Objective To evaluate the diagnostic value of autoantibodies to cell membrane associated with DNA (mDNA) in systemic lupus erythematosus (SLE) and the combination with other autoantibodies in the diagnosis of SLE. Method The anti-mDNA antibody had the characteristic pattern of perip-heral membrane fluorescence on cultured HL60. The same serum samples were detected for other antibo-dies of SLE. Pearson's Chi-square test was used for statistical analysis. Results This pattern was observed in 145 of 205 serum samples of SLE patients , but in 5 of 55 the serum samples of rheumatoid arthritis , in 10 of 45 primary Sjogren syndrome's patients and in 4 of 35 PM/DM and absent in 50 blood donors. The sensitivity and specificity of anti-mDNA antibody to SLE was 70.7% and 86.7%. The sensitivity and specificity of combined anti-mDNA antibody and ANA was 94.6% and 76.7%. The sensitivity and specificity of combined anti-mDNA antibody and anti-dsDNA antibody was 76.8% and 95.5%. The sensitivity and specificity of combined anti-mDNA antibody and anti-Sm antibody was 79.6% and 100%. The sensitivity and specificity of combined anti-mDNA antibody and AnuA was 93.0% and 100%. Conclusion This novel rapid immunofluorescence method can be a useful diagnostic test for SLE patients. Due to its high sensitivity and specificity, it is better than other diagnostic tests such as anti-dsDNA antibody and anti-Sm antibody for the diagnosis of SLE.

Animals ; Aorta ; pathology ; Apolipoproteins E ; deficiency ; Atherosclerosis ; metabolism ; pathology ; Female ; Glycosides ; isolation & purification ; pharmacology ; Intercellular Adhesion Molecule-1 ; metabolism ; Mice ; Plants, Medicinal ; chemistry ; Polygonum ; chemistry ; Vascular Cell Adhesion Molecule-1 ; metabolism

Objective:To establish an experimental model for intracellular antibacteria and endotoxin neutralization in vitro to detect the antibacterial and endotoxin neutralization activity of the muBPI_(25) protein.Methods: RAW264.7 cells were transfected with pcDNA3.1(+)muBPI_(36-259), and then were infected with intracellular bacterial of either G ~+/G~-to establish the experimental model of intracellalar antibacteria.The RAW264.7 cells were co-transfected with the pSecTag2B-muBPI_(36-259) and dual-luciferase reporter gene plasmids for establishment of the experimental model of endotoxin neutralization.Results:The experimental model of intracellular antibacteria confirmed that the muBPI_(25) protein could inhibit/kill Salmonella typhi.The experimental model of endotoxin neutralization indicated that the muBPI_(25) protein could neutralize endotoxin.Conelusion: We firstly demonstrate that murine BPI N-terminal functional fragment(muBPI_(25) protein)can inhibit/kill Salmonella typhi,and can neutralize, its lysating product, endotoxin.

OBJECTIVETo investigate the relationship between genetic polymorphism in exon 12 C1236T, exon 21 G2677T/A and exon 26 C3435T of the multidrug resistance 1 (MDR1) gene and the risk of childhood acute lymphocytic leukemia (ALL).

METHODA total of 176 patients with ALL and a cohort of 170 matched healthy subjects were included. SNaPshot SNP typing was used to determine the genotypes of MDR1 C1236T, G2677T/A, C3435T. Based on the clinical data, the relationship between genetic polymorphism of MDR1 and the risk of childhood ALL was analyzed.

RESULTThere was significant difference in the distribution of genotype of MDR1 C3435T between the group of controls and cases. The mutant homozygous TT genotype was found to be associated with occurrence of ALL (P = 0.000; OR = 4.504). The data show evidence of pairwise linkage disequilibrium between the three common SNPs (C1236T-G2677T/A-C3435T). The haplotypes of TTT, TGC, CGC and CAC were predominant. The haplotype CGT distributed significantly differently between the groups of controls and cases (P = 0.034). The frequency of the haplotype TTT/TTT in the high risk group was higher than the other groups (P = 0.037).

