2.Promotion of resveratrol on psoriasis like skin damage by down-regulating expression of Keratin 17
Ming WEI ; Lu GAN ; Jin HOU ; Lihong CHEN ; Yantong LIU
Drug Evaluation Research 2017;40(1):37-41
Objective To observe the therapeutic effect of resveratrol (Rsv) on psoriasis like skin damage induced by imiquimod (IMQ) and study its mechanism.Methods BALB/c mice were randomly divided into four groups:control group,Rsv group,model (IMQ) group and Rsv treatment (IMQ + Rsv) group with six mice in each group.Psoriasis like skin model induced by smearing IMQ in the mice's ear for 8 d,20 mg Rsv cream were gave at the same time for treatment.Observe erythema and psoriasis degree,and measure ear thickness with vemier caliper.HE staining was used to observe tissue change,real time quantitative PCR (qRT-PCR) was used to detect K17,IL-17A,IL-19 and IL-23 mRNA and Western blotting were used to detected K17 protein expression in tissues.Results Compared with the model group,mice erythema area and the thickness of epidermal scale decreased significantly in Rsv treatment group;HE staining results showed that,Rsv significantly reduced ear marginal tissue epidermal thickness,and diminished severity of the psoriasis-like skin inflammation.qRT-PCR and Western blotting confirmed a Rsv dependent decrease of in K17,IL-17A and IL-19 in mRNA level and protein levels of K17.Conclusion Rsv improves psoriasis like skin damage by down-regulating expression of Keratinl 7.
3.Associations of serum uric acid, bilirubin levels and short-term outcome in patients with acute ischemic stroke
Dandan LU ; Lan HOU ; Shuyan WEI ; Li WAN ; Xuan LI ; Xuezheng LU ; Jing WANG ; Pei WANG
International Journal of Cerebrovascular Diseases 2016;24(3):193-197
Objective To investigate the associations of baseline serum uric acid, bilirubin levels with short-term outcome in patients with acute ischemic stroke. Methods The clinical data in successive patients with acute ischemic stroke were colected, including the serum levels of uric acid and bilirubin on admission, the National Institutes of Health Stroke Scale (NIHSS) score, and the modified Rankin scale (mRS) score at discharge or at day14 (mRS 0-2 was defined as good outcome, > 2 was defined as poor outcome). Results A total of 162 patients with ischemic stroke were enroled, including 114 in the good outcome group and 48 in the poor outcome group. There were significant differences in proportions of the patients with diabetes melitus (51. 75% vs. 75. 00% ; χ2 = 7. 526, P = 0. 006), previous history of stroke or transient ischemic attack (TIA) (18. 42% vs. 50. 00% ; χ2 = 17. 790, P = 0. 001), as wel as the baseline diastolic blood pressure (87. 061 ± 12. 245 mmHg vs. 82. 375 ± 10. 949 mmHg; t = 2. 293, P = 0. 023; 1 mmHg =0. 133 kPa), high-density lipoprotein cholesterol (1. 604 ± 0. 299 mmol/L vs. 1. 265 ± 0. 206 mmol/L; t =3. 227, P = 0. 002), fasting glucose (2. 875 ± 0. 438 mmol/L vs. 8. 160 ± 0. 592 mmol/L; t = - 4. 761, P <0. 001), uric acid (289. 365 ± 77. 168 μmol/L vs. 248. 206 ± 66. 206 μmol/L; t = 3. 111, P = 0. 002), total bilirubin (14. 673 ± 2. 213 μmol/L vs. 10. 395 ± 2. 714 μmol/L; t = 3. 779, P = 0. 001 ), direct bilirubin (6. 036 ± 1. 392 μmol/L vs. 4. 956 ± 1. 379 μmol/L; t = 2. 088, P = 0. 038), and indirect bilirubin (8. 634 ± 2. 307 μmol/L vs. 5. 439 ± 1. 223 μmol/L; t = 4. 219, P < 0. 001) levels between the 2 groups. Multivariate logistic regression analysis showed that the previous history of stroke or TIA (odds ratio [ OR ] 3. 751, 95% confidence interval [CI ] 1. 395-10. 091; P = 0. 009) and baseline NIHSS score (OR 2. 723, 95% CI 1. 093-6. 783; P = 0. 031) were the independent risk factors for poor outcome of ischemic stroke; while uric acid (OR 0. 357, 95% CI 0. 141-0. 900; P = 0. 029), high-density lipoprotein (OR 0. 262, 95% CI 0. 079-0. 870; P = 0. 029), and indirect bilirubin (OR 0. 117, 95% CI 0. 025-0. 539; P = 0. 006) were independently correlated with good outcome. Conclusions The increased baseline uric acid and indirect bilirubin levels are the favorable factors for good outcome in patients with acute ischemic stroke.
