Functional proteomic analysis of the protein kinase C signaling system.
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Animals↗
Biomedical subjects
Jun Zhang
Publications and source records attributed to Jun Zhang.
Cardiac toxic effects of trans-2-hexenal are mediated by induction of cardiomyocyte apoptotic pathways.
Aldehydes are ubiquitous pollutants with well-indicated but ill-defined cardiovascular toxicity. To investigate the direct toxic effects of environmental aldehyde exposure on the myocardium, 8-wk-old male ICR (Institute of Cancer Research) strain mice were gavage fed trans-2-hexenal (0.1, 1, 10, or 50 mg/kg/wk) or corn oil (vehicle) for 4 wk, during which cardiac function, myocardial morphology, cardiomyocyte apoptosis, and the cytochrome cmediated caspase activation apoptotic pathway were determined. Quantification by enzyme-linked immunosorbent assay (ELISA) revealed that aldehyde- protein adducts increase in mouse hearts following hexenal treatment, whereas echocardiographic analysis displayed a significant impairment of basal left-ventricular contractile function. Both histological analysis and TUNEL (terminal deoxynucleotidyl transferase-mediated nick-end labeling) staining indicated condensed nuclei and a significant increase in cardiomyocyte apoptosis in these mice, but immunohistochemistry-based confocal microscope revealed no marked myofibril disarray. Release of cytochrome c from mitochondria into the cytosol, concomitant with activation of caspase-3 and -9, was also found in hexenal-treated groups. In addition, isolated cardiac mitochondria formed hexenal-protein adducts when treated with hexenal, providing indirect evidence that the cardiac mitochondrion is one of primary subcellular targets of aldehyde toxins. These findings suggest that trans-2-hexenal exposure results in direct cardiac toxicity through, at least in part, induction of mitochondrial cytochrome c release-mediated apoptosis in cardiomyocytes, indicating that the cardiac mitochondrion is one of principal subcellular targets of aldehyde toxins.
Actinin↗
DcR3/TR6 effectively prevents islet primary nonfunction after transplantation.
Islet primary nonfunction (PNF) is defined as the loss of islet function after transplantation for reasons other than graft rejection. It is a major obstacle to successful and efficient islet transplantation. DcR3/TR6 is a soluble death decoy receptor belonging to the tumor necrosis factor (TNF) receptor family, and it can block apoptosis mediated by several TNF receptor family members such as Fas and LT beta R. In this study, we used TR6 to protect islets from PNF after transplantation. Untreated isogeneic or allogeneic islet transplantation had PNF incidence of 25 and 26.5%, respectively. Administration of TR6 totally prevented PNF in allogeneic islet transplantation. In vitro experiments showed an increased apoptosis among islets that were treated with FasL and gamma-interferon (IFN-gamma) in combination. TR6 significantly reduced such apoptosis. Functional study showed that insulin release was compromised after FasL and IFN-gamma treatment, and the compromise could be prevented with TR6-Fc. This indicates that TR6 indeed protected beta-cells from damage caused by FasL and IFN-gamma. Further in vivo experiments showed that syngeneic islet transplantation between lpr/lpr and gld/gld mice was significantly more efficacious than that conducted between wild-type mice. These results suggest that Fas-mediated apoptosis plays an important role in PNF, and use of TR6 may be a novel strategy to prevent PNF in clinical islet transplantation.
Animals↗
Expression of ORF2 partial gene of hepatitis E virus in tomatoes and immunoactivity of expression products.
