1.Effects of tobacco extract on the proliferation and HSP70 expression of human periodontal ligament cells
Wei LI ; Dong CHEN ; Kunyang LI ; Yun FAN ; Fangfang YU
Journal of Practical Stomatology 2014;(2):211-214
Objective:To study the effects of smokeless tobacco extract(ST)on the proliferation and the heat shock protein 70 (HSP70)expression of human periodontal ligament cells(hPDLCs).Methods:hPDLCs were cultured in vitro and identified by im-munohistochemistry(IHC).The cells were stimulated with ST at 0.01 6 -50 g/L respectively for 24 h,the proliferation was examine by MTT assay,HSP70 expression was detected by immunehistochemical staining and Western Blot.Results:ST inhibited the prolifer-ation and increased HSP70 expression in cytoplasm and nucleus at 0.4 -50 g/L dose dependantly.Conclusion:ST may inhibit the proliferation and increase HSP70 expression of hPDLCs in a dose depandant manner.
2.Effects of anthraquinones from Cassia obtusifolia L. on cholesterol biosynthesis in cells
Chuhua LI ; Xue LI ; Kunyang FANG ; Baoliang GUO
Chinese Journal of Tissue Engineering Research 2008;12(33):6593-6596
BACKGROUND: The effective chemical constituents and the precise action mechanism of Cassia obtusifolia L. seed to rats with hypercholesterolemia are not clear. OBJECTIVE: To investigate effects of anthraquinones from Cassia obtusifolia L. on hypolipidemic activity and endogenous cholesterol biosynthesis in rats with experimental hyperlipoidemia, and to explore the effective compound and the mechanism of Cassia obtusifolia L. seed on hypolipidemic action. DESIGN, TIME AND SETTING: The randomized controlled animal experiment was performed from September 2003 to May 2004. All rats were raised and tested at the College of Life Sciences, South China Normal University. Blood sample was collected from the tail vein at the Institute of Biotechnology to detect blood lipid, survival rate, and to culture cells. MATERIALS: Forty-five male Sprague Dawley (SD) rats were used to establish animal models of hypedipoidemia by intragastrically with fat emulsion. Cassia obtusifolia L. seed was purchased from Guangzhou Dispensary, China, and further identified by South China Plant Institute. Anthraquinones were extracted from Cassia obtusifolia L. by the Institute of Biotechnology of South China Normal University. METHODS: Forty-five male SD rats were randomly divided into three groups (n=15): a control group, 80 mg/kg and 20 mg/kg anthraquinones groups. Rat models in each group were given fat emulsion in the morning and afternoon 2 days after model induction, once in morning. Rat modes were treated with corresponding doses of drugs in the two experimental groups. Rat models were administrated with the same volume of saline in the control group, once a day, for 20 days. MAIN OUTCOME MEASURES: Effect of anthraquinones on endogenous cholesterin in Chinese hamster oocytes was measured by amphotericin B cell models. The levels of serum total cholesterol, triglyceride, low-density iipoprotein-cholesterol (LDL-C) and high-density lipoprotein-cholesterol (HDL-C) were detected by enzyme endpoint method. Survival rate of Chinese hamster oocytes (A570) was tested by methyl thiazolyi tetrazolium (MTT) with spectrophotometric determination. RESULTS: Anthraquinones significantly reduced total cholesterol, triglyceride, LDL-C levels and increased HDL-C levels in hyperlipidemic rats in a dose-dependant pattern. Anthraquinones elevated the survival rate of Chinese hamster oocytes. CONCLUSION: Anthraquinones can decrease blood lipid levels. Inhibition of cholesterol synthesis of anthraquinones may be one of the underlying mechanism involved in decreasing blood lipid.
