1.Effect of culture supernatant of CpG ODN-stimulated neutrophils from patients with psoriasis vulgaffs on the proliferation of keratinoeytes
Jun SONG ; Meng PAN ; Bangguo LUO ; Guoruan LUO ; Jie ZHENG
Chinese Journal of Dermatology 2009;42(7):481-483
Objective To investigate the role of nentrophils in the pathogenesis of psoriasis vulgaris. Methods Neutrophils were isolated from venous blood samples of 25 patients with psoriasis vulgaris (including 13 cases of active psoriasis and 12 cases of inactive psoriasis) as well as 25 normal human con-trols, and cultured. Then, these neutrophils were grouped and treated with lipopolysaccharide (LPS, 100 g/L),CpG-A (50 mg/L), CpG-B (50 mg/L), and RPMI 1640 culture medium, respectively, for 24 hours followed by the collection of culture supematants. Human keratinocytes (HaCaT) were cultured in the presence of su-pematants of treated or untreated nentrophils for 72 hours followed by the detection of cell proliferation with MTT assay. To determine the role of proinflammatory factors, SOD/CAT and monoclonal antibody to IL-8 and TNF-alpha of 400 u/mL were used to pretreat HaCaT cells 1 hour prior to the stimulation with super-natants of neutrophils. Results Compared with culture medium, the supematant of unstimulated neutrophils from normal controls or patients with inactive psoriasis had no significant effect on the proliferation of HaCaT cells (P > 0.05), but that from patients with active psoriasis markedly promoted the proliferation of HaCaT cells (t = 2.41, P < 0.05). ARe, stimulation by LPS, CpG-A and CpG-B, the supematant of active patient-derived neutrophils significantly promoted the proliferation of HaCaT cells compared with that of normal control-derived nentrophils (t = 3.11, 2.89, 2.29, respectively, all P < 0.05). In comparison with tmstimulated neutrophils, the supematant from LPS- and CpG-A stimulated nentrophiles significantly accelerated the pro-liferation of HaCaT cells. Furthermore, the proliferation of HaCaT cells induced by the supematants of LPS-,CpG-A-, CpG-B-stimulated neutrophils from psoriatic patients was statistically suppressed by the pretreat-ment with the monoclonal antibody to IL-8, TNF-alpha and SOD/CAT (all P < 0.05). Conclusions In patients with psoriasis vulgaris, there is an abnormal secretion of IL-8, TNF-alpha and superoxide by neutrophils in peripheral blood, and these proinflammatory factors could promote the proliferation of HaCaT cells.
2.The role of hydrogen peroxide on mesenteric artery RhoA/ROCK signal pathway in cirrhotic rats with portal hypertension
Ming DUAN ; Dejun LIU ; Jun QIN ; Zhiyong WU ; Meng LUO ; Wei CHEN
Chinese Journal of Hepatobiliary Surgery 2014;20(8):604-608
Objective To explore the role of hydrogen peroxide in mesenteric artery contraction of cirrhotic rats with portal hypertension,which was induced by bile duct ligation.Possible mechanism in RhoA/ROCK signal pathway was also part of the focus.Methods The bile duct ligation-induced cirrhotic rats and normal rats (control group) were treated equally with PEG-catalase(10 000 U/kg-1 · d-1,ip.) or by its vehicle for 8 days.Then the level of H2O2 in mesenteric arteries was detected.The contractile response to norepinephrine of arterioles was analyzed by vascular perfusion system.The protein expressions of the α1 adrenergic receptor,β-arrestin-2 and Rho kinase-1 (ROCK-1),and the activity of ROCK-1 were measured by western blot.In addition,the interaction of α1-adrenergic receptor with β-arrestin-2 was assessed by co-immunoprecipitation.Results Compared to normal rats,the dose-response curve of the mesenteric arterioles in response to norepinephrine shifted to the right,and the EC 50 increased in the rats with portal hypertension.PEG-catalase treatment can decrease the hydrogen peroxide level in arteries,thus significantly lowered EC50 and improved the reactivity to norepinephrine of the mesenteric arterioles in portal hypertension rats.No significant difference in the α1-adrenergic receptor amounts was observed among groups.There was remarkable decreases in the protein expressions of β-arrestin-2 and its interaction with the α1-adrenergic receptor in cirrhotic rats with PEG-catalase treatment.PEG-catalase also increased the amount and activity of ROCK-1 in cirrhotic rats.Conclusions The level of hydrogen peroxide increases in the mesenteric arteries in bile duct ligation-induced cirrhotic rats.And it enhances the β-arrestin-2 expression and its interaction with the α1-adrenergic receptor,which subsequently decreases the amount and activity of ROCK as well as the contractility of mesenteric arteries in response to vasoconstrictors.
