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Biomedical subjects

Min Zheng

Publications and source records attributed to Min Zheng.

At least 55 records · Page 3Linked to original sources
PubMed2004

Modification of the brain-derived neurotrophic factor gene: a portal to transform mesenchymal stem cells into advantageous engineering cells for neuroregeneration and neuroprotection.

Multipotential mesenchymal stem cells (MSCs) are ideal seed cells for recruiting the loss of neural cells due to their strong proliferative capacity, easy acquisition, and considerable tolerance of genetic modifications. After transduction of brain-derived neurotrophic factor (BDNF) gene via recombinant retroviral vectors into the human MSCs, nearly 100% of cells expressed BDNF (which were therefore transformed into BNDF-MSCs) as detected by immunocytochemistry, and the quantity of BDNF in the culture medium was increased by approximately 20,000-fold. In spite of the genomic integration of an exogenous gene, BDNF-MSCs did not present any structural aberration in the chromosomes. All-trans-retinoic acid (RA) induction caused the BDNF-MSCs to differentiate into neural cells with significantly increased expressions of such neural-specific proteins as nestin, NeuN, O4, and glial fibrillary acidic protein (GFAP). The voltage-dependent K+/Ca2+ currents were recorded from the induced BDNF-MSCs using patch-clamp technique. Compared with the MSCs induced by both RA and BDNF, BDNF-MSCs survived in significantly greater number in the induction medium, and also more cells were induced into neuron-like cells (NeuN, P < 0.01) and oligodendrocyte-like cells (O4, P < 0.05). We suppose that, once engrafted into human central nervous system, the BDNF-MSCs would not only recruit the neuronal losses, but also provide, by way of paracrine, large quantities of BDNF that effectively perform the functions of neuroprotection and neuroregeneration, promoting the activation of endogenous neural stem/progenitor cells and their chemotactic migration. On the other hand, the BDNF-MSCs that can survive in the host environment and differentiate subsequently into functional mature cells may also serve as specifically targeting vectors for ex vivo gene therapy.

Adult↗
PubMed2004

TRIO amplification and abundant mRNA expression is associated with invasive tumor growth and rapid tumor cell proliferation in urinary bladder cancer.

Studies by comparative genome hybridization have suggested that 5p amplification is related to tumor progression in urinary bladder cancer. In this study seven genes (TAS2R, ADCY2, DNAH5, CTNND2, TRIO, ANKH, and MYO10) located to 5p15.31-5p15.1 were analyzed by fluorescence in situ hybridization using a tissue microarray containing samples from tumors and cell lines with known 5p amplification by comparative genome hybridization. Amplification frequency was highest for TRIO, which maps to 5p15.2 and encodes a protein with a putative role in cell-cycle regulation. To further investigate the role of TRIO amplification in bladder cancer, a tissue microarray containing samples from 2317 bladder tumors was used for fluorescence in situ hybridization analysis. TRIO amplification was strongly associated with invasive tumor phenotype, high tumor grade, and rapid tumor cell proliferation (Ki67 LI) (P < 0.0001 each). Only 7 of 456 pTaG1/G2 tumors (1.5%) but 62 of 485 pT1-4 carcinomas (12.8%) had TRIO amplification. TRIO amplification was not associated with poor prognosis. Using a frozen bladder tumor tissue microarray RNA in situ hybridization confirmed that TRIO is up-regulated in amplified tumors. It is concluded that TRIO up-regulation through amplification has a potential role in bladder cancer progression.

Carcinoma↗
PubMed2004

An epididymis-specific beta-defensin is important for the initiation of sperm maturation.

Although the role of the epididymis, a male accessory sex organ, in sperm maturation has been established for nearly four decades, the maturation process itself has not been linked to a specific molecule of epididymal origin. Here we show that Bin1b, a rat epididymis-specific beta-defensin with antimicrobial activity, can bind to the sperm head in different regions of the epididymis with varied binding patterns. In addition, Bin1b-expressing cells, either of epididymal origin or from a Bin1b-transfected cell line, can induce progressive sperm motility in immotile immature sperm. This induction of motility is mediated by the Bin1b-induced uptake of Ca(2+), a mechanism that has a less prominent role in maintaining motility in mature sperm. In vivo antisense experiments show that suppressed expression of Bin1b results in reduced binding of Bin1b to caput sperm and in considerable attenuation of sperm motility and progressive movement. Thus, beta-defensin is important for the acquisition of sperm motility and the initiation of sperm maturation.