AIM: To transfer hepatitis E virus (HEV) ORF2 partial gene to tomato plants, to investigate its expression in transformants and the immunoactivity of expression products, and to explore the feasibility of developing a new type of plant-derived HEV oral vaccine. METHODS: Plant binary expression vector p1301E2, carrying a fragment of HEV open reading frame-2 (named HEV-E2), was constructed by linking the fragment to a constitutive CaMV35s promoter and nos terminator, then directly introduced into Agrobacterium tumefaciens EHA105. With leaf-disc method, tomato plants medicated by EHA105 were transformed and hygromycin-resistant plantlets were obtained in selective medium containing hygromycin. The presence and integration of foreign DNA in transgenic tomato genome were confirmed by Gus gene expression, PCR amplification and Southern dot blotting. The immunoactivity of recombinant protein extracted from transformed plants was examined by enzyme-linked immunosorbant assay (ELISA) using a monoclonal antibody specifically against HEV. ELISA was also used to estimate the recombinant protein content in leaves and fruits of the transformants. RESULTS: Seven positive lines of HEV-E2-transgenic tomato plants confirmed by PCR and Southern blotting were obtained and the immunoactivity of recombinant protein could be detected in extracts of transformants. The expression levels of recombinant protein were 61.22 ng/g fresh weight in fruits and 6.37-47.9 ng/g fresh weight in leaves of the transformants. CONCLUSION: HEV-E2 gene was correctly expressed in transgenic tomatoes and the recombinant antigen derived from them has normal immunoactivity. Transgenic tomatoes may hold a good promise for producing a new type of low-cost oral vaccine for hepatitis E virus.
Administration, Oral↗
Oral immunization of animals with transgenic cherry tomatillo expressing HBsAg.
AIM: To investigate the expression of recombinant HBsAg (rHBsAg) in transgenic cherry tomatillo in order to explore the feasibility of producing HBV oral vaccine with cherry tomatillo by animal immune tests. METHODS: The recombinant plant expression vector containing HBsAg gene was constructed. Mediated with Agrobacterium tumefaciens, HBsAg gene was transferred into cotyledons of cherry tomatillo. Transformed cherry tomatillos were obtained through hygromycin delay-selection. Integrated DNA in transgenic cherry tomatillo was confirmed by hygromycin resistance selection, Gus detection, polymerase chain reaction (PCR) and dot blotting analysis. Antigenicity of rHBsAg was examined by ELISA and the immunogenicity of rHBsAg derived from transgenic cherry tomatillo tissues was confirmed by oral feed of transformed tissues to BALB/c mice primed with commercial HBV vaccines. Specific antibody titers in mice's serum were examined by ELISA every week. RESULTS: By far, 10 positive lines of transgenic cherry tomatillos containing HBsAg gene were obtained. Among different organs of the same transgenic cherry tomatillo, level of rHBsAg expressed in leaves was the highest with the yield up to 300ng/g fresh weight. And the rHBsAg expression level in fruits was about 10 ng/g fresh weight. In animal immune tests, oral delivery with transgenic tissues to mice primed with commercial vaccine instead of naive mice resulted in significant immune response. CONCLUSION: The result of this animal immune test indicated the rHBsAg derived from transgenic cherry tomatillo possessed normal immunogenicity. This work demonstrated the feasibility to generate oral immunogenic rHBsAg in transgenic cherry tomatillo, and would provide some experimental approach for the production of low-cost oral vaccines using transgenic cherry tomatillo in large scale.
Administration, Oral↗
Quantitative analysis of tumor-derived methylated p16INK4a sequences in plasma, serum, and blood cells of hepatocellular carcinoma patients.
