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OBJECTIVE@#To provide prenatal diagnosis, pedigree analysis and genetic counseling for a pregnant woman who had given birth to a child featuring global developmental delay.@*METHODS@#A pregnant woman who underwent prenatal diagnosis at the Affiliated Hospital of Southwest Medical University in August 2021 was selected as the study subject. Peripheral blood samples were collected from the woman, her husband and child, in addition with amniotic fluid sample during mid-pregnancy. Genetic variants were detected by G-banded karyotyping analysis and copy number variation sequencing (CNV-seq). Pathogenicity of the variant was predicted based on the guidelines from the American College of Medical Genetics and Genomics (ACMG). Candidate variant was traced in the pedigree to assess the recurrence risk.@*RESULTS@#The karyotypes of the pregnant woman, her fetus, and affected child were 46,XX,ins(18)(p11.2q21q22), 46,X?,rec(18)dup(18)(q21q22)ins(18)(p11.2q21q22)mat and 46,XY,rec(18)del(18)(q21q22)ins(18)(p11.2q21q22)mat, respectively. Her husband was found to have a normal karyotype. CNV-seq has revealed a 19.73 Mb duplication at 18q21.2-q22.3 in the fetus and a 19.77 Mb deletion at 18q21.2-q22.3 in her child. The duplication and deletion fragments were identical to the insertional fragment in the pregnant woman. Based on the ACMG guidelines, the duplication and deletion fragments were both predicted to be pathogenic.@*CONCLUSION@#The intrachromosomal insertion of 18q21.2-q22.3 carried by the pregnant woman had probably given rise to the 18q21.2-q22.3 duplication and deletion in the two offspring. Above finding has provided a basis for genetic counseling for this pedigree.
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Criança , Feminino , Humanos , Gravidez , Masculino , Variações do Número de Cópias de DNA , População do Leste Asiático , Linhagem , Diagnóstico Pré-Natal/métodos , Cromossomos Humanos Par 18/genética , Feto , Mutação INDELRESUMO
The clinical data of a case of compound oxidative phosphorylation deficiency type 10 (COXPD10) caused by a new site mutation of MTO1 gene in the Department of Pediatrics, Affiliated Hospital of Southwest Medical University on December 29, 2020 were retrospectively analyzed.The patient was a 2 months and 19 days old boy of Han nationality.The main clinical manifestations were shortness of breath, hyperlactic acidemia, hyperammonemia and brain damage.Cardiac hypertrophy was not obvious.Heterozygous mutations at c. 344delA and c. 1055C>T sites in the MTO1 gene have not been reported in domestic and foreign literature.COXPD10 caused by MTO1 gene mutations may result in diversified clinical manifestations due to inconsistent mutation sites.For hyperlactic acidemia with unknown predisposing factors, early genetic examination should be conducted to confirm the possibility of COXPD10.
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Fine particulate matter (PM2.5) has attracted more and more attention in the field of environmental pollution and public health. Previous studies have found that PM2.5 can be inhaled and deposit in the airway and alveoli, and even spread to the whole-body tissues and organs through blood, resulting in various toxic effects. The malignant transformation of lung epithelial cells associated with long-term exposure to PM2.5 may play an important role in the occurrence and development of lung cancer. This paper reviewed recent studies on the mechanisms of malignant transformation of lung epithelial cells associated with PM2.5 exposure, and discussed the main biological mechanisms, including epigenetics, tumor microenvironment, and other biological pathways. Besides, the potential research directions of malignant transformation of lung epithelial cells associated with PM2.5 exposure were proposed. This work aims to provide a scientific basis and reference for public health management and air quality assessment.
