Establishment and verification of a real-time quantitative PCR method for determination of exogenous antibody gene copy number in CHO cells / 中国生物制品学杂志

@#Objective To establish a real-time quantitative PCR method using SYBR GreenⅠto detect the copy numbers of light chain(LC)and heavy chain(HC)of exogenous antibody gene in CHO cells,and verify and preliminarily apply this method.Methods With the B2m(β2-microglobulin)expressed stably in CHO cells as the internal reference gene,suitable primers of LC,HC genes and internal reference gene were designed respectively,and the reaction system and program of the real-time quantitative PCR method were determined. The established method was verified for the specificity,linearity,precision and durability,and used to detect the copy numbers of LC and HC genes in the recombinant cell lines of working cell bank(WCB)and cells of different passages.Results The primers of exogenous genes and internal reference gene showed specific binding to the target fragments;The efficiency of primer amplification for the B2m gene,LC gene,and HC gene was 106. 7%,106. 3% and 99. 1%,respectively,and the correlation coefficients of the linear equations were all greater than 0. 99 with a good linear relationship;The relative standard deviations(RSDs)of precision verification were all less than 1%;Few cycles of freeze-thaw in a short period had little effect on the detection results. The copy numbers of LC and HC genes in different generations of recombinant cell lines detected by the established method showed no obvious changes.Conclusion A real-time quantitative PCR method for the determination of the copy number of exogenous genes in CHO cells was successfully established with good specificity,linearity,precision and durability,which provides a reference for detecting the copy number of exogenous genes expressed in other CHO cell lines

Isolation of Enterobacteriaceae strains carrying mcr-1 resistance gene from Shanghai wastewater treatment plants and quantification of their copy number / 上海预防医学

ObjectiveTo provide technical support for the molecular surveillance of pathogenic bacteria strains carrying mobile colistin resistance-1 （mcr⁃1） gene isolate from inlet of wastewater treatment plants （WWTP）. MethodsThe Enterobacteriaceae strains carrying mcr⁃1 resistance gene isolate from inlet of WWTP during April 1 to June 30， 2023 in Shanghai were cultured on blood-rich and SS culture medium and were identified using a mass spectrometry analyzer. The mcr⁃1 gene and copy number were detected by real-time fluorescence quantitative PCR. Drug susceptibility test was performed by microbroth dilution method. The copy numbers of Escherichia coli carrying mcr⁃1 gene isolated from wastewater and human fecel were statistically analyzed by SPSS 25.0. ResultsA total of 14 strains carrying the mcr⁃1 gene were isolated from 49 WWTP samples， and the positive isolation rate was 28.6%， including 12 non-diarrheal E. coli strains and 2 Klebsiella pneumoniae strains. The drug susceptibility results showed that all 14 strains were multi-drug resistant bacteria. They were all sensitive to imipenem and tigecycline， but were ampicillin- and cefazolin-resistant. There was no significant difference in the copy number between human-sourced diarrheal E. coli and wastewater-sourced non-diarrheal E. coli （t=0.647， P>0.05）. ConclusionThe isolation and identification of strains carrying the mcr⁃1 gene from inlet of WWTP samples were firstly established in Shanghai. The multi-drug resistance among the isolated strains is severe. To effectively prevent and control the spread of colistin-resistant bacteria， more attention should be paid to the surveillance of mcr⁃1 gene.

Analysis of cell mutation types of colorectal neuroendocrine tumors / 基础医学与临床

Objective To investigate the mutation types of colorectal neuroendocrine tumors(NETs)and better un-derstand the pathogenesis of colorectal nets.Methods Patients undergoing colorectal NETs surgery were recruited,colorectal NETs and corresponding adjacent cancerous tissues were collected,and whole genome sequencing(WGS)was performed and further deeply analyzed.Results WGS sequencing showed that the mutation types of colorectal NETs included single nucleotide mutations,insertion and deletion mutations(InDel,less than 50 bp in length),copy number variations(CNV),and large structural variations(SV,more than 50 bp in length),such as insertion(INS),deletion(DEL),intra chromosomal translocation(ITX),inter chromosomal translocation(CTX)and inversion(INV).Conclusions A large number of somatic mutations occur in colorectal NETs,especially chro-mosome translocation

Factors influencing number of lymph nodes obtained by prophylactic central lymph node dissection for pap-illary thyroid cancer / 实用医学杂志

