1.Inducing human bone marrow mesenchymal stem cells to differentiate into endothelial cells with indirect co-culture
Jun LU ; Wei HUANG ; Xinxin CHEN
Chinese Journal of Tissue Engineering Research 2007;0(38):-
BACKGROUND: Recent studies indicate that after indirect co-culture of neonate rat myocardial cells and bone marrow mesenchymal stem cells, bone marrow mesenchymal stem cells can differentiate into myocardial cells successfully. OBJECTIVE: To induce human bone marrow mesenchymal stem cells (hBMSCs) to differentiate into endothelial cells using human umbilical vein endothelial cells by indirect co-culture. DESIGN, TIME AND SETTING: In vitro study of cell engineering was done at the Medical Research Center of Guangdong Provincial People’s Hospital between January and July 2007. MATERIALS: Small quantities of bone marrow were obtained from 11 children with congenital heart disease but without hematologic diseases through manubrium of sternum puncture in the congenital heart defect corrective surgery after the permission of family member. Umbilical cord of full-term normal delivery healthy newborn was provided by the Department of Obstetrics, the First Affiliated Hospital of Sun Yat-sen University. Fresh cattle jugular vein was provided by Guangdong Dali Meat Cattle Butchery. METHODS: The hBMSCs were isolated and purified using density gradient centrifugation method and were cultured in vitro. Human umbilical vein endothelial cells were obtained from newborn umbilical cord by enzyme digestion. Cell culture insert with semipermeable membrane combined with 6-well plate was used to do indirect co-culture induction. Human umbilical vein endothelial cells were expanded in the cell culture insert, passage 3 bone marrow mesenchymal stem cells were expanded in the 6-well plate outside the culture insert at the density of 1?105 cells/well, the initial ratio of the two kinds of cells was 1:5, then low-glucose DMEM culture solution containing 10% fetal bovine serum was added, cells were cultured for 14 days. Co-culture of bone marrow mesenchymal stem cells and bone marrow mesenchymal stem cells was used as control. Introduced endothelioid cells were cultured and then seeded on the cell-free cattle jugular vein intravascular stent. MAIN OUTCOME MEASURES: Morphology changes of induced cells; introduced endothelioid cell surface antigen detected through immunocytochemical staining; the growth and adhesion condition of endothelioid cells on the intravascular stent observed under scanning electronic microsope. RESULTS: The morphologies of introduced endothelial cells were uniform, introduced endothelial cells presented a cobblestone-like appearance, they amplified fast and expressed endothelial cell-specific surface marker CD31 and vWF and the positive rate was over 99%. They also could form a continuous unicellular layer on the cell-free cattle jugular vein intravascular stent. CONCLUSION: Human umbilical vein endothelial cells can induce hBMSCs to differentiate into endothelial cells successfully and to adhere and grew on the cell-free cattle jugular vein intravascular stent through indirect co-culture method.
2.Clinical Study on Fluticasone Furoate Nasal Spray in the Treatment of Allergic Rhinitis
Chen YANG ; Xinxin WEI ; Zhongpu YIN
China Pharmacy 2017;28(11):1503-1506
OBJECTIVE:To investigate clinical efficacy and safety of Fluticasone furoate nasal spray in the treatment of aller-gic rhinitis(AR). METHODS:100 AR patients in our hospital during Jan. 2014-Dec. 2015 were divided into control group and ob-servation group according to random number table,with 50 cases in each group. Both groups received decongestant Oxymetazoline hydrochloride spray. Control group was additionally given Budesonide spray,once each nostril on the first day,twice each nostril on the second day,increasing suitably according to disease condition,less than 6 times a day for patients younger than 14 years old,less than 8 times a day for patients older than 14 years old;observation group was additionally given Fluticasone furoate nasal spray,once each nostril,qd for patients younger than 14 years old,once each nostril,while morning and night for patients older than 14 years old. Both groups received treatment for consecutive 30 d. The total nasal symptoms score(TNSS),total eye symp-toms score (TESS),total nasal resistance (TNR) under 75,150 Pa,nasal minimum cross-sectional area (MCA) and the occur-rence of ADR were compared between 2 groups before and after treatment. RESULTS:There was no statistical significance in above indexes between 2 groups before treatment (P>0.05). After treatment,TNSS of 2 groups and TESS of observation group were decreased significantly compared to before treatment,and TESS of observation group was significantly lower than that of con-trol group,with statistical significance (P<0.05). There was no statistical significance in TESS of control group before and after treatment(P>0.05). TNR of 2 groups under 75,150 Pa were significantly lower than before treatment,and the observation group was lower than the control group,with statistical significance (P<0.05). MCA of 2 groups were significantly lower than before treatment,without statistical significance between 2 groups(P>0.05). The incidence of ADR in observation group was significant-ly lower than control group,with statistical significance(P<0.05). CONCLUSIONS:Fluticasone furoate is similar to budesonide in the treatment of nasal symptoms of AR patients,but it is better than budesonide in improving eye symptoms and nasal ventila-tion function with milder ADR.
