1.Role of autophagy in human mesenchymal stem cells in response to irradiation
Zhe CHEN ; Jie LIU ; Bin ZHANG
Chinese Journal of Tissue Engineering Research 2014;(28):4474-4478
BACKGROUND:Bone marrow mesenchymal stem cells can survive under the lethal dose of radiation in response to hematopoietic stem cells, and stil maintain the typical characteristics of stem cells to promote hematopoietic recovery after radiation. However, autophagy is an important mechanism for cellular adaptation under stress, which may be involved in radiation tolerance of bone marrow mesenchymal stem cells. OBJECTIVE:To investigate the role of autophagy in human bone marrow mesenchymal stem cells in response to irradiation. METHODS:Passage 3 bone marrow mesenchymal stem cells cultured in vitro at logarithmic phase were col ected and randomized into control, 3-mehyladenine, rapamycin, irradiation, irradiation+3-mehyladenine and irradiation+rapamycin groups. The autophagy reactions of bone marrow mesenchymal stem cells were regulated by 5 mmol/L 3-mehyladenine and 200 nmol/L rapamycin for 12 hours in the 3-mehyladenine and rapamycin groups, respectively. Two-hour 6 Gy X-ray irradiation was performed in the irradiation group and two complication groups undergoing 12-hour corresponding drug intervention. RESULTS AND CONCLUSION:The proportions of cells with autophagic vacuoles and apoptotic cells were higher in the irradiation group than the control group, moreover, autophagic+apoptotic cells were increased in the irradiation group. 3-mehyladenine intervention could decrease the proportion of cells with autophagic vacuoles,and increase the number of apoptotic cells. But there was no difference in the proportion of autophagic+apoptotic cells between the 3-mehyladenine and irradiation groups. After rapamycin intervention, the proportion of autophagic cells was higher than that in the irradiation group, but no difference in the proportion of apoptotic cells between the two grups, as wel as there were no autophagic+apoptotic cells. The expression of microtubule-associated protein 1 light chain 3-II was ranked as fol ows:the control and 3-mehyladenine groups
2.Establishment and characterization of a multidrug resistant human mucoepidermoid carcinoma xenograft model
Xiaofang XU ; Bin LIU ; Junzheng WU ; Chunmei WANG ; Zhe WANG
Journal of Practical Stomatology 2010;26(2):177-180
Objective:To establish a multidrug resistant model of human mucoepidermoid carcinoma using nude mice. Methods:Multidrug resistant MEC/5-FU cells were inoculated intradermally into nude mice. Solid tumors were locally measurable after 10 days and 5-FU was repeated intraperitoneal injected into tumor-bearing mice. The tumor cells in nude mice (MEC/5-FU/NU) were isolated, cultured and examined. Results:The xenografts were similar to the original mucoepidermoid carcinoma from which the cell line was derived. The resistance index (RI) of the MEC/5-FU/NU cells to 5-FU was 27.82. Compared to the MEC, the expressions of ABCB1, ABCB11 and GSTA1 genes and MDR-1 protein increased in the MEC/5-FU/NU cells(P<0.05). Conclusion:The xenograft model is a good model of human multidrug resistant mucoepidermoid carcinoma, and may be useful in studying drug resistance mechanism in vivo.
3.Designation and evaluation of antisense oligodeoxynucleotides targeted to glial glutamate transporter-1a.
Li-zhe LIU ; Min ZHANG ; Yi-xian LIU ; Xin CUI ; Yu-yan HU ; Wen-bin LI
Chinese Journal of Applied Physiology 2015;31(3):238-243
OBJECTIVEThe present study was undertaken to design antisense oligodeoxynucleotides (AS-ODNs) of glial glutamate transporter-la (GLT-1a) and to evaluate the effectiveness of the designed AS-ODNs on the expression of GLT-1a.
METHODSFive sequences of GLT-1a AS-ODNs were designed according to the C terminus specific sequences of GLT-1a mRNA using antisense design software of IDT Com- pany. Western blot analysis was used to evaluate the inhibition effects of the five GLT-1a AS-ODNs on the expression of GLT-la.
RESULTSThe sequence of GLT-1a AS-ODNs with sequence of 5'-GGTTCTTCCTCAACACTGCA-3' could specifically inhibit the expression of GLT-1a in the hippocampal CA1 subfield of rats, while it had no effect on the expression of GLT-1b. This sequence showed similar inhibition on the expression of GLT-la in sham and ceftriaxone (Cef)-treated rats. It could also significantly inhibit the cerebral ischemic preconditioning (CIP)-induced up-regulation in the expression of GLT-1a. The magnitude of the inhibition in sham, Cef- or CIP-treated rats was similar by more than 60%.
CONCLUSIONFrom the designed five sequences of GLT-1a AS-ODNs, we obtained an effective sequence which can specifically inhibit the expression of GLT-1a.