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Objective:To establish the biology reference interval (RI) of peripheral blood procalcitonin (PCT) for children between 3 days and 6 years old in China.Methods:Totally 3 353 reference individuals with apparent health or no specific diseases were recruited in 18 hospitals throughout the country during October 2020 to May 2021. Reference individuals were divided into four groups: 3-28 days, 29 days - 1 year, 1-3 years and 4-6 years. Vein blood or capillary blood were collected by percutaneous puncture from every reference individual. The PCT level in serum and the capillary whole blood were assayed by Roche Cobas e601 and Norman NRM411-S7 immunoanalyzer. Outliers were deleted and 95th percentiles of every group were provided as RIs. Man-Whitney U test or Kruskal-Wallis test were used performed to assess the difference among different gender, age or method groups. Results:The difference of PCT distribution between male and female is not statistically significant, but the difference between serum and capillary whole blood is statistically significant. The differences between age groups are significant too. For Roche e601, serum PCT RI of 3-28 days group is <0.23 μg/L, 29 days - 6 years are <0.11 μg/L. For NRM411, Serum PCT RI of 3-28 days group is <0.21 μg/L, 29 days - 1 year: <0.09 μg/L, 1 - 6 years: <0.10 μg/L. For whole blood PCT, RI of 3-28 days group is <0.26 μg/L, 29 days - 6 years is <0.15 μg/L.Conclusions:Serum and capillary whole blood PCT have different RIs, however, capillary whole blood PCT testing is valuable in pediatric application. Children in 3-28 days show higher PCT levels than other age group. To establish the RIs and understand the differences among different groups are essential for the interpretation and clinical application of peripheral blood PCT testing results.
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Objective:To explore the clinical application of distal and proximal bilateral lateral pre-decompression in replant of digit degloving injury.Methods:From March 2012 to May 2021, 14 patients with 29 digits had replantation surgery of degloved soft tissue and severed digits in Section II, Department of Orthopaedics, Changzhou Medical District of 904th Hospital of PLA Joint Logistic Support Force. There were 2 types of injuries: degloving injury of soft tissue but with intact digital tip, and digital tip degloving injury with intact distal phalanx and nail bed. With the technique of distal and proximal bilateral lateral pre-decompression, pre-decompression incisions were made to the subdermal on both sides of the degloved skin over the proper palmar digital arteries. From where, the distal stump of the proper palmar digital arteries and nerves for anastomosis were found and had them anastomosed with the proximal proper palmar digital arteries and proper palmar digital nerves, then anastomosed digital pulp and digital dorsal veins (11 cases were direct anastomosis and 3 cases were bridged anastomosis). Follow-up was carried out by outpatient consultation, telephone and WeChat APP. The appearance of the digit body, the shape of digital pulp, the nails grow and the motions of the digits were observed, and the Evaluation Standard of Replantation of Severed Fingers by the Society of Hand surgery of Chinese Medical Association was used to evaluate the recovery of function.Results:All 29 segments of degloving digit survived. Small necrotic areas was found in 4 digits and healed after the change of dressing. The followed-up time was lasted for 3-36 months. There was no obvious atrophy found in all the digits. Appearances of digit pulp and nails were satisfactory with good sensational recovery for TPD at 6-10 mm, 7 mm in average. Motions of all the repaired digits were good. According to the Evaluation Standard of Replantation of Severed Fingers by the Society of Hand surgery of Chinese Medical Association, 23 digits were in excellent, 4 in good and 2 in poor, with a satisfactory rate at 93.1%.Conclusion:For a distal digit injury with intact distal soft tissue and relatively mild injury of blood vessels and nerves, using the method of distal and proximal bilateral lateral pre-decompression can help to achieve good appearance in survived digital pulp, good sensational recovery and good nail growth. An individualised treatment intra-and-after the surgery could offer an ideal therapeutic effect.
