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Biomedical subjects

Yi Li

Publications and source records attributed to Yi Li.

At least 253 records · Page 14Linked to original sources

Testing the correlation for clustered categorical and censored discrete time-to-event data when covariates are measured without/with errors.

In the analysis of clustered categorical data, it is of common interest to test for the correlation within clusters, and the heterogeneity across different clusters. We address this problem by proposing a class of score tests for the null hypothesis that the variance components are zero in random effects models, for clustered nominal and ordinal categorical responses. We extend the results to accommodate clustered censored discrete time-to-event data. We next consider such tests in the situation where covariates are measured with errors. We propose using the SIMEX method to construct the score tests for the null hypothesis that the variance components are zero. Key advantages of the proposed score tests are that they can be easily implemented by fitting standard polytomous regression models and discrete failure time models, and that they are robust in the sense that no assumptions need to be made regarding the distributions of the random effects and the unobserved covariates. The asymptotic properties of the proposed tests are studied. We illustrate these tests by analyzing two data sets and evaluate their performance with simulations.

Biometry↗

[Clinical and pathological features of borderline ovarian tumors].

OBJECTIVE: To study the relationship between clinical pathological factors and prognosis. METHODS: Thirty-four cases with borderline ovarian tumors (BOT) and 30 cases with stage I ovarian epithelial cancer admitted in our hospital from Jan.1973 to Dec.2000 were studied retrospectively. All the cases were diagnosed according to the International Histological Classification and Staging of Ovarian Tumors (WHO, 1999). RESULTS: Thirty-seven cases were finally diagnosed of BOT, 6 cases with "microinvasive" had been misinterpreted as stage I ovarian cancer and one had "non-invasive peritoneal implants". Serous (38%) and mucinous (51%) tumors were dominant type of BOT and 95% of tumors were at stage I (International Federation of Gynecology and Obstetrics, FIGO). All patients were operated, 11 cases given conservative surgery with a recurrence rate of 9%, 26 cases had adjuvant chemotherapy, cyclophosphamide + adriamycin + cis-platinum used mostly. 5-year and 10 year survival rates were both 100%. The prognosis of BOT is related to pathology and adjuvant therapy. CONCLUSION: Surgery is the main treatment choice of BOT, and the criteria of chemotherapy must be adequately used.

Adolescent↗

Serum procalcitonin and interleukin-6 levels may help to differentiate systemic inflammatory response of infectious and non-infectious origin.

OBJECTIVE: To evaluate the efficacy of using procalcitonin (PCT) and interleukin-6 (IL-6) to differentiate sepsis from non-infectious systemic inflammatory response syndrome (SIRS). METHODS: We made a prospective study in a general intensive care unit at Peking Union Medical College Hospital. Twenty patients with sepsis and 31 patients with non-infectious SIRS were enrolled in this study. Serum concentrations of PCT, IL-6 and C-reactive protein (CRP) were determined within 24 h after clinical onset of sepsis or non-infectious SIRS. Leukocyte count, percentage of neutrophils, and absolute neutrophil count, as well as maximal body temperature were also recorded. RESULTS: Serum concentrations of PCT, IL-6, and CRP, as well as maximal body temperature, were significantly higher in septic patients [3.6 (1.8, 27.5) micro g/L, 810 +/- 516 ng/L, 180 +/- 108 g/L, 38.6 +/- 1.2 degrees C] than non-infectious SIRS patients [0.5 (0.2, 1.8) micro g/L, 235 +/- 177 ng/L, 109 +/- 70 g/L, 37.9 +/- 0.9 degrees C]. IL-6 and PCT exhibited the best discriminative power between sepsis and non-infectious SIRS, with sensitivity above 80% and specificity above 70%. A sepsis score with combination of IL-6 and PCT showed the best discriminative power with the area under the receiver operating characteristic curve of 0.923. CONCLUSIONS: Assessing IL-6 and PCT levels are more reliable ways to differentiate sepsis from non-infectious SIRS, compared with conventional inflammatory parameters.

Adult↗

[The fluorescence emission spectrum of NO in dielectric barrier discharge (DBD) plasma].

