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1.
Article in Chinese | WPRIM | ID: wpr-793277

ABSTRACT

Objective To explore the association between mild cognitive impairment (MCI) and the risk of falls in community populations, so as to provide a reference for preventing falls in the elderly. Methods 826 community residents aged 55 years and older were enrolled in Wuzhong, Ningxia; cognitive function was assessed during the participants attending the basic public health examination, and self-report falling events were observed through telephone call survey after half a year. The incidence of falls among different populations was compared using unconditional Logistic regression model. Results A total of 521 participants completed the follow-up study, and 127 of them met criteria of MCI at baseline. 32 participants reported falling occurred during follow-up with a incidence of 6.1% (32/521), and the incidence of falls was higher (8.85%,23/260) in females than that of males (3.45%,9/261) (RR=2.56, P=0.010). The incidence among MCI group (9.45%,12/127) was 1.86 times of that among non-MCI group (RR=1.86, P=0.073). Under the controlling of demographic variables (gender, age and education level), there was no association between MCI and falls (RR=1.41, P=0.382). Conclusions The incidence of falls in the elderly communities is common, and females has higher risk of falls than males. There is no statistical association between MCI and risk of falls. Further validation is needed with large-sample studies in the future.

2.
Article in English | WPRIM | ID: wpr-250339

ABSTRACT

Pancreaticoduodenectomy (PD) is the most effective treatment for patients with pancreatic head or periampullary lesions. Two major strategies exist: pylorus-preserving pancreaticoduodenectomy (PPPD) and pylorus-resecting pancreaticoduodenectomy (PRPD). However, it is yet unclear regarding the morbidity after PPPD and PRPD. This study analyzed the morbidity after PPPD and PRPD to determine the optimal surgical treatment of masses in the pancreatic head or periampullary region. A systematic search of databases identifying randomized controlled trials (RCTs) from the Cochrane Library, PubMed, EMBASE and Web of Science was performed. Outcome was compared by postoperative morbidity including overall morbidity, pancreatic fistulas, wound infections, postoperative bleeding, biliary leakage, ascites and delayed gastric emptying (DGE) rate between PPPD and PRPD. The DGE rate in the PRPD subgroups (conventional PD [CPD] and subtotal stomach-preserving PD [SSPPD], respectively) was also analyzed. The results showed that 9 RCTs including 722 participants were included for meta-analysis. Among these RCTs, 7 manuscripts described PRPD as CPD, and 2 manuscripts described PRPD as SSPPD. There were no significant differences in the overall morbidity, pancreatic fistulas, wound infections, postoperative bleeding, or biliary leakage between PPPD and PRPD. There was a lower rate of DGE with PRPD than that with PPPD (RR=2.15, P=0.03, 95% CI, 1.09-4.23). Further subgroup analysis indicated a comparable DGE rate for the CPD but a lower DGE rate for the SSPPD group than the PPPD group. However, the result did not indicate any difference between CPD and SSPPD regarding the DGE rate (P=0.92). It is suggested that PPPD is comparable to PRPD in overall morbidity, pancreatic fistulas, wound infections, postoperative bleeding and biliary leakage. The current data are not sufficient to draw a conclusion regarding which surgical procedure is associated with a lower postoperative DGE rate. Our conclusions were limited by the available data. Further evaluations of RCTs are needed.


Subject(s)
Adult , Aged , Aged, 80 and over , Humans , Middle Aged , Morbidity , Pancreaticoduodenectomy , Methods , Pylorus , General Surgery
3.
Chinese Journal of Oncology ; (12): 643-648, 2011.
Article in Chinese | WPRIM | ID: wpr-320114