PURPOSE AND EXPERIMENTAL DESIGN: Using real-time quantitative methylation-specific PCR (RTQ-MSP), we quantified methylated p16INK4a sequences and determined the fractional concentrations of circulating tumor DNA in plasma, serum, and peripheral blood cells collected preoperatively, intraoperatively, and postoperatively from 49 patients with hepatocellular carcinoma (HCC). RESULTS: RTQ-MSP was sufficiently sensitive to detect down to 10 genome-equivalents of methylated p16INK4a sequences. Quantitative MSP data were expressed in terms of the methylation index, which was the percentage of bisulfite converted unmethylated and methylated p16INK4a sequences that consisted of methylated p16INK4a sequences. Quantities of methylated p16INK4a sequences were detected in peripheral circulation of 80% (23 of 29) of HCC patients. No significant difference was seen in the detectability and concentrations of methylated p16INK4a sequences (range: 10-4046 genome-equivalents/ml) between preoperative plasma and serum samples from HCC patients. Preoperatively, the p16INK4a methylation indices ranged from 0.2 to 100% and from 0.012 to 0.075% in the patients' plasma and buffy coat samples, respectively. After surgical resection, the median p16INK4a methylation indices in plasma and buffy coat concordantly decreased 12- and 15-fold, respectively. These results demonstrated the clinical usefulness and effectiveness of peripheral blood RTQ-MSP for detecting and monitoring HCC after treatment. Furthermore, none of the intraoperative plasma samples and only two of the intraoperative buffy coat samples were p16INK4a methylation positive. CONCLUSIONS: Quantification of epigenetic changes in peripheral blood by RTQ-MSP is useful for the detection and monitoring of HCC.
Apoptosis↗
[Maturation regulation of dendritic cells pulsed with hepatocellular carcinoma cell soluble antigens].
OBJECTIVE: To research the maturation regulation of dendritic cells (DCs) pulsed with hepatocellular carcinoma (HCC) cell soluble antigens. METHODS: BCG HSP 70 was purified by SDS-PAGE electrophoresis and its biological activity was determined with ELISA. Phenotypes of DCs pulsed with antigens or with both antigens and BCG HSP 70 were analysed with flow cytometry. MTT assay was used to estimate the proliferation of self lymphocytes and the mixed lymphocyte reaction (MLR) of BCG HSP 70 primed DCs. RESULTS: The characteristics of DCs had changed after loaded with soluble antigens of HCC. There were about 10% DCs which had lost their specific markers. The expression levels of CD54, CD83, CD86 molecules and the stimulatory ability in allogeneic MLR decreased. However, after being activated by BCG HSP 70, the DCs pulsed with antigens could keep their special markers and the expression levels of CD54, CD83, CD86 molecules increased too. The stimulatory abilities in allogeneic MLR and proliferation of self lymphocytes also improved. CONCLUSION: This study shows that BCG HSP 70 can induce DCs pulsed with antigens maturation and improve their antigen-presenting ability, which may be a useful maturation inducer for dendritic cells.
Antigen Presentation↗
[Construction of shuttle expression vector and expression of thymosin alpha 1 in Synechococcus sp. PCC7942].
The shutter expression vector pPREUT was constructed from the plasmid pPRS-1 containing the endogenous small plasmid of Plectonema boryanum. The expression elements such as heat shock gene groESL promoter, foreign gene Ub-thymosin alpha 1, rbcS polyA terminator and Kanamycin resistance gene were all included. The shutter plasmid pPREUT was directly transferred into Synechococcus sp. PCC7942. The transformants were obtained through Kanamycin screening. Southern blotting analysis showed that the shutter plasmid have been transferred into Synechococcus sp. PCC7942. After induction by heat shock(42 degrees C) for 30 min, the foreign protein UB-T alpha 1 was expressed efficiently, which reached 7.5% of total amount protein.
Cyanobacteria↗
Atomic force microscopy of actin.
Atomic force microscope (AFM) is a powerful and novel tool to investigate the surface and submembranous structures of living cells under physiological conditions at high resolution, and has force measure at nano-Newton level. AFM can image and manipulate samples (single macromolecule, cells, and so on) at very high, sometimes atomic resolution by scanning a fine tip over the surface of interest and detecting physical interactions between the tip and sample. Actin is one of the most important proteins, involved extensively in various cell physiological activities and functions in eukaryotes. The present paper clarified the crucial significance and the enormous applicable prospects of AFM in modern biological sciences by reviewing the usage of AFM in actin study, which can be taken as a good example for expounding almost all functions that AFM owns, and can be utilized in life sciences exploration.
