Biomedical subjects

Jing Zhao

Publications and source records attributed to Jing Zhao.

At least 163 records · Page 9Linked to original sources

PubMed2003

The antioxidant effects of ribonuclease inhibitor.

Ribonuclease inhibitor (RI) is an acidic cytosolic glycoprotein with molecular weight of about 50 kDa, which contains 32 cysteine residues. It is possibly that RI may have antioxidant effect by thiol-disulfide exchange reaction. We studied the effects of RI over-expression on the rat glial cell line C6 injured with H2O2. The transfected C6 cells with RI cDNA (C6') had higher viability, less LDH leakage and MDA contents, but more GSH contents compare that in the control C6 cells. In transfected C6 cells, the activities of CAT and GST were higher than that in the control C6 cells. Without H202 stress, the activities of CAT and GST in the C6' cells were 1.73 and 3.62 times that in the control C6 cells, respectively; With 1.00 mmol/L H2O2 stress, the activities of CATand GSTin the C6' cells were 3.38 and 2.11 times that in the C6 cells, respectively. These results suggest that the over-expression RI has antioxidant activity and it is able to protect cells from per-oxidative injuries. Moreover, we investigated whether RI has a protective role against mouse hepatic damage in vivo. The mice pretreated with different doses of human RI were injected by CC14. The results show that the SOD activities of therapy groups were significantly higher than that of the control group (p < 0.01), while the contents of MOD and activities of ALT and AST in blood were remarkably lower than that of the control group (p < 0.01). Pathological examination shows that the degree of damage was alleviated with RI therapy. These results suggest that RI has the protective role against mouse hepatic damage induced by CC14. The anti-oxidative effects of RI may play an important role in cell protection from per-oxidative injuries.

Alanine Transaminase↗

PubMed2003

Dispersive pad site burns with modern radiofrequency ablation equipment.

Pad burns during or after radiofrequency ablation (RFA) are a skin complication probably underreported in the literature. We report on 4 severe pad burns, deep second and third degree, in 3 patients undergoing radiofrequency ablation of liver malignancies, 1 percutaneously and the other 2 after laparotomy. All burns occurred at the leading edge or at the corner of the pads attached to the patients' thighs. Potential causes leading to the burns are discussed. Current dispersive pad designs do not prevent the leading edge phenomenon and subsequent burns. Further developmental work in the pad design with the possibility of skin temperature monitoring via temperature sensors under the leading pad edge is needed.

Aged↗

PubMed2003

Isolation and some characterizations of a glycosylated fibrinolytic enzyme of earthworm, Eisenia fetida.

Resin coupled with m-aminophenylbornic acid was used to isolate a glycosylated component from homogenate of earthworm (Eisenia fetida). The fraction showed a single band on SDS-PAGE with a molecular weight of 34, 193 Da determined by mass spectroscopy. The N-terminal region is AQVCCPDI, different from those of earthworm fibrinolytic enzymes reported previously (Nakajima et al. 1993). This glycosylated component showed an activity on digesting both Chromozym-TH and fibrin, suggesting that it is a novel fibrinolytic enzyme.

Animals↗

PubMed2003

Mutation analysis of novel human liver-related putative tumor suppressor gene in hepatocellular carcinoma.

AIM: To find the point mutations meaningful for inactivation of liver-related putative tumor suppressor gene (LPTS) gene, a human novel liver-related putative tumor suppressor gene and telomerase inhibitor in hepatocellular carcinoma. METHODS: The entire coding sequence of LPTS gene was examined for mutations by single strand conformation polymorphism (SSCP) assay and PCR products direct sequencing in 56 liver cancer cell lines, 7 ovarian cancer and 7 head neck tumor cell lines and 70 pairs of HCC tissues samples. The cDNA fragment coding for the most frequent mutant protein was subcloned into GST fusion expression vector. The product was expressed in E.coli and purified by glutathione-agarose column. Telomeric repeat amplification protocol (TRAP) assays were performed to study the effect of point mutation to telomerase inhibitory activity. RESULTS: SSCP gels showed the abnormal shifting bands and DNA sequencing found that there were 5 different mutations and/or polymorphisms in 12 tumor cell lines located at exon2, exon5 and exon7. The main alterations were A(778)A/G and A(880)T in exon7. The change in site of 778 could not be found in HCC tissue samples, while the mutation in position 880 was seen in 7 (10 %) cases. The mutation in the site of 880 had no effect on telomerase inhibitory activity. CONCLUSION: Alterations identified in this study are polymorphisms of LPTS gene. LPTS mutations occur in HCC but are infrequent and of little effect on the telomerase inhibitory function of the protein. Epigenetics, such as methylation, acetylation, may play the key role in inactivation of LPTS.

