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1.
Article in Chinese | WPRIM | ID: wpr-921686

ABSTRACT

To study the active chemical components and mechanism of Liangfu Dropping Pills in treatment of gastrointestinal diseases. The UHPLC-Q-TOF-MS method was employed to analyze the components of Liangfu Dropping Pills in plasma. The protein targets of the absorbed compounds were predicted in the TCMSP database and the SwissTargetPrediction database. The targets associated with gastrointestinal diseases were collected from OMIM, CTD, GeneCards, and DrugBank. The common target genes between components and diseases were screened out for the building of protein-protein interaction(PPI) network in the STRING database. Metascape was used to carry out gene ontology(GO) and Kyoto encyclopedia of genes and genomes(KEGG) pathway enrichment analysis. Cytoscape was employed to construct the PPI network diagram and absorbed component-target network diagram. The molecular docking between the components absorbed in blood and potential key targets was performed by AutoDock vina 4.2.6 to screen and verify the main active components and targets. Twelve chemcial components were identified in Liangfu Dropping Pills, in which four components were absorbed in blood, including galangin, rhamnocitrin, galangin 3-methyl ether, and α-cyperone. These components acted on 189 common targets which were mainly involved in the cell responses to nitrogen compounds, organic cyclic compounds, and hormones, and enriched in the PI3 K-Akt signaling pathway, Foxo signaling pathway, and IL-17 signaling pathway. Molecular docking results showed that the four components had strong affinity with core targets. The material basis of Liangfu Dropping Pills treating gastrointestinal diseases may be galangin, rhamnocitrin, galangin 3-methyl ether, and α-cyperone. This study provides a theoretical basis for further development and application of Liangfu Dripping Pills.


Subject(s)
Drugs, Chinese Herbal , Gastrointestinal Diseases , Humans , Molecular Docking Simulation , Signal Transduction
2.
Article in Chinese | WPRIM | ID: wpr-828036

ABSTRACT

This study aims to reveal the pharmacokinetics of Shuganning Injection in normal rats. In this experiment,ultra-high performance liquid chromatography-electrospray-tandem mass spectrometry( UPLC-ESI-MS/MS) was used to establish an analytical method for simultaneous determination of chlorogenic acid,gardenioside,oroxylin A and baicalin in rat plasma. Then,the non-compartmental model( NCA) in Phoenix WinN onL in 6. 4 software was used to fit pharmacokinetic parameters. The methodological validation showed that the linear relationship of the components in rat plasma samples were good( r>0. 995). The recovery rate and matrix effect of plasma samples with low,middle and high concentration were 79. 14%-101. 4%. The intra-day and inter-day precision,accuracy and stability meet the requirements of biological sample analysis. The half-life( t1/2) of chlorogenic acid,gardenioside,oroxylin A did not change significantly and the area under blood concentration-time curve( AUC0-t) is proportional to the dose,which suggested that three components showed a linear kinetic characteristics,but baicalin showed nonlinear kinetic characteristics. Moreover,the retention time of each component in rats was short. The established UPLC-MS/MS quantitative analysis method is rapid,sensitive and accurate,which can be used for the determination of chlorogenic acid,gardenioside,oroxylin A and baicalin in rat plasma and pharmacokinetic study of Shuganning Injection.


Subject(s)
Animals , Chlorogenic Acid , Chromatography, High Pressure Liquid , Chromatography, Liquid , Plasma , Rats , Rats, Sprague-Dawley , Reproducibility of Results , Tandem Mass Spectrometry
3.
Article in Chinese | WPRIM | ID: wpr-827995

ABSTRACT

This project is to study the metabolites of Laportea bulbifera extract in rat feces. After the SD rats were gavaged with the extract(136 g·kg~(-1), according to the crude drug dose), the metabolites in their feces were detected by UHPLC-Q-TOF-MS~E technique, and the obtained mass spectrometry data was combined with UNIFI software for prediction. The prototype components and metabolites in rat feces were identified with reference materials and related literature. A total of 43 metabolites were identified(including 8 prototype components and 35 metabolites). The metabolic pathways mainly include monocaffeoylquinic acid(hydrogenation reduction, ring-opening cracking, sulfation, hydroxylation, glucuronidation), quercetin(O-C2 bond ring-opening cleavage, C2-C3 double bond reduction, rutin carbonylation) and so on. The metabolites and metabolic process of L. bulbifera extract in rat feces were clarified, which provided a basis for the study of the active substances and its mechanism of action.


