Establishment and Modification of Ninety-seven Pneumococcal Serotyping Assays Based on Quantitative Real-time Polymerase Chain Reaction / 生物医学与环境科学（英文）

OBJECTIVE@#To establish and modify quantitative real-time polymerase chain reaction (qPCR)-based serotyping assays to distinguish 97 pneumococcal serotypes.@*METHODS@#A database of capsular polysaccharide ( cps) loci sequences was generated, covering 97 pneumococcal serotypes. Bioinformatics analyses were performed to identify the cps loci structure and target genes related to different pneumococcal serotypes with specific SNPs. A total of 27 novel qPCR serotyping assay primers and probes were established based on qPCR, while 27 recombinant plasmids containing serotype-specific DNA sequence fragments were constructed as reference target sequences to examine the specificity and sensitivity of the qPCR assay. A panel of pneumococcal reference strains was employed to evaluate the capability of pneumococcal serotyping.@*RESULTS@#A total of 97 pneumococcal serotyping assays based on qPCR were established and modified, which included 64 serotypes previously reported as well as an additional 33 serotypes. Twenty-seven novel qPCR serotyping target sequences were implemented in the pneumococcal qPCR serotyping system. A total of 97 pneumococcal serotypes, which included 52 individual serotypes and 45 serotypes belonging to 20 serogroups, could not be identified as individual serotypes. The sensitivity of qPCR assays based on 27 target sequences was 1-100 copies/µL. The specificity of the qPCR assays was 100%, which were tested by a panel of 90 serotypes of the pneumococcal reference strains.@*CONCLUSION@#A total of 27 novel qPCR assays were established and modified to analyze 97 pneumococcal serotypes.

Career Satisfaction among Community Pharmacists

Objective: This study's objective was to determine the level of career satisfaction among practicing Community Pharmacists (CPs) and the impact of various factors on career satisfaction. Methods: This cross-sectional study used a self-administered instrument to accessthe current level of career satisfaction among CPs. The survey instrument questions were adopted from previous studiesand werecontextualized to suit studyobjectivesand revalidated using face and content validation. The study instrument was distributed and collected. Results: A total of 172 responses from CPswere received, 112(65.1%) from males and 60(34.9%) from females. The difference in age categories was significantly profound in theless than 34 years old group being 113(65.7%) compared withthe more than 34 years old being 59(34.3%). Around 98(57%) were working in chain pharmacies and 74(43%) in independent pharmacy stores. The study result showed126 (73.3%) were unsatisfiedin their careers and 46(26.7%) were satisfied. Conclusion:Age and working experience are both major contributing factors to career satisfaction levels. Only 36.5% reported having higher career satisfaction levels among total participated CPs

Health-related Quality of Life among Community Pharmacists

Introduction:The role of community pharmacists (CPs) has kept on changing. Due to the evolving role of CPs, it is important to know its impact on the health-related quality of life (HRQoL) among CPs. Besides, the literature on the HRQoL of CPs is not being enriched, there is an urge to carry out a study to evaluate the HRQoL among CPs. This study aimed to examine the association between the socio-demographic factors and the HRQoLamongCPs. Methodology: A set of questions related to the HRQoL has been adopted from a pre-validated questionnaire, the Duke Health Profile and contextualized it to measure study outcomes. The target population in this study was the registered CPs that were practicing. The site of the study included all the chain and independent pharmacies. Statistical Package for Social Science (SPSS)version 22.0 was used to analyze the data and the significance level was set at p≤0.05. Variousdomains of the Duke Health Profile (DHP) were calculated using the provided formula Results: A total of 172 respondents were included in this study. Several HRQoL domains were significantly associated with socio-demographic factors. The univariate analysis illustrated that mental and general healthstatehad significant associations with age, practice setting was significantly associated with the mental health state, the salary was significantly associated with self-esteem and mental health was significantly associated with length of service and practice setting. Dysfunction dimensions of anxiety, anxiety-depression, and pain were significantly associated withgender. Conclusion: Gender, age, length of service, salary and practice setting were the risk factors for HRQoL among CPs

