Comportamiento alimentario de Aedes aegypti en brotes de dengue de dos zonas rurales del Perú durante el ciclón Yaku y El Niño Global del 2023 / Aedes aegypti feeding behavior during dengue outbreaks in two rural areas of Peru during the Yaku cyclone and El Niño phenomenon of 2023

RESUMEN Objetivos. Determinar la alimentación del Aedes aegypti en brotes de dengue de dos zonas rurales del Perú durante el ciclón Yaku y El Niño Global del 2023. Material y métodos. Se analizaron ocho muestras de sangre (8 pooles) obtenidas del abdomen de 80 especímenes Aedes aegypti capturados en los distritos rurales de Querecotillo y Marcavelica durante brotes de dengue acontecidos en el ciclón Yaku y en El Niño Global. Se extrajo ADN de las muestras analizadas, se llevó a cabo una PCR dirigida al gen CytB como marcador genético y los productos PCR fueron digeridos enzimáticamente con las restrictasas Hae III y Mwo I. Los productos PCR-RFLP fueron visualizados por electroforesis en gel de agarosa al 4%. Resultados. Se obtuvo ADN de todas las muestras y como producto PCR un amplicón de 358 pb. Así mismo, el único RFLP en Hae III observado fue el de Homo sapiens sapiens (233 y 125 pb). No se observó RFLP en Hae III de Gallus gallus y RFLP en Mwo I de Canis familiaris y Mus musculus. Conclusión. En brotes de dengue de zonas rurales, durante el ciclón Yaku y en El Niño Global, el Aedes aegypti presentó un comportamiento alimenticio antropofílico conservado.

ABSTRACT Objective. To determine the feeding behavior of Aedes aegypti in dengue outbreaks in two rural areas of Peru during the Yaku cyclone and El Niño phenomenon of 2023. Material and methods. Eight blood samples (8 pools) were obtained from the abdomen of 80 Aedes aegypti specimens captured in the rural districts of Querecotillo and Marcavelica during the Yaku cyclone and El Niño dengue outbreaks. DNA was extracted from the analyzed samples, then a PCR was directed at the CytB gene as a genetic marker and the PCR products were enzymatically digested with the restrictases Hae III and Mwo I. The PCR-RFLP products were visualized by agarose gel electrophoresis at 4%. Results. DNA was obtained from all samples and a 358 bp amplicon was obtained as a PCR product. Likewise, the only RFLP found in Hae III was from Homo sapiens sapiens (233 and 125 bp). RFLP was not found in Hae III of Gallus gallus and RFLP in Mwo I of Canis familiaris and Mus musculus. Conclusion. Aedes aegypti showed conserved anthropophilic feeding behavior in dengue outbreaks in rural areas during the Yaku cyclone and El Niño.

Blood lipids and apolipoprotein E gene in patients with cardiovascular and cerebrovascular diseases in Honghe Hani and Yi Autonomous Prefecture of Yunnan Province / 中国基层医药

Objective:To analyze the distribution of blood lipids and apolipoprotein E ( ApoE) gene in patients with cardiovascular and cerebrovascular diseases in Honghe Hani and Yi Autonomous Prefecture of Yunnan Province, and their relationship with ethnicity, gender, and age. Methods:A total of 102 patients with cardiovascular and cerebrovascular diseases who received treatment in Southern Central Hospital of Yunnan Province (The First People's Hospital of Honghe State) from May 2019 to June 2020 were included in this study. Their blood lipid and ApoE gene distributions were analyzed. The correlations between blood lipid and ApoE gene distribution and ethnicity, gender, and age were analyzed. Results:There was a significant difference in total cholesterol level between male and female patients with cardiovascular and cerebrovascular diseases in Honghe Hani and Yi Autonomous Prefecture of Yunnan Province [(4.10 ± 1.27) mmol/L vs. (4.70 ± 1.83) mmol/L, t = 1.87, P = 0.048]. There was no significant difference in total cholesterol level among different ethnic groups and different ages ( P = 0.343, 1.000). There were no significant differences in low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, and triglyceride levels among different ethnic groups, different sexes, and different ages ( P = 0.562, 0.125, 0.158; 0.884, 0.068, 0.681; 0.262, 0.367, 0.965). There were no significant differences in blood lipid levels among different ethnic groups, different sexes, and different ages ( P = 0.890, 0.336, 0.142). No E2/E4 gene was found in all patients. E2/E2 and E2/E3 genes of ApoE accounted for 16.67%, E3/E3 gene accounted for 70.59%, and E3/E4 and E4/E4 genes accounted for 12.74%. There was no significant difference in the distribution of ApoE gene among different ethnic groups, different sexes, and different ages ( χ2 = 0.13, 0.69, 0.44, P = 0.936, 0.429, 0.804). Conclusion:Female patients with cardiovascular and cerebrovascular diseases in Honghe Hani and Yi Autonomous Prefecture of Yunnan Province have higher total cholesterol levels than male patients. Low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, and triglyceride levels are not significantly correlated with ethnicity, sex, and age. Blood lipid abnormalities mainly manifest as low high-density lipoprotein cholesterol levels and high triglyceride levels. The E2/E4 gene has not been found in this region so far.

