Value of subtypes of antimitochondrial antibodies in primary biliary cholangitis / 中华风湿病学杂志

Objective:To investigate the distribution and clinical significance of subtypes of antimitochondrial antibodies (AMA)-M2, M4, M9 in primary biliary cholangitis (PBC).Methods:A total of 1 367 patients were detected with AMA-M2, M4, M9 in Peking Union Medical College Hospital (PUMCH) from Jan 2014 to Dec 2019 and the clinical parameters were collected. The distribution patterns of AMA subtypes in different groups were analyzed and the diagnostic sensitivity and specificity of AMA subtypes in PBC were calculated. Chi-square test was used for statistical analysis.Results:In 1 367 patients, 236 of whom were positive for AMA subtypes. The positivity of AMA subtypes in female was significantly higher than in male (20.34% vs 9.41%, χ2=23.792, P<0.01). In addition, the positivity of AMA subtypes was significantly higher in 30-65 years old patients than in patients younger than 30 years old or older than 65 years old [(20.00%(193/965) vs 10.97%(17/155) vs 10.53%(26/247), χ2=17.209, P<0.01]. 110 patients with positive AMA subtypes were diagnosed with PBC. The diagnostic sensitivity and specificity of AMA-M2 were both desirable [94.64%(106/112) and 92.35%(1 159/1 255)]. Although the specificity of AMA-M4 was as high as 99.12%(1 244/1 255), its sensitivity was very low [15.18%(17/122)]. Combined detection of different AMA subtypes could not improve the diagnostic sensitivity and specificity significantly. Diseases other than PBC can be positive for AMA subtypes, predominantly for AMA-M2. Conclusion:Female and 30-65 years old patients were more frequently positive for AMA subtypes. AMA-M2 was the most valuable AMA subtype for diagnosing PBC.

Ratio analysis of ERβ isoforms in paired cancerous and adjacent normal breast tissues / 肿瘤研究与临床

Objective To illustrate the composition ratio of ERβ isoforms in paired cancerous and adjacent normal tissues from breast cancer patients.Methods Eighty-seven pairs of cancerous and adjacent normal tissues were obtained from breast cancer patients.RT-qPCR was used to determine the relative mRNA expression levels of ERβ isoforms (ER[β1,ERβ2 and ERβ5),and the composition ratios of ERβ isoforms were analyzed.Results The expression levels of all tested ERβ isoforms (ERβ1,ERβ2 and ERβ5) in breast cancer tissues were much lower than those in adjacent normal breast tissues (P ＜ 0.01).Isoform ratio analysis showed that ERβ5 was the dominant isoform in both cancerous and adjacent normal tissues with a positive detection rate of 54.02 ％ and 75.84 ％,respectively.Meanwhile,ERβ1 had the lowest detection rate (9.74 ％ and 6.77 ％ in cancerous and adjacent normal tissues,respectively).The positive rates for both ERβ1 and ERβ2 were much lower in adjacent normal tissues than those in cancer tissues (Z =-2.24,P =0.025 and Z =-4.85,P ＜ 0.01,separately),while more cancerous tissues were ERβ5-positive in comparison to adjacent normal tissues (Z =-5.32,P ＜ 0.01).Conclusions The expression levels of all the ERβ isoforms are differentially down-regulated with significant alterations in their composition ratios during breast carcinogenesis.Further understanding on molecular mechanisms underlying the differential down-regulation of ER[β isoforms will shed new light on breast carcinogenesis.

The relationship of breast cancer molecular subtype and location and time of recurrence metastasis / 中国医师杂志

Objective To investigate the risk distribution of breast cancer for location and time of recurrence metastasis in molecular subtype.Methods We studied retrospectively the female patients who were diagnosed as invasive ductal breast cancer in our hospital from July 2004 to June 2012,detected ER,PR,and HER2 expressions in the paraffin sections.The patients with recurrence metastasis were divided into local recurrence and distant metastasis with the first transfer site as standard for analyzing the distribution in molecular subtype and the time of the first site of recurrence metastasis.Results Sixty two patients were encountered recurrence metastasis,including 23 patients with local recurrence,and 39 patients with distant metastasis,death 11.The rates of distant metastasis for patients who belonged to HER2 type and basal-like type were higher than that of local recurrence (P =0.01,P =0.001).The risk distribution of recurrence metastasis time in molecular recurrence metastasis showed that 35 percent of recurrence metastasis time of luminal A type was first 3 years,75 percent of molecular subtype of basa1-1ike type recurrence metastasis time in first 3 years and advanced.The peak of luminal B and HER2 type was first 3 years,and very low in 5 years.Conclusions Molecular subtype of breast cancer is an important complement for TNM method in accurately assessing the patients of recurrence metastasis for location and time,and is helpful for the individual screening of patients for recurrence metastasis.