Predictors of Job and Workplace Satisfaction among Community Pharmacists

Aims: This study aimed to assess the current levels of job and workplace satisfactionamong community pharmacists (CPs) and to explore the factors that can affect their job and workplace satisfaction. Methods: A self-administered research tool (questionnaire) was developed based on previously published literature. After its reliability and validity measurements, the questionnaire was distributed to the target population and data was collected. Data wereentered into Statistical Package for Social Science (SPSS) ver. 22.0 and analyzed using descriptive statistics, chi-square,and multiple regression analysis. The level of significance (α) was set at 0.05.Results: CPs reported high satisfaction (76.7%) concerning their jobs. Only 23.3% of them were not satisfied with their current job. Univariate analysis showed that job and workplace satisfactionamong CPs was not significantly associated with gender, age, length of service, position and salary. However, the univariate analysis demonstrated a significant difference between practice settings and job and workplace satisfactionwith p= 0.013. Among the respondents, those who worked in the chain pharmacies (83.7%) expressed greater job and workplace satisfaction, which is 16% higher than those working in independent pharmacies (67.6%). Therefore, practice setting was shown as the predictor of job and workplace satisfactionamong CPs by using multiple regression analysis. Conclusion: Overall, CPswere relatively satisfied with their current job. Our results had reinforced previous studies that reported that practice settings can affect pharmacists’ job and workplace satisfaction.

A clinical study of new onset seizures in children aged 3-12 years

Background: Seizures are common in pediatric age group and less than 1/3rd of seizures in children are caused by epilepsy. Aim of present study was to analyze the etiological factors and clinical profile of new onset seizures in children aged 3-12 years.Methods: This was a prospective study done at a tertiary care center of south India. Authors analyzed 98 children aged 3-12 years presenting with new onset seizures. Proper history and investigations including EEG, and CT brain were done to confirm the etiology.Results: Out of 98 children 51 were 3-5 yrs of age and 47 were 6-12 years. 66.3% had generalized seizure and 33.6% had partial seizure. Partial seizure were more common in 5-12 years of age. Intracranial infections were the leading cause (73/98) which included neurotuberculosis (26/73), NCC (20), bacterial meningitis (12), cerebral malaria (8) and viral encephalitis (7). Intracranial infections' were the' leading cause in both age groups. Space occupying lesions were more common in 6-12 years (46.8%) as compared to 3-5 years (19.6%). NCC and Tuberculoma were the leading CT findings in both age groups. There was significantly high incidence of focal EEG changes in partial seizure group compared to generalized seizure group.Conclusions: Intracranial infections were leading cause of new onset seizures especially GTCS. Good clinical evaluation and judicious use of investigations should be ensured. CT brain and EEG are more likely to be informative in partial seizures than in generalized seizures.

Characterization of Actinobacillus pleuropneumoniae field strains antigenically related to the 3-6-8-15 group from diseased pigs in Japan and Argentina / Caracterización de cepas de Actinobacillus pleuropneumoniae antigénicamente relacionados al grupo 3-6-8-15 obtenidos de cerdos infectados naturalmente en Japón y Argentina

The objectives of this study were to determine the serovar of a collection of Actinobacillus pleuropneumoniae strains within the 3-6-8-15 cross-reacting group and to analyze their phenotypic and genetic properties. Based on the serological tests, forty-seven field strains of Actinobacillus pleuropneumoniae isolated from lungs with pleuropneumonia lesions in Japan and Argentina were found to be serovars belonging to the 3-6-8-15 cross-reacting group. By using a capsule loci-based PCR, twenty-nine (96.7%) and one (3.3%) from Japan were identified as serovars 15 and 8, respectively, whereas seventeen (100%) from Argentina were identified as serovar 8. The findings suggested that serovars 8 and 15 were prevalent within the 3-6-8-15 cross-reacting group, in Argentina and Japan, respectively. Phenotypic analyses revealed that the protein patterns observed on SDS-PAGE and the lipopolysaccharide antigen detected by immunoblotting of the reference and field strains of serovars 8 and 15 were similar to each other. Genetic (16S rDNA, apxIIA, apxIIIA, cps, cpx genes, apx and omlA patterns) analyses revealed that the apxIIA and apxIIIA genes of the field strains of serovars 8 and 15 were similar to those of the reference strains of serovars 3, 4, 6, 8 and 15. The results obtained in the present study may be useful for the development of more effective vaccines against disease caused by A. pleuropneumoniae by including the homologous antigens to the most prevalent serovars in specific geographical areas.