3.Combined liver-kidney transplantation in one case
Wenfeng LUO ; Jun SHI ; Dongliang MENG ; Xinchang LI ; Zhidan XU ; Chengmei LONG
Chinese Journal of Tissue Engineering Research 2009;13(5):988-990
A 32-year old male who had liver cirrhosis accompanying with chronic renal insufficiency (uremia) was suffered from homochronous allograft transplantation of liver-kidney. The blood and other tissue typing were in concord between the donor and recipient. The liver transplantation took typical orthotopic transplantation technique while the kidney transplantation took general operation method to place the transplanted kidney at the recipient's right lilac fossa. Daclizumab was taken for the immunity induction treatment before transplantation while the trigeminy of tacrolimus+mycophemocate mofeil+methylprednisolone were taken as immunosuppressant after transplantation. The transplanted liver and kidney recovered well which could work at once without any serious complications after transplantation, suggesting that combined liver-kidney transplantation was an effective treatment method for homochronous function failure of liver and kidney characterizing by perfect tissue typing, excellent operation skill, reasonable usage of immunosuppresant being the key point of success for transplantation.
4.Experimental study of nitric oxide in decreasing intestinal mesenteric arterial hypocontractility in rats with hepatic cirrhosis and portal hypertension
Meng LUO ; Wei CHEN ; Jun QIN ; Dejun LIU ; Jianxia LIU ; Lei QIN ; Haixin QIAN
Chinese Journal of Digestive Surgery 2013;(3):222-227
Objective To investigate the mechanisms of nitric oxide (NO) in decreasing intestinal mesenteric arterial hypocontractility in rats with hepatic cirrhosis and portal hypertension,and to analyze the interaction of NO and RhoA/ROCK pathway.Methods The levels of NO in the peripheral blood and mesenteric artery of normal rats (normal control group,5 rats),rats with portal hypertension (experimental control group,6 rats)and rats with portal hypertension treated by L-NAME (L-NAME group,6 rats) were detected.Mesenteric arteriole contractility to norepinephrine in the 3 groups was determined using a vessel perfusion system.The expressions of proteins of NO-cGMP-PKG pathway and RhoA/ROCK pathway in the 3 groups were detected by Western blot.All data were analyzed using the analysis of variance or LSD-t test.The changes of mesenteric arteriole contractility to norepinephrine was expressed in dose-response curve,and was analyzed using the nonlinear regression method,and the EC50 value was calculated.Results (1) The pressures of portal veins of the normal control group,experimental control group and L-NAME group were (6.2 ± 0.9)mm Hg (1 mm Hg =0.133 kPa),(13.9 ± 1.7)mm Hg and (16.6 ± 1.3) mm Hg,respectively,with a significant difference among the 3 groups (F =94.4,P < 0.05).(2) The levels of NO in the normal control group,experimental control group and L-NAME group were (43 ± 5) μmol/L,(82 ± 16) μmol/L and (45 ± 9) μmol/L,respectively,with a significant difference among the 3 groups (F =24.77,P < 0.05).The level of NO of the L-NAME group was significantly lower than that of the experimental control group (P < 0.05).(3) The levels of NO in the mesenteric artery of the normal control group,experimental control group and L-NAME group were (236 ±41) μmol/g,(407 ± 82) μmol/g and (216 ± 42) μmol/g,respectively,with a significant difference among the 3 groups (F =20.29,P < 0.05).The NO level of the L-NAME group was significantly lower than that of the experimental control group (P < 0.05).(4) Compared with the experimental control group,the dose-response curve of mesenteric arterioles to norepinephrine shifted to the left,while it did not reach the level of the normal control group.The levels of EC50 of the normal control group,experimental control group and the L-NAME group were 6.458 × 10-7 mol,9.546 × 10-7 mol and 7.494 × 10-7 mol,respectively.There was a significant difference in the EC50 level between the L-NAME group and the other two group (t =2.726,3.112,P < 0.05).