Animals↗
PubMed2004

Variable expression of human lactoferrin gene in mice milk driven by its 90 KB upstream flanking sequences.

One major drawback in research of animal mammary gland bioreactors is the low production rate of high-expressing transgenic animals due to position effects. To obtain high and stable expression of foreign gene, yeast and bacterial artificial chromosome have been used as transgene vector in recent research. Human lactoferrin is a bioactive, versatile protein, and has large potential in nutritional and therapeutic applications. Therefore, production of recombinant lactoferrin using animal bioreactors was studied widely to satisfy its large requirement. We reported here a transgenic mice model with high-level expression of recombinant human lactoferrin in mammary gland. Transgene construct used here was a human bacterial artificial chromosome containing intact lactoferrin-encoding transcript unit, approximately 90 kb 5'-flanking sequences and 27.2 kb 3'-flanking sequences. We obtained totally 10 transgenic mice whereas two of them lacked of part of upstream sequences of the gene. Milk of eight transgenic mice line was detected by Western blot and radioimmunoassay and seven lines expressed recombinant human lactoferrin at high but variable level (0.29, 0.53, 0.90, 1.23, 2.76, 3.58, and 8.02 mg/mL, respectively). The variability of expression indicates that even the 90 kb 5' flanking sequence of the transgene can't overcome position effects completely. Moreover, we also determined sequences of 9.3 kb regulatory region and 10.6 kb encoding region of the gene and thus supplemented all unknown sequences. Our results suggested that transgene vector used here has potential to be used in large farm animals for production of recombinant human lactoferrin in industrial scale.

Animals↗
PubMed2004

[Skin-like structure generated from implantation of hair follicle bulb cells into collagen/chitosan porous scaffolds in vitro].

OBJECTIVE: To observe the skin regeneration after hair follicle bulb cells were implanted into collagen/chitosan porous scaffolds in vitro. METHODS: The cultured dorsal hair follicle bulb cells of 4d-old C57BL/6J mice were implanted into collagen/chitosan porous scaffolds in vitro. The skin regeneration was observed. RESULT: The skin-like structure was formed on the collagen/chitosan porous scaffolds where were cultured the hair follicle bulb cells before 4th passages. CONCLUSION: The skin-like structure is generated in vitro when early passages of cultured hair bulb cells are implanted into collagen/chitosan porous scaffolds.

Animals↗
PubMed2004

[Hair follicle regeneration after implantation of hair follicle cells into subcutis of nude mice].

OBJECTIVE: To observe the hair follicle regeneration after implantation of hair follicle cells into the subcutis of nude mice. METHODS: The cultured hair papilla cells,dermal sheath cells and fibroblast of human scalp were mixed with the cells of hair follicle epithelium in different ratio, and then implanted into the subcutis of nude mice. The regeneration of hair follicle was observed. RESULT: The hair follicle-like structure was formed in cluster where the cultured hair follicle epithelium cells were mixed with hair papilla cells. But no hair follicles were formed where the hair follicle epithelium was implanted with dermal sheath cells or fibroblasts. CONCLUSION: The hair follicle-like structure is generated in vivo when the mixed cells of early passages cultured hair papilla cells with hair follicle epithelium are implanted into the subcutis of nude mice.

Animals↗
PubMed2004

[Protective effects of minocycline against hair follicle damage induced by cytosine arabinoside in vitro].

OBJECTIVE: To investigate the protective effects of minocycline against hair follicle damage induced by cytosine arabinoside (Ara-c). METHODS: An in vitro organ culture of mouse vibrissa follicles was used and different concentrations of Ara-c and minocycline were added in the culture media. The total growth length, growth speed and growth period of hair were observed with invert microscopy and the survival of hair bulb cells was measured by MTT method. RESULT: Minocycline (0.3 x 10(-6) approximately 10(-5) mol/L) improved hair follicle total growth length, growth speed and hair growth period and also improved survival of hair bulb cells in vitro organ culture, which were inhibited by Ara-c. CONCLUSION: Minocycline can protect hair follicle directly from damage induced by Ara-c.

Animals↗
PubMed2004

[Expressions of bFGF, ET-1 and SCF in dermal papilla cells and the relation to their biological properties].