Actins↗
[Abnormality of p16 gene and its clinicopathological significance in non-small cell lung cancer].
OBJECTIVE: To investigate the ways of inactivation and expression of p16 gene mRNA and its protein as well, and their clinicopathological significance in non-small cell lung carcinomas (NSCLC). METHODS: Comparative- multiplex PCR, in situ hybridization, and immunohistochemistry were used to detect the promotor methylation status, mRNA, and protein expression in 64 cases of NSCLC, respectively. RESULTS: Promoter methylation of p16 gene was detected in 36 (56.3%) of 64 NSCLC cases. This positive result of methylation showed a negative correlation statistically with p16 protein expression by immunohistochemistry (P < 0.05). By in situ hybridization, 13 cases (20.3%) showed positive results for mRNA and all of these positive cases (13/13) had also a positive result by immunohistochemistry. Thirty-seven cases (57.8%) showed a negative immunohistochemical result. The metastatic rate of lymph nodes of those NSCLC patients with either promoter methylation or negative protein expression was higher (P = 0.038), and 3-year survival rate was lower statistically (P = 0.002). CONCLUSION: Dysfunction of p16 gene in NSCLC is mainly caused by promoter methylation, and patients with p16 gene dysfunction may have a poor prognosis.
Carcinoma, Non-Small-Cell Lung↗
[Analysis of 16 cases of uterine perforation during hysteroscopic electro-surgeries].
OBJECTIVE: To analyse the cause, diagnosis, treatment and preventive methods of uterine perforation resulting from hysteroscopic electro-surgeries. METHODS: Data of cases with uterine perforation were collected from 5 hospitals where overall 3,541 hysteroscopic electro-surgeries were done from May 1990 to July 2002. There were 1 468 transcervical resections of endometrium (TCRE), 797 cases of transcervical resection of myoma (TCRM), 783 cases of transcervical resection of endometrial polyp (TCRP), 189 cases of transcervical resection of uterine septa (TCRS), 112 cases of transcervical resection of uterine adhesion (TCRA) and 192 cases of transcervical removal of foreign body (TCRF). All operations were performed under B-ultrasonographic or laparoscopic monitoring. Cervical dilator stick was inserted into cervical canal or 200 micro g of misoprostol put in the posterior fornix the evening before operation. The procedures were done according to different indications and purposes. Cases of uterine perforation were divided into two groups: caused by approaching (entry-related) and by surgical instruments (technique-related). RESULTS: Totally sixteen cases (0.45%) of uterine perforation occurred. Seven cases occurred during cervical dilatation and 1 during hysteroscopy inserting lentry-related. Eight cases were technique-related caused by electrode. The incidences of uterine perforation of different operations were: TCRA 4.46% (5/112), TCRF 3.12% (6/192), TCRE 0.27% (4/1 468), TCRM 0.13% (1/797). TCRP and TCRS none. These 16 cases were all diagnosed during operations. 10 cases (62%) by B ultrasound and (or) laparoscopy, 6 cases (38%) by hysteroscopy and clinical features. 13 cases were complete uterine perforations, among them 2 were diagnosed by laparoscopic monitoring, 5 by B-ultrasonic monitoring, 4 by hysteroscopy and 2 by symptoms and B-ultrasound, 3 cases were incomplete uterine perforations in which 2 were diagnosed by laparoscopic monitoring and one by B-ultrasound monitoring. CONCLUSIONS: Half of uterine perforation cases were entry-related, so attention has to be paid to entry of Hegar or hysteroscopy (i.e., not dilate the cervix as possible and introduce the scope under direct vision). The other half were related to surgeons' experience and type of operation. TCRA and TCRF run more risks. B-ultrasound and (or) laparoscopy monitoring during hysteroscopic electro-surgery may help to prevent but not completely avoid uterine perforation.