CONCLUSIONThe present findings suggest that 3435C→T polymorphism in MDR1 gene may be a genetic susceptibility factor for ALL. The haplotype of MDR1 (C1236T-G2677T/A-C3435T) could be the clinical parameter at diagnosis.

ATP Binding Cassette Transporter, Sub-Family B ; ATP-Binding Cassette, Sub-Family B, Member 1 ; genetics ; Acute Disease ; Asian Continental Ancestry Group ; genetics ; Child, Preschool ; China ; ethnology ; Exons ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Haplotypes ; Humans ; Infant ; Linkage Disequilibrium ; Male ; Polymorphism, Single Nucleotide ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; genetics ; Risk Factors

BACKGROUNDIn recent years, the incidence of unexplained sudden death has risen significantly across the world. However, it occurred suddenly, often in young apparently healthy individuals and almost 50% of the patients did not have any warning signals or symptoms. Therefore, the prodromal symptoms before the incident are extremely important for early prediction of sudden death. In this article, we aimed to explore the value of prodromal symptoms for unexplained sudden death and whether the prodromal symptoms have a predictive function to unexplained sudden death (USD) without underlying diseases.

METHODSA total of 208 sudden death cases were selected for the survey in the Emergency Department of Peking University Third Hospital from January 2006 to December 2009 and their medical records were reviewed. The patients were divided into two groups, 65 patients had underlying diseases while 143 had not underlying diseases. In the meantime, their prodromal symptoms were collected and compared, prodromal symptoms including chest distress, dyspnea, syncope, fever, headache, vomiting, etc.

RESULTSPatients with underlying diseases were compared to those without underlying diseases associated with sudden death; there was no significant difference in gender and age distribution. Among the 208 cases, 39 cases (18.75%) had prodromal symptoms, patients with underlying diseases had prodromal symptoms in 12 cases (18.46%), while patients without underlying diseases had prodromal symptoms in 27 cases (18.88%). The difference between the two groups with prodromal symptoms was not statistically significant (P > 0.05).

CONCLUSIONSProdromal symptoms are extremely important warning signals in the occurrence of USD. It has equally important predictive value for patients both with and without underlying diseases, especially in predicting sudden death caused by cardiopulmonary and neurological diseases.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Death, Sudden ; epidemiology ; etiology ; Female ; Humans ; Male ; Middle Aged ; Prodromal Symptoms

OBJECTIVETo study the morphologic features and clinical significance of atypical small acinar proliferation (ASAP) suspicious but not diagnostic of cancer in prostatic biopsies.

METHODSThe slides of 11 cases of prostatic needle biopsies collected during a two-year period with the diagnosis of ASAP were reviewed. Immunohistochemical study for 34betaE12, p63 and P504S was performed on the archival paraffin sections.

RESULTSAll the 11 ASAP cases were characterized by the presence of a few compacted small acini in the prostatic stroma. Six cases had acini of less than three in number. The acini were round or slightly irregular in shape. The nuclei were enlarged, round or irregular, arranged in single layer and focally separated by broad interval. The nucleoli were usually prominent. Cytoplasm was amphophilic or pale and the lumen border was often well-defined. Basophilic mucus was also seen in some of the lumen. Immunohistochemical study for 34betaE12 and p63 was negative, while that for P504S was positive. In 4 of the 11 cases, the acini were more than three in number, round or slightly irregular, but without cytologic atypia. The nuclei were slightly enlarged with small or inconspicuous nucleoli. Immunohistochemical study for 34betaE12 and p63 was negative or at most focally positive. P504S staining was either negative or weakly positive. Second repeat biopsy was carried out in all cases, and 4 of them (36%) showed definite adenocarcinomatous changes. The positive cases were those with fewer acini but definite cytologic atypia in the initial biopsy.