Los objetivos del presente estudio fueron determinar el serovar de una colección de cepas de Actinobacillus pleuropneumoniae pertenecientes al grupo 3, 6, 8, 15 de reacciones cruzadas y analizar sus propiedades fenotípicas y genéticas. En base a técnicas serológicas se determinó que cuarenta y siete cepas de A. pleuropneumoniae aisladas a partir de pulmones con lesiones de pleuroneumonía en Japón y Argentina pertenecen al grupo 3, 6, 8, 15. Mediante el uso de PCR basado en locus capsulares, veintinueve (96.7%) y una (3.3%) de los aislados japoneses fueron identificados como serovar 15 y 8 respectivamente, mientras que diecisiete (100%) de los aislados argentinos resultaron pertenecer al serotipo 8. Este hallazgo sugirió que los serovares 8 y 15 fueron los prevalentes dentro del grupo 3, 6, 8, 15 en Japón y Argentina, respectivamente. El análisis fenotípico reveló que los perfiles proteicos determinados por SDS-PAGE, y de antígenos lipopolisacáridos estudiados por inmunoblot, de las cepas de referencia y de campo de los serovares 8 y 15 fueron similares entre sí. El análisis genético (Í6S rDNA, apxIIA, apxIIA, cps, genes cpx, apx y los perfiles omlA) reveló que los genes apxIIA y apxIIIA de las cepas de campo de los serovares 8 y 15 fueron similares a sus homólogos de las cepas de referencia de los serovares 3, 4, 6, 8 y 15. Los resultados obtenidos en el presente estudio pueden ser útiles para el desarrollo de vacunas más efectivas contra la enfermedad causada por A. pleuropneumoniae, al posibilitar incluir antígenos homólogos a los serovares prevalentes en las áreas geográficas de interés.

Molecular identification and typing of 182 strains of Streptococcus pneumoniae isolated from children / 中国感染控制杂志

Objective To type Streptococcus pneumoniae(S.pneumoniae) isolated from children, and provide scientific basis for the correct selection of S.pneumoniae vaccine.Methods 182 strains of S.pneumoniae were collected from Children's Hospital of Hebei Province in 2014, species of strains were identified by polymerase chain reaction (PCR), types of strains were analyzed with multiplex PCR.Results PCR detection showed that cpsA gene amplification of 182 strains were all positive;multiplex PCR detection revealed that except 8 strains were not typed, the main types of the remaining 174 strains were 19 F (n=68, 37.36%), 19A(n=33, 18.13%), and 6A/6B (n=26,14.28%), the other types were 35B, 14, 6C/6D, 23F, 15B/15C, and so on.Conclusion The main types of 182 strains of S.pneumoniae are 19 F, 19A, and 6A/6B, which provide scientific basis for the correct selection of S.pneumoniae vaccine for this province.

Novel Pathogenic Variant (c.580C>T) in the CPS1 Gene in a Newborn With Carbamoyl Phosphate Synthetase 1 Deficiency Identified by Whole Exome Sequencing

Diagnosis of the urea cycle disorder (USD) carbamoyl-phosphate synthetase 1 (CPS1) deficiency (CPS1D) based on only the measurements of biochemical intermediary metabolites is not sufficient to properly exclude other UCDs with similar symptoms. We report the first Korean CPS1D patient using whole exome sequencing (WES). A four-day-old female neonate presented with respiratory failure due to severe metabolic encephalopathy with hyperammonemia (1,690 µmol/L; reference range, 11.2-48.2 µmol/L). Plasma amino acid analysis revealed markedly elevated levels of alanine (2,923 µmol/L; reference range, 131-710 µmol/L) and glutamine (5,777 µmol/L; reference range, 376-709 µmol/L), whereas that of citrulline was decreased (2 µmol/L; reference range, 10-45 µmol/L). WES revealed compound heterozygous pathogenic variants in the CPS1 gene: one novel nonsense pathogenic variant of c.580C>T (p.Gln194*) and one known pathogenic frameshift pathogenic variant of c.1547delG (p.Gly516Alafs*5), which was previously reported in Japanese patients with CPS1D. We successfully applied WES to molecularly diagnose the first Korean patient with CPS1D in a clinical setting. This result supports the clinical applicability of WES for cost-effective molecular diagnosis of UCDs.

