Objective:To investigate the molecular interaction between non-structural protein 3 serine protease of hepatitis C virus(HCV)and wild type P53,and to lay the basis for elucidating the mechanism of oncogenesis of hepatocellular carcinoma(HCC)after infection of HCV.Methods:The recombinant plasmids,pGAD424-NS3,pGAD424-NS315aa- and pGAD424-NS330aa-,were constructed and the interaction between NS3 serine protease and its cofactor NS4A,the interaction between wild type P53 and NS3 serine protease and its N-truncated mutants were dectected qualitatively and quantitatively in yeast two-hybrid system.Results:The results indicated that interaction existed not only between full-length NS3 serine protease and P53,but also between N-truncated mutants of NS3 serine protease and P53.Furthermore,the difference between enzyme activity unit(IU)of β-gal induced by these interactions was not significant(P>0.05).Conclusions:NS3 serine protease of hepatitis C virus and its N-truncated mutants can interact with wild type P53,and the region of NS3 serine protease involved in the interaction may be located in its C-terminal,but not in its N-terminal.

Disease Outbreaks ; prevention & control ; Humans ; Influenza A Virus, H5N1 Subtype ; immunology ; Influenza Vaccines ; immunology ; Influenza, Human ; prevention & control ; Seasons ; Vaccination

Adenocarcinoma ; diagnosis ; metabolism ; pathology ; secondary ; Brain ; metabolism ; pathology ; Brain Neoplasms ; diagnosis ; metabolism ; pathology ; secondary ; Carcinoembryonic Antigen ; metabolism ; Diagnosis, Differential ; Female ; Humans ; Keratin-7 ; metabolism ; Lung Neoplasms ; pathology ; Magnetic Resonance Imaging ; Middle Aged ; Nuclear Proteins ; metabolism ; Thyroid Nuclear Factor 1 ; Transcription Factors ; metabolism

Objective：To investigate the correct expression of dengue 2 virus 43 strain NS1 gene in transfected BHK-21 cell. Methods：The D2-43 DNA fragment coding for signal peptide plus NS1 protein was cloned between KpnⅠ site and EcoR Ⅰ site of expression plamid pcDNA3.1. The obtained recombinant vector pcDNA-NS1 was transfected into BHK-21 cells with electroporation technique. After selection by G418, resistant clones were screened by RT-PCR and Western blotting test. Results：The RT-PCR results of four in five randomly selected cell clones were positive. Western blotting test showed that NS1 gene could be expressed in BHK-21 cells. Conclusions：NS1 protein was capable of being expressed and appropriately processed in pcDNA-NS1 transfected BHK-21 cells. The present results suggest the feasibility of NS1-based DNA immunization.

OBJECTIVEElectron microscopical study of infected cells to identify the pathogenic agent of SARS.

METHODSVero E6 cells infected with lung autopsy samples or nasopharyngeal swabs from SARS patients of Beijing and Guangzhou were inoculated. The supernatant and cultured cells exhibiting identifiable cytopathic effect (CPE) were prepared for electron microscopic study.

RESULTSExamination of CPE cells on thin-section revealed characteristic coronavirus particles within the cisternae of endoplasmic reticulum, Golgi apparatus, vesicles and extracellular space. They were mainly spherical or oval in shape, annular or dense, about 80 nm in diameter. Negative-stain electron microscopy identified coronavirus particles in culture supernatant, 80 - 120 nm in diameter, with club-shaped surface projections. Elongated, rod-, kidney- or other irregular shaped virons with the size of 100 - 200 nm by 60 - 90 nm were also found in the cultured cells infected with the lung samples from the Guangdong patients. Infectious virons entered cells by endocytosis or membrane fusion and released through a budding process.

CONCLUSIONThese data indicate a novel coronavirus as the causative agent of SARS. Most viral particles showed typical characteristics of coronavirus. The potential role of special shape viruses is expected to be further investigated.

