Objective To evaluate the application of western blot in treponema pallidum antibody screening among blood donors.Methods Through standard blood liquidation of syphilis,the sensitivity and specificity of two kinds of TP -ELISA regents and one TPPA regent were evaluated.TP -WB was used to test 279 positive specimens with TP -ELISA,and the correlation between test results of the two methods and the distribution characteristics of WB bands were analyzed. Results The sensitivity of the two kinds of TP -ELISA reagent was 1 00.00%,while the specificity were 92.86% and 85.71 % respectively.At the same,the sensitivity of TPPA reagent was 88.46%,while the specificity was 1 00.00%.In 279 positive specimens with TP -ELISA method,WB confirmed positive was 21 6,positive rate was 77.42%;Including S /Co value >5 and two kinds of ELISA reagent testing both positive specimens were 205,the WB confirmation test positive rate was 1 00.00%,accounted for 99.91 % in 21 6 WB positive samples.Orderly Logistic regression analysis,the method of ELISA S /Co value >5 and double reagent is positive,had statistical correlation with test positive for WB,respectively(P <0.01 );21 6 TP -WB positive specimens WB banding distribution analysis,TP1 7 belt with syphilis antibody IgM,TP1 5 belts and IgG +IgMand IgG exist statistical correlation respectively(P <0.01 ).Conclusion ELISA method of double reagent positive and S /Co value >5 specimens of the basic can be diagnosed with syphilis,WB test with positive results for blood donation member state judgment has guiding significance.

OBJECTIVETo evaluate the abnormal state of liver function and plasma lipid levels of obese schoolchildren who were screened by weight-for-height criterion and new body mass index criterion respectively.

METHODS280 obese children were screened by weight-for-height criterion and 125 obese children were screened by body mass index criterion in a routine school check-up program. All of the latter subjects was included in the former one. One obese child and 1 non-obese child were matched for gender and age. 14 items related to liver functions and plasma lipids were measured.

RESULTSOf the abnormal items,7 items in 125 obese children screened by new BMI criterion and 5 items in 155 "obese children" excluded by BMI criterion, were significantly higher than those children among controlled group. The abnormal rates were 10.4%-22.9% in the former and 3.2%-13.0% in the latter.

CONCLUSIONSThe new BMI criterion seemed to be more stringent than weight-for-height. Less than a half of the obese children screened by weight-for-height were taken on obese children by new BMI criterion. The overweight children who were screened by BMI criterion also had abnormal liver functions and plasma lipids.

Body Mass Index ; Case-Control Studies ; Child ; Humans ; Lipids ; blood ; Liver ; physiopathology ; Obesity ; blood ; physiopathology

OBJECTIVETo study the clinical characteristics of male urethral duplication infection and offer some guidelines for the diagnosis and treatment of the disease.

METHODSWe analyzed the pathological types, clinical characteristics, therapeutic processes and follow-up results of 9 cases of male urethral duplication.

RESULTSAmong the 9 cases of urethral duplication, 7 turned out to be of Type I, 1 Type II A2 and 1 Type II B. The disease courses varied from 2 to 420 days, with an average of 77.2 +/- 141.5 days. Four cases with longer disease duration were identified with a history of repeated use of various antibiotics for treatment. Their clinical manifestations varied, with the outflow of excretions or pus from the duplicate or normal urethra as the cardinal symptoms. The pathogens detected from the secretions were mainly Neisseria gonorrhoeae, Ureaplasma urealyticum, and Chlamydia trachomatis. The consistency rate of the same pathogens detected in the vaginal or cervical secretions from the sex partners of the patients was 87.5%. All the symptoms disappeared after a sufficient-course treatment with sensitive antibiotics, and the patients' sex partners received the same medication simultaneously. No recurrence was found during a 3-month follow-up.

CONCLUSIONUrethral duplication infection has various clinical manifestations, and thus is easily missed in diagnosis. Sufficient-course treatment with sensitive antibiotics is recommended for those that prefer conservative therapy, and their sex partners should be treated simultaneously.

Adult ; Humans ; Male ; Middle Aged ; Urethra ; microbiology ; Urethral Diseases ; diagnosis ; drug therapy ; microbiology ; Young Adult

To investigate the semi-quantitative method for evaluating lipid accumulation in livers, male C57BL/6J mice (CON), HFD mice characterized with the mild fatty liver induced by high-fat diet, and KKAy mice charactered with the moderately severe fatty liver induced by high-caloric diet were used. The lipid accumulation was estimated by the histological examination (HE staining) and the content of hepatic triglyceride, separately. Echo-intensity of two selected regions along the ultrasound transmission direction was recorded using a small animal ultrasonographic system, and the echo-intensity attenuation coefficient was calculated. Correlation between the echo-intensity attenuation coefficient and the content of hepatic triglyceride was analyzed by the Spearman's rank correlation analysis. The results showed that the lipid accumulation in livers increased significantly in both HFD and KKAy mice compared with CON mice and it was more serious in KKAy mice than that in HFD mice. The values of echo-intensity attenuation coefficient were also increased in sequence according to group. These values were positively associated with the content of hepatic triglyceride (r = 0.744, P < 0.01). In conclusion, the echo-intensity attenuation coefficient is a simple, impersonal, and non-invasive method for evaluating the hepatic lipid accumulation. It can be used to research the process and the treatment of fatty liver diseases in mice.
Animals ; Diet, High-Fat ; Fatty Liver ; diagnostic imaging ; Male ; Mice ; Mice, Inbred C57BL ; Triglycerides ; analysis ; Ultrasonography