Dielectric barrier discharge was used to study the NO molecules of the atmosphere pollution, and the fluorescence emission spectrum in the range of 210-280 nm at low pressure was obtained in this plasma. The spectrum shows obviously the structure of double apices, whose intensity distribution in accord with Frank-Condon principle and the maximum lies at 236 nm. The main lines are attributed to the A 2 sigma+ -->X 2 pi 1/2, 2/3 transition. The produce of fluorescence can be described as: NO molecules at the ground state are impacted to the A 2 sigma+ excited state by fast electrons, and then transit to the ground state radiating fluorescence. By measuring the relation between discharge time and emission intensity of NO and N2 molecules in the discharge plasma, we demonstrated the dissociation mechanism of NO in the discharge period, which can be interpreted as: e* + NO-->N + O + e, N + NO-->N2 + O, O + NO-->NO2 + h nu.

Air Pollutants↗

[Immuno-therapeutic study of anti-idiotype minibody (single chain Fv-CH3) on ovarian carcinoma bearing mice].

OBJECTIVE: An anti-idiotypic minibody with optimal antigenicity which mimicking ovarian cancer antigen was used for therapeutic research in mice model bearing ovarian cancer. METHODS: Using gene engineering technique, prokaryotic expression vector was constructed by genetic fusion of 6B11scFv to human IgG1 hinge and CH3 region. The fusion protein named minibody was induced with IPTG in E. coli and analyzed with Western blot and inhibition ELISA tests respectively. Twenty human-PBL-SCID mice bearing i.p. Skov3.ip1 cells were divided into two groups (10 per-group), 10 mice were immunized repeatedly by minibody every two weeks for three times. Indirect ELISA test was employed for analyzing the characterization of anti-anti-idiotypic scFv (Ab3). The latent period of ascites growth and the mean survival time were observed respectively. CD4+ and CD8+ T cells from the spleen of immunized mice were assayed by flow cytometry. RESULTS: SDS-PAGE gel electrophoresis showed that a protein band with molecular weight of 50,000 appeared as the expected size after transformation and induction the host bacteria BL21 (DE3). The expressed minibody could be reacted with COC166-9 (Ab1 of 6B11) and binding goat anti-human IgG1 antibody in Western blot. Inhibition ELISA showed minibody had the capacity of binding ovarian cancer monoclonal antibody COC166-9 instead of primal antigen. Ab3 could be detected in the sera of immunized mice with minibody by ELISA test. Ab3 reached the highest at the 14th day after last vaccination and lasted for 6 weeks. The ratio of CD4+/CD8+ was the highest at the 13th day after last vaccination. The latent period of ascites growth were (37.7 +/- 5.5) days and (48.6 +/- 14.3) days (P = 0.04) respectively; while the mean survival time were (42.5 +/- 1.8) days and (59.4 +/- 16.8) days (P = 0.011) in the control and minibody group respectively. CONCLUSIONS: These results demonstrate the successful construction and expression minibody with good immune activities of 6B11scFv and human IgG1 molecules function. Antigenicity is increased without adjuvants and partial humanization is realized. Minibody can induce humoral anti-idiotypic immunity responses against ovarian carcinoma in vivo. When ascites formation was delayed or prevented and the survival was prolonged in minibody group. We expect that minibody may be used as tumor vaccine to ovarian carcinoma in the future clinical trails.

Animals↗

[Development of tumor vaccine on gynecologic cancer].

The study of tumor vaccine is one of the focus of immunological therapy on malignant gynecologic cancer. All of the ovarian carcinoma vaccine are therapeutic, including cloned antigen vaccine, tumor cell vaccine, genetic engineering tumor cell vaccine, dendritic cell (DC) vaccine, as well as anti-idiotypic vaccine. The therapeutic vaccines based on human papillomavirus (HPV) of cervical cancer are mostly summarized, including polypeptides vaccine, carrier vaccine, fusion protein or chimeric vaccine, and DC vaccine. The preventive vaccine based on HPV of cervical cancer are briefly introduced. As there are only a few reports on endometrial carcinoma vaccine.