3.Circulating endothelial cells participate in the in vivo endothelialization of vascular prosthesis: An animal experiment
Yi WANG ; Yiren CHEN ; Kunyang DAI ; Hongwen NIU ; Bo WU ; Li LI ; Dachuan QI
Chinese Journal of Tissue Engineering Research 2007;11(50):10209-10212
BACKGROUND: Experiments have demonstrated that autologous vascular endothelial cells if transplanted onto artificial vascular cavosurface, can enhance the patency rate of vasotransplantation. Whether seeding of prostheses interposition grafts with bone marrow-derived endothelial cells is effective for in vivo endothelialization of artificial vessels remains unclear.OBJECTIVE: To observe the effect of endothelialization of vascular prosthesis by seeding prostheses interposition grafts with bone marrow-derived endothelial cells in animals.DESIGN: A controlled animal experimental study.SETTING: Shanghai Sixth People's Hospital.MATERIALS: This study was carried out in the Shanghai Sixth People's Hospital between September 2000 and October 2001. Twenty hybrid dogs from Shanghai, of either gender, aged 1.0 to 2.0 years old, weighing (18.7±2.3) kg, were involved in this study.METHODS: Bone marrow-derived mononuclear cells were isolated from the dogs. The endothelialization of ePTFE prostheses interposition grafts (4 mm×4 cm and 8 mm×5 cm)was carried out. Common carotid artery transplantation:Ten laboratory dogs were involved. Common carotid artery of 4 cm was resected from each dog. ePTFE prostheses interposition grafts of 4 mm×4 cm was transplanted into the bilateral common carotid artery, and prostheses interposition grafts were performed endothelialization, namely experimental group. Those prostheses interposition grafts, which were not performed endothelialization, were named as control group. Five dogs were used in each group. Patency rate and blood flow rate of transplanted vessels were detected with a color ultrasonograph 2 weeks and 2 months after operation.Inferior caval vein transplantation: Six of the rest 10 dogs were used for experiments. Under the anesthesia, 8-10 cm inferior caval vein was dissociated from each dog. Its two ends were blocked, and about 5 cm inferior caval vein was resected. ePTFE endothelialized vascular prosthesis with 8 mm in diameter and 5 cm in length was anastomosed end to end with 5-0 Prolene. The other 4 dogs were used for control experiment. ePTFE vascular prosthesis with the same specification was used as prostheses interposition graft. Vascular patency rate was determined 2 months after operation.At the same time, coverage rate and intimal thickness of transplanted vascular endothelial cells and vascular intimal thickness were determined.MAIN OUTCOME MEASURES: ①The patency rate and blood flow rate of transplanted vessels at different time points. ②Coverage rate of transplanted vascular endothelial cells and vascular intimal thickness.RESULTS:① At 2 weeks and 2 months after common carotid artery transplantation, the patency rate of experimentalside was 100%(5/5)and 60%(3/5), respectively, and that of control side was 40%(2/5)and 0%(0/5), respectively. At postoperative 2 months, the mean blood flow rate in the experimental group was obviously smaller than that in the control group (P < 0.05). At 2 months after inferior caval vein transplantation, the patency rate of experimental group and control group was 83%(5/6)and 50%(2/4), respectively. ②At 2 weeks after common carotid artery transplantation and inferior caval vein transplantation, the coverage rate of vascular endothelial cells in the experimental group was significantly larger than that in the control group, separately (P < 0.05). At 2 months after each transplantation, the vascular intimal thickness in the experimental group was significantly smaller than that in the control group (P < 0.05).CONCLUSION: Seeding of ePTFE prostheses interposition grafts with bone marrow-derived endothelial cells can rapidly accomplish in vivo endothelialization and inhibit intimal hyperplasy; Circulating endothelial cells, as the potential source of endothelial cells, have certain clinical application values.
4.Effect of Progressive Resistance Training on Upper Limb Function in Breast Cancer Patients
He LI ; Yixiao WANG ; Tieying SHI ; Qigui LIU ; Chunli SONG ; Kunyang LI
Chinese Journal of Rehabilitation Theory and Practice 2015;(11):1356-1359
Objective To explore the effect of postoperative progressive resistance training (PRT) on the upper limb function in breast cancer patients. Methods 66 breast cancer patients were randomly divided into intervention group (n=33) and control group (n=33). The intervention group accepted a 12-week PRT and the control group accepted the routine training since 4-6 weeks postoperative as they admitted to hospital for their first chemotherapy. All the patients received a same content of health education. Results The grip strength, range of motion of abduction and flexion of shoulder were more in the intervention group than in the control group after intervention (P<0.01). Conclusion PRT can effectively improve the function of upper limbs in breast cancer patients.