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Objective@#The purpose of this study is to investigate the status of freshmen s mental health literacy and its impact on mental health and to provide a reference for probing into the effectiveness and sustainality of curriculum education as the main channel of improvement of mental health literacy.@*Methods@#Mental health literacy questionnaire for Chinese adults and college students mental health screening scale were used to investigate 2 878 freshmen in a university in Wuhan. Mental health literacy was one of the indicators of course teaching evaluation. The curriculum intervention effect was investigated through pre-test, post-test and 10-month follow-up.@*Results@#The mental health literacy of freshmen (40.12±6.57) was higher than the national average level(35.81±8.06), but lower than the average level in terms of self-help mental health literacy(F=28.25，P<0.01). There were gender differences and urban-rural differences in mental health literacy. The mental health literacy of freshmen without psychological problems was significantly higher than that of freshmen with psychological problems(F=374.80，P<0.01). Structured course teaching significantly improved freshmen s mental health literacy, which were still significant after 10 months.@*Conclusion@#College freshmen s mental health literacy is generally great, and the mental health curriculum improved their mental health literacy.
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Lateral flow immunochromatography is a real-time detection technology based on chromatographic membranes and nano-markers. Because of its simplicity, fastness and low cost, it is widely used in many fields, such as biomedicine, pathogen detection and food safety. Traditional lateral-flow immunochromatography relies on the naked eye, that has lower sensitivity, and can only provide qualitative or semi-quantitative results. With the application of different types of markers and sensitive detection equipment, lateral flow immunochromatography has enabled the quantitative and multi-component detection of analytes. The purpose of this paper is to describe the latest research progress of lateral flow immunochromatographic detection systems in combination with nano-labels, chromatographic membranes and band detectors, with a view to providing a basis for the selection of markers, improvement of chromatography membrane materials and detection instruments.
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Objective The drug resistance and risk factors of methicillin resistant Staphylococcus aureus (MRSA) infected in the pediatric ward of the hospital were analyzed to provide a scientific basis for the con-trol of MRSA paediatric infection .Methods Totally 306 strains of Staphylococcus aureus were isolated from the pediatric wards in the Affiliated Hospital of Southwest Medical University from July 2015 to June 2016 , the bacterial species identification and drug susceptibility tests were performed by MicroScan WalkAway 96SI and the statistical analysis was carried out by WHONET 5 .4 softwate .The clinical data of 306 children were infected with staphylococcus aureus collected by retrospective survey ,the risk factors of MRSA infection were analyzed by the method of χ2 test and non-condition Logistic regression .Results A total of 90 strains of M R-SA were detected ,with a detection rate of 29 .41％ .MRSA was mainly isolated from sputum ,accounting for 74 .44％ ,followed by blood ,accounting for 10 .0％ .MRSA was mainly resistant to amikacin ,ampicillin ,eryth-romycin ,clindamycin and other antimicrobial agents (P< 0 .05) .Age< 1 years ,venous catheterization ,me-chanical ventilation ,tracheal intubation ,ventilator ,stay in ICU ,hospitalization time >7 d ,use of corticoste-roids ,low albumin ,and antibiotic use time >7 d were the main risk factors of MRSA in children (P<0 .05) . Admission to NICU ,venous catheterization ,mechanical ventilation and tracheal intubation were independent risk factors .Conclusion MRSA isolated from pediatric wards is resistant to many kinds of antibiotics .Infec-tion with MRSA is related to many factors .Hospitals should take corresponding measures to reduce the inci-dence of M RSA infection .
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Objective To investigate the effect of exogenous NPM mA to the subcellular localization of AKT and its significance in the proliferation in leukemia cells.Methods The co-immunoprecipitation assay was used to detect the interaction between NPM mA and AKT.The subcellular localization of NPM mA and AKT was observe by using the immunofluorescence experiment.The cells proliferation potential was assessed by CCK-8 assay.Results NPM mA could be combined with AKT in K562 cells.The immunofluorescence showed that NPM mA was mainly distributed in the cytoplasm.After co-tranfecting NPM mA and AKT into HEK293T cells,AKT was transformed from the dispersive distribution to cytoplasma.Additionally,the in vitro proliferation ability at 72 h in the K562 group was significantly enhanced compared with the K562 c1 and K562 mA groups(P＜0.01).After treating with AKT inhibitor(AKT inhibitor Ⅳ,AKT Ⅳ) for 48 h,the proliferation in the K562 c1 and K562 wt groups was remarkably inhibited,but which in the K562 mA group could still maintain growth(P＜0.01).Conclusion Exogenous NPM mA can change the subcellular localization of AKT,moreover regulates the in vitro malignant proliferation of leukemic cells.