4.Effect of ultraviolet A radiation on the expression and secretion of cathepsin G by human dermal fibroblasts
Qingfang XU ; Wei HOU ; Wei LAI ; Yue ZHENG ; Chen LIU ; Chun LU
Chinese Journal of Dermatology 2014;47(12):873-876
Objective To investigate the effect of ultraviolet A (UVA) radiation on the expression and secretion of cathepsin G (CatG) by human dermal fibroblasts.Methods Dermal fibroblasts were isolated from the foreskins of boys,and subjected to primary culture and subculture.After 10 or less passages,the fibroblasts were collected and divided into several groups to be irradiated with 10 J/cm2 UVA followed by 24,48 and 72 hours of additional culture,or be irradiated with 10,20 and 30 J/cm2 UVA followed by 24 hours of additional culture,with those receiving no treatment serving as the control group.Subsequently,cells and culture supernatant were collected,real time PCR and Western blot were performed to detect the expressions of CatG mRNA and protein respectively in these cells,and enzyme-linked immunosorbent assay (ELISA) was conducted to measure the expression of CatG protein in the culture supernatant of these cells.Results Compared with the control group,the fibroblasts irradiated with 10 J/cm2 UVA showed a significant increase at 24,48 and 72 hours in the expressions of CatG mRNA (0.376 ± 0.014 vs.0.183 ± 0.003,0.308 ± 0.022 vs.0.185 ± 0.005,0.296 ± 0.032 vs.0.182 ± 0.004,respectively,all P< 0.05) and protein (1.80 ± 0.12 vs.0.96 ± 0.06,1.41 ± 0.17 vs.0.95 ± 0.22,1.27 ± 0.09 vs.1.00 ± 0.14,respectively,all P < 0.05),as well as in the supernatant level of CatG protein ((161.35 ± 7.55) vs.(122.45 ± 6.46) ng/L,(141.76 ± 2.95) vs.(124.17 ± 6.15) ng/L,(139.63 ± 3.04) vs.(121.72 ± 3.17) ng/L respectively,all P <0.05),with the strongest increase observed at 24 hours.At 24 hours after 10,20 and 30 J/cm2 of UVA radiation,the expression of CatG mRNA in irradiated fibroblasts was 1.90,2.51 and 3.04 times respectively (all P < 0.05),the expression of CatG protein was 1.88,3.97 and 4.72 times respectively (P < 0.05),and the supernatant level of CatG protein was 1.36,1.50 and 1.66 times respectively (P < 0.05),that in the control group,and there was an increasing trend in all the above three parameters with increasing dose of UVA.Conclusion Acute UVA radiation can promote the expression and secretion of CatG by human dermal fibroblasts.
5.MAPK pathway regulates ultraviolet A-induced cathepsin K expression in human dermal fibroblasts
Qingfang XU ; Wei HOU ; Yue ZHENG ; Chen LIU ; Zijian GONG ; Chun LU ; Wei LAI
Chinese Journal of Dermatology 2014;47(8):543-547
Objective To investigate whether ultraviolet A UVA)-induced CatK expression is regulated by the mitogen-activated protein kinases (MAPK) signaling pathway in human dermal fibroblasts in vitro.Methods Human dermal fibroblasts were obtained from circumcised foreskin of children,and subjected to primary culture.After several passages of subculture,some fibroblasts were irradiated with UVA at a dose of 10 J/cm2.Western blot was performed to measure the expressions of total and phosphorylated JNK (t-and p-JNK) and P38 (t-and p-P38) at 0.75,1.5,3 and 6 hours after the irradiation.Some fibroblasts were divided into six groups:control group receiving no treatment,SP group treated with SP600125 of 800 nmol/L,SB group treated with SB203580 of 10 μmol/L,UVA group irradiated with UVA at a dose of 10 J/cm2,UVA-SP group treated with SP600125 for 1 hour before and for 1.5 or 48 hours after UVA irradiation at 10 J/cm2,UVA-SB