Manual reduction combined with percutaneous kyphoplasty for fresh osteoporotic vertebral compression fractures in elderly patients / 医学研究生学报

Objective Minimally invasive treatment of orthopedic diseases is the general direction of future development of medicine.This study was designed to observe the effect of manual reduction combined with percutaneous kyphoplasty (MR+PKP) in the treatment of fresh osteoporotic vertebral compression fractures ( OVCF) in elderly patients. Methods Sixty OVCF patients aged 60-86 ( mean 72.3) years were randomly assigned to 2 groups of e-qual number to be treated by MR+PKP and PKP alone, respectively. Comparisons were made between the two groups of patients in the op-eration time, volumeand permeability of the bone cement injected,changes of the Cobb angle,restoration of the anterior height of the compressed vertebral bodies,pre-and post-operative Visual Analogue Scale ( VAS) pain scores, OswestryDisability Indexes ( ODIs) , and other differences observed before and aftersurgery. Results Op-erations were performed successfully in all the 60 cases.In the MR+PKP group, the mean operation time was 61 min, the mean volume of bone cement injected was 5.1mL with qualified distribution, and bone cement leakage occurred in 1 case without adverse reaction. Statistically significant differences were found in the pre-and post-operativeanterior height of the compressed vertebral bodies, Cobb an-gle, VAS scores, and ODIs (P<0.05).Compared with the PKP control, MR+PKP achieved a significant increase at 3 days and 3 months after surgery in the anterior height of the compressed vertebral bodies ([22.4±1.4] vs [26.8±8.1] mm and [21.4±4.2] vs [26.5±7.2]mm, P<0.05), and a decrease in the Cobb angle ([8.6±2.7] vs [8.1±2.1]°and [9.0±2.3] vs [8.3±1.8]°, P<0.05) as well as remarkably reduced VAS scores (4.1±2.2vs 3.1±2.0, P<0.05)and ODIs (23.0±3.1vs25.6±3.3, P<0.05) at 3 d postopera-tively. Conclusion MR+PKP, with its advantages of effective pain-relief, improvement of the height of compressed vertebral bodies, and reduction of bone cement leakage,is better than PKP alone for the treatment of OVCF in elderly patients.

Isolation and purification of antimicrobial polypeptide HMGN2 from human lymph node and analysis of its distribution / 生物医学工程学杂志

This study was conducted to isolate and purify antimicrobial polypeptides HMGN2 (high mobility group nucleosomal-binding domain2) from human lymph node, to detect the antimicrobial activity of HMGN2, and to determine the subcellular location of HMGN2 in human lymph node. The antimicrobial polypeptides were purified by the Reverse Phase HPLC and identified by Tricine-SDS-PAGE. The antimicrobial activity was detected by agar diffusion test. Mass spectrum and Western-blot analysis indicated the individual character of protein. HMGN2 was isolated and purified from human lymph node, and it showed antimicrobial potency against the pathogenic strain E. coli 54,080. The immunocytochemistry staining indicated that HMGN2 was present both in human lymph node cells' nucleus and cytoplasm. In conclusion, HMGN2 protein is of antimicrobial activity and it is probably involved in the defence of innate immunity in vivo.

Transfection of dominant negative MyD88 decreases IL-8 production in bacteria-infected airway epithelial cells / 生物医学工程学杂志

Interleukin-8 (IL-8) is an important activator and chemoattratant of neutrophils and has been implicated in airway inflammatory diseases. To explore the new gene therapeutic strategies for airway inflammation, plasmid expressing dominant negative myeloid differentiation protein (MyD88 DN) was constructed and transfected into human airway epithelial cell lines A549 and SPC-A-I. The cells were challenged with M. tuberculosis, P. aeruginosa or K. pneumoniae and the release of IL-8 was measured using ELISA. The results showed that the supernatants of M. tuberculosis and R. aeruginosa enhanced IL-8 release from the epithelial cells; and transfection of MyD88 DN diminished this effect. MyD88 DN also reduced IL-8 release from cells induced by live bacteria of P. aeruginosa or K. pneumoniae. These data suggest that MyD88 could be used as a target gene in the gene therapy of airway inflammation.

