Objective To investigate the clinical value of sB7-H3 in predicting the severity of acute pancreatitis at early stage. Methods By using the double antibody sandwich ELISA method, the level of plasma sB7-H3 was measured at 24h after onset of abdominal pain in 75 patients with acute pancreatitis (MAP30, MSAP20, SAP25), and 20 healthy persons were enrolled in the controlgroup.The sensitivity and specificity correlations of sB7-H3 in acute pancreatitis with severity degree , as well as with the clinical detection index , were also evaluated. Results The level of plasma sB7-H3 at 24 h in the AP group was significantly higher than that in the healthy control ( HC) group (t = 3.925, P = 0.0002), however, no significant difference was found between the MAP groep and the HC group (P>0.05). The levels of plasma sB7-H3 in the MSAP and the SAP group were significantly higher than that in the HC group (P<0.05和P<0.001)or the MAP group (P<0.05 和P<0.01);The level of plasma sB7-H3 in the SAP group was also markedly higher than that in the MSAP group (P < 0.01). sB7-H3 had a linear positive correlation with LDH、hs-CRP、WBC(P<0.05). ALB had a linear negative correlation with and Ca (S)(P<0.05). By the cutoff of sB7-H3, the sensitivity and specificity to judgethe above moderate pancreatitis were 88.9%and 83.3%,and to judgethe SAP were 96%and 96%. Conclusion sB7-H3 has important clinical value to judge the severity of acute pancreatitis at early time with high sensitivity and specificity , with a linear correlation with the clinical severity index.

Objective To prepare a peptide monoclonal antibody (McAb)against human papillomavirus 18E6 separately,and identify its specificity and pathogenicity.Metheds The advantage epitope peptide was designed and synthesized by ABCpred and Bcepred,and then used to immunize BALB/c mice after coupling with bovine serum albumin (BSA).And the McAb was prepared by hybridoma technique.HPV18E6 gene was amplified from cervical swab specimen containing HPV18 and insert-ed into expression vector pET-28a.The constructed recombinant plasmid was transformed to E.coli BL21(DE3)for expres-sion under induction of isopropyl thio-β-D-galactoside.The expressed protein was used to identified the McAb had been pre-pared.Results The hybridoma cell lines could constantly produce MAbs against HPV18E6 peptides.Sequencing proved that recombinant plasmid pET-28a-HPV18E6 was constructed correctly.Western blotting showed that the anti-HPV18E6 pep-tides antibody could specifically recognize HPV18E6.Conclusion A monoclonal antibody against the advantage epitope pep-tide of human papillomavirus 18E6 prepared could specifically recognize HPV18E6 specifically.

Objective To detect serum biomarkers for pancreatic cancer associated diabetes and establish a model for diagnosis.Methods SELDI-TOF-MS was used to detect the differentially expressed serum proteins from 17 pancreatic cancer associated diabetes patients,17 new-onset type Ⅱ diabetes patients and 17 healthy controls,then a model of biomarkers was constructed and validated by Biomarker Patterns Software 5.0.Results Twelve discriminating m/z peaks were identified in the protein fingerprints in 10 pancreatic cancer associated diabetes patients,10 new-onset type Ⅱ diabetes patients and 10 healthy controls.Among them,the three biomarkers of mass/charge ratio 6116,6695 and 8936 were used to construct the model,which could diagnose 90％ pancreatic cancer associated diabetes form control groups.Blind test of other7 samples of three groups showed that 100％ pancreatic cancer associated diabetes,71％ new-onset diabetes and 86％ healthy controls were correctly classified.After searching protein database,there were metallothionein,pancreatic progenitor cell differentiation and proliferation factor-like protein,and fibroblastic growth factor 1,which were close to the weights of the above mentioned 3 differentially expressed proteins.Conclusions SELDI can identify 3 biomarkers for pancreatic cancer associated diabetes and a reliable model for diagnosis of pancreatic cancer associated diabetes is established.

OBJECTIVETo test whether splicing overlapping extension(SOE) method can be a tool for obtaining rare fusion gene's transcripts and to study the tumorigenic capacity of a novel fusion gene AML1-MTG16.

METHODSSOE method was used to obtain AML1- MTG16 fusion gene's transcripts. MTG16, AML1-MTG16 and AML1-MTG16 without III,VI conserved domains of MTG16 segment were inserted into pEGFP- C1,pDsRed-N1 vector respectively,then transfected NIH3T3 cell line by lipofection. Forty-eight hours later, the transfected cells were examined by laser-scanning confocal microscopy. Stable transfected cells were obtained by G418 500ug/ul selection for one month. Growth curve, soft agar colonies formation tumorigenesis in nude mice were done to compare the difference between stable transfected cells.