Papel da titina na modulação da função cardíaca e suas implicações fisiopatológicas / The role of titin in the modulation of cardiac function and its pathophysiological implications / Papel de la Titina en la Modulación de la Función Cardíaca y sus Implicaciones Fisiopatológicas

A titina é uma proteína sarcomérica gigante que se estende desde a linha Z até a linha M. Em razão de sua localização, representa um importante sensor biomecânico com um papel fundamental na manutenção da integridade estrutural do sarcômero. A titina funciona como uma "mola bidirecional" que regula o comprimento sarcomérico e realiza ajustes adequados da tensão passiva sempre que o comprimento varia. Dessa forma, não só determina a rigidez ventricular e a função diastólica, como também influencia a função cardíaca sistólica, modulando o mecanismo de Frank-Starling. O miocárdio expressa duas isoformas dessa macromolécula: a N2B, mais rígida, e a isoforma N2BA, mais complacente. As alterações na expressão relativa das duas isoformas da titina ou alterações do seu estado de fosforilação têm sido implicadas na fisiopatologia de várias doenças como a insuficiência cardíaca diastólica, a cardiomiopatia dilatada, a cardiomiopatia isquêmica e a estenose aórtica. Neste artigo pretende-se descrever sumariamente a estrutura e localização da titina, a sua relação com diferentes cardiomiopatias, e compreender de que forma as alterações dessa macromolécula influenciam a fisiopatologia da insuficiência cardíaca diastólica, salientando o potencial terapêutico da manipulação dessa macromolécula.

Titin is a giant sarcomeric protein that extends from the Z-line to the M-line. Due to its location, it represents an important biomechanical sensor, which has a crucial role in the maintenance of the sarcomere structural integrity. Titin works as a "bidireactional spring" that regulates the sarcomeric length and performs adequate adjustments of passive tension whenever the length varies. Therefore, it determines not only ventricular rigidity and diastolic function, but also systolic cardiac function, modulating the Frank-Starling mechanism. The myocardium expresses two isoforms of this macromolecule: the N2B, more rigid and the isoform N2BA, more compliant. The alterations in the relative expression of the two titin isoforms or alterations in their state of phosphorylation have been implicated in the pathophysiology of several diseases, such as diastolic heart failure, dilated cardiomyopathy, ischemic cardiomyopathy and aortic stenosis. The aim of this study is to describe, in brief, the structure and location of titin, its association with different cardiomyopathies and understand how alterations in this macromolecule influence the pathophysiology of diastolic heart failure, emphasizing the therapeutic potential of the manipulation of this macromolecule.

La titina es una proteína sarcomérica gigante que se extiende desde la línea Z hasta la línea M. En razón de su ubicación, representa un importante sensor biomecánico con un papel fundamental en la manutención de la integridad estructural del sarcómero. La titina funciona como un "resorte bidireccional" que regula el largo sarcomérico y realiza ajustes adecuados de la tensión pasiva siempre que ese largo varía. De esa forma, no sólo determina la rigidez ventricular y la función diastólica, sino también influye en la función cardíaca sistólica, modulando el mecanismo de Frank-Starling. El miocardio expresa dos isoformas de esa macromolécula: la N2B, más rígida, y la isoforma N2BA, más complaciente. Las alteraciones en la expresión relativa de las dos isoformas de la titina o alteraciones de su estado de fosforilación han sido implicadas en la fisiopatología de varias enfermedades como la insuficiencia cardíaca diastólica, la cardiomiopatía dilatada, la cardiomiopatía isquémica y la estenosis aórtica. Este artículo pretende describir sumariamente la estructura y ubicación de la titina, su relación con diferentes cardiomiopatías, y comprender de qué forma las alteraciones de esa macromolécula influyen en la fisiopatología de la insuficiencia cardíaca diastólica, destacando el potencial terapéutico de la manipulación de esa macromolécula.