ABSTRACT

<p><b>OBJECTIVE</b>To observe the expression of Toll-like receptor 8 (TLR8) in human cervical cancer cell-line HeLa cells, and the effects of TLR8 agonist CL075 on the survival and proliferation of HeLa cells.</p><p><b>METHODS</b>PCR and RT-PCR were used to detect the expression of TLR8 in 13 cancer cell lines, and the expression of COX-2, Bcl-2, VEGF mRNA in the HeLa cells stimulated by TLR8 agonist CL075 were also measured by RT-PCR. Immunofluorescence technique was used to determine the exact location of TLR8 in the cells. The percentage of viable cells was determined by trypan blue exclusion after the HeLa cells were stimulated with TLR8 agonist CL075 (0.1 µg/ml, 0.5 µg/ml, 1.0 µg/ml, 2.5 µg/ml), and cell cycle and apoptosis were analyzed by flow cytometry, and the proliferation was measured by MTT.</p><p><b>RESULTS</b>Compared with the other cancer cell lines, the expression of TLR8 in HeLa cells was the highest (703.7 ± 20.6). After stimulation by CL075, the cells had a remarkable increase of the percentage of cells in G(2)/M + S phases. In the control group, the percentage of cells in G(2)/M +S phases was (39.02 ± 2.33)%, whereas after stimulated with 1.0 µg/ml CL075, the percentage of cells in G(2)/M + S phases reached the highest ratio (57.67 ± 1.73)%, and the percentage of cells in G(2)/M + S phases had a less decrease after 2.5 µg/ml CL075 stimulation and the percentage was (56.14 ± 3.73)%. After the CL075 treatment, there was no significant changes of apoptosis compared with that of the control cells (P > 0.05), but after DDP treatment the apoptosis had a significant change (P < 0.01). After stimulation by 1.0 µg/ml CL075 for 24 h, no significant difference (P > 0.05) was found by MTT test, but a significant difference was found at 48 h and 72 h (P < 0.01). An increased expression of COX-2, Bcl-2 and VEGF mRNA was observed in HeLa cells after stimulation by TLR8 agonist CL075 for 24 h and 48 h (P < 0.05).</p><p><b>CONCLUSIONS</b>Expression of TLR8 is significantly increased in HeLa cells. The proportion of cells at different phases has a significant change after CL075 stimulation, which may up-regulate the proliferation of HeLa cells. These data suggested that TLR8 agonist may influence the tumor development and TLR8 may become a potential target in the treatment for cervical cancer.</p>


Subject(s)
Antineoplastic Agents , Pharmacology , Apoptosis , Cell Cycle , Cell Proliferation , Cisplatin , Pharmacology , Cyclooxygenase 2 , Genetics , Metabolism , Gene Expression Regulation, Neoplastic , HeLa Cells , Humans , Proto-Oncogene Proteins c-bcl-2 , Genetics , Metabolism , Quinolines , Pharmacology , RNA, Messenger , Metabolism , Thiazoles , Pharmacology , Toll-Like Receptor 8 , Genetics , Metabolism , Up-Regulation , Vascular Endothelial Growth Factor A , Genetics , Metabolism
4.
Chinese Journal of Oncology ; (12): 185-189, 2010.
Article in Chinese | WPRIM | ID: wpr-260440

ABSTRACT

<p><b>OBJECTIVE</b>To detect the expression levels of transcription factors and associated cytokines of Th17 and Treg cells in peripheral blood mononuclear cells (PBMC) of patients with gastric cancer, and explore the possible pathological mechanism of these cells involved in the development of gastric cancer.</p><p><b>METHODS</b>The mRNA levels of RORgammat, FoxP3 in PBMC were determined by quantitative real-time PCR (QRT-PCR) from 57 patients with gastric cancer, 31 patients with benign gastric illness and 40 healthy people. The concentration of IL-17, IL-23, TGF-beta, IL-10 in plasma were detected by enzyme linked immunosorbent assay (ELISA).</p><p><b>RESULTS</b>Compared with healthy volunteers, patients with gastric cancer showed higher levels of RORgammat and FoxP3 in PBMC (P < 0.05). The ratio of FoxP3/RORgammat in gastric cancer group was higher than that in the volunteer group and benign gastric illness group (P < 0.05). The ratio of FoxP3/RORgammat was higher in advanced disease than early disease (P < 0.05). The expressions of IL-17, IL-23, TGF-beta and IL-10 were higher in patients with gastric cancer than that in healthy volunteers (P < 0.05). In addition, The expression of TGF-beta and IL-10 were significantly increased in the advanced disease group than that in the early group (P < 0.05), but IL-17 and IL-23 was not significantly changed between the two groups (P > 0.05).</p><p><b>CONCLUSION</b>There are higher levels of Th17 and Treg cells in gastric cancer patients, and it also shows a persistent predominant tendency of Treg cells and a reduced tendency of Th17 cells in advanced disease. Detecting the expression of Th17/Treg transcription factor and related cytokines would contribute to the diagnosis and prediction of the disease development and prognosis.</p>