Protective effects of Cordyceps sinensis polysaccharide CPS-A on angiotensin Ⅱ-induced injury of liver L02 cells / 中国药科大学学报

This study aimed at the molecular mechanism of Cordyceps sinensis polysaccharide-A(CPS-A) on angiotensin (Ang Ⅱ)-induced injury of L02 cells.The effect of AngⅡ and CPS-A on the proliferation of L02 cells was analyzed by MTT assay.PCR,Real-Time PCR and Western blot were also employed to determine the expression of IL-1β,AT1R,AT2R,NF-κB p65,TNFα and other inflammatory factors at mRNA and protein levels.The results showed that Ang Ⅱ and CPS-A could inhibit the proliferation of L02 cells by 1 × 10-5 mol/L and 200 μg/ mL,respectively.PCR,Real-Time PCR and Western blot showed that CPS-A could significantly down-regulate IL-1 β,TNF-α,NF-κB and AT1R.CPS-A has a good protective effect on AngⅡ-induced L02 cell injury.

Neonatal-onset carbamoyl phosphate synthetaseⅠdeficiency:a case report and literature review / 临床儿科杂志

Objective To explore the clinical characteristics of neonatal-onset carbamoyl phosphate synthetase I deficiency (CPS1D). Methods Clinical data and result of genetic detection of one neonate with CPS1D were retrospectively analyzed. The pertinent literature was reviewed. Results A 3-day old girl, with onset symptoms of nonspecific performance, such as poor feeding, less activity, tachypnea, and seizures. After fasting, anti-infection, and respiratory support etc. the condition was improved. However, the condition deteriorated and developed rapidly after feeding restarted. MRI showed extensive cerebral white matter lesions. Blood ammonia?>?500 μmol/L. Gene detection found two heterozygous mutations in pathogenic gene CPS1 in twentieth exon of c.2407C?>?G (p.803, R, G) and fourth exon C.323G?>?A (p.108, G, E), according to which CPS1D was diagnosed finally. Conclusions For neonate with normal birth, had feeding difficulty, seizures, and consciousness disorder after establishment of normal feeding, if blood ammonia level significantly increased, the blood and urine amino acids analysis and gene detection should be performed to confirm the diagnosis.

Impact of CPS1 Gene rs7422339 Polymorphism in Argentine Patients With Hyperhomocysteinemia

Abstract Carbamoyl phosphate synthetase 1 (CPS1) is a key gene in the first step of urea cycle and has been correlated with nitric oxide level and vascular smooth muscle activity. A functional single-nucleotide polymorphism C/A at position 4217 in CPS1 (National Center for Biotechnology Information SNP database no. rs7422339, T1405N) was reported to be associated with high homocysteine (Hcy) plasma values. Although genetic variants of methylenetetrahydrofolate reductase (MTHFR) gene are known to influence Hcy concentration, other genetic determinants of Hcy remain largely unknown. The association between the CPS1 rs7422339 and the risk of hyperhomocysteinemia in Latin American populations is unknown. Here, we study this association in 100 patients having hyperhomocysteinemia without MTHFR c.677C>T polymorphism and 100 controls. CPS1 rs7422339 was studied using polymerase chain reaction and enzymatic restriction. Comparisons of the CPS1 rs7422339 genotype distributions revealed a significant difference between groups (P = 2.3 × 10−3). Patients carrying polymorphic allele showed almost 3 times higher risk (odds ratio [OR] = 2.47) of hyperhomocysteinemia than wild-type allele, suggesting that rs7422339 SNP is associated with high Hcy levels in the Argentine population.