Animals ; Cercopithecus aethiops ; Humans ; Microscopy, Electron ; SARS Virus ; ultrastructure ; Severe Acute Respiratory Syndrome ; virology ; Vero Cells

OBJECTIVETo generate rescued viruses with deletion mutation of capsid protein from dengue virus type 2 isolated in China (DEN2-43).

METHODSOn the basis of infectious full-length cDNA clone pD212 of DEN2-43 strain virus, the deletion mutants were constructed by fusion PCR, from which the rescued viruses with deletion mutation of capsid protein were generated by transcription in vitro and electroporation.

RESULT AND CONCLUSIONSequence analysis demonstrated that the deletion mutations had been successfully inserted into the rescued viruses obtained. These mutant viruses may hold the key for elucidating the effects of deletion mutation of capsid protein on the biological characteristics of dengue virus.

Amino Acid Sequence ; Animals ; Base Sequence ; Capsid Proteins ; genetics ; Cell Line ; Cloning, Molecular ; DNA Mutational Analysis ; Dengue Virus ; genetics ; isolation & purification ; Electroporation ; Molecular Sequence Data ; Sequence Deletion ; Sequence Homology, Nucleic Acid ; Transcription, Genetic ; Virus Replication ; genetics

OBJECTIVEScrub typhus is an infectious disease due to Orientia tsutsugamushi transmitted by infected chigger mites. Scrub typhus has long been recognized to occur in southern areas of China, but has recently been increasingly often reported from the north since the first case was reported in Mengyin County, Shandong Province in 1986. The key objectives of the present study were to investigate the clinical manifestations and epidemic factors of scrub typhus in children from the northern new natural foci.

METHODSThe case records of 56 children with scrub typhus who were admitted to the 5 hospitals of Fei County from September 1993 to January 2004 were reviewed. Orientia tsutsugamushi (Ot) was isolated from the cases. Based on ecological observations on the composition, seasonal fluctuation of animal hosts and chigger mites, Ot was isolated from rodents and chiggers. IgG antibodies to Ot was detected by IFA. Genotypes of the Ot isolates were also identified by nested PCR.

RESULTSAmong 56 children scrub typhus cases, 46 were male, 10 were female; 96% exhibited typical eschars or ulcers, 100% cases had high fever, skin rashes were observed in 55 cases (98%), and regional lymphadenopathy occurred in 48 cases (86%). All cases came from countryside, and all had histories of exposure to the crop field. fifty-one serum samples of suspected patients with scrub typhus were collected, 48 were positive for antibodies to Ot. The serotypes were Gilliam types. The cases only appeared in September to December with the peak at mid and late October. Leptotrombidium (L.) scutellare was the most important vector causing scrub typhus in the foci. Apodemus (A.) agrarius was the main host animals of Ot in the crop field. Totally 26 strains were isolated from patients, rodents, and chigger mites. The serotypes of 24 out of the 26 isolates were Gilliam types, while the genotypes of these isolates were Kawasaki types. The serotypes of the other 2 isolates were identical and both were Karp types.

CONCLUSIONChildren scrub typhus patients were frequently seen in the new natural foci of Shandong province. Exposure history, typical eschars or ulcers, and presence of IgG antibody were the important indexes to diagnose the disease.

Adolescent ; Animals ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Mice ; parasitology ; Orientia tsutsugamushi ; isolation & purification ; Scrub Typhus ; epidemiology ; Seasons ; Trombiculidae ; microbiology

OBJECTIVETo investigate the role of α-enolase (ENO1) in regulating glucose metabolism and cell growth in human glioma cells.

METHODSGlucose uptake and lactate generation were assessed to evaluate the changes in glucose metabolism in human glioma U251 cells with small interfering RNA (siRNA)-mediated ENO1 knockdown. MTT assay and 5-ethynyl-2'-deoxyuridine (EdU) staining were used to examine the cell growth and cell cycle changes following siRNA transfection of the cells.