Subject(s)
Administration, Oral , Animals , Chromatography, High Pressure Liquid , Feces , Plant Extracts , Rats , Rats, Sprague-Dawley , Urticaceae
4.
Article in Chinese | WPRIM | ID: wpr-802142

ABSTRACT

Objective: To compare in vitro dissolution behaviors of active ingredients (gastrodin,parishin A,p-hydroxybenzyl alcohol,parishin B and parishin C) in Gastrodiae Rhizoma powder with different particle size.Method: In vitro dissolution of Gastrodiae Rhizoma powder in different dissolution media (water,artificial gastric juice and artificial intestinal juice) were detected by stirring paddle method.Dissolution of these five components in Gastrodiae Rhizoma powder with different particle size was determined by HPLC,mobile phase was acetonitrile-0.1% phosphoric acid solution for gradient elution,column temperature was set at 40℃ and detection wavelength was 220 nm.Result: In water and artificial intestinal juice,the dissolution rates of five active components in three kinds of Gastrodiae Rhizoma ultrafine powders were higher than that of the fine powder and the finest powder;in artificial gastric juice,the dissolution rates of gastrodin and p-hydroxybenzyl alcohol in Gastrodiae Rhizoma ultrafine powders were higher than that of the other powders,and the dissolution rate of parishin A in Gastrodiae Rhizoma ultrafine powders was lower than that of the other powders.Conclusion: An appropriate degree of superfine grinding can promote the dissolution of active ingredients in Gastrodiae Rhizoma,but not as fine as possible.The dissolution medium has an obvious influence on the dissolution behaviors of active components,which provides a reference for screening optical particle size of Gastrodiae Rhizoma powder in clinical application.

5.
Article in Chinese | WPRIM | ID: wpr-802101

ABSTRACT

Objective: To study the protective effect of Gastrodiae Rhizoma ultrafine powder with different particle sizes on focal cerebral ischemia-reperfusion injury in rats, in order to provide theoretical basis for the application of ultrafine powder technology in related products. Method: The SD rats were randomly divided into six groups, namely sham-operated group, ischemia-reperfusion group, Gastrodiae Rhizoma ultramicro-powder Ⅰ group(1.2 g·kg-1), Gastrodiae Rhizoma ultramicro-powder Ⅱ group(1.2 g·kg-1), Gastrodiae Rhizoma ultramicro-powder Ⅲ group(1.2 g·kg-1), and nimodipine positive drug group(12.6 g·kg-1). The cerebral ischemia-reperfusion model was established after 7 days of drug administration. The focal ischemia reperfusion injury model was prepared by middle cerebral artery occlusion (MCAO) for 1 h followed by reperfusion for 24 h. The infarct size of brain tissue was measured by triphenyltetrazolium Chloride(TTC) staining. The activities of superoxide dismutase (SOD), malondialdehyde (MDA) and nitricoxide synthase (NOS) in brain homogenate and serum were measured by colorimetry. The contents of glutamate(Glu), aspartic acid(Asp) and glycine(Gly) in hippocampus were determined by liquid chromatography-tandem mass spectrometry (LC/MSMS). Result:The model group showed obvious signs of neurobehavioral deficit (PPPPPConclusion:Gastrodiae Rhizoma ultramicro-powder with different particle sizes has a protective effect on cerebral ischemia-reperfusion injury, and the Gastrodiae Rhizoma ultramicro-powder Ⅲ group has a more obvious protective effect than the the Gastrodiae Rhizoma ultramicro-powder Ⅰ group. The results showed that the protective effect of Gastrodiae Rhizoma ultramicro-powder on middle cerebral artery ischemia-reperfusion in rats was related to its particle size.