Analysis and clinical significance of cytochrome P 450 2C19 gene in patients with cardiovascular and cerebrovascular diseases in southern Yunnan / 中国基层医药

Objective:To analyze the distribution and clinical significance of cytochrome P 450 2C19 (CYP2C19) gene in patients with cardiovascular and cerebrovascular diseases in southern Yunnan. Methods:The data of 245 patients with cardiovascular and cerebrovascular diseases who received treatment in Southern Central Hospital of Yunnan Province between May 2019 and June 2020 were retrospectively analyzed. The distribution of CYP2C19 gene and its relationship with nationality, age, sex, blood lipids, hypertension, and diabetes were analyzed and compared between southern Yunnan and other regions.Results:The proportions of seven phenotypes of CYP2C19 gene *1/*17, *1/*1, *1/*2, *1/*3, *2/*2, *2/*3, *3/*3 in 245 patients were 2.86%, 38.37%, 39.18%, 5.31%, 9.39%, 4.08% and 0.82%, respectively. The proportions of individuals with superfast/ultrafast metabolism, fast metabolism, intermediate metabolism, and slow metabolism in 245 patients were 2.86%, 38.37%, 44.49%, and 14.29%, respectively. The frequency of polymorphisms in the CYP2C19 gene was consistent with the Hardy-Weinberg equilibrium ( P > 0.05), which was constant and representative. The Fisher test showed that the CYP2C19 gene distribution of patients with cardiovascular and cerebrovascular diseases in southern Yunnan was not greatly correlated with nationality, age, sex, underlying disease, blood lipids, and the types of cardiovascular and cerebrovascular diseases (all P > 0.05). There was a significant difference in CYP2C19 gene distribution in patients from southern Yunnan versus Dongguan, Jiangxi, Fujian, northern Sichuan, Chifeng, Xiamen, Shaanxi, and Kunming ( P < 0.001, < 0.001, 0.045, 0.008, 0.001, 0.005, < 0.001, 0.016). Conclusion:The distribution of CYP2C19 gene in patients with cardiovascular and cerebrovascular diseases in southern Yunnan is not obviously correlated with nationality, age, sex, underlying diseases, blood lipids, and the types of cardiovascular and cerebrovascular diseases. CYP2C19 gene distribution is related to regional distribution, which can guide personalized medication in different regions.

Analysis of solute carrier organic anion transporter family member 1b1 and apolipoprotein E genes in a population from southern Yunnan / 中国基层医药

Objective:To analyze the distribution of solute carrier organic anion transporter family member 1b1 ( SLCO1B1) and apolipoprotein E ( ApoE) genes in a population from southern Yunnan. Methods:The data of 104 patients who received treatment in Southern Central Hospital of Yunnan Province (The First People's Hospital of Honghe State) between May 2019 and June 2020 were collected. The distribution of SLCO1B1 and ApoE genes and their relationship with nationality, sex, and age were analyzed and compared between different regions. Results:The percentage of patients carrying *1a/*1a, *1a/*1b, *1b/*1b, *1a/*15, *1b/*15, five phenotypes of SLCO1B1 gene, in the population from southern Yunnan was 4.81%, 32.69%, 42.31%, 12.50% and 7.69% respectively. Phenotypes *1a/*5, *5/*5, *5/*15 and *15/*15 were not detected. Normal metabolic phenotype of SLCO1B1 accounted for 79.81%, and intermediate metabolic phenotype of SLCO1B1 accounted for 20.19%. Weak metabolic phenotype was not detected. The percentage of patients carrying E2/E2, E2/E3, E3/E3, E3/E4, E4/E4, five phenotypes of ApoE gene in the population from southern Yunnan was 0.96%, 16.35%, 70.19%, 11.54% and 0.96% respectively. E2/E4 phenotype was not detected. The percentage of patients with ApoE protective phenotype, ApoE normal phenotype, and ApoE risk phenotype was 17.31%, 70.19% and 12.50% respectively. The observed polymorphism mutation frequency of SLCO1B1 and ApoE genes was consistent with the Hardy-Weinberg equilibrium ( P > 0.05), suggesting constancy and a population representation. The Fisher test showed that SLCO1B1 gene distribution differed significantly between ethnic minorities and Han nationality in southern Yunnan ( P = 0.013). There was no significant difference in SLCO1B1 gene distribution between different sexes and between different ages (all P > 0.05). There was no significant difference in ApoE gene distribution between ethnic minorities and Han nationality, between different sexes, and between different ages in the population from southern Yunnan (all P > 0.05). Conclusion:SLCO1B1 gene distribution is related to nationality in the population from southern Yunnan, but it is unrelated to sex and age. ApoE gene distribution is unrelated to nationality, sex and age.

Metallothionein-2a (rs1610216&rs28366003) gene polymorphisms and the risk of stomach adenocarcinoma / Polimorfismos do gene metalotioneína-2A (rs1610216& rs28366003) e o risco de adenocarcinoma de estômago

ABSTRACT BACKGROUND: Gastric cancer is the fourth most common cause of worldwide cancer. Also in contrast to the huge advances in curing, the chance of living is very low even in surgery cases. Having a genetic predisposition plays an important role in cancer development. The association between Metallothionein-2A gene polymorphisms and the risk of adenocarcinoma has been widely studied, yet there is only one study on stomach diseases. OBJECTIVE: In this study, we aimed to investigate the association between 2 (MT-2A) polymorphisms and adenocarcinoma. METHODS: This cross-sectional case control study was performed between Mach 2014 and January 2015 at the Tuba Hospital of Sari, Iran. Peripheral blood samples were collected in EDTA tube. DNA extraction was performed using the spin column procedure. The MT-2A polymorphisms MT-2A (rs1610216), (rs28366003) were determined by polymerase chain reaction-restriction fragment length polymorphism analysis in 95 a topic adenocarcinoma patients and 90 healthy individuals from Iranian population. RESULTS: The MT-2A rs1610216 polymorphism increased the risk of adeno carcinoma in our Iranian population [OR: 3.8533; 95%CI, 1.3155-11.2869; P=0.0139] and rs28366003 [OR: 4.0978; 95%CI, 1.2521-13.4108; P=0.0197]. CONCLUSION: The MT-2A gene polymorphism was associated with the risk of adenocarcinoma in the Iranian population.