(5) Compared with the normal control group,the protein expression levels of eNOS and p-VASP of mesenteric artery of the experimental control group were significantly increased (P < 0.05),while they were decreased in the L-NAME group (P < 0.05).The protein expression levels of eNOS and p-VASP of mesenteric artery of the L-NAME group were significantly higher than those of the normal control group (P <0.05).There were no obvious changes of protein expression levels of PKG-1,ROCK-1 and p-moesin in the 3 groups (P > 0.05).(6) The activity of ROCK-1 was significantly increased with norepinephrine stimulation in the normal control group and the L-NAME group,while no obvious changes were detected in the experimental control group.Conclusions The NO expression is upregulated in mesenteric arteries in rats with hepatic cirrhosis and portal hypertension.Such changes induce ROCK activation via influencing the expression of vasoconstrictors.L-NAME can reduce the NO levels in the mesenteric arteries,which may improve RhoA/ROCK signal pathway transduction.This can help vasoconstrictors induce ROCK activation without affecting the protein expression of ROCK.
5.Vascular endothelial growth factor's participation in splanchnic hyperdynamic circulation in portal hypertension
Quanbo ZHOU ; Min ZHANG ; Wei CHEN ; Jun QIN ; Qing XU ; Hong ZHOU ; Meng LUO
Chinese Journal of Hepatobiliary Surgery 2013;19(8):611-616
Objective To detect the variation of vascular endothelial growth factor (VEGF) in the splanchnic vessels under portal hypertension (PHT) and explore its mechanism and influence on the process of hyperdynamic circulation.Methods In humans,the level of VEGF pathway related proteins were detected in the splenic artery of PHT patients in clinical trials.In animal experiments,the following parameters were observed for rats in the control group (group N,n =7) and the CCl4 induced portal hypertension group (group PHT,n=7):portal vein diameter,portal vein blood flow velocity (PBV),portal vein blood flow (PBF),portal vein pressure (PP),norepinephrine (NE) reactivity in the isolated mesenteric artery microcirculation,contractile reactivity and degree of endothelial ni tric oxide synthase (eNOS) activation in the mesenteric artery by selectively inhibiting the VEGF signal pathway with SU5416.In cell experiments,primary culturing of arterial endothelial cells in vitro were used to verify the effects of VEGF on eNOS activation.Results The results showed that VEGF expression levels in the splenic artery of PHT patients significantly increased.In animal experiments,there was not a significant difference in portal vein diameter between group N and group PHT.How ever,the PBV and PBF of group PHT were lower than those of group N,and SU5416 had no clear effect on PBV and PBF in group PHT.PP of group PHT was much higher than that of group N,and SU5416 had little influence on reducing PP in group PHT.The contractile response of mesenteric artery microcirculation to NE in group PHT decreased significantly,EC50 increased a lot,and SU5416 improved this hypoergia partially.The protein levels of VEGF,VEGFR-2,eNOS,and p-eNOS in the mesentery artery of group PHT raised quite a lot compared to group N,and SU5416 decreased the protein level of VEGFR-2 and activation of eNOS significantly.Experiments in vitro confirmed that VEGF could promote the activation of eNOS.Conclusion The excessive VEGF produced in visceral arteries under PHT may participate in the process of hyperdynamic circulation partially through promoting the synthesis and activation of eNOS and then reducing visceral arteries' response to NE.