Objective To investigate the factors affecting the number of lymph nodes obtained by unilateral lobectomy and isthmus combined with prophylactic ipsilateral central lymph node dissection for papillary thyroid cancer,and to analyze the pattern of change so as to provide an important basis for clinical treatment of papillary thyroid cancer.Methods Retrospective analysis was performed on the clinical and pathological data of 193 patients admitted to the department of Thyroid Surgery of the First Affiliated Hospital of Jinzhou Medical University from January 2019 to January 2022 for papillary thyroid cancer who underwent unilateral glandular lobe and isthmus resec-tion combined with prophylactic ipsilateral central lymph node dissection.The number of obtained lymph nodes was divided into a group with high number of obtained lymph nodes(n>6)and a group with low number of obtained lymph nodes(n≤6).Univariate analysis and multivariate analysis were used to explore the factors affecting the num-ber of obtained lymph nodes.The influence of the number of lymph nodes on the central lymph node metastasis was evaluated by correlation analysis,and the influence of the number of lymph nodes on the quality of life of patients by postoperative complications analysis.Results The number of lymph nodes obtained was positively correlated with the number of lymph node metastases in the central region(r = 0.240,P<0.05)and the rate of lymph node metastasis(r = 0.161,P<0.05),respectively.The number of lymph node metastases controlled remained unchanged,and the rate of lymph node metastasis decreased with the increase of the number of obtained lymph nodes(r =-0.444,P<0.05).Univariate analysis of the number of lymph nodes between the two groups showed no significant differences in terms of sex,age,operation duration,BMI,maximum tumor diameter,multifocality,Hashimoto's thyroiditis,focal location,capsular invasion,contralateral thyroid nodule,parathyroid transplantation and BRAF gene V600E mutation(P>0.05).There was statistical significance in the application of nano carbon between the two groups(P<0.05).Logistic binary regression analysis showed that the combination of Hashimoto's thyroiditis and the application of carbon nanoparticles were independent influencing factors for the high number of lymph nodes obtained(P<0.05).ROC curve analysis suggested that the area under the curve occupied by carbon nanoscale was 0.658(95%CI:0.580～0.735,P<0.05),and the area under the curve occupied by Hashimoto's thyroiditis was 0.584(95%CI:0.504～0.665,P<0.05).In addition,there was no significant difference in the incidence of vocal cord paralysis and lymphatic leakage between the two groups(P>0.05).Conclusions When the number of lymph nodes in the cen-tral region remained unchanged,the higher the number of lymph nodes obtained,the lower the rate of lymph node metastasis.The application of nano-carbon and patients with Hashimoto's thyroiditis could increase the number of lymph nodes obtained,and there is no significant difference in the incidence of postoperative complications between high and low number of lymph nodes obtained.

Effect of vessel number on computational fluid dynamics in vascular networks / 中国组织工程研究

BACKGROUND:Mechanical factors can affect the angiogenic ability of vascular endothelial cells.How the vessel number affects the hydrodynamic properties of microvessels remains to be clarified. OBJECTIVE:To investigate the influence of vessel number on the hydrodynamics of vascular networks based on computational fluid dynamics. METHODS:Three three-dimensional models of vascular network with different vessel numbers were constructed using the Geometry module of ANSYS 19.0 software,and then the vascular network was meshed to tetrahedral elements in Mesh module.The vascular network was assumed to rigid wall without slip,and the blood was assumed to laminar,viscous,and incompressible Newtonian fluid.Blood density,velocity,and a series of blood viscosity coefficients were also established.The Navier-Stokes equation was used for calculation.Hydrodynamic properties of different parts of vascular network with different vessel numbers were analyzed and compared. RESULTS AND CONCLUSION:The streamline,velocity,and mass flow all had the same trend in the vascular network,that is,the outlet and inlet were higher and the middle junction of vascular network was lower.The more the number of vessels,the thinner the blood flow lines in each part of the vascular network.Also,the velocity,mass flow,and wall shear decreased with the increase of the number of blood vessels.Therefore,the changes in vessel number could influence the hydrodynamic environment in the vascular network.Computational fluid dynamics indicates that the changes in vessel numbers can influence the hydrodynamic properties of blood,and provides a new idea for treating bone hypoperfusion-induced diseases(fracture nonunion,bone defect,osteoporosis,etc.)through tonifying kidney and activating blood circulation based on the coupling between angiogenesis and osteogenesis.

