[Diversity of clinical manifestation of chronic hepatitis C].
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Cryoglobulinemia↗
Biomedical subjects
Jun Cheng
Publications and source records attributed to Jun Cheng.
[Relations of methionine synthase gene variation with congenital heart disease].
OBJECTIVE: Methionine synthase (MS) is the key enzyme in the homocysteine metabolism. To investigate the relations of MS gene variation with occurrence of congenital heart disease (CHD). METHODS: 186 CHD patients (0-31 years old) were selected as case group and 103 normal population as control. For all subjects the gene polymorphism at MS A2756G locus was analysed by PCR-RFLP method, and the serum folic acid/vitamin B12 levels were detected by radio-immunity assay. RESULTS: The heterozygotes (+/-) were detected in the subjects but without homozygotes (+/+). In control group the frequencies of (+/-) genotype and (+) allele were 10.7% and 5.3%, lower than Caucasian and Japanese population. In case group the frequencies of (+/-) genotype and (+) allele were 9.1% and 4.6%, without significantly different from control. The odds ratio of (+/-) genotype was 0.84 (0.35, 2.01). The genotype distributions in different types of CHD were also not apparently different with control. The serum vitamin B12 level was decreased in case group compared with control (336.66 pmol/L vs 465.72 pmol/L, P > 0.05), but the serum folic acid level no different. Also there were not significant difference for folic acid/vitamin B12 levels between different genotypes in case group. CONCLUSION: The results indicated that there was not apparent association between MS gene A2756G locus variation with CHD and serum folic acid/vitamin B12 levels. It need further investigations.
5-Methyltetrahydrofolate-Homocysteine S-Methyltran↗
Polymorphism of methionine synthase gene in nuclear families of congenital heart disease.
OBJECTIVE: To investigate the relation of methionine synthase (MS) gene variation with congenital heart disease (CHD) phenotype. METHODS: One hundred and ninety three CHD patients (94 males and 99 females) and their biological parents (nuclear families) in Liaoning Province were selected as the case group, and another 104 normal persons (60 males and 44 females) and their parents without family history of birth defects as the control group. For all subjects the polymorphism of MS gene A2756G locus was examined by PCR-RFLP method. RESULTS: In offspring of the control group the frequencies of MS genotype (+/-) and allele (+) were 10.7% and 5.3%, without existence of homozygote. The MS genotype distribution and allele frequencies of CHD patients and their mothers were not significantly different from the control (P > 0.05). The frequency of allele (+) in case fathers (5.0%) was apparently lower than that in the control (9.1%, P = 0.060), and the odds ratio (OR) was 0.53 (95% CI: 0.25-1.09). There was no difference in parents' genotype combination between the two groups, and in genotype distribution among different types of CHD. Analysis of genetic transmission indicated that mutation allele (+) existed transmission disequilibrium in CHD nuclear families. The percentage of allele (+) transmitted from parents was lower than that allele (-) with OR 0.26 (95% CI: 0.11-0.60). CONCLUSION: MS gene variation in parents is associated with occurrence of CHD in offspring, and mutation allele (+) in parents may be related with the decrease of CHD risk in offspring.
5-Methyltetrahydrofolate-Homocysteine S-Methyltran↗
[Epstein-Barr virus infection in lymphoepithelial carcinoma of salivary glands in Sichuan Chinese].
OBJECTIVE: To determine the prevalence of Epstein-Barr virus (EBV) infection in lymphoepithelial carcinoma (LEC) of salivary glands in Sichuan Chinese and discuss the role of EBV infection in pathogenesis of LEC. METHODS: Paraffin sections of 16 cases of LEC and 4 cases of benign lymphoepithelial lesion (BLEL) of salivary glands, 4 cases of non-specific chronic sialadenitis were examined by in situ hybridization for EBV encoded small RNA-1 (EBER-1) using 30-base synthesized oligonucleotide probe. RESULTS: Strong signals for EBER-1 were obtained in most of tumor cell nuclei in all the specimens of LEC (16/16); in contrast, there were no significant signals observed in the examination of 4 cases of BLEL and 4 cases of non-specific chronic sialadenitis. CONCLUSION: The results indicate that EBV plays an important role in the pathogenesis of LEC of salivary glands in Sichuan Chinese.
Adult↗
[Mutual interaction between hepatitis B virus antigen and metallothionein].