6.
Article in Chinese | WPRIM | ID: wpr-775130

ABSTRACT

OBJECTIVE@#To study the clinical characteristics, drug sensitivity of isolated strains, and risk factors of drug resistance in children with invasive pneumococcal disease (IPD).@*METHODS@#The clinical characteristics and drug sensitivity of the isolated strains of 246 hospitalized children with IPD in nine grade A tertiary children's hospitals from January 2016 to June 2018 were analyzed.@*RESULTS@#Of the 246 children with IPD, there were 122 males and 124 females. Their ages ranged from 1 day to 14 years, and among them, 68 (27.6%) patients were less than 1 year old, 54 (22.0%) patients were 1 to 2 years old, 97 (39.4%) patients were 2 to 5 years old, and 27 (11.0%) patients were 5 to 14 years old. Pneumonia with sepsis was the most common infection type (58.5%, 144/246), followed by bloodstream infection without focus (19.9%, 49/246) and meningitis (15.0%, 37/246). Forty-nine (19.9%) patients had underlying diseases, and 160 (65.0%) had various risk factors for drug resistance. The isolated Streptococcus pneumoniae strains were 100% sensitive to vancomycin, linezolid, moxifloxacin, and levofloxacin, 90% sensitive to ertapenem, ofloxacin, and ceftriaxone, but had a low sensitivity to erythromycin (4.2%), clindamycin (7.9%), and tetracycline (6.3%).@*CONCLUSIONS@#IPD is more common in children under 5 years old, especially in those under 2 years old. Some children with IPD have underlying diseases, and most of the patients have various risk factors for drug resistance. Pneumonia with sepsis is the most common infection type. The isolated Streptococcus pneumoniae strains are highly sensitive to vancomycin, linezolid, moxifloxacin, levofloxacin, ertapenem, and ceftriaxone in children with IPD.


Subject(s)
Anti-Bacterial Agents , Ceftriaxone , Child , Child, Preschool , Drug Resistance , Female , Humans , Infant , Male , Microbial Sensitivity Tests , Pneumococcal Infections , Streptococcus pneumoniae
7.
Article in Chinese | WPRIM | ID: wpr-771693

ABSTRACT

To investigate the absorptive characteristics of Inula cappa extract based on the rat everted intestinal sac method . Nine representative ingredients in I. cappa extract were selected as the study objects. An UPLC-MS/MS method was established to determine and detect their cumulative absorption amount for expounding the absorptive characteristics of ingredients in different intestinal sections. According to the results, the transport mechanism of 8 compounds showed passive diffusion by the reverted gut sac method. And scopolin was actively transported in the intestine. The best absorption site of chlorogenic acid was duodenum. The best absorption site of cryptochlorogenic acid, 1,3--dicaffeoylquinic acid, luteolin-7-glucoside and 3,4--dicaffeoylquinic acid were jejunum. The best absorption site of neochlorogenic acid, scopolin, 4,5--dicaffeoylquinic acid and 3,5--dicaffeoylquinic acid was ileum. The absorption of all the compounds was affected by pH and bile. All of the nine ingredients in I. cappa extract could be absorbed in intestines, but with differences in the absorption rate, the best absorptive site and mechanism, indicating that the intestinal absorption of I. cappa extract was selective.