RESUMO CONTEXTO: O câncer gástrico é a quarta causa mais comum de câncer em todo o mundo. Também em contraste com os enormes avanços na cura, a chance de viver é muito baixa, mesmo em casos de cirurgia. Ter uma predisposição genética desempenha um papel importante no desenvolvimento do câncer. A associação entre polimorfismos do gene metalotioneína-2A e o risco de adenocarcinoma tem sido amplamente estudada, mas há apenas um estudo sobre doenças estomacais. OBJETIVO: Neste estudo, objetivou-se investigar a associação entre 2 (MT-2A) polimorfismos e adenocarcinoma. MÉTODOS: Um estudo de controle de caso transversal foi realizado entre março de 2014 e janeiro de 2015 no hospital Tuba, Sari, Irã. Amostras de sangue periférico foram coletadas em tubo EDTA. A extração do ADN foi executada usando o procedimento da coluna da rotação. Os polimorfismos MT-2a MT-2A (rs1610216), (rs28366003) foram determinados pela análise do polimorfismo do comprimento do fragmento da reação-limitação de cadeia da polimerase em 95 pacientes com adenocarcinoma tópico e em 90 indivíduos saudáveis da população iraniana. RESULTADOS: O polimorfismo MT-2A rs1610216 aumentou o risco de adenocarcinoma de em nossa população iraniana. [OR: 3,8533; 95%CI, 1,3155-11,2869; P=0,0139] e rs28366003 [OR: 4,0978; 95%CI, 1,2521-13,4108; P=0,0197]. CONCLUSÃO: O polimorfismo do gene MT-2A foi associado ao risco de adenocarcinoma na população iraniana.

Molecular typing of Leishmania (Leishmania) amazonensis and species of the subgenus Viannia associated with cutaneous and mucosal leishmaniasis in Colombia: A concordance study / Tipificación molecular de Leishmania (Leishmania) amazonensis y especies del subgénero Viannia asociadas a la leishmaniasis cutánea y la mucosa en Colombia: un estudio de concordancia

Resumen Introduction: Multilocus enzyme electrophoresis (MLEE) is the reference standard for the characterization of Leishmania species. The test is restricted to specialized laboratories due to its technical complexity, cost, and time required to obtain results. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) is used to identify Leishmania species. Objective: To establish the concordance between the two tests as identifying methods for circulating species in Colombia. Materials and methods: A total of 96 isolates from patients with cutaneous or mucosal leishmaniasis were selected and identified by MLEE and PCR-RFLP with miniexon and hsp70 as the molecular targets, which were used sequentially. Restriction enzymes HaeIII and BccI were similarly applied. Cohen's kappa coefficient and the 95% confidence interval (CI) were calculated. Results: The kappa coefficient and the 95% CI between MLEE and PCR-RFLP displayed "very good" concordance with a coefficient of 0.98 (CI95%: 0.98 to 1.00). The identified species were Leishmania Viannia braziliensis, Leishmania Viannia panamensis, Leishmania Viannia guyanensis and Leishmania Leishmania amazonensis. A total of 80 of the 96 isolates were sequenced and the results obtained by PCR-RFLP were confirmed. Conclusion: Due to the concordance obtained between tests results with the amplification of the genes miniexon and hsp70, PCR-RFLP is proposed as an alternative for identifying circulating Leishmania species in Colombia.

Abstract Introducción. La electroforesis de enzimas multilocus (Multilocus Enzyme Electrophoresis, MLEE) es el estándar de referencia para la tipificación de las especies de Leishmania. La prueba está restringida a laboratorios especializados por su complejidad técnica, sus costos y el tiempo necesario para obtener resultados. La PCR-RFLP (Polymerase Chain Reaction-Restriction Fragment Length Polymorphism) se utiliza para tipificar especies de Leishmania. Objetivo. Establecer la concordancia entre las dos pruebas como métodos de tipificación de las especies circulantes en Colombia. Materiales y métodos. Se seleccionaron 96 aislamientos de pacientes con leishmaniasis cutánea o mucocutánea y se tipificaron mediante MLEE y PCR-RFLP con los blancos moleculares miniexon y hsp70 usados en serie. Las enzimas de restricción aplicadas fueron la HaeIII y la BccI, respectivamente. Se calculó el coeficiente kappa y un intervalo de confianza (IC) de 95 %. Resultados. Se determinó que la concordancia fue "muy buena" al obtener un coeficiente de 0,98 (IC95%: 0,98-1,00). Las especies identificadas fueron: Leishmania Viannia braziliensis, L. (V.) panamensis, L. (V.) guyanensis y L. (L,) amazonensis. De los 96 aislamientos, 80 se enviaron a secuenciación y se confirmaron los resultados obtenidos mediante PCR-RFLP. Conclusión. Dada la concordancia obtenida con la PCR-RFLP amplificando los genes miniexon y hsp70, se propone esta prueba como alternativa para la tipificación de especies de Leishmania circulantes en Colombia.