OBJECTIVE: To screen and identify the protein interacting with HBV antigen in hepatocytes. Then investigate the biological functions of hepatitis B virus antigen in the pathogenesis of hepatitis B and seek effective methods to prevent and treat it. METHODS: The yeast two-hybrid system-3 technique was used to construct HBV PreS2, HBeAg, HBcAg, HBxAg bait plasmids. The bait plasmids transformed the yeast AH109 and expressed themselves in it. After being identified by SDS-PAGE and Western blot, the AH109 yeast was mated with yeast Y187 containing liver cDNA library plasmid in 2 x YPDA medium to form diploid yeast and was then plated on synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) and synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) containing x-alpha-gal for screening. Plasmids of blue colonies were extracted and transformed into Escherichia coli, then analyzed by DNA sequencing and bioinformatics. To further prove the interaction between HBV antigen and metallothionein, translation was performed by using reticulocyte lysate and coimmunoprecipitation was displayed in vitro. RESULTS: Genes coding for HBV antigen binding protein were successfully cloned and metallothionein was found in that protein. The interaction between HBeAg, HBcAg and HBxAg and metallothionein were further proved by coimmunoprecipitation in vitro. CONCLUSION: The interaction between HBV antigen and metallothionein indicates that metallothionein may participate in the pathogenesis of hepatitis B
Hepatitis B↗
[RNA interference technique in the study of gene expression regulation].
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Gene Expression Regulation↗
[Epstein-Barr virus infection in benign lymphoepithelial lesions with malignant transformation of salivary glands].
OBJECTIVE: To examine the role of Epstein-Barr virus (EBV) infection in benign lymphoepithelial lesions with malignant transformation of salivary glands. METHODS: 2 cases benign lymphoepithelial lesions with focal malignant transformation (BLEL-FMT), 14 cases of malignant lymphoepithelial lesion (MLEL) and 4 cases of benign lymphoepithelial lesion (BLEL) of salivary glands, 4 cases of nonspecific chronic sialadenitis were examined. Sections were cut from the paraffin blocks and performed in situ hybridization (ISH) for EBV encoded small RNA-1 (EBER-1) using 30-base synthesized oligonucleotide probe and polymerase chain reaction (PCR) for BamHI-W regein of EBV DNA (123 bp). RESULTS: In ISH, strong signals for EBER-1 were obtained in most of epithelial cell nuclei in examined specimens of BLEL-FMT, no matter in benign or malignant areas and in all specimens of MLEL (14/14), PCR demonstrated a DNA fragment of 123 kbp in 2 cases of BLEL-FMT and 12 cases of MLEL (12/14). In contrast, there were no significant ISH/PCR signals in examined BLEL and nonspecific chronic sialadenitis. CONCLUSIONS: A few cases of MLEL may arise from BLEL and EBV may plays an important role in the pathogenesis of BLEL-FMT of salivary glands.
Epstein-Barr Virus Infections↗
[Effect of clozapine and risperidone on serum cytokine levels in patients with first-episode paranoid schizophrenia].
OBJECTIVE: To study the effects of clozapine and risperidone on serum cytokine levels in patients with first-episode paranoid schizophrenia, and explore the role of the cytokines in the psychopathological basis of the illness. METHOD: Fifty-eight patients with first-episode paranoid schizophrenia were treated with either clozapine or risperidone, and before and at the end of the 4th, 8th weeks and 6th months after the medication respectively, the serum levels of interleukin (IL)-6, 2, 18,and tumor necrosis factor (TNF)alpha were measured with enzyme-linked immunosorbent assay (ELISA). The Positive and Negative Syndrome Scale (PANSS) was used to evaluate the psychotic symptoms. RESULT: In patients treated with risperidone, the levels of serum IL-6 and IL-2 after 4 weeks, TNFalpha after 8 weeks, and IL-18 after 6 months were all significantly lowered in comparison with the pretreatment levels (P<0.01 or 0.05). In clozapine group, the levels of IL-2 after 4 weeks and IL-6 and IL-18 after 6 months were lowered significantly (P<0.01 or 0.05). Before the medication, serum IL-6 level was positively correlated with Positive Syndrome scores (r=0.386, P<0.01), IL-2 with the total score and Positive Syndrome scores (r=0.338, 0.305; P<0.01, 0.05), and TNFalpha with the total score (r=0.283, P<0.05). The changes of IL-2 and IL-6 after 8 weeks was positively correlated with the change of Positive Syndrome scores (r=0.268, 0.375; P<0.05, 0.01). Six months after the medication, the change in IL-6 and TNFalpha levels was positively correlated with the change of total score (r=0.365, 0.362; P<0.05). Before treatment, IL-6 was positively correlated with IL-2 levels (r=0.356, P<0.01), and TNFalpha with IL-18 levels (r=0.291, P<0.05). TNFalpha was positively correlated with IL-6 levels (r=0.332, P<0.01) 8 weeks after the medication. The changes in IL-6 was positively correlated with the those in IL-2 levels 6 months after the medication (r=0.391, P< 0.05). CONCLUSION: Clozapine and risperidone have similar immunosuppression actions and may affect serum IL-6 levels in patients with paranoid schizophrenia, in the psychopathology of which the cytokines play their roles of various importance.