Subject(s)
Animals , Chromatography, High Pressure Liquid , Intestinal Absorption , Intestines , Inula , Chemistry , Plant Extracts , Pharmacology , Rats , Rats, Sprague-Dawley , Tandem Mass Spectrometry
8.
Article in Chinese | WPRIM | ID: wpr-689883

ABSTRACT

The cerebral ischemia-reperfusion model was established to investigate the anti-oxidation and anti-apoptosis mechanism of Xinshao formula on the cerebral ischemia reperfusion injury in rats. SD rats were randomly divided into five groups: sham operation group, model group, and low, middle and high-dose Xinshao formula groups (0.31, 0.62, 1.25 g·kg⁻¹). After administration with Xinshao formula for 7 days, the rats were used to establish the cerebral ischemia-reperfusion model. The neurological behavior was evaluated. TTC staining was implemented to determine the volume of cerebral infarction. The levels of ROS, SOD, GSH-PX, NO and iNOS in serum were examined, and the mRNA and protein levels of Bcl-2, Bax and caspase 3 in hippocampal CA1 were detected by qRT-PCR, immunohistochemical assay and Western blot. It is found that Xinshao formula could significantly reduce the symptoms of nervous function and the volume of cerebral infarction in MACO rats. Compared with model group, the rats in Xinshao formula group showed increases in the activities of SOD and GSH-PX (<0.01), and decreases in the activities of iNOS and the contents of NO, ROS and MDA (<0.01). In addition, Xinshao formula could down-regulate the mRNA and protein levels of Bax and caspase 3 (<0.01), and up-regulated those of Bcl-2 (<0.01) in MACO rats. In conclusion, Xinshao formula showed aprotective effect on cerebral ischemia reperfusion injuryin rats, which may be associated with the promotion of anti-oxidation and anti-apoptosis.

9.
Article in Chinese | WPRIM | ID: wpr-335815

ABSTRACT

To investigate the metabolism of major components in Inula cappa by rat intestinal bacteria in vitro. I. cappa extract was incubated for 24 h with rat intestinal bacteria under anaerobic environment. After the samples were precipitated by n-butanol, UPLC-Q-TOF-MS/MS was applied for the qualitative analysis of the metabolites, combined with data software such as Metabolite Tools, Data Analysis and so on. The potential metabolites in rat intestinal bacteria were analyzed by comparing the total ion current of the test samples and blank samples and analyzing the quasi-molecular ion and fragment ion of all chromatograms. The results injected that fourteen metabolites were detected in rat intestinal flora. Various types of metabolic reactions happen to caffeoylquinic acid in intestinal flora, including isomerization, hydrolyzation, there were also methylation, hydrogenation and acetylation of caffeic acid. At the same time, a methylate of dicaffeoylquinic acid was also detected. Presumably, caffeoylquinic acids were gradually transformed into more hydrophobic metabolites with smaller molecular mass, which were better absorbed by the intestinal tract.

10.
Chinese Journal of Traumatology ; (6): 243-245, 2017.
Article in English | WPRIM | ID: wpr-330401

ABSTRACT

The Mycoplasma hominis infection is a rare postoperative complication after joint replacement. Based on our knowledge, there were only two cases reported by Korea all over the world currently. A case of postoperative Mycoplasma hominis infection after total knee replacement in our hospital was reported in this article. It was confirmed through mass spectrometer and Mycoplasma cultivation and treated by the first stage debridement, polyethylene insert replacement, and then drainage and irrigation combined with sensitive antibiotics after the operation. We observed that the C reactive protein (CRP) level correlates with the development of disease, while the erythrocyte sedimentation rate (ESR) remains at a high level, indicating the relevance between the Mycoplasma hominis infection caused by knee joint replacement and CRP. This study aims to report the case and review relevant literature.

11.
Article in English | WPRIM | ID: wpr-812565

ABSTRACT

The present study isolated 17 compounds from the tubers of Bletilla striata (Orchidaceae), using various chromatographic techniques. Their structures were identified based on their physical-chemical properties and spectroscopic analyses. Among them, two new 2-isobutylmalates, named bletimalates A (1) and B (2), together with other fifteen known compounds (3-17), were isolated and identified. Additionally, compounds 3, 4, and 8 were isolated from this plant for the first time.


