OBJECTIVE: Porcine stress syndrome (PSS) is one kind of molecular genetics defect diseases of pig which will cause malignant hyperthermia syndrome (MHS) and is the first index should be excluded in screening of a pig species for xenotransplantation. It was reported that mutation of pig rynodine receptor(RYR1) gene is the main reason for PSS. In this study, RYR1 genotypes of the Chinese Banna mini pig inbred line and inbreeding closed colony Wuzhishan pig were investigated with polymerase chain reaction-restriction endonuclease fragment length polymorphism (PCR-RFLP) technique. METHODS: Antevenocaval whole blood samples were collected from 50 Banna mini-pig inbred-line(BMI), 15 inbreeding Wuzhishan pig (WZSP) and 25 Neijiang pigs (NJP) as negative control, the primer were designed and synthesized, PCR reaction was conducted following the sequence of 94 degrees C (1 min), 58 degrees C (1 min) and 72 degrees C (1 min) for 30 cycles. The PCR products were digested with restriction endonuclease HhaI and then electrophoresis check. RESULTS: A 659 bp DNA fragment was amplified with these two primers, the HALNN sample fragment was cut into fragments as 493 bp and 166 bp individually after the digestion, indicates no point mutation at site 1,843 in RYR1 gene in all tested BMI pig and WZSP. Namely, the RYR1 genotype of 50 cases of BMI and 15 cases of WZSP were HALNN, therefore their phenotype is PSS negative. CONCLUSION: It indicates that the genotype of Banna mini pig inbred line and inbreeding Wuzhishan pig are HALNN therefore PSS absolutely negative, the group penetrance is 0. This is consistent with experimental observation. It suggests that Banna mini pig inbred line and inbreeding Wuzhishan pig may be the alternative donor for xenotransplantation.
The present study was aimed to explore the relationship of transforming growth factor (TGF) β3 gene SfaNI polymorphism (rs3917201 locus) and non-syndromic cleft lip with or without cleft palate (NSCL/P) in people of the Uygur's Nationality and Han's in Xinjiang, China. TGFβ3 gene fragment including SfaNI was amplified and purified as the template of the primer extension reaction thenafter. The single base extension reaction was carried out using SNP specific extension primer. The products were purified and analyzed by MALDI-TOF. The test showed that there were not significantly different frequencies of AA, AG, GG genotypes and alleles between the whole NSCL/P group and the whole control group (P>0.05).Within the Uygurs or Hans, the frequencies of genotypes between the whole NSCL/P group and the whole control group were not significantly different(P>0.05). The distributions of the A, G alleles between the NSCL/P group and the control group were not significantly different within the Uygurs (P>0.05), but significant different within the Hans (P<0.05). In all the NSCL/P patients, frequencies of genotypes and alleles were not significantly different between Uygurs and Hans (P>0.05), and not significantly (P>0.05) either between Uygurs and Hans in all the healthy persons. The results proved that TGFβ3 gene SfaNI polymorphism may not be related to NSCL/P in Xinjiang Uygur people, while the occurrence of NSCL/P in Han population may be related to frequency of the A and G allele of SfaNI polymorphism.
Online Mendelian Inheritance in Man (OMIM) is a knowledge source and data base for human genetic diseases and related genes. Each OMIM entry includes clinical synopsis, linkage analysis for candidate genes, chromosomal localization and animal models, which has become an authoritative source of information for the study of the relationship between genes and diseases. As overlap of disease symptoms may reflect interactions at the molecular level, comparison of phenotypic similarity may indicate candidate genes and help to discover functional connections between genes and proteins. However, the OMIM has used free text to describe disease phenotypes, which does not suit computer analysis. Standardization of OMIM data therefore has important implications for large-scale comparison of disease phenotypes and prediction of phenotype-genotype correlations. Recently, standard medical language systems, term frequency-inverse document frequency and the law of cosines for document classification have been introduced for mining of OMIM data. Combined with Gene Ontology and various comparison methods, this has achieved substantial successes. In this article, we have reviewed various methods for standardization and similarity comparison of OMIM data. We also predicted the trend for research in this direction.
ObjectiveTo clarify the relationship between the G/C polymorphism of inflammatory gene matrix metalloproteinase-2 (MMP2) and warfarin therapy after cardiac valve replacement (CVR). MethodsWe finally identified 96 patients who received additional warfarin therapy after CVR as a trial group and 78 patients without the warfarin therapy as a control group. Gene sequencing techniques were adopted to determine single nucleotide polymorphism allele. We analyzed genotype and clinical features of the two groups and explored the relationship between the different MMP2 geno-types and warfarin therapy after CVR. Logistic regression was used to analyze the correlation between genotypes and risk factors after CVR and Kaplan-Meier survival curves were performed to analyze the survival time and efficacy of patients carrying MMP2 GC and GG genotypes. ResultsThe distribution of MMP2 genotype in patients receiving warfarin therapy after surgery was different from that in patients without warfarin therapy. The results of multivariate logistic regression analysis showed that GC and GG genotypes were risk factors of complications of CVR. The proportion of GG genotype was higher in the patients with postoperative complications compared with those without. The survival time of patients carrying genotype MMP2 GG was shorter than those carrying GC genotype (P < 0.05), which reveals that the level of MMP2 GG genotype was associated with the prognosis. ConclusionG allele of MMP2 is a risk factor of complications following CVR. GG genotype is relevant to CVR and prognosis, which can be regarded as a risk factor post CVR.