Construction and identification of HBD-2 transgenic mice / 生物医学工程学杂志

Human beta defensin 2 (HBD-2) may play an important role in human defense against infection. Its antimicrobial capacity has been fully documented in in vitro study. In order to evaluae its in vivo effects, we developed an HBD-2 transgenic mouse model. The HBD-2 minigene containing CMV promoter, full length of HBD-2 cDNA and BGH polyA tail was generated by PCR amplification and introduced into the fertilized oocytes of C57 X ICR hybridized mouse by microinjection, and offspring were produced. DNA was isolated from the tails of the mouse pups, and the HBD-2 minigene incorporation was analyzed by PCR using HBD-2 specific primers. The HBD-2 gene expression in the multi-tissues of transgenic mice was determined at mRNA level by RT-PCR and at peptide level by immunohistological staining with the use of HBD-2 monoclonal antibody. The results showed that among 17 F0 transgenic mice, HBD-2 positive signal was determined by PCR in 4 mice, suggesting that HBD-2 minigene has been incorporated into the offspring mice. Meanwhile, a widespread expression of HBD-2 mRNA and peptide was detected in the F1 transgenic mice's multi-tissues such as trachea, lung, intestine, esophagus, testis, spleen, skin, endothelium and brain.

Construction of HNP-3 Mature Peptide Bait Plasmid of Yeast Two-hybrid System and Detecting Its Self-activating and Toxic Effect / 航天医学与医学工程

Objective To construct the bait plasmid of HNP-3 mature peptide in yeast two-hybrid system and examine whether the recombinant bait plasmid has self-activating and toxicity effect.Methods Using RT-PCR technique,the cDNA fragments of HNP-3 mature peptide gene were amplified from the extracted RNA in cultured HL-60 cells.The fragment was firstly cloned into pBluescript-SK-II vector,confirmed by sequencing,then sub-cloned into the bait plasmid pGBKT7 and identified with PCR and sequence analysis techniques.The recombinant plasmid was introduced into the yeast cell AH109,and its self-activating and toxicity effect was tested by auxotrophic selective culture.Results DNA sequencing indicated that the inserted fragment in pBluescript-SK-II vector was HNP-3 mature peptide gene sequence,and the sub-cloned recombinant pGBKT7-HNP-3 was no mismatch.The recombinant bait plasmid didn't have self-activating effect and did not show toxicity to yeast AH109 cell.Conclusion The bait plasmid of HNP-3 mature peptide was constructed successfully.This was helpful for investigating the proteins interacting with HNP-3 mature peptide by yeast two-hybrid technique.

Production of polyclonal antibody against HBD-2 by immunization with recombinant GST-HBD-2 fusion protein / 生物医学工程学杂志

For the purpose of detecting the HBD-2 expression at protein level, the recombinant prokaryotic expression vector pGEX-1lambdaT-HBD-2 was constructed and the E. coli-based product of GST-HBD-2 fusion protein was prepared. When rabbit was immunized with the fusion protein, the anti-serum against HBD-2 was produced. After caprylic acid and ammonium sulfate precipitation, high titer of specific polyclonal antibody against HBD-2, which was detected by ELISA and Western blot, was obtained. This result suggests that recombinant peptide fusion protein could be used instead of the conjugate of peptide-albumin or peptide-thyroid globulin to produce antibody. The obtained antibodies could be used for revealing the tissue distribution of HBD-2 and the regulation of its gene expression.

Construction and immunological study of recombinant hBD-2/PSMA chimeric protein eukaryotic expressive plasmid / 生物医学工程学杂志

The recombinant PSMA DNA vaccine for active immunotherapy of prostate cancer was investigated. Two DNA vaccine recombinant plasmids, pcDNA3.1/PSMA and pcDNA3.1/hBD-2-PSMA, were constructed by inserting the hBD-2 gene and PSMA gene into an eukarytoic expression vector pcDNA3.1. Expression of the two recombinants was detected in transfected COS-7 cells and inoculated mouse muscular cells by RT-PCR and immunohistochemical method. When immunized with pcDNA3.1/PSMA and pcDNA3.1/hBD-2-PSMA, the immunized BALB/c mice acquired specific antibody and T cell response to PSMA. The quantity of the spleen lymphocytes and their CTL activity against PSMA gene transfected-BALB/3T3 cells significantly increased in the immunized mice, and the CTL activity of lymphocytes from pcDNA3.1/hBD-2-PSMA immunized mice was significantly higher than that of pcDNA3.1/PSMA immunized mice. This result suggests that pcDNA3.1/hBD-2-PSMA would probably be developed as a DNA vaccine for the immunotherapy of prostate cancer.