Subject(s)
Malates , Chemistry , Molecular Structure , Orchidaceae , Chemistry , Plant Extracts , Chemistry , Plant Tubers , Chemistry
12.
Article in Chinese | WPRIM | ID: wpr-236058

ABSTRACT

To study the absorption characteristics of Xinshao extracts in Caco-2 cells. In this paper, human colon adenocarcinoma cell line Caco-2 cell model was established, and UPLC-MS method was applied to determinate the contents of five components of Xinshao extracts(albiflorin, gallic acid, caffeic acid, scutellarin and apigenin-7-O-glucronide) in cell lysates. This model was also used to study the effect of different drug concentrations, pH, time and temperature on the absorption of five components, investigate the transport of the five components of Xinshao extracts under the conditions with or without P-glycoprotein inhibitors, and predict the absorption mechanism of these five components in Caco-2 cells. The experimental results showed that the absorption of five components of Xinshao extracts in Caco-2 cells was time-dependent at 37 ℃, and concentration-dependent in the range of 0.5-12.5 g•L⁻¹, with a passive diffusion mechanism. At the pH of 4-7.4, the absorption of caffeic acid, scutellarin and apigenin-7-O-glucronide was significantly declined with the increase of pH(P<0.05). At the temperature of 4 to 37 ℃, the absorption of caffeic acid was declined with the increase of temperature, while the absorption of other four components was increased with the increase of temperature. Compared with the control group, caffeic acid and scutellarin cell absorption was significantly higher(P<0.05) after treatment with P-glycoprotein inhibitors(verapamil and cyclosporine A). The results indicated that, the absorption mechanism of five components in Xinshao extracts may be of passive diffusion, and the caffeic acid and scutellarin may be the substrates of P-glycoprotein.

13.
Chinese Journal of Microsurgery ; (6): 363-366, 2015.
Article in Chinese | WPRIM | ID: wpr-672234

ABSTRACT

Objective To discuss the efficacy of tantalum rod implantation combined with bone transplantation in the treatment of early stage osteonecrosis of the femoral head.Methods Twenty-four male patients were re cruited.Twelve patients were allocated to the tantalum rod and bone implantation group.The mean age was 35.7 years.Twelve patients were allocated to the tantalum rod implantation group.The mean age was 33.2 years.All patients with stage Ⅱ osteonecrosis.Results All patients were followed up with an average of 42.5 ± 6.7 months.The final evaluation was made according to the last follow-up data.In the tantalum rod implantation group,the average score of pre-operation and post-operation were 65.3±6.3,82.6 ± 5.3 respectively with a statistic difference (P < 0.05).Two of patients underwent total hip replacement before the end of this study,significant difference was observed in the aspect of radiographic progression (P < 0.05).In the tantalum rod implantation combined with bone marrow stem cell transplantation group,the average score was 92.2 ± 1.0 with a statistic difference (P < 0.05).There was no one of patients underwent total hip replacement before the end of this study.Significant difference was observed in the aspect of radiographic progression (P < 0.05).Between the two groups,a significant difference was observed in clin ical symptoms in favor of the tantalum rod implantation combined with bone transplantation group(P < 0.05).No significant difference was observed in the aspect of radiographic progression (P > 0.05).Significant difference was observed in the aspect of survivorship of the femoral head(P < 0.05).Conclusion Tantalum rod implantation combined with bone transplantation improves clinical symptoms and delay total hip arthoplasty than tantalum rod implantation.

14.
Article in Chinese | WPRIM | ID: wpr-330251

ABSTRACT

<p><b>OBJECTIVE</b>This study is to establish the fingerprint and find out the common chromatographic peaks of Inula cappa by HPLC.</p><p><b>METHOD</b>The HPLC analysis was performed on an Agilent Eclipse Plus C18 column (2.1 mm x 150 mm, 1.8 μm) with 0.1% fomic acid aqueous solution-0.1% fomic acid acetonitrile solution as mobile phase at a flow rate of 0.3 · mL(-1) · min(-1); The detective wavelength is 325 nm; The column temperature is 45 °C.</p><p><b>RESULT</b>The results indicated that 5 of 17 common peaks were identified . The similarity about 10 groups of Inulacappais is over 0.95.</p><p><b>CONCLUSION</b>This method is able to be a scientific basis of quality assessment according to its convenient and reliable.</p>


