1.Improvement effects of astragaloside Ⅳ on myocardial focal ischemia-reperfusion inj ury and its influence in PI3K/Akt/mTOR signaling pathway
Jing ZHANG ; Cuili MA ; Zhiguo WANG
Journal of Jilin University(Medicine Edition) 2014;(5):991-996
Objective To observe the improvement effects of astragaloside Ⅳ (AS-Ⅳ)on the myocardial focal ischemia-reperfusion (I/R)injury and its influence in PI3K/Akt/mTOR pathway,and to clarify the protective effect of AS-Ⅳ on myocardial I/R inj ury and the possible mechanisms.Methods The left main coronary arteries of 60 Sprague-Dawley rats were occluded for 30 min followed by a 120-min reperfusion to induce I/R model.The rats with I/R inj ury were randomly divided into model group (normal saline), AS-Ⅳ group (intravenous inj ection of 10 mg·kg-1 AS-Ⅳ 5 min before reperfusion),PI3K inhibitor Wortmannin (WOR)group (intravenous injection of 0.6 mg·kg-1 WOR 10 min before reperfusion)and AS-Ⅳ+WOR group (intravenous injection of 10 mg·kg-1 AS-Ⅳ and 0.6 mg·kg-1 WOR 5 and 10 min before reperfusion,respectively).15 age-matched SD rats were chosen as control group.The heart mass,degrees of infarction and ischemia and cardiac function ,including left ventricular systolic mean pressure (LVSP),end-diastolic pressure (LVEDP),fractional shortening (FS)and ej ection fraction (EF),of the rats in all groups were analyzed. Western blotting method was used to measure the phosphorylation levels of Akt and mTOR(p-Akt and p-mTOR).The specific fluorescent probe DHE staining was employed to detect the myocardial reactive oxygen species levels. Results Compared with control group, the degrees of infarction and ischemia, LVEDP, myocardial levels of p-Akt/Akt, p-mTOR/mTOR and reactive oxygen species levels of the rats were increased (P<0.05).and the levels of LVSP,FS and EF were decreased in model group (P<0.05). Compared with the model group,the degrees of infarction and ischemia,LVEDP and reactive oxygen species level were decreased (P<0.05),while the levels of p-Akt/Akt,p-mTOR/mTOR LVSP, FS and EF of all rats in AS-Ⅳ group were increased (P<0.05).Compared with AS-Ⅳ group,the degrees of infarction and ischemia,LVEDP and reactive oxygen species levels of the rats in WOR group and AS-Ⅳ+WOR group were increased (P<0.05 ), and the myocardial p-Akt/Akt, p-mTOR/mTOR and LVSP, FS, EF were decreased (P<0.05).Conclusion AS-Ⅳ has improvement effect on myocardial I/R injury.AS-Ⅳ can reduce the extent of myocardial infarction and oxidative stress and improve the heart function,and its possible mechanism may be related to activating PI3K/Akt/mTOR signaling pathway.
2.Drug-release effect of fluorouracil implants in protein denaturant hydrochloric acid
Shiliang WANG ; Jing WANG ; Qingsheng YIN ; Cuili REN ; Xiaoqin MA ; Mei FENG
Chinese Journal of Tissue Engineering Research 2009;13(12):2395-2400
BACKGROUND: Directly percutaneous injection of protein-denaturant hydrochloric acid (PDHA) into tumors can lead to fast killing of tumor, sustