1.The effect of adenovirus-mediated ING4 expression on breast cancer cells
Zhengyi LI ; Weihua SHENG ; Yufeng XIE ; Huicui YANG ; Jicheng YANG
Chinese Journal of General Surgery 2010;25(4):313-317
Objective To investigate the inhibitory effect of inhibitor 4 of growth(ING4)delivered by adenovirus on human breast carcinoma cell MDA-MB-231.Methods MDA-MB-231 human breast carcinoma cells were irfected with Ad-ING4.The expression level of ING4 gene was detected by RT-PCR and Western blot;The growth inhibition,cell-cycle alteration,and apoptosis were detected by MTT,Flowcytometry and Hochest33258 staining.respectively.RT-PCR was used to detect the transcription of Bax,Bc1-2,Survivin genes;The expression level of Ang-1 gene was detected by ELISA;Ad-ING4 was given intratumorally in athymic nude mice beating MDA-MB.231 tumors.and then tumor growth was monitored;The expression of Bc1-2,Bax and Caspase-3 was analyzed by immumohistochemistry.Results ING4 was successfully transcribed and translated iU MDA-MB-231 cells:Ad-ING4 significantly inhibited the proliferation and induced G_2/M phase arrest to(24.86±1.24)% and cell apoptosis of MDAMB-231.Intratumoral injection of Ad-ING4 suppressed the tumor growth obviously with a inhibitory rate of 49%;Immumohistochemistry showed that the expression of Bax,Caspase-3 were up-regulated and the expression of Bc1-2.Survivin,CD34 were down-regulated by Ad-ING4.Conclusions Ad-ING4 can inhibit the growth of MDA-MB-231 cells and induce apoptosis in vitro and in vivo.
2.Preparation and Identification of Main Ingredients in Effective Parts of Xinjiang Artemisia rupestris
Xiaojuan RONG ; Zhengyi GU ; Jinhua HE ; Lu YANG
China Pharmacy 2017;28(16):2227-2230
OBJECTIVE:To establish a method for rapid identification and efficient preparation of main ingredients in effective parts of Xinjiang Artemisia rupestris,and provide reference for researching the ethnic medicines. METHODS:LC-HRMS/MS was conducted for the preliminary study of main ingredients in effective parts of A. rupestris. HPLC,UV and MS were used to compare and analyze parts of the compounds and its reference substances,their names were determined. Column separation and preparative liquid chromatography were used for the undetermined compounds to receive monomer rapidly,and the structures were identified. RESULTS:5 compounds were separated from the effective parts,2 of which were identified as artemetin and casticin. A monomer-ic compound was obtained (yield was 0.35 mg/g,the purity was 98.5%),which was confirmed to be 6-demethoxy-4′-O-me-thoxy-capillarisin-7-O-β-D-glucoside by the structure. CONCLUSIONS:The method has achieved rapid separation,identification and preparation of target ingredients,which can be used for the fundamental research of ethnic medicine complex materials.
3.Clinical manifestations and cerebral angiographic findings of moyamoya disease
Jianfeng HAN ; Chengtai WANG ; Jianbo YANG ; Feng GUO ; Zhengyi LI
Journal of Pharmaceutical Analysis 2010;22(2):131-134
Objective To study the clinical features and angiographic findings of moyamoya disease (MMD) as well as their relationship. Methods A total of 22 MMD patients received routine digital substraction angiography (DSA). The clinical manifestations and angiographic findings were analyzed. Results Clinical manifestations varied and each patient often had multiple symptoms, including cerebral infarction in 9 patients with an average age of 23.6 (13-39 years) and cerebral hemorrhage in 7 patients with an average age of 31.2 (28-46 years). Angiographic examination found that all the diseased sides showed MMD blood vessels. The patients who received encephalo-myo-arterio-synangiosis (EMAS) had better prognosis than those without receiving the treatment. Conclusion Cerebral infarction is frequent in children and adolescents with MMD, whereas cerebral hemorrhage is common in adults. DSA is a golden criterion for diagnosing MMD. Surgical treatment, EMAS blood supply reconstruction in particular, should be prescribed.