Carcinoma, Hepatocellular↗

PubMed2003

Effect of caffeic acid phenethyl ester on proliferation and apoptosis of hepatic stellate cells in vitro.

AIM: To investigate the role of nuclear factor-kappaB (NF-kappaB) inhibitor caffeic acid phenethy1 ester (CAPE) in the proliferation, collagen synthesis and apoptosis of hepatic stellate cells (HSCs) of rats. METHODS: The HSCs from rats were isolated and cultured in Dulbecco's Modified Eagle's Medium (DMEM) and treated with CAPE. The proliferation and collagen synthesis of HSCs were determined by (3)H-TdR and (3)H-proline incorporation respectively, and the expression of type I, III procollagen genes was further explored by in situ hybridization. Apoptosis cell indices (AIs) were examined using terminal deoxynucleotidyl transferase- mediated DIG-dUTP nick end labeling (TUNEL). RESULTS: In activated HSC in culture, CAPE significantly inhibited (3)H-TdR and (3)H-proline incorporation by HSCs at concentrations of 5 micromol/L and 10 micromol/L respectively. CAPE also reduced the type I procollagen gene expression (P<0.05) at higher concentration. Apoptosis of HSC was induced by CAPE and the AIs were time-and dose-dependently increased from 2.82+/-0.73 % to 7.66+/-1.25 % at 12 h (P<0.01) and from 3.15+/-0.88 % to 10.61+/-2.88 % at 24 h (P<0.01). CONCLUSION: CAPE inhibits proliferation and collagen synthesis of HSC at lower concentration and induces HSC apoptosis at higher concentration.

Animals↗

PubMed2003

[Prevalence of respiratory and atopic disorders in Chinese school children].

OBJECTIVE: To compare the prevalence of respiratory and atopic disorders, and to assess the role of atopy in the development of asthma in Chinese school children from Beijing, Guangzhou, and Hong Kong. METHODS: Community-based random samples of school children aged 9 approximately 11 years from three Chinese cities (Beijing, Guangzhou and Hong Kong) were recruited for study using the International Study of Asthma and Allergies in Childhood (ISAAC) Phase II protocol. Subjects were studied by parental questionnaires (n = 10 902), skin-prick tests and skin examination (n = 3 479). RESULTS: The prevalence rates of current wheeze, speech limiting wheeze, rhinoconjunctivitis and flexural dermatitis were significantly higher in Hong Kong than in Beijing and Guangzhou. The atopy rate was also higher in Hong Kong (41.2%) than in Beijing (23.9%) and Guangzhou (30.8%). Atopy was strongly correlated with current wheeze (OR = 7.74; 95% CI = 5.70 approximately 10.51). Subgroup analyses of children from Hong Kong revealed that children born in mainland China who had subsequently migrated to Hong Kong had a significantly lower rate of allergic symptoms and atopy than those born in Hong Kong. CONCLUSION: The prevalence of asthma, allergic diseases and atopy was highest in school children from Hong Kong. Atopic sensitizations is an important factor associated with asthma in Chinese children.

Animals↗

PubMed2003

[Relationships between the diameters of Bacille Calmette-Guerin scars and asthma, atopy in urban and rural Beijing children].