Qualidade microbiológica de lanches e salgados / Microbiological quality of finger food and snacks

Although finger food is convenient, it may be easily contaminated from the stage of preparation to themoment of consumption. This study aimed at evaluating the microbiological quality of finger food and sandwiches sold in Botucatu – SP, Brazil, by following the standards established by the Brazilian Health Surveillance Agency, ANVISA. The analysis was conducted according to APHA. A hundred and twenty two samples of meat, chicken, shrimp, cheese, and vegetable finger food and sandwiches were tested fromAugust 2008 to March 2009. Seventeen (13.9 %) samples of meat, cheese, vegetables and chicken were indisagreement with the ANVISA standards — some of them in more than one parameter. High counting of thermotolerant coliforms and coagulase-positive staphylococci were found in ten (8.2 %) and eight(6.5 %) samples, respectively. Salmonella spp. was detected in two samples (1.6 %). No Bacillus cereus andsulfite-reducing clostridia were isolated. Although only 10 samples (8.2 %) showed pathogenic bacteriacontamination, these results are relevant, since they indicate that the population is generally exposedto risks of acquiring foodborne diseases. Thus, the sanitary authorities might implement actions forsupervising the quality of the food sold in Botucatu, and to strengthen the food sellers to improve thehygienic conditions and be aware of the risks of food contamination.

Neste trabalho foi avaliada a qualidade microbiológica de salgados e sanduíches comercializados emBotucatu-SP, seguindo-se a legislação em vigor. As análises foram realizadas de acordo com APHA. Foramanalisadas 122 amostras de salgados de carne, frango, camarão, queijo e vegetais, e sanduíches no períodode agosto/2008 a março/2009. Dezessete (13,9 %) amostras de carne, queijo, vegetais e frango, e em algumasem mais de um parâmetro, estavam em desacordo com a legislação em vigor. Foram detectadas elevadascontagens de coliformes termotolerantes e estafilococos coagulase-positiva, respectivamente, em dez (8,2 %)e oito (6,5 %) amostras. Salmonella spp. foi isolada em duas amostras (1,6 %). As contagens de Bacillus cereuse de Clostrídio Sulfito Redutor não ultrapassaram os padrões da legislação. A maioria dos salgados mostrouresultados dentro dos padrões estabelecidos pela legislação. A presença de bactérias patogênicas comoS. aureus e Salmonella spp. foi demonstrada em 10 amostras (8,2 %); e este resultado é relevante, pois indicaque a população está exposta a riscos de doenças veiculadas por alimentos. Torna-se necessário colocarem prática a vigilância dos alimentos comercializados em Botucatu, incentivar a melhoria de condiçõesde higiene pelos comerciantes, e ter ciência dos riscos e das implicações da contaminação microbiológicados alimentos.

Formulation and comparison of in vitro release profile of hydrophilic and hydrophobic polymer based Naproxen matrix tablets.

The study was aimed to investigate the effect of polymer on the release profile of Naproxen from different percentages of HPMC 5cps and Kollidon SR based matrix systems. Different amount of HPMC and Kollidon SR were used to develop matrix builder in the four proposed formulations (F1-F4) for the study of release rate retardant effect at 25% and 35% of total weight of tablet matrix respectively. The tablets were prepared by direct compression method. The granules and tablets were evaluated for their physical properties and they did not show any significant variations and were found to have good physical integrity. The dissolution study of those proposed formulations were carried out in the simulated intestinal medium (pH 7.4) for 8 hours using USP paddle method with 50 rpm at 37±0.5⁰C. HPMC is hydrophilic and Kollidon SR is hydrophobic in nature. Statistically significant difference were found among the drug release profile from different percent of polymer and the release mechanisms were explored and explained with zero order, Higuchi and Korsmeyer equations. The release of Naproxen from F-1 and F-2 very closely followed Korsmeyer release kinetics where F-3 and F-4 best fitted with Higuchi model. The cumulative percent release of Naproxen was highest in F-2 containing 35% of HPMC. On the basis of results, it was found that the profile of F-1 formulation was the best among the four formulations. Between these two polymers, HPMC showed better percentage of release and Kollidon SR showed better release retardant effect.

