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Biomedical subjects

W G Zhang

Publications and source records attributed to W G Zhang.

At least 19 recordsLinked to original sources

Serological identification and bioinformatics analysis of immunogenic antigens in multiple myeloma.

Identifying appropriate tumor antigens is critical to the development of successful specific cancer immunotherapy. Serological analysis of tumor antigens by a recombinant cDNA expression library (SEREX) allows the systematic cloning of tumor antigens recognized by the spontaneous autoantibody repertoire of cancer patients. We applied SEREX to the cDNA expression library of cell line HMy2, which led to the isolation of six known characterized genes and 12 novel genes. Known genes, including ring finger protein 167, KLF10, TPT1, p02 protein, cDNA FLJ46859 fis, and DNMT1, were related to the development of different tumors. Bioinformatics was performed to predict 12 novel MMSA (multiple myeloma special antigen) genes. The prediction of tumor antigens provides potential targets for the immunotherapy of patients with multiple myeloma (MM) and help in the understanding of carcinogenesis. Crude lysate ELISA methodology indicated that the optical density value of MMSA-3 and MMSA-7 were significantly higher in MM patients than in healthy donors. Furthermore, SYBR Green real-time PCR showed that MMSA-1 presented with a high number of copy messages in MM. In summary, the antigens identified in this study may be potential candidates for diagnosis and targets for immunotherapy in MM.

Antigens, Neoplasm↗

Involvement of hypoxia-inducible factor-1-alpha in multidrug resistance induced by hypoxia in HepG2 cells.

Aim of the study was to explore the influence of hypoxia on multidrug resistance related genes and the potential role of hypoxia-inducible factor-1-alpha (HIF-1alpha) in formation of multidrug resistance in HepG2 human hepatocellular carcinoma cell line. HepG2 cells were subjected to hypoxia in a cohort of exposed time. A cell model stably expressing HIF-1alpha was established by liposome-mediated transfection of plasmid pcDNA3/HIF-1alpha into HepG2 cells. Apoptosis of HepG2 cells exposed to hypoxia or transfected by plasmid pcDNA3/HIF-1alpha was detected by Flow Cytometry after administration of chemotherapeutic drug (5-Fu). Real-time fluorescent quantitative PCR and Western-blot technique were used to analyze the expressions of multidrug resistance related genes mdr1, MRP1 and LRP at mRNA and protein level, respectively. Apoptosis Index of HepG2 cells exposed to hypoxia stepped down as exposed time extended after administration of 5-Fu. The expression of mdr1, MRP1 and LRP gene and protein revealed a hypoxic time-dependent induction and was synchronous with the alterations of HIF-1alpha in HepG2 cells exposed to hypoxia. The expressions of these multidrug resistance related genes were remarkably increased in HIF-1alpha transfected HepG2 cells as compared to empty vector transfected cells. Apoptosis index of HIF-1alpha transfected cells was obviously less than that of control cells when they were simultaneously exposed to 5-Fu for 24hrs. In conclusion, ambient hypoxia might be one of the causes for the formation of multidrug resistance in HepG2 human hepatocellular carcinoma cell line. Hypoxia-elicited multidrug resistance related protein expression might be a pathway for resistance of HepG2 cells to chemotherapeutics and HIF-1alpha might be involved in this process.

Apoptosis↗

The pre-styloid compartment of the parapharyngeal space: a three-dimensional digitized model based on the Chinese Visible Human.

To build a digitized visible model of the parapharyngeal space of the Chinese Visible Human and to provide a sectional anatomic basis for radiological and clinical diagnosis of the parapharyngeal space, sectional anatomy data of the parapharyngeal space were selected from the Chinese Visible Human male and female to compare with MR imaging findings in the axial planes. From these data the parapharyngeal space and surrounding structures were segmented. They were then reconstructed in three dimensions on PC. In the axial planes of the sectional anatomy and MR imaging, the shape, content and relations of the parapharyngeal space were clearly displayed and the dominant plane for showing the parapharyngeal space was elicited. The three-dimensional reconstructed images displayed perfectly the anatomic relationships of the parapharyngeal space, parotid, muscles, mandible and vessels. All reconstructed structures can be displayed singly, in groups or as a whole; any diameter or angle of the reconstructed structures can be easily measured. The Chinese Visible Human male and female data set can provide complete and accurate data. The digitized model of the parapharyngeal space and its surroundings offers unique insights into the complex anatomy of the area, providing morphologic data for imaging diagnosis and surgery of the parapharyngeal space.