3.Diagnosis and treatment of poorly differentiated small intestinal neuroendocrine carcinoma
Hao WANG ; Ping CHEN ; Wei ZHAO ; Lei SHI ; Xinxin LIU
Chinese Journal of Digestive Surgery 2013;12(10):804-806
Small intestinal neuroendocrine carcinoma is poorly differentiated,highly malignant,and originated from the diffuse neuroendocrine system.It diffusely expresses the general neuroendocrine differentiation markers.The disease can be manifested as carcinoid syndrome,but most of the patients were with occult onset and absence of specific clinical symptoms,which led to difficult diagnosis before operation.On June 18,2012,a patient with manifestation of recurrent vague abdominal pain received capsule endoscopy at the Northern Jiangsu People's Hospital.The capsule endoscope was retained in the distal small intestine,and malignant ileum lesion was revealed during the surgical exploration.Finally,poorly differentiated neuroendocrine carcinoma with abdominal wall metastases was identified by pathological examination.
4.Effects of Chinese herbal medicine Tianqi Pingchan Granule on G protein-coupled receptor kinase 6 involved in the prevention of levodopa-induced dyskinesia in rats with Parkinson disease.
Na WU ; Lu SONG ; Xinxin YANG ; Jianglei WEI ; Zhenguo LIU
Journal of Integrative Medicine 2012;10(9):1018-24
To investigate the effects of Tianqi Pingchan (TQPC) Granule, a compound traditional Chinese herbal medicine with antitremor activity, on levodopa-induced dyskinesia and the expression of G protein-coupled receptor kinase 6 (GRK6) in rats with Parkinson disease (PD).
5.Biocompatibility of poly-L-lactic acid/hydroxyapatite biomaterial with periodontal ligament cells
Ning ZHAO ; Xianjun ZHANG ; Wei LI ; Xinxin DONG
Chinese Journal of Tissue Engineering Research 2016;20(12):1732-1737
BACKGROUND:Hydroxyapatite/poly-L-lactic acid composite can wel improve the brittleness and mechanical properties of the hydroxyapatite. OBJECTIVE:To study the biocompatibility of hydroxyapatite/poly-L-lactic acid composite biomaterials with periodontal ligament cels. METHODS:Passage 4 human periodontal ligament cels at a density of 5×109/L were cultured with hydroxyapatite/poly-L-lactic acid composite as experimental group, and human periodontal ligament cels cultured alone as control group. Cel proliferation was detected within 7 days of culture; alkaline phosphatase activity was detected at 24, 48 and 72 hours of culture; and type I colagen expression was detected at 72 hours of culture. RESULTS AND CONCLUSION:There was no significant difference in the absorbance value of cels between the two groups at 1-7 days of culture. The alkaline phosphatase activity of cels at 48 and 72 hours was significantly higher than that at 24 hours (P < 0.05) in the two groups, but there was no difference between the two groups. The expression of type I colagen had no difference between the two groups. These findings indicate that the hydroxyapatite/poly-L-lactic acid composite biomaterial has good cel compatibility.