group treated with SB203580 for 1 hour before and for 1.5 or 48 hours after UVA radiation at 10 J/cm2.Subsequently,Western blot was performed to determine the expressions of p-c-Jun and p-MAPKAPK2 in these groups at 1.5 hours after the UVA irradiation,and reverse transcription (RT)-PCR and Western blot to detect the mRNA and protein expressions of CatK at 48 hours after the UVA irradiation,respectively.Statistical analysis was carried out by t test,one way analysis of variance and least significant difference (LSD)-t test.Results The expression levels (gray values) of p-JNK and p-P38 were significantly increased at 0.75 hour (4.77 ± 0.19 and 2.44 ± 0.13 respectively,both P < 0.05) and 1.5 hours (4.68 ± 0.09 and 2.30 ± 0.04 respectively,both P < 0.05),but showed no significant changes at 3 hours (both P > 0.05) and 6 hours (both P > 0.05) after the UVA irradiation compared with those before the irradiation (3.2 ± 0.27 and 1.61 ± 0.08 respectively).A significant decrease was observed in the expression of p-c-Jun in the UVA-SP group and p-MAPKAPK2 in the UVA-SB group compared with the UVA group (p-c-Jun,2.55 ± 0.48 vs.4.85 ±0.96; p-MAPKAPK2,1.16 ± 0.12 vs.2.46 ± 0.09,both P < 0.05).The CatK mRNA and protein expressions were attenuated by 61.1% and 44.3% respectively in the UVA-SP group (both P < 0.05),and by 71.3% and 50.4% respectively in the UVA-SB group (both P < 0.05) in comparison with the UVA group.The UVA-SP group also showed a significant reduction in CatK mRNA and protein expressions as compared with the UVA-SB group (both P < 0.05).Conclusion Both JNK and P38 signaling pathways,especially the JNK pathway,may contribute to the upregulation of CatK expression in dermal fibroblasts induced by UVA irradiation.
6.Clinical observation of 15° face-changing acetabular cup in total hip replacement for the treatment of developmental dysplasia of hip.
Wei-kun HOU ; Lin LIU ; Chao LU ; Kan PENG ; Zhi YANG ; Ke XU ; Peng XU
China Journal of Orthopaedics and Traumatology 2016;29(6):526-529
OBJECTIVETo explore early clinical effect of acetabular cup in total hip replacement for the treatment of Crowe II developmental dysplasia of hip.
METHODSEighteen patients (18 hips) with Crowe type II developmental dysplasia of hip were treated with total hip replacement from September 2001 to July 2013. Among them,including 13 males and 5 females aged from 42 to 60 years old with an average of 47.6 years old; the courses of diseases ranged from 9 to 22 years with an average of 13.5 years. All the patients had hip joint pain, limb shortening and limited hip function before operation. Harris score of hip joint were used to evaluate recovery of function at 1 day and 12 months after operation. Prosthetic coverage of acetabular cup at 1 week after operation was observed by using radiography.
RESULTSEighteen patients (18 hips) were followed up from 12 to 24 months with an average 17 months. All incisions were healed at stage I. No deep vein thrombosis, hip dislocation, periprosthetic joint infection and prosthesis loosening were occurred. No revision surgery during follow-up period. Prosthetic coverage of acetabular cup was more than 80% at 1 week after operation. Harris score were increased from 42.67 ± 5.06 before operation to 94.79 ± 3.27 at 12 months after operation (t = -45.269, P < 0.001).
CONCLUSIONFor type Crowe II developmental dysplasia of hip patients, 15° face-changing acetabular cups in THR could obtain higher actebular component coverage rate and satisfactory early clinical effects.