OBJECTIVEMutation in the interleukin-7 receptor-alpha (IL-7R alpha) chain causes a rare type of severe combined immunodeficiency (SCID) with presence of NK cells in the peripheral blood. Here we report the molecular and clinical characterization of a compound heterozygosity mutation in the interleukin-7 receptor-alpha gene that resulted in SCID in a patient firstly from China.

METHODA 5 month-old male patient and his parents were enrolled in this study. Since 15 days of age, the patient had had recurrent fever, persistent cough and diarrhea. He was in poor general condition with pyorrhea and ulceration of the BCG scar. His brother died of severe infection at 4 months of age. He was initially diagnosed as SCID according to clinical manifestation and immunological analysis. A panel of SCID candidate genes including IL-2RG, RAG1/RAG2 and IL-7R alpha of patient and his parents were amplified by polymerase chain reaction (PCR) from genomic DNA. Reverse transcription polymerase chain reaction (RT-PCR) was used to amplify the IL-7R alpha transcripts. Sequencing was performed directly on the PCR products forward and reversely.

RESULTThe serum immunoglobulin (Ig) profile was IgG 6867 mg/L (normal range, 3050 - 8870 mg/L); IgM 206 mg/L and IgA 249 mg/L, IgE 2.3 IU/ml (normal range < 150 IU/ml). The patient was treated with IVIG previously. There were no T-cells but increased percentage of B-cells (58%) and NK cells (42%) in the peripheral blood was found. Needle biopsies from enlarged axillary lymph node was identified positive for Mycobacterium bovis under microscope and by culture. The patient had a compound heterozygosity mutation in the IL-7R alpha gene:on one allele, there was a splice-junction mutation in intron 4 (intron 4(+1)G > A), for which his father was a carrier; whereas on the other allele, a nonsense mutation at position 638 in exon 5 with a premature stop codon (638 C > T, R206X) was identified, for which his mother was a carrier. The splice-junction mutation in intron 4 of IL-7R alpha was firstly reported. The IL-7R alpha mRNA expression of the patient was remarkably reduced whereas the parents had relatively normal IL-7R alpha mRNA expression. IL-7R alpha cDNA of the patient was amplified by nested PCR. The PCR products were purified, cloned with a TA Cloning Kit and sequenced directly. A 64 bp deletion was found in exon 4 of IL-7R alpha. No mutation was found in IL-2RG and RAG1/RAG2 of the patient and his parents.

CONCLUSIONThis is the first case with a compound heterozygosity mutation in the IL-7R receptor alpha gene and T-B+NK+ phenotype from China. Intron 4(+1)G > A was a novel mutation.

DNA ; Genome, Human ; Heterozygote ; Humans ; Infant ; Male ; Mutation ; Receptors, Interleukin-7 ; genetics ; Severe Combined Immunodeficiency ; genetics

Adult ; Antigens, Neoplasm ; genetics ; Carcinoma, Hepatocellular ; genetics ; pathology ; Female ; Humans ; Liver ; pathology ; Liver Neoplasms ; genetics ; pathology ; Male ; Middle Aged ; RNA, Messenger ; genetics

OBJECTIVETo construct a prokaryotic expression vector for apoptin and prepare polyclonal antibody of apoptin.

METHODSApoptin gene amplified from pGEM-T/Apoptin plasmid by PCR was cloned into pET-28a (+). E.coli BL21 (DE3) was transformed by the recombinant plasmid, and apoptin protein expression induced by IPTG was analyzed by SDS-PAGE. BALB/c mice were immunized with the protein and the titer of the antibody was determined using indirect enzyme-linked immunosorbent assay (ELISA).

RESULTSApoptin gene was successfully cloned into pET-28a (+), and the expression of a protein with relative molecular mass of about 17 000 was identified by SDA-PAGE. After 5 immunizations of the mice with the protein, the blood antibody titer reached 1:5x10(5).

CONCLUSIONThe prokaryotic expression vector for apoptin is successfully constructed and the polyclonal antibody of apoptin is obtained, which allows further functional study of apoptin.

