1.Change of endogenous hydrogen sulfide pathway in the monocrotaline-induced pulmonary hypertensive rats
Shasha FENG ; Wen YU ; Shuxu DU ; Junbao DU ; Hongfang JIN
Chinese Journal of Applied Clinical Pediatrics 2016;31(19):1489-1492
Objective To investigate the production of hydrogen sulfide (H2S) in rats with monocrotaline (MCT)-induced pulmonary hypertension(PAH).Methods Fourteen male Wistar rats were randomly divided into 2 groups (7 cases for each group):control group and MCT group.Rats in the MCT group were intraperitoneally injected with MCT (60 mg/kg) on day 1 while rats in the control group received only the same volume of 9 g/L saline.And then the conventional breeding was given for 21 days.Mean pulmonary artery pressure (mPAP) was evaluated via right cardiac catheterization procedure.The ratio of right ventricular mass (RV) and left ventricular plus septal mass (LV + SP) [RV/(LV + SP)] was calculated.The morphological change of pulmonary artery was observed by applying hematoxylin-eosin (HE) staining on the lung tissue paraffin section by measuring relative media thickness (RMT) and relative media area (RMA) of pulmonary aetery.H2S contents in serum and lung tissue rat were detected by using free radicals detector.The expression of cystathionine-γ-lyase (CSE),a key enzyme catalyzing endogenous H2S generaion,in lung tissue was detected by using Western blot method.CSE mRNA level in lung was detected by adopting the real-time polymerase chain reaction (RT-PCR) method.Results Compared with control group,mPAP of rats in MCT group was increased significantly [(49.31 ±3.67) mmHg vs (14.31 ±2.07) mmHg(1 mm Hg =0.133 kPa),P < 0.01] and the ratio of RV/(LV + SP) was increased (0.43 ± 0.03 vs 0.21 ± 0.03,P < 0.01),while RMT and RMA of pulmonary artery were enlarged [(43.46 ± 1.94) μm vs (13.16 ± 1.48) μm,P < 0.01;(3 321.10 ± 318.20) μm2 vs (963.40 ± 127.26) μm2,P <0.01].The evaluation of microstructure of pulmonary artery showed that the vessel wall of pulmonary artery in control rats was thin and no inflammatory cell was observed around artery.However,the vessel wall of pulmonary artery in rats of MCT group was thickened and there was evident infiltration of inflammatory cells in perivascular area.Compared with control group,H2S contents in serum and lung tissue of rats in MCT group were markedly decreased [(9.28 ± 0.94) μmol/L vs (14.20 ± 1.21) μmol/L,P < 0.01;(0.43 ± 0.08) μmol/g protein vs (0.87 ±0.17) μmol/g protein,P <0.01],while CSE protein and mRNA expression in lung tissue of rats in MCT group were downregulated (0.14 ± 0.02 vs 0.28 ± 0.09,P < 0.01;0.84 ± 0.06 vs 1.12 ± 0.04,P < 0.01).Condusion Endogenous H2S pathway is significantly downregulated in rats with monocrotaline-induced pulmonary hypertension.
2.Transplantation of bone marrow mesenchymal stem cells affects the proliferation and function of CD4+T cells in mice
Shaohong SU ; Junfeng ZHANG ; Qianru LI ; Shasha GUAN ; Ying DU
Chinese Journal of Tissue Engineering Research 2014;(1):106-111
BACKGROUND:Mesenchymal stem cells are found to have the immunoregulatory activities and a potential application prospect in the treatment of autoimmune diseases.
OBJECTIVE:To explore the mechanism of transplanting mesenchymal stems cells on the treatment of multiple sclerosis.
METHODS:The mouse mesenchymal stems cells were prepared, and injected into the al ogenic and syngenic normal mice, to detect the frequency of CD4+CD25+Foxp3+T cells in the spleen, thymus, and lymph nodes by flow cytometry, and to detect the Foxp3, transforming growth factor-β1, and interleukin-10 mRNA in the spleen, thymus, and lymph nodes by reverse transcription-PCR.
RESULTS AND CONCLUSION:Transplantation of mesenchymal stem cells on normal mice led to a significant up-regulation of CD4+CD25+Foxp3+T cells, Foxp3, transforming growth factor-β1, and interleukin-10 mRNA in the spleen, thymus, and lymph nodes both in the al ogenic and syngenic transplant groups. Transplantation of mesenchymal stem cells may be an available method in the treatment of autoimmune diseases, and CD4+CD25+Foxp3+T cell, Foxp3, transforming growth factor-β1, and interleukin-10 may be involved in this process.