Anatomy, Cross-Sectional↗

A study on the sectional anatomy of the oculomotor nerve and its related blood vessels with plastination and MRI.

To obtain normal images and sectional anatomical data of the oculomotor nerve and its related arteries, the optimal angles and the length of intracisternal segment of the oculomotor nerve were measured on MPR images. Meanwhile, the relationships between the nerve and the basilar, posterior cerebral, superior cerebellar and posterior communicating arteries were observed from plastination slices, original images, MPR and MIP images. MRI revealed similar results to the plastination sections. The intracisternal segment of oculomotor nerve formed an angle with the posterior plane of the brainstem. The angle was significantly smaller in individuals under 10 and over 50 years old ( P<0.05), and there was no marked difference in the angle between the oculomotor nerve and the median sagittal plane among the different groups ( P>0.05). Shift of the basilar artery was more likely to be found in aged individuals. Most of the posterior cerebral and superior cerebellar arteries were close to the nerve, and a few of them seemed to compress it; for the posterior communicating artery, only the embryonic type was close to or seemed to compress the nerve. MRI is an accurate imaging technique for determination of the relationship of the oculomotor nerve to its related arteries.

Adolescent↗

mTOR, a novel target in breast cancer: the effect of CCI-779, an mTOR inhibitor, in preclinical models of breast cancer.

The mammalian target of rapamycin (mTOR) is a central regulator of G1 cell cycle protein synthesis that precedes commitment to normal cellular replication. We have studied the effect of cell cycle inhibitor-779 (CCI-779), a rapamycin ester that inhibits mTOR function, on the proliferation of a panel of breast cancer cell lines. Six of eight lines studied were sensitive (IC(50)< or = 50 nM) and two lines were resistant (IC(50)>1.0 microM) to CCI-779. Sensitive lines were estrogen dependent (MCF-7, BT-474, T-47D), or lacked expression of the tumor suppressor PTEN (MDA-MB-468, BT-549), and/or overexpressed the Her-2/neu oncogene (SKBR-3, BT-474). Resistant lines (MDA-MB-435, MDA-MB-231) shared none of these properties. CCI-779 (50 nM) inhibited mTOR function in both a sensitive and a resistant line. In nu/nu mouse xenografts, CCI-779 inhibited growth of MDA-MB-468 (sensitive) but not MDA-MB-435 resistant tumors. Treatment of sensitive lines with CCI-779 resulted in a decrease in D-type cyclin and c-myc levels and an increase in p27(kip-1) levels. There was good correlation between activation of the Akt pathway and sensitivity to CCI-779. Amplification of mTOR-regulated p70S6 kinase, which is downstream of Akt, may also have conferred CCI-779 sensitivity to MCF-7 cells. Taken together, the data suggest that mTOR may be a good target for breast cancer therapy, especially in tumors with Akt activation resulting from either growth factor dependency or loss of PTEN function.

Animals↗

Effect of Salvia miltiorrhiza Bunge injection on anticardiolipin antibody production induced by beta2 glycoprotein.

