To identify SJCHGC01743 gene of Schistosoma japonicum and evaluate the potential of the recombinant protein as a new vaccine candidate for schistosomiasis, polymerase chain reaction (PCR) technique was used to amplify the cDNA of the gene and real-time RT-PCR was used to analyze the transcription profiles of SJCHGC01743 at different development stages. Recombinant plasmid was successfully constructed and transformed into competent Escherichia coli BL21 (DE3). Then the recombinant protein was expressed, purified and emulsified with ISA206 adjuvant to immunize BALB/c mice for three times. The immunogenicity was confirmed by Western blotting and tissue localization was detected by indirect immunofluorescent assay. The specific antibody level was detected by ELISA. The immunoprotection of rSjOST48 was evaluated by the reduction in worm and egg counts in mice. A cDNA with 1 248 nucleotides was isolated from 28-day-old schistosomes cDNAs by PCR. Sequence analysis revealed that SJCHGC01743 was a 48-kDa subunit of the oligosaccharyltransferase complex (OST48) and named as SjOST48. Real-time PCR analysis indicated that this gene was expressed in all investigated stages and had the highest expression level in 28 d worms, the level of gene transcription in female worms was significantly higher than that of male worms. Then recombinant plasmid pET28a(+)-SjOST48 was successfully constructed and expressed in E. coli BL21 (DE3). Western blotting analysis showed that rSjOST48 had good immunogenicity. Indirect immunofluorescent analysis revealed that SjOST48 was mainly distributed on the tegument of the worms. The result of ELISA indicated that the rSjOST48 vaccinated group could induce a significant increase in the level of specific IgG, IgG1 and IgG2a. An immunoprotection experiment showed that the vaccination of rSjOST48 in mice induced 32.62% (P < 0.05) reduction in the numbers of worms and 57.61% (P < 0.01) in eggs in liver, compared with that of the control group. This study provides the foundation for proceeding further research on the biological function of SjOST48 and screening new vaccine candidates for schistosomiasis.
Animals ; Antibodies, Helminth ; blood ; Cloning, Molecular ; DNA, Complementary ; Escherichia coli ; Female ; Genes, Helminth ; Helminth Proteins ; genetics ; immunology ; Immunoglobulin G ; blood ; Male ; Mice ; Mice, Inbred BALB C ; Real-Time Polymerase Chain Reaction ; Recombinant Proteins ; immunology ; Schistosoma japonicum ; genetics ; Schistosomiasis japonica ; prevention & control ; Vaccination

OBJECTIVETo investigate the differential diagnosis of type III prostatitis and interstitial cystitis so as to improve the efficiency of diagnosis and treatment of the two diseases.

METHODSBased on the clinical data of 4 cases of type III prostatitis and 3 cases of interstitial cystitis, we analyzed the characteristics of the two diseases in such aspects as clinical symptomatology, urodynamics, prostatic fluid microscopy, microbiology and treatment.

RESULTSThe common clinical characteristics of type III prostatitis and interstitial cystitis were indisposition or pain in the subabdomen and/or pelvic floor, but their differences were quite obvious. In interstitial cystitis, longer urine accumulation could cause worse pain in the subabdomen, which could be relieved after micturation, and the bladder capacity was obviously decreased, but with normal prostatic fluid and negative result of microbial culture. It responded to behavior therapy, resiniferatoxin, sodium hyaluronate and water dilation of the bladder under anaesthesia. While type III prostatitis, with white blood cells > 10/HP or < or = 10/HP in the prostatic fluid and negative result of microbial culture, did not respond to the above therapeutic methods that were effective for interstitial cystitis.

CONCLUSIONType III prostatitis and interstitial cystitis, although clinically confusable, can be definitely differentiated from each other according to their characteristic causes and locations.

Adult ; Aged ; Cystitis, Interstitial ; diagnosis ; etiology ; Diagnosis, Differential ; Humans ; Male ; Middle Aged ; Prostatitis ; complications ; diagnosis

OBJECTIVETo study the possible mechanism of the intracellular aggregate formation of small heat shock protein HSPB8 (HSPB8)(K141N) mutation resulting in axonal Charcot-Marie-Tooth disease type 2L(CMT2L).