6.The inhibitory role of wogonin on tumor growth and telomerase activity of human ovarian cancer SKOV3 cell line xenograf in nude mice
Danrong LI ; Hanying ZHANG ; Wei ZHANG ; Li LI ; Xinxin HUANG
Chinese Pharmacological Bulletin 1986;0(04):-
Aim Telomerase is highly expression in most tumor cells, and it is an ideal target for cancer molecular targeting therapy. It has been proved that wogonin effectively inhibits telomerase activity and tumor cell growth in vitro. The study was to explore the inhibitory effect of wogonin on the growth of tumor and telomerase activity of implanted human ovarian cancer cell line SKOV3 in nude mice. Methods Nude mice with implanted human ovarian cancer cells SKOV3 were randomly divided into five groups, viz. the high dose group of Wogonin(600 mg?kg-1),low dose group of Wogonin(300 mg?kg-1),normal control group, cisplatin therapy group(3 mg?kg-1), and combined therapy group(cisplatin plus wogonin).The weight of nude mice and the volume of tumor were regularly measured. DNA、RNA and protein were extracted from the tumor tissue. The length of telomere was examined by Southern blot. The expression of telomerase hTERT gene was detected by RT-PCR. The telomerase activity was examined by TRAP-PCR-silver staining. Results The wogonin significantly inhibit the growth of tumor when compared with controlled group.The inhibitory rate of high dose group and low dose group were 56.67% (P=0.002) and 38.10%(P=0.019), respectively. The inhibition rate of cisplatin therapy group was 50.83%(P=0.004). The suppress rate of combined group reached 66.9% and higher than any single therapy(P=0.002). The length of telomere in different concentration groups of wogonin was the same as that in the control group.Wogonin inhibited the expression of telomerase gene hTERT and telomerase activity. The inhibition is related to the dose of wogonin. Conclusion Wogonin suppresses the growth and telomerase activity of tumor. The inhibitory effect is related to the dose of wogonin. Combination of wogonin and cisplatin increase the inhibitory rate in nude mice tumor.
7.Effects of propofol treatment on neuroapoptosis in primary cultured cortical neurons
Jianli LI ; Wei LIANG ; Xinxin PANG ; Honghai WU ; Yanning HOU
The Journal of Clinical Anesthesiology 2016;32(5):491-494
Objective To investigate the effect of propofol exposure on neuroapoptosis in pri-mary cultured cortical neurons and its mechanisms.Methods Cortical neurons were primarily cultured for seven days,then divided into two groups:control group (treated with equal volume of 20% in-tralipid),propofol-treated group (treated with 500 μmol/L propofol).The neurons were treated for 12 h.The neuron viability was determined by MTT.Neuroapoptosis was identified by Hoechest 33 258 dying.Mitochondrial membrane potential was measured by the fluorescent dye rhodamine 123 (Rh123).Western blot was performed to detect the level of cyt-c and cleaved-caspase-3.Results Neu-rons survival rate (54.4%±6.4%)in the propofol group was significantly lower than that of control group (99.8% ± 4.1%) (P < 0.05 ), the rate of neuronal apoptosis (46.5% ± 5.3%) was significantly higher than that of control group (7.2%±0.9%)(P <0.05),mitochondrial membrane potential (59.6%±4.3%)was significantly lower than that of the control group (99.9% ± 5.7%) (P <0.05 ),cyt-C protein level (0.38 ± 0.03 )was significantly higher than that of control group (0.1 5±0.02)(P < 0.05 ),level of cleaved-caspase-3 protein level (0.46 ± 0.04)was significantly higher than that of control group (0.13±0.02)(P <0.05).Conclusion Propofol induces neuroapo-tosis in primary cultured cortical neurons,which is associated with the decreased level of MPP and the increase levels of cyt-c and cleaved-caspase-3.