4.Left lower extremity venous thrombosis after operation of cyst in the jaw: a case report.
Yongchun YU ; Weiming YU ; Zhengyi YANG ; Shubiao ZHANG ; Jie SUN ; Lei LIU
West China Journal of Stomatology 2014;32(2):203-205
Deep vein thrombosis is a common complication after orthopedic, gynecological, and obstetric operations. However, instances of this complication after oral and maxillofacial surgeries have been rarely reported. We report a case with odontogenic keratocyst in the jaw that gained left lower extremity venous thrombosis after operation, and discuss the risk factors and prevention of lower extremity venous thrombosis through literature review.
Cysts
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surgery
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Humans
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Lower Extremity
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Mandibular Diseases
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surgery
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Risk Factors
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Venous Thrombosis
5.Correlations between stroke-preventing knowledge,health relief and health behaviors in community hypertensive patients
Lihong WAN ; Zhengyi YANG ; Lijuan LI ; Shiying CHENG ; Shuyi LIU ; Shan XIAO ; Wei TANG
Modern Clinical Nursing 2013;(12):1-5
Objectives To study the correlations between stroke-preventive knowledge,health belief and health behaviors in community hypertensive patients.Methods The questionnaire of SPKQ,CHBMS and HPLPⅡwere used to take the investigation among 94 hypertensive patients from a community hospital in Guangzhou.Results The total score on SPKQ was 62.70±18.39 and the average scores on CHBMS and HPLPⅡwere 3.51±0.24 and 2.48±0.37,respectively.The stroke-preventive knowledge was positively correlated with health belief,health motivation and self-efficacy(r=0.289,P<0.01;r=0.246,P<0.05;r=0.350 (P<0.01,respectively).The health motivation was positively correlated with health behaviors(r=0.304,P<0.01)and the seriousness negatively correlated with health behaviors(r=-0.279,P<0.01).Conclusion Medical staff should provide much more stroke education with community hypertensive patients and promote patients’health motivation and self-efficacy of health belief in stroke prevention,help patients understand stroke seriousness,establish and sustain healthy lifestyles.
6.Comparative study between mesenchymal stem cells derived form orofacial bone and those from long bone
Lin YUAN ; Han WANG ; Jin SUN ; Zhengyi YANG ; Lu HAO ; Guangsi PAN ; Yina CAO ; Jun QIAN
Journal of Practical Stomatology 2015;(4):526-530
Objective:To compare the proliferation and osteogenic differentiation between human bone marrow mesenchymal stem cells from orofacial bone(OMMSCs)and those from long bone(BMMSCs).Methods:OMMSCs were isolated from orthognathic surgical sites and cultured by limited dilution.BMMSCs were obtained from bone marrow of volunteers and isolated by density gradient centrifugation method.The surface markers of the cells were detected by flowcytometry.Single-colony formation,CCK assay and cell circle analyses were conducted.Osteogenic differentiation ability was evaluated by ALP activity test and Alizarin red staining after osteogenic induction culture.Results:The cell surface markers STRO-1 and CD105 of both stem cells were positive,CD34,CD31 and CD45 were negative. OMMSCs generated significantly higher numbers of colonies than BMMSCs.In addition,OMMSCs had a higher proliferation rate and more cells in proliferative(S +G2 )stage than BMMSCs.After osteogenic induction for 3,5,7 and 10 d,OMMSCs showed higher levels of ALP activity.OMMSCs formed significantly more mineralized nodules than BMMSCs after 21-day ostogenic induction.Conclusion:The proliferation and osteogenic differentiation capacity of OMMSCs are higher than those of BMMSCs.