METHODSThe cell models which transiently expressed pEGFPN1-HSPB8 and pEGFPN1-(K141N)HSPB8 were established. The immunofluorescent co-location study of EGFP-(K141N)HSPB8 and HSPB1, EGFP-(K141N)HSPB8 and neurofilament light chain (NEFL) was carried out in the SHSY5Y cell models. The aggregate formation of EGFP-(K141N)HSPB8 in cell models was investigated and the possible mechanism of cellular aggregate formation was analyzed by t test and analysis of variance between group(ANOVA).

RESULTSEGFP-(K141N)HSPB8 formed large aggregate which predominantly located around the nucleus in cell models. EGFP-(K141N)HSPB8 co-localized perfectly with HSPB1 and NEFL in the SHSY5Y cell models. The aggregate formation was different in different cell types, there were fewer aggregates formed in an sHSPs deficient milieu than in HEK293T cells.

CONCLUSION(K141N)HSPB8 formed aggregates predominantly locate around the nucleus in cells. (K141N)HSPB8 co-localizes perfectly with HSPB1 and NEFL. The aggregate formation may be due to (K141N)HSPB8 conformational change leading to self aggregation and its abnormal interaction with other sHSPs such as HSPB1.

Cell Line ; Cell Line, Tumor ; Cell Nucleus ; metabolism ; Charcot-Marie-Tooth Disease ; genetics ; metabolism ; Green Fluorescent Proteins ; genetics ; metabolism ; HSP27 Heat-Shock Proteins ; HeLa Cells ; Heat-Shock Proteins ; genetics ; metabolism ; Humans ; Kidney ; cytology ; metabolism ; Microscopy, Confocal ; Neoplasm Proteins ; genetics ; metabolism ; Neuroblastoma ; genetics ; metabolism ; pathology ; Neurofilament Proteins ; genetics ; metabolism ; Point Mutation ; Protein-Serine-Threonine Kinases ; genetics ; metabolism ; Recombinant Fusion Proteins ; genetics ; metabolism ; Transfection

BACKGROUNDEpstein-Barr virus (EBV) infection is one of the most important factors of nasopharyngeal carcinoma (NPC) endemic areas. Transcription of EBV-encoded non-polyadenylated RNAs (EBERs) are presented in most of NPC tumors. Exploring EBERs as a prognostic marker for NPC might further be informative about the biology and the progression of the disease. The aim of this study was to analyze the role of EBV latency in the clinical management of nasopharyngeal carcinoma (NPC), by detecting EBERs.

METHODSRNA in situ hybridization (ISH) for detecting EBERs was carried out on 908 NPC tumor tissues. Overall survival (OS) curves were analyzed with the Kaplan-Meier method and the Cox proportional-hazards regression models.

RESULTSThe median follow-up time was 70 months (1-120 months). Eight hundred and sixteen (89.9%) from a total of 908 consecutive NPC cases were found to be EBV-EBER positive. EBER-ISH staining revealed nuclear localization in NPC cells. In the Kaplan-Meier analysis for OS, high EBER expression levels in NPC patients were statistically significant positive prognostic factors for survival (log-rank, P = 0.022), especially in adults aged 17-40 years (P = 0.023) and in those with advanced stage disease (log-rank, P = 0.002). Cox proportional-hazards regression model analysis showed that the EBER expression level was an independent risk factor for OS (hazard ratio 0.724, P = 0.005).

CONCLUSIONSEBERs were frequently detected in NPC tumor tissues, and high-level EBER expression correlated with good prognosis in NPC patients, especially in adult patients and in those with advanced stage disease. EBER may serve as a potential prognostic predictor in NPC.

Adolescent ; Adult ; Aged ; Carcinoma ; Epstein-Barr Virus Infections ; virology ; Female ; Herpesvirus 4, Human ; genetics ; pathogenicity ; Humans ; In Situ Hybridization ; Male ; Middle Aged ; Nasopharyngeal Neoplasms ; mortality ; virology ; RNA, Viral ; genetics ; Young Adult