Study on the value of dual energy index and atomic number combined with slope of energy spectrum curve in identifying components of urinary tract stones based on dual energy CT / 中国医学装备

Objective:To assess a value of dual energy index(DEI)and effective atomic number(Zeff)of dual-energy computed tomography(DECT)combined with slope of energy spectrum curve in identifying the component of urinary tract stones.Methods:The clinical and DECT imaging data of 111 patients with urinary tract stones who admitted to Nantong Haimen People's Hospital from October 2019 to October 2022 were selected,and the diameters of the urinary tract stones of all patients were<5 mm.The components of outside body stones of all patients were analyzed by infrared spectrum.The accuracy,sensitivity and specificity of the DEI,Zeff,slope of energy spectrum curve and the combination of the them were respectively calculated in identifying the components of urinary tract stones.Results:In 111 patients,the patients with calcium oxalate stones were 75(67.57%),and ones with hydroxyapatite were 15 cases(13.51%),and ones with uric acid stones were 21 cases(18.92%),and there were no other type of stone included cysteine.The differences of CT values(F=487.691,P<0.001),DEI values(F=395.553,P<0.001),Zeff values(F=818.689,P<0.001)and the slopes of energy spectrum curves(H=19.615,P<0.001)were statistically significant among the three types of stones.The overall accuracies of DEI,Zeff and slope of energy spectrum curve were 87.39%(97/111),84.68%%(94/111)and 81.98%(91/111)in identifying the components of urinary tract stone.The range of diagnostic accuracies of DEI,Zeff and the slope of energy spectrum curve was between 80%and 100%for calcium oxalate stones,hydroxyapatite and uric acid stones.The overall accuracy of DEI,Zeff combined with slope of energy spectrum curve was 94.59%(105/111)in identifying the components of urinary tract stones.The diagnostic accuracies of DEI,Zeff combined with slope of energy spectrum curve were respectively 94.59%,94.59 and 100%for calcium oxalate stones,hydroxyapatite and uric acid stones.Conclusion:Compared with the single indicator of DECT,the DEI and Zeff combined with the slope of the energy spectrum curve showed better diagnostic accuracy in identifying the components urinary track stones.

Analysis on the Factors Influencing the Human Resource Allocation in Tertiary Public Traditional Chinese Medicine Hospitals / 中国医院管理

Objective Starting from the actual numbers of health personnel of tertiary public hospitals of Traditional Chinese Medicine(TCM),to quantitatively analyze the influencing factors on the allocation of human resources and obtain a prediction model.Methods The balanced panel data from 517 Tertiary Public TCM Hospitals in the period of 2011-2020 were collected,and the two-way fixed effects model was used to empirically analyze the impact of scale,demand and other factors on the actual number of health personnel in these hospitals.Result The number of beds is a key factor affecting the human resource allocation of Public TCM Hospitals,and various factors such as de-mand,policy,price,efficiency,and administrative management also have significant impacts on the allocation.The demand for outpatient services,government financial support,and efficiency of resource utilization are all promoting factors,while the increase in human resource prices,income generation efficiency,and administrative manage-ment levels have negative effects.A prediction model is proposed.Conclusion The planning principle of matching bed numbers with human resources allocation is in line with the actual environment.When predicting the total personnel allocation or authorized strength,various factors should also be fully considered,which can provide reference for the formulation of human resource policies in Public TCM Hospitals.

Role of echocardiography in assessing copy number variation in fetal congenital heart disease / 中华围产医学杂志

Objective:This study examines the application of echocardiography in the prenatal diagnosis of copy number variation (CNV) associated with fetal congenital heart disease (CHD).Methods:A retrospective analysis was conducted on 447 singleton pregnancies from Quanzhou Maternal and Child Care Hospital (Quanzhou Children's Hospital) from January 2019 to August 2022. These individuals underwent echocardiographic assessments suggestive of fetal CHD and subsequently received invasive prenatal diagnoses. Comprehensive karyotype analysis and chromosome microarray analysis (CMA) were performed for each case. The discrepancies in the chromosomal abnormality detection were analyzed between the results produced by CMA and karyotype analysis. Furthermore, differences in the detection of pathogenic copy number variation (pCNV) between the two methods in CHD cases with diverse cardiac phenotypes, including the presence or absence of extracardiac structural malformations, the type, and quantity of cardiac structural anomalies, were explored. Statistical analysis was conducted using the Chi-square test. Results:Compared with conventional karyotype analysis, CMA demonstrated a higher detection rate of fetal chromosomal abnormalities [10.5% (47/447) vs. 20.6% (92/447), χ 2=161.56, P<0.001]. In terms of distinct cardiac phenotypes, CHD cases with extracardiac structural anomalies displayed an escalated pCNV detection rate in comparison to isolated CHD cases [11.4% (45/394) vs. 32.1% (17/53), χ 2=16.68, P<0.001]. Within the cardiac structural anomaly subgroups, increased pCNV detection rates were observed in the septal defect subgroup, conotruncal malformation subgroup, and left ventricular malformation subgroup [18.4%(29/158), 25.9%(7/27), and 25.0%(7/28) vs. 7.6%(16/210); χ 2=9.15, 9.68, and 8.55, respectively, all P<0.05]. The CMA-identified pCNV correlated with CHD included 22q11.2 deletions/duplications in eight cases, 4p16.3 deletions in two cases, 11q23.3 microduplications in two cases, 1q21.1 microdeletions/microduplications in two cases, 4q28.3 microduplications in one case, and 10p15.3 microdeletions in one case. Conclusions:CMA technology exhibited an enhanced ability to detect pCNV in fetuses with CHD. Echocardiography can guide targeted CMA screening, thereby facilitating prenatal genetic assessment of CHD.