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Objective To study the characteristics of lower respiratory tract infection by Streptococcus pneumonia in children aged from 0 to 5 years and drug resistance,so as to provide the rational basis for clinical treatment and epidemiology.Methods The medical data from 4 815 children aged 0 to 5 years with lower respiratory tract infection between September 2014 and September 2016 were retrospectively reviewed.Results Totally 343 strains of Streptococcus pneumonia were isolated from 4 815 sputum samples,with the isolation rate of 7.12％.Of the isolated streptococcus pneumonia strains,80.76％ were isolated from the children with pneumonia,9.33％ were isolated from the children with bronchitis,5.83％ were isolated from the children with capillary bronchitis,4.08％ were isolated from the children with bronchial asthma.There was no significant difference in the incidence of Streptococcus pneumonia infections between the genders of the children(P>0.05),but the incidence of Streptococcus pneumonia detection rates differed significantly in different seasons and children of different ages,the difference was statistically significant(P<0.05).The resistance rates of Streptococcus pneumonia to penicillin,erythromycin and clindamycin were 51.0％,97.9％ and 97.1％ respectively;The sensitivity rates to cefotaxime and vancomycin were 71.1％ and 100.0％ respectively.Conclusion The children aged less than 1 years old are at the high risk of Streptococcus pneumonia infection and highest detection rates in winter,it is necessary to perform the drug susceptibility testing in a timely manner so as to choice reasonable antimicrobial agents to control the condition.
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Objective To study the distribution and antimicrobial resistance of the bacterial strains isolated from lower respiratory tract in children,to provide evidence for better clinical management.Methods The medical data of children (0-5 years of age) with lower respiratory tract infection were retrospectively reviewed.The children were treated during the period from September 2014 to September 2016.Results Of the 4815 sputum samples collected from the lower respiratory tract of children,1582 (32.86％) had a positive bacterial culture.A total of 1614 strains of pathogens were identified.The most common bacterial pathogen was Haemophilus influenzae (9.66％),followed by Streptococcus pneumoniae (7.12％) and Moraxella catarrhalis (5.15％).The bacterial detection rate varied greatly with season and the age of children (P＜0.01).H.influenzae,M.catarrhalis and S.pneumoniae isolates showed lower resistance rate to cefotaxime,all ＜20.0％.Escherichia coli and Klebsiella pneumoniae isolates showed lower resistance rate to cefotetan,imipenem and piperacillin tazobactam,all ＜ 10％.Less than 20％ of the S.aureus strains were resistant to oxacillin.Conclusions H.influenzae is the most frequently isolated pathogen from lower respiratory tract in children aged 0 to 5 years in Neijiang Sichuan Province,followed by S.pneumoniae and M.catarrhalis.The detection rate of bacterial pathogens varies with season and the age of children.Antimicrobial agents should be selected rationally based on the results of antimicrobial susceptibility testing to reduce the occurrence of antibiotic resistance in clinical pathogens.