Subject(s)
Chromatography, High Pressure Liquid , Methods , Drugs, Chinese Herbal , Inula , Chemistry
15.
Article in Chinese | WPRIM | ID: wpr-299832

ABSTRACT

A UPLC method has been developed in the current investigation for simultaneous determination of nine chemical markers of Bletilla striata, 4-hydroxymethylphenyl beta-D-glucoside, blestroside, dactylorhin A, militarine, dihydrophenanthrene 5, gymnoside V, dihydrophenanthrene 1, benzylphenanthrene 3 and gymnosides IX. Separation was performed at 45 degrees C on an ACQUITY UPLC BEH C18 column (2.1 mm x 150 mm, 1.7 microm) with a gradient solvent system of acetonitrile-water as the mobile phase. The flow rate was 0.3 mL x min(-1), the detection wavelength was 280 nm. The results showed that the nine chemical markers could be well resolved and that in the selected linear range, all calibration curves of the nine chemical markers showed good linearity (r > or = 0.999 3). The recoveries (n = 6) were in the range of 98.15% - 102.2% and RSDs were between 2.1% - 3.6%. The data suggested that the developed UPLC-UV method had good reproducibility, robustness, and accuracy, which was suitable for the quality control of Bletilla striata. Applications of the method showed that the nine chemical markers had higher contents in the wild B. striata than in the cultivated ones.


Subject(s)
Chromatography, High Pressure Liquid , Methods , Drugs, Chinese Herbal , Magnoliopsida , Chemistry , Rhizome , Chemistry
16.
Article in Chinese | WPRIM | ID: wpr-299818

ABSTRACT

Using in vitro everted gut to investigate the intestinal absorption of the extracts from Polygonum orientale at different concentration. UPLC-MS/MS was used to detect the content of protocatechuic acid, isoorientin, orientin, vitexin, cynaroside, quercitrin, kaempferol-rhamnoside in different intestinal segments, then compared the results with the absorption of chemical components of extractive P. orientale in each intestinal segments, and calculated the absorption parameter. We took the statistic analysis with SPSS statistic software. The influence significance of each factors were analyzed to describe the character of absorption. The absorption of each component is linearity in different intestinal segments and different dose, and the square of coeficient correlation exceed 0.95, which consistent with zero order rate process. The K(a) increase along with the raised dosage of the extractive P. orientale (R2 > 0.95), indicated it is the passive absorption; different intestinal segments have different absorption. And the absorption trend in intestinal is duodenum, jejunum, ileum are greater than the colon. As ingredients are selectively absorbed in intestinal sac, the everted intestinal sac method is selected to assess the intestinal absorption charcteristics of ingredients of extractive P. orientale.


Subject(s)
Animals , Drugs, Chinese Herbal , Pharmacokinetics , Gastrointestinal Tract , Metabolism , Intestinal Absorption , Male , Polygonum , Chemistry , Metabolism , Rats , Rats, Sprague-Dawley
17.
Article in Chinese | WPRIM | ID: wpr-310993

ABSTRACT

This dissertation is to determine the biopotency of hemostat which processed in different places by establishing a bioassay method of Bletillae Rhizoma based on the thrombin time. Contrast test is the main methodology. Specifically, the reference substance of Bletillae Rhizoma is determined by comparing with the control substance of vitamin K1 using thrombin time, which is calibrated the Bletillae Rhizoma. The hemostatic biopotency is calculated by using the method of "parallel line assay method based on quantitative responses" (3.3) from different processed products. It indicates that there is a strong linear correlation between Bletillae Rhizoma and control drugs (Y = 66.332-23.913X, R2 = 0.995 3). The hemostatic biopotency of Bletillae Rhizoma from different processed products ranged between 821.93-1 187.53 U x g(-1) shown in the paper, and all of them can meet the requirements of the test. The methodology has an appropriate instrument precision (RSD 3.8%), intermediate precision (RSD 4.6%), repeatability (RSD 3.2%) and stability (RSD 3.7%). Therefore, it can be turned out that the methodology which established in the dissertation is good at determinating the hemostatic biopotency of Bletillae Rhizoma and it is reliable, simple and repeatable.