Detection of genotypes of plasmid-mediated quinolone resistance in clinical isolates of Shigella / 中华传染病杂志

ObjectiveTo investigate the variations and distributions of the plasmid-mediated quinolone resistance genes in clinical isolates of Shigella and their resistance to antimicrobial agents. Methodsqnr, aac(6')-Ib-cr and qepA genes were identified by polymerase chain reaction (PCR) in 137 clinical isolates of Shigella.DNA sequencing of gene-positive strains were analyzed and the conjugation experiment was performed. The minimal inhibitory concentrations (MIC) of Shigella isolates, recipient strains and transconjugants were tested by agar dilution method for quinolones and other antimicrobial agents. The genotype of transconjugants were determined by PCR and sequencing. ResultsFour (2.9％) strains of the 137 Shigella isolates were qnr gene positive, including 3 qnrS2 positive and 1 qnrB4 positive (GenBank accession numbers of the complete sequence were JF261185 and HQ917003, respectively).Furthermore,five (3.6％) aac ( 6')-Ib-cr gene-positive strains (GenBank accession number JF261186 ) and one (0.7％) qepA gene-positive strain were identified in all isolates. The conjugation experiments were successfully carried out in six out of ten PCR-positive isolates. The MIC of transconjugants against quinolones and other antimicrobial agents increased differently compared to recipient strains. Conclusions The plasmid-mediated quinolone resistance genes are lowly prevalent in clinical isolates of Shigella. However, these resistance genes have the characteristic of horizontal transfer, which indicates that more attention should be paid to this phenomenon.

Quantitative detection of prostate specific membrane antigen and the splicing variant DNA in different tumor cell strains and prostate tissues / 中华老年医学杂志

Objective To understand and illuminate the bionomic characteristics of prostate specific membrane antigen (PSMA) and splicing variant PSMA5, through detecting the DNA levels of them in different tumor cell strains and prostate tissues. Methods The fluorescent quantization reverse transcriptase PCR (FQ-RT-PCR) method built up by our research group was used to detect the PSMA and variant PSMA5 DNA levels in different tumor cell strains and prostate tissues. Results The PSMA and PSMA5 DNA levels in tumor cell strains and pathological prostatic tissues were obviously more than those of the normal prostatic tissues (F=3.40, 11.94, both P＜0.05), and the PSMA5 DNA level was much higher than was the PSMA DNA level in prostatic carcinoma tissues (P＜0.05). Conclusions The different expressions between PSMA and PSMA5 in different tumor cells and prostatic tissues show that PSMA5 is more specific than PSMA as a prostate carcinoma tumor marker.

Genetic barriers of primary drug resistance mutations in 123 recombinant subtype strains of human immunodeficiency virus-1 / 中华传染病杂志

Objective To compare the genetic barriers to development of primary mutations related to drug resistance to protease inhibitors (PI), nucleioside reverse transcriptase inhibitors ( NRTI ), and non-nucleioside reverse transcriptase inhibitors ( NNRTI ) among human immunodeficiency virus (HIV)-1 CRF01_AE, CRF07_BC, and CRF08_BC strains, and to understand the difference of varying patterns of drug resistance related mutations within these subtypes. Methods One hundred and ninety naive HIV-positive subjects from Nanning City and Liuzhou City, Guangxi Zhuang Autonomous Region, were recruited. Peripheral blood samples were collected from all participants. HIV-1 RNAs were extracted from plasma, and the pol regions were amplified and sequenced. Sequences were subjected to phylogenetic analysis to determine the subtypes of HIV-1 isolates. Nucleotide transitions and transversions were counted for each primary mutation in these sequences. According to the phenomena that transitions occur on average 2. 5 times frequently than transversions, each transition was scored as 1, and each transversion scored as 2. 5. The sum of the scores for a particular substitution was calculated, and this value was taken as the genetic barrier to development of this mutation. Then, the differences of genetic barriers among the subtypes were assessed by Kruskal-Wallis test and Nemenyi test. Results A total of 123 sequences of CRF01_AE,CRF07_BC and CRF08_BC strains were selected. CRF08_BC had a lower genetic barrier for T/S69Dsubstitution than CRF01_AE and CRF07_BC (χ2 =107. 501, P＜0.01), while CRF01_AE and CRF07_BC had lower genetic barriers for V118I and L210W substitution than CRF08_BC. In addition,CRF07_BC had a decreased genetic barrier for V106M compared with CRF01_AE and CRF08_BC.Conclusions In the presence of the same selective pressure, subtypes CRF01_AE and CRF07_BC may be more likely to develop V118I and L210W substitution than CRF08_BC. However, CRF08_BC may be more likely to develop T/S69D substitution than CRF01_AE and CRF07_BC. Meanwhile, CRF07_BC may be easier to develop V106M substitution than CRF01_AE and CRF08_BC.