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<p><b>OBJECTIVE</b>To investigate the characteristics of gastrointestinal transit in chronic constipation patients depending on a small amount barium(SAB) gastrointestinal transit test.</p><p><b>METHODS</b>Imaging data of 1890 cases with chronic constipation diagnosed with Rome III( criteria undergoing the SAB gastrointestinal transit test at Department of Colorectal Surgery, The First Affiliated Hospital of Zhengzhou University from January 2009 to March 2015 were retrospectively analyzed. All the patients took 20 g medical barium sulfate diluted in porridge and erect position abdominal X-ray photographs were then taken at 4, 8, 12, 24 hours after meal and once in every 24 hour sequentially. Transit characteristics of whole gut and various segments, including stomach, small intestine, right hemicolon, left hemicolon and rectum were assessed according to the predetermined criteria.</p><p><b>RESULTS</b>All the 1890 patients showed good compliance with the complete test. Among these patients, 794 cases(42%) were diagnosed as normal transit and 1096 cases(58%) as gastrointestinal slow transit. Classified by the site of slow transit, 151 cases(8%) were abnormal gastric transit, 175(9%) cases were abnormal small intestinal transit, 986(52%) cases were slow right hemicolon transit, 1039 cases(55%) were slow left hemicolon transit, 139 cases(7%) were outlet obstruction. Among the 1096 patients with slow gut transit, 907 cases(83%) were multiple segments slow transit and 189 cases(17%) were single segment slow transit. Among 907 patients with multiple segmental slow transit, 668 cases(74%) were total colon slow transit, 61 cases(7%) were colonic slow transit combined with outlet obstruction, 138 cases(15%) were small intestine slow transit coincided with colon slow transit, 40 cases(4%) were total segments slow transit in the whole gut. Of 189 cases of single segment slow transit, 17 cases(9%) were unique gastric delayed empting, 37 cases(20%) were unitary small intestine transit dysfunction, 19 cases(10%) were right hemicolon transit defect, 78 cases(41%) were left hemicolon transit deterioration, 38 cases(20%) were outlet obstruction.</p><p><b>CONCLUSIONS</b>The characteristics of gastrointestinal transit test in patients with chronic constipation varied from each other. Majority presents multi-segment slow transit combined with colon slow transit. SAB gastrointestinal transit test is helpful for surveying the transit characteristics of each segment of gut and worth clinical generalization and application.</p>
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Humanos , Colo , Constipação Intestinal , Trato Gastrointestinal , Trânsito Gastrointestinal , Reto , Estudos Retrospectivos , EstômagoRESUMO
<p><b>OBJECTIVE</b>To explore the efficacy and safety of laparoscopic tunnel-like peri-anterior superior iliac spine suspension(L-TASISS) or combined with stapled transanal rectal resection (STARR) in the treatment of pelvic organ prolapse (POP) with outlet obstructive constipation (OOC).</p><p><b>METHODS</b>A total of 119 POP patients complicated with OOC( II( to IIII( stage evaluated by POP-Q) received L-TASISS or combined with STARR in the First Affiliated Hospital of Zhengzhou University from August 2010 to January 2015. Clinical and follow-up data of these patients were analyzed retrospectively and compared before and after operation.