1.Effects of gypenosides on the expressions of p53 and p21 protein in photodamaged BALB/c mouse skin
Limei YUAN ; Danqi DENG ; Yang LI ; Liu LIU
Chinese Journal of Dermatology 2011;44(4):253-255
Objective To explore the mechanism underlying the effects of gypenosides (GP) against photodamage. Methods Eighty BALB/c mice were equally divided into 8 groups, i.e., blank control group (untreated), UVB model group (irradiated with UVB), GP I group (irradiated with UVB before topical application of GP), GPⅡ group (irradiated with UVB followed by topical application of GP), VitE I group (irradiated with UVB after topical application of Vitamine E cream), VitE Ⅱ group (irradiated with UVB followed by topical application of Vitamine E cream), Vehicle group Ⅰ (irradiated with UVB after application of the drug vehicle),and Vehicle group Ⅱ (irradiated with UVB before application of the drug vehicle). UVB irradiation was performed once every other day for 14 days. Mice were sacrificed after the last irradiation and skin specimens were obtained from the irradiated sites, and the levels of p53 and p21 protein were measured by Western blot in the specimens. Results The expression level of p53 protein was significantly lower in the blank control group than in the UVB model group (0.11 ± 0.08 vs. 0.22 ± 0.12) and GP Ⅰ group (0.44 ± 0.23, P < 0.01),in the blank control group and UVB model group than in the GP Ⅱ group (0.48 ± 0.24, P < 0.01, 0.05). VitE Ⅰ group (0.49 ± 0.29) and VitE II group (0.50 ± 0.27) were similar to the GP groups in the expression of p53 protein. No statistical difference was observed in the expression of p21 protein between the eight groups. Conclusion The upregulation of p53 protein expression in epidermal cells may be related to the mechanisms underlying the protective effect of 1.5% GP cream against photodamage.
2.Effects of gypenosides on nuclear factor κB and p38 mitogen activated protein kinase signaling pathways in photodamaged skin of mice
Danqi DENG ; Yang LI ; Yilin WANG ; Limei YUAN ; Liu LIU
Chinese Journal of Dermatology 2012;45(3):195-198
Objective To observe the effects of gypenosides (GP) on nuclear factor κB (NF-κB) and p38 mitogen activated protein kinase (p38MAPK) signaling pathways in photodamaged skin of mice,and to explore the mechanisms underlying the protective effects of GP against photodamage.Methods Eighty Balb/C female mice were randomly divided into 8 groups: blank control group receiving no treatment,ultraviolet B (UVB) model group receiving UVB irradiation for 60 seconds,GP group Ⅰ receiving topical GP treatment followed by UVB irradiation,GP group Ⅱ receiving UVB irradiation followed by topical GP treatment,VitE group Ⅰ receiving topical VitE treatment followed by UVB irradiation,VitE group Ⅱ receiving UVB irradiation followed by topical VitE treatment,matrix group Ⅰ receiving topical matrix treatment followed by UVB irradiation,matrix group Ⅱ receiving UVB irradiation followed by topical matrix treatment.UVB irradiation lasted 60 seconds at one time,and was given once every other day for 7 times to establish a skin model of photodamage.The interval between irradiation and topical treatment was 30 minutes in all the groups except the control group and UVB model group.After the last treatment,mice were sacrificed.Western blot was performed to measure the protein expressions of inhibitor of NF-κB(IκB),inhibitor of NF-κB kinase (IKK),p38MAPK as well as phosphorylated p38MAPK (pp38) in skin tissue from the mice.Results No expressions of IκB or IKK were observed in the blank control group.The expression level of IκB was 0.40 ± 0.07 in UVB model group,significantly lower than that in GP group Ⅰ (1.63 ± 0.85,P < 0.05) and GP group Ⅱ (0.90 ± 0.40,P < 0.05),whereas the level of IKK protein was higher in UVB model group than in the GP group Ⅰ and Ⅱ (2.01 ± 1.75vs.0.23 ± 0.12 and 0.45 ± 0.29,both P < 0.05).No significant difference was observed in the expression of IκB or IKK proteins between the GP group Ⅰ,Ⅱ,VitE group Ⅰ and Ⅱ or in the expression of p38MAPK between the 8 groups.The phosphorylated p38MAPK expression in UVB model group was significantly higher than that in GP group Ⅰ and Ⅱ (0.835 ± 0.049 vs.0.425 ± 0.054 and 0.571 ± 0.090,both P< 0.05),but similar to that in VitE groups.Conclusions UVB can activate NF-κB and phosphorylated p38MAPK signaling pathways; GP 1.5% cream can inhibit UVB-induced activation of NF-κB and p38MAPK pathways,which may be one of the mechanisms underlying its protective effects against inflammation and photodamage.