Cancer Vaccines↗

[A study on the levels of soluble intercellular adhesion molecule-1 in the gingival crevicular fluid of the malocclusion patients with fixed appliances].

OBJECTIVE: To investigate the levels of soluble intercellular adhesion molecule-1(sICAM-1) shed into the gingival crevicular fluid (GCF) of 31 malocclusion cases, who received fixed appliances orthodontic treatment in period of pre-treatment, 1,4,8 weeks of treatment. To determine if the fixed appliance will affect the periodontal health. METHODS: The soluble intercellular adhesion molecule-1 in gingival crevicular fluid carefully collected with standardized filter strips from 31 patients with fixed appliance were measured,using an ELISA technique. The relation between the level of GCF and sICAM-1 during treatment was analyzed. RESULTS: After one week of treatment, the levels of sICAM-1 in GCF increased significantly. CONCLUSION: Using fixed appliances will affect the periodontal health. The levels of soluble intercellular adhesion molecule-1 in GCF can be used as an efficient method for monitoring periodontal health during orthodontic treatment.

Adolescent↗

[Establishment of a sandwich ELISA for detecting serum sTRAIL and its application].

AIM: To establish quantitative ELISA for detecting souble TRAIL and to evaluate its clinical application. METHODS: The mAb FMU1.1 against TRAIL was used as coating antibody, and the rabbit polyclonal antibody to TRAIL as a sandwich antibody. The serum sTRAIL levels of nine patients with hemorrhagic fever with renal syndrome and forty psoriasis patients were measured by this method. RESUITS: Sandwich ELISA was established by combination of mAb and pAb. The sensitivity of the ELISA was 0.08 microg/L. The increased levels of serum sTRAIL were found in three cases out of nine HFRS patients and eight cases out of forty psoriasis patients. CONCLUSION: A sandwich ELISA is developed, which provide an useful tool judging the patients condition, curable effect and prognosis of some related diseases.

Animals↗

[cDNA cloning and expression in Escherichia coli of rice black-streaked dwarf virus segment 7].

Using primers designed from the terminal sequences of maize rough dwarf virus S6, a 2.2 kb cDNA fragment was amplified by RT-PCR from maize plants showing maize rough dwarf disease. Sequence analysis shows that the full length of this cDNA is 2193bp. It contains two open reading frames that encoded two polypeptides with molecular weight of 41.0kD and 36.3kD, respectively. Results of multi-sequences alignment suggest that, this cDNA sequence has significant similarity to rice black-streaked dwarf virus S7, much higher than to MRDV S6. The ORFs were cloned into expression vectors, pET21-d (ORF1) or pGEX-KG (ORF2), respectively, and then transformed to BL21(DE3)-gold. After induction with IPTG, both proteins were highly expressed. The recombinant proteins were purified and high titer antisera of these two proteins were prepared.

Cloning, Molecular↗

[Cloning and expression of S9-1 gene of rice black-streaked dwarf virus in Escherichia coli].

The cDNA of RBSDV S9 was amplified by RT-PCR using the total RNA of infected maize leaves as the template, and then the PCR product was cloned into pBluescriptII SK. After the DNA sequence was determined, the S9-1 gene was subcloned into E. coli expression vector pET-21d. SDS-PAGE analysis revealed that a 40 kD protein was highly expressed. N-terminal sequence analysis confirmed the 40 kD protein was S9-1 protein. Antiserum was prepared using the purified protein. Western blotting indicated that S9-1 protein could be detected in the infected maize leaves.

Cloning, Molecular↗

Parallel cloning, expression, purification and crystallization of human proteins for structural genomics.

54 human genes were selected as test targets for parallel cloning, expression, purification and crystallization. Proteins from these genes were selected to have a molecular weight of between 14 and 50 kDa, not to have a high percentage of hydrophobic residues (i.e. more likely to be soluble) and to have no known crystal structures and were not known to be subunits of heterocomplexes. Four proteins containing transmembrane regions were selected for comparative tests. To date, 44 expression clones have been constructed with the Gateway cloning system (Invitrogen, The Netherlands). Of these, 35 clones were expressed as recombinant proteins in Escherichia coli strain BL21 (DE3)-pLysS, of which 12 were soluble and four have been purified to homogeneity. Crystallization conditions were screened for the purified proteins in 96-well plates under oil. After further refinement with the same device or by the hanging-drop method, crystals were grown, with needle, plate and prism shapes. A 2.12 A data set was collected for protein NCC27. The results provide insights into the high-throughput target selection, cloning, expression and crystallization of human genomic proteins.