Measurement of the relationship between maxillary premolar roots and the maxillary sinus floor using cone beam CT and analysis of the impact on immediate implantation / 口腔疾病防治

Objective@#To analyze the spatial relationship between the roots of maxillary anterior premolars and the maxillary sinus, thus providing an anatomical basis for timing, planning, surgical approaches, and implant selection at this site.@*Methods@#Cone beam CT (CBCT) images were collected from 264 patients (aged 20-65 years) who visited the Ruihua Dental Clinic between January 2017 and March 2023. The minimum distance from the apex of the maxillary anterior premolar roots to the lower wall of the maxillary sinus was measured on the coronal plane. The classification of the vertical relationship between the tooth root and the lower wall of the maxillary sinus was performed, and comparisons were made bilaterally, between genders, and among different age groups.@*Results@#The minimum distance (Q50) from the apex of the first maxillary premolar root to the lower wall of the maxillary sinus was 7.34 mm for the single-root type, 7.80 mm for the buccal root of the double-root type, and 7.36 mm for the palatal root. For the second maxillary premolar, the median distance was 2.56 mm for the single root type, 1.73 mm for the buccal root type, and 1.23 mm for the palatal root type. There was a significant difference in the shortest distance from the apex of the right second maxillary premolar single root to the lower wall of the maxillary sinus among the different age groups (P<0.05), with the 20-29-year-old group having the smallest median distance (1.52 mm) and the ≥ 40-year-old group having the largest (4.44 mm). There was no significant difference in the effect of sex or laterality on distance (P>0.05). The most common vertical relationship between the apex of the maxillary anterior premolar roots and the lower wall of the maxillary sinus was noncontact. There was no significant difference in the vertical relationship classification between the single-root and double-root types (P>0.05).@*Conclusion@#Most maxillary first premolar roots can provide sufficient bone height, which makes it easy to achieve immediate implantation. The maxillary second premolar root frequently involves insufficient bone, which is necessary to make full use of the bone wall of the extraction socket or the sinus floor cortical bone to achieve initial stability. The vertical relationship between the premolar root and maxillary sinus was influenced by age and dental position. Younger age groups often exhibit inadequate bone height, and the indication for immediate implantation should be carefully considered. The number of roots does not significantly affect the relationship between the sinus and root; however, double-rooted premolars offer more support for immediate implantation and socket healing due to the small root diameter and bony separation between the roots.

Ribosomal DNA copy number variation in peripheral blood and its influencing factors among patients with pneumoconiosis / 预防医学

Objective@#To explore the changes in ribosomal DNA copy number in peripheral blood among patients with pneumoconiosis and its influencing factors, so as to provide insights into prevention and treatment of pneumoconiosis.@*Methods@#Eighty-eight patients with pneumoconiosis who visited a designated hospital and 71 community residents with no history of pneumoconiosis or dust exposure were selected as the pneumoconiosis group and control group, and age, smoking history, drinking history and cumulative years of exposure to dust were collected through questionnaire surveys. The copy number of 45S rDNA and 5S rDNA was detected using real-time fluorescence quantitative PCR, and the differences between the two groups were compared. Factors affecting the copy number of 45S rDNA and 5S rDNA were identified by a multiple linear regression model.@*Results@#The pneumoconiosis group had a median age of 56.00 (interquartile range, 15.25) and a mean cumulative dust exposure duration of (12.40±8.08) years, with 56.82% smoking and 62.50% drinking. The control group had a median age of 64.00 (interquartile range, 37.00) years, with 32.39% smoking and 26.76% drinking. The median copy number of 45S rDNA in the pneumoconiosis group was 1.29 (interquartile range, 0.59), which was lower than 2.10 (interquartile range, 1.88) in the control group; the median copy number of 5S rDNA in the pneumoconiosis group was 5.33 (interquartile range, 0.85), which was higher than 4.66 (1.34) in the control group (both P<0.05). Multiple linear regression analysis identified age (β=-0.034) and pneumoconiosis (β=-1.595) as factors affecting 45S rDNA copy number, age (β=-0.013) as a factor affecting 5S rDNA copy number, and age (β=0.018) as a factor affecting 5S rDNA copy number in the pneumoconiosis group (all P<0.05).@*Conclusions@#Compared with community residents with no history of pneumoconiosis or dust exposure, the copy number of 45S rDNA in peripheral blood among patients with pneumoconiosis is reduced and the copy number of 5S rDNA is increased.