Subject(s)
Adult , Aged , Female , Forkhead Transcription Factors , Genetics , Metabolism , Gastritis , Blood , Metabolism , Pathology , Humans , Interleukin-10 , Blood , Interleukin-17 , Blood , Interleukin-23 , Blood , Male , Middle Aged , Neoplasm Staging , Nuclear Receptor Subfamily 1, Group F, Member 3 , Genetics , Metabolism , RNA, Messenger , Metabolism , Stomach Neoplasms , Blood , Metabolism , Pathology , T-Lymphocytes, Regulatory , Metabolism , Th17 Cells , Metabolism , Transforming Growth Factor beta , Blood
5.
Article in Chinese | WPRIM | ID: wpr-336194

ABSTRACT

<p><b>OBJECTIVE</b>To develop a new method of PET and CT cross-modality medical image fusion based on out-location frame.</p><p><b>METHODS</b>PET/CT cross-modality medical images were obtained based on the out-location frame and the external fiducial marker on the frame was used for rigid medical image registration. A variation model based on the wavelet transform was used for image fusion.</p><p><b>RESULTS</b>The CT images were displayed by grey scale and overlaid with the PET images displayed by chromatic scale to obtain the image after registration and fusion.</p><p><b>CONCLUSION</b>The method of external markers registration can be effective and accurate in achieving PET and CT image fusion.</p>


Subject(s)
Humans , Image Enhancement , Methods , Image Interpretation, Computer-Assisted , Methods , Positron-Emission Tomography , Methods , Radiotherapy Dosage , Radiotherapy Planning, Computer-Assisted , Methods , Radiotherapy, Computer-Assisted , Methods , Radiotherapy, Conformal , Methods , Tomography, X-Ray Computed , Methods
6.
Chinese Journal of Biotechnology ; (12): 262-268, 2004.
Article in Chinese | WPRIM | ID: wpr-259113

ABSTRACT

An E. coli expressed recombinant antigen NE2 was reported to aggregate into homo-oligomer, and can induce protective antibodies on rhesus monkey, but its immunogenicty was much weak after being purified. In this study, three N-terminal extension mutant of NE2 were expressed in E. coli, one of which named HEV 239 was found to aggregate into particle. HEV 239 antigen had good reactivity with sera of hepatitis E patients. The reactivity of HEV 239 against neutralization monoclonal antibody 8C11 was similar as NE2 antigen, while the reactivity of it against another neutralization monoclonal antibody 8H3 is much better than NE2 antigen, which indicated better antigenicity of HEV 239 than NE2. The diameter of purified HEV 239 particulate antigen was between 15 nm to 30 nm. The ED50 of immunization of HEV 239 particle adsorbed by aluminum adjuvant to BALB/c mice was between 0.08 microg to 0.25 microg. In contrast, the seraconversion rate of mice immunized by NE2 antigen adsorbed by aluminium adjuvant was only 25% on 60 microg vaccination. These results suggested that HEV 239 antigen particle has better immunogenicity as well as antigenicity than those of NE2 antigen, so it is a better vaccine candidate against HEV.