7.Effect of morphine preconditioning on expression of miR-133b-5p and Fas in rat cardiomyocytes subjected to hypoxia/reoxygenation
Shufang HE ; Haijuan ZHU ; Jie CHENG ; Shijing XU ; Zhengyi HAN ; Wan YANG ; Ye ZHANG
Chinese Journal of Anesthesiology 2015;35(6):747-750
Objective To evaluate the effect of morphine preconditioning on the expression of miR-133b-Sp and Fas in rat cardiomyocytes subjected to hypoxia/reoxygenation (H/R).Methods Cardiomyocytes were isolated from healthy adult male Sprague-Dawley rats by using Langendorff perfusion.The cells were seeded into 24-well plates or 60 mm diameter dishes and randomly divided into 3 groups (n =24 each) using a random number table:control group (group C),group H/R,and morphine preconditioning group (group MPC).The cells in group C were cultured in normal culture atmosphere.In H/R and MPC groups,the cells were exposed to 95% N2-5% CO2 for 90 min followed by 120 min reoxygenation.In group MPC,the cells were cultured for 10 min in serum-free DMEM liquid culture medium containing morphine 1 μmol/L,and then were cultured for 30 min in morphine-free DMEM liquid culture medium before hypoxia.At 120 min of reoxygenation,the cells in 24-well plates were selected to detect the cell viability (by MTT),lactate dehydrogenase (LDH) activity in the culture medium,and cell apoptosis (by Hoechst 33234 staining).Apoptosis rate was calculated.Total RNA and protein were extracted from the cells in 60 mm dishes to detect the expression of miR-133b-5p and Fas mRNA (by quantitative real-time PCR) and Fas protein (by Western blot).Results Compared with C group,the cell viability was significantly decreased,LDH activity and apoptosis rate were increased,the expression of miR-133b-Sp was down-regulated,and the expression of Fas mRNA and protein was up-regulated in H/R group.Compared with H/R group,the cell viability was significantly increased,LDH activity and apoptosis rate were decreased,the expression of miR-133b-5p was up-regulatcd,and the expression of Fas mRNA and protein was down-regulated in MPC group.Conclusion The mechanism by which morphine preconditioning reduces H/R injury to rat cardiomyocytesis related to up-regulation of the expression of miR-133b-Sp and down-regulation of the expression of Fas.
8.Effects of morphine preconditioning on expression of microRNAs during hypoxia-reoxygenation in H9 c2 myocardial cells
Zhengyi HAN ; Shufang HE ; Jie CHENG ; Shijin XU ; Wan YANG ; Ye ZHAGN
Chinese Pharmacological Bulletin 2015;(11):1552-1557
Aim To evaluate the effects of morphine preconditioning ( MPC ) on the expression of microR-NAs ( miRNAs ) induced by hypoxia-reoxygenation (H/R) in H9c2 myocardial cells. Methods H9c2 cells were randomly divided into 3 groups ( n=4 each) as follows:control group ( CON) , hypoxia/ reoxygen-ation group ( H/R ) and morphine preconditioning group ( MPC+H/R) . The cells were cultured in nor-mal condition in CON group. The cells were subjected to 5 h hypoxia followed by 1 h reoxygenation in H/R group and MPC+H/R group. Specifically, the cells in MPC+H/R group were preconditioned with morphine with the final concentration of 1 μmol·L-1 for 10 min before H/R. After the treatment, CCK-8 was used to detect cell viability and chemical colorimetry was used to detect lactate dehydrogenase ( LDH ) activity in the culture medium. Cell apoptosis was assessed by An-nexin-V-FITC/PI flow cytometry. Relative expression of Fas protein was detected by Western blot. The ex-pression of miRNA in myocardial cells was analyzed by quantitative reverse transcription polymerase chain re-action ( qRT-PCR ) . Results Compared with CON group, the cell viability was significantly decreased, while the LDH activity, apoptotic rate and Fas protein expression were dramatically increased in group H/R (P<0. 01). However, MPC significantly increased the cell viability, whereas it decreased the LDH activity, apoptotic rate and Fas protein expression induced by H/R injury ( P < 0. 01 ) . The expressions of miR-133a-5p, miR-133b-5p, miR-664-1-5p, miR-6216 and let-7 e-5 p were markedly down-regulated by H/R as compared to CON group ( P <0. 05 ) , while MPC inhibited these miRNAs which were significantly down-regulated by H/R group ( P <0. 01 ) . Conclusion Morphine preconditioning might protect H9 c2 myocar-dial cells against H/R injury by regulating the expres-sion of miRNAs such as miR-133a-5p, miR-133b-5p, miR-664-1-5p, miR-6216 and let-7e-5p.