Identificación molecular de genotipos papilomavirus humanos en pacientes con cáncer de cuello uterino / Molecular identification of human papilloma virus genotypes in patients with cervical cancer

Fundamento: la principal causa para el cáncer cervico uterino es el papilomavirus humano de alto riesgo. No existen antecedentes de estudios moleculares para la tipificación de papilomavirus humano en la población de Camagüey. La reacción en cadena de la polimerasa es una técnica de Biología Molecular que se ha usado desde siempre para el diagnóstico clínico; esta permite confirmar la presencia del ADN del Papilomavirus en el ADN total extraído a partir de muestras de pacientes con cáncer de cuello uterino. Objetivo: demostrar por primera vez los genotipos papilomavirus humano de alto riesgo circulantes, que causan cáncer de cuello uterino en la población femenina de Camagüey, Cuba. Métodos: se realizó un estudio analítico prospectivo donde se estudiaron 22 pacientes femeninas de la provincia de Camagüey, que fueron atendidas en la consulta de Patología de cuello del Hospital Ginecoobstétrico. La identificación y tipificación de los genotipos papilomavirus humano se realizó mediante el procedimiento molecular polimorfismo de longitud en los fragmentos de restricción. Resultados: el 63, 6 % de los pacientes presentaron lesiones tipo exofítica, el 4, 5 % endofítica y el 31, 8 % de otros tipos. Este estudio confirmó que los genotipos papilomavirus humanos de alto riesgo circulantes en la provincia Camagüey son los genotipos 16 y 31, donde el más frecuente fue el genotipo 16. Conclusiones: la presente investigación constituye el primer reporte de un estudio molecular de papilomavirus humanos a partir de muestras de pacientes con cáncer de cuello uterino en la provincia de Camagüey, Cuba. Estos resultados, junto a los obtenidos por otros autores, tienen una contribución importante en el diseño de preparados vacunales preventivos o terapéuticos, cada vez más efectivo hacia una solución anticipada para el cáncer de cuello uterino en Cuba.

Background: it is demonstrated that the main cause of cervical cancer is high risk humanp virus. There is no precedent of molecular studies for the typing of Human Papilloma Virus in the population of Camagüey. Polymerase chain reaction is Molecular Biology technique that has been used traditionally for the clinical diagnosis and other purposes. This technique allows confirming the presence of papillomavirus´DNA in the total extracted DNA, from samples of patients with cervical cancer. Objective: to demonstrate for the first time existing high-risk human papilloma virus genotypes that cause cervical cancer in female population of Camagüey, Cuba. Methods: a prospective analytic study was conducted, in which 22 female patients of the province of Camagüey were studied. They received medical attention at Ana Betancourt Hospital. Identification and typing of the Human Papilloma Virus genotypes was carried through the molecular procedure Restriction Fragment Length Polymorphism. Results: patients who presented exophytic lesions accounted for 63, 6%, 4, 5 % had endophytic type, and 31, 8 % presented other types. This study confirmed that high-risk Human Papilloma Virus genotypes existing in the province of Camagüey are genotypes 16 and 31, and the most frequent is 16. Conclusions: this research is the first report of a molecular study of Human Papilloma Virus from samples of patients with cervical cancer in the province of Camagüey, Cuba. These results, along with the ones obtained by other authors, make an important contribution in the design of the increasingly effective therapeutic and preventive vaccine to an anticipated solution to cervical cancer in Cuba.

Lung Microbiome Analysis in Steroid-Naїve Asthma Patients by Using Whole Sputum / 결핵및호흡기질환

BACKGROUND: Although recent metagenomic approaches have characterized the distinguished microbial compositions in airways of asthmatics, these results did not reach a consensus due to the small sample size, non-standardization of specimens and medication status. We conducted a metagenomics approach by using terminal restriction fragment length polymorphism (T-RFLP) analysis of the induced whole sputum representing both the cellular and fluid phases in a relative large number of steroid naïve asthmatics. METHODS: Induced whole sputum samples obtained from 36 healthy subjects and 89 steroid-naїve asthma patients were analyzed through T-RFLP analysis. RESULTS: In contrast to previous reports about microbiota in the asthmatic airways, the diversity of microbial composition was not significantly different between the controls and asthma patients (p=0.937). In an analysis of similarities, the global R-value showed a statistically significant difference but a very low separation (0.148, p=0.002). The dissimilarity in the bacterial communities between groups was 28.74%, and operational taxonomic units (OTUs) contributing to this difference were as follows: OTU 789 (Lachnospiraceae), 517 (Comamonadaceae, Acetobacteraceae , and Chloroplast), 633 (Prevotella), 645 (Actinobacteria and Propionibacterium acnes), 607 (Lactobacillus buchneri, Lactobacillus otakiensis, Lactobacillus sunkii, and Rhodobacteraceae), and 661 (Acinetobacter, Pseudomonas, and Leptotrichiaceae), and they were significantly more prevalent in the sputum of asthma patients than in the sputum of the controls. CONCLUSION: Before starting anti-asthmatic treatment, the microbiota in the whole sputum of patients with asthma showed a marginal difference from the microbiota in the whole sputum of the controls.