E. coli-based production of recombinant HMG-17 and its antibacterial domain / 生物医学工程学杂志

Total RNA was extracted from human LAK cell, and a cDNA encoding mature peptide HMG-17 and its alpha helix domain was amplified by RT-PCR. The recombinant prokaryotic expression vector pGEX-1lambdaT-HMG-17 and pGEX-1lambdaT HMG-17alpha helix was constructed. Using affinity chromatography, thrombin cleaving and AU-PAGE elution, we obtained the purified HMG-17. Analyses of MIC, MEC and MBC indicated that HMG-17 and HMG-17alpha had strong antibacterial activity. MIC of the alpha-helic domain was almost the same as that of HMG17, suggesting that the alpha-helic structure would be essential for the antibacterial activity of HMG-17.

Application of HMGN2-tag constructs to analysis of HMGN2 distribution in HeLa cells / 生物医学工程学杂志

This study sought to clarify the distribution of HMGN2 in HeLa cells. The recombinant eukaryotic expression vectors pcDNA3. 1-myc-his-HMGN2 and pEGFP-N1-HMGN2 were constructed, and then were transfected into HeLa cells. immunocytochemistry staining indicated that HMGN2 were present not only in HeLa nucleus but also in the cytoplasm. The presence of HMGN2 was also detected in the culture supernatant by ELISA with rabbit anti-serum against HMGN2 and mouse anti-His6 monoclonal antibodies. The confocal microscope observation showed the same subcellular localization as that of immunocytochemistry staining. There results suggested that HMGN2 could be present in the nucleus and cytoplasm of HeLa cell as well as in the extracellular environment.

Airway beta-defensin-2 gene transfer enhances the bacterial clearing of rat lung / 生物医学工程学杂志

beta-defensins possess a broad spectrum of antimicrobial activity. In this study its in vivo antibacterial effect was evaluated by using gene transfer. Rat beta-defensin-2 (rBD2) recombinant pBK-CMV-rBD2 and pCD-NA-3, 1-Myc-His(+)-rBD2 were constructed. Then, by use of liposome agent, the recombinants were delivered into rat airway via tracheal injection. The rBD-2 mRNA expression was detected in the trachea by RT-PCR and its protein expression was determined in the lungs by the tag His immunostaining, 24 hours after inoculation via trachea, the count of P. areuginosa in the lung of rat transfected with pBK-CMV-rBD2 markedly decreased, compared with the control (n = 8, P = 0.003). The data presented in this study provide evidence that airway beta-defensin-2-gene transfer can protect the rat against bacterial infection in vivo, suggesting the beta-defensins as part of the innate host defense system can be of potential applicability.

E. coli-based production of recombinant FALL-39 / 生物医学工程学杂志

This study was aimed at constructing a prokaryotic expression system to resolve the difficulties in acquiring antibacterial peptide and to meet the needs of research and drug development. Total RNA was extracted from human pulmonary gland epithelial cell line SPC-A-1, and a cDNA encoding mature FALL-39 peptide was amplified by RT-PCR. The recombinant prokaryotic expression vector pGEX-1 lambda T-FALL-39 was constructed. Using affinity chromatography, thrombin cleaving and AU-PAGE elution, we obtained the purified FALL-39. MIC, MEC, MBC analyses demonstrated that the FALL-39 had strong antibacterial activity.

Expression of recombinant human beta-defensin-2 gene with C terminal of double marks of Myc and poly-histidine in transfected COS-7 cells / 生物医学工程学杂志

The aim of this study is to explore the possibility and technical itinerary of establishing an mammal engineering cell line in which hBD-2 can be effectively expressed, secreted, detected, separated and purified. The full hBD2 cDNA was inserted into the multi clone site of a eukaryotic expressive plasmid pcDNA3.1/Myc-His(+) and located closely at the upstream of two tag gene (myc and 6 Poly-histidines) so as to construct another recombinant eukaryotic expressive vector of hBD-2 gene: rpcDNA3.1/Myc-His/hBD-2. By the use of RT-PCR with special primers, a band of 240 bp was amplified from COS-7 cells transfected by this recombinant plasmid, which matched full length of cDNA coding hBD2 plus myc epitope and 6 poly-histidines tags. Western blot analysis with specific anti-histidines antibody revealed that the lysate of COS-7 cells transfected by rpcDNA3.1/Myc-His/hBD-2 had a strong band with molecular weight of about 10 Kd that was approximate to the size of chiasmic peptide. Antibacterial activity assay showed that obvious bacterial inhibition occurred in both lysate and supernatant of COS-7 cells transfected by rpcDNA3.1/Myc-His/hBD-2.