3.Expression of Ten ABC Transporters in Cisplatin-resistant Nasopharyngeal Carcinoma Cell Lines
Siming XIE ; Shasha DU ; Weiyi FANG ; Kaitai YAO
Progress in Biochemistry and Biophysics 2006;0(04):-
Ten ATP binding cassette (ABC) transporters are confirmed to be associated with resistance against anticancer drugs. To investigate the relationship between these ten ABC transporters and the nasopharyngeal carcinoma cell line CNE2 resistant to cisplatin, cisplatin and cisplatin with 5-fluorouracil were used to induce the CNE2 cell to acquire the drug-resistance for 1 year. After these cells were cultured without drugs for 2 months, the MTT assay method was used to determine the dose-effect relationship of cisplatin and resistant index. Quantitative real time polymerase chain reaction was used to detect the mRNA expression of ten ABC transporters in CNE2 and the drug-resistant CNE2 cells, and the result was confirmed by immunocytochemical method. The results of MTT method showed that two cell lines resistant to cisplatin (named as CNE2/DDP) and cisplatin with 5-fluorouracil (named as CNE2/DDP+5Fu) were established, with resistant index 2.58 and 5.31, respectively. Of ten ABC transporters, only ABCC2 was found to be up-regulated both in CNE2/DDP and CNE2/DDP+5Fu cells, for increasing about 2.50 and 4.08 folds, respectively. The results of immunocytochemical method also confirmedthat the expression of ABCC2 in CNE2/DDP and CNE2/DDP+5Fu cells were stronger that that in CNE2 cell. Furthermore, ABCC2 protein was found to be located at nuclear membrane of CNE2/DDP +5Fu cell but not at nuclear membrane of CNE2 cell. The results suggest that ABCC2 may play an important role in cisplatinresistance of nasopharyngeal carcinoma cell line CNE2.
4.Influence of narrative therapy on the family environment and values in adolescents with emotional disorder
Zhao ZHAO ; Wendong DU ; Yixin CHEN ; Tunong CHEN ; Dandan WU ; Shasha LI ; Simeng GU
Chinese Journal of Behavioral Medicine and Brain Science 2012;(12):1082-1084
Objective To explore the effect of narrative therapy on family environment and values in adolescents with emotional disorder.Methods 35 adolescents with emotional disorder and their parents from Affiliated Brain Hospital,Nanjing Medical University were enrolled and treated by narrative therapy.Family environment was assessed using Family Environment Scale-Chinese Version (FES-CV)and was compared between pre-and posttherapy.Results 28 adolescents with emotional disorder and their parents completed therapy.The cohesion,independence,intellectual orientation,active-recreation orientation and organization in FES-CV showed higher scores at post-therapy than at pre-therapy ((6.89 ± 2.56) vs.(4.21 ± 3.21) ; (5.54 ± 1.35) vs.(4.68 ± 1.51) ;(4.75±2.07)vs.(3.50±2.12);(4.07±2.65)vs.(2.50±2.32);(3.68±1.77)vs.(2.50±1.62)respectively).The contradict in FES-CV showed lower scores at post-therapy than at pre-therapy ((2.75 ± 2.44) vs (5.25 ± 2.50)).The values of adolescents with emotional disorder and their parents had a positive change after narrative therapy.Conclusion Narrative therapy is effective on the positive change of FES-CV and values of adolescents with emotional disorder and their parents.
5.Mir-483 enhances the proliferation,invasion and migration of nasopharyngeal carcinoma cells
Miaohong YAN ; Shasha DU ; Rong LI ; Jie LIN ; Tan CHEN ; Yawei YUAN
The Journal of Practical Medicine 2017;33(2):177-180
Objective To investigate the effects of mir?483 on the proliferation,invasion and migration of nasopharyngeal carcinoma cells . Methods RT?qPCR was used to detect mir?483 expression level in different nasopharyngeal carcinoma cells(CNE?1,CNE?2)and the alteration of mir?483 expression level in these cells after radiation. CNE?1,one of the NPC cells,was selected. Cationic liposomes transient transfection was used to construct cells which overexpressed mir?483 or negative control . MTT assay was conducted to test the cell proliferation ability. Wound scratch assay and transwell migration assay were used to detect the effects of mir?483 on NPC invasion and migration ability. Results In terms of radiation resistance ,the expression level of mir?483 was higher in CNE?1. Overexpressing mir?483 can enhance the proliferation ,invasion and migration ability of CNE?1. Conclusions Mir?483 enhances the proliferation,invasion and migration ability of nasopharyngeal carcinoma cells.