6.Metric evaluation of quality of life instruments for cancer patients-brain neoplasm (QLICP-BN)
Xiaoyu HAN ; Jun HE ; Chonghua WAN ; Jiahong LUO ; Gang BAI ; Jianghui ZHANG ; Qiong MENG
Journal of International Oncology 2021;48(3):143-149
Objective:To understand the reliability and validity of quality of life instruments for cancer patients-brain neoplasm [QLICP-BN (V1.0)], a self-developed quality of life scale for cancer patients.Methods:The quality of life of 112 patients with brain neoplasms in Yunnan Cancer Hospital from March 2012 to November 2013 was measured. The general data questionnaire and QLICP-BN (V1.0) were used for data collection. The reliability, validity and responsiveness of the scale were tested, and then the metric characteristics of the scale were evaluated.Results:The split-half reliability of the total score of the scale was 0.95, the Cronbach αcoefficient was 0.92, and the test-retest correlation coefficient rwas 0.78. After extracting common factors by the principal component method and rotating with the maximum variance, the specific module obtained three principal components, and the cumulative variance contribution rate was 64.18%. The score of specific module was 75.30±17.44 before treatment and 78.91±12.20 after treatment ( t=-2.481, P=0.015). The total score of scale before treatment was 65.26±12.29, and that after treatment was 69.62±10.41, with a statistically significant difference ( t=-4.492, P<0.001). The total responsiveness of the scale was 0.456, showing moderate responsiveness. Conclusion:QLICP-BN (V1.0) has good reliability, validity and a certain degree of responsiveness. It can be used as a measurement tool for the quality of life of patients with brain neoplasms in China.
7.Safety and short-term effect of antithyroid agents on hyperthyroidism patients coexisting with viral hepatitis.
Tai JIANG ; Kai-Zhong LUO ; Yong-Hong ZHANG ; Hong-Yu LUO ; Jun LIANG ; Meng LIU
Chinese Journal of Hepatology 2010;18(1):59-60
Adolescent
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Adult
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Aged
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Antithyroid Agents
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administration & dosage
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adverse effects
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therapeutic use
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Biomarkers
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blood
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Female
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Hepatitis B
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complications
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pathology
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Hepatitis, Viral, Human
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complications
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pathology
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Humans
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Hyperthyroidism
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complications
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drug therapy
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Liver Function Tests
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Male
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Methimazole
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administration & dosage
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adverse effects
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therapeutic use
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Middle Aged
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Propylthiouracil
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administration & dosage
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adverse effects
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therapeutic use
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Retrospective Studies
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Severity of Illness Index
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Thyroid Function Tests
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Young Adult
8.Gene-gene interactions among the peroxisome proliferator-activated receptor polymorphisms for hypertriglyceridemia.
Shu-jun GU ; Meng-meng LIU ; Zhi-rong GUO ; Ming WU ; Qiu CHEN ; Zheng-yuan ZHOU ; Hao YU ; Li-jun ZHANG ; Wen-shu LUO
Chinese Journal of Preventive Medicine 2012;46(10):916-921
OBJECTIVETo investigate the association of ten SNP at peroxisome proliferator-activated receptors (PPARα, δ, γ) with hypertriglyceridemia and the gene-gene interaction.
METHODSParticipants were recruited from the Prevention of MetS and Multi-metabolic Disorders in Jiangsu province of China Study (PMMJS). A total of 820 subjects were selected from the 4083 participants who had received follow-up examination, by using simple random sampling. Participants in baseline and follow-up study surveys were both collected blood samples 11 ml in the morning after at least 8 hours of fasting. Blood samples which collected at the baseline were subjected to PPARα, PPARδ and PPARγ genotype analyses. Blood samples which collected at the follow-up were used to measure serum triglyceride levels. The logistic regression model was used to analyze the association between different SNP and hypertriglyceridemia, and the generalized multifactor dimensionality reduction (GMDR) was applied to explore the gene-gene interaction.