OBJECTIVE: To compare the diameters of Bacille Calmette-Guerin (BCG) scars between asthmatic (allergic) students and normal students, and to compare the diameters of BCG scars in children in rural Beijing area with those in urban Beijing area, and to compare the prevalence of asthmatic (allergic) symptoms and atopy between rural and urban Beijing students. METHODS: Sampled by cluster-random Method, 819 urban school children and 1 044 rural school children aged 13 - 14 years were recruited for the Beijing arm of the phase III international study of asthma and allergies in childhood. The children answered the core questionnaires, and underwent skin prick test for 13 allergens, and were checked for BCG scars on their arms. RESULTS: (1) In both rural and urban Beijing students, the mean diameters of BCG scars in those with asthma (allergy) (asthma ever, wheeze ever, current wheeze within 12 months, hay fever ever, repeated rash ever, eczema ever, allergic rhinitis, atopy, high-sensitivity or were diagnosed asthma) were (7.0 +/- 2.1) mm-(8.8 +/- 2.8) mm, which were not significantly different from those of (7.2 +/- 2.7) mm-(8.2 +/- 2.2) mm in normal students (P > 0.05). (2) Compared to the urban students, the mean diameter of BCG scars in the rural students was (8.2 +/- 2.3) mm, significantly larger than that of (7.3 +/- 2.9) mm in the urban students (P < 0.01). The prevalence of asthma (allergy) (asthma ever, wheeze ever, current wheeze within 12 months, hay fever ever, repeated rash ever, eczema ever, allergic rhinitis, atopy, high-sensitivity or diagnosed asthma) were 0.6% - 11.4% in the rural Beijing students, significantly lower than that of 3.1% - 35.7% in urban Beijing schoolchildren (P < 0.01). CONCLUSIONS: The diameters of BCG scars were not significantly different between asthmatic (allergic) students and normal students. The diameters of BCG scars in the rural students were significantly larger than those in the urban students. The prevalence of asthma and allergy in the rural children were significantly lower than that in the urban students. The results suggest that there is difference in immunity between rural and urban children from early life, which may be caused by the different living environments.

Adolescent↗

PubMed2003

[An epidemiological study on sleep problems in children aged 1 to 23 months in Shanghai].

OBJECTIVE: To understand prevalence of sleep problems and presence of improper sleep habit of children aged one to 23 months in Shanghai, and to identify their related factors. METHODS: Totally, 1 252 normal children, without severe illness and abnormal delivery history, aged one to 23 months were randomly selected from five districts of Shanghai. Under strict quality control system, their parents were interviewed with questionnaire to look into their familial and social environment, as well as sleep-related factors. RESULTS: Fully-completed information was obtained in 1 129 of 1 252 children, with a response rate of 90.18%. Prevalence of sleep problems in children was 65.90% in Shanghai, remarkably higher than in the other areas. Prevalence of nightmare, level of fright at night, snore and sleep apnea in boys were significantly higher than those in girls. Prevalence of improper sleep habit in children was also higher, including sleeping with their parents, parental presence at sleep onset and irregular sleep time. Main risk factors related to children's sleep problems were maternal anxiety, children's irregular sleep time, parental presence at sleep onset, difficulty-type temperament and maternal poor sleep during pregnancy. CONCLUSIONS: In Shanghai, prevalence of sleep problems in children aged one to 23 months was relatively higher and main risk factors for them were social, cultural and familial environment, especially negligence of cultivating proper sleep habit and parental improper response to children's poor sleep behavior.

Adaptation, Psychological↗

PubMed2003

[Individual allergens as risk factors for asthma and bronchial hyperresponsiveness in Chinese children].

OBJECTIVE This study aimed to determine the relationship between individual allergens with current wheezing and bronchial hyperresponsiveness (BHR) in schoolchildren from three chinese cities: Beijing, Guangzhou and Hong Kong. METHODS Community-based random samples of 10-yr-old schoolchildren from the 3 cities were recruited for study using the International Study of Asthma and Allergies in Childhood (ISAAC) Phase II protocol. The subjects were studied by parental questionnaires (n = 10,902), skin-prick tests (n = 3478), and methacholine challenge tests (n = 608). RESULTS The highest prevalence rates of wheezing in the past 12 months (Beijing, 3.8%; Guangzhou, 3.4%; Hong Kong, 5.8%) and atopy (Beijing, 23.9%; Guangzhou, 30.8%; Hong Kong, 41.2%, defined as having <or= 1 positive skin-prick tests,) were seen in schoolchildren from Hong Kong. The highest prevalence rates of BHR (Beijing, 33.2%; Guangzhou, 45.8%; Hong Kong, 30.7%) were seen in schoolchildren from Guangzhou. Multivariate-logistic regression analyses revealed that sensitization to Dermatophagoides pteronyssinus [odds ratio ((OR)) = 4.48; 95% confidence interval ((CI)); 3.02-6.66], cat ((OR) 2.85; 95% (CI) 1.24-6.50), Dermatophagoides farinae (DF) ((OR) 2.41, 95% (CI) 1.65-3.51) and Mixed grass pollen ((OR) 2.85, 95% (CI) 1.24-6.50) were significantly associate with current wheezing atopy ((OR) 1.29, 95% (CI) 0.74-2.24), was not an independent risk factor for current wheezing in children from any of the three cities. Furthermore, atopy ((OR) 2.53, 95% (CI) 1.93-6.97) were significantly associated with BHR. CONCLUSION The authors confirmed that sensitization to house dust mite and cat was significantly associated with current wheezing and bronchial hyperresponsiveness in Chinese schoolchildren.