CONCLUSIONSASAP is a morphologic interpretation closely associated with prostatic adenocarcinoma. The histologic features are suspicious of but not diagnostic of cancer, due to insufficient criteria in terms of acinar number, cytologic or architectural abnormalities. The positive rate in subsequent repeat biopsy is higher than that for cases with usual nodular hyperplasia.

Adenocarcinoma ; enzymology ; pathology ; Aged ; Biopsy ; Diagnosis, Differential ; Humans ; Male ; Middle Aged ; Prostate ; enzymology ; pathology ; Prostatic Hyperplasia ; enzymology ; pathology ; Prostatic Intraepithelial Neoplasia ; enzymology ; pathology ; Prostatic Neoplasms ; enzymology ; pathology ; Racemases and Epimerases ; metabolism

The aim of this study was to separate and amplify CD4(+)CD25(+)Treg cells from splenocytes of sensitized mice. The percentage of CD4(+)CD25(+)Treg cells was detected by flow cytometry in sensitized and normal control mice. CD4(+)T, CD4(+)CD25(+)Treg and CD4(+)CD25(-) T cells were isolated from mouse splenocytes by MACS. CD4(+)CD25(+)Treg cells were expanded in vitro cultures in addition of CD3/CD28 MACSiBead and IL-2. The activity of cells was detected with 0.4 trypan blue staining. The purity of cells after sorting, the main surface marker and the level of Foxp3 were detected by flow cytometry. The results showed that CD4(+)CD25(+)Treg cell proportion was higher in sensitized mice than normal control mice (P < 0.05). The average purity of CD4(+)CD25(+)Treg cells was 87. The activity of these cells was more than 97, and the expression of Foxp3 in these cells was high. The amplification multiples achieved 42 times after 2 weeks in vitro. The percentage of CD4(+)CD25(+) regulatory T cells was 85.32, and the expression of Foxp3 decreased from (76.92 ± 1.72) to (75.33 ± 2.11) (P > 0.05). It is concluded that the sorting of CD4(+)CD25(+)Treg cells is isolated successfully by MACS without affecting the vitality of target cells. The amplification of CD4(+)CD25(+)Treg cells is successful in vitro. Expression of surface markers and Foxp3 gene does not obviously change after amplification, so that to establish a practical method to recover and enlarge the amount of CD4(+)CD25(+)Treg cells in good condition.
Animals ; CD4 Antigens ; biosynthesis ; Flow Cytometry ; Forkhead Transcription Factors ; metabolism ; Immunomagnetic Separation ; methods ; Interleukin-2 Receptor alpha Subunit ; metabolism ; Lymphocyte Count ; Mice ; Mice, Inbred C57BL ; T-Lymphocytes, Regulatory ; cytology

Objective To study the prevalence,risk factors of overactive bladder (OAB) in middle-aged and senior residents in Zhengzhou China.Methods A randomized,community-based,crosssectional study was performed on 10 160 residents aged 40 or older in urban area of Zhengzhou by using a stratified system sampling approach.A questionnaire including the subjects' basic information,previous history,present history,the Chinese overactive bladder symptom score (OABSS) was filled on site.The diagnostic criteria for OAB was 'an urgency score for Question 3 of 2 or more,and an OABSS of 3 or more'.Chisquare test was used to determine the differences of prevalence between genders,age groups,BMI and people with and without diabetes mellitus (DM).A pairwise comparison was conducted between different age,BMI group by using Bonferroni method.Results A total of 10 160 residents were investigated and finally 9805 (96.5％) were qualified for final statistical analysis.The mean age was (57.9 ± 9.7) years.The overall prevalence of OAB was 2.1％ (209/9805),of which,with OABdry 1.0％,and OABwet 1.1％.Male subjects were more likely suffered from OAB than female,with 2.7％ (84/3129) versus 1.9％ (125/6676).The prevalence of OAB in both male and female increased with age.There was no significant difference in the prevalence of male and female before the age of 60 years (1.2％ versus 1.4％,P ＞ 0.05) and more common in men than in women after the age of 60 years (4.6％ versus 2.6％,P ＜ 0.05).The prevalence of the subjects with DM was significantly higher than those without DM (P ＜ 0.05).The subjects with BMIs of 30 or more were nore likely to have OAB (3.2％ versus 1.8％,P ＜ 0.05).Conclusions The prevalence of OAB increases with advancing age.The prevalence of male is higher than female after the age of 60 years.The diabetics and obese people are more likely to have OAB.