Effect of reduced daily magnesium oxide doses on laxative effect: a single-center retrospective study / Journal of Rural Medicine

Objective: To investigate the laxative effect of reducing the number of daily doses of magnesium oxide (MgO), while maintaining the total daily dose of MgO in patients with good bowel movements.Patients and Methods: The retrospective analysis involved 11 patients with regular bowel movements who were prescribed MgO for constipation upon admission to a nursing care facility accompanied by home visits by a pharmacist. This investigation was conducted before and after reducing the number of daily doses from three to two, or from two to one, over a two-week period.Results: The number of bowel movements was 7.6 ± 3.4 and 6.6 ± 4.0 times for two weeks before and after the change in dosage frequency, respectively. The difference was not statistically significant (P=0.09). The Bristol Stool Form Scale was 3.9 ± 0.9 and 4.0 ± 0.9 two weeks before and after the change, respectively, which was not significant (P=0.93). Two weeks after the change, the MgO regimen remained unchanged and no on-demand laxatives were administered.Conclusions: The results suggest that reducing the number of daily doses of MgO does not affect its laxative action.

Application effect analysis of a horizontal endoscope storage cabinet in storage of flexible endoscopes / 中国内镜杂志

Objective To explore the influence of different storage methods on flexible endoscope.Methods 500 compliant soft endoscopes in the endoscopy center of our hospital from January 2019 to January 2020 were selected as research objects.The endoscopes of the observation group were placed horizontally in the storage cabinet,while those of the control group were placed vertically in the storage cabinet.The effects of the two storage methods on bacterial colony number,quality qualified rate and quality of soft endoscopes were compared.Results The study found that the number of bacterial colonies in the flexible endoscope in the 2 h storage of the observation group was(10.27±2.22)cfu/piece and the control group was(13.18±1.33)cfu/piece,and the difference was not statistically significant(P>0.05);The number of bacterial colonies in the flexible endoscope in the 72 h of the observation group was(14.75±2.00)cfu/piece,and the control group was(223.28±17.07)cfu/piece,and the difference was statistically significant(P<0.05).The observed cost of consumables and labor were obliviously lower than those of the control group,there was statistical significance(P<0.05).Conclusion The flexible endoscope is placed horizontally in the designated storage cabinet,which can eliminate the daily morning pre-cleaning within 72 h,and will not increase the damage rate of endoscope.It is a clinically recommended storage method for endoscopes.

Factors affecting poor sleep quality in last trimester pregnant women: a cross-sectional research from Turkey

SUMMARY OBJECTIVE: The aim of the study was to determine the factors affecting poor sleep quality in the last trimester pregnant women. METHODS: A cross-sectional study was conducted at a tertiary care maternity hospital in Ankara, Turkey. The research was conducted between May and September 2019 with 570 pregnant women in the last trimester. The data were collected through the Personal Information Form, Pittsburgh Sleep Quality Index, International Physical Activity Questionnaire Short Form, Restless Legs Syndrome Form, Brief Fatigue Inventory, and Perceived Stress Scale. RESULTS: The mean Pittsburgh Sleep Quality Index score of the pregnant women was 5.98±3.31, and 48.9% of them were found to have over five Pittsburgh Sleep Quality Index scores. Hemoglobin levels, income perceptions, smoking habits, attending pregnant schools, experiencing leg pains or cramping, experiencing back, waist, or neck pains, Restless Legs Syndrome, fatigue levels, and perceived stress levels of the pregnant women were found to be important determinants of sleep quality (p<0.05). CONCLUSION: According to the findings, increasing hemogram levels, attending antenatal education programs, and improving the ability of pregnant women to manage stress are opportunities to improve sleep quality during pregnancy. Careful evaluation of pregnant women in terms of insomnia and affecting factors can be suggested during antenatal follow-up.