5.Effects of Dengzhanhua Capsule on Kidney Tissue Inflammatory Cytokines in Chronic Renal Failure Rats
Yibin DU ; Xiao WU ; Feng WU ; Qi LI ; Kunyang ZHANG ; Yanrui DUAN ; Chuqiao XIE ; Liqun HE
Chinese Journal of Information on Traditional Chinese Medicine 2014;(12):63-65
Objective To observe the effects of Dengzhanhua Capsule on kidney tissue inflammatory cytokines in chronic renal failure rats;To explore its possible mechanism for the efficacy in chronic renal failure. Methods Sixty SD rats were randomly divided into normal control group, model group, benazepril group and Dengzhanhua group, 15 rats in each group. Chronic renal failure rat model was established by Platt 5/6 nephrectomized. Benazepril (0.29 mg/100 g) was given to rats in the benazepril group by gastrogavage. Dengzhanhua Capsule (0.3 g/100 g) was given to rats in the Dengzhanhua group by gastrogavage. Normal saline was given to rats in the normal group and the model group by gastrogavage. The whole treatment period was twelve weeks. Expressions of TGF-β1 and PAI-1 were determined by semi-quantitative RT-PCR after treatment. Concentrations of kidney tissue inflammatory cytokines IL-6 and TNF-α were determined by ELISA. Results Expressions of TGF-β, PAI-1 and IL-6, TNF-αin benazepril group and Dengzhanhua group were significantly lower than those in model group (P<0.05). Compared with benazepril group, it was significantly lower in Dengzhanhua group (P<0.05). Conclusion Dengzhanhua Capsule can reduce kidney tissue inflammatory in chronic renal failure rats, and inhibit renal fibrosis.
6. 2-Chloroethanol enhanced the expression of AQP4 protein in astrocytes via NMDAR/cAMP/PKA signaling pathway
Chaohong LUO ; Jingjing GUO ; Tiansheng XU ; Kunyang LI ; Yaping JIN
China Occupational Medicine 2019;46(01):15-21
OBJECTIVE: To investigate the role of N-methyl-D-aspartate receptor(NMDAR)/cyclic adenosine monophosphate(cAMP)/protein kinase A(PKA) signaling pathways in regulating 2-chloroethanol-induced aquaporin-4(AQP4) expression in astrocytes(AS). METHODS: i) AS in logarithmic growth phase were treated with 2-chloroethanol at the doses of 0.0, 7.5, 15.0 and 30.0 mmol/L for 12 hours, and the cells were collected for detection. ii) The AS in logarithmic growth phase were divided into blank control group, inhibitor control group, 2-chloroethanol group, and inhibitor intervention group. The inhibitor included dizocilpine(MK-801) and N-(2-[p-bromocinnamylamino-]ethyl)-5-isoquinolinesulfonamide(H89). The blank control group did not receive any treatment. The inhibitor control group was treated with a concentration of 10.0 μmmol/L MK-801 or 15.0 μmmol/L H89. The MK-801 intervention group was pretreated with MK-801 at a concentration of 10.0 μmmol/L for 30 minutes. The H89 intervention group was pretreated with H89 at a concentration of 15.0 μmmol/L for 1 hour. After the intervention, the AS in 2-chloroethanol group and MK-801, H89 intervention group were stimulated with 2-chloroethanol at a dose of 30.0 mmol/L for 12 hours. iii) The AS in each group were collected and used for Western blotting and real-time fluorescence quantitative polymerase chain reaction analysis to detect the protein and mRNA expression of AQP4, NMDAR receptor main subunit(NR1), NMDAR receptor 2 B subunit(NR2 B) and calmodulin dependent protein kinaseⅡ(CaMKⅡ). The Western blotting was adopted to detect the expression of phosphorylase-CaMKⅡ(p-CaMKⅡ) and PKA. Colorimetric method was used to detect the concentration of calcium(Ca~(2+)) in AS. The enzyme-linked adsorption test was used to measure adenylate cyclase(AC) activity and cAMP levels. RESULTS: i) The relative expression of protein and mRNA of AQP4, NR1 and NR2 B, PKA at protein level and CaMKⅡ at mRNA level, and the ratio of p-CaMKⅡ/CaMKⅡ protein, the concentration of Ca~(2+), AC activity and cAMP level in 30.0 mmol/L group were higher then those of 0.0 mmol/L group in AS(P<0.05). The relative protein expression of PKA and the concentration of Ca~(2+) increased with the increase of 2-chloroethanol(P<0.05). ii) The relative protein expression of AQP4 and the concentration of Ca~(2+) in the 2-chloroethanol group were higher than that of the blank control group and MK-801 control group(P<0.05). The relative protein expression of AQP4 and the concentration of Ca~(2+) in MK-801 intervention group were lower than that in 2-chloroethanol group(P<0.05). The relative protein expression of AQP4 and PKA in 2-chloroethanol group were higher than that of the blank control group and H89 control group(P<0.05). The relative protein expression of AQP4 and PKA in H89 intervention group was lower than that in 2-chloroethanol group(P<0.05). CONCLUSION: The 2-chloroethanol timulation induces the expression of AQP4 by activating NMDAR/cAMP/PKA signaling pathway in AS.