6.Protective effect of tanshinoneⅡA on radiation injury of cochlea
Peixin TAN ; Shasha DU ; Chen REN
The Journal of Practical Medicine 2017;33(17):2821-2826
Objective To explore the protective effects on radiation injury of guinea pigs 'cochlea by sul-fotanshinoneⅡA sodium injection. Methods 20 guinea pigs(40 ears)were randomly divided into control group , radiotherapy(RT)group,radiotherapy and drug(RT+D)group and drug(D)group. The guinea pigs in the RT+D group and the D group were injected intraperitoneally with tanshinone on 2 days before the irradiation and last-ed for 7 days,while the guinea pigs in the control group and the RT group were injected intrapertoneally with equal amount of physiological saline at the same time. All the guinea pigs underwent audiologic test with DPOAE and ABR at different time points before and after the irradiation,which were d0,d7 and d8w. All pigs were sacrificed after the completion of the audiologic experimental and for histologic observation. Results After the irradiation, the DPOAE amplitudes of the guinea pigs in both RT group(F=1134.064,P<0.001)and RT+D group(F=664.185,P < 0.001)decreased significantly,but the range in the RT + D group was lower than in the RT group (P<0.001). The ABR threshold of the guinea pigs raised significantly in the RT group(F=12.228,P=0.002 <0.05),but not in the RT+D group(F=2.867,P=0.102>0.05). Histological examination showed that the dam-age degree of hair cells,stria vascularis and the spiral ganglion cells in the organ of Corti of the guinea pigs were lower than in the RT + D group,but higher in the RT group. Conclusion Sulfotanshinone ⅡA sodium injection can provide protection against radiation injury of guinea cochlea.
7.Service quality of primary healthcare institutions under the vertically integrated model: A perspective of demanding side
Shasha YUAN ; Zhipeng YONG ; Fang WANG ; Jun ZHAO ; Xi LI ; Yongchao CHEN ; Hongyi DU
Chinese Journal of Health Policy 2017;10(7):41-46
Objective: On the basis of typical cases, the objective of this study is to assess the service quality of primary healthcare institutions under the vertically integrated model from the perspective of demanding side.Methods: The vertical model between Renhe Hospital and Lixian Township Health Center was selected as the case study.Quantitative data were collected by exit review using Primary health-Care quality service Assessment Tool (PCAT)-Chinese version from the patients seeking services in Lixian.Quantitative data were analyzed by descriptive statistics and multiple linear regression equations while qualitative data were analyzed by thematic framework analysis.Results: Under the vertical model, the PCAT total score was 20.09.The scores on the first diagnosis, continuity of care, and comprehensiveness of care and coordination of care were 6.59, 3.27, 4.58 and 5.62 respectively.The scores on the three dimensional extensions, family-centered, community-oriented, medical culture were 3.07, 1.86 and 2.67 respectively.The difference was significant for the PCAT total score between a number of visits and the length of waiting time for services in Lixian.Conclusions: Under the vertical model, primary healthcare institutions did better in the first diagnosis while weaker in continuity and the three dimensional extensions.With the development of the integration, the primary healthcare institutions need to strengthen the continuity and downward referral in the future.Future researches and follow-up, analysis is needed to track the impact of the vertically integrated model.