Correlation between single nucleotide polymorphisms of multi-loci and multi-genes and myasthenia gravis / 中华神经科杂志

Objective To investigate the association between myasthenia gravis (MG) and single nucleotide polymorphisms (SNPs) of PTPN22 + 1858C/T,CTLA-4 (+ 49A/G;-1772C/T;-1661A/G),KRAS(rs9226),BCL2(rs4987855) and IGF-1R(rs34804698) genes.Methods Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was adopted to detect the gene types of SNPs in 76 MG patients who were enrolled in the Second Affiliated Hospital of Harbin Medical University from July 2011 to June 2015 and 59 healthy blood donors.Results In MG patients,the frequences of CTLA-4 +49A/G(rs231775) (57.9％) and-1772C/T (rs733618) (43.4％) were higher than that in the healthy controls (22.1％) (x2 =35.252,P =0.000; x2 =4.098,P =0.043).The frequence of CTLA-4 +49A/G in MG patients combined with thymoma (25.6％) was higher than other subgroups (thymic hyperplasia group:13.8％; normal thymus group:18.4％)(x2 =7.564,P=0.006; x2 =7.155,P=0.007).Meanwhile,the frequence of the C-1772 allele was higher in thymoma group (19.7％) compared with other two groups (thymic hyperplasia group:9.86％ ; normal thymus group:13.8％) (x2 =5.331,P =0.021 ;x2 =4.411,P =0.036).However,the other SNPs were not associated with the risk of MG.Conclusion There are associations of MG with CTLA-4 + 49A/G and-1772C/T SNPs,but not with PTPN,KRAS,BC12 and IGF-1R SNPs.

Correlation between the plasma level of MIF and its related gene-173G/C polymorphism and risk factors of atherosclerosis in pilots / 解放军医学杂志

Objective To investigate the relationship between the plasma level of macrophage migration inhibiting factor (MIF) and its related gene-173G/C polymorphism and risk factors of atherosclerosis in pilots for reducing the risk of adverse cardiovascular events in early stage. Methods Four hundred and fifty-eight military pilots undergoing medical examination (pilot group), 51 patients with coronary heart disease (CHD group), and 194 persons undergoing routine health examination (control group) were selected as the subjects under investigation. Subjects in pilot group were further grouped according to the different aircraft type they were flying and their flight time. General clinical data of the three groups were collected. ELISA was used to determine the plasma levels of MIF. MIF-173 G/C (rs755622) was detected by Taqman probe method. The differences of genotype and allele frequencies among the three groups were analyzed. Results No significant difference was found in plasma levels of MIF between pilot group and CHD group (P>0.05), but the levels were significantly higher in the both groups than in the control group (P0.05). There was no significant difference in genotype and allele frequencies among the three group (P>0.05). There was no significant difference of plasma MIF, TC, TG concentrations in the pilots who were with CC, GG and CG genotypes, respectively (P>0.05). Conclusions MIF-173G/C polymorphism may have no significant correlation to the early susceptibility of atherosclerosis. Elevated plasma MIF levels may be associated with the development of coronary heart disease.

Comparison of three methods for the genetic diagnosis of spinal muscular atrophy / 中华检验医学杂志

Objective To evaluate the value of PCR-restriction fragment length polymorphism (PCR-RFLP),real-time PCR and multiplex ligation-dependent probe amplification (MLPA) in the genetic diagnosis of spinal muscular atrophy (SMA) and make laboratory support accessible to clinicians for the molecular diagnosis of SMA.Methods Methodological evaluation.Forty-one suspected SMA cases and 359 control individuals received in Shanghai Children's Medical Centre from March 2013 to June 2014 were detected for the deletion of exon 7 and 8 in the survival motor neuron gene 1 (SMN 1) by PCR-RFLP,realtime PCR and MLPA,respectively.Then the results of the three methods were compared and the benefits and limitations of the three methods were evaluated.Results The result of real-time PCR was in complete agreement with that of MLPA,showing that 29 suspected cases harbored homozygous deletions of SMN1 and 1 case possessed heterozygous deletion.Among the homozygous deletions,27 patients demonstrated absence of exon 7 and 8,and 2 cases demonstrated only the absence of exon 7.Meanwhile,both PCR-RFLP and MLPA analysis showed the same results that only 5 out of 395 control cases carried heterozygous deletion.As for cases without heterozygous deletions,PCR-RFLP demonstrated the same result with real-time PCR and MLPA but it missed all the heterozygous ones.Conclusions PCR-RFLP,the conventional SMA gene diagnosis method,was only capable of detecting homozygous deletion of exon 7 and/or 8 of SMN1,but was not as sensitive as to find out the carriers with heterozygous deletions.MLPA could detect the deletion and quantify the copy numbers of exon 7 and 8 of SMN1,efficiently,while the price was relatively high,which brings challenges for its application in the carrier screening of SMA.Compared with these two methods,realtime PCR with high throughput and low input was a rapid,acourate and economic method for the genetic diagnosis of SMA and carrier screening in large populations.

