Animals ; Antigens, CD ; immunology ; Cytokines ; metabolism ; Hepacivirus ; immunology ; Hepatitis C ; immunology ; prevention & control ; virology ; Hepatitis C Antibodies ; biosynthesis ; immunology ; Hepatitis C Antigens ; immunology ; Humans ; Killer Cells, Natural ; immunology ; T-Lymphocytes ; immunology ; metabolism ; T-Lymphocytes, Regulatory ; immunology ; metabolism ; Viral Proteins ; immunology

BACKGROUNDWilms' tumor (nephroblastoma) is a cancer of the kidneys that occurs typically in children and rarely in adults. Early diagnosis is very important for the treatment and prognosis of the disease. The aim of our study was to discover and identify potential non-invasive and convenient biomarkers for the diagnosis of Wilms' tumor.

METHODSNude mice were used to construct a Wilms' tumor model by injecting nephroblastoma cells into their bilateral abdomen. We collected 94 serum samples from mice consisting of 45 samples with Wilms' tumor and 49 controls. The serum proteomic profiles of the samples were analyzed via surface-enhanced laser desorption/ionization time-of-flight mass spectrometry. The candidate biomarkers were purified by high-performance liquid chromatography, identified by liquid chromatography-mass spectrometry, and validated using ProteinChip immunoassays.

RESULTSWe finally retrieved two differential proteins (m/z 4509.2; 6207.9), which were identified as apolipoprotein A-II and polyubiquitin, respectively. The expression of apolipoprotein A-II was higher in the Wilms' tumor group than in the control group (P < 0.01). By contrast, the expression of polyubiquitin was lower in the Wilms' tumor group than in the control group.

CONCLUSIONApolipoprotein A-II and polyubiquitin may be used as potential biomarkers for nephroblastoma in children, and the analysis of apolipoprotein A-II may help diagnose and treat Wilms' tumor.

Animals ; Apolipoprotein A-II ; blood ; Biomarkers ; blood ; Cell Line, Tumor ; Mice ; Mice, Nude ; Polyubiquitin ; blood ; Proteomics ; methods ; Wilms Tumor ; blood ; metabolism ; pathology

OBJECTIVESTo construct and screen a primarily phage display library of HCV C and E1 genes evolved with an artificial pattern.

METHODSTwo genes of about 1 kb with different genotypes were evolved by DNA shuffling. The re-assembled HCV C and E1 genes were cloned into a phage vector. After being rescued with helper phage M13KO7, a phage display library was constructed. Then the library was screened with anti-C and E1 McAb. Double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) was carried out on twenty individual phage clones selected randomly to detect their binding and reactive activity with high-titer HCV-positive sera. Normal sera were used as controls.

RESULTSThe phage display library of HCV C and E1 genes which evolved with an artificial pattern was constructed. Their capacity amounted to 1.64 x 10(6), and 86 percent of the clones contained C and E1 genes. After four rounds of panning, the phage library was specifically enriched. Twelve positive clones were successfully screened.

CONCLUSIONThe capacity and diversity of the constructed library are enough for screening. The results demonstrate the superiority of the specific binding and reactive activity and affinity of the 12 phage clones from the HCV positive sera.

DNA, Viral ; genetics ; Gene Library ; Hepacivirus ; genetics ; Peptide Library ; Viral Core Proteins ; genetics ; Viral Envelope Proteins ; genetics

OBJECTIVETo generate hepatitis C virus pseudo-particles (HCVpp) containing the complete E1-E2 envelope glycoprotein, in order to establish a HCVpp database covering the six major genotypes of HCV (1b, 2a, 3b, 4, 5, and 6) and to develop a simple and effective method for detection of neutralizing antibodies in HCV patients.

METHODSHCVpp were generated for the six genotypes by co-transfecting 293T cells with a plasmid expressing the respective E1-E2 (p HR, CMVA 8.2 construct) and a MLV-GFP plasmid. Titration of each HCVpp was carried out by p24 ELISA. Infectivity of each HCVpp was assessed by mixing the harvested supernatant of producer cells with sera from HCV patients, adding the mixture to Huh-7 cells, and detecting the subsequent titers of neutralizing antibodies against HCVpp.

RESULTSAll six types of HCVpp were able to infect Huh-7 cells in vitro. For healthy HCV carriers, only two genotypes of HCVpp (1b and 2a) produced neutralizing antibody titers more than 1:40. For cured HCV patients, only the 1b genotype produced neutralizing antibody titers more than 1:40. One patient showed titer of 1:200 for genotype 4. A healthy spouse of a chronic hepatitis C patient showed titers more than 1:40 for four genotypes of HCVpp (3a, 4, 5, 6).

CONCLUSIONWe generated six different genotypes of HCVpp successfully, established the in vitro neutralizing antibody detection method, and provided an effective model for screening antiviral drugs.