OBJECTIVE: To investigate the expression of bFGF, ET-1 and SCF in different passages of cultured dermal papilla cells (DPC), and their possible effect on biological behaviour of DPC. METHODS: The expression of bFGF, ET-1 and SCF in different passages of cultured DPC was detected by immunocytochemistry and in situ hybridization. RESULT: The expression of ET-1 and SCF in early passages of cultured DPC was stronger, but became negative in late passages (>6 passages). The stronger the expression of ET-1 and SCF in DPC, the higher ability of DPC to induce hair follicle regeneration. CONCLUSION: The expression strength of ET-1 and SCF is related to the ability of DPC inducing hair follicle regeneration.

Endothelin-1↗
PubMed2004

[Effect of STAT3 phosphorylation and p53 expression on human epidermal non melanoma cutaneous tumors].

OBJECTIVE: To investigate the effect of stat3 phosphorylation and p53 expression on human epidermal non-melanoma cutaneous tumours. METHODS: Immunohistochemistry technique was employed to measure the expression of p-stat3 and p53 protein in skin tissue from 30 cases of skin squamous cell carcinoma (SCC), 20 cases of basal cell carcinoma (BCC), 20 cases of seborrhoeic keratosis (SK) and 20 normal subjects. RESULT: (1) p-stat3 protein was abnormally increased in SCC and BCC as compared with normal skin and SK. Expression of p-stat3 in SCC was also significantly higher than that in BCC. (2) Expression of p-stat3 was higher in poorly-differentiated cancers than that in well-differentiated cancers in SCC. The positive rate of p-stat3 expression was correlated with the depth of tumor invasion, but not with tumor size. (3) There was no p53 protein expression on normal skin and SK, it was significantly upregulated in SCC and BCC. In SCC, the intensity of p53 expression was associated with tumor differentiation. There was no correlation between the positive rate of p53 expression and the depth of tumor invasion, whereas the positive rate of p53 expression was correlated with the sun-exposure area. (4) There existed positive correlation between the expression intensity of p-stat3 and p53 in SCC (r=0.641, P<0.05). CONCLUSION: (1) The overexpression of p-stat3 may play an important role in the development of epidermal tumors. (2) The abnormal activation of stat3 may be related to metastatic potentials in SCC. (3) Both p53 gene and stat3 may contribute to the pathogenesis of skin SCC.

Basal Cell Carcinoma↗
PubMed2004

[Relationship of serum levels of HGF and MMP-9 with disease activity of patients with systemic lupus erythematosus].

OBJECTIVE: To determine the relationship of serum hepatocyte growth factor (HGF) and matrix metalloproteinase-9 (MMP-9) levels with the disease activity of systemic lupus erythematosus (SLE). METHODS: Serum levels of HGF and MMP-9 were measured by ELISA in 36 patients with SLE and 30 healthy subjects as controls. RESULT: (1)Significantly increased serum level of HGF was found in SLE patients as compared with that in healthy controls (P<0.001), but serum level of MMP-9 in SLE patients decreased (P<0.001). Serum level of HGF was significantly decreased after treatment in SLE patients (P<0.05), but serum level of MMP-9 was increased (P<0.05). (2)Serum level of HGF was markedly higher in patients with active disease (24 cases) than those with inactive disease (P<0.05), but serum level of MMP-9 was lower (P<0.05). (3)Significantly increased serum level of HGF was found in patients with renal damage (16 cases) than those without (P<0.001), but serum level of MMP-9 was lower in patients with renal damage than those without (P<0.005). (4)Serum level of HGF was higher in patients with arthritis (23 cases) than those without (P<0.01), but serum level of MMP-9 had no significant difference in two groups (P>0.05). (5)The area of ROC curve was 0.707 and the sensitivity was 66.7% if serum level of HGF was served as diagnostic standard. The area of ROC curve was 0.984 and the sensitivity was 97.2% if serum level of MMP-9 was served as diagnostic standard. The sensitivity was 66.7% (24/36) if the two markers were examined simultaneously. CONCLUSION: The data suggest that HGF and MMP-9 could be involved in the pathogenesis of SLE, and serum levels of HGF and MMP-9 be used as markers to monitor disease activity, renal damage, disease progression and amelioration in SLE.

Adolescent↗
PubMed2004

[Suppression of Tacrolimus on the expression of MMP-9 in rhHGF-stimulated ECV304 cell line].