Subject(s)
Animals , Escherichia coli , Genetics , Metabolism , Female , Genetic Vectors , Genetics , Hepatitis Antigens , Allergy and Immunology , Hepatitis E virus , Genetics , Allergy and Immunology , Humans , Male , Mice , Mice, Inbred BALB C , Nucleocapsid Proteins , Genetics , Allergy and Immunology , Recombinant Proteins , Genetics , Allergy and Immunology , Viral Hepatitis Vaccines , Allergy and Immunology
7.
Article in Chinese | WPRIM | ID: wpr-305221

ABSTRACT

Hepatitis E is a main cause of acute viral hepatitis in developing countries where it occurs as sporadic cases and in epidemics form. The causative agent, hepatitis E virus, is transmitted primarily by the fecal-oral route. The approximately 7.5 kb positive-sense single-strand RNA genome includes three open reading frames (ORFs), one of which (ORF2) is postulated to encode the major viral capsid protein (pORF2) of 660 amino acid residues. We earlier showed that a bacterially expressed peptide, designated as NE2, located from amino acid residues 394 to 606 of ORF2, was found to aggregate into homodimer to at least hexamer. To understand the interface domains within this peptide vital for dimerization and formation of major neutralizing epitopes, NE2 protein underwent terminal-truncated and site-directed mutation. The hydrophobic region, ORF2 aa597-aa602 (AVAVLA), played a key role in oligomerization. Any amino acid residue of this region replaced with glutamic acid residue, the peptide can not refold as homodimer and/or oligomer. The immunoreactivities of these mutant peptides, blotted with anti-HEV neutralizing monoclonal antibody (8C11) and convalescent human sera, show associated to the formation of homodimer. The intermolecular contact region on homodimer was investigated by chemical cross-linking of two site-directed cysteines. When the alanine on aa597 site mutated with cysteine, two different homodimers were found in SDS-PAGE analysis. One (42kD) can be disassociated with 8mol/L urea, which is postulated to form by virtue of hydrophobic interaction, and the other (60kD) falls apart with the reductant DTT present. The exact conformation, generating the cross-linking reaction of cysteines, was further investigated by induced-oxidation on monomer and hydrophobic homodimer of A597C protein with GSH/GSSG. And the results revealed, it is the conformation of hydrophobic homodimer that induces the disulfide bond come into being, instead of the one of monomer. So the aa597 site was verified to be located on interface domain of hydrophobically interacting homodimeric complex. To evaluate the biological significance of hydrophobicity of interface domain, we searched natural variations as to the region on all available databases with NCBI blast program. All variations on these amino acid residues kept higher hydrophobicity, which suggests that the hydrophobic domain is critical for the assemblage and propagation of HEV. NE2 N-terminal deletions up to aa458 had no effect on dimerization and took no exact part in formation of major neutralizing epitopes, but the fragment may act as helper for the formation of major neutralizing epitopes on NE2. Interestingly, the C-terminus aa605-aa660 of ORF2 can also act as helper instead of the N-terminus of NE2. This study suggests an interface domain of NE2 might be vital for HEV capsomer assembly and formation of major neutralizing epitopes. These results may offer clues to the rational design of recombinant anti-HEV vaccine.


Subject(s)
Capsid Proteins , Chemistry , Hepatitis E virus , Chemistry , Hydrophobic and Hydrophilic Interactions , Protein Multimerization , Protein Structure, Tertiary , Virus Assembly
8.
Article in Chinese | WPRIM | ID: wpr-240527

ABSTRACT

<p><b>OBJECTIVE</b>To evaluate the serological markers and biological marker in the diagnosis of hepatitis E infection in a rhesus monkey model.</p><p><b>METHODS</b>86 rhesus monkeys had been infected with different doses of HEV. Hence, they were taken sequential blood samples at intervals up to 86 weeks for 4 hepatitis E virus (HEV) specific antibody assays (E2-IgM, E2-IgG, GL-IgG, and YES-IgG), and nucleic acid assay.</p><p><b>RESULTS</b>All the animals produced E2-IgG and all but one also produced E2-IgM and excreted the virus in stool, whereas positive rate of GL-IgG and YES IgG were low and correlated with virus level. Hepatitis occurred over a period of 4 weeks (between 3 an 7 weeks) after infection. Virological marker occurred mainly during incubation period and declined rapidly after onset of hepatitis. Seroconversion of E2-IgM occurred before onset of hepatitis in 70% monkeys and declined rapidly up to 50% of peak value after 4 weeks. E2-IgM seroconversion was closely paralleled by E2-IgG; however, E2-IgG persisted in all animals for the entire duration of experiment of up to 86 weeks. Production of GL-IgG and YES-IgG was delayed by one week after the E2 antibodies, these antibodies showed a transient occurrence and seroprevalence declined to 50% of the peak value over a period of 12 weeks.</p><p><b>CONCLUSION</b>E2-IgM might be used as a suitable acute hepatitis E marker, and E2-IgG as a suitable epidemiological marker. The seroconversion or titer elevation of GL-IgG and YES-IgG antibodies probably used to confirm the infection. The viral markers are optional for early diagnosis.</p>