Base Sequence↗

Evolution of substrate recognition across a multigene family of glycosyltransferases in Arabidopsis.

The complete sequence of the Arabidopsis genome enables definitive characterization of multigene families and analysis of their phylogenetic relationships. Using a consensus sequence previously defined for glycosyltransferases that use small-molecular-weight acceptors, 107 gene sequences were identified in the Arabidopsis genome and used to construct a phylogenetic tree. Screening recombinant proteins for their catalytic activities in vitro has revealed enzymes active toward physiologically important substrates, including hormones and secondary metabolites. The aim of this study has been to use the phylogenetic relationships across the entire family to explore the evolution of substrate recognition and regioselectivity of glucosylation. Hydroxycoumarins have been used as the model substrates for the analysis in which 90 sequences have been assayed and 48 sequences shown to recognize these compounds. The study has revealed activity in 6 of the 14 phylogenetic groups of the multigene family, suggesting that basic features of substrate recognition are retained across substantial evolutionary periods.

Arabidopsis↗

Human bone marrow stromal cell cultures conditioned by traumatic brain tissue extracts: growth factor production.

Treatment of traumatic brain injury (TBI) with bone marrow stromal cells (MSCs) improves functional outcome in the rat. However, the specific mechanisms by which introduced MSCs provide benefit remain to be elucidated. Currently, the ability of therapeutically transplanted MSCs to replace injured parenchymal CNS tissue appears limited at best. Tissue replacement, however, is not the only possible compensatory avenue in cell transplantation therapy. Various growth factors have been shown to mediate the repair and replacement of damaged tissue, so trophic support provided by transplanted MSCs may play a role in the treatment of damaged tissue. We therefore investigated the temporal profile of various growth factors, brain-derived neurotrophic factor (BDNF), nerve growth factor (NGF), vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), and hepatocyte growth factor (HGF), within cultures of human MSCs (hMSCs) conditioned with cerebral tissue extract from TBI. hMSCs were cultured with TBI extracts of rat brain in vitro and quantitative sandwich enzyme-linked immunosorbent assays (ELISAs) were performed. TBI-conditioned hMSCs cultures demonstrated a time-dependent increase of BDNF, NGF, VEGF, and HGF, indicating a responsive production of these growth factors by the hMSCs. The ELISA data suggest that transplanted hMSCs may provide therapeutic benefit via a responsive secretion of an array of growth factors that can foster neuroprotection and angiogenesis.

Analysis of Variance↗

[Serum procalcitonin and interleukin-6 help differentiate between severe sepsis and systemic inflammatory response syndrome of non-infectious origin].

OBJECTIVE: To evaluate the efficacy of serun procalcitonin (PCT) and interleukin (IL)-6 in differentiating between severe sepsis and systemic inflammatory response (SIRS) of non-infectious origin. METHOD: The serum PCT, IL-6, C-reactive protein (CRP), white blod cell count, percentage of neutrophils, and absolute neutrophil count were determined, and maximal body temperature was recorded among 21 patients was 3.6 (1.8, 27.5) micro g/L, significantly higher than that in SIRS patients (1.3 micro g/L +/- 1.6 micro g/L, P < 0.05). The IL-6 in sepsis patients was 810 ng/L +/- 516 ng/L, significantly higher than that in SIRS patients (235 ng/L +/- 177 ng/L, P < 0.01). However, the differences of CRP, WBC count, percentage of neutrophil, and absolute neutrophil count between the severe sepsis patients and SIRS patients were not statistically significant. PCT and IL-6 showed sensitivity equal or over 80% and specificity equal to or over 70%. The infection score based on PCT and IL-6 showed the best discriminative power to differentiate between severe sepsis and SIRS with the artea under the receiver operating characteristic curve of 0.923. CONCLUSION: In comparison with the conventional inflammatory markers, PCT and IL-6 are more reliable indicators to differentiate between sepsis and SIRS.