MTHFR C677T polymorphism contributes to unexplained recurrent spontaneous abortion / 中华检验医学杂志

Objective To explore the relationship between methylenetetrahydrofolate reductase (MTHFR) 677C ＞ T and unexplained recurrent pregnancy loss (URPL).Methods All patients were recruited from the outpatient department of Obstetrics/Gynaecology & Genetics of Hangzhou First People's Hospital from January 2013 to May 2014.A case-control study was performed.According to the stochastic indicator method,there were 125 subjects with a history of ≥2 times URPL as the case group,and 905 healthy parous women with no history of URPL as the control group.Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to detect the distributions of the polymorphisms of MTHFR 677C ＞ T,and the results were validated using oligo-chip and direct sequencing.Results The allele and genotype frequencies of MTHFR were 60.0％ for C,40.0％ for T,32.0％ for CC,56.0％ for CT,and 12.0％ for TT in the case group and 67.4％ for C,32.6％ for T,41.4％ for CC,52.0％ for CT,and 6.6％ for TT in the control group,respectively.The prevalence of allele T was significantly higher in the case group than in the control group (OR =1.379,95％ CI =1.051-1.808,P =0.020),the frequency of genotype TT was also significantly higher in the case group than in the control group (OR =2.344,95％ CI =1.220-4.503,P =0.009).Conclusion The fertile women with MTHFR 677T allele and 677TT genotype may be susceptibility to URPL in a Chinese Han population from the Hangzhou area.

Polymorphisms of TNF-αgene and HSP70-2 gene in patients with acute pancreatitis / 天津医药

Objective To investigate the association of tumor necrosis factor (TNF)-α, heat shock protein (HSP)70-2 gene polymorphisms and susceptibility of acute pancreatitis(AP). Methods Using case-control method,The gene polymor?phism of TNF-α and HSP70-2 was detected by PCR-RLFP in 72 patients with AP and 71 healthy controls. Results There were no significant differences in proportion of TNF-αgenotype and alleles between AP and control groups (P>0.05). There were no significant differences in TNF-αgenotype and alleles between severe acute pancreatitis (SAP) and light acute pancreatitis (MAP) of AP group (P>0.05). There were no significant differences in white blood cell count, C-reactive pro?tein (CRP), amylase, three acyl glycerin and glucose between TNF-a and HSP70-2 gene of AA type and GA+GG type pa?tients (P>0.05). The HSP70-2 genotype GA+GG proportion was significantly higher in AP group than that of control group (69.4%vs 49.3%). The ratio of patients with G allele was significantly higher in AP group than that of control group(46.5%vs 31.7%). The ratio of patients with GA+GG type AP was significantly higher in SAP patients than that of MAP patients of AP group(81.0% vs 53.3%). There was no significant difference in G allele between SAP and MAP patients (P>0.05). Conclusion TNF-α polymorphisms is not associated with acute pancreatitis. There is an association between HSP70-2 polymorphisms and acute pancreatitis. Carrying the G allele increases the possibility of a severe acute pancreatitis ,which is one of the genetic susceptibility factors of severe acute pancreatitis.

Genetic diagnosis of 154 patients with suspected spinal muscular atrophy and its clinical value / 中华检验医学杂志

Objective To explore the clinical value of genetic diagnosis of SMA,the homozygous deletion of survival motor neuron 1 (SMN1) gene in suspected spinal muscular atrophy (SMA) patients were analyzed in this study.Methods A total of 154 patients suspected with SMA and 20 healthy volunteers were recruited from January 2007 to December 2014 in the Genetic Diagnosis Center of the First People's Hospital of Yunnan Province and the Department of Neurology of the Fourth Affiliated Hospital of Kunming Medical University.Potential deletions in exons 7 and 8 of SMN1 gene were screened by use of polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) method in both 154 patients suspected with SMA and 20 healthy volunteers.The frequencies of the deletions in exons 7 and 8 of SMN1 were calculated and statistical analysis of different deletion types among 3 SMA groups was performed with SPSS 13.0 software package.Results Among 154 suspected SMA patients,101 cases with homozygous deletions of exon 7 of SMN1 gene were detected,which accounted 65.6％ (101/154) of the suspected SMA patients.Among the 101 SMA patients,97.0％ (98/101) of the patients with both homozygous deletions of exons 7 and 8 for SMN1 gene and 3.0％ (3/101) of the patients with homozygous deletions of only exon 7 for SMN1 gene were detected.The patient with only deletion of exon 8 for SMN1 gene was notdetected.Four cases with negative results were subjected to be followed-up,but they were characteristic of SMA symptom by clinical re-visit.Thus,total 105 patients were confirmed with SMA,among them,68 were type Ⅰ SMA,27 were type Ⅱ SMA,and 10 were type Ⅲl SMA,which accounted for 64.8％ (68/105),25.7％ (27/105) and 9.5％ (10/105) of the SMA patients,respectively.Type Ⅳ SMA was not observed in these patients.No deletion was detected among 20 healthy volunteers.Conclusions PCR-RFLP assay is a noninvasive,simple,high sensitive and specific method for SMA diagnosis,which can be considered as the first-line genetic test for the suspected SMA patients.It will help to improve the accuracy of clinical diagnosis and the detection rate by strengthening the clinical diagnostic criteria and re-evaluating the suspected patients after negative genetic diagnosis.