</p><p><b>RESULTS</b>Among 119 cases, 51 cases(42.9%) underwent L-TASISS alone, and 68 cases (57.1%) received L-TASISS combined with STARR. All the 119 patients were successfully operated without conversion to open surgery. The operation time was (67.8±10.9) minutes, the intra-operative blood loss was (10.3±3.8) ml, the indwelling catheter time was (3.6±1.1) days, and hospital stay was (5.1±1.8) days. One month after operation, abdominal wall pain or stress occurred in 15 cases, of whom 3 cases were improved by local block injection, 1 case by incision stitches release, the rest ameliorated spontaneously within 3 months after surgery. No potential ureterostenosis, hydroureterosis, internal iliac vascular thrombosis resulting from compression of the mesh and other complications related to the mesh were found. One year after operation, all the patients were followed up. The OOC remission rate was 78.2%(93/119), of whom 76 cases were satisfied and 17 patients were completely satisfied. One case(0.8%) with stress urinary incontinence did not improve. Fifteen cases(12.6%) with algopareunia or dyspareunia did not achieve remission, but there was no new algopareunia or dyspareunia case. Eleven patients (9.2%) presented recurrence of symptoms, of whom 9 cases(7.6%) complained of sensation of incomplete evacuation. Two cases(1.7%) were graded as POP-Q II(, and 1 case (0.8%) as POP-Q III( after surgery. Constipation Score of all the patients was 1.4±0.9 (compared to 7.8±3.6 preoperatively) according to Rome III( criteria. Enterocele occurred in 53 cases (44.5%) preoperatively corresponded with only 1 case (0.8%) after operation (χ(2)=64.77, P=0.000). One hundred and six cases (86.6%) with defecation difficulties and 87 cases (73.1%) with sensation of incomplete evacuation before operation were significantly improved after surgery, corresponding with 7(5.9%) and 9 (7.6%) symptomatic cases respectively (χ(2)=155.78, P=0.000). Three cases (2.5%) with preoperative fecal incontinence symptoms were improved after operation as well.</p><p><b>CONCLUSION</b>The procedure of L-TASISS or combined with STARR for POP patients with OCC has good short-term efficacy, and is safe and feasible.</p>
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Humanos , Pessoa de Meia-Idade , Constipação Intestinal , Procedimentos Cirúrgicos do Sistema Digestório , Métodos , Incontinência Fecal , Laparoscopia , Duração da Cirurgia , Prolapso de Órgão Pélvico , Cirurgia Geral , Prolapso Retal , Estudos Retrospectivos , Resultado do TratamentoRESUMO
Objective To study the drug resistance of multiple‐drug‐resistant Acinetobacter baumannii(MDR‐Ab) and its rela‐tive carbapenemases genes ,in order to provide references for rational use of antibacterial agents .Methods A total of 98 strains of Acinetobacter baumannii(Ab) were identified by using the MicroScan WalkAway96 automated microbial identification susceptibility testing system ,and the resistance genes ,including OXA‐23 ,OXA‐24 ,IMP ,VIM ,TEM and SHV ,were detected by using the poly‐merase chain reaction .DNA sequences of positive amplification products of the resistance gene were analysed .Results The drug re‐sistance rates of 98 strains of MDR‐Ab to penicillin class and cephalosporin class both were 100 .0% ,to imipenem and meropenem were 55 .1% and 54 .1% respectively ,to gentamicin ,amikacin and tobramycin were 100 .0% ,100 .0% and 87 .8% respectively ,to ciprofloxacin ,levofloxacin and gatifloxacin were 89 .8% ,91 .8% and 77 .6% respectively ,to sulfamethoxazole and rifampicin were 91 .8% and 100 .0% respectively ,to polymyxin B and polymyxin were 14 .3% and 11 .2% respectively ,to tetracycline ,minocycline and tigecycline were 100 .0% ,6 .1% and 4 .1% respectively .The results of resistance genes detection in 98 strains of MDR‐Ab showed that 70 strains carried TEM and OXA‐23 gene ,53 strains carried VIM gene ,41 strains carried IMP gene ,while OXA‐24 and SHV genes were not detected .DNA sequence analysis showed that the homology of OXA‐23 ,TEM ,IMP and VIM genes were 98% ,98% ,99% and 99% .Conclusion The condition of antibacterial resistance of MDR‐Ab in this area is very serious ,and TEM and OXA‐23 are the main drug resistance genes .Carrying multiple resistance genes is an important cause of MDR‐Ab resistance . The treatment of patients with Ab infection should be based on the results of drug sensitivity test for rational use of antibacterial a‐gents .