8.Audiological Characteristics in 832 Deaf Children with Biallelic Causative Mutations in GJB2,SLC26A4 Gene
Qingjia CUI ; Guojian WANG ; Yuan ZHANG ; Ying YANG ; Dongyang KANG ; Yanshun DU ; Liping ZHAO ; Shasha HUANG ; Wei ZHANG ; Xibin SUN ; Pu DAI ; Lihui HUANG
Journal of Audiology and Speech Pathology 2014;(2):120-123
Objective To determine the audiological characteristics in 832 deaf children with biallelic causative mutations in GJB2 ,SLC26A4 gene .Methods The 832 patients received deafness gene screening ,553 were GJB2 gene biallelic causative mutations ,279 were SLC26A4 gene biallelic causative mutations .Patients were divided into four groups according to ages of hearing loss onset :<1 ,1~3 ,3~6 ,6~12 years old ,and the audiological character-istics and prevalence of GJB2 ,SLC26A4 gene mutations at different ages of onset .Results The prevalence of GJB2 gene mutations at four groups was 37 .97% (210/553) ,38 .34% (212/553) ,16 .27% (90/553) ,7 .41% (41/553) ,re-spectively ;the prevalence of SLC26A4 gene mutations at four groups was 25 .45% (71/279) ,44 .80% (125/279) , 20 .07% (56/279) ,9 .67% (27/279) ,respectively .The difference between GJB2 and SLC26A4 gene was significant(P=0 .001) .The prevalence of profound hearing loss with GJB2 gene mutations at four groups were 66 .67% (140/210) ,61 .32% (130/212) ,47 .78% (43/90) ,41 .46% (17/41) ,respectively .The difference was significant (P=0 .004) ,while the difference in 279 patients with SLC26A4 gene mutations was not statistically significant (P= 0 . 083) .Conclusion The age of hearing loss onset in patients with biallelic causative mutations in GJB 2 or SLC26A4 gene refers to 0~3 years -old ,hearing loss in patients with GJB2 ,SLC26A4 gene mutations gives priority to pro-found .The age of hearing loss onset is smaller ,the ratio of profound hearing loss is higher .Patients with severe and profound hearing impairment should be performed the genetic testing when the age of onset under 12 .
9.The effect of uric acid on the proliferation and phenotypic change of rat glomerular mesangial cells in vitro
Shasha LI ; Xiaoli GUO ; Yan DU
The Journal of Practical Medicine 2019;35(3):392-397
Objective To investigate the effect of different concentration of uric acid on proliferation and phentyic change of HBZY-1 cells in vitro. Methods The morphological change of HBZY-1 cells exposed to uric acid was observed under the inverted microscope and transmission electronmicroscope. MTT bioassay was used to assess the effect of uric acid on the proliferation of HBZY-1 cell. ELISA method was used to assess the level of angiotensin Ⅱ in HBZY-1 cell incubated with different concentration of uric acid for different treatment duration.The protein and mRNA expression of α-smooth muscle actin (α-SMA) , transforming growth factor-β1 (TGF-β1) and fibronectin in HBZY-1 cells was measured by Western blot and RT-PCR assay, respectively. Results Uric acid (0.4 mmol/L, 24 h) could induce phentypic change of messangial cell from triangle to fibroblast-like morphology.Uric acid could significantly stimulate the proliferation of HBZY-1 cells for 24 h in a concentration-dependent manner (P < 0.05). Uric acid could stimulate the secration of angiotensin Ⅱ in a dose-and time-dependent manner.RT-PCR and Western blot results showed that the markers of myofibroblast transdifferentiation, including a-smooth muscle actin (α-SMA) , transforming growth factor-β1 (TGF-β1) and fibronectin, were significantly upregulated in HBZY-1 cell treated with over 0.1 mmol/L uric acid for 48 h compared with the control group (P < 0.05, respectively). Conclusion Uric acid can induce the proliferation and phentypic change of HBZY-1 in vitro.
10.Separation and identification of chemical constituents of Mongolian medi-cine compound Shudage-4
Shasha XIN ; Yinfei DU ; Xin JIA ; Yang YANG ; Yu DONG
Journal of Beijing University of Traditional Chinese Medicine 2016;39(11):941-944
Objective To study the chemical constituents of Mongolian medicine compound Shudage-4, so as to find out material bases of its pharmacodynamic effects.Methods The chemical constitutes of Shudage-4 were extracted using ethanol extraction method,and were isolated and purified using technolo-gies of silica gel,Sephadex LH-20 and reversed phase silica gel.Then the structures of the isolated com-pounds were identified by using different spectroscopic and chromatographic techniques including ultra vi-olet spectroscopy(UV),infrared spectroscopy(IR),mass spectrometry(MS)and nuclear magnetic reso-nance(NMR).Results There were 10 compounds isolated and identified from Mongolian medicine Shudage-4—1-phenyl-7-(3-methoxy-4-hydroxy) phenyl-5-ol-3-heptanone-one (1 ),stigmasterol (2 ), kaempferol(3),daucosterol(4),p-Anisic acid(5),ursolic acid(6),oleanic acid (7),dehydrocostus lactone(8),costunolide(9)and dehydrodiisoeugenol (10).Conclusion The ten compounds,except stigmasterol,were first isolated from Shudage-4.This paper provided important scientific data for the ma-terial bases of Mongolian medicine Shudage-4.