RESULTSTransfection of U251 cells with siRNA-ENO1 markedly reduced glucose uptake (P=0.023) and lactate generation (P=0.007) in the cells and resulted in significant suppression of cell proliferation (*P<0.05) since the second day following the transfection. Transfection with siRNA-ENO1 also obviously suppressed cell cycle G1/S transition in the cells (P=0.0425). The expressions of HK2 and LDHA, the marker genes for glucose metabolism, were significantly down-regulated in the cells with siRNA-mediated ENO1 knockdown.

CONCLUSIONENO1 as a potential oncogene promotes glioma cell growth by positively modulating glucose metabolism.

OBJECTIVE: To assess changes of nutritional status by comprehensive nutrition assessment including nutritional risk screening, dietary assessment, blood biochemical index, and body composition in acute leukemia patients who had undergone chemotherapy. METHODS: A total of 169 patients with acute leukemia treated at The First Affiliated Hospital of Soochow University from June 2018 to August 2019 were recruited for this study. Before and after chemotherapy, the NRS-2002 and PG-SGA scales, dietary intake, blood biochemical index and body composition were evaluated to compare the changes of nutritional status. RESULTS: NRS-2002 score and PG-SGA score after chemotherapy were significantly increased than those before chemotherapy (P<0.001). Many patients had insufficient nutritional intake during chemotherapy, and the dietary intake score of patients with induction chemotherapy was significantly lower than that of patients with consolidation chemotherapy (P=0.043). The results of multivariate analysis showed that induction chemotherapy was the independent risk factor for the increase of PG-SGA scores and the decrease of dietary intake (all P<0.05). After chemotherapy, the white blood cell count, hemoglobin, and platelet count were significantly decreased (P<0.001), the prealbumin was significantly increased (P<0.001), and the blood glucose was increased (P=0.04), but albumin was not significantly changed. The weight, body mass index, fat-free mass, skeletal muscle mass and intracellular water were all significantly decreased (P<0.001), and visceral fat area was increased significantly after chemotherapy (P<0.05), especially in newly-diagnosed acute lymphoblastic leukemia patients after the induction of chemotherapy. CONCLUSION The nutritional status of patients with acute leukemia has undergone significant changes after chemotherapy. A single indicator has limited significance for nutritional status assessment. Comprehensive assessment of nutritional status by multiple tools is worthy of clinical application.
Acute Disease ; Humans ; Induction Chemotherapy/methods* ; Leukemia, Myeloid, Acute/drug therapy* ; Nutrition Assessment ; Nutritional Status ; Precursor Cell Lymphoblastic Leukemia-Lymphoma/drug therapy*

OBJECTIVE: To analysis the specific protein markers of essential thrombocythemia (ET) based on proteomics technology, to explore and verify the differential protein related to platelet activation. METHODS: Blood samples were obtained from ET patients and healthy people and a certain protein mass spectrometry was detected using label-free quantitative technology. The proteins relative abundance increased or down-regulated by 1.3 times in the disease group compared with the control group, and the protein abundance in the two groups t test P＜0.05 were defined as differential proteins. Bioinformatics analysis of the differential proteins was performed using GO and KEGG. The difference in the average protein abundance between the two groups was analyzed by t test and P＜0.05 was considered statistically significant. Differential proteins were selected for verification by parallel reaction monitoring (PRM) technology. RESULTS: A total of 140 differential proteins were found, of which 72 were up-regulated and 68 were down-regulated. KEGG enrichment showed that the differential protein expression was related to the platelet activation pathway. The differential proteins related to platelet activation were GPV, COL1A2, GP1bα, COL1A1 and GPVI. Among them, the expressions of GPV, GP1bα and GPVI were up-regulated, and the expressions of COL1A2 and COL1A1 were down-regulated. PRM verification of COL1A1, GP1bα, GPVI and GPV was consistent with LFP proteomics testing. CONCLUSION Differential proteins in ET patients are related to platelet activation pathway activation.Differential proteins such as GPV, GPVI, COL1A1 and GP1bα can be used as new targets related to ET platelet activation.
Blood Platelets/metabolism* ; Humans ; Platelet Activation ; Platelet Membrane Glycoproteins/metabolism* ; Technology ; Thrombocythemia, Essential