Assessment of the role of TLR-4 in shear-stress-induced IL-8 gene transcription activation in vascular endothelial cells by gene mutation and gene transfection technology / 生物医学工程学杂志

This study is to evaluate the role of TLR-4 in lower laminar shear stress-induced interleukin-8 gene transcription activation in vascular endothelial cells by using gene mutation and transfection techniques. RT-PCR, Northern hybridization and immunocytochemical fluorescent staining showed that TLR-4 was expressed in human umbilical vein endothelial cells(HUVEC). When stimulated with 4.2 dyne/cm2 shear stress for 1 hour, an increase of TLR-4 mRNA expression was observed in HUVECs detected by RT-PCR and Northern hybridization. The intracellular domain deletion mutant TLR-4 cDNA (lacking the 155 COOH terminal amino acids of the wild type (TLR-4) and -102 -61 bp DNA sequence in 5'-flanking region of IL-8 gene (IL-8USCS) were isolated from endothelial cells by RT-PCR and PCR. These two DNA fragments were cloned into pcDNA3 and pEGFP1 respectively to construct TLR-4 dominant negative mutant pcDNA3-mTLR4 and IL-8 reporter gene pEGFP1-IL8USCS. ECV304 cells were transfected with the pEGFP1-IL8USCS or co-transfected with pEGFP1-IL8USCS and pcDNA3-mTLR4 by Dosper liposome transfectional reagent and selected by G418, then stimulated with 4.2 dyne/cm2 shear stress for 3 hours. Flow cytometric analysis showed that when exposed to 4.2 dyne/cm2 shear stress for 3 hours, there was a marked increase in the green fluorescent protein expression in pEGFP1-IL8USCS-transfected ECV304 cells. In contrast, there was almost no change in the green fluorescent protein expression in the cells co-transfected with pEGFP1-IL8USCS and pcDNA3-mTLR4 after the stimulation, suggesting the TLR-4 mutant depressed TLR-4 signaling. These experiments suggest that the inflammatory TLR-4/NF-kappa B signaling pathway would probably be involved in flow shear stress-induced IL-8 gene expression in human vascular endothelial cells.

Shear stress induces interleukin-8 mRNA expression and transcriptional activation in human vascular endothelial cells / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>To examine interleukin-8 (IL-8) mRNA expression induced by flow shear stress in human umbilical vein endothelial cells (HUVECs) and investigate its transcriptional activation.</p><p><b>METHODS</b>Flow laminar shear stress 4.2 dyne/cm(2) was used for the stimulating experiments. The flow shear stress-induced IL-8 mRNA expression was detected by reverse transcription-polymerase chain reaction (RT-PCR). pEGFP1 was used to construct IL-8 reporter gene pEGFP1-IL8USCS for determining IL-8 gene transcriptional activation through gene transfer and flow cytometric analysis. NF-kappaB nuclear translocation was observed by immunocytofluorescent staining. Western blot was used to examine IkappaB phosphorylation and degradation. RT-PCR, Northern blot, immunocytofluorescent staining and laser confocal microscopy were used to determine Toll-like receptor-4 (TLR-4) expression at mRNA and protein levels in the cells.</p><p><b>RESULTS</b>There was a marked increase in IL-8 mRNA expression in HUVECs after 120 min of exposure to laminar flow shear stress. When exposed to shear stress for 180 min, there was an increase in enhanced green fluorescent protein expression in pEGFP1-IL8USCS-transfected endothelial cells. NF-kappaB p65 immunocytofluorescent staining of HUVECs showed that when exposed to the same flow shear stress for 30 or 60 min, the cell nuclei became stained; after 90 or 120 min, the staining became much more pronounced. A significant increase in P-IkappaB in the cell lysates occurred after 10 min of exposure while blot density dramatically dropped after 60 min of exposure. The density of the IkappaB blot dropped with increasing exposure time after 30 min. TLR-4 was present on the surface of HUVECs. HUVECs constitutively expressed TLR-2 and TLR-4 mRNA; when exposed to flow shear stress for 60 min, TLR-4 mRNA expression increased.</p><p><b>CONCLUSION</b>NF-kappaB activation is involved in flow shear stress-induced IL-8 mRNA expression in human umbilical vein endothelial cells. TLR-4 receptor for innate immunity most likely mediate these events.</p>

