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Biomedical subjects

Zhi Chen

Publications and source records attributed to Zhi Chen.

At least 55 records · Page 3Linked to original sources

Association between TRAIL expression on peripheral blood lymphocytes and liver damage in chronic hepatitis B.

AIM: To explore a novel mechanism for tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), upregulation of CD4(+) and CD8(+) T lymphocytes participating in the patho-physiological process of chronic hepatitis B (CHB). METHODS: The levels of serum soluble TRAIL (sTRAIL), serum IFN-gamma and membrane-bound TRAIL expression on peripheral leucocytes from 58 CHB patients were examined by ELISA and flow cytometry respectively. The levels of TRAIL were compared with the baseline levels of 17 healthy controls, and correlation analysis was performed between ALT, TBIL, PT, morphological change in hepatic tissues, and serum IFN-gamma. RESULTS: The results showed that TRAIL levels on membranes of CD4(+), CD8(+) T cells in CHB patients were much higher than those in healthy controls (P<0.001), and were correlated with serum TBIL (r = 0.354, P = 0.008 for CD4(+) and r = 0.522, P = 0.000 for CD8(+), respectively), ALT (r = 0.393, P = 0.003 for CD8(+)), PT (r = 0.385, P = 0.004 for CD8(+)) and serum IFN-gamma level (r = 0.302, P = 0.011 for CD4(+) and r = 0.307, P = 0.009 for CD8(+)). On the contrary to membrane-bound TRAIL expression, serum level of sTRAIL was not correlated with that of TBIL and PT, though it was higher than that of the normal population and was positively correlated with serum HBeAg expression (r = 0.695, P = 0.001). CONCLUSION: The expression level of TRAIL on the membrane of lymphocytes was upregulated and associated with the liver injury in CHB patients. These findings suggest that upregulation of TRAIL expression may be induced by virus antigen and inflammatory cytokine IFN-gamma.

Adult↗

Chitosan-DNA microparticles as mucosal delivery system: synthesis, characterization and release in vitro.

BACKGROUND: Mucosal immunity is important to defense against dental caries. To enhance mucosal immunity, a DNA vaccine mucosal delivery system was prepared by encapsulating anticaries DNA vaccine (plasmid pGJA-P/VAX) in chitosan under optimal conditions and the characteristics of the microparticles was investigated. Furthermore, the release properties and protective action of microparticles for plasmid were studied in vitro. METHODS: Plasmid loaded chitosan microparticles were prepared by complex coacervation. Three factors, concentration of DNA, sodium sulfate, and the chitosan/DNA ratios in complexes [better expressed as N/P ratio: the number of poly nitrogen (N) per DNA phosphate (P)] influencing preparation were optimized by orthogonal test. The characteristics of microparticles were evaluated by scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM). DNA release rate of microparticles in similar gastro fluid (SGF) or similar intestinal fluid (SIF) at 37 degrees C was determined by ultraviolet spectrophotometry. RESULTS: High encapsulation efficiency (96.8%) was obtained with chitosan microparticles made under optimal conditions of 50 mmol/L Na2SO4, 200 microg/ml DNA and N/P ratio of 4. The size of particles was about 4 to 6 microm. The encapsulation process did not destroy the integrity of DNA. When incubated with SIL, after a release of about 10% in the first 60 minutes, no further DNA was released during the following 180 minutes. When incubated with SGL, the microparticles released a small burst (about 11%) in the first 60 minutes, and then slowly released at a constant, but different rate. CONCLUSIONS: These chitosan microparticles showed suitable characteristics in vitro for mucosal vaccination and are therefore a promising carrier system for DNA vaccine mucosal delivery.

Animals↗

DNA separation by EFFF in a microchannel.

This paper theoretically explores the application of electric field flow fractionation (EFFF) for the size-based separation of DNA strands in a microchannel. An axial electric field cannot separate DNA strands in solution because the electrical mobility of the strands is independent of the length. However, lateral electric fields coupled with an axial Poiseuille flow can separate the DNA strands of different sizes. By using regular perturbation analysis, we obtain the effective diffusivity and the mean velocity of the DNA molecules that are undergoing a pressure driven Poiseuille flow in a 2D channel in presence of a lateral electric field. The mean velocities and the dispersion coefficients are then utilized to determine the scaling for length of the channel and the time required for separation of DNA molecules in different parameter regimes. The results show that EFFF can separate DNA strands in the range of 10 kbp that differ in size by about 2.5 kbp in about half an hour in a 1 cm long channel. While DNA strands can be separated by EFFF, the performance of devices based on EFFF seems to be at best comparable to other techniques such as entropic trapping.