Animals ; CA1 Region, Hippocampal ; metabolism ; Excitatory Amino Acid Transporter 2 ; antagonists & inhibitors ; metabolism ; Ischemic Preconditioning ; Oligonucleotides, Antisense ; genetics ; RNA, Messenger ; Rats ; Up-Regulation
5.The clinical significance of adjuvant radiotherapy in Mucin1 mRNA-positive patients following Ivor-Lewis esophagectomy in esophageal cancer
Bin SHANG ; Zhe YANG ; Huaxia CHEN ; Xiangyan LIU ; Gang CHEN ; Zhou WANG
Chinese Journal of Thoracic and Cardiovascular Surgery 2015;31(2):88-92
Objective To investigate whether Ivor-Lewis esophagectomy combined with adjuvant radiotherapy prevents lymphatic metastatic recurrence in esophageal cancer patients.Methods 113 Stage Ⅱ A esophageal squamous cell carcinoma patients after Ivor-Lewis esophagectomy were accpected mRNA expression of Mucin1 gene detection.Positive patients were enrolled into adjuvant radiotherapy group(with postoperative adjuvant radiotherapy).Negative patients were enrolled into control group (without postoperative adjuvant radiotherapy or chemotherapy).The radiotherapy area consisted of the neck,supraclavicular region and the superior mediastinum(including praesophageal and pratracheal region).Survival difference was compared by x2 test,the Kaplan-Meier method was performed to calculate the survival rate and recurrence rate.Logistic regressive analysis was performed to determined independent risk factors.Results The radiotherapy area lymphatic metastatic recurrence rate in adjuvant radiotherapy group(16.7%,5/30) was lower than patients without postoperative adjuvant radiotherapy (45.8%,38/83) (P < 0.05).Only compared to positive patients without postoperative adjuvant radiotherapy(60.0%,6/10),the rate (16.7%,5/30) was significantly lower(P < 0.01).Cancer recurrence was recognized in 48.6% (55/113) patients within 3 year after operation,including 38.1% (43/113) patients with radiotherapy area recurrence.In logistic analysis the T status (P< 0.01) and adjuvant radiotherapy (P < 0.05) were independent risk factors of lymph node metastasis in the first 3 years after operation.Conclusion In Mucin1 mRNA-positive esophageal squamous cell carcinoma patients,adjuvant radiotherapy could significantly reduce the lymph node metastasis rate in the radiotherapy area after Ivor-Lewis esophagectomy.Compared with traditional therapeutic methods,Ivor-Lewis esophagectomy combined with adjuvant radiotherapy can achieve similar curative effects in Mucin1 mRNA-positive patients.
6.Antioxidant response element activator protects motor neurons from selected death
Hui BU ; Chun-Yan LI ; Xiao-Yun LIU ; Zhe LI ; Bin LI ; Meng-Meng SUN
Chinese Journal of Neurology 2001;0(01):-
Objective To investigate the effects of antioxidant response element (ARE) activator- 5,6-dihydrocyclopenta[ C ]-1,2-dithiole-3-thione (CPDT) on organotypic spinal cord cultures and to study whether this activation can protect motor neurons from oxidative stress.Methods Organotypic spinal cord cultures were prepared using lumbar spinal cord slices from 8-day-old rat.Threo-hydroxyaspartate (THA) was continuously added into the culture medium for 3 weeks,which caused selective motor neuron death. Thus,the in vitro model of amyotrophic Lateral sclerosis (ALS) was established.15,30 ?mol/L of CPDT were added into the culture medium respectively.Ventral motor neurons survival was evaluated by immunohistochemical staining with monoclonal antibody SMI-32,a nonphosphorylated neurofilament marker. Ultrastructure was observed with electronic microscope.Results The pretreatment of organotypic spinal cord cultures with different concentrations of CPDT significantly increase the total number of ventral motor neurons (15?mol/L:(15.81?6.97) perexplant;30?mol/L:(16.25?6.74) perexplant respectively) compared with THA group ((5.31?5.76) perexplant) and the former had plentiful neurite extensions (n= 15,P
7.Anterior internal lenticonus accompanied by congenital nuclear cataract.
Zhe LIU ; Chuan-Bin SUN ; Ke YAO
Chinese Medical Journal 2011;124(23):4119-4120
Internal lenticonus is a very rare morphologic abnormality of crystalline lens which has been reported in only several cases in the literature. We herein reported the clinical characteristics and surgical findings of the anterior internal lenticonus accompanied by congenital nuclear cataract. Cataract extraction accompanied with intraocular lens implantation was uneventfully performed, and a good visual outcome was achieved in this case. Viral infection during embryonal and fetal period might account for the formation of the anterior internal lenticonus and congenital nuclear cataract in our case.