Adolescent↗
Mutational events in LMP1 gene of Epstein-Barr virus in salivary gland lymphoepithelial carcinomas.
It is still unknown what kinds of roles Epstein-Barr virus (EBV) infection that are highly specific to salivary gland lymphoepithelial carcinomas (LECs) play in their tumorigenesis. To clarify the significance of EBV in LECs, we paid particular attention to the LMP1 gene, which is responsible for triggering several pathways for activating transcription factors. Sixty-one cases of EBV positive LECs confirmed by PCR and in-situ hybridization were collected from various areas of the world and studied immunohistochemically for latent membrane protein-1. Furthermore, PCR for the LMP1 carboxyl (C)-terminus region was performed, and the PCR products were sequenced for detection of other mutational events. LMP1 gene products were immunohistochemically demonstrated in 51% of the cases, while PCR amplification of the LMP1 gene was successful in 41 cases (67%). Among them, a 30 bp deletion in the C-terminus of the LMP1 gene, which had been shown to be characteristic to EBV in Chinese nasopharyngeal carcinomas, was found in 20 cases (32%). Most of them were from Guangzhou, Chengdu and Taiwan, while most of the cases from Shanghai and other areas exhibited no 30 bp deletion. In addition, several point mutations including codon 338 of LMP1 were commonly shared by the cases with or without the 30 bp deletion. These results indicate that there are 2 major genomic variations of EBV infecting salivary gland LECs. The frequent mutational events in the C-terminus in addition to the 30 bp deletion also seem to be critical for the pathogenesis because such mutational events may possibly promote cellular proliferation.
Carcinoma, Squamous Cell↗
Vascular endothelial cell participation in formation of lymphoepithelial lesions (epi-myoepithelial islands) in lymphoepithelial sialadenitis (benign lymphoepithelial lesion).
Lymphoepithelial lesions (LELs, or epi-myoepithelial islands) in lymphoepithelial sialadenitis (LESA, or benign lymphoepithelial lesion) of the salivary gland are known to be mainly composed of duct epithelial cells. However, other constituent cells are poorly characterized. Formalin-fixed paraffin sections obtained from six surgical specimens of LESA were examined using immunohistochemistry for cytoskeletal proteins, inflammatory cells, vascular endothelial cells, and extracellular matrix (ECM) molecules as well as by in situ hybridization for ECM molecules. In addition to keratin-immunopositive (+) duct-like epithelial cells, there were CD31/CD34+ vascular endothelial cells-which were either scattered in a singular fashion, in formed sheets, or in tubular structures-, CD20+ B lymphocytes, CD45RO+ T lymphocytes, and CD68 macrophages in the LELs. ECM molecules, such as heparan sulfate proteoglycan and tenascin, were immunolocalized in hyaline materials in the LELs. Their mRNAs were demonstrated mainly in endothelial cells and, to a lesser extent, in lympho-monocytic cells around hyaline materials, but were not as evident in epithelial constituent cells of LELs. The results indicate that endothelial cells as well as inflammatory cells are important constituents of the LELs, and the hyaline ECM cores mainly result from the intra-LEL angiogenesis by endothelial cells with the assistance of inflammatory cells. This intra-LEL vasculature seems to support regeneration and proliferation of salivary epithelial remnant cells.
Antigens, CD↗
Use of disposable gold working electrodes for cation chromatography-integrated pulsed amperometric detection of sulfur-containing amino acids.