3.Culture,proliferation and differentiation of human marrow mesenchymal stem cells to cardiomyocytes in vitro
Fangge DENG ; Xiuying ZHANG ; Limei QU ; Yulin LI
Chinese Journal of Immunology 1985;0(02):-
95% purity were treated for 24 h with 1,5,10 and 20 ?mol/L 5-azacytidine to induce cellular differentiation.Expression of the cardiac-specific marker-troponin Ⅰ and desmin,identified by immunohistochemistry,was used to identify cardiac muscle differentiation.Results:hMSCs expressed a high level of CD44(hMSCs 93.26%?2.48% vs 3.42%?1.09% in a control group,P
4.Isolation and identification of hepatic stem cells from rat fetal liver in vitro
Limei QU ; Xiuying ZHANG ; Xinrui WANG ; Fangge DENG ; Yulin LI
Journal of Jilin University(Medicine Edition) 2006;0(03):-
Objective To isolate and cultivate hepatic stem cells(HSCs) from rat fetal liver in vitro and identify their biological features.Methods Collagenase perfusion method and mechanical cutting method were used to isolate HSCs from rat fetal liver which were then cultivated by H-DMEM containing 10% fetal bovine serum.The cell surface antigen expression of HSCs was observed with immunocytochemical method under confocal laser scanning microscope.Results The isolated HSCs from rat fetal liver grew to monolayer 5 d after cultivation in vitro.They presented pykno-round cells and distinct borderline under the light microscope.After 8 d the cells grew like epithelium.These cells expressed AFP antigen,CK 18 and CK19.Conclusion The cultivated cells are proved to be HSCs and can proliferate quickly in vitro.
5.Observation of Therapeutic Effect of Tacrolimus Ointment and Mometasone Furoate Cream in the Treatment of Chronic Actinic Dermatitis
Juan CHEN ; Danqi DENG ; Limei YUAN ; Hanfei HU ; Zhaoxia HAN
Journal of Kunming Medical University 2014;(1):59-62
Objective To evaluate the efficacy of 0.1% tacrolimus ointment and 0.1% mometasone furoate cream in the treatment of chronic actinic dermatitis (CAD). Methods Forty male patients with CAD were recruited and divided into two groups randomly.Twenty cases were treated with 0.1%tacrolimus ointment (Group A), and the other 20 cases were treated with 0.1% mometasone furoate cream (Group B) . The medications mentioned were applied topically to the lesions on the face twice a day and mizolastine tablet 10 mg per day given orally for 4 weeks. The therapeutic efficacy and side effects of medications were observed. The enzyme linked immunosorbent assay (ELISA) method was used to measure the serum levels of IFN-γ, IL-2 and IL12 in CAD patients before and after treatment with topical tacrolimus ointment and mometasone furoate cream. Results (1) Both groups had overall response rates of 100%, with no statistically significant difference ( > 0.05) . (2) Serum levels of IFN-γ,IL-2 and IL-12 were down-regulated after treatment in both treatment groups respectively ( < 0.01) . No statistically significant difference was found between the two treatment groups ( > 0.05) . Conclusion 0.1%tacrolimus ointment is effective in the treatment of CAD. Its therapeutic efficacy is equivalent to that of 0.1%mometasone furoate cream. It can be used as a possible steroid sparing equivalent.
