OBJECTIVETo analyzed the relationship between lumbar endplate Modic area changes rate and low back pain by measuring MRI T2 sagittal image of lumbar endplate Modic area changes rate.

METHODSFrom December 2011 to June 2012,70 patients with low back pain in operation were evaluated on pain by VAS and function by JOA,and examined by MRI including 39 males and 31 females with an average age of (51.00 +/- 11.89) years ranging from 29 to 72 years old. Among them, 54 cases had lumbar endplate Modic changes involving 15 cases in types Modic I ,21 cases in type Modic II, 11 cases in type Modic III ,mixed type Modic in 7 cases (eliminated for too few cases). Modic area changes and corresponding vertebral area were measured on MRI T2 median sagittal. The areas of two ways were compared to yield the rate of changes for Modic, for multisegmental Modic changes to calculate the total ratios. A correlation was observed among JOA, VAS and the rate of Modic changes.

RESULTSThe correlation coefficient of change rate of Modic I with JOA score was r = -0.308, P = 0.048 < 0.05, there was a negative correlation;the correlation coefficient of change rate of Modic I with VAS scores was r = 0.428,P = 0.021 < 0.05, there was a positive correlation. The correlation coefficient of change rate of Modic II with JOA score was r = -0.375, P = 0.043 < 0.05, there was a negative correlation;the correlation coefficient of change rate of Modic II with VAS score was r = 0.352, P = 0.041 < 0.05, there was a positive correlation. The area change rate of Modic III had no significant correlation with low back pain degree (P > 0.05).

CONCLUSIONModic I and II area changes rate of of patients with low back pain is closely related to the degree of pain low back pain, Modic III area changes rate is not significant correlated to the degree of lower back pain.

Adult ; Aged ; Female ; Humans ; Low Back Pain ; diagnosis ; diagnostic imaging ; Lumbar Vertebrae ; diagnostic imaging ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Radiography

Objective To explore the genetic prenatal diagnosis method for acbendroplasia (ACH).Methods During May to November 2007, three ACH pedigrees were diagnosed at the Prenatal Diagnosis Center, Department of Obstetrics and Gynecology, Affiliated Drum Tower Hospital of Medical College, Nanjing University. In family 1, there was a 6-month-old male ACH infant. In family 2, the expectant mother, with 18 weeks of pregnancy, was an ACH patient. Amniocentesis was performed for prenatal diagnosis. The fetus of family 3 was diagnosed as ACH by ultrasound examination on the 39th week of gestation. Umbilical cord blood of this fetus was collected for examination. Totally, three methods, restriction enzyme (Sfc Ⅰ and Msp Ⅰ ) digestion analysis, denaturing high performance liquid chromatography (DHPLC) and sequencing analysis were performed simultaneously to detect the pathogenic mutation of flbroblastic growth factor receptor 3 (FGFR3) for the three ACH families. Results ( 1 ) The DHPLC detection: heteroduplex was detected in the patient of family 1 ; beth the patient and the fetus of family 2 showed heteroduplex results; the result of the fetus of family 3 was also heteroduplea. (2) The enzyme digestion analysis for the PCR products of 10 exon of FGFR3: after Sfc Ⅰ digestion, the PCR products of patients and the fetus of family 1 and 2 showed not only the band of 247 bp, but also bands of 162 bp and 85 bp. But their PCR products could not be digested by Msp Ⅰ , and it only showed the band of 247 bp. For the fetus of family 3, the PCR products could not be digested by Sfc Ⅰ , while after digestion by Msp Ⅰ , bands of 162 bp and 85 bp were shown up. The PCR products of the normal control could be digested by neither Sfc Ⅰ nor Msp Ⅰ. (3) The sequencing results: the heterozygote mutation of 1138 C→A was confirmed in the patient of family 1. The pregnant woman and her fetus in family 2 showed the same result. The heterozygote mutation of C→C was confirmed in the fetus of family 3. The site of 1138 was G homozygote in the normal control The three detection results of the fetus in family 2 were the same as that of the mother, which means that the fetus inherited the same pathogenic mutation from his or her mother. Conclusions Both DHPLC and restriction enzyme digestion analysis could detect the mutation of FGFR3 gene, but DHPLC is more rapid, convenient and sensitive. So DHPLC can be applied to genetic diagnosis and prenatal diagnosis for ACH patients.