Subject(s)
Animals , Drugs, Chinese Herbal , Pharmacology , Reference Standards , Hemostatics , Pharmacology , Reference Standards , Orchidaceae , Chemistry , Rats , Rats, Sprague-Dawley , Rhizome , Chemistry , Thrombin Time
18.
Pakistan Journal of Pharmaceutical Sciences. 2014; 27 (3): 601-605
in English | IMEMR | ID: emr-142181

ABSTRACT

Tablets of oleanolic acid [OA] have been approved by SFDA in China as an adjuvant therapy for acute and chronic hepatitis. Co-occurring substances present in the tablets of OA and their hepatocytotoxicity have not yet been reported. In the current investigation, the crude OA drug was separated by repeated column chromatography. The structures of the isolated compounds were characterized by spectral analysis and the cytotoxicity of each compound was evaluated in vitro against the human normal liver cell L02 at concentrations from 0.125 to 1000 micromol/L using the MTT method. As a result, OA and its 11 co-occurring trace compounds including one new triterpenoid, 3-O- [4-oxopentanoyl]- olean-12- en-28-oic acid, were isolated and structurally characterized. Cytotoxicity tests indicated that these compounds were all non-toxic at concentrations up to 50micromol/L. Clear structure-activity relationship [SAR] was also observed. The results suggested that OA tablets of similar origin might not cause obvious cytotoxicity to the normal liver cell. The work may facilitate further SAR studies of OA-type triterpenoids.


Subject(s)
Liver/drug effects , Tablets , Triterpenes
19.
Chinese Journal of Epidemiology ; (12): 614-617, 2013.
Article in Chinese | WPRIM | ID: wpr-318339

ABSTRACT

<p><b>OBJECTIVE</b>To establish and evaluate a molecular diagnostic method for routine monitoring of four types of diarrheagenic Escherichia (E.) coli (DEC)and to study the distribution of four types of DEC isolated from diarrheal patients in Shanghai.</p><p><b>METHODS</b>DEC-PCR standard operation procedure(SOP)had been developed for DEC detection and isolation, using the Statens Serum Institute (SSI) DEC PCR kits with multiplex PCR technique after verification tests on reference strains. Diarrhea specimens from 3 clinical hospitals in Shanghai were tested from June to September, 2012.</p><p><b>RESULTS</b>Specificity of the PCR kit was 100% by verification on the 26 DEC reference strains. A total number of 218 DEC isolates, including 181 fermented lactose and 37 unfermented lactose were identified from the 1887 stool specimens of diarrhea patients, with positive rate as 11.6%. The most common pathogen(54.1%, 118/218)was enteropathogenic E. coli(EPEC), followed by enterotoxigenic E. coli(ETEC, 41.3%, 90/218), enteroinvasive E. coli (EIEC, 4.1%, 9/ 218) and Shigatoxin-producing E. coli(STEC, 0.5%, 1/218)in addition to 18 Shigella isolates. ETEC dominated in diarrhea patients with foreign residency, as well as 1/3 were perinatal stage of neonatal ETEC of all diarrhea cases under the age of 5, while EPEC dominated in the Chinese diarrhea patients especially among young kids under the age of 2.</p><p><b>CONCLUSION</b>Data was reliable after assessment on this molecular diagnostics and seperation procedures used for the routine monitoring on four types of DEC, while the diagnosis and reference ability of DEC regarding the laboratories net-working on food-borne pathogens need to be built up and improved.</p>


