Fuzheng Huayu recipe promotes the fenestration of capillarization in liver sinusoidal endothelial cells / 中南大学学报(医学版)

OBJECTIVE@#To explore the effect of Fuzheng Huayu (FZHY) recipe on the fenestration of capillarization in liver sinusoidal endothelial cells (LSECs).
@*METHODS@#Ten Sprague Dawley (SD) rats were fed with 0.46 g/kg FZHY powder by intragastric administration. Two hours later, a second gavage were given to the rats. The serum from rat heart at 1 hour after second gavage was collected (FZHY group, n=10). Another ten SD rats was administrated with distilled water through the same process and served as the control (control group, n=10). The serum from both groups were separately diluted with Dulbecco minimum essential medium (DMEM) for 10% and served as the culture medium for LSECs. At the different conditions, the vWF and CD31 expressions were detected by immunocytochemistry and Western blot, while the changes of LSECs fenestrae structure were observed under scanning electron microscopy.
@*RESULTS@#1) Immunocytochemistry and Western blot showed that the vWF and CD31 protein levels were lower in LSECs in the FZHY group than those in the control group. The gray levels of vWF and CD31 protein were 0.548±0.020 and 0.262±0.010 in the FZHY group, and 0.845±0.090 and 0.383±0.010 in the control group respectively, with statistical significant difference (t=5.18, 9.61, both P<0.05). 2) The results from scanning electron microscopy showed that the fenestration of LSECs was closed and almost lost in the control group, but many fenestra appeared in the LSECs in the FZHY group.
@*CONCLUSION@#FZHY recipe can suppress the expression of vWF and CD31, increase the fenestrae on the LSECs surface and reverse the capillarization of LSECs.

Identification of aberrantly expressed miRNAs in rectal cancer / 中南大学学报(医学版)

OBJECTIVE@#To identify aberrantly expressed miRNAs in rectal cancer.@*METHODS@#We used the miRCURY™ Array® LNA microRNA chip (v.14.0) to evaluate miRNA expression levels between rectal cancer tissues and adjacent non-tumor tissues; an average change more than 2-fold (and P value less than 0.05) was set as a cutoff level. All 6 paired rectal cancers were classified pathology stage C or D.@*RESULTS@#Eighty-eight miRNAs were up-regulated and 46 miRNAs have been reported in colorectal cancer; 40 miRNAs were down-regulated in rectal cancers and 15 miRNAs have been reported in colorectal cancer. To compare the relative miRNA expression levels as measured by RT-qPCR and chip analysis, we analyzed expression levels of these miRNAs in the cancer tissues. The results showed that miRNA expression (increased or decreased) in the paired benign and tumor tissue was consistent between the two methods in all cases. Expression levels of 6 up-regulated miRNAs (by chip analysis compared to RT-qPCR) varied in a range from -11.9% to 39.1% . Expression levels of 5 down-regulated miRNAs varied in a range from 1.4% to 29.4%. The Pearson correlation of relative miRNAs expression levels was analyzed by cDNA array versus RT-qPCR, and found to be 0.96 (P<0.01).@*CONCLUSION@#miRNA profile in rectal cancer showed unique characteristics, and identified a series of new, aberrantly expressed miRNAs.

Hepatocyte-targeted gene transfection of galactosylated chitosan-graft low molecular polyethyleneimine/DNA complexes / 中南大学学报(医学版)

Objective To investigate the hepatocyte targeted specific property of galactosylated chitosan-graft-polyethyleneimine (GC-PEI)/DNA complexes in vitro and in vivo.Methods With the plasmid expressing enhanced green fluorescent protein (pEGFP-C1) as the reporter gene,the formation of GC-PEI/DNA complexes was induced to self-assemble in 0.01 mol/L phosphate buffered saline(PBS),150 mmol/L NaCl,or 5% glucose solution (GS).The complexes were characterized by the particle size,Zeta potential,DNA binding and protection capacity,and further tested for cytotoxicity and hepatocyte targeted degradation of DNaseⅠand the serum,which presented as a well-formed sphere or compacted nucleocapsid structure at a diameter of 50-200 nm.The GC-PEI copolymer showed no obvious toxicity in the tested cell lines.Acute toxicity assay revealed that the mice grew well in 2 weeks with GC-PEI dosage from 50 to 300 μg.The assay by flow cytometry and fluorescent microscope showed that the transfection efficiency in hepatocyte lines (L02,QSG7701/core) was higher than that in non-hepatocyte lines (SGC7901,HBE) in vitro.In vivo,the GFP was obviously expressed in the liver tissue and not expressed in other organs 48 h after the transfection.Conclusion GC-PEI copolymer may carry the exogenous gene specifically to hepatocytes in vitro and in vivo,which has very good liver targeted specific property.