Humans ; Leg Injuries ; surgery ; Male ; Middle Aged ; Surgical Procedures, Operative ; adverse effects ; Surgical Wound Infection ; etiology ; surgery

ObjectiveTo investigate the feasibitity of detecting S100A6 expression at mRNA level in endoscopic ultrasonography guided fine needle aspiration (EUS-FNA) pancreatic ductal adenocarcinoma (PDA) specimens and its diagnostic value in PDA.MethodsA total of 18 PDA specimens and 22 normal pancreatic specimens were collected. RNA was extracted for reverse transcription.The expression of S100A6 gene was examined by real-time polymerase chain reaction.The cut-off value of S100A6 expression at mRNA level in PDA diagnosis was established through receiver operating characteristic (ROC) analysis. 28 patients with pancreatic head masses were selected for EUS-FNA examination,and the value of S100A6 mRNA expression level in PDA diagnosis was prospectively evaluated. The expression of S100A6 protein in PDA tissue was determined by immunohistochemistry staining.ResultsS100A6 mRNA expression in EUS-FNA and surgical PDA specimens (0.05023±0.10120,0.02083 ± 0.02848) was significantly higher than that of normal pancreatic tissues (0.00164±0.00202),both P＜0.01.The expression of S100A6 in 22 EUS-FNA PDA specimens was significantly higher than that of 6 pancreatic benign disease biopsy specimens (0.00193 ± 0.00278,P =0.0009). There was no significant difference in S100A6 expression between 6 pancreatic benign disease biopsy specimens and normal surgical pancreatic samples (P=0.6143).When S100A6 mRNA expression in EUS-FNA specimens over 0.00525 was taken as positive diagnostic value,the sensitivity,specificity and accuracy in prospective pancreatic cancer diagnosis were 90.01％,100 ％ and 92.85 ％,respectively.ConclusionThe high expression of S100A6 mRNA in EUS-FNA specimens of PDA has good preoperative diagnostic value.

Background Our previous study determined that expressions of matrix metalloproteinases (MMPs) and tissue matrix metalloproteinase inhibitors (TIMPs)change in the conjuntival fibroblasts of conjunctivochalasis in vitro.To seek a suitable drug is very important in the prevention and treatment of conjunctivochalasis.Objective This study was to explore the effect of qijingmingmu soup on the expressions of MMPs and TIMPs in human conjunctival fibroblasts of conjunctivochalasis.Methods Twenty-four SD rats were randomized into two groups.Qijingmingmu soup was administration gastrically for consecutive 3 days,and normal saline solution was given in the same way in the control group.The blood was collected from aortaventralis and drug serum was prepared.Human conjunctival samples were obtained during the surgery of conjunctivochalasis relaxation and cultured in the DMEM containing 10％ fetal bovine serum,20％,15％,10％,5％ of drug serum and 8 ml/L epidermal growth factor(EGF) was added into the medium respectively.Enzyme-linked immunosorbent assay(ELISA)was used to detect the expressions of MMP1,MMP3,TIMP1 and TIMP3in conjunctival fibroblasts.Results Cultured cells grew well with the fusiform shape and showed the positive response for vimentin.The expression value of MMP1 (A value)in the cells was declined after administration of qijingmingmu soup.A significant difference was found in the expression of MMP1 among the control group,20％,15％,10％,5％ drug serum groups and EGF group(F=466.664,P＜0.05),and that in the 10％ ([9.92±0.14] mg/L) and 20％ ([11.87 ±0.11] mg/L) drug serum groups was significantly lowed in comparison with the control group([16.31±0.10] mg/L)(t=99.974,87.394,P＜0.05).The expression value of the MMP3in the cells in the various drug serum groups,EGF group and the control group was significantly different(F=158.168,P＜0.05),with a lower value in the 20％ drug serum group compared with the control group ([3.50±0.03] mg/L vs.[4.44 ± 0.11] mg/L) (t =21.991,P ＜ 0.05).Also,the significantly different expressing value of TIMP1 was seen among all the groups (F=183.508,P＜0.05),and expressing value of TIMP1 in the 15％ drug serum group was(1.88±0.06)mg/L,which was lower than(3.20±0.32) mg/L of the control group(t=10.353,P＜0.05).Furthermore,the expressing value of the TIMP3 in the cells was significantly different among the various groups(F=54.503,P＜0.05),and that of the 20％ drug serum group was (1.743±0.065)mg/L and it was significantly higher than (1.54 ± 0.05) mg/L of the control group (t =5.046,P =0.004).However,the expressing value of TIMP3of the 15％,10％ and 5％ drug serum groups was lower than that of the control group,respectively all at(P＜0.05).Conclusions Qijingmingmu soup drug serum at the concentration of 20％ can down-regulate the expressions of MMP1,MMP3,TIMP1 and up-regulate the expression of TIMP3 in human conjunctivochalasis bulbar conjunctival fibroblastsin vitro,which probably plays preventive and therapeutic effects on conjunctivochalasis.