Analysis of haemagglutinin gene of the first influenza A-H1N1 viral strain isolated in Guangdong Province during an influenza A pandemic in 2009 / 中华传染病杂志

Objective To detect and analyze the haemagglutinin (HA) gene of the first influenza A-H1N1 viral strain isolated in Guangdong Province during an influenza A pandemic in 2009.Methods A-H1N1 virus strain was isolated from the throat swab of the first patient diagnosed with A-H1N1 virus infection in Guangdong Province in 2009. Viral nucleonic acid was extracted from supernatant of cell culture and amplified using reverse transcriptase-polymerase chain reaction (RT-PCR) with HA gene-specific primers. The product was cloned, sequenced, and the homology was analyzed. Results A 1710 bp HA gene of the first influenza A-H1N1 viral strain in Guangdong Province in 2009 was acquired, which was named as A/GuangzhouSB/01/2009 (H1N1) HA with GenBank access No. GQ268003. The homology of the studied HA gene and the 277 influenza A (H1N1) isolates reported in the epidemic areas was 99.0%-99.8%, and as high as 99.8% when compared with the isolates reported in the United States where the patient had traveled. When the studied HA gene was compared with 25 isolates of Chinese seasonal A-H1N1 virus, the homology was 72.3%-85.6%. Conclusions The homology of the first isolated A-H1N1 viral strain in Guangdong Province in 2009 and epidemic influenza A-H1N1 virus is high, while it is low compared with Chinese seasonal A-H1N1 virus.

Adaptation of cAMP signaling system in SH-SY5Y neuroblastoma cells following expression of a constitutively active stimulatory G protein alpha, Q227L Gsalpha

Heterotrimeric GTP-binding proteins (G protein) are known to participate in the transduction of signals from ligand activated receptors to effector molecules to elicit cellular responses. Sustained activation of cAMP-G protein signaling system by agonist results in desensitization of the pathway at receptor levels, however it is not clear whether such receptor responses induce other changes in post-receptor signaling path that are associated with maintenance of AMP levels, i.e. cAMP-forming adenylate cyclase (AC), cAMP-degrading cyclic nucleotide phosphodiesterase (PDE) and cAMP-dependent protein kinase (PKA). Experiments were performed to determine the expression of AC, PDE, and PKA isoforms in SH-SY5Y neuroblastoma cells, in which cAMP system was activated by expressing a constitutively activated mutant of stimulatory G protein (Q227L Gsalpha). Expression of ACI mRNA was increased, but levels of ACVIII and ACIX mRNA were decreased. All of the 4 expressed isoforms of PDE (PDE1C, PDE2, PDE 4A, and PDE4B) were increased in mRNA expression; the levels of PKA RIalpha, RIbeta, and RIIbeta were increased moderately, however, those of RIIalpha and Calpha were increased remarkably. The activities of AC, PDE and PKA were also increased in the SH-SY5Y cells expressing Q227L Gsalpha. The similar changes in expression and activity of AC, PDE and PKA were observed in the SH-SY5Y cells treated with dbcAMP for 6 days. Consequently, it is concluded that the cAMP system adapts at the post-receptor level to a sustained activation of the system by differential expression of the isoforms of AC, PDE, and PKA in SH-SY5Y neuroblastoma. We also showed that an increase in cellular cAMP concentration might mediate the observed changes in the cAMP system.