Associação entre número de dentes, próteses dentárias e disfagia autorreferida em idosos brasileiros: estudo populacional / Association between number of teeth, dental prostheses, and self-reported dysphagia in brazilian old people: a population-based study

RESUMO Objetivo Verificar a associação entre o número de dentes e uso de prótese dentária removível e a ocorrência de disfagia autorreferida em idosos de 60 anos ou mais. Método Estudo transversal de base populacional com 5.432 idosos, que participaram da linha de base do Estudo Longitudinal da Saúde do Idoso (ELSI-Brasil). O desfecho "disfagia" foi associado ao número de dentes permanentes e ao uso de prótese dentária removível. As variáveis independentes sociodemográficas (idade, sexo e cor/raça) e de histórico clínico (nenhuma morbidade, uma morbidade ou mais de duas morbidades) utilizando Regressão de Poisson com variância robusta e seus respectivos intervalos de confiança (IC) de 95% foram analisados. Resultados A prevalência de disfagia autorreferida nos idosos não institucionalizados foi de 30%. O grupo de idosos com 10 - 19 dentes permanentes apresentou um risco de 52% de queixa de disfagia autorreferida (RPaj 1,565 IC95% 1,34;1,826) se comparado com seus pares com mais dentes. Conclusão foi encontrada associação entre o menor número de dentes e próteses removíveis com a ocorrência de disfagia.

ABSTRACT Purpose To investigate the association between the number of permanent teeth and the use of removable dental prostheses with self-reported dysphagia occurrence in individuals aged 60 years or older. Methods A population-based cross-sectional study was conducted with 5,432 old individuals who participated in the baseline of the Brazilian Longitudinal Study of Elderly Health (ELSI-Brazil). The outcome "dysphagia" was associated with the number of permanent teeth and the use of removable dental prostheses. Sociodemographic independent variables (age, sex, and race/ethnicity) and clinical history variables (no morbidity, one morbidity, or more than two morbidities) were analyzed using Poisson Regression with robust variance and their respective 95% confidence intervals (CI). Results The prevalence of self-reported dysphagia in non-institutionalized old individuals was 30%. The group of old individuals with 10 - 19 natural teeth showed a 52% increased risk of self-reported dysphagia complaint (PRadj 1,565 IC95% 1,34;1,826) compared to their counterparts with more teeth. Conclusion An association was found between a lower number of teeth and removable prostheses with the occurrence of dysphagia.

Amplificación de sondas múltiples dependientes de ligación: actualización y revisión del principio, variantes y aplicaciones / Multiplex ligation-dependent probe amplification: update and review of principles, variants, and applications / Amplificação de múltiplas sondas dependentes de ligação: atualização e revisão do princípio, variantes e aplicações

Resumen La amplificación de sondas múltiples dependientes de ligación (MLPA) es una valiosa herramienta en el estudio de alteraciones en el número de copias para distintas patologías de origen genético. La existencia de una amplia oferta de kits comerciales, el fácil y rápido procesamiento de laboratorio, su alta sensibilidad y, en general, los buenos resultados informados han permitido que su uso se encuentre expandido en muchos laboratorios de biología molecular alrededor del mundo. El principio de esta técnica ha sido adaptado para distintas aplicaciones como la MLPA metilación específica para el estudio de enfermedades relacionadas con la impronta epigenética y la MLPA digital que se acopla a equipos de secuenciación de nueva generación para aumentar la cantidad de regiones analizadas en un solo ensayo. Al contar con 10 años de experiencia en el uso de esta técnica en el Laboratorio Nacional de Tamizaje Neonatal y Alto Riesgo se realiza esta revisión con el fin de analizar los principios, variantes de la técnica, análisis de los datos, algunas aplicaciones, ventajas y desventajas de la MLPA en comparación con otras tecnologías disponibles.

Abstract Multiplex ligation-dependent probe amplification (MLPA) is a valuable tool in the study of copy number alterations for different pathologies of genetic origin. The availability of a wide range of commercial kits, the easy and fast laboratory processing, its high sensitivity and, in general, the good results reported have allowed its use to be expanded in many molecular biology laboratories around the world. The basic principle of this technique has been adapted for different applications such as specific methylation MLPA for the study of diseases related to epigenetic imprinting and digital MLPA that is coupled to next-generation sequencing equipment to increase the number of regions analysed in a single trial. With 10 years of experience in the use of this technique, the National Laboratory for Neonatal and High Risk Screening performs this review in order to analyse the principles, variants of the technique, data analysis, some applications, and advantages and disadvantages of MLPA compared to other available technologies.