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Objective To study the protein fingerprinting of carbapenems resistant and susceptible Acinetobacter baumannii (Ab) strains ,investigate the clinical value of affinity distance in carbapenems resistant strains .Methods A total of 22 carbapenems resistant Ab strains and 18 carbapenems susceptible Ab strains were collected ,and bacterial protein fingerprinting was detected by surface-enhanced laser desorption ionization time of flight mass spectrometry (SELDI-TOF-MS) ,differentially expressed proteins was analyzed by Biomarker Wizard 3 .1 .Cluster analysis of differential expressed proteins was conducted on SPSS 19 .0 .Results The protein expression pattern of carbapenems resistant and sensitive Ab strains had significant difference (P< 0 .05) .Cluster anal-ysis showed carbapenems resistance Ab was given priority to with type A ,followed by type B .Conclusion The results of cluster a-nalysis carbapenems resistance of Ab protein fingerprinting could determine the distance of affinity relationship of Ab .It could pro-vide a theoretical basis for the infection and clinical epidemiology of Ab .
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The TGF-βsignaling pathway plays a crucial role in regulating cell proliferation, differentiation, and apoptosis. This pathway exerts either tumor-suppressing or tumor-promoting effects, which are cell-or context-dependent, making it simultaneously advanta-geous and disadvantageous in the process of carcinogenesis and tumor progression. Long non-coding RNAs (lncRNAs) do not encode proteins, but they are involved in the regulation of various signaling pathways and biological functions. Moreover, lncRNAs can induce tumor angiogenesis, as well as affect tumor proliferation, invasion, and metastasis by acting as oncogenes or tumor-suppressor genes. Recent studies revealed that some lncRNAs may be induced and regulated by TGF-βto form a complicated crosslinked regulatory net-work. This review focuses on the crosstalk between the TGF-βsignaling pathway and TGF-β-induced lncRNAs.
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Objective To evaluate the diagnostic value of serum retinol binding protein (RBP) on the early stage of type 2 diabet‐ic nephropathy by receiver operating characteristic(ROC)curve .Methods According to urinary albumin(mAlb)/urinary creatinine (UCr) ratio ,155 patients with type 2 diabetes were divided into simple diabetic mellitus group ,early stage of diabetic nephropathy group ,and clinical stage of diabetic nephropathy group ,while healthy people were recruited randomly during the same period as con‐trol group .RBP test were performed by using immunoturbidimetry .The diagnostic value of RBP on in the early stage of type 2 dia‐betic nephropathy were evaluated by analyzing the ROC curve .Results The concentration of serum RBP in the early stage of dia‐betic nephropathy was significantly higher than that in the group of simple diabetic mellitus and control group (P<0 .05) ,the area under the ROC curve of RBP in serum was 0 .770 ,and the cutoff value was 40 .95 mmol/L ,while the sensitivity and the specificity were 81 .0% and 95 .2% .Conclusion RBP was a good marker in detecting early renal damage .
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Objective To establish a simple ,rapid,highly specific and sensitive molecular detection of carbapenem-resistance acinetobacter baumannii(CRAB)OXA-23 genes,and this method is used to detect the multiple drug-resistant acinetobacter bau-mannii in our hospital,and the purpose is to know the antibiotic resistance of CRAB OXA-23 genes .Methods The loop-mediated isothermal amplification(LAMP)was established for detection of the CRAB OXA-23 genes,and a set of specific primers were de-signed by special software,PrimerExplorer version 4.The LAMP assay was developed on using SYBR Green Ⅰ for fluorescent chromogenic reaction substances,improved through a series of optimization tests,and through macroscopic observation and electro-phoresis test comparison results.At the same time,the application of LAMP was used to test 41 multiple drug-resistant acineto-bacter baumanniis which were collected from December 2013 to March 2014 in our hospitalized patients.Results The ladder ban-ding was produced in CRAB OXA-23 genes strains by the LAMP detection through electrophoresis test,however,no ladder ban-ding was observed in the others .The color of the amplification product in genes strain CRAB OXA-23 changed from orange to green by adding 1 μL SYBR Green Ⅰ,however it was still orange in others.The sensitivity of the LAMP detection in pure cultrue was 5 cfu/μL of the CRAB OXA-23 genes cells.Application of LAMP was used to separate multiple drug-resistant acinetobacter baumanniis from hospitalized patients ,32 strains were tested in 41 strains,the positive rate was 78.04%.Conclusion Separation of the CRAB OXA-23 genes carry rate is higher in our hospital ,and they have very high resistance of commonly used antibacterial drugs.The LAMP method to test OXA-23 gene of CRAB was established in this research was simple ,fast,sensitive and specific. Therefore,it is especially suitable wider use at the grass-roots unit,and it is of great significance for selecting reasonable choice of antibiotics by clinical doctor.