RESULTSThe samples included 474 in the non-hypertriglyceridemia group and 346 in the hypertriglyceridemia group. The genotype frequencies of rs1800206 in the hypertriglyceridemia group were 211 (61.0%) for LL, 132 (38.2%) for LV and 3 (0.9%) for VV, and in the non-hypertriglyceridemia group were 411 (86.7%) for LL, 59 (12.4%) for LV and 4(0.8%) for VV (χ(2) = 74.18, P < 0.01). V allele frequencies of rs1800206 in the hypertriglyceridemia group was 138(19.9%), and in the non-hypertriglyceridemia group was 67 (7.1%) (χ(2) = 60.62, P < 0.01). The genotype frequencies of rs2016520 in the hypertriglyceridemia group were 177 (51.2%) for TT, 154 (44.5%) for TC and 15 (4.3%) for CC, and in the non-hypertriglyceridemia group were 211 (44.5%) for TT, 212 (44.7%) for TC and 51 (10.8%) for CC(χ(2) = 15.93, P < 0.01). C allele frequencies of rs2016520 in the hypertriglyceridemia group was 184(26.6%), and in the non-hypertriglyceridemia group was 314 (33.1%) (χ(2) = 8.07, P < 0.01). The genotype frequencies of rs3856806 in the hypertriglyceridemia group were 149 (43.1%) for CC, 156 (45.1%) for CT and 41 (11.8%) for TT, and in the non-hypertriglyceridemia group were 269 (56.8%) for CC, 170 (35.9%) for CT and 35 (7.4%) for TT (χ(2) = 15.93, P < 0.01). T allele frequencies of rs3856806 in the hypertriglyceridemia group was 238(34.4%), and was 240 (25.3%) in the non-hypertriglyceridemia group (χ(2) = 15.96, P < 0.01). The genotype frequencies of rs1805192 in the hypertriglyceridemia group were 145 (41.9%) for PP, 158(45.7%) for PA and 43(12.4%) for AA, and in the non-hypertriglyceridemia group were 314 (66.2%) for PP, 137(28.9%) for PA and 23(4.9%) for AA (χ(2) = 50.92, P < 0.01). A allele frequencies of rs1805192 in the hypertriglyceridemia group was 244(35.2%), and was 183 (19.3%) in the non-hypertriglyceridemia group(χ(2) = 52.89, P < 0.01). After adjusting age, gender, smoking, alcohol consumption, high-fat diet, low -fiber diet and occupational physical activity factors, rs1800206, rs2016520, rs3856806 and rs1805192 were significantly associated with hypertriglyceride, while the OR (95%CI) was 3.88 (2.69 - 5.60), 0.71 (0.52 - 0.96), 1.40 (1.03 - 1.90) and 2.56 (1.88 - 3.49), respectively (P < 0.05). GMDR model analysis showed that the second-order model (rs1800206 and rs1805192) was the best model when quality traits of triglyceride was chosen as outcome (P < 0.01); while third-order model (rs1800206, rs1805192 and rs2016520) was the best model when quantitative traits of triglyceride was chosen as outcome (P < 0.01).
CONCLUSIONThe rs1800206, rs2016520, rs3856806 and rs1805192 were significantly associated with hypertriglyceridemia. There was a gene-gene interaction between multiple SNP.
Adult ; China ; Female ; Gene Frequency ; Genotype ; Humans ; Hypertriglyceridemia ; blood ; genetics ; Logistic Models ; Male ; Middle Aged ; Peroxisome Proliferator-Activated Receptors ; genetics ; Polymorphism, Single Nucleotide
9.Colorimetric detection of norovirus genotype GII by reverse transcription loop-mediated isothermal amplification.