Subject(s)
Adolescent , Adult , China , Epidemiology , Diarrhea , Diagnosis , Epidemiology , Microbiology , Escherichia coli , Genetics , Escherichia coli Infections , Diagnosis , Epidemiology , Female , Humans , Infant , Male , Middle Aged , Pathology, Molecular , Sentinel Surveillance
20.
Chinese Journal of Pediatrics ; (12): 512-517, 2013.
Article in Chinese | WPRIM | ID: wpr-275664

ABSTRACT

<p><b>OBJECTIVE</b>To study the clinical and molecular characteristics of methicillin-resistant Staphylococcus aureus (MRSA) infection in children.</p><p><b>METHOD</b>A total of 37 MRSA strains were isolated from hospitalized patients in Children's Hospital of Fudan University from March 2009 to November 2011. The clinical characteristics were investigated by a cohort study. Furthermore, the mecA, Panton-Valentine leucocidin (PVL) genes were detected by polymerase chain reaction (PCR), and the genotypes of SCCmec were determined by multiplex PCR.</p><p><b>RESULT</b>(1) Among the 37 MRSA isolates, infections with 21 were acquired from hospital (HA-MRSA), and 16 isolates were acquired from community (CA-MRSA). (2) In the study, MRSA frequently caused respiratory tract infection, and most of the strains were isolated from intensive care unit (ICU). (3) CA-MRSA was most frequently associated with skin and soft tissue infections (SSTI), suppurative tonsillitis, even pneumonia and septicemia. HA-MRSA infection was more aggressive, most frequently associated with pneumonia, septicemia, and central nervous system (CNS) infections, such as meningitis. In children with fever caused by HA-MRSA or CA-MRSA infection, HA-MRSA showed a longer duration of fever, for 10.5 days. C-reactive protein (CRP) level caused by HA-MRSA (63.00 mg/L) was higher than CA-MRSA (9.50 mg/L) , and there were statistically significant differences between the groups (t = 2.5670, P < 0.05). However, there were no statistically significant differences between the groups in white blood cell count (WBC) or procalcitonin (PCT) level. (4) Among 37 MRSA isolates, the whole isolates were mecA gene positive (100%). SCCmec genotyping results showed that the most frequent SCCmec types were type III, 17 isolates, the others including type IV 8 isolates, type II1 isolates, nontypable 11 isolates, type I and type V were not found in this group. Therein, among 21 HA-MRSA isolates, SCCmec III was the most common, 15 isolates, type IV 1 isolates, nontypable 5 isolates; among 16 CA-MRSA isolates, SCCmec type IV was the most common, 7 isolates, type III 2 isolates, type II 1 isolate, nontypable 6 isolates. (5) Among the 37 MRSA isolates, 28 were PVL gene positive; and among 21 HA-MRSA isolates, 17 were PVL gene positive; Among 16 CA-MRSA isolates, 11 were PVL gene positive; There were no statistically significant differences between the groups (χ(2) = 0.735, P > 0.05) .</p><p><b>CONCLUSION</b>Compared with CA-MRSA, HA-MRSA infection was more aggressive, and induced higher C reactive protein; the dominant epidemic strains of CA-MRSA was SCCmec type IV, and HA-MRSA was SCCmec type III; the positive rate of PVL gene was high.</p>


Subject(s)
Adolescent , Anti-Bacterial Agents , Pharmacology , Bacterial Proteins , Genetics , Bacterial Toxins , Genetics , Bacterial Typing Techniques , Child , Child, Preschool , China , Epidemiology , Cohort Studies , Community-Acquired Infections , Epidemiology , Microbiology , Cross Infection , Epidemiology , Microbiology , DNA, Bacterial , Genetics , Female , Genotype , Humans , Infant , Infant, Newborn , Male , Methicillin , Pharmacology , Methicillin Resistance , Genetics , Methicillin-Resistant Staphylococcus aureus , Classification , Genetics , Penicillin-Binding Proteins , Staphylococcal Infections , Epidemiology , Microbiology
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