Animals ; Antibodies ; blood ; genetics ; immunology ; Capsid Proteins ; genetics ; immunology ; Cloning, Molecular ; Escherichia coli ; metabolism ; Gene Expression ; Genetic Vectors ; Genome ; Mice ; Mice, Inbred BALB C ; Plasmids

Aim To explore the apoptosis of small eell lung eancer ( SCLC ) eells HI688 and H446 induced by nitidine chloride and its possible mechanism.Methods The effect of nitidine chloride or cisplatin ( DDP ) on the activity of SCLC cells was detected by j J MTT method; the morphological changes of cells trea¬ted with nitidine chloride or DDP were observed by in- verted fluorescence microscope and HE staining; the effect of nitidine chloride or DDP on apoptosis was de¬tected by flow cytometry; the effect of apoptosis inhibi¬tor Z-VAD-FMK on apoptosis induced by nitidine chlo¬ride or DDP was detected by MTT method.The expres¬sions of Bax , Bcl-2, caspase-3 , PARP, p-PI3K and p- Akt in the cells treated with nitidine chloride or DDP were detected by Western blot.Results MTT results showed that the viability of SCLC cells was significantly reduced after 48 hours of treatment with nitidine chlo¬ ride; compared with DDP, nitidine chloride could in¬hibit SCLC cells with less IC50; inverted fluorescence microscope and HE staining showed that nitidine chlo¬ride could induce apoptosis in SCLC cells, similar to DDP; flow cytometry showed that nitidine chloride J J could induce apoptosis in SCLC cells.The results of MTT assay showed that the inhibitory effect of nitidine chloride on apoptosis of SCLC cells could be partially antagonized by apoptosis inhibitor Z-VAD-FMK.West¬ern blot results showed that, similar to DDP, nitidine chloride could inhibit the expression of PI3K and Akt, increase Bax, inhibit Be 1-2, and promote the cleavage of caspase-3 and PAH P.Conclusion Nitidine chlo¬ride can induce apoptosis of SCLC cells by inhibiting the activation of P13K and Akt.

OBJECTIVETo apply pulse-field gel electrophoresis analysis(PFGE) in the analysis of cholera outbreak events and to determine the molecular epidemiological characteristics of Vibrio cholerae ( V. cholerae) isolates.

METHODSPFGE using restriction enzyme Not I was employed in the molecular subtyping of forty-one strains of V. cholerae isolated in cholera outbreak events from 2003 to 2005 in Guangzhou area and PFGE patterns were analyzed by BioNumerics Version 4.0 software to perform cluster analysis. Pattern profiles were compared by utilizing of Dice coefficient and UPGMA(unweighted pair group method with arithmetic averages). Comparison of PFGE typing results was performed with phage-biological typing and pathogenicity-associated genes typing.

RESULTSIn cholera outbreak events, PFGE could discriminate epidemiologically related and unrelated strains, having more discriminatory power than phage-biological typing and pathogenicity-associated genes-typing.

CONCLUSIONSMolecular sub-typing by PFGE could disclose the epidemiological relationships of strains from humans and the environment, providing molecular epidemiological evidence and support for the source-tracking of cholera outbreak events.

Bacterial Typing Techniques ; methods ; Cholera ; epidemiology ; microbiology ; Disease Outbreaks ; Electrophoresis, Gel, Pulsed-Field ; Humans ; Molecular Epidemiology ; Vibrio cholerae ; classification ; genetics ; isolation & purification

To investigate the mechanisms of a compound (FF16), compatibility of Rhodiola crenulata, Cordyceps militaris, and Rheum palmatum, on glucose metabolic disorders, the IRF mice charactered with insulin resistance and glucose metabolic disorders induced by high-fat diet in C57BL/6J mice were randomly divided into 3 groups; IRF, rosiglitazone (Rosi) and FF16. The glucose metabolism was evaluated by fasting blood glucose (FBG) levels and intraperitoneal glucose tolerance test (IPGTT). The insulin sensitivity was estimated by insulin tolerance test (ITT), fasting serum insulin levels and the index of HOMA-IR. The expressions of Akt and its phosphorylation levels, GSK3beta and its phosphorylation levels in liver were detected by Western Blot. The results showed that FF16 significantly improved the glucose metabolic disorders through reducing FBG by 15.1%, decreasing AUC values in glucose tolerance tests by 22.3%. FF16 significantly improved the insulin sensitivity through decreasing AUC values in insulin tolerance tests by 22.1%, reducing the levels of serum insulin by 42.9% and of HOMA-IR by 49.5%, comparing with model control, respectively. After the treatment with FF16, the levels of p-Akt and p-GSK3beta were increased by 116.4% and 24.9%, respectively, in the liver of IRF mice. In conclusion, compound FF16 could improve glucose metabolic disorders in IRF mice through enhancing the glyconeogenesis.
Animals ; Cordyceps ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Glucose ; metabolism ; Glycogen Synthase Kinase 3 ; metabolism ; Glycogen Synthase Kinase 3 beta ; Insulin Resistance ; Male ; Metabolic Syndrome ; drug therapy ; Mice ; Mice, Inbred BALB C ; Phosphorylation ; Proto-Oncogene Proteins c-akt ; metabolism ; Rheum ; Rhodiola