Study of β-defensin rBD-2 gene expression in the pulmonary tissues of the fetal, neonatal and adult rats / 中国病理生理杂志

AIM: To investigate the developmental regulation of β-defensin rBD-2 gene expression in the rat lung. METHOD: Total RNA was isolated from the pulmonary tissues of the fetal, neonatal and adult rats. RT-PCR were performed with primers (P1: TTCAGTCATGAGGATCCATT AC; P2: TGGAACTTGGTCTTTTTATCTAC). The RT-PCR products were cloned into pGEM-T easy vector and the recombinant plasmid was analyzed with EcoR1 digestion and the inserted DNA sequencing was performed on ABI PRISM-377 DNA sequencer. RESULTS： Rat β-defensin-2 transcripts were detected in all the pulmonary tissues of rats during different developmental stages, e.g. at just before birth, 8 hours and 4 days after birth , and adult. CONCLUSION： The rat β-defensin-2 is constitutively expressed in the pulmonary tissues, suggesting that β-defensin-2 may play a role in the lung innate defense against infection.

Toll receptor signaling pathway in inflammation / 中国病理生理杂志

Toll signaling pathway may play a curcial role in induction of inflammation-associated gene activation. Originally, the Toll/spaetzle/Cactus-Dorsal signaling pathway is established in the Drosophila embryonic development. Recently, the Toll signaling pathway in adult Drosophila has been established in the induction of antimicrobial peptide expression. Five human Toll-like receptor genes (h Tlr l-5 ) and one mouse Toll-like receptor gene (m Tlr-4 ) have been isolated. Toll and Toll-like receptor genes encoded molecules are transmembrane proteins with an extracellular leucine repeat domain and a cytoplasmic domain homologous to IL-1 receptors. The intracellular signaling cascade involves Tube, Pelle, and Cactus-Dorsal complex in Drosophila, and MyD88, IRAK, TRAF 6, NIK, ??-I ?B kinase, and I ?B -NF?B complex in mammals. Dorsal and NF?B are transcription factors, while Cactus and I?B are their inhibitors. When the inhibitors phosphorylated, the nuclear factors are released and move into nucleus, leading to immune gene activation. It has been shown from in vitro system that Tlr -4 mediated LPS signaling in human monocytes for expression of IL-1, IL-6, IL-8, and costimulator B7-1 which provides second signal for T cell response. Tlr -2 can also mediate LPS signaling in human monocytes, leading to the production of proinflammatory mediators. Microbial lipoproteins are potent stimulators of IL-12 production through Tlr -2 signaling by human macrophages, and can stimulate Tlr 2-dependent transcription of inducible nitric oxide synthase and the production of nitric oxide, a powerful microbicidal pathway. Findings of a point mutation of Tlr-4 in LPS tolerant C3H/HeJ mouse strain and a deletion of Tlr-4 in LPS resistant C57BL/10ScCr mice provide an in vivo evidence strongly supporting the crucial role of Tlrs in LPS mediated inflammation. It is proposed that targeting Tlrs would develop new remedies for treatment of inflammatory disorders and for immunotherapy of mucosal infections and cancer, etc.

Cytotoxicity to HL-60 cell line and direct viral inactivation of neutrophil defensins / 中国病理生理杂志

A number of low molecular weight cationic peptides, named defensins,were purified from neutrophil granuals. Its in vitro cytotoxicity to HL-60 cell line anddirect viral inactivation were examined. The results showed that when incubated with NP-1 and NP-2(100?g/ml) fot 6 hours, more than 60% of HL-60 cells were killed. WhenHerpes simplex virus type I(HSy-I) and influenza virus B strain Beijing 87-1 wereincubated with NP-1(260?g/ml) at 37?C in water bath for 60 min, a marked reduction(98.2% and 90% ) in virus TCID_50 was observed. The results suggested that neutrophilgranulocyte might be able to synthesize peptides which have the cytotoxic and antiviralacitivities.