Jian-Ming LUO ; Xi-Yang WU ; Zi-Qian XU ; Le LUO ; Kai NIE ; Meng-Jie YANG ; Ya-Lan ZENG ; Zhao-Jun DUAN ; Xue-Jun MA
Chinese Journal of Virology 2012;28(2):165-171
A simple, rapid and sensitive colorimetric reverse transcription loop-mediated isothermal amplification (RT-LAMP) method was established to detect norovirus genotype GII. The method employed a set of six specially designed primers that recognized eight distinct sequences of RNA-dependant RNA polymerase and capsid protein gene for amplification of nucleic acid under isothermal conditions at 65 degrees C for 60 minutes. The amplification process of RT-LAMP was monitored by the addition of HNB (Hydroxy naphthol blue) dye prior to amplification. A positive reaction was indicated by a color change from violet to sky blue and confirmed by agarose electrophoresis. The specificity of the RT-LAMP was validated by detecting several different diarrhea viruses including norovirus genotype GII. The sensitivity was determined by serial dilutions of RNA molecules from in vitro transcription of norovirus genotype GII in parallel with conventional RT-PCR detection. The assay was further evaluated with 93 clinical specimens of diarrhea patients. The results showed that the sensitivity of RT-LAMP was 1 000 copies/microL with a high specificity and the relative sensitivity was at the same level as that of conventional RT-PCR. Positive rate of RT-LAMP in analysis of clinical specimens was approximately the same as that of conventional RT-PCR as well. This colorimetric RT-LAMP assay was potential for rapid detection of norovirus genotype GII on spot due to the observation of visual result with high specificity and sensitivity, time-saving and cost benefit.
Caliciviridae Infections
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diagnosis
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virology
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Colorimetry
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methods
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Feces
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virology
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Genotype
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Humans
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Norovirus
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genetics
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isolation & purification
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Nucleic Acid Amplification Techniques
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methods
10.Effect of nitric oxide on HaCaT cell migration.
Shi-wei YANG ; Jun WU ; Gao-xing LUO ; Xiao-rong ZHANG ; Xiao-hong HU ; Yan-meng PENG ; Jun-jie YANG ; Xiao-li LUO ; Ying WANG
Chinese Journal of Burns 2010;26(2):146-149
OBJECTIVETo investigate the effect of exogenous nitric oxide (NO) on the migration of HaCaT cell and its possible mechanism.
METHODSSodium nitroprusside (SNP) was used as the donor of NO. Different concentrations of SNP (0.1, 1.0, 10.0, 100.0, 1000.0 micromol/L) were added into nutrient culture medium of HaCaT cells. Cell migration rate was observed and calculated at post scratching hour (PSH) 0 (immediately after scratching), 6, 12, 24, 48. The most suitable concentration of SNP and culture duration were selected as stimulation condition. Cytoskeletons of HaCaT cells were observed under confocal laser scanning microscope. The expressions of integrin beta 1, RhoA, Rac1 and Cdc42 of cells in experiment group (cultured with 10.0 micromol/L SNP for 24 hours) and negative control group were determined at mRNA and protein levels with RT-PCR and Western blot respectively. Data were processed with one-way analysis of variance (ANOVA) and repeated measure ANOVA.
RESULTSMigration rate of HaCaT cells in each group increased gradually as time after scratching went on. There were significant differences between PSH 6-48 and PSH 0 in cells cultured with 10.0 micromol/L SNP (F = 31.002, P values all below 0.05). Pili were rarely observed in negative control group with slender stress fibers in cells. In comparison, the amount of pili amount increased obviously in experiment group with thickened stress fibers. Compared with those of cells in control group (RhoA protein expression = 0.64 +/- 0.04), integrin beta 1 expression decreased obviously (F = 8.25, P = 0.015), RhoA (0.92 +/- 0.04), Cdc42 and Rac1 were up-regulated at both protein (with F value respectively 7.25, 14.10, 6.50, P values all below 0.05) and mRNA levels (with F value respectively 23.67, 10.39, 9.52, P values all below 0.05).
CONCLUSIONSExogenous NO in suitable concentration can promote the proliferation and migration of HaCaT cell, suggesting it exerts significant effect in wound repair. The changed cytoskeletons and the down-regulated integrin beta 1 expression may be involved in this process.
Cell Line ; Cell Movement ; drug effects ; Cytoskeleton ; drug effects ; metabolism ; Humans ; Nitric Oxide ; pharmacology ; RNA, Messenger ; genetics ; rhoA GTP-Binding Protein ; genetics ; metabolism