Objective To discuss the detection rate of chromosomal abnormalities in women with different indications for invasive prenatal diagnosis(amniocentesis and eordocentesis), and the procedure-related complications. Metheds A retrospective analysis was conducted on 1264 women, who underwent invasive prenatal diagnosis (1082 amniocentesis and 182 eordocentesis), and the procedure-related complications were reviewed. Results The indications for invasive prenatal diagnosis in these 1264 women were: increased risk at prenatal screening (651, 51.5%), advanced maternal age (≥35) (318, 25.2%), abnormal foundings through uhrasonograph (136, 10.8%),history of adverse pregnancy (88, 6.9%), one or two abnormal serologic markers (52,4.1%), and chromosomal balance translocation carrier in either one of the couple(19, 1.5%). Thirty-seven cases were found to be chromosomal abnormalities with clinic significance and the indications for them were: ultrasonic abnormality (20/136, 14.7%); increased risk at prenatal screening (12/651, 1.8%); one or two abnormal serologic markers (1/52, 1.9%); history of adverse-pregnant (1/88, 1.1%)chromosomal balance translocation carrier in either one of the couple (3/19, 15.8%); advanced maternal age (0/318). Among the 1264 cases, 5 experienced spontaneous abortion and the procedure-related fetal loss rates were 0.28% for amniocentesis (3/1082) and 1.09% for cordocentesis (2/182), P=0. 154. The rate of complications after cordocentesis was significantly higher than amniocentesis (9.89 % vs 0.18 %, P= 0.0001). Conclusions Routine fetal karyotyping should be prompted after prenatal ultrasonographic abnormalities. However, invasive prenatal diagnosis due to advanced maternal age alone is controversial. Amniocentesis is the fist choice for invasive prenatal diagnosis.

OBJECTIVETo establish a rapid, sensitive and specific reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay for detecting human respiratory syncytial virus (RSV) in respiratory samples collected from children with acute respiratory infections.

METHODAccording to the conserved matrix gene sequences of respiratory syncytial virus subtypes A and B downloaded from GenBank, primers were designed and RT-LAMP assay was developed to detect RNA of RSV sensitivity of the RT-LAMP method was evaluated by using ten-fold serially diluted in vitro-transcribed matrix RNA fragments from RSV A and RSV B, respectively. Specificity of the RT-LAMP method was tested through cross-reaction with other RNA and DNA viruses. Then 5 RSV strains isolated from clinical specimens using tissue cultures were tested by RT-LAMP assay. A total of 101 nasopharyngeal aspirates from hospitalized patients with acute respiratory infections which had been tested by direct immunofluorescence assay (DFA), including 40 positive for RSV and 61 negative for RSV, were tested by RT-LAMP assay and by RT-nested PCR.