OBJECTIVE: To investigate the stimulating effect of recombinant human hepatocyte growth factor (rhHGF) on the expression of matrix metalloproteinase-9 (MMP-9) in ECV304 cells and the suppression effect of FK506. METHODS: The growth curve of ECV304 cells was drawn and the valid concentration of rhHGF and FK506 were determined. The expression levels of MMP-9 were analyzed by flow cytometry (FCM). RESULTS: (1) The optimal growth concentration of ECV304 cell was 1.0E+4/ml and logarithm growth time was 3-5 days. The valid concentration of rhHGF was 8 ng/ml and IC50 was 8.27 ng/ml. The valid concentration of FK506 was 50 ng/ml and IC50 was 51.63 ng/ml. (2) The expression level of MMP-9 was 39.74% in controls. Cell livability was 1.388169 +/-0.102033 (P<0.01) when 8 ng/ml rhHGF was added to culture medium, and the expression level of MMP-9 was 40.32%. Cell livability was 0.398764 +/-0.092476 (P<0.01) when 50 ng/ml FK506 was added to culture medium, and the expression level of MMP-9 was 14.61%. Cell livability was 0.767203 +/-0.02639 (P<0.01) when 8 ng/ml rhHGF was added 15 minutes after 50 ng/ml FK506 was added to culture medium, and the expression level of MMP-9 was 35.08%. CONCLUSION: rhHGF can stimulate the proliferation of ECV304 cells and increase the protein expression level of MMP-9. FK506 can suppress the proliferation of ECV304 cells and decrease the expression level of MMP-9. FK506 can inhibit the proliferation of ECV304 cells induced by rhHGF and decrease MMP-9 protein level.

Cell Division↗
PubMed2004

[Transplantation of exogenous mesenchymal stem cells for treatment of myocardial infarction in rat].

OBJECTIVE: To investigate the possibility of homing,survival, migration and differentiation of rat mesenchymal stem cells (rMSCs) in host myocardium and the effects of rMSCs transplantation on the function of heart with myocardial infarction; To observe the feasibility of using exogenous adult stem cells for cell therapy. METHODS: 35 female rats were separated randomly into three groups: normal control group (control, n = 10), acute myocardial infarction control group (AMI, n = 10) and myocardial infarction plus cell transplantation group (AMI + Cell, n = 15). The infarcted hearts were made by occlusion of left coronary artery. Male Wistar rats MSCs were isolated and purified. Then the cells were implanted into the infarcted hearts of female Wistar rats. Heart functions were measured 10 weeks after implantation. The hearts were harvested 10 weeks after implantation for immunohischemistry. RESULTS: (1) Purified rMSCs survived and homed in exogenous host hearts; (2) DAPI-labelled rMSCs with oval nucleus were widely distributed and stained positively of cardiac specific proteins. The arrangements of the donated cells paralleled with host myocardium fibers in infarcted host hearts. Transplantation of rMSCs was associated with a significant decrease of end left ventricular volumes, increase of left ventricular end-systolic pressure and increase of ratio of left ventricular pressure rise and left ventricular pressure decay (+/- dp/dt) (P < 0.05) as compared with the control hearts. The numbers of blood vessels were increased at the boundary of infarction site and the sizes of infarction were decreased significantly (P < 0.05). CONCLUSIONS: The purified rMSCs can survive in exogenous host hearts without addition of any immunosuppressant. These exogenous cells take part in myocardium regeneration and have beneficial effects on heart function.

Animals↗
PubMed2004

[The diagnostic value of eight serum indices for liver fibrosis].

OBJECTIVES: Using receiver operating characteristic (ROC) curve to compare the value of platelet derived growth factor-BB (PDGF-BB), transforming growth factor-beta1 (TGF-beta1), matrix metalloproteinase-1 (MMP-1), tissue inhibitor of matrix metalloproteinase-1 (TIMP-1), hyaluronic acid (HA), type III procollagen (PCIII), type IVcollagen (C-IV) and laminin (LN) in serum and message RNA (mRNA) expression of TIMP-1 and MMP-1 in peripheral blood mononucleocytes (PBMC) to diagnose liver fibrosis. METHODS: Serum samples from 60 chronic hepatitis B patients and 20 healthy blood donors were assayed for serum level of PDGF-BB, TGF-beta1, MMP-1 and TIMP-1 with ELISA, and for serum level of HA, PCIII, C-IV and LN, with RIA. The mRNA expression of TIMP-1 and MMP-1 in PBMC was detected by RT-PCR. Liver biopsy was performed in all those patients. The biopsy materials were examined histopathologically. RESULTS: Through the analysis by ROC curve, serum PDGF-BB is the most valuable marker, and its sensitivity was the highest among eight indices. The marker with the highest specificity was TIMP-1 mRNA in PBMCs. The area under the curve (AUC) of PDGF-BB, TIMP-1, HA, PCIII, C-IV, LN, TIMP-1 mRNA was 0.985, 0.726, 0.318, 0.728, 0.727, 0.583, 0.463, 0.876, respectively. The sensitivity and specificity of combination of PDGF-BB and TIMP-1 mRNA were 97.4%, 95.0%, respectively. CONCLUSION: Serum PDGF-BB is the most potential index among eight markers. The combination of serum PDGF-BB and TIMP-1 mRNA in PBMC might be more efficient to screen liver fibrosis.