Subject(s)
Alanine Transaminase , Blood , Animals , Biomarkers , Genotype , Hepatitis Antibodies , Blood , Hepatitis E , Diagnosis , Hepatitis E virus , Classification , Genetics , Allergy and Immunology , Immunoglobulin E , Blood , Immunoglobulin M , Blood , Macaca mulatta
9.
Chinese Journal of Biotechnology ; (12): 450-455, 2004.
Article in Chinese | WPRIM | ID: wpr-249964

ABSTRACT

Production of Hepatitis E Virus capsid protein by high cell density culture in recombinant E. coli has been studied in 10L and 30L fermentors. The effects of different factors on growth and producing recombinant protein of E. coli have been studied by batch culture, such as different media, the ratio of phosphate and Magnesium sulfate. Comparison of fermentation performance for recombinant E. coli in different fed-methods culture has been investigated by fed-batch culture. The effects of inducing at different stages of growth and time of inducing on growth and producing recombinant protein, also obtained by fed-batch culture. At last, the solubility of inclusion body in different urea concentrations also has been obtained by fed-batch culture. The results show that the concentration of phosphate and Magnesium sulfate in the optimal media is 80mmol/L and 20mmol/L in batch culture respectively, that induction with 1.0mmol/L IPTG at mid log phase (about 45 OD at 600nm) is suitable for growth and recombinant protein expression, the cells were approaching stationary growth phase and the maximum cell OD at 600nm of 80 was achieved in 5h of fed-batch culture, and the expression level is 29.74%. The results also indicate that the solubility of inclusion body in 4mol/L urea solution induced at 37 degrees C reaches 14mg/mL, over 80% inclusion body was resolved. The culture process achieved in 10L fermentor could be successfully scaled up to 30L fenmentor with good reproducibility.


Subject(s)
Bioreactors , Microbiology , Colony Count, Microbial , Escherichia coli , Genetics , Metabolism , Hepatitis E virus , Genetics , Nucleocapsid Proteins , Genetics , Protein Engineering , Methods , Recombinant Fusion Proteins , Genetics
10.
Chinese Journal of Biotechnology ; (12): 463-467, 2002.
Article in Chinese | WPRIM | ID: wpr-256184

ABSTRACT

A fragment of hepatitis E virus open reading frame-2(ORF2), located from amino acid residues 394 to 604, was expressed in E. coli. The recombinant protein NE2 was found to form homodimer mostly in SDS-PAGE, which can be dissociated to monomers when treated with urea, and it was recognized more strongly in its dimeric form than the monomer by HEV reactive human serum in Western blotting. Besides, many aggregated form of NE2 from dimer to at least hexamer can be seen in MALDI-TOF-MS. And when the hydrated dynamic semidiameter of NE2 moleculars in PBS was measured as about 4 nm by Dynamic Light Scattering (DLS), being equal to tetramer, but with high polydispersity, which suggested that the NE2 moleculars were existed in PBS in many different sizes. These results suggested that the recombinant NE2 can aggregate into several oligomer forms, the association in the dimer is most strong, and dimers can assemble further to form some super-structure.


Subject(s)
Dimerization , Electrophoresis, Polyacrylamide Gel , Escherichia coli , Genetics , Gene Expression , Protein Conformation , Recombinant Proteins , Chemistry , Genetics , Metabolism , Viral Proteins , Chemistry , Genetics , Metabolism
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