Adult↗
Diagnosis, treatment and prognosis of thymoma: an analysis of 116 cases.
OBJECTIVE: To study the diagnosis and treatment of thymoma and to assess its prognostic factors. METHODS: The clinical data of 116 patients with thymoma were collected. A retrospective analysis was performed, by comparing the survival rate calculated by the Kaplan-Meier method with the rate of recurrence or metastasis. RESULTS: The standard posteroanterior and lateral chest radiographs were reliable means of detection of most thymomas. Myasthenia gravis was the most commonly paraneoplastic disease (25.0%, 29/116). The extensive radical resection was beneficial for reducing the rate of recurrence of stage I or stage II thymomas (chi(2) = 4.941, P = 0.0219). The survival time could be prolonged by postoperative radiotherapy and chemotherapy. There was a strong correlation between the clinical stage and the histological classification (according to MH classification), through which the invasive behavior of thymoma could be predicted (chi(2) = 19.76, P = 0.007, RR = 1.47). The 3- 5- and 10-year survival rates were 81.2%, 67.9%, and 40.5%, respectively. Statistical analysis showed a significant negative correlation between the stage and the survival rate (chi(2) = 29.73, P = 0.0000, RR = 0.15). CONCLUSION: The prognosis of thymoma depends mainly on the histological classification, clinical stage and multimodality treatment rather than on the paraneoplastic diseases.
Adolescent↗
[Study on clinical significances of the application of nasotracheal intubation guided with fiberbronchoscope to the postoperative patients with hypertensive intracerebral hemorrhage].
OBJECTIVE: To explore the clinical signification of the application of nasotracheal intubation guided with fiberbronchoscope to the postoperative patients with hypertensive intracerebral hemorrhage (HICH) who had respiration failure. METHODS: Ninety-four postoprative patients with HICH were divided into two groups, A group treated with nasotracheal intubation and B group treated with tracheal incision. The oxygen metabolism, pulmonary complication incidence of two groups were analyzed and compared. RESULTS: The basic oxygen metabolism and the need of mechanical ventilation in the cases with respiration failure were satisfying in two groups (both P>0.05). The mean reserved time of artificial airway in A group was(14.2+/-6.5)days, which in B group was(19.1+/-7.2)days (P<0.01). The occurent rates of combined infection in lung and double-infection were lower in A group than these in B group (56.3 percent vs. 91.3 percent, P<0.05; 10.4 percent vs. 39.1 percent, P<0.01; respectively), with their resident days shorter and prognosis better in A group than these in B group (P<0.05). CONCLUSION: The nasotracheal intubation guided with fiberbronchoscope has small trauma, lower incidence of complication, which may be considered as a better selection for the postoperative patients with HICH who had respiration failure.
Aged↗
[Analysis of factors related to mortality of critical patients with acute renal dysfunction].
OBJECTIVE: To investigate the high risk factors related to mortality rate of critical patients with acute renal dysfunction in intensive care unit (ICU). METHODS: The data of patients with acute renal dysfunction hospitalized in ICU of Peking union medical college hospital during August 1999 to May 2001 were retrospectively investigated and the high risk factors related to hospital mortality were analyzed. The statistical analysis methods were univariate and multivariate factors analysis. RESULTS: Among 1 218 critical patients hospitalized during August 1999 to May 2001, there were 149 patients with acute renal dysfunction. One hundred and thirty-five patients were investigated completely. Fifty-five (41 percent) died during the hospital stay. By univariate analysis, risk factors related to hospital mortality included acute physiology and chronic health evaluationII (APACHEII) score, the systemic inflammatory response syndrome, sepsis, severe sepsis, septic shock, chronic respiratory dysfunction, sepsis-related organ failure assessment (SOFA) score, acute respiratory dysfunction, coagulation dysfunction, central nerves system dysfunction, cardiovascular dysfunction, management of noradrenaline and continuous venovenous hemofiltration (CVVH). By multivariate analysis, only APACHEII score and management of noradrenaline and CVVH were determined as risk factors of hospital mortality independently. If the three factors were excluded, sepsis, central nerves system dysfunction, oliguria became the risk factors of mortality (all P<0.05). CONCLUSION: The prognosis of critical patients with the acute renal dysfunction correlate with degree of basic diseases. There include oliguresis, coagulation dysfunction, central nerves system dysfunction and sepsis. In spite of management of CVVH, acute renal dysfunction per se can affect the prognosis of the critical patients.