We have prepared disposable thin-film gold working electrodes on polymeric substrates. Our microfabrication process allows for inexpensive and reproducible mass production of such electrodes. We utilize this new type of electrode in flow-through electrochemical cells to replace the conventional non-disposable gold working electrodes for integrated pulsed amperometric detection (IPAD) of compounds separated by high-performance cation-exchange chromatography. Using two S-containing amino acids (homocysteine and cysteine) as test compounds, we have modified a previously reported waveform for optimum performance with disposable gold electrodes. With the help of the same two test substances we have characterized the analytical performance of disposable gold electrodes under the new conditions. Compared to non-disposable working electrodes, the disposable working electrodes generated equal or better results in the limit of detection, linearity of calibration and reproducibility. When used with a new IPAD waveform, the disposable electrodes functioned reproducibly for 3 days. At the end of the specified usage period of 3 days, the disposable electrodes are simply replaced. Reconditioning by polishing is thus no longer required.
Calibration↗
Inhibition of fast sodium current in rabbit ventricular myocytes by protein tyrosine kinase inhibitors.
The present study investigated the effects of protein tyrosine kinase inhibitors on the fast sodium current ( I(Na)) in rabbit ventricular myocytes. Single rabbit ventricular myocytes were isolated enzymatically using Langendorff perfusion. I(Na) was recorded using the whole-cell patch-clamp technique at room temperature. The protein tyrosine kinase inhibitors genistein, AG957, ST638, and PP2 reversibly inhibited I(Na) in a concentration-dependent manner. At a test pulse potential of -30 mV, genistein (n=7) inhibited I(Na) by 37.7+/-3.2%, 53.4+/-2.5%, and 71.8+/-2.7% at concentrations of 15, 50, and 100 microM, respectively, without changing the voltage dependence of activation, while 100 microM AG957, 100 microM ST638, and 30 microM PP2 inhibited I(Na) by 38.7+/-2.4, 35.8+/-3.4, and 21.1+/-3.9%, respectively. Genistein (100 microM) and AG957 (100 microM) shifted the voltage for half-maximal inactivation of I(Na) from -76.7+/-2.0 mV (n=10) in control to -88.37+/-2.6 mV (n=6, P<0.05), and -82.9+/-1.7 (n=4, P<0.05), respectively, without changing the slope factor. Genistein and AG957 also significantly prolonged the time course of I(Na) recovery from inactivation. Daidzein and PP3, inactive analogs of genistein and PP2, respectively, did not inhibit I(Na) significantly. We conclude that protein tyrosine kinase signaling pathways may play an important role in regulation of I(Na) in cardiac myocytes.
Animals↗
Screening and characterization of human single-chain Fv antibody against beta-amyloid peptide 40.
Expression of single-chain variable fragment antibody on the surface of filamentous bacteriophage is widely used to make antibodies with pre-defined specificities. Using direct selection on solid phase-bound amyloid-beta 40 peptide, which is a main pathogenic feature of Alzheimer's disease, we obtained single-chain Fv antibody with specific binding activity. The binding epitope of the antibody is located between amino acids 1 and 16 of the peptide antigen. DNA sequencing showed that the gene coding for the single chain Fv antibody consists of 768 bp, and the complementarity-determining regions were deduced.
Amino Acid Sequence↗
Development and characterization of microfabricated disposable gold working electrodes for high-performance ion chromatography and integrated pulsed amperometric detection.
We have developed a new type of microfabricated thin-film electrode on polymeric substrates. The microfabrication process allows for inexpensive and reproducible mass production of disposable working electrodes for high-performance ion chromatography and integrated pulsed amperometric detection (IPAD). These microfabricated electrodes are disposable and have been optimized for use in flow-through low-dead-volume electrochemical cells. The analytical performance of microfabricated gold electrodes was characterized with the help of the IPAD method for amino acid detection under alkaline conditions required for anion-exchange separations. When used with a new optimized six-potential IPAD waveform, the electrodes functioned properly for weeks. Compared to nondisposable working electrodes, the disposable working electrodes generated equal or better results in the limit of detection, linearity of calibration, and reproducibility. Disposable electrodes make it possible to avoid polishing and reconditioning, which are required with nondisposable electrodes.
Journal Article↗
Targeted disruption of pyrin, the FMF protein, causes heightened sensitivity to endotoxin and a defect in macrophage apoptosis.