DNA↗

Quantifying signals with power-law correlations: a comparative study of detrended fluctuation analysis and detrended moving average techniques.

Detrended fluctuation analysis (DFA) and detrended moving average (DMA) are two scaling analysis methods designed to quantify correlations in noisy nonstationary signals. We systematically study the performance of different variants of the DMA method when applied to artificially generated long-range power-law correlated signals with an a priori known scaling exponent alpha(0) and compare them with the DFA method. We find that the scaling results obtained from different variants of the DMA method strongly depend on the type of the moving average filter. Further, we investigate the optimal scaling regime where the DFA and DMA methods accurately quantify the scaling exponent alpha(0) , and how this regime depends on the correlations in the signal. Finally, we develop a three-dimensional representation to determine how the stability of the scaling curves obtained from the DFA and DMA methods depends on the scale of analysis, the order of detrending, and the order of the moving average we use, as well as on the type of correlations in the signal.

Journal Article↗

Risk of childhood germ cell tumors in association with parental smoking and drinking.

BACKGROUND: The etiology of childhood germ cell tumors (GCT) is not well understood. The Children's Oncology Group conducted the largest case-control study of childhood GCT to investigate whether parental exposures to smoking and alcohol contributed to the disease. METHODS: Cases included 274 children with GCT diagnosed between January 1, 1993 and December 31, 2001 who were age <15 years. Controls (n=421) were selected by random digit dialing and were frequency matched based on gender, age (+/-1 year), and geographic area. Exposure information was collected from subjects' parents using independent telephone interviews and self-administrated questionnaires. RESULTS: No association was found between parental smoking or drinking alcohol and risk of childhood GCT (for smoking: odds ratio [OR]=1.0, 95% confidence interval [95% CI], 0.8-1.3 and OR = 1.2, 95% CI, 0.9-1.5, for mothers and fathers, respectively; for drinking: OR=0.9, 95% CI, 0.7-1.2 and OR=1.0, 95% CI, 0.8-1.3, for mothers and fathers, respectively). No significant trend was observed for length of maternal exposure to passive smoking during the index pregnancy and GCT risk (for total subject: P=0.77; boys: P=0.52; girls: P=0.93). CONCLUSIONS: The authors found no evidence that childhood GCT was related to prenatal exposure to parental cigarette smoking, alcohol drinking, and maternal passive smoking.

Adolescent↗

Separation of charged colloids by a combination of pulsating lateral electric fields and poiseuille flow in a 2D channel.

Separation of colloidal particles of different sizes is becoming increasingly important due to rapid developments in the area of proteomics, genetic engineering, drug discovery, etc. In particular, there is a need to accomplish these separations on a microscale in 'lab-on-a-chip' devices. In this paper, we propose a new method for accomplishing separation of charged colloids of different sizes in a microchannel. This method involves a combination of pulses of lateral electric fields and Poiseuille flow in the axial direction. We develop a model for this separation technique and obtain closed form solutions for the mean velocity and the dispersion coefficient for a pulse of molecules introduced into the channel. These expressions are then utilized to determine the channel length and the separation time. For reasonable value of design constants, the proposed technique can separate molecules of different sizes that have diffusivities of 10(-10) and 0.5 x 10(-10) m2/s in 15.7 s in a 3.7 mm long channel. The length and the time increase to 5.45 cm and 231 s if the ratio of the diffusivities is reduced from 2 to 1.2, i.e., the latter diffusivity is increased to 0.835 x 10(-10) m2/s, while keeping all the other parameters the same. If the diffusivities are about 10(-9) m(2)/s, the length and the time for separation are 1 cm and 17.5 s for D1/D2=2, and 16 cm and 269 s for D1/D2=1.2.

Journal Article↗

Inhibition of hepatitis B virus surface antigen expression by small hairpin RNA in vitro.