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BACKGROUND:Owing to containing large amounts of growth factors, platelet-rich plasma has been widely used in bone regeneration, wound healing, and so on, while few studies have been reported on cartilage tissue engineering. OBJECTIVE:To explore the effects of platelet-rich plasma on the proliferation and differentiation of chondrocytes, and the feasibility of constructing tissue-engineered cartilage by the combination of chondrocytes and platelet-rich plasma. METHODS:We detected the concentrations of transforming growth factor-β, insulin-like growth factor-1, platelet-derived growth factor and epidermal growth factor BB in the whole blood, platelet-rich plasma, and activated platelet-rich plasma. The rabbit articular chondrocytes were cultured in 10%, 15%, 20%, 30%platelet-rich plasma for 7 days, then the cellproliferation was tested by cellcounting kit-8, and cartilage-related genes (col agen type II, Aggrecan, Sox-9) were determined using QT-PCR. The chondrocytes/platelet-rich plasma composite was implanted subcutaneously into the rabbits, and the samples were harvested after 6 weeks of transplantation for histological examination. RESULTS AND CONCLUSION:The levels of different growth factors in the platelet-rich plasma were higher than those in the whole blood (P<0.05), but lower than those in the activated platelet-rich plasma (P<0.05). Platelet-rich plasma at different concentrations promoted the proliferation of chondrocytes. When the concentration of platelet-rich plasma was no more than 20%, the proliferation of chondrocytes showed a concentration-dependent manner. The 20%platelet-rich plasma showed the best effects to promote the expression of col agen type II (P<0.05), while the 15%platelet-rich plasma could maximize the expression of Sox-9 and Aggrecan (P<0.05). After transplantation of chondrocytes/platelet-rich plasma composite, cartilage-like tissue formed with cartilage lacuna-like structures, and rich extracellular matrix was found, which indicates platelet-rich plasma can be used as an injective scaffold in cartilage tissue engineering.
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Objective To establish a novel real-time PCR method to detect HCV RNA using Selfreporting duplex mutation primers.Methods The recombinant vector pMD18-T-HCV 5′-NCR was used as the calibrator.The Self-reporting duplex mutation primers were designed according to the gene sequence.And then the PCR reaction system was optimized and evaluated.The specificity,sensitivity and reproducibility of real-time PCR were estimated,The serum specimens from 90 cases(30 cases of HCV,30 cases of other viral hepatitis and 30 healthy volunteers) were tested with this real-time PCR; Results were compared with those obtained using a commercial TaqMan kit.Results The assay was established.It showed linearity over a wide range from 20 - 109 IU/ml.Intra-experimental coefficients of variation(CVs) were 1.37% -4.59%,and inter-experimental CVs were 1.58% -4.81%,respectively.There was no significant difference of HCV genome number tested by the two methods(R2 = 0.95) in 30 hepatitis C patients; HCV DNA was not detected in any serum samples of 30 healthy volunteers by the two methods.The specificity was 100%(60/60).All the samples in patients with clinically confirmed HCV infections showed HCV RNA positive.There wass good correlation between the quantitaive results and results obtained using the commercial TaqMan kit.Conclusions It is demonstrated that real-time PCR is a reliable,accurate and feasible assay for HCV.The establishment of this assay provided alternative technology for clinical diagnosis or therapeutic drug monitoring in the field of HCV infection and epidemiologic survey.