6.A survey of antibiotic application for hospitalized patients receiving oral and maxilla facial surgery
Zhiqing LIU ; Limei DENG ; Min CHEN ; Yuenian WANG
Journal of Practical Stomatology 1995;0(04):-
Objective:To investigate the situation for clinical antibiotic usage in West China Hospital of Stomatology. Methods:By using specially designed survey table, a retrospective study was carried out and the data of antibiotic usage for all the patients from March to August 2003 were analyzed by computer.Results:A total of 1139 cases were investigated and antibiotics were used in 84.72% of the cases. Single drug,2-drug-combination and 3 or more were used in 36.52%,43.91% and 4.30% of the patients.Antibiotics were administrated by intravenous injection in 93.7% of the cases.The antibiotic application for preventive purpose was in 64.87% of all the subjects.The cost of antibiotics contributed to 48.78% in total drug expenses.Conclusion:It is necessary to strengthen the management of antibiotic application.
7.The expressions of notch genes in human keloid-derived mesenchymal-like stem cells.
Chengliang DENG ; Bo WANG ; Ziyang ZHANG ; Guangfeng SUN ; Jingjing ZHU ; Dali WANG ; Limei YU
Chinese Journal of Plastic Surgery 2014;30(3):197-202
OBJECTIVETo study the expressions of Notch1-4 gene in human keloid-derived mesenchymal-like stem cells, and to explore the Notch signaling pathway's role in the formation of keloid.
METHODSKeloid samples were collected to harvest human keloid-derived mesenchymal-like stem cells through two-step enzymatic dissociation method. By flow cytometry, cell phenotype of primary and P3 generation were analyzed. By immunocytochemistry, the expressions of Oct4, vimentin and CK19 were examined. Keloid-derived mesenchymal-like stem cells were induced into osteoblasts in vitro and calcium deposition was detected by Alizarin red S stain. Realtime polymerase chain reaction (RT-PCR) was used to detect the expressions of Notch1-4 mRNA in keloid-derived mesenchymal-like stem cells.
RESULTSFlow cytometry showed that keloid-derived mesenchymal-like stem cells of primary and P3 generation highly expressed CD29, CD44, CD90 from the typical MSC phenotype marker, but they failed to express HSC phenotype markers, such as CD34 and CD45. The results of immunocytochemistry showed that Oct4 from pluripotent stem cell markers and vimentin from mesenchymal cell markers was positive and CK19 from epithelial cell markers was negative. After induced differentiation into osteoblasts in vitro after 21 day, calcium nodules could be seen clearly; Notch1-4 gene were expressed in keloid-derived mesenchymal-like stem cells through RT-PCR. The relative quantitative of Notch2, Notch3 gene were higher than Notch1, Notch4 gene (P < 0.05).
CONCLUSIONSThe expression difference of different subtypes from Notch gene in human keloid-derived mesenchymal-like stem ceils may be related to self-renewal, proliferation, differentiation, and participate in the formation of keloid.
Adolescent ; Cells, Cultured ; Child ; Female ; Humans ; Keloid ; metabolism ; pathology ; Male ; Mesenchymal Stromal Cells ; metabolism ; Receptors, Notch ; metabolism
8.Association between plasma visfatin and intima-media thickness of carotid artery in patients with type 2 diabetes
Lihua GAO ; Zhansheng ZHAO ; Hui TIAN ; Bingjie WU ; Limei JIA ; Mian WANG ; Yonggui DENG ; Hong ZHOU ; Lihui ZHANG ; Shengou SU
Chinese Journal of Endocrinology and Metabolism 2010;26(1):15-16
Seventy-three patients with type 2 diabetes mellitus were divided into atheroselerosis(AS) group and non-AS group according to the intima-media thickness(IMT)of the carotid artery.The plasma visfatin level in AS group was higher than that in non-As group[(44.95±10.14 vs 34.52±9.08)μg/L,P<0.05],and both of them were higher than that of the control [(24.46±7.18)μg/L,both P<0.05 ].The visfatin level Was positively correlated with IMT,waist-to-hip ratio,visceral fat thickness,fasting insulin,and HOMA insulin resistance index.Age,duration of diabetes,HbA_(1C),and visfatin level were the major risk factors for IMT of the carotid artery.