AIM: To explore the anti-hepatitis B virus effect of RNA interference (RNAi) using small hairpin RNA (shRNA) expression vector. METHODS: Hepatitis B virus surface antigen green fluorescent protein (HBs-GFP) fusion vector and shRNA expression vectors were constructed and cotransfected transiently into HepG2 cells. mRNAs extracted from HepG2 cells were detected by real-time PCR. Fluorescence of HBs-GFP protein was detected by fluorescence-activated cell sorting (FACS). The effective shRNA expression vector was transfected into HepG2.2.15 cells. HBsAg and HBeAg in HepG2.2.15 cells were analyzed by radioimmunoassay (RIA) method. RESULTS: FACS revealed that shRNA targeting at HBsAg reduced the GFP signal by 56% compared to the control. Real-time PCR showed that HBs-GFP mRNA extracted from HepG2 cells cotransfected with pAVU6+27 and HBs-GFP expression plasmids decreased by 90% compared to the empty vector control. The expressions of HBsAg and HBeAg were also inhibited by 43% and 64%, respectively. CONCLUSION: RNAi using shRNA expression vector can inhibit the expression of HBsAg, providing a fresh approach to screening the efficient small interfering RNAs (siRNAs).

Carcinoma, Hepatocellular↗

Effect of nonlinear filters on detrended fluctuation analysis.

When investigating the dynamical properties of complex multiple-component physical and physiological systems, it is often the case that the measurable system's output does not directly represent the quantity we want to probe in order to understand the underlying mechanisms. Instead, the output signal is often a linear or nonlinear function of the quantity of interest. Here, we investigate how various linear and nonlinear transformations affect the correlation and scaling properties of a signal, using the detrended fluctuation analysis (DFA) which has been shown to accurately quantify power-law correlations in nonstationary signals. Specifically, we study the effect of three types of transforms: (i) linear ( y(i) =a x(i) +b) , (ii) nonlinear polynomial ( y(i) =a x(k)(i) ) , and (iii) nonlinear logarithmic [ y(i) =log ( x(i) +Delta) ] filters. We compare the correlation and scaling properties of signals before and after the transform. We find that linear filters do not change the correlation properties, while the effect of nonlinear polynomial and logarithmic filters strongly depends on (a) the strength of correlations in the original signal, (b) the power k of the polynomial filter, and (c) the offset Delta in the logarithmic filter. We further apply the DFA method to investigate the "apparent" scaling of three analytic functions: (i) exponential [exp (+/-x+a) ] , (ii) logarithmic [log (x+a) ] , and (iii) power law [ (x+a)(lambda) ] , which are often encountered as trends in physical and biological processes. While these three functions have different characteristics, we find that there is a broad range of values for parameter a common for all three functions, where the slope of the DFA curves is identical. We further note that the DFA results obtained for a class of other analytic functions can be reduced to these three typical cases. We systematically test the performance of the DFA method when estimating long-range power-law correlations in the output signals for different parameter values in the three types of filters and the three analytic functions we consider.

Algorithms↗

Making a tooth: growth factors, transcription factors, and stem cells.

Mammalian tooth development is largely dependent on sequential and reciprocal epithelial-mesenchymal interactions. These processes involve a series of inductive and permissive interactions that result in the determination, differentiation, and organization of odontogenic tissues. Multiple signaling molecules, including BMPs, FGFs, Shh, and Wnt proteins, have been implicated in mediating these tissue interactions. Transcription factors participate in epithelial-mesenchymal interactions via linking the signaling loops between tissue layers by responding to inductive signals and regulating the expression of other signaling molecules. Adult stem cells are highly plastic and multipotent. These cells including dental pulp stem cells and bone marrow stromal cells could be reprogrammed into odontogenic fate and participated in tooth formation. Recent progress in the studies of molecular basis of tooth development, adult stem cell biology, and regeneration will provide fundamental knowledge for the realization of human tooth regeneration in the near future.

Animals↗

Evaluation of antiviral activity of compounds isolated from Ranunculus sieboldii and Ranunculus sceleratus.

Nineteen compounds isolated from Ranunculus sieboldii and Ranunculus sceleratus were tested for inhibitory effects on hepatitis B virus (HBV) and Herpes simplex virus type-1 (HSV-1). The results showed that apigenin 4'- O- alpha-rhamnopyranoside, apigenin 7- O- beta-glucopyranosyl-4'- O- alpha-rhamnopyranoside, tricin 7- O- beta-glucopyranoside, tricin, and isoscopoletin possessed inhibitory activity against HBV replication. Protocatechuyl aldehyde exhibited an inhibiting activity on HSV-1 replication. It is therefore suggested that further investigations on these bioactive compounds might be needed to discover and develop new antiviral agents.