Adolescent ; Adult ; Antibodies, Neutralizing ; blood ; Antibodies, Viral ; blood ; Female ; Genotype ; Hepacivirus ; classification ; Hepatitis C ; blood ; immunology ; Humans ; Male ; Middle Aged ; RNA, Viral ; blood ; Viral Envelope Proteins ; immunology ; Young Adult

OBJECTIVETo discuss the specificity of diseases treated by moxibustion and fire needling in clinical practice, so as to provide references for clinical treatment.

METHODSWith data mining of modern computer technique, journal and literature databases regarding moxibustion and fire needling were established, respectively. Literature regarding moxibustion and fire needling for the past 60 years has been collected, screened, included, reviewed and abstracted. The utility rate of moxibustion and fire needling in each department was calculated, frequency of diseases in clinical practice was summarized, and diseases which had differences in clinical practice in each department were screened; also the advantages of disease categories and clinical practice between two kinds of therapies in each department were compared.

RESULTS(1) The utility rate of moxibustion was highest in department of internal medicine and surgery, which were 43.6% and 28.1%, respectively; the utility rate of fire needling was highest in surgery and dermatological department, which were 53.7% and 23.8%, respectively. (2) According to the comparison and analysis on diseases treated by two therapies in clinic, among 26 kinds of gynecology diseases that were treated by moxibustion, 20 kinds were not involved with fire needling; among 22 kinds of pediatrics diseases that were treated by moxibustion, 20 kinds were not involved with fire needling. It was certain that the difference of the two therapies in clinical application was more significant in gynecology and pediatrics than that in the rest four departments. (3) Among the diseases which had differences in clinical practice in each department, the ones involved with moxibustion alone were insomnia, distention and fullness, consumptive fatigue in the department of internal medicine, blood-vessel Bi, stiff neck and hernia in surgery department, urticarial, skin Bi and skin cancer in dermatological department, malposition, infertility and amenorrhea in gynecology department, diarrhea, indigestion and stomachache in pediatrics department, blepharoptosis, blurred vision and dryness syndrome in ENT department; the ones involved with fire needling alone were numbness, coldness syndrome and acute renal colic in the department of internal medicine, lipoma, soft tissue injury and papilloma in surgery department, bromhidrosis, freckle and erysipelas in dermatological department, uterine fibroid in gynecology department, umbilical polyp in pediatrics department, auricle pseudocyst, starred nebula and phlegmatic mass in ENT department.

CONCLUSIONMoxibustion is frequently applied in department of internal medicine and surgery, while fire needling is frequently used in surgery and dermatological department; the application of moxibustion is broader than that of fire needling in gynecology and pediatrics department. Among the diseases which have differences in clinical practice in each department, moxibustion is commonly seen for clinical symptoms featured with deficiency syndrome, while fire needling is commonly applied for the diseases that have obvious local symptoms.

Acupuncture Therapy ; instrumentation ; methods ; Clinical Trials as Topic ; Data Mining ; Humans ; Moxibustion ; instrumentation ; methods

AIMTo assay the transcriptional regulation effect of hHSF1 on prohibitin gene promoter.

METHODSThe total length of hHSF1 exon was amplified by PCR method and cloned into pcDNA3. 1(+) vector. pcDNA3. 1(+)-hHSF1 and pGI3 prohibitin were co-transfected into HEK293 cells. The luciferase activity was detected by Dual-Luciferase Reporter Assay System to evaluate the transcriptional regulation effect of hHSF1 on prohibitin gene promoter.

RESULTSThe pcDNA3.1(+)-hHSF1 vector was constructed successfully. The assay of luciferase activity showed that the transcription of pGL3-prohibitin was dramatically upregulated by hHSF1.

CONCLUSIONhHSF1 can transcriptionally regulate prohibitin expression.

Base Sequence ; DNA-Binding Proteins ; physiology ; Gene Expression Regulation ; HEK293 Cells ; Heat Shock Transcription Factors ; Humans ; Molecular Sequence Data ; Promoter Regions, Genetic ; Repressor Proteins ; genetics ; Transcription Factors ; physiology ; Transcription, Genetic ; Transfection

OBJECTIVESTo obtain prokaryotic expressed IA-2 recombinant protein and to identify its immunological activity.

METHODSThe complimentary DNA (cDNA) coding for the intracytoplasmic part of IA-2 (IA-2ic) was amplified from human fetal brain RNA, and was subcloned into the PinPoint Xa-1 T vector to construct recombinant expression plasmid, and was then expressed in E. coli JM109 cells as a fusion protein with a biotinylated peptide sequence at the aminoterminus. The biotinylated fusion protein was then purified by affinity chromatography and was subsequently dialyzed. Finally, its immunogenicity was evaluated by enzyme linked immunosorbent assay (ELISA).