Allergens↗

PubMed2003

[Effect of the truncated human apoptosis-inducing factor expression on apoptosis of HeLa cells].

AIM: To observe the expression of the truncated human apoptosis-inducing factor (AIF) gene and its apoptosis-inducing effect on HeLa cells. METHODS: Full-length human AIF gene was cloned by RT-PCR, then the truncated AIF gene was constructed by deleting the N-terminal mitochondrial location sequence (MLS), and inserted into the EGFP co-expression vector pIRES2-EGFP. After being transfected into HeLa cells with Lipofectamin, the expression of the truncated AIF gene and its effect on HeLa cells morphology and growth condition were detected by fluorescence microscope, immunohistochemical staining, indirect fluoroimmunoassay and electron microscope analysis. RESULTS: The eukaryotic expression vector pIRES2-EGFP containing of truncated human AIF gene was constructed successfully. The AIF protein could be detected in the transfected HeLa cells. After transfection, typical apoptotic feature of the transfected HeLa cells and a mass of cell death were observed under electron microscope. CONCLUSION: The expression of the truncated human AIF gene can induce apoptosis of the transfected human HeLa cells.

Apoptosis↗

PubMed2003

[Expression of a secreted fusion protein consisting of anti-HER2 antibody and reversed caspase-3 and its suppression on growth of SKOV3 cells].

AIM: To investigate the killing effect on SKOV3 cells by fusion protein HER2-specific antibody-reversed caspase-3 in the secreted form. METHODS: The reversed human caspase-3 gene was subcloned into pCMV-e23scFv-PEII-PEIII to construct recombinant eukaryotic expression vector pCMV-e23scFv-PEII-revcasp-3 and transfect Jurkat cells. The secreted expression of the in culture supernatant of transfected Jurkat cells was detected by ELISA. The suppression of the fusion protein on growth of SKOV3 cells was examined by co-culture with supernatant of transfected Jurkat cells. RESULTS: There was the secreted expression of the fusion protein in Jurkat cells. Expressed product had the activity of inducing SKOV3 cells to death. CONCLUSION: The fusion protein expressed in the secreted form can targeted towards SKOV3 cells and kill them.

Antibodies↗

PubMed2003

[Construction of single-chain Fv Gene of Anti-HBsAg and its expression in COS-7 cells].

AIM: To subclone the V(L) gene and V(H) genes of anti-HBsAg and construct the single-chain Fv gene and to analyse the expression of the constructed gene in COS-7 cells. METHODS: A set of oligonucleotide primers were designed and used to amplify the V(H) and V(L) genes from Fab antibodies screened from phage antibody library. The products were cloned into pUC19 vector and their sequences were analysed. The V(H) and V(L) gene fragments were linked up by a peptide linker and a leader sequence added at 5' terminus of each gene (L-V(H)-Linker-V(L)) and designated as L-ScFv. The L-ScFv genes were inserted into the eukaryotic fusion protein expression vector pEGFP-N3 and transfected into COS-7 cells to express respectively. RESULTS: The coding sequences of V(H),V(L), linker and leader in all constructs were all correct. The expression of ScFv fusion protein was detectable by fluorescence microscope after transient expression in COS-7. The secretive expression of L-ScFv was confirmed by SDS-PAGE and Western blot analysis. And the binding specificity of the ScFvs with HBsAg were proved by indirect ELISA. CONCLUSIONS: Anti-HBsAg ScFv genes have been successfully constructed and expressed in COS-7 cells.