Resumo A amplificação de múltiplas sondas dependentes de ligação (MLPA) é uma ferramenta valiosa no estudo das alterações do número de cópias para diferentes patologias de origem genética. A existência de uma ampla gama de kits comerciais, processamento laboratorial fácil e rápido, alta sensibilidade e, em geral, os bons resultados relatados, permitiram que seu uso se encontre expandido em muitos laboratórios de biologia molecular ao redor do mundo. O princípio desta técnica foi adaptado para diferentes aplicações como a MLPA metilação específica para o estudo de doenças relacionadas com o imprinting epigenético e a MLPA digital que é acoplada a equipamentos de sequenciamento de nova geração para aumentar o número de regiões analisadas em um único ensaio. Com 10 anos de experiência no uso da técnica, o Laboratório Nacional de Triagem Neonatal e de Alto Risco realiza esta revisão visando a analisar os princípios, variantes da técnica, análise dos dados, algumas aplicações, vantagens e desvantagens da MLPA comparado com outras tecnologias disponíveis.

Identification of a rare copy number polymorphic gain at 3q12.2 with candidate genes for familial endometriosis

Abstract Endometriosis is a complex disease that affects 10-15% of women of reproductive age. Familial studies show that relatives of affected patients have a higher risk of developing the disease, implicating a genetic role for this disorder. Little is known about the impact of germline genomic copy number variant (CNV) polymorphisms on the heredity of the disease. In this study, we describe a rare CNV identified in two sisters with familial endometriosis, which contain genes that may increase the susceptibility and progression of this disease. We investigated the presence of CNVs from the endometrium and blood of the sisters with endometriosis and normal endometrium of five women as controls without the disease using array-CGH through the Agilent 2x400K platform. We excluded common CNVs that were present in the database of genomic variation. We identified, in both sisters, a rare CNV gain affecting 113kb at band 3q12.2 involving two candidate genes: ADGRG7 and TFG. The CNV gain was validated by qPCR. ADGRG7 is located at 3q12.2 and encodes a G protein-coupled receptor influencing the NF-kappaβ pathway. TFG participates in chromosomal translocations associated with hematologic tumor and soft tissue sarcomas, and is also involved in the NF-kappa B pathway. The CNV gain in this family provides a new candidate genetic marker for future familial endometriosis studies. Additional longitudinal studies of affected families must confirm any associations between this rare CNV gain and genes involved in the NF-kappaβ pathway in predisposition to endometriosis.

Varietal Effect of Different Nitrogen Levels on Yield Attributing Characters and Quality Parameters of Wheat (Triticum aestivum L.) Crop

The field experiment was conducted at Agronomy Research FaTArm, Narendra Deva University of Agriculture and Technology, Narendra Nagar (Kumarganj) Faizabad (U.P.), during Rabi season of 2014-2015. The yield components like number of spikes m-2, spike length (cm), number of grain spike-1, grain weight spike-1(g), grain yield (kg ha-1), straw yield (q ha-1) and nitrogen contain in grain (%), nitrogen contain in straw (%), nitrogen uptake in grain (kg ha-1), nitrogen uptake in straw, protein content in grain (%) were maximum under 160 kg N ha-1 and among the varieties over PBW-373, NW-1014 being at par with HD-2327. The main aim of trial conducted that to know the different varietal parameters of wheat crop.

Epidemiological Studies on Leaf Blights of Maize Caused by Exerohilum turcicum (Pass.) Leonard and Suggs and Bipolaris maydis (Nisik. and Miyake)

The Field experiments conducted at Maize Research Station,Vagarai, Tamil Nadu, India during kharif 2019, 2020 & 2021 to study the disease development in relation to weather parameters, viz., temperature, relative humidity, rainfall with the leaf blights of maize. Observations on the spore load and disease grade were taken from 33rd standard week to 44th standard weeks at weekly interval. Increased spore load of 13 to 48 Nos. / Microscopic observations for TLB and 3 to 5 Nos./microscopic observations with the temperature ranges between 22-35?C, Humidity 48-72% and 19-24° Dew will lead to the TLB & MLB disease incidence with the grade of 1to 4 was observed. Based on the observations, the first appearance of leaf blight diseases were observed between 35th and 45th days after sowing with the grade of 1 (34th meteorological standard week). The disease grade increased from 1 to 4 as the age of the crop increases. There was a sudden increase in disease intensity because of increased scattered showers and increased relative Humidity (44th meteorological standard week). Temperature has not much influenced the disease development, since it was almost uniform throughout the cropping season in these three years. From the dataset, we would like to highlight that 34th and 44th meteorological standard weeks are highly critical for leaf blight disease development. Hence, spraying with mancozeb or zineb @ 2-4 g/l or propiconazole 25% EC @ 1ml/l during 34th and 44th meteorological standard week is recommended to manage the disease during kharif seasons in Tamil Nadu.