9.Association of serum C peptide level with the severity of diabetic foot ulcers and its healing rate
Shumin WANG ; Yang HE ; Lei XU ; Kai GUO ; Junyi GU ; Yaping SHEN ; Xueming GU ; Zhengyi TANG
Chinese Journal of Endocrinology and Metabolism 2017;33(1):17-22
Objective To investigate the association of serum C peptide concentration with the severity and the outcome of diabetic foot ulcer (DFU). Methods The clinical data of 257 inpatients with DFU were collected, including fasting and postprandial 2h C peptide levels and C peptide area under curve (AUCCP ). The patients were followed up on the outcomes of ulcers and death. The associations of serum C peptide concentration with the Wagner degree, infection severity, and healing rate were analyzed. Results The medians of fasting and 2h postprandial serum C peptide as well as AUCCP were 1. 37(0. 02 ~ 9. 00) nmol/ L, 3. 22(0. 02 ~ 29. 61) nmol/ L, and 511. 65 (3. 60 ~ 2 691. 30)nmol·min-1 ·L-1 respectively, which were lower than general levels. The time of follow-up in our study was 2. 8 (1. 0 ~ 5. 1) years. By the end of study, the wound of 75. 88% patients was healed, 3. 5%undergone major amputation, and 23. 74% died. After adjusting for relative factors, there were no significant associations of serum fasting and postprandial C peptide levels and AUCCP with Wagner degree and infection severity (P>0. 05). Cox regression analysis showed that the fasting plasma C peptide and hemoglobin were the independent protective factors for the healing of ulcers; old age, male, higher infection degree, and diabetes family history were their independent risk factors ( all P < 0. 05). Conclusions The lower plasma fasting C peptide concentration in patients with DFU is not correlated with Wagner degree and infection severity, but closely related with healing rate.
10.Exosomes serected by mesenchymal stem cells derived form orofacial bone regulate the function of macrophage
Lin YUAN ; Yina CAO ; Zhengyi YANG ; Jin SUN ; Guangsi PAN ; Jun QIAN ; Jingjing SONG ; Han WANG
Journal of Practical Stomatology 2017;33(3):344-348
Objective:To investigate the trait of exosomes serected by mesenchymal stem cells derived form orofacial bone(OMMSCs-Exo) and the communication between the exosomes and macrophages.Methods:OMMSCs were isolated from orthognathic surgical sites and cultured by limited dilution.Their cell surface markers were characterized by flow cytometry.the rate of colony formation and the differentiation potential of OMMSCs were evaluated.Exosomes were prepaired from the culture supernatants of OMMSCs(P4-P6).Transmission electron microscopy(TEM) and western blot were used to identify the exosomes.The expression of miRNAs associated with immunity such as miR-223 and miR-let-7c were determined by Real-time RT-PCR.Human peripheral blood mononuclear cells(PBMCs) were isolated from health donor and co-cultured with OMMSCs-Exo.After co-cultured for 24 h,the communication between exosomes and macrophages was tested using a confocal microscope.Results:Human OMMSCs were proved to have the characteristics of mesenchymal stem cells.The diameter of OMMSCs-Exo ranged from 40 to 160 nm.The OMMSCs-Exo expressed CD63 and CD81 and contained miRNAs associated with immune regulation such as miR223 and miR-let-7c.OMMSCs-Exo could be uptaken by macrophages.After co-culture of OMMSCs-Exo with marcrophages for 72 h,miR223 expression in macrophages increased.Conclusion:OMMSCs-Exo has the potential of immune regulation.