RESULTSensitivity analysis indicated that this RT-LAMP method was able to detect 1 copy/µl of RSV A and RSV B RNA, no amplification was shown in RT-LAMP with DNA or cDNA from other viruses in 60 min, revealed that the RT-LAMP assay is highly specific. Five RSV isolates confirmed as 4 RSV A and 1 RSV B previously were detected by RT-LAMP method as positive in 30 min. For those 101 specimens tested, 37 were RSV positive determined by RT-LAMP assay, as well as 35 RSV positive by RT-nested PCR. The total coincidence rate of RT-LAMP assay with DFA and RT-nested PCR in detecting RSV is 95.0%, 94.1% with Kappa value 0.895 and 0.871, respectively.

CONCLUSIONA new, sensitive, accurate and rapid method, RT-LAMP assay for detecting human respiratory syncytial viruses from nasopharyngeal aspirates was developed, which should be helpful in rapid detection of RSV from respiratory tract samples of children.

Acute Disease ; Child ; Child, Preschool ; DNA Primers ; Humans ; Infant ; Molecular Diagnostic Techniques ; Nasopharynx ; virology ; Nucleic Acid Amplification Techniques ; RNA, Viral ; isolation & purification ; Respiratory Syncytial Virus Infections ; diagnosis ; Respiratory Syncytial Virus, Human ; isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction ; Sensitivity and Specificity

Human metapneumovirus (hMPV) is associated with acute respiratory tract infections (ARTI) in all age groups. However, there is limited information of genetic analysis of hMPV circulating in Beijing. To learn the characteristics of structural protein genes of human metapneumovirus circulating in children in Beijing, sequence analysis of matrix (M), small hydrophobic (SH) and attachment (G) proteins of hMPV from 2006 to 2010 was performed. Phylogenetic analysis of nucleotide sequences of 42 full length M genes, 49 SH gene and 55 G gene revealed that the hMPVs from pediatric patients were divided into sub-genotypes A2, B1 and B. There were highly conserved identities among M gene, with 7 conserved mutations of amino acids between A and B genotypes which were fairly conserved in the same genotype A or B. The amino acid identities of SH were 60.7% to 64.4% between different genotypes, 93.3% - 100% among same sub-genotype and 84.7% - 88.7% between different sub-genotypes. Use of alternative transcription-termination codon, nucleotide deletion and insertion resulted in variable length of nucleotide and deduced amino acid of G protein. Amino acid identities within same genotype ranged from 81.5% - 100%, whereas sequence identities between two genotypes ranged from 34.0% - 38.6% at the amino acid level. A new cluster of G genes in sub-genotype B2 appeared due to the same mutations and insertion of two amino acids in G protein encoding genes amplified from specimens collected from 2008 to 2010. Prediction of antigen sites of SH and G protein indicated that the variation of antigen sites between different sub-genotypes existed.
Child ; China ; epidemiology ; Genetic Variation ; Genotype ; Humans ; Metapneumovirus ; genetics ; Paramyxoviridae Infections ; blood ; epidemiology ; virology ; Phylogeny ; Retroviridae Proteins, Oncogenic ; blood ; genetics ; Viral Envelope Proteins ; blood ; genetics ; Viral Matrix Proteins ; blood ; genetics

OBJECTIVEThe present study was designed to explore the practical application of the rapid etiological diagnosis by detecting specific IgM antibody against common respiratory viruses in children with acute lower respiratory infections (ALRI).

METHODClinical specimens including nasopharyngeal aspirates and serum of acute phase from hospitalized children were collected from 207 infants and children with acute lower respiratory infections from March 2009 to September 2010. Seven common respiratory virus antigens were identified from the collected nasopharyngeal aspirates by direct immunofluorescence assay (DFA). ELISA was used to detect specific IgM antibody against RSV, ADV, IFVA, IFVB and PIV, while indirect immunofluorescence assay (IFA) was used to detect specific IgM antibody against RSV, ADV, IFVA, IFVB, PIV1, PIV2 and PIV3 in collected acute phase serum.