Familial Mediterranean fever (FMF) is an inherited disorder characterized by recurrent episodes of fever and inflammation. Most patients with FMF carry missense mutations in the C-terminal half of the pyrin protein. To study the physiologic role of pyrin, we generated mice expressing a truncated pyrin molecule that, similar to FMF patients, retains the full PYRIN domain. Bacterial lipopolysaccharide (LPS) induces accentuated body temperatures and increased lethality in homozygous mutant mice. When stimulated, macrophages from these mice produce increased amounts of activated caspase-1 and, consequently, elevated levels of mature IL-1beta. Full-length pyrin competes in vitro with caspase-1 for binding to ASC, a known caspase-1 activator. Apoptosis is impaired in macrophages from pyrin-truncation mice through an IL-1-independent pathway. These data support a critical role for pyrin in the innate immune response, possibly by acting on ASC, and suggest a biologic basis for the selection of hypomorphic pyrin variants in man.
Alternative Splicing↗
Jaw bone remodeling at the invasion front of gingival squamous cell carcinomas.
BACKGROUND: It is still unknown how jaw bone remodeling occurs at actual invasion sites of oral squamous cell carcinomas. Since there is no other human carcinomas which make a direct invasion of the bone, gingival carcinomas are valuable examples. METHODS: Twelve surgical specimens of gingival squamous cell carcinoma were examined histopathologically and immunohistochemically for remodeling of bone and its surrounding tissue. RESULTS: Three types of bone interfaces with carcinomatous invasion were distinguished. These included areas with bone resorption, smooth bone surface and new bone formation. In the bone-resorption area, numerous osteoclasts were located along the bone surface, which was surrounded by myxoid stroma. The myxoid stroma was characterized by immunopositivity for heparan sulfate proteoglycan (HSPG), abundant vascularity and macrophagic infiltration. In the bone-formation area, rows of osteoblasts were aligned on the bone surface. The stroma around osteoblasts was also HSPG-immunopositive, poor in vascularity but rich in activated fibroblasts. In the smooth-bone area, the stroma showed an organizing phase of granulation tissue with slender fibroblasts and mature collagen fibers but with less vascularity and inflammatory infiltrates. CONCLUSION: The results indicate that the stromal architecture, especially in terms of its inflammatory cellular, vascular and matrix compositions, is strictly regulated in the timing and site of jaw bone remodeling which is causes by carcinomatous invasion.
Antigens, CD↗
Maxillary odontogenic keratocyst with respiratory epithelium: a case report.
We report a case of odontogenic keratocyst with a respiratory epithelial lining and a malformed impacted tooth in the maxilla of a 39-year-old Japanese female who suffered from swelling symptoms for half a year. CT examinations revealed an air-filled cystic lesion with an impacted tooth crown in the maxillary bone which expanded to the nasal cavity as well as to the maxillary sinus. Histopathologically, the surgically removed cyst wall consisted of fibrous granulation tissue with a lining of parakeratinized squamous epithelium as well as ciliated pseudostratified epithelium and with retention of desquamated keratin materials in the lumen. The impacted tooth was malformed lacking a root portion. We discuss the frequency of respiratory epithelium in odontogenic keratocysts.
Adult↗
Melanoma mouse model implicates metabotropic glutamate signaling in melanocytic neoplasia.
To gain insight into melanoma pathogenesis, we characterized an insertional mouse mutant, TG3, that is predisposed to develop multiple melanomas. Physical mapping identified multiple tandem insertions of the transgene into intron 3 of Grm1 (encoding metabotropic glutamate receptor 1) with concomitant deletion of 70 kb of intronic sequence. To assess whether this insertional mutagenesis event results in alteration of transcriptional regulation, we analyzed Grm1 and two flanking genes for aberrant expression in melanomas from TG3 mice. We observed aberrant expression of only Grm1. Although we did not detect its expression in normal mouse melanocytes, Grm1 was ectopically expressed in the melanomas from TG3 mice. To confirm the involvement of Grm1 in melanocytic neoplasia, we created an additional transgenic line with Grm1 expression driven by the dopachrome tautomerase promoter. Similar to the original TG3, the Tg(Grm1)EPv line was susceptible to melanoma. In contrast to human melanoma, these transgenic mice had a generalized hyperproliferation of melanocytes with limited transformation to fully malignant metastasis. We detected expression of GRM1 in a number of human melanoma biopsies and cell lines but not in benign nevi and melanocytes. This study provides compelling evidence for the importance of metabotropic glutamate signaling in melanocytic neoplasia.
Animals↗