Antiviral Agents↗

periostin null mice exhibit dwarfism, incisor enamel defects, and an early-onset periodontal disease-like phenotype.

Periostin was originally identified as an osteoblast-specific factor and is highly expressed in the embryonic periosteum, cardiac valves, placenta, and periodontal ligament as well as in many adult cancerous tissues. To investigate its role during development, we generated mice that lack the periostin gene and replaced the translation start site and first exon with a lacZ reporter gene. Surprisingly, although periostin is widely expressed in many developing organs, periostin-deficient (peri(lacZ)) embryos are grossly normal. Postnatally, however, approximately 14% of the nulls die before weaning and all of the remaining peri(lacZ) nulls are severely growth retarded. Skeletal analysis revealed that trabecular bone in adult homozygous skeletons was sparse, but overall bone growth was unaffected. Furthermore, by 3 months, the nulls develop an early-onset periodontal disease-like phenotype. Unexpectedly, these mice also show a severe incisor enamel defect, although there is no apparent change in ameloblast differentiation. Significantly, placing the peri(lacZ) nulls on a soft diet that alleviated mechanical strain on the periodontal ligament resulted in a partial rescue of both the enamel and periodontal disease-like phenotypes. Combined, these data suggest that a healthy periodontal ligament is required for normal amelogenesis and that periostin is critically required for maintenance of the integrity of the periodontal ligament in response to mechanical stresses.

Animals↗

Fabrication of a one-dimensional array of nanopores horizontally aligned on a Si substrate.

A one-dimensional array of nanopores horizontally aligned on a silicon substrate was successfully fabricated by anodic aluminum oxidation (AAO) using a modified two-step procedure. SEM pictures show clear nanostructures of well-aligned one-dimensional nanopore arrays without cracks at the interfaces of the sandwiched structures. The processes are compatible with the planar silicon integrated circuit processing technology, promising for applications in nanoelectronics. The formation mechanism of a single nanopore array on Si substrates was also discussed.

Aluminum Oxide↗

In vitro antiviral activity of three enantiomeric sesquiterpene lactones from Senecio species against hepatitis B virus.

Three enantiomeric sesquiterpene lactones were isolated under the bioguidance of the suppression of hepatitis B virus (HBV) surface antigen (HBsAg) from Senecio species, a widely distributed Chinese medicinal herb traditionally used for the treatment of hepatitis B, dermatosis and inflammation. The anti-HBV activity of the purified compounds was measured; all of them showed suppressive activity on the expression of HBsAg and HBV e antigen (HBeAg) in the HepG2.2.15 cell line. Real-time quantitative PCR analysis showed that the studied compounds decreased the number of infectious virions released, but did not inhibit the intracellular HBV DNA. The results suggest that enantiomeric sesquiterpene lactones may possess the potential to work synergistically with other antiviral compounds for the treatment of HBV infection.

Antiviral Agents↗

Construction of a eukaryotic expression plasmid of Humanin.

OBJECTIVE: To construct a eukaryotic expression plasmid pcDNA3.1(-)-Humanin. METHODS: The recombinant plasmid pGEMEX-1-Humanin was digested with restriction endonucleases BamH I and Hind III and the Humanin gene fragments, about 100 bp length, were obtained. Then the Humanin gene fragments were inserted into eukaryotic expression vector pcDNA3.1(-) and the recombinant plasmids pcDNA3.1(-)-Humanin were identified by sequencing. RESULTS: Recombinant plasmid DNA successfully produced a band which had the same size as that of the Humanin positive control. The sequence of recombinant plasmids accorded with the Humnain gene sequence. CONCLUSIONS: A eukaryotic expression plasmid of Humanin was successfully constructed.

Base Sequence↗

Construction and characterization of a cDNA library from human liver tissue with chronic hepatitis B.