ObjectiveTo analyze the drug-resistant phenotype and genotype characteristics of carbapenem-resistant Enterobacteriaceae (CRE) in a traditional Chinese medicine hospital from 2016 to 2018, to provide guidance for clinical rational drug use and effective anti-infection treatment.MethodsA total of 2 901 Enterobacteriaceae bacteria strains isolated from January 2016 to December 2018 were selected, and CRE strains were screened by microdilution test and Kirby-Bauer methods. CRE strains with successful seed preservation and detailed clinical data were selected for carbapenemase phenotype confirmation test, drug-resistant gene amplification, and sequencing comparison.ResultsThe 101 CRE strains collected between 2016 and 2018 were mainly Klebsiella pneumonia (73.27%, 74/101) and Escherichia coli (14.85%, 15/101), and the specimens were mainly from sputum (63.37%, 64/101) and catheter urine (11.88%, 12/101). The phenotypic test results of carbapenemase showed that 94 strains were positive in modified Hodge test, with a positive rate of 93.07%, 96 strains were positive in Carba NP test, with a positive rate of 95.05%, and 98 strains were positive in modified carbapenem inactivation method test, with a positive rate of 97.03%. Drug-resistant genes were detected in 92 (91.01%) of the 101 CRE strains, sequencing results showed that 66 (65.35%) carried blaKPC-2 gene, 4 (3.96%) carried blaKPC-19 gene, 9 (8.91%) carried blaNDM-1 gene, and 13 (12.87%) carried blaNDM-5 gene. No CRE strains carrying two resistance genes were detected. Among them, Klebsiella pneumoniae strains mainly carried blaKPC-2 gene (82.43%, 61/74), and Escherichia coli strains mainly carried blaNDM-5 gene (86.67%, 13/15), which were consistent with the main epidemic genotype in China.ConclusionsIn recent three years, the CRE strains in this hospital mainly included Klebsiella pneumoniae with blaKPC-2 gene and Escherichia coli with blaNDM-5 gene. According to the results of this test, we can reasonably select antimicrobial agents in combination with the drug sensitivity report from the microbial laboratory, so as to delay the growth of drug-resistant strains and prevent hospital transmission of multidrug-resistant bacteria.
ObjectiveTo summarize the research status of the relationship between hepatitis B virus X protein (HBx), hepatitis B virus (HBV) genotypes and hepatocellular carcinoma (HCC) at home and abroad, and to prospect its clinical significance.MethodThe literatures about HBx, HBV genotypes and HCC were reviewed.ResultsThere was a close relationship between HBx and the occurrence, development, migration and metastasis of HCC. There was a certain association between HBV genotypes and HCC, but the specific mechanism had not been clarified.ConclusionsHBx and HBV genotypes play an important role in the occurrence and development of HCC. With the further study of molecular mechanism, it will promote the diagnosis and treatment of hepatitis B, liver cirrhosis and liver cancer, and provide more individualized intervention for clinical workers.
Objective To explore the clinical characteristics of patients who were infected by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) of different genotypes. Methods A retrospective study was conducted on 111 SARS-CoV-2 infected cases at home and abroad admitted to Chengdu Public Health Clinical Medical Center between January and September 2020. The basic information, gene sequencing results (Pangolin typing method), clinical typing, first laboratory examinations 24 hours after admission, and whether repositive after discharge were collected. According to Pangolin typing, patients were divided into five groups: A, B, B.X, B.1.X and B.1.1.X. The basic information (age, sex, and origin), laboratory test results (lymphocyte count, C-reactive protein, serum amyloid A, CD3+ T lymphocytes, CD4+ T lymphocytes, and CD8+ T lymphocytes), clinical classification and whether repositive were compared among different genotype infected patients. Results Among the 111 infected patients, 54 (48.6%) were males and 57 (51.4%) were females. Their ages ranged from 16 to 87 years, with a median age of 49 years. In terms of clinical classification, there were 10 asymptomatic cases (9.0%), 10 mild cases (9.0%), 64 ordinary cases (57.7%), 13 severe cases (11.7%), and 14 critical cases (12.6%). There were 75 domestic cases (67.6%) and 36 imported cases (32.4%). Eighty cases (72.1%) did not return to positive, and 31 cases (27.9%) returned to positive. There were 8 cases infected by type A virus, 18 cases infected by type B virus, 26 cases infected by type B.X virus, 5 cases infected by type B.1.X virus, and 54 cases infected by type B.1.1.X virus. Among patients infected by different genotype viruses, no statistically significant difference was found in sex, age, clinical type, laboratory examination, or whether repositive (P>0.05), but there was statistically significant difference in the distribution of domestic and imported cases (P=0.016). Type B virus infected patients were mostly domestic cases, while type B.X virus infected patients were mostly imported cases. Conclusion The distribution of domestic and imported cases is different among SARS-CoV-2 of different genotypes.
Objective To explore the colonization of Klebsiella pneumoniae in the intensive care unit of our hospital and analyze the risk factors. Methods A total of 226 patients were actively screened in the surgical intensive care unit and neurosurgery intensive care unit from June to December 2020 in the hospital, and their clinical data were retrospectively analyzed. Results Totally, 87 strains of Klebsiella pneumoniae were screened out, 69 strains were carbapenem-resistant Klebsiella pneumoniae (CRKP), and the resistant genotype was mainly KPC genotype (79.6%). The resistance rates of meropenem were 75.0% and 77.4%, respectively. Age and pulmonary infection before admission are risk factors for CRKP colonization, while pulmonary infection before admission is an independent risk factor for CRKP colonization. Conclusions Both the CRKP colonization rate of patients and the rate of resistance to carbapenem antimicrobials are relatively high in the intensive care unit of our hospital. Pulmonary infection before admission is an independent risk factor for CRKP colonization.