Study of ?-defensin rBD-2 gene expression in the pulmonary tissues of the fetal, neonatal and adult rats / 中国病理生理杂志

AIM: To investigate the developmental regulation of ?-defensin rBD-2 gene expression in the rat lung. METHOD: Total RNA was isolated from the pulmonary tissues of the fetal, neonatal and adult rats. RT-PCR were performed with primers (P 1: TTCAGTCATGAGGATCCATT AC; P 2: TGGAACTTGGTCTTTTTATCTAC). The RT-PCR products were cloned into pGEM-T easy vector and the recombinant plasmid was analyzed with EcoR1 digestion and the inserted DNA sequencing was performed on ABI PRISM-377 DNA sequencer. RESULTS: Rat ?-defensin-2 transcripts were detected in all the pulmonary tissues of rats during different developmental stages, e.g. at just before birth, 8 hours and 4 days after birth , and adult. CONCLUSION: The rat ?-defensin-2 is constitutively expressed in the pulmonary tissues, suggesting that ?-defensin-2 may play a role in the lung innate defense against infection.

Rat ?-defensin rBD-1 gene expresses constitutively in skin and kidney / 中国病理生理杂志

AIM: To determine tissue distribution of rat ?-defensin rBD-1 gene expression.METHODS: Total RNA was isolated from 10 kinds of rat tissues. RT-PCR were performed with primers (R 1 5′→3′ ACTCTGGACCCTGACTTCACCG; R 2 5′→3′ CCCTTGCTTGTCCTTTATGTCC). The RT-PCR products around 272 bp in size were cloned into pGEM-T easy vector and the recombinant clones were analyzed by digestion with restriction endonucleases and DNA sequencing.RESULTS: Rat ?-defensin rBD-1 transcripts were found in the kidney and skin, whereas its mRNA was not detected in trachea, uterus, bladder, small intestine, spleen, skeletal muscle, bone marrow and parotid. Sequence analysis confirmed that the RT-PCR product is rBD-1 cDNA. CONCLUSION: These data suggested that ?-defensin rBD-1 may participate not only in the kidney but also in the skin natural defense against infections.

Involvement of TLR-4 signaling pathway in shear stress induced IL-8 gene expression in ECV304 cells / 中国病理生理杂志

AIM: To examine the role of TLR-4 signaling pathway in laminar low shear stress-induced IL-8 gene expression in ECV304 cells. METHODS: RT-PCR and PCR were used to amplify a TLR4 mutant (lacking the 155 COOH terminal amino acids of the wild type TLR4 ) and-102-+61 bp 5′-flanking region of IL-8 gene (IL8 USCS)from endothelial cells. These two DNA fragments were cloned into pcDNA3 and pEGFP1, respectively, and the recombinant plasmid pcDNA3-mTLR4 and pEGFP1-IL8USCS were obtained. ECV304 cells were transfected with the pEGFP1-IL8USCS or co-transfected with pEGFP1-IL8USCS and pcDNA3-mTLR4 by using Dosper liposomal transfectional reagent and selected by G418, and then stimulated by 4 2 dyne/cm 2 shear stress for 3 hours. The green fluorescent protein expression was analyzed by Flow Cytometry. Immunoblotting of the cell lysates and NF-?B p65 immunocytofluorescent staining were used to determine I?B phosphorylation, degradation and NF-?B activation. RESULTS: Flow Cytometric analysis showed that when exposed to 4 2 dyne/cm 2 shear stress for 3 hours, there was a marked increase in the enhanced green fluorescent protein expression in pEGFP1-IL8USCS-transfected ECV304 cells. In contrast, there was almost no change in the green fluorescent protein expression in the cells co-transfected with pEGFP1-IL8USCS and pcDNA3-mTLR4 after the stimulation. Western blot analysis showed that a markedly increased p-I?B of cell lysates occurred at 10 min of exposure and the blot density almost dropped down to the baseline after 60 min of exposure. The density of I?B blot dropped down with increasing exposure time. NF-?B p65 immunocytofluorescent staining of ECV304 cells showed that when exposed to the same flow shear stress for 0 5,1 hours, the cell nuclei became staining, and after 1 5 or 2 hours, the staining was very strong. CONCLUSION: These results suggested that the inflammatory TLR-4/NF-?B signaling pathway may be involved in flow shear stress-induced IL-8 gene expression in human vascular endothelial cells.