APACHE↗
[Degradation of fenpropathrin, phoxim and their mixture by soil microbes].
The degradation rates of fenpropathrin, phoxim and their mixture in un-sterilized soil were much quicker than those in sterilized soil, which indicated that soil microorganisms played a significant role in the degradation process in soil. The half-life (T0.5) in un-sterilized soil was 56.2 d for fenpropathrin, 57.8 d for mixed fenpropathrin, 48.2 d for phoxim, and 41.7 d for mixed phoxim. The corresponding half-life (T0.5) in sterilized soil was 135.1 d, 147.3 d, 123.6 d, and 126.2 d, respectively. There were no significant differences for degradation rates between single use and mixed use of fenpropathrin and phoxim.
Biodegradation, Environmental↗
[Multivariate analysis of recurrence in T1 bladder transitional cell carcinoma].
OBJECTIVE: To investigate the prognostic value of some clinicopathologic indexes and biologic tumor markers in predicting recurrence in T1 transitional cell carcinoma (TCC) of the bladder. METHODS: The expressions of p53, E-cadherin and VEGF of 75 patients with T1 primary bladder TCC were detected by streptabitin peroxidase (SP) immunohistochemical methods. The effects of clinicopathologic indexes and biologic tumor markers on recurrence were assessed by Kaplan-Meier and Cox proportional hazards model. RESULTS: The 1-, 3-, and 5-year recurrence- free survival rates were 68.0%, 45.3% and 20.9%. In Kaplan-Meier analysis, tumor mutifocality and the expression of p53, E-cadherin and VEGF were associated with recurrence. In multivariate analysis, the independent recurrence variables were tumor mutifocality, the expression of p53 and E-cadherin. CONCLUSION: Tumor mutifocality and the abnormal expression of p53 and E-cadherin are the variables that independently predict recurrence in T1 transitional cell carcinoma of the bladder.
Adult↗
[Impact of short-term neoadjuvant hormonal treatment on neuroendocrine differentiation in prostate carcinoma].
OBJECTIVE: To study the impact of short-term neoadjuvant hormonal treatment on neuroendocrine (NE) differentiation and the relation of NE differentiation and tumor regression. METHODS: The radical prostatectomy specimens and the biopsy specimens of the same 18 patients with prostate cancer were compared. The effect of hormonal treatment on NE-differentiation was evaluated by specific antibodies against chromogranin A (ChA) and serotonin (5-HT). RESULTS: The ChA-positive cell count was 3.2 x 10(-5)/microm(2) [(0-5.7) x 10(-5)/microm(2)] before hormonal treatment and 2.3 x 10(-5) microm(2)[(0-6.6) x 10(-5)/microm(2)] afterward (P > 0.05). For the proportion of NE-positive tumor, it was 7.0% (0%-14.9%) and 4.5% (0%-13.1%) (P > 0.05). No correlation existed between NE-differentiation and the neoadjuvant hormonal treatment. The NE cell density did not differ significantly between 12 non-/slightly regressive tumor foci and 6 highly regressive ones (P > 0.05). CONCLUSION: Short-term neoadjuvant hormonal therapy does not induce clonal propagation of NE cells. The degree of tumor regression following short-term neoadjuvant hormonal therapy is not correlated with the NE differentiation.
Aged↗
[Radiofrequency ablation therapy of hepatic neoplasms].
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Catheter Ablation↗