Analysis of the Results of MP-Ab-IgM Detection of Serum,Bronchial Lavage Fluid,Pleural Effusion and Cerebrospinal Fluid in Children Suspected of Mycoplasma Pneumoniae Infection / 现代检验医学杂志

Objective To study the infection of MP-Ab IgM (MP) in serum,bronchial lavage fluid,pleural effusion and cere brospinal from pediatric patients with Mycoplasma Pneumoniae Pneumonia (MPP).Methods From March to June 2013,278 cases of patients suspected MP infection in Children's Hospital Affiliated to Capital Medical University were selected and the MP-Ab IgM of thcire serum,bronchial lavage fluid,pleural effusion and ccrebrospinal fluid specimens were detected with the method of particle agglutination test.Results 181 cases (104 cases of boys,77 cases of girls) of different specimens in (MP) were positive.All the serum samples,278 cases were detected positive serum specimen of 170 cases,the overall detection rate was 61.2％ (170/278),144 cases of bronchial lavage fluid was investigated at the same time,check out the posi rive 62 cases,and the detection rate was 43.1％ (62/144).19 cases for pleural effusion samples at the same time,checked out the positive 13 cases,and detection rate was 68.4％ (13/19).15 cases of cerebrospinal fluid specimens was investigated at the same time,checked out the positive in 1 case,and the detection rate was 6.7％ (1/ 15).Positive rate between different age groups was respectively:6 months～1 year group 29.0％ (9/31),～3 year group 51.0％ (25/49),～5 year group 56.1％ (23/41),～16 year group 79.0％ (124/157),6 months～1 year witb other age groups was statistically significant difference (P＜0.05).In 0.5～1 year,～3 year,～5 year,16 year～four age groups,different types of MP-Ab IgM positive specimens of the same patients,the corresponding results for A:serum type (111 cases) 7,21,13,70.B:serum and bronchoalveolar lavage fluid type (45 cases) 0,2,5,38.C:Serum pleural effusion (7 cases) 0,1,2,4.D:serum and cerebrospinal fluid (1 cases) 0,0,0,1;E:serum and bronchoalveolar lavage fluid and pleural effusion type (6 cases):0,0,2,4.F:bronchoalveolar lavage fluid type (11) 2,1,1,7.Conclusion MP infection varied,have atrend of younger age,and has a tendency to increase year by year,and the patient's infection rates have different characteristics.Positive serum of different age groups was gradually increased with age,infants and young children positive detection rate increased significantlychildren with severe multiple in older children.

Microdeletion and Mutation of Y Chromosome in Full Sibling Identification / 法医学杂志

OBJECTIVES@#To explore the identification method of full sibling between two males with microdeletion and mutation of Y chromosome.@*METHODS@#DNA were extracted from two samples. The type testing of Y-STR and autosomal STR were performed. Full sibling between two individuals was calculated by IBS, ITO and discriminant functions methods.@*RESULTS@#There were 2 loci mutations existed in 33 Y-STR loci and one of the two samples had 19 loci deletions. The IBS of two samples was 53 and greater than the threshold which was 42; FSI was 1.36×10¹⁶ and far greater than 19. The discriminant function of full sibling-unrelated individual DFS2 was greater than DR2, which meant the two individuals tend to be full sibling.@*CONCLUSIONS@#The methods of IBS, ITO and discriminant functions of full sibling-unrelated individual can be used comprehensively to provide more reliable expert opinion in microdeletion and mutation of Y chromosome in full sibling identification.

Validation of GlobalFiler® PCR Amplification Kit and the STR Polymorphism / 法医学杂志

OBJECTIVE@#To test the technical parameters of GlobalFiler® PCR Amplification Kit for its application to forensic application value and to investigate the genetic polymorphisms.@*METHODS@#The validation was conducted in sensitivity, mixed samples, species specificity, adaptability, survivability, consistency, peak height balance and stability. The amplification and detection of the genomic DNA from 373 unrelated individuals from Beijing Han nationality were extracted by automation workstation.@*RESULTS@#Global-Filer® PCR Amplification Kit was adaptive to some mixed, degraded and inhibited samples. The power of sensitivity and adaptability and peak height balance showed well. The distributions of genotype frequencies for 21 STR loci in the population were all in accordance with Hardy-Weinberg equilibrium (P > 0.05). The PIC value of the 21 STR loci was among 0.536 to 0.940; the H value was among 0.558 to 0.933; the DP value was among 0.783 to 0.992; the PE value was among 0.243 to 0.874.@*CONCLUSION@#GlobalFiler® PCR Amplification Kit is suitable for criminal cases and DNA database in forensic practice. And 21 STR loci in Beijing Han nationality have high polymorphism, which have application value in forensic practice and population genetics.