AIM: To explore the therapeutic effect and the mechanism of Chinese herbs on antiphospholipid syndrome (APS) by observing the effect of Salvia miltiorrhiza Bunge injectio (SmBI) on anticardiolipin antibody (aCL) induced by beta2 glycoprotein I (beta2-GP I). METHODS: Sixty female mice randomly fell into 6 groups: group A, B, C, D was injected through abdominal cavity with different dosage of SmBI daily; after 14 d, group A, B, C, E was immunized with 150 microg of purified human beta2-GP I in complete Freund's adjuvant subcutaneously; group F as control. The titre of aCL were detected by enzyme linked immunosorbent assay; subsets of T cell were grouped by streptavidin-biotin complex technique; and the activity of IL-2 was measured by MTT chromatometry. RESULTS: (1) Compared with group E, the absorbance (A) of aCL in group A, B, and C was decreased (P < 0.05 or P < 0.01). By linear correlation, the dosage is negatively correlated with the A values of aCL in 1, 2, and 3 weeks (P < 0.01). (2) Compared with group E, TH/TS ratio was reduced in group A, B, and C (P < 0.05 or P < 0.01); there is no significant differences between group D and F (P>0.05). By linear correlation, the dosage is negatively correlated with TH/TS ratio (P < 0.01). (3) Compared with E, the activity of IL-2 in group B and C decreased significantly (P < 0.01). By linear correlation, there is negative correlation between dosage and IL-2 activity (P < 0.01). There is no significant difference between D and F (P > 0.05). (4) There is positive correlation between TH/TS ratio and IL-2 activity in different dilutions (P<0.01). CONCLUSION: The mechanism of suppressive effect of SmBI on aCL induced by beta2-GP I may be realized by resuming the elevated TH/TS ratio and IL-2 activity. The state that SmBI have no effect on normal mice indicates that SmBI has selective immunoregulative functive.

Adjuvants, Immunologic↗

[A study of fluoride and dexamethasone affecting the expression of growth factors in osteoblast].

OBJECTIVES: Investigate the expression of growth factors or receptors in osteoblast, to understand the possible mechanism of low dose fluoride and dexamethasone on the development and differentiation of osteoblast. METHODS: Investigate the expression of platelet derived growth factors (PDGF-A) and insulin-like growth factors (IGF-I-Ralpha) in osteoblast cultured in vitro by immunohistochemistry, furthermore, to deal with the results by image analysis. RESULTS: There were no significant effects of PDGF-A both of low doses fluoride and dexamethasone treated on the MC3T3-E1 osteoblast during the various stages of the development. Otherwise, the expression of IGF-I-Ralpha might increase treated with them at the early stage of the cell culture, and dexamethasone did more significantly. CONCLUSION: Low doses of fluoride or dexamethasone may promote the expression of IGF-I-Ralpha in osteoblast precursors, which maybe contribute to the effects of fluoride and dexamethasone on the proliferation and differentiation of osteoblast.

English Abstract↗

[Construction, expression and activity test of a reshaping single-chain antibody against human CD3].

Monoclonal antibody (McAb) against human CD3 can adjust human body's immune statement in various ways, so that its clinical potential is highly regarded. In order to overcome the immunogenecity related to the murine McAb, this research effort was focused on constructing a reshaping single-chain antibody(scFv) against human CD3 employing antibody engineering. First, the CDRs of the murine McAb against human CD3 OKT3 was transplanted into the light-chain framework regions (FRs) of human McAb LS1 and the heavy-chain FRs of human McAb Nd respectively, spatial conformation was predicted by computer analysis. Then some particular residues were replaced in FRs basing on the result of conformational prediction to draw out the amino acid sequences of the reshaped VL and VH. The genes were chemically synthesize and inserted into an expression vector pROH80 to construct the reshaping scFv. Inducing the expression of reshaping scFv, the products are mainly as inclusion bodies. The reshaping scFv was expressed in another vector pALM. The inclusion bodies were denatured and then renatured by gel filtration. The renatured products were purified by immobilized metal affinity chromatograph (IMAC). Finally, the antigen-binding activily of the reshaping scFv against human CD3 was testified by the Compelitire in hibilory fluorescenceactivated cell sorting (FACS). The competitive inhibition rate is 18%.

Amino Acid Sequence↗

The expression of E-cadherin-catenin complex in adenoid cystic carcinoma of salivary glands.