Acetabulum ; surgery ; Adult ; Arthroplasty, Replacement, Hip ; instrumentation ; Female ; Follow-Up Studies ; Hip Dislocation, Congenital ; surgery ; Hip Joint ; surgery ; Hip Prosthesis ; Humans ; Male ; Middle Aged
7.Preparation of bovine serum albumin nanoparticles loaded with gemcitabine and the cytotoxic effect on pancreatic cancer cells in vitro
Jinming LI ; Wei CHEN ; Jiang LONG ; Chen JIN ; Weiyue LU ; Quanxing NI ; Deliang FU ; Huimin HOU
Chinese Journal of Pancreatology 2008;8(5):295-297
Objectives To investigate preparation of gemcitabine albumin nanoparticles, and its property of slow-release, the cytotoxic effect on pancreatic cancer cells (PANC1) in vitro, for improving the effect of regional intra-arterial infusion chemotherapy in pancreatic cancer with new medicament in the future. Methods The gemcitabine albumin nanoparticles were prepared with bovine serum albumin and gemcitabine with the desolvation-crosslink method, the concentration of gemcitabine was detected by high performance liquid chromatography (HPLC). The cytotoxic effect on pancreatic cancer cells in vitro were detected with MTT colorimetric assay. Results The mean diameter of gemcitabine albumin nanoparticles was (156.2±2.2) nm, and Zeta potential was (-20.4±1.41)mV, drug loading was 10.8%, drug release time in virto was 3 hours respectively. Gemcitabine albumin nanoparticles (0.01~50 μg/ml) had a 31%~44% inhibitory rate on PANC1 cell, which was similar to the inhibitory rate of same concentration of gemcitabine (26%~47%). Conclusions The new preparation of gemcitabine albumin nanoparticles had obvious drug slow-release effect, which may help improve the effect of regional intra-arterial infusion chemotherapy for pancreatic cancer.
8.X-ray induced TCR gene mutation of peripheral blood lymphocytes
Dianjun HOU ; Ya MA ; Wei LIU ; Jianwei QIAO ; Feng LU ; Ximei SHANG ; Jieqing LI ; Li FENG
Chinese Journal of Radiological Medicine and Protection 2009;29(2):151-154
objective To study the TCR gene mutation in peripheral blood lymphocytes induced by X-ray exposure using cultured lymphocytes cloning method.Methods Freshly isolated peripheral lymphocytes from healthy aduh donors were irradiated with X-ray in doses ranging from 0 to 8 Gy and cultured with interleukin2 and phytohemagglutinin for 7 days.The mutant frequencies of TCR gene(TCR MF)were detected by flow cytonletry and the dose response curves were fitted.Results TCR MF increased with the dose going up.An aquadratic polynomial dose response model was fitted.Conclusions TCR gene mutation could which serve as a potential biological dosimeter.It might be applied for the estimation of biological dose in emergency exposure.
9.Consistency of T-cell receptor gene mutation between lymphocytes in rats after irradiation in vivo and in vitro
Yan SHI ; Wei LIU ; Dianjun HOU ; Ya MA ; Jianwei QIAO ; Feng LU ; Ximei SHANG
Chinese Journal of Radiological Medicine and Protection 2010;30(5):561-563
Objective To observe the consistency of T-cell receptor (TCR) genes mutation in lymphocytes in rats after irradiation in vivo and in vitro.Methods A total of 48 female rats were randomly divided into 6 equal groups.Peripheral blood samples from them were collected to separate the lymphocytes and then irradiated to X-ray irradiation with the dose rate of 200 cGy/min at the doses of 0,0.5,0.75,1.0,2.0,and 3.0 Gy,respectively.Then all the lymphocyte samples were cultured for 7 days.Flow cytometry with direct immunofluorescence was used to detect the TCR gene mutation.The levels of TCR gene mutant frequency (TCRMF) of different groups were calculated.Results The TCRMF levels of different groups after irradiation in vivo and in vitro all displayed a dose-dependent manner and there were no significant differences in the TCRMF between different dose irradiation groups(t = -1.1-0.3 ,P >0.05).Conclusions A consistency of TCRMF after irradiation in vivo and in vitro is proven.The results of TCRMF of peripheral blood lymphocytes irradiated in vitro by flow cytometry can precisely reflect the TCR genes mutation after whole-body irradiation.
10.Cloning, expression and purification of Nocardia brasiliensis proteion P61 with biological activity
Xingzhao JI ; Lu TANG ; Xuexin HOU ; Lina SUN ; Chao WEI ; Shuai XU ; Chenchen SI ; Zhenjun LI
Chinese Journal of Zoonoses 2017;33(3):260-263
We constructed prokaryotic recombinant expression vector of P61 gene from Nocardia brasiliensis,expressing P61 protein with biological activity in E.coli,and lay a foundation for further studies related to P61.P61 gene was synthesized and cloned into an expression vector pET-30a(+).The recombinant vector was transformed into Escherichia coli BL21 and induced with IPTG.The production was analyzed with Western blot and the catalase activity of P61 was tested with Catalase Assay Kit.The protein of P61was successfully expressed in E.coli with solubility and high catalase activity,and could be identified by anti-N.brasiliensis sera from mice.The prokaryotic expression plasmid of protein P61 was constructed successfully and can be expressed efficiently in E.coli BL21 cells with higher catalase.