Adult↗
PubMed2004

[Analysis of gene expression profiles among 3 epithelial ovarian tumor subtypes using cDNA and tissue microarrays].

BACKGROUND & OBJECTIVE: Ovarian cancer is the leading cause of death among the gynecological malignancy mainly due to lacking of effective early diagnostic methods. To identify novel candidate genes and further explore their clinical significance of epithelial ovarian tumors, we developed a new method in our laboratory by combining cDNA microarray with RNA in situ hybridization in frozen tissue microarray. METHODS: cDNA microarrays were used to seek differentially expressed genes among 3 subtypes of ovarian tumors (serous borderline ovarian tumors, serous ovarian cancers, and endometrioid ovarian carcinomas). RNA in situ hybridization in frozen tissue microarray was used to further confirm the findings from cDNA microarrays. RESULTS: In the study of cDNA microarray, 40 genes and ESTs showed significant differential expression between low and high-malignancy, as well as serous and endometrioid carcinomas. EPHB6, PTPRF, GFER, ERG25, PLRP1, FLJ22060, and WISP2 were further validated by RNA in situ hybridization in tissue microarray. CONCLUSIONS: cDNA microarray combined with RNA in situ hybridization in frozen tissue microarray is an ideal choice for identifying novel oncogenes. EPHB6, PTPRF, GFER, ERG25, PLRP1, FLJ22060, and WISP2 might become the new candidate oncogenes for epithelial ovarian cancer.

CCN Intercellular Signaling Proteins↗
PubMed2004

Chrysanthemum morifolium attenuated the reduction of contraction of isolated rat heart and cardiomyocytes induced by ischemia/reperfusion.

The present study was aimed to investigate the effect of Chrysanthemum morifolium Ramat. (CM) on isolated rat heart and ventricular myocytes during ischemia/anoxia and reperfusion/reoxygenation. The ischemia/reperfusion injury was induced by ligation the left artery descending coronary of isolated rat heart for 30 min followed by 30 min reperfusion with Langendorff equipment. Cell contraction in enzymatically isolated ventricular myocytes was determined by a video tracking system. The results showed CM (0.25 g/L to 1.0 g/L) increased left ventricular developed pressure (LVDP), +/- dp/dt(max), LVDP x HR and coronary flow (CF) and decreased heart rate (HR) in dose dependent manner. CM (0.5 g/L) attenuated the reduction of LVDP, +/- dp/dt(max) and CF caused by ischemia/reperfusion. CM (0.25 g/L to 1.0 g/L) increased peak velocity of cell shortening/relengthening (+/- dL/dt(max)) and contraction amplitude (dL) of isolated ventricular myocytes in a dose-dependent way under control condition, but without significant effect on end-diastolic cell length (L0). Under anoxia 5 min followed by 10 min reoxygenation, CM attenuated the reduction in contractile parameters. The results suggest that CM processes cardioprotective effect during ischemia/anoxia and reperfusion/reoxygenation in the isolated rat heart and the ventricular myocytes.

Animals↗
PubMed2004

[Isolation of genomic DNA from Cinnamomum cassia Presl].

OBJECTIVE: To extract high-quality DNA for RAPD and other molecular biology research from Chinese traditional medicine Cinnamomum cassia Presl.. METHOD: Improved CTAB method, low pH medium with high salt method (LPHS) and urea method were used for DNA extract. The yield and quality of DNA was analysed by electrophoresis, ratios of A260/A280 and PCR amplification. RESULTS: The modified CTAB method and low pH medium with high salt method can produce high qualiy DNA for PCR amplification.

Cetrimonium Compounds↗