Uso de PCR e sequenciamento do rDNA 16S para identificação de bactérias do gênero staphylococcus isoladas de mastite bovina / Identification of staphylococcus strains isolated from bovine mastitis by PCR and 16S rDNA sequencing

O objetivo deste trabalho foi identificar espécies de Staphylococcus (n=100) isoladas de mastite em rebanhos bovinos do Estado de Minas Gerais. Para esta finalidade foram utilizadas reações de PCR empregando oligonucleotídeos iniciadores descritos anteriormente para amplificar genes específicos de S. aureus (femA), S. intermedius (rDNA 16S) e S. hyicus (rDNA 16S-23S) e o sequenciamento do rDNA 16S. De acordo com as reações de PCR, 83 isolados foram identificados como S. aureus, 13 isolados como S. intermedius, dois como S. hyicus e dois isolados não foram identificados. Foram submetidos ao sequenciamento do rDNA 16S seis isolados identificados como S. aureus e os 17 restantes. Os seis isolados identificados como S. aureus confirmaram essa identificação. Dos outros 17 isolados, 13 foram identificados como S. chromogenes e quatro como S. hyicus, com similaridade igual ou superior a 99%. Baseando-se nos resultados da reação de PCR do gene femA e do sequenciamento do rDNA 16S, foram identificados 83 S. aureus, 13 S. chromogenes e quatro S. hyicus. Neste estudo os oligonucleotídeos iniciadores empregados na reação de PCR para S. intermedius não foram específicos, pois amplificaram também S. chromogenes; e os empregados na reação de PCR para S. hyicus não foram sensíveis, pois falharam na identificação de dois isolados de S. hyicus. A identificação definitiva das duas últimas espécies somente foi possível pelo sequenciamento do rDNA 16S.

The objective of this study was to identify the species of 100 isolates of Staphylococcus from mastitis in dairy cows from herds located in the state of Minas Gerais, Brazil. PCR reactions were carried out using specific primers described previously for S. aureus (femA gene), S. intermedius (16S rDNA) and S. hyicus (16S-23S rDNA spacer region). In addition, products of amplification of variable regions of the 16S rDNA gene of the strains were sequenced. According to the results of the PCR, 83 strains were identified as S. aureus, 13 as S. intermedius, two as S. hyicus and two isolates were not identified. The sequencing of 16S rDNA was applied to 23 strains identified by PCR amplifications: six S. aureus and the strains identified as S. intermedius (n=13), S. hyicus (n=2) or not identified (n=2). The sequencing of 16S rDNA confirmed the six strains as S. aureus. The others 17 strains were identified as S. chromogenes (13 isolates) and S. hyicus (four isolates). Each sample was related to a specie according to the smallest E-value and highest similarity (≥ 99%). The identification of S. hyicus and S. chromogenes was accomplished only by 16S rDNA sequencing.

Sequence analyses of the Cps2J gene in 9 strains of Streptococcus suis serotype2 isolated in Zhejiang province / 中国人兽共患病学报

To explore the molecular characteristics of Streptococcus suis serotype 2(ss2) isolated in Zhejiang province by deciding the variation loci and its variation frequency of Cps2J gene.The total DNA of 9 strains of ss2 isolated in Zhejiang province were extracted and amplifed by PCR. Then,the Cps2J fragments were cloned into plasmid carrier and completely sequenced after purification.Finally,the sequence results of all 9 ss2 isolates were compared with those obtained by other studies around the world.It was found that the open reading fragments of Cps2J in 9 SS2 isolates encoding 333 amino acids were 999 bp in length.Comparisons of this region among ss2 isolates revealed a similarity of between 98.8% and 99.9%, while the homology to ss1 strains varied between 56.8% and 57.0%.Our study shows the sequences of complete Cps2J segment are fairly stable and all these 9 ss2 strains of different sources possibly have the same evolutionary origin.