RESULTThe overall positive rates to detect viral antigen by using DFA, ELISA and IFA was 67.6%, 57.5% and 39.6%, respectively. The consistent rate of ELISA and IFA versus accepted DFA were 21.7% and 31.4%, respectively. The average days from onset of the symptoms to blood sample collection for those with the consistent results by ELISA and DFA were 12.0 d for ADV, 9.6 d for PIV2, 9.5 d for IFV, and 5.3 d for RSV, respectively, and by IFA and DFA were 15.0 d for PIV3, 9.2 d for ADV, and 7.4 d for RSV, respectively. Among all age groups, the consistent rate of serum viral IgM and antigen detections was highest in children younger than 3 years old.

CONCLUSIONAlthough there were differences between serum IgM antibody and viral antigen detections, specific IgM antibody detection was of value in early and rapid etiological diagnosis of pediatric ALRI, especially for young children. It could provide serologic evidence of respiratory virus infection. The diagnostic rate of pathogen could be improved if it was used in combination with viral antigen diagnostic methods.

Antibodies, Viral ; analysis ; blood ; Antibody Specificity ; Antigens, Viral ; analysis ; Child ; Child, Preschool ; Enzyme-Linked Immunosorbent Assay ; Female ; Fluorescent Antibody Technique ; Humans ; Immunoglobulin M ; analysis ; blood ; Infant ; Male ; Nasopharynx ; virology ; RNA Viruses ; genetics ; isolation & purification ; Respiratory Syncytial Virus Infections ; diagnosis ; virology ; Respiratory Syncytial Viruses ; genetics ; isolation & purification ; Respiratory Tract Infections ; diagnosis ; immunology ; virology ; Sensitivity and Specificity

OBJECTIVESTo develop techniques which can be used to detect genetic material of measles virus from clinical samples to make diagnosis and differentiate atypical measles from other exanthematous infections early in clinical course.

METHODSA nested reverse transcriptase-polymerase chain reaction (RT-PCR) was developed to amplify gene fragment with the size of 301 bps from N gene of measles virus in throat swabs and urine samples collected from infants and children who were suspected measles cases. Before the test was used for clinical samples, preliminary tests were performed to determine the sensitivity and specificity of the test. The sensitivity of the test was determined by plaque assay using measles virus strain Edmonton and the specificity of the test was determined by cross-reaction with rubella virus, respiratory syncytial virus, influenza A and B viruses, enterovirus, adenovirus, human cytomegalovirus (hCMV), EB virus, and herpes simplex virus I. Serum specific IgM antibody against measles virus was also tested by ELISA.

RESULTSMeasles virus with the titer of 0.53 pfu could be detected by using the nested RT-PCR developed in this study. No amplification was found with the nested RT-PCR when rubella virus, respiratory syncytial virus, influenza A and B virus, enterovirus, adenovirus, hCMV, EB virus, and herpes simplex virus I were used as templates. Out of 116 throat swabs collected from suspected measles cases, 70 (60.3%) were measles RNA positive. For urine samples, 48 out of 74 (64.9%) were positive. Both throat swab and urine samples were collected simultaneously from 73 patients. Among those, 71 (97.3%) showed consistent results. Serum specimens were collected from 110 suspected patients. Among those, 65 (59.1%) and 61 (55.5%) were measles virus specific IgM antibody positive detected with ELISA kits from two different sources, respectively. Out of 110 sera samples, 106 (96.4%) showed consistent results. The consistency of the gene amplification and specific IgM antibody detection was 80.8% as shown by 84 out of 104 patients from whom throat swab and sera were collected at the same time.

CONCLUSIONThe data indicate that the nested RT-PCR developed in this study is sensitive and specific for detection of gene fragment of measles virus from clinical samples. The test is superior to the commonly used specific IgM antibody detection because of identifying gene material in early clinical stage, and even single clinical sample can be tested.

Humans ; Measles ; diagnosis ; genetics ; Measles virus ; genetics ; isolation & purification ; RNA, Viral ; genetics ; isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction ; Sensitivity and Specificity