Effects of cyclopamine on the proliferation and apoptosis of LNCaP cells and expression of the PCA3 gene in human prostate cancer / 中华男科学杂志

<p><b>OBJECTIVE</b>To explore the effects of cyclopamine on the proliferation and apoptosis of LNCaP cells and the expression of the PCA3 gene in human prostate cancer in vitro.</p><p><b>METHODS</b>LNCaP cells were treated with cyclopamine at the concentrations of 1, 5, 10 and 15 micromol/L for 24, 48 and 72 hours. The inhibitory effects of cyclopamine on the proliferation and apoptosis of the LNCaP cells were detected by MTT and flow cytometry respectively, the morphological changes of the cells observed by Hoechst 33258 staining, and the expression of the PCA3 gene determined by real-time fluorescence quantitative reverse transcriptase polymerase chain reaction (FQ-RT-PCR).</p><p><b>RESULTS</b>Compared with the blank control group, cyclopamine significantly inhibited the proliferation of the LNCaP cells at 5, 10 and 15 micromol/L (P <0.01), reaching IC50 at 10 micro mol/L at 48 hours. The apoptosis rates of the LNCaP cells at 24, 48 and 72 hours were 37.21%, 57.38% and 57.98% in the 10 micromol/L group and 21. 16% , 71.31% and 72.90% in the 15 micro.mol/L group, significantly different from those in the control (P <0. 01). The cell apoptosis showed a rising trend with the increase of cyclopamine concentration and acting-time, while the expression of the PCA3 gene was decreasing with the increased concentration of cyclopamine, significantly lower than that of the blank control group (P <0.01) , and extremely low in the 10 micromo/L group</p><p><b>CONCLUSION</b>Cyclopamine intervention at 10 and 15 micromol/L for 48 and 72 hours could significantly inhibit the at all time points. Proliferation and induce the apoptosis of LNCaP cells and reduce the expression level of PCA3.</p>

Modified TRIzol method for RNA and DNA co-extraction from blood / 法医学杂志

OBJECTIVE@#To establish a new method for RNA and DNA co-extraction from the same sample by TRIzol reagent.@*METHODS@#After the aqueous phase which contained total RNA was removed by traditional TRIzol method, the values of pH of the interphase phase and organic phase were adjusted. The DNA was precipitated with ethanol and purified with DNA IQ system. The purified DNA was measured in quality and quantity. As the template, it was amplified and typed by PCR-STR. The data was compared with that extracted by traditional TRIzol method.@*RESULTS@#The DNA extracted by this modified method showed a better result of quality and quantity than that by traditional TRIzol method and a good STR typing.@*CONCLUSION@#The modified TRIzol method is advisable and reliable to simultaneously extract both DNA and RNA from the same sample. It could be used for individual identification and paternity testing to satisfy the need of forensic science.

A new method of identifying the peripheral blood and the menstrual blood / 法医学杂志

OBJECTIVE@#To explore the tissue-specific gene expressions of the peripheral blood and the menstrual blood, and to search some specific factors to establish an effective method for identifying the peripheral blood and the menstrual blood.@*METHODS@#The specific products of the peripheral blood and the menstrual blood were detected by RT-PCR and separated by electrophoretic technology.@*RESULTS@#Beta-spectrin (SPTB) as one specific marker of peripheral blood and 18S rRNA as a kind of the housekeeping gene were expressed in both the peripheral blood and the menstrual blood. However, matrix metalloproteinase 7 (MMP7) as one specific marker of menstrual blood and human beta defensin 1 (HBD1) as one specific marker of vaginal discharge were only found in the menstrual blood.@*CONCLUSION@#There are differences of specific gene expressions between the peripheral blood and the menstrual blood. They could be accurately distinguished from each other by using the combination of fluorescence technology and RT-PCR to detect the specific identification of mRNA.

The study of the sensitive and specificity on HBV DNA replication by HBV-lP detecting / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To detect HBV-LP and HBV DNA of the patients with hepatitis B, and to study the sensitive and specificity of HBV-LP detecting and its evaluate on for clinical application.</p><p><b>METHODS</b>The ELISA was used for HBV-LP detecting and RT-PCR for HBV DNA detecting.</p><p><b>RESULTS</b>The sensitive and specificity of HBV-LP and HBV DNA were 64.89%, 99.68%, 60.63% and 100% respectively (P > 0.05); +LR, -LR were 202.78, 60.63, 0.3522 and 0.3937, and there were significance between +LR of the detecting (P < 0.005).</p><p><b>CONCLUSION</b>The sensitive and specificity of HBV-LP and HBV DNA detecting are considerable, +LR of HBV-LP are higher comparing HBV DNA. The HBV-LP is better serology index for detecting replication of HBV DNA.</p>

Human parvovirus B19 infection in patients with systemic lupus erythematosus / 中华风湿病学杂志

Objective To investigate the relationship between parvovirus B19 infection and systemic lupus erythematosus(SLE).Methods Sera from 51 patients with SLE and 20 normal controls were exam- ined for IgG and IgM antibodies against parvovirus BI9 by ELISA(produced by IBL Hamburg Company, Germany).Results Anti-Bl9 antibodies were found more frequently in sera from SLE patients than in normal controls,IgM and [gG antibodies were detected in 11(17.65%)and 9(21.57%)of 51 SLE pa- tients respectively.It was observed that the SLEDAI scores were higher in patients with B19 IgM positive group than those of the negative group(P＜0.05).The sera levels of ALT/AST were elevated more frequently in patients with BI9 IgM positive group than in negative group(P＜0.05).However,there was no association between the presence of anti-IgM and clinical manifestations in patients with SLE(P＞0.05).Five SLE pa- tients with positive B19 IgM were followed up for 2 years.SLEDA1 scores of these patients were markedly decreased(P＜0.05),but the levels of auto-antibodis(ANA,dsDNA)didn't change(P＞0.05).And the levels of ALT/AST decreased as usual.Conclusions It is suggested that parvovirus B19 infection may ex- acerbate the onset of SLE,but we eould not find a correlation between parvovirus B19 infection and prog- nosis of the disease.