RESULTSRecombined AML1-MTG16 by SOE contained its CDS. NIH3T3 expressing AML1-MTG16 had a faster proliferation in medium, colony growth in soft agar. AML1-MTG16 expression cells also induced tumors formation following injection into nude mouse. MTG16,AML1-MTG16 and AML1-MTG16 without III,VI conserved domains of MTG16 were colocalized in the nucleus of cotransfected NIH3T3 cells under the examination of laser-scanning confocal microscope.

CONCLUSIONSOE is an effective method to get rare fusion gene's transcripts. AML1-MTG16 plays an important role in leukemogenesis. MTG16 may also have a carcinogenic property within the AML1-MTG16 fusion gene. Carcinogenic property of AML1-MTG16 is restricted to its localization in the nuclear matrix. N terminal of MTG16 may play an important part in the carcinogenic activity of AML1-MTG16.

3T3 Cells ; transplantation ; Animals ; Cell Division ; genetics ; Cell Transformation, Neoplastic ; genetics ; Cell Transplantation ; Core Binding Factor Alpha 2 Subunit ; Green Fluorescent Proteins ; Luminescent Proteins ; genetics ; metabolism ; Mice ; Mice, Nude ; Microscopy, Confocal ; Neoplasms, Experimental ; genetics ; pathology ; Oncogene Proteins, Fusion ; genetics ; Plasmids ; genetics ; Recombinant Fusion Proteins ; genetics ; metabolism ; Time Factors ; Transcription Factors ; genetics ; Transfection

OBJECTIVE: To clarify the association between regulatory region of HLA-DPB1 (3'UTR) with Naso pharyngeal carcinoma in Guangdong Province Hans. METHOD: The allelic types of HLA-DPB1-3'UTR were detected by sequence specific primer (SSP) in 104 patients with NPC and 105 control individuals from Guangdong Province Hans. RESULT: The frequencies of allelic types B/B, haplotype B were higher in patients with NPC than those of the control individuals. CONCLUSION Positive association may exist between certain HLA-DPB1 alleles and NPC in Guangdong Province Hans.
Alleles ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; Case-Control Studies ; Gene Frequency ; HLA-DP Antigens ; genetics ; HLA-DP beta-Chains ; Haplotypes ; Humans ; Molecular Sequence Data ; Nasopharyngeal Neoplasms ; genetics ; Polymorphism, Single Nucleotide

ObjectiveTo investigate the therapeutic effect of Qingmei compound on acute gouty arthritis （AGA） in rats and preliminarily clarify its mechanism. MethodForty male SD rats were randomly divided into a blank group， a model group， a colchicine group （0.3 mg·kg^-1）， and low- and high-dose Qingmei compound groups （200 and 400 mg·kg^-1）， with eight rats in each group. The AGA model was induced by injecting 50 g·L^-1 monosodium urate （MSU） into the ankle joint of the rats except those in the blank group. The ankle swelling index was measured before and 6， 24， and 48 h after modeling. The pathological changes in the joint tissues of AGA rats were observed by hematoxylin-eosin （HE） staining. The expression of tumor necrosis factor-α （TNF-α） and interleukin-1β （IL-1β） in the joint tissues of rats was detected by immunohistochemistry. The protein expression of NOD-like receptor protein 3 （NLRP3） pathway and key proteins in the joint tissues of rats was detected by Western blot. ResultCompared with the blank group， the model group showed increased ankle swelling index， synovial hyperplasia， and inflammatory infiltration， and up-regulated expression of IL-1β， TNF-α， and NLRP3 proteins in the ankle joint and the ratio of Caspase-1 shear body to Caspase-1 precursor protein （Caspase-1 p20/Caspase-1） （P<0.01）. Compared with the model group， the Qingmei compound groups showed reduced ankle swelling index of AGA rats， especially the low-dose Qingmei compound group （P<0.01）. Meanwhile， Qingmei compound inhibited synovial hyperplasia and inflammatory infiltration （P<0.01） and reduced the levels of IL-1β， TNF-α， and NLRP3 proteins and Caspase-1 p20/Caspase-1 in joint tissues （P<0.01）. ConclusionQingmei Compound can significantly alleviate the joint swelling and inflammatory infiltration of AGA， and its mechanism may be related to the inhibition of the NLRP3 signaling pathway.