OBJECTIVE: To construct a cDNA library from human liver tissue with chronic hepatitis B and check its quality for investigating the expression level of liver tissue infected by hepatitis B virus. This will then be used to find the relevant genes and interesting proteins associated with the development of hepatitis B. METHODS: The total RNA from liver tissue with chronic hepatitis B was extracted and the mRNA was purified using TRIZOL method. Switching mechanism at 5' end of the RNA transcript (SMART) technique and CDS III/3' primer were used for first-strand cDNA synthesis. Long distance polymerase chain reaction (LD PCR) was then used to synthesize the double-strand cDNA that was then digested by Sfi I and fractionated by CHROMA SPIN-400 column. The longer than 0.4 kb cDNAs were collected and ligated to lambdaTriplEx2 vector. Then lambda phage packaging reaction and library amplification were performed. The qualities of both unamplified and amplified cDNA libraries were strictly checked by conventional titer determination. Fourteen plaques were randomly picked and tested using PCR with universal primers derived from the sequence flanking the vector. RESULTS: The titers of unamplifed and amplified libraries were 1.94 x 10(6) pfu/ml and 1.49 x 10(9) pfu/ml respectively. The percentages of recombinants from both libraries were 98.15% in unamplified library and 98.76% in amplified library. The lengths of the inserts were 1.23 kb in average, 1-2 kb in 64.29%, and 0.5-1.0 kb in 35.71%. CONCLUSION: A high quality cDNA library from human liver tissue with chronic hepatitis B was successfully constructed.

DNA, Complementary↗

Proapoptotic and pronecrosis effect of different truncated hepatitis C virus core proteins.

OBJECTIVE: To study the roles of different truncated hepatitis C virus (HCV) core proteins (CORE) in the pathogenesis of HCV persistent infection and hepatocellular carcinoma (HCC) and to assess intracellular localization in transiently transfected cells. METHODS: Seven truncated GFP (green fluorescent protein)-CORE fusion protein expression plasmids were constructed, which contained HCV CORE sequences derived from tumor tissues (BT) and non-tumor tissues (BNT) from one patient infected with HCV. Amino acid (aa) lengths were BT: 1-172 aa, 1-126 aa, 1-58 aa, 59-126 aa, 127-172 aa; BNT: 1-172 aa and C191: 1-172 aa respectively. Subcellular localization of CORE-GFP was analyzed by con-focal laser scanning microscope. Apoptosis and necrosis were quantified by flow cytometry. RESULTS: Different truncated CORE-GFP localized mainly in the cytoplasm, but nuclear staining was also observed. HCV CORE could induce apoptosis and necrosis, and different truncated COREs could induce cell apoptosis and necrosis at different levels. Among the same length 1-172 aa of BT, BNT and C191, the cell apoptosis and necrosis percentage of BT is highest, and C191 is the lowest (BT>BNT>C191). To the different fragment COREs of BT, N-terminal of CORE induced apoptosis and necrosis higher, compared with that of C-terminal (1-172 aa>1-126 aa>1-58 aa>127-172 aa>59-126 aa). CONCLUSION: These results suggest HCV CORE could induce apoptosis and necrosis of cells, which might play an important role in the pathogenesis of HCV persistent infection and HCC and the different CORE domains of different HCV quasi-species might have some difference in their pathogenesis.

Apoptosis↗

Immunomodulatory function of orally administered thymosin alpha1.

OBJECTIVE: To investigate the immunological function of a yeast expression system for thymosin alpha1 (Talpha1). METHODS: A constructed Talpha1 yeast expression system was used to investigate the immunological function of orally administered Talpha1. Dried yeast containing three different concentration of Talpha1 was fed to normal Balb/c mice and other Balb/c mice whose immunities were inhibited in advance by cyclophosphamide. Synthesized Talpha1 peptide was used as positive control and dried yeast with empty plasmid was used as negative control. CD4(+) and CD8(+) levels were detected by flow cytometry assay. TNF-alpha, IFN-gamma, IL-2, IL-6 and IL-10 levels were detected by liquid chip. RESULTS: In normal Balb/c mice or immune inhibition Balb/c mice, CD8(+) levels were significantly increased. Especially in immune inhibition Balb/c mice, CD8(+) levels in synthesized Talpha1 group (18.77%+/-4.72%), small dose group (13.48%+/-6.17%) and large dose group (22.74%+/-1.09%) were significantly higher than that in empty yeast control group (7.49%+/-2.14%). CONCLUSION: Orally administered Talpha1 has its certain immunomodulatory function.

Administration, Oral↗