Relationship of the interaction between age and gene polymorphisms with the susceptibility, invasion and metastasis of gastric carcinoma / 中华老年医学杂志

Objective To investigate the relationship of the interaction between age and polymorphisms of E-selectin gene A561C, chemokine receptor CCR2 gene A190G with the susceptibility, invasion and metastasis of gastric carcinoma.Methods Based on tumor-node-metastasis (TNM) staging classification, 750 patients with confirmed gastric carcinoma in our hospital from December 2011 to November 2014 were divided into 5 groups: stage Ⅰ, stage Ⅱ , stage Ⅲ, stage Ⅳ and stage 0 (n=150, each).No significant difference was observed in gender, ethnicity, birthplace and living habits among the 5 groups.Meanwhile, 750 healthy controls were selected in this study during the same time, and there was no significant difference in gender, ethnicity and birthplace between the healthy controls and patients with gastric carcinoma.The genetic polymorphisms of E-selectin gene A561C and chemokine receptor CCR2 gene A190G were analyzed by polymerase chain reaction-restriction fragment length polymorphisms (PCR-RFLP) in peripheral blood mononuclear cells (PBMs).Results The frequencies of CC (A561C) and GG (A190G) genotypes were 56.5％ and 56.8％ respectively in gastric carcinoma cases and 22.8％ and 23.1％ respectively in healthy controls, with statistically significant differences in the distribution frequencies between the two groups (P＜0.01 for all).The risk for gastric carcinoma significantly increased in subjects with CC (A561C) genotype (OR=4.4038, 95％CI=2.9421-7.2397) and in GG (190A/G) genotype (OR=4.3852, 95％ CI =2.8207-7.4942).Combined analysis of the polymorphisms showed that the distribution frequency of CC (A561C) genotype / GG (190A/G) genotype in gastric carcinoma cases and healthy controls was 46.4％ and 11.9％ respectively (P＜0.01).The positive interactions of age with CC (A561C) genotype and GG (190A/G) genotype for the risk of invasion and metastasis of gastric carcinoma were found (γ＞1 for both).The distribution frequencies of CC (A561C) genotype and GG (190A/G) genotype were 50.0％ and 50.0％ in stage Ⅰ , 63.4％ and 64.0％ in stage Ⅱ ,69.3％ and 69.3％ in stage Ⅲ, 76.7％ and 77.3％ in stage Ⅳ, and 23.3％ and 23.3％ in stage 0 respectively.Statistically significant differences were found in the distribution frequencies between stage 0 and the other 4 stages (P＜0.01 for all).The risks for the invasion and metastasis of gastric carcinoma were significantly increased in subjects with CC (A561C) genotype (ORⅠ-Ⅳ =3.2857-10.7959) and in those with GG (190A/G) genotype (ORⅠ-Ⅳ =3.2857-11.2101).Combined analysis of the polymorphisms showed that distribution frequency of CC (A561C) genotype / GG (190A/G) genotype had significant differences between the stage Ⅰ ～Ⅳ and stage 0 (39.3％, 53.3％, 59.3％,68.0％ vs.12.0％, P＜0.01).The proportion of elderly subjects were higher in Grade Ⅰ ～Ⅳ than in Grade 0 (51.3％, 62.7％, 70.0％, 75.3％ vs.26.7％, P＜0.01 for all).The risk for invasion and metastasis of gastric carcinoma was significantly increased in elderly patients (ORⅠ-Ⅳ =2.9001 ～8.3986).The positive interactions of age with CC (A561C) genotype and GG (190A/G) genotype for the risk of invasion and metastasis of gastric carcinoma were found (γ＞ 1 for All).Conclusions Age and E-selectin gene A561C (CC) and chemokine receptor CCR2 gene A190G (GG) are the risk factors for the invasion and metastasis of gastric carcinoma, and the interactions between age and genetic polymorphisms increase the risk of invasion and metastasis of gastric carcinoma.

Association of ERα-29 gene polymorphisms with susceptibility to HBV-related hepatocellular carcinoma / 临床肝胆病杂志

ObjectiveTo evaluate the relationship between estrogen receptor-α-29 (ERα-29) gene polymorphisms and the development of HBV-related hepatocellular carcinoma (HCC) in Gansu Province, China, and to investigate the pathogenesis of HCC at the gene level. MethodsGene polymorphisms of ERα-29 were analyzed in 106 HBV-related HCC patients and 98 healthy individuals as normal controls using the polymerase chain reaction-restriction fragment length polymorphism technique. Population allele frequencies were calculated using the gene counting method and then tested using the Hardy-Weinberg law of genetic equilibrium. Comparisons of genotype and allele frequencies between groups were performed using the χ2 test. ResultsThe frequencies of TT genotype and T allele of ERα-29 gene in HBV-related HCC patients were significantly higher than those in the normal controls, i.e., 31.1% and 53.8% vs. 11.2% and 32.1% (χ2 = 3.449, P＜0.05; χ2 = 3.840, P＜0.05). In contrast, the frequencies of CC genotype and C allele of ERα-29 gene in HBV-related HCC patients were significantly lower than those in the normal controls, i.e., 23.6% and 46.2% vs. 47.0% and 67.9% (χ2 = 3.488, P＜0.05; χ2 = 3.840, P＜0.05). Compared with those carrying C allele, carriers of T allele had an increased risk (2.46-fold) of HBV-related HCC (OR = 2.46, 95% CI: 1.64-3.69). Conclusion T allele of ERα-29 gene can increase the risk of HBV-related HCC.