Promotion of Child's Rights and the Role of the Center for Prevention of Child Abuse in Korea

Child protective services, or the child protection system (CPS), forms the most front line safety net for abused and neglected children. The prerequisite for its activities lies in the awareness of promoting child's rights, which are well described in the United Nation's Convention on the Rights of the Child (UNCRC), and of the significant impact of child abuse and neglect as the origins to other social problems. Child protective services in Korea are carried on by 17 Centers for the Prevention of Child Abuse which have been opened since Octorber 2000 based on the amended Child Welfare Law which came into effect as of July 2000. The 24-hour telephone hotlines, numbered "1391", have been installed within the Centers. The counselors quickly respond to the reports, investigate the alleged cases of children and families with necessary services. This paper aims at the improvement of the understanding of child's rights and how the safety net, as the advocator for abused and neglected children, is functioning in Korea.

Immunogenicity of Vi capsular polysaccharide vaccine evaluated for three years in Korea

The immunogenicity of a single dose of Salmonella typhi(S.typhi) Vi capsular polysaccharide(CPS) vaccine was evaluated before, and at 1, 3, 12, and 36 months after vaccination. Eighty-five adults(20-28 years of age) and sixty-four children(8-16 years of age) received a single dose of 25 micrograms Vi CPS vaccine intramuscularly, and antibody titers to Vi CPS were measured by passive hemagglutination. Of 149 vaccinees, 138(92.6%) showed seroconversion at 1 month after vaccination, and then 138 out of 141(97.9%) did at 3 months. Of 137 vaccinees, 116(84.7%) maintained a persistent rise in Vi antibody titer 12 months after vaccination, and 55 out of 100(55.0%) had a 4-fold or greater rise at 36 months. No significant adverse reactions were observed. Booster injection may be needed 3-5 years after vaccination.

Uso masivo de la coproparasitoscopía con FAF / Masive use of coproparasitoscopy with FAF

El uso de solución preservadora y fijadora (FAF) de formas parásitas en la coproparasitoscopía de rutina para el diagnóstico de parasitosis intestinales, es considerado en este trabajo como la parte escencial para la realización de encuestas y trabajos de campo, cuando se desea tener una mayor confiabilidad, así como cuando se tienen que transportar lals muestras a un laboratorio de concentración. Su uso continuo ahorra material y equipo, y es adecuado para el procesamiento de los exámenes que más comunmente solicita el cliente

PREPARATION OF ~(35)S-LABEJJED CPS_1 cDNA PROBE FOR IN SITU HYBRIDIZATION / 解剖学报

The carbamyl phosphate synthetase I (CPS,) cDNA probe labelled with alpha ~(35)S-dATP was obtained by using modified nick translation technigue. The specific radioactivity of the obtained probe reached 0.9-1.5?10~8cpm/ug DNA. This probe could be satisfactorily used in RNA-DNA in situ hybrioization for detecting the gene expression of CPS_1 in liver tissue sections. The advantages of using ~(35)S instead of ~(32)P and ~3H for labelling DNA probe were discussed.

Extraction of Cuttlebone polysaccharides and purification of their active component CPS-1 / 第二军医大学学报

Objective:To extract the active polysaccharides from traditional Chinese herb-cuttlebone and to purify cuttlebone polysacchride salts 1(CPS-1),an active component of crude polysaccharides,so as to obtain refined natural active polysaccharides.Methods: Hot-water extraction method was optimized by orthogonal designing and was used to extract crude polysaccharides from cuttlebone.The total sugar contents of crude polysaccharides were determined.DEAE Sepharose F.F column and Sepharose CL-6B column were applied to separate CPS-1 from the crude polysaccharides.The active components were determined by animal experiments and Sephacryl S-300 column was used for further purification of CPS-1.HPLC was used to determine the purity of refined CPS-1 and the molecular weight of CPS-1 was determined by polysaccharides standard curve.Results: Crude polysaccharides were successfully extracted from cuttlebone.After purification with DEAE Sepharose F.F column,Sepharose CL-6B column and Sephacryl S-300 column and activity study,refined CPS-1 with average molecular weight(1?10~6) was obtained,and the sugar content reached 93.6%.Conclusion: Different extraction conditions have different extraction results for crude polysaccharides and different separation materials can separate the mixed polysaccharides by their molecular weights or charge characteristics.CPS-1 is a natural active polysaccharide extracted from cuttlebone.