OBJECTIVE: The aim of this study was to investigate the expression of the E-cadherin-catenin (E-cad-cat) complex in human salivary adenoid cystic carcinoma (ACC) and determine the effects of E-cad-cat expression on tumor differentiation, metastasis, and invasion, as well as its clinical significance. METHODS: The expression of adhesion molecules, which included E-cadherin, alpha-catenin, and beta-catenin, on 10 cases of normal salivary glands, and 50 cases of human salivary ACC were investigated by immunohistochemistry. RESULTS: Strong and stable expression of E-cad-cat complex showed in normal salivary glands tissues. However, weaker expression and even complete absence, of the positive signals were shown on all ACC cases. Coexpression of E-cad-cat were detected in 29 (58%) of the 50 ACC specimens. Positive staining of E-cad-cat complex were detected in 7 (39%) of the 18 ACC primary sites that had aggressive phenotype of nerve and vasculature. CONCLUSION: Strong positive signals of E-cad-cat complex are shown in normal salivary glands, and the down-regulated expression were observed in ACC cases. The expression of E-cad-cat complex is associated with a more aggressive phenotype on nerve and vasculature. No association with various histopathologic subtypes was found. However, absent or low E-cad expression was observed more frequently in solid carcinomas than in cribriform or tubular carcinomas. Down-regulation of E-cad-cat complex may be associated with canceration of normal salivary glands. The expression levels of E-cad-cat were positively correlated to the differentiation of ACC tumor cells.

Antibodies, Monoclonal↗

[A study of fluoride and dexamethasone affecting the proliferation in osteoblast].

OBJECTIVE: Investigate the effects of low dose of fluoride and dexamethasone on the proliferation of MC3T3-E1 osteoblast cell line in vitro, which was for determining the possible effects of fluoride and dexamthasone affecting the development and differentiation of osteoblast. METHODS: Observe the growing shape of MC3T3-E1 osteoblast cultured in vitro, and investigate the effects of low dose of fluoride and dexamethasone on the proliferation of osteoblast by cell count. RESULTS: MC3T3-E1 osteoblast grew stably. The cells in D group increased obviously in the early stage of the cell culture, however the cells in F group had no tendency of increasing during the whole culture. CONCLUSION: In vitro low doses of dexamethasone may promote the proloferation of osteoblast precursors,however,low dose of fluoride has no effect on promoting proliferation of MC3T3-E1 osteoblast.

English Abstract↗

Wild-type p53 protein potentiates phototoxicity of 2-BA-2-DMHA in HT29 cells expressing endogenous mutant p53.

To better understand the effects of p53 on the process of photodynamic therapy (PDT)-induced cell death, we introduced a wild-type p53 gene into the HT29 colorectal carcinoma cell line, which bears an endogenous mutant p53, using a lipofectin system. The influence of p53 status on the sensitivity induced by 2-butylamino-2-demethoxy-hypocrellin A (2-BA-2-DMHA) photosensitization was then examined. The results indicate that infection with wild-type p53 induces a growth arrest but does not induce cell death, and sensitizes the cells to PDT. At a concentration of 5 microM 2-BA-2-DMHA with a red light of 18 J/cm2 (lambda = 600-700 nm), the survival is reduced from 58.72% in HT29 cells to 13.49% in wild-type p53-infected HT29 cells. Apoptosis following PDT appears earlier in HT29 cells infected with wild-type p53 than in parent HT29 cells and empty vector-infected HT29 cells. These findings suggest that although wild-type p53 is, by itself, insufficient to induce apoptosis in cells with p53 mutation, it enhances the photosensitivity of 2-BA-2-DMHA by strongly potentiating the induction of apoptosis.

Apoptosis↗

Antisense bcl-2 retrovirus vector increases the sensitivity of a human gastric adenocarcinoma cell line to photodynamic therapy.

The bcl-2 oncoprotein directly prolongs cellular survival by blocking apoptosis and its overexpression is associated with cellular resistance to killing by chemotherapeutic drugs and gamma-irradiation. Meanwhile, it has been shown that bcl-2 antisense oligonucleotide can induce apoptosis or increase toxicity of the treatment in tumors in vivo and in vitro. However, it is difficult to obtain stable transfection by this approach and there are no reports about the effect of an antisense bcl-2 on the sensitivity to oxidative stress induced by photodynamic therapy (PDT). Here we investigated the effect of an antisense bcl-2 RNA retrovirus vector transfer on the sensitivity of 2-butylamino-2-demethoxy-hypocrellin A (2-BA-2-DMHA) photosensitization in a human gastric adenocarcinoma MGC803 cell line. The results indicate that antisense bcl-2-infected MGC803 cells expressed exogenous antisense bcl-2 mRNA measured by reverse transcription polymerase chain reaction and significantly reduced bcl-2 protein determined by western blotting analysis. The decreased expression of bcl-2 protein was accompanied by increased phototoxicity and susceptibility to apoptosis induced by 2-BA-2-DMHA PDT. Our finding suggests that reduction of bcl-2 protein in gastric cancers, and possibly also in a variety of other tumors, may be a novel and rational approach to improve photosensitivity and the treatment outcome.