Aged↗

A prostate-derived cDNA that is mapped to human chromosome 19 encodes a novel protein.

The epithelial cells of prostate gland secrete various secretory products that play an important role in the growth and differentiation of prostate gland. These secretory products have also been implicated in neuroendocrine differentiation of benign prostatic hyperplasia and prostate malignancy. We have cloned a prostate-derived cDNA encoding a novel protein with a predicted molecular weight of 78 kDa (P(78)), and precisely mapped the cDNA sequence to chromosome 19. The P(78) gene has a complex genomic structure with 18 exons and 17 introns. The P(78) contains two conserved structural domains with limited similarity to domain D of synapsin I. The P(78) mRNA was expressed in various human cell lines. Western blot analysis using antibody specific for the P(78) revealed the presence of the P(78) protein in the prostate cancer cell lines with much lower level in metastatic prostate cancer cell lines compared to that in a primary prostate cancer cell line.

Amino Acid Sequence↗

Essential role of NF-kappa B-inducing kinase in T cell activation through the TCR/CD3 pathway.

NF-kappa B-inducing kinase (NIK) is involved in lymphoid organogenesis in mice through lymphotoxin-beta receptor signaling. To clarify the roles of NIK in T cell activation through TCR/CD3 and costimulation pathways, we have studied the function of T cells from aly mice, a strain with mutant NIK. NIK mutant T cells showed impaired proliferation and IL-2 production in response to anti-CD3 stimulation, and these effects were caused by impaired NF-kappa B activity in both mature and immature T cells; the impaired NF-kappa B activity in mature T cells was also associated with the failure of maintenance of activated NF-kappa B. In contrast, responses to costimulatory signals were largely retained in aly mice, suggesting that NIK is not uniquely coupled to the costimulatory pathways. When NIK mutant T cells were stimulated in the presence of a protein kinase C (PKC) inhibitor, proliferative responses were abrogated more severely than in control mice, suggesting that both NIK and PKC control T cell activation in a cooperative manner. We also demonstrated that NIK and PKC are involved in distinct NF-kappa B activation pathways downstream of TCR/CD3. These results suggest critical roles for NIK in setting the threshold for T cell activation, and partly account for the immunodeficiency in aly mice.

Animals↗

Association of Helicobacter pylori vacuolating toxin (VacA) with lipid rafts.

A variety of extracellular ligands and pathogens interact with raft domains in the plasma membrane of eukaryotic cells. In this study, we examined the role of lipid rafts and raft-associated glycosylphosphatidylinositol (GPI)-anchored proteins in the process by which Helicobacter pylori vacuolating toxin (VacA) intoxicates cells. We first investigated whether GPI-anchored proteins are required for VacA toxicity by analyzing wild-type Chinese hamster ovary (CHO) cells and CHO-LA1 mutant cells that are defective in production of GPI-anchored proteins. Whereas wild-type and mutant cells differed markedly in susceptibility to aerolysin (a bacterial toxin that binds to GPI-anchored proteins), they were equally susceptible to VacA. We next determined whether VacA physically associates with lipid rafts. CHO or HeLa cells were incubated with VacA, and Triton-insoluble membranes then were separated by sucrose density gradient centrifugation. Immunoblot analysis revealed that a substantial proportion of cell-associated toxin was associated with detergent-resistant membranes (DRMs). DRM association required acid activation of the purified toxin prior to contact with cells, and acid activation also was required for VacA cytotoxicity. Treatment of cells with methyl-beta-cyclodextrin (a cholesterol-depleting agent) did not inhibit VacA-induced depolarization of the plasma membrane, but interfered with the internalization or intracellular localization of VacA and inhibited the capacity of the toxin to induce cell vacuolation. Treatment of cells with nystatin also inhibited VacA-induced cell vacuolation. These data indicate that VacA associates with lipid raft microdomains in the absence of GPI-anchored proteins and suggest that association of the toxin with lipid rafts is important for VacA cytotoxicity.

Animals↗