The relationship between human adiponectin gene polymorphism of Guangxi Zhuang ethnic group and gestational diabetes mellitus / 中国医师杂志

Objective To investigate the association of single nuchotide polymorphism (SNP) + 45 T/G and + 276 G/T in the adiponectin gene with type 2 diabetes in Guangxi Zhuang pregnant women.Methods Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method was used to determine the distributions of allele and genotype frequencies of SNP + 45T/G polymorphism and SNP + 276 G/T polymorphism in adiponect in gene in 98 type 2 diabetic patients and 135 normal control subjects.Results The TG,GG genotype frequencies of the SNP +45T/G were higher in gestational diabetes mellitus (GDM) than those in controls (P ＜ 0.05).The G-allele frequency was significantly higher than those in controls (P ＜ 0.05).The distribution of genotype and allele frequencies of the SNP + 276 G/T polymorphism was not different in diabetes and control groups (P ＞ 0.05).Women carrying the TG and GG allele of SNP +45T/G showed higher TG (P ＜0.05),and lower APN levels than those with TT genotype (P ＜0.05).Conclusions G allele of SNP +45T/G was related to GDM possibly.T→G variation might involve in the occurrence of GDM by reducing serum diponectin levels.Monitoring this site in pregnant women could predict the risk of GDM.

Relationship between monocyte chemoattractant protein-1 gene polymorphism and susceptibility of epithelial ovarian cancer / 中国医师杂志

Objective To investigate the relationship between monocyte chemoattractant protein-1 (MCP-1) A-2518G single nucleotide polymorphism (SNPs) and susceptibility of epithelial ovarian cancer(EOC) in Chinese Han population of Hunan region.Methods MCP-1 A-2518G SNPs of the EOC were genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis in 92 patients with EOC and 38 healthy women as control.Results MCP-1 A-2518G SNPs had AA,AG,and GG genotypes in cancer and control groups.The frequencies of AA,AG,and GG genotypes were not significantly different between cancer and control groups (AA:17.40％ and 15.79％ ; AG:44.56％ and 52.63％ ; and GG:38.04％ and 31.58％ ; P ＞0.05).Multivariate logistic regression analysis showed that there was no significant correlation between MCP-1 A-2518G polymorphism and EOC (P ＞0.05).Conclusions This present study suggested that MCP-1 A-2518G polymorphism should not be related to susceptibility of EOC in the Chinese Han population of Hunan region.

Sequencing Technology in Molecular Diagnosis of Spinal Muscular Atrophy Caused by SMN1 Deletion / 天津医药

Objective To investigate the feasibility of DNA sequencing analysis in molecular diagnosis for spinal muscular atrophy (SMA). Methods Two pairs of primers were utilized to amplify the region including 5 different bases in SMA-causative gene SMN1 and its homologue copy SMN2 by polymerase chain reaction (PCR). The first primer amplified a fragment 501 bp long spanning from SMN intron 6 to intron 7 targeting four different bases (g.31957, 32006, 32154 and 32269). The second primer reversely amplified a 189 bp long fragment within SMN exon 8 including one base-pair differ-ence (g.32734). PCR procedure was followed by Sanger sequencing technique to identify the 5 different bases. SMA patients caused by SMN1 homozygous deletion were distinguished from carriers or normal controls by absence of SMN1 specific bas-es in sequence chromatograms. This assay was performed in 7 SMA suspected patients and their parents. The specimens were also detected by PCR- restriction fragment length polymorphism (RFLP) method. Results It was found that 6 of 7 SMA suspected patients showed only SMN2 specific bases at the 5 different base positions among the region from intron 6 to exon 8, which meant the patient displaying only SMN2-specific nucleotide a, T, g, g and A at g.31957, 32006, 32154, 32269 and 32734, while their parents (carriers) showed a/g, T/C, g/a, g/a and A/G at the same sites. SMN1 gene was deleted in the patient, and the deletion region was inferred from intron 6 to exon 8. Because carriers had both SMN1 and SMN2 genes, they can be discriminated from the SMN1 deleted patient. One of 7 patients yield an unique sequence chromatogram of a, T, g, g and A/G, indicating that exon 8 of SMN1 was not deleted in this patient. Conclusion DNA sequencing analysis is an alter-native simple method for detecting SMA caused by homozygous deletion of SMN1. We recommend to replace the widely used PCR-RFLP method with DNA sequencing assay.

Determination of gene polymorphism and serum concentration of mannose-binding lectin in patients with psoriasis / 中华皮肤科杂志

Objective To determine the gene polymorphism and serum concentration of mannose-binding lectin (MBL) in patients with psoriasis,and to analyze the relationship between MBL and psoriasis.Methods Totally,67 patients with psoriasis vulgaris and 69 healthy human controls were enrolled in this study.Venous blood samples were obtained from all the subjects.Genomic DNA was extracted,and PCR-restriction fragment length polymorphism (PCR-RELP) analysis was conducted to determine the polymorphism at codon 54 of the MBL gene.Enzyme-linked immunosorbent assay was performed to measure the serum level of MBL.A chi-square goodness-of-fit test was carried out to evaluate Hardy-Weinberg equilibrium,t test to compare the serum concentration of MBL,and chi-square test to compare the frequency of genotypes and alleles of MBL gene codon 54.Results The patients with psoriasis showed higher frequency of GGC/GAC heterozygote but lower frequency of GGC/GGC homozygote (x2 =10.36,P ＜ 0.05),together with increased frequency of GAC allele but decreased frequency of GGC allele (x2 =8.31,P ＜ 0.05),at codon 54 of the MBL gene compared with the healthy controls.The variant allele GAC at codon 54 of the MBL gene was markedly associated with psoriasis (OR =3.383,95％ CI 1.585-7.211,P ＜ 0.05).The serum concentration of MBL was (2.193 7 ± 0.816 3) mg/L in patients with psoriasis,significantly lower than that in the healthy controls ((3.269 5±1.205 8) mg/L,t=6.11,P＜ 0.05).Conclusion MBL might be associated with the pathogenesis of psoriasis to some degree.