The neuroprotective effects and its mechanisms of qingkailing injection on bacterial meningitis induced by E. coli in rabbits / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To explore the neuro-protective effect and mechanism of qingkailing injection (QKL) against cerebral injury caused by E. coli-meningitis (CM).</p><p><b>METHODS</b>The CM model rabbits were treated by ampicillin with QKL as adjuvant. The leukocyte count and protein content in cerebral spinal fluid (CSF), the contents of water, sodium, potassium and calcium in cerebral tissues were measured before, 16 h and 26 h after Bacillus coli injection respectively. The expression of matrix metalloproteinase-9 (MMP-9) was determined at the same time.</p><p><b>RESULTS</b>Adjunctive treatment with QKL can not only inhibit the increase of leukocyte cells, protein content in CSF, and water, sodium, calcium content in cerebral tissues, but also the decrease of potassium content revealed during simple antibiotic treatment. It also can decrease the expression of MMP-9 in cerebral tissues of rabbits with CM.</p><p><b>CONCLUSION</b>As an adjunctive treatment, QKL can prevent transient inflammatory reaction and aggravation of brain injury in CM induced by simple antibiotic treatment, its mechanisms might relate with calcium antagonism and attenuation of MMP-9 expression in brain tissues.</p>

Hepatitis C virus nonstructural protein NS(3) and telomerase activity / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>To study the effect of hepatitis C virus nonstructural protein NS(3) (HCV NS3) on telomerase activity and carcinogenesis.</p><p><b>METHODS</b>Streptavidin-peroxidase (SP) conjugated method was used to detect the expression of HCV NS(3) protein in NIH3T3 cells transfected with plasmid pRcHCNS(3)-5' and pRcHCNS(3)-3'. Telomerase activity was detected by an in situ telomerase activity labeling method, telomeric repeat amplification protocol polymerase chain reaction (TRAP-PCR) and telomerase PCR enzyme linked immunosorbent assay (ELISA) technology in the transfected and non-transfected NIH3T3 cells.</p><p><b>RESULTS</b>HCV NS(3) protein was expressed in the NIH3T3 cells transfected with plasmid pRcHCNS(3)-5' expressing HCV NS(3) C-terminal deleted protein or with plasmid pRcHCNS(3)-3' expressing HCV NS(3) N-terminal deleted protein. The positive signal of HCV NS(3) protein was localized in the cytoplasm of NIH3T3 cells, and the signal intensity of the former was stronger. Telomerase activity in NIH3T3 cells transfected with plasmid pRcHCNS(3)-5' was stronger than that in NIH3T3 cells transfected with plasmid pRcHCNS(3)-3' (P < 0.01), whereas telomerase activity in NIH3T3 cells transfected with plasmid pRcCMV or untreated NIH3T3 cells was weaker than that in NIH3T3 cells transfected with plasmid pRcHCNS(3)-3' (P < 0.05). The expression level of HCV NS(3) protein was significantly correlated with the strength of telomerase activity (P < 0.05). The results obtained by in situ telomerase activity labeling corresponded to the results by telomerase PCR ELISA technology.</p><p><b>CONCLUSIONS</b>HCV NS(3) protein may activate telomerase through endogenous mechanism to induce host cell transformation. The effect of HCV NS(3) C-terminal deleted protein on telomerase activity in the host cell may be stronger than that of HCV NS(3) N-terminal deleted protein. In situ telomerase activity labeling was a reliable technology for studying pathological morphology and telomerase activity in tissues and cells.</p>

Study of Relationship between Telomerase Activity and Expression of bcl-2 and p53 Proteins in Human Ovarian Epithelial Tumors / 中国医师杂志

Objective To study the relationship between in situ telomerase activity and the expressin of bcl-2 and p53 proteins in human ovarian epithelial tumors(OETs), and explore their effect in pathogenesis of ovarian cystadenocarcinoma. Methods The telomerase activity of ovarian epithelial tumors was measured by in situ telomerase activity labeling (ISLT), and bcl-2 and p53 expression was detected by SP immunohistochemical method. Results ⑴The positive rates of telomerase in ovarian cystadenocarcinomas(OCAC), their surrounding ovarian tissues(SOT),borderline cystadenomas(BCA),and cystadenomas(CA)were 92 3%(24/26), 0(0/26), 42 8%(9/21) and 0(0/15) respectively, and the positive rate was significantly higher in OCAC than that in SOT, BCA and CA(P0 05); ⑵The positive rates of Bcl-2 in OCAC, SOT, BCA and CA were 65 38%(17/26), 0(0/26), 52 38%(11/21) and 26 66%(4/15) respectively, which were significantly higher in OCAC and BCA than those in SOT and CA(P0 05). Conclusion The results indicated that overexpression of bcl-2 protein may be related to telomerase activation, the telomerase activation induced by bcl-2 overexpression may result in malignant transformation of ovarian epithelials, and p53 mutation may not affect telomerase activity during ovarian cystadenocarcinogenesis.

Expression of Vascular Endothelial Growth Factor and nm23 Proteins and Microvessel Density in Hepatocellular Carcinoma and Their Relation With Metastasis / 中国医师杂志

Objective To investigate expressions of vascular endothelial growth factor (VEGF) and nm23 proteins and microvessel density in HCC,simultaneously,their relation with metastasis was also studied.Methods The expression of VEGF and nm23 and CD34 protein in 56 HCCs were detected by immunohistochemical technique.Results Of 56 HCCs, the intensity of VEGF expression in HCCs with metastasis was significantly higher than that in HCCs without metastasis (P