Animals↗

PubMed2003

[The significance of fluorescent patterns of antineutrophil cytoplasmic antibodies (ANCA) in renal diseases].

AIM: To further explore significance of fluorescent patterns of antineutrophil cytoplasmic antibodies in renal diseases. METHODS: ANCA fluorescent patterns and its target antigens in sera of 251 patients with renal diseases were detected by indirect immunofluorescence (IIF) and ELISA. RESULTS: The positive percentage of ANCA is 20.3% (51/251), in which 3.98%(10/251) is cANCA, 11.95% (30/251) pANCA and 4.38%(11/251) aANCA. CONCLUSION: The simultaneous screening for ANCA and its target antigens by IIF and ELISA can increase the positive percentage of ANCA in renal diseases. This novel screening strategy is able to distinguish atypical ANCA(aANCA) from pANCA with ANA and therefore useful for differential diagnosis between vasculitis and autoimmune diseases.

Antibodies, Antineutrophil Cytoplasmic↗

PubMed2003

[Construction and expression of recombinant antibody/granzyme B containing truncated translocating peptide].

AIM: To construct an eukaryotic expression vector for recombinant antibody/granzyme B gene containing truncated translocating peptide and express it in HER-2(+) SKBR-3 cells and HER-2(-) Hela cells. METHODS: PCR amplication was used to obtain recombinant DNA encoding antibody/granzyme B containing truncated translocating peptide DNA, and then the DNA fragment was cloned into eukaryotic expression vector pCMV-e23sFv-PE40. After being transfected into SKBR-3 cells and Hela cells, the expression of target gene and its effect on cellular morphology were detected by immunocytochemical staining. RESULTS: The eukaryotic expression vector encoding recombinant antibody/granzyme B containing truncated translocating peptide was successfully constructed. e23sFv-FSD-GrB protein was expressed in most of Hela cells and had no effect on Hela cells, but the transfected SKBR-3 cells which also express e23sFv-FSD-GrB protein exhibited condensed nucleus and cytoplasm. CONCLUSION: The construction of eukaryotic expression vector of recombinant antibody/granzyme B gene containing truncated translocating peptide lays the foundation for further determination of minimal fragment responsible for translocation.

ADP Ribose Transferases↗

PubMed2003

[Prokaryotic expression and bioactivity identification of Fab against human gamma-seminoprotein].

AIM: To construct the expression vector containing cDNA encoding Fab against human gamma-seminoprotein and express it in E. coli. METHODS: The genes encoding K chain and Fd against gamma-seminoprotein were acquired from pUC19-K and pBluescript KS( M13-)-Fd by restrictive enzyme digestion and then cloned into the expression vector pComb3 to construct recombinant expression vector pComb3-Fab. pComb3-Fab was transfected into and expressed in XLI-Blue. RESULTS: Fab against r-semino-protein was expressed in. XLI-Blue. Western blot analysis and immunocytochemical staining demonstrated that ex-pressed Fab could specifically bind to gamma-seminoprotein. CONCLUSION: Fab against gamma -seminoprotein has been expressed successfully with biological activity, which create favourable condition for further study on targeted therapy of prostate cancer.

Animals↗

PubMed2003

[A study of correlation between the change of chromosome centromeric dots and habitual abortions].

To study the correlation between chromosome centromeric dots and habitual abortions, Cd variation, Cd loss,maximum diameter of Cd and Cd-NOR of 38 habitual abortion patients and 42 healthy persons were measured, compared and analysed with Cd-banding technique. It was found that the frequencies of Cd variation and Cd loss were obviously higher and maximum diameter of Cd was smaller in habitual abortion patients than those in healthy persons. The increase of frequencies of Cd variation and Cd loss and the decrease of maximum diameter of Cd might be the causes affecting habitual abortions.

English Abstract↗

PubMed2003

[Establishment and biological properties of hybridoma cell lines secreting anti-IBDV idiotypic antibodies].