RESULTSThe purified IA-2ic fusion protein resolved on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) as a single Coomassie brilliant blue stained band with a molecular weight of 59 kDa and its immunogenicity was confirmed by ELISA.

CONCLUSIONSE. coli expressed IA-2ic fusion protein has immunological activity. It can be used for detection of IA-2 autoantibodies (IA-2A) and for further studies on type 1 diabetes in future.

Animals ; Autoantigens ; biosynthesis ; DNA, Complementary ; analysis ; Diabetes Mellitus, Type 1 ; immunology ; Escherichia coli ; genetics ; Humans ; Membrane Proteins ; biosynthesis ; genetics ; isolation & purification ; Plasmids ; Protein Tyrosine Phosphatase, Non-Receptor Type 1 ; Protein Tyrosine Phosphatases ; biosynthesis ; genetics ; isolation & purification ; Rabbits ; Receptor-Like Protein Tyrosine Phosphatases, Class 8 ; Recombinant Fusion Proteins ; biosynthesis ; immunology ; isolation & purification

OBJECTIVETo study the clinical characteristics of pediatric hemorrhagic fever with renal syndrome (HFRS), and to improve its understanding so as to reduce the misdiagnosis.

METHODSA retrospective analysis was performed on the clinical data of 26 children with HFRS between January 2009 and December 2012.

RESULTSThe age of disease onset was mainly distributed between 7 and 14 years (23 cases, 88%), and the male-to-female ratio was 1.89:l. The clinical manifestations of pediatric HFRS varied. The early symptoms resembled those of a cold, and in the course of HFRS, most patients developed digestive symptoms such as vomiting and abdominal pain. The laboratory examinations usually implicated platelet changes, and the imaging examinations revealed polyserous effusions. The prominent complication was myocardial injury.

CONCLUSIONSPediatric HFRS mainly occurs in school-age children, more commonly in males. HFRS does not have typical clinical manifestations or symptoms, so it should be distinguished from cold or appendicitis at the early stage. When applying the fluid replacement therapy, the cardiac function should be carefully monitored in case of heart failure.

Adolescent ; Child ; Child, Preschool ; Female ; Fluid Therapy ; Hemorrhagic Fever with Renal Syndrome ; diagnosis ; therapy ; Humans ; Male ; Retrospective Studies

Amino Acid Sequence ; Epitopes, T-Lymphocyte ; immunology ; Forecasting ; HLA Antigens ; immunology ; Hepacivirus ; genetics ; immunology ; Hepatitis C ; immunology ; virology ; Hepatitis C Antigens ; immunology ; Humans ; T-Lymphocytes, Cytotoxic ; immunology ; virology ; Viral Hepatitis Vaccines ; immunology

To investigate the antiviral efficacy of combination therapy with pegylated-interferon alpha (peg-IFNa)-2a and ribavirin (RBV) in hepatitis C patients with liver cirrhosis after splenectomy or partial splenic embolization. Forty-nine hepatitis C patients with liver cirrhosis who were unable to use antiviral therapy because of hypersplenism were recruited for study and treated with splenectomy or partial splenic embolization. Three months later, a regimen of antiviral combination therapy was initiated with peg-IFNa-2a (once-weekly subcutaneous injection: 135 μg or 180 μg) and RBV (daily oral: 800 to 1200 mg), and was maintained for 48 weeks. The patients were followed up at treatment weeks 1, 2, 4, 6, 8, and 12. Thereafter, follow-up was conducted every four weeks. The patients were observed until 24 weeks after treatment discontinuation. Follow-up testing included liver function, blood chemistry, renal function, and HCV RNA level. Any adverse reactions were recorded. Liver cirrhosis patients complicated by hypersplenism can be treated effectively with peg-IFNa-2a/RBV combination antiviral therapy after splenectomy or partial splenic embolization. The antiviral-induced sustained viral response rates was 65.00% in cirrhotic/hypersplenic hepatitis C patients receiving splenectomy and 58.62% in those receiving partial splenic embolization. Hypersplenism patients with hepatitis C-related cirrhosis achieved a good antiviral therapeutic effect with peg-IFNa-2a/RBV combination therapy following splenectomy or partial splenic embolization. This sequence of treatment may help to decrease incidences of chronic hepatitis C-induced liver failure and liver cancer in these patients.
Adult ; Aged ; Antiviral Agents ; therapeutic use ; Combined Modality Therapy ; Female ; Hepatitis C ; complications ; therapy ; Humans ; Interferon-alpha ; therapeutic use ; Liver Cirrhosis ; etiology ; therapy ; Male ; Middle Aged ; Polyethylene Glycols ; therapeutic use ; Postoperative Period ; Recombinant Proteins ; therapeutic use ; Ribavirin ; therapeutic use ; Splenectomy ; Treatment Outcome