Cataract Extraction
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methods
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Humans
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Lens Diseases
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surgery
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Lens Implantation, Intraocular
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methods
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Lens, Crystalline
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abnormalities
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surgery
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Male
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Middle Aged
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Visual Acuity
8.Dynamic changes of HSP70 mRNA expression in liver tissue of rats with traumatic shock and the treatment effects of glycine
Zhe DENG ; Dehong LIU ; Yugang XIE ; Zhongjiang ZHAO ; Yiwu SUN ; Zheqiang ZHOU ; Bin YAO ; Min QIU ; Weidong WANG
Journal of Chinese Physician 2008;10(6):739-742
Objective To investigate the dynamic changes of HSP70 mRNA expression in the liver tissue of rats with traumatic shock and the treatment effects of glycine.Methods The expression of HSP70 mRNA in the liver tissue of treatment group,shock group and control group was detected by ELISA.Pathological changes were observed,and serum ALT and AST were measured.Results The expression of HSP70 mRNA in the liver tissue of rats in the shock group and the treatment group reached peak at the 6th and 12th hour after resuscitation respectively.Serum ALT and AST increased and pathological damage aggravated with time prolonging.Compared with control group,the expression of HSPT0 mRNA in treatment group increased significantly,serum ALT and AST decreased significantly and pathologi- cal damage was significantly relieved(all P<0.05).Conclusion Glycine can increase the expression of HSPT0 mRNA and relieve the secondary damage of liver after traumatic shock.
9.Protective mechanisms of glycine against the secondary liver injury of rats after taumatic shock
Zhe DENG ; Xinjian YANG ; Zhongjiang ZHAO ; Zeqiang ZHOU ; Dehong LIU ; Yugang XIE ; Jiwu SUN ; Bin YAO ; Xiaoying ZHENG
Chinese Journal of Trauma 2009;25(8):739-742
Objective To investigate the effects of glycine on the expression of HSP70 and TNF-α mRNA in the liver tissue of rats with traumatic shock and explore the protective mechanism of glycine a-gainst secondary liver injury after traumatic shock. Methods The traumatic shock model was established and 120 Wistar rats were divided randomly into three groups: treatment group, shock group and control group. At the beginning of resuscitation, the rats in the treatment were injected with 0.5 ml isotonic saline containing 100 mg/kg glycine, those rats in the shock group were injected only with 0.5 ml isotonic saline. The rats in three groups were killed at 3, 6, 12, 24 and 48 hours after resuscitation respectively. The ex-pression of HSP70 and TNF-α mRNA in the liver tissue were detected by RT-PCR, pathological changes were observed and serum ALT and AST were measured. Results The expressions of HSP70 and TNF-α mRNA in the liver tissue of rats in the shock group began to increase at 3 hours and both reached the peak value at 6 hours after resuscitation, but the expression of HSP70 mRNA in the treatment group reached the peak value at 12 hours after resuscitation. Compared with the control group, the expression of HSP70 mR-NA in the treatment group increased significantly and that of TNF-α mRNA decreased siganicantly, serum ALT and AST decreased and pathological damage was relieved significantly (all P < 0.05). Conclusion By enhancing the expression of HSP70 mRNA and decreasing the expression of TNF-α mRNA, glycine may play a protective role against the secondary damage of liver after traumatic shock.
10.Effect of ethyl acetate extract of sappan wood on expression of myocardial GrB mRNA in rat model of allogeneic ectopic cardiac transplantation.
Jia-Xin ZHENG ; Ya-Bin ZHOU ; Ying-Zhe LIU
Chinese Journal of Integrated Traditional and Western Medicine 2008;28(6):537-540
OBJECTIVETo explore the action mechanism of the immune active components of sappon wood (SWE) for antagonizing reject reaction by observing the influence of its ethyl acetate extract on mRNA expression of myocardial GrB in rat model of allogenic ectopic cardiac transplantation.
METHODSAnimal model of abdominal cardiac ectopic transplantation was established taking Wistar rat as the donor and SD rat as the receptor. The successfully modeled rats were randomly divided into the model group, the SWE group and the CsA group. GrB mRNA expression was detected by RT-PCR method and myocardial pathomorphologic picture was observed in routine.
RESULTSThe pathologic changes in the SWE group (23 scores) and the CsA group (14 scores) were milder than in the model group (31 scores), the former two could markedly alleviate the myocardial pathologic injury (P<0.05, P<0.01). The GrB mRNA expression in the model group was 1.3000 +/- 0.1207, the SWE group 0. 7070 +/- 0.1215, and the CsA group 0.6700 +/- 0.0997, respectively; compared with the model group, the latter two could obviously down-regulate the expression of GrB mRNA (P<0.01) and no significant difference was found between the latter two groups (P>0.05).
CONCLUSIONSWE could alleviate the pathologic change, down-regulate the mRNA expression of myocardial GrB in allogenic ectopic transplanted myocardium of rats, it is possibly one of the factors for its antagonizing effect against reject reaction.
Acetates ; chemistry ; Animals ; Caesalpinia ; chemistry ; Gene Expression Regulation, Enzymologic ; drug effects ; Graft Rejection ; drug therapy ; Granzymes ; genetics ; Heart ; drug effects ; Heart Transplantation ; immunology ; Male ; Models, Animal ; Myocardium ; metabolism ; pathology ; Plant Extracts ; pharmacology ; therapeutic use ; RNA, Messenger ; genetics ; metabolism ; Rats ; Transplantation, Homologous