In recent years, the prevention and cure of infectious bursal disease (IBD) have become more and more difficult due to the emergence of very virulent strains of infectious bursal disease virus (vvIBDV) and the variant strains of IBDV. In this research, the hybridoma cell lines which secretes anti-idiotypic antibodies against anti-IBDV IgG were established. According to the Jerne's theory of immune network, the use of the anti-idiotypic antibodies as a vaccine will be a new method for the prevention of IBD. In this study, the SPF chickens were inoculated with the IBDV- SD strain, and the bursal was obtained from the died chickens. The bursal was then homogenized and frozen-thawed 3 cycles, and the virus samples were prepared by cane sugar density gradient centrifugation and dialysis. Typical IBDV particles were observed under an electron microscope, and the concentration of the virus protein measured by ultraviolet absorbance spectrophotometry was 10.8 mg/mL. SPF chickens were immunized with the virus and the highly immunized sera were prepared and purified by Sulfuric acid ammonia salt out and Sephadex G-25 chromatography. Then, Balb/C mice of six or eight weeks old were immunized interapertoneally(I. P.) with purified antibodies to IBDV at regular intervals. SP2/0 myeloma cells were fused with the spleencytes from the immunized mice at a ratio of 10:1, in 50% polyethylene glycol (1540) and were then cultured in HAT until all the SP2/0 cells died. The hybridoma cells were selected by ELISA and the highly positive holes were cloned 3 times with the method of limited dilution. Two strains (2B6 strain,5F4 strain) of hybridoma cells were obtained, which were shown by ELISA to steadily secrete anti-IBDV idiotypic antibodies. The chromosome number of the two hybridoma cells were about 88 - 106, 95 in average, and the antibodies secreted belonged to the types of IgG1 and Kappa. Balb/c mice of 3 months old were inoculated I.P. with about 10(7) hybridoma cells per capita, and the ascites were collected 12 days later and the titre of anti-IBDV idiotypic antibodies measured by ELISA was 1 :25600 (for 2B6) and 1:12800 (for 5F4) . The ascites containing the anti-IBDV idiotypic antibodies were emulsified with complete or incomplete Freund's adjuvants, and the anti-IBDV idiotypic antibody vaccine was obtained. SPF and common Jingbai chickens were immunized with the vaccine obtained. The immunized chickens with the vaccine were inoculated with IBDV-SD strain at a dose of 2000 ELD50 after twoimmunizations. All the 10 SPF chickens in the non-immunized group were sick, and 8 of them died; and 5 out of the 50 SPF chickens immunized group got sick and 2 died. All the 10 common Jingbai chickens in the control group were sick, and 6 died; 7 of the 30 immunized common Jingbai chickens got sick and only 1 died. Chi2 analysis showed that the difference between the immunized and the non-immunized groups in both the SPF and the common Jingbai chickens were significant (P < 0.01). Our result indicated that the anti-IBDV idiotypic antibody vaccine well protected chickens and had a great potential in both research and clinical application.

Animals↗

PubMed2002

Effect of alpha2M on earthworm fibrinolytic enzyme III-1 from Lumbricus rubellus.

Though it is known that human alpha2-macroglobulin (alpha2M) inhibits most proteases, the effect of alpha2M has not been investigated on earthworm fibrinolytic enzymes (EFEs) from Lumbricus rubellus, which could be transported from intestine epithelium into blood as an intact molecule (Fan et al., Biochim. Biophys. Acta 1526 (2001) 286). The activity of earthworm fibrinolytic III-1 (EFE-III-1) decreased to 65% when incubated with alpha2M, while it decreased to 30% in plasma under the same conditions. The first order rate of the inactivation of EFE-III-1 with alpha2M was similar to that of fast phase with plasma, indicating that alpha2M may be the inhibitor initially binding to the enzyme in blood. SDS-PAGE showed that incubation of EFE-III-1 with alpha2M a released fragment ( approximately 90 kDa), followed by formation of a high molecular weight complex (approximately 700 kDa). There was a linear relationship between the apparent inhibition rate constant (k1) and [alpha2M], by double reciprocal plot. It was suggested, as described by Tsou (Acta Biochem. Biophys. Sinica 5 (1965) 398) and Tian (Biochem. J. 21 (1982) 1028), that the mechanism of alpha2M/EFE-III-1 interaction could be coincided with a complexing irreversible inhibition. Experiments in both the inactivation and the intrinsic fluorescence showed that alpha2M bound to the enzyme mole by mole equivalently. The intrinsic fluorescence of alpha2M was enhanced with an observable blue shift in emission maxima, suggesting that alpha2M was one of the important inhibitors to EFEs when it absorbed into blood.

Animals↗