过PCR扩增不同动物种源K．pn的23SrRNA基因片段，克隆测序，进行BLAST比对；构建K-pm菌株的系统进化树。结果显示，本实验室保存的12株K．p力核苷酸序列与GenBank收录的X87284株K-pm核苷酸序列同源性为96．8％～99．0％，而与沙门氏菌和奇异变形杆菌同源性只有90．0％～92．7％；8株毛皮动物K．加核苷酸序列同源性为98．1％～99．9％，与猪源、鸡源K．p力的核苷酸序列同源性分别为95．5％～98．1％、96．7％～97．9％；猪源与鸡源K．pn的核苷酸序列同源性为95．9％～97．4％。结果表明，不同动物种源K-pn23SrRNA基因序列同源性差异明显，具有种属的特异性，23SrRNA基因序列可以作为鉴定K-pm的一种快速、简便的方法。

乳腺癌原发灶和淋巴结转移灶中VEGF、nm23的表达变化及意义

目的：观察乳腺癌原发灶及淋巴结转移灶中血管内皮生长因子（VEGF）及转移抑制基因（m23）的表达变化及意义。方法选择2005年1～12月在我院行手术治疗且术前均未行抗癌治疗的60例乳腺癌患者，术后病理证实无同侧腋窝淋巴结转移24例（无转移组），伴有同侧腋窝淋巴结转移36例（转移组），采用免疫组化SP法检测乳腺癌原发灶和淋巴结转移灶中VEGF、nm23的表达，并分析VEGF与nm23的相关性。结果转移组原发灶中VEGF阳性表达率为91．7％，转移灶中为61．1％，无转移组VEGF阳性表达率为70．8％，转移组原发灶中VEGF阳性表达率明显高于转移灶及无转移组（P均＜0．05）。转移组nm23阳性表达率在原发灶中为16．7％，转移灶中为41．7％，无转移组nm23阳性表达率为45．8％，转移组原发灶中nm23阳性表达率明显低于转移灶及无转移组（P均＜0．05）。原发灶VEGF与nm23表达呈负相关（r＝-0．415，P＜0．05）。原发灶VEGF、nm23表达与淋巴结转移灶中VEGF、nm23表达无相关性。结论乳腺癌原发灶中VEGF表达明显高于、nm23表达明显低于淋巴结转移灶， VEGF、nm23与乳腺癌淋巴结转移密切相关，二者在乳腺癌发生和淋巴结转移中起拮抗作用。