9.Study on enterovirus infection in family close contacts of patients with hand, foot and mouth disease and the contamination of environment
Limei SUN ; Jinyan LIN ; Hanri ZENG ; Min KANG ; Huanying ZHENG ; Aiping DENG ; Jianfen HE ; Changwen KE ; Yonghui ZHANG
Chinese Journal of Infectious Diseases 2012;30(6):339-342
Objective To understand the enterovirus infection in family close contacts of patients with hand,foot and mouth disease (HFMD) and the contamination of environment.Methods Forty-one HFMD cases confirmed by laboratory from web-based surveillance system during July to August 2010 in Guangdong Province were selected.All members of the cases′ family were investigated by collecting their information on demography,habit of domestic hygiene and hygiene status in household.The stool samples of all members and the smear samples from the surface of family belongings from 16 families were collected and the enterovirus was detected by real time quantitative polymerase chain reaction.The data were analyzed by chi square teat and t test.ResultsForty-one HFMD cases′ families and 135 close contacts were included in this survey.The infection rate of the enterovirus was 39.2% (53/135) in all close contacts.Of all the investigated families,the infectionrate was 58.5% (24/41) in family with one or more close contacts and 9.8% (4/41) in family with all close contacts.The differences of infection rates of enterovirus among the members of parents (32.5%,25/77),grandparents/aunts/ uncles (43.3%,13/30) and cousins (53.6%,15/28) didn′t show statistical significance (χ2 =4.07,P=0.131).The infection rate of enterovirus in close contacts from family with more than 5 members was higher than that from family with 4 or less members (OR=1.4,95%CI 1.1-1.9).Among 135 close contacts,27.4% (37/135) were infected with the same types of entervirus as that of HFMD case in the family and 11.9% (16/135) were infected with the different virus types.In 33 family belongings samples from 16 families,the positive rate of enterovirus detection was 6.1% (2/33).Between 17 families with enterovirus testing negative and 23 families with enterovirus testing positive in close contacts,there were no statistical differences of the family hygiene status,hand-washing of babysitter,disinfection of tableware and drinking,sharing towels,airing bedding articles and toy cleaning (P>0.05).ConclusionsThe infection rate of enterovirus in close contacts of HFMD cases is high and the enterovirus contamination exists in case family environment.Management of close contacts of HFMD cases and disinfection of the family environment are important in HFMD controls.
10.Ca(2+)-activated K(+) channel switching in smooth muscle participates in atherosclerosis development in diabetic rats.
Yan WANG ; Limei ZHAO ; Xingli SU ; Wei YU ; Xiuling DENG
Journal of Southern Medical University 2014;34(2):188-192
OBJECTIVETo investigate the changes in aorta morphology and Ca(2+)-activated K(+) (KCa) channel expression in the diabetic rats.
METHODSA diabetic rat model was established by a single intraperitoneal injection of streptozotocin (30 mg/kg) after a modified high fat and glucose diet for 8 weeks. Pathological changes in the aorta were observed with HE staining, elastic fiber staining, Masson's trichrome staining and immunohistochemistry. Both the mRNA and protein levels of KCa channels in the aorta were measured by RT-PCR and Western blotting.
RESULTSEarly atherosclerotic changes were observed in the aorta wall of the diabetic rats. The mRNA and protein levels of KCa1.1 channel α- and β-subunits were significantly decreased, while the expression of KCa3.1 channels was obviously enhanced in the middle layer of the aorta in the diabetic rats.
CONCLUSIONKCa channel switching in smooth muscles may play a role in the development of atherosclerosis in diabetic rats.
Animals ; Aorta ; pathology ; Atherosclerosis ; pathology ; Diabetes Mellitus, Experimental ; pathology ; Intermediate-Conductance Calcium-Activated Potassium Channels ; metabolism ; Large-Conductance Calcium-Activated Potassium Channel alpha Subunits ; metabolism ; Male ; Muscle, Smooth, Vascular ; metabolism ; Rats ; Rats, Sprague-Dawley