8.Study of Dimethyl Carbonate Adsorption and Activation on Solid Bases by Fourier Transform Infrared Spectroscopy
Gongde WU ; Xiaoli WANG ; Qingnian DONG ; Wei WEI ; Yuhan SUN ; Shan NI ; Xinxin ZHOU
Chinese Journal of Analytical Chemistry 2010;38(3):405-408
The adsorption and activation of dimethyl carbonate on the surface of solid base were investigated by in situ FTIR, and the solid bases included magnesia, magnesium fluoride, Mg-Al mixed oxide and fluorine-modified Mg-Al mixed oxide. The FTIR results showed that dimethyl carbonate adsorbed on the surface of solid based by two modes of bidentate and unidentate complex. The bidentate was more active than the unidentate. Methoxyl group was formed from the adsorbed dimethyl carbonate on the surface of magnesia and Mg-Al mixed oxide. And fluomethyl group was formed from the adsorbed dimethyl carbonate on the surface of sodium fluoride. However, dimethyl carbonate on the surface of fluorine-modified Mg-Al mixed oxide showed preference for generating fluomethyl group. With the increasing of the treating temperature of samples, the methoxyl group was gradually formed on the surface. Accordingly, the fluorine-modified Mg-Al mixed oxide was found to be an excellent catalyst for methylation.
9.Simultaneous Determination of Silybin A and Silybin B in Rat Plasma and Pharmacokinetic Study
Yang CHU ; Wei LI ; Zhiwen LI ; Xinxin LI ; Xiaohui MA ; Shuiping ZHOU ; Yonghong ZHU
Chinese Herbal Medicines 2011;(4):304-309
Objective To investigate the bioavailability and pharmacokinetics of silybin A and silybin B in rats,respectively.Methods Following iv and ig administration of silybin to 20 Wistar rats,the plasma samples were collected at different time points up to 12 h.Sample pretreatment was involved in one-step protein precipitation with acetonitrile.Silybin A and silybin B were simultaneously determined by LC-MS/MS.Results After ig dosing silybin 28,56,and 112 mg/kg to rats,the t1/2β values were 5.48,5.08,and 5.73 h for silybin A,and 4.56,4.12,and 5.53 h for silybin B; The Cmax were 674.3,1349.4,and 2042.5 ng/mL for silybin A,and 671.0,1365.4,and 2066.2 ng/mL for silybin B; The Tmax were 0.20,0.23,and 0.20 h for silybin A,and 0.20,0.23,and 0.20 h for silybin B; The AUC were 454.4,845.9,and 1219.5 h·ng/mL for silybin A,and 432.0,817.1,and 1153.6 h·ng/mL for silybin B.The absolute bioavailabilities of silybin A and silybin B were 2.86% and 1.93%,respectively.Conclusion Silybin A and silybin B have very low bioavailability after ig administration,and there is no significant difference in the pharmacokinetic parameters between silybin A and silybin B,which indicates that the two diastereoisomers have similar pharmacokinetic behavior in rats.
10.Physical-chemical properties and enhancement effect of Integrin αvβ3/CCR2 dual targeted ultrasound microbubbles:a vitro study
Yingying LIU ; Jinfeng XU ; Fei YAN ; Hui LUO ; Xinxin ZENG ; Zhanghong WEI ; Huiyu LIU
Chinese Journal of Ultrasonography 2015;(5):440-444
Objective To prepare Integrin αvβ3/CCR2 dual targeted microbubble,test physical-chemical properties,enhancement effect and targeting ability in vitro.Methods The dual targeted microbubbles (MBdual-target )with FITC labled iRGD and PE labled CCR2 were prapared,and non-target microbubbles as control (MBcontrol ) were prapered.Physical and chemical properties of two groups of microbubbles were tested,connectivity of peptides/antibodies and microbubbles were detected by fluorescence microscope and flow cytometry instrument.Enhancement effect and the stability of two groups of microbubbles was observed and compared in vitro.The affinity of MBdual-target and MBcontrol for bEnd.3 cells was investigated with light and fluorescent microscope.Results ①The particle size of MBdual-target was (0.93±0.23)μm,with no statistically significant difference compared with MBcontrol (P >0.05).②MBcontrol showed no fluorescent,while MBdual-target showed both clear green and red light,under fluorescent microscope.③There was no significant difference of gray scale of enhancement between MBdual-target and MBcontrol in vitro.④ It was showed that MBdual-target adhered to bEnd.3 cells in vitro experiment.Conclusions Integrinαvβ3/CCR2 dual targeted targeted microbubbles was successfully prepared and proved having good enhancement effect and targeting ability in vitro.