Adenocarcinoma↗

[Construction of expressing vector for phage display scFv and a mouse unspecified antibody library].

The display of antibody gene library on the surface of E. coli filamentous phage offers a new method of obtaining antibodies against specific antigens. In the paper, it is reported that a phagemid for phage display antibody, named after pFUW80, characterized expressing scFv either secretly or stickily. With a series of designed PCR primers, heavy-chain and light-chain variable region genes of mice antibodies were amplified and a mouse unspecified single-chain antibody library with the size of 1.2 x 10(6) clones was constructed. From this library, phage was selected out against antigen human IgG and detected by ELISA and partial sequence analysis. These primary results established the foundation for future research using this system.

Animals↗

In vitro study of the effects of endothelin-1 on human dental pulp cells.

OBJECTIVE: To study the effect of Endothelin-1 (ET-1) on cell proliferation in primary cultures of human dental pulp cells; the cell mechanism of mitogen; and collagen synthesis. METHODS: Primary cultures of human dental pulp cells; Thiazolyl Blue Tetrazolium Bromide (MTT) test to investigate cell proliferation; measurement of calcium ions in human dental pulp cells with the calcium-sensitive dye fluo-3; enzyme-linked immunosorbent assay and avidin-biotin complex immunocytochemical assay for collagen synthesis in human dental pulp cells. RESULTS: Incubation of cultures with ET-1 resulted in a dose-dependent increase in cell proliferation over control (unstimulated) cultures. In addition ET-1 increased the Ca2+ concentration in human dental pulp cells, which cannot be inhabited by Verapamil. The finding demonstrated that collagen type I synthesis of human dental pulp cells was increased by ET-1. CONCLUSIONS: ET-1 played a crucial role in pulp restoration of lesions and had an effect on mitogen and differentiation of human dental pulp cells.

Cell Division↗

[An in vitro study of anti acidic etching effect of polypeptide-containing toothpaste on dental enamel]

OBJECTIVE:The present study was to investigate the anti-acidic etching effect of a kind of toothpaste containing biotin-polypeptide. METHODS:28 enamel slabs were treated with polypeptide-containing toothpaste in the pH-cycle environment,and then were analyzed by micro-hardness detector and polarized light microscope.RESULTS:The dental enamel treated with polypeptide-containing toothpaste was harder than the control group (P<0.0001);and the degree of demineralization in treated group was also less than the control one.CONCLUSION:There was certain acid resistant effect using the polypeptide containing toothpaste to treat the dental enamel.

Journal Article↗

[The expression of CD44s and CD44v6 in human salivary adenoid cystic carcinoma ]

OBJECTIVE:The present study was to investigate the expression of CD44s and CD44v6 in human salivary adenoid cystic carcinoma (ACC) and to discuss the relationship between the expression of the proteins and the biological characteristic of ACC.METHODS:Formalin fixed,paraffin embedded sections in 10 cases of normal salivary glands and 50 cases of human salivary ACC were examined using immunostaining for CD44s and CD44v6.RESULTS:(1)Positive signals of CD44s and CD44v6 were detected in all the specimens of normal salivary glands.Positive signals of CD44s and CD44v6 were detected in 29(58%) and 35(70%) of the 50 cases of ACC.(2)The expression of CD44s and CD44v6 in the solid pattern of ACC was significantly less than that in the cribriform and tubular pattern of ACC(P<0.05).(3)Positive CD44s staining was detected in 3(13.6%) of the 22 ACC primary sites which had a local recurrence after operation.CONCLUSION:(1)Adhesion molecules including CD44s and CD44v6 may contribute to maintain the normal structures in salivary gland tissues.(2)T he down-regulation or loss of CD44s expression was contributed to the recurrence of human salivary ACC and CD44s could be regarded as one of the factors to evaluate the prognosis of ACC patients.

Journal Article↗