Animals ; Dose-Response Relationship, Drug ; Guinea Pigs ; Heart Ventricles ; Ion Channels ; antagonists & inhibitors ; physiology ; Male ; Myocytes, Cardiac ; drug effects ; physiology ; Promethazine ; pharmacology ; Terfenadine ; pharmacology ; Time Factors

OBJECTIVEThe aim of this study was to explore the possibility of creating a toxin, C-CPE-ETA', by fusing C-terminal high affinity binding domain of CPE (C-CPE) with a truncated form of Pseudomonas aeruginosa exotoxin A (ETA') and to examine whether C-CPE-ETA' could specifically target CLDN-3, 4 molecule and the targeted toxin was cytotoxic against CLDN-3,4-overexpressing ovarian cancer.

METHODSCLDN-3 and CLDN-4 expressions were analyzed at the mRNA level in three ovarian cancer cell lines and epithelial ovarian cancer tissues from 20 patients. After transforming an expression plasmid of C-CPE-ETA' into E. coli BL21 (DE3) plysS strain, the recombinant protein was purified using His-Bind resin chromatography column and analyzed by Western blot and Coomassie blue staining. The specific binding, proapoptotic and cytolytic activities were evaluated by flow cytometry, fluorescence microscopy with the JC-1 probe and MTT assay in CLDN-3,4-overexpressing ovarian cancer cells.

RESULTSQuantitive RT-PCR results showed there existed high levels of CLDN-3 and CLDN-4 in ovarian cancer cells, CAOV3, OVCAR3 and SKOV3. Moreover, high expressions of CLDN-3 and CLDN-4 were observed in 90.0% (18/20) and 60.0% (12/20) of ovarian cancer tissues, with an expression level 10-fold higher than that in the normal ovarian tissue. A 58 000 recombinant protein C-CPE-ETA' was demonstrated by Western blot and Coomassie blue staining. Purified and recombinant C-CPE-ETA' was bound with high affinity to CLDN-3,4-overexpressing ovarian cancer cells, CAOV3, OVCAR3 and SKOV3 cells. C-CPE-ETA' was strongly proapoptotic and cytotoxic towards the CLDN-3,4-overexpressing ovarian cancer cells. The concentration of IC(50) was 7.364 ng/ml for CAOV3 cells, 8.110 ng/ml for OVCAR3 cells and 22.340 ng/ml for SKOV3 cells, respectively. However, control CLDN-3,4-deficient cell line HUVEC was not susceptible to the recombinant C-CPE-ETA' at a concentration up to 10 µg/ml.

CONCLUSIONSThe C-CPE-ETA' protein exhibits remarkably specific cytotoxicity for CLDN-3,4-overexpressing ovarian cancer cells. Its therapeutic potential warrants further development for ovarian cancer molecular targeted therapy.

ADP Ribose Transferases ; metabolism ; physiology ; Apoptosis ; Bacterial Toxins ; metabolism ; Cell Line, Tumor ; Claudin-3 ; Claudin-4 ; Claudins ; genetics ; metabolism ; Enterotoxins ; metabolism ; physiology ; Exotoxins ; metabolism ; physiology ; Female ; Humans ; Immunotoxins ; metabolism ; Ovarian Neoplasms ; metabolism ; pathology ; RNA, Messenger ; metabolism ; Recombinant Fusion Proteins ; metabolism ; physiology ; Virulence Factors ; metabolism ; physiology