Panicle Architecture in Enhancing Grain Yield in Rice (Oryza sativa L.)

The architecture of the rice panicle is a crucial focus in breeding for both high yield and quality. It stands as a significant agronomic trait that influences the number of grains per panicle, playing a direct role in contributing to the overall yield of rice grains. Unravelling the genetic basis of panicle architecture is crucial for improving the grain yield in rice. In this study, the panicle architecture traits were meticulously dissected into five distinct components viz., number of primary rachis, number of secondary rachis, grain number on primary rachis, grain number on secondary rachis and total grain number. These components were systematically phenotyped in F2 and F3 population derived from a cross between DRR Dhan 48 and Maudamani. ‘DRR Dhan 48’ is a biofortified elite fine grain medium slender grain type cultivar with high zinc (22 ppm in polished rice) and low glycemic index (51.1). It has resistance to bacterial blight with the incorporation of xa5, xa13 and Xa21 in the background of Improved Samba Mahsuri. ‘Maudamani’ is a high yielding cultivar with high grain number and short bold grain type. Analysis of variance, histograms and boxplots revealed highly significant variation for the studied traits. Grain number in F2 population ranged from 29 to 333 grain with a mean of 162.72 whereas in F3, its varied from 49 to 368 with a mean of 184.17. Correlation analysis revealed significant correlation among the studied traits. The continuous variation observed in the population for panicle traits indicates that genetic control is governed by multiple minor loci. The presence of superior transgressive segregants highlights a complementary gene action influencing panicle architecture traits. The present investigation on panicle architecture offers scope for improving rice varieties and creating new germplasm resources and provides valuable information for further unravelling the genetic basis determining rice panicle architecture.

Analyzing the Variability in Maize (Zea mays L.) Genotypes Using Principal Component Analysis under Varied Ecosystems

Maize is a cereal cross-pollinated crop which belongs to the family poaceae. It is grown over a wider range of environment than any other cereal crop. The present study was conducted to study genetic variability using principal component analysis. Twenty maize genotypes were used in the study. Ten characters viz., plant height, ear height, days to 50 % tasselling, days to 50 % silking, ASI, cob length, number of rows per cob, hundred seed weight, shelling percentage and yield were recorded for the study. In the PCA, four principal components exhibited eigen value more than 1.0 exhibiting 84.54 percent of the variability for the characters under irrigated conditions. The PC1 negative loadings for the flowering traits, viz., days to 50 % silking (-0.3886), ASI (-0.3233) and days to 50 % tasselling (-0.2972) and positive loadings with higher contribution from cob length (0.4033), no of rows per cob (0.3480) followed by shelling percentage were observed. Positive loadings were exhibited by days to tasselling (0.4780), plant height (0.4670) and hundred seed weight (0.4518) in PC2. In PC3 positive loadings for the characters viz., number of rows per cob (0.5338), ASI (0.5226) and cob length (0.3299) and negative loadings for the traits grain yield (-0.3815), ear height (-0.2658) and plant height (-0.2437) and in PC4 positive loadings for plant height (0.4953), ear height (0.2546) and negative loadings were shown by days to grain yield (-0.6632), shelling percentage (-0.3751), ASI (-0.2369) and 50 % silking (-0.0648) were observed. In rainfed conditions, the two principal components had eigen value of more than 1. Under normal irrigated condition, PC1 and PC2 had eigen value of 5.689 and 1.6439. Plant height (0.3866), cob length (0.3726), number of rows per cob (0.3551), shelling % (0.3196), grain yield (0.2941), and hundred seed weight (0.2847) contributed positively to PC1. PC 2 had highest positive contributions from days to 50 % tasselling (0.5676), days to 50 % silking (0.5249), shelling % (0.3885), hundred seed weight (0.3763) and grain yield (0.2007). Negative loadings were shown by ear height (-0.1356) and plant height (-0.0093). Positive loadings were contributed by days to 50 % tasselling (0.5676), Days to 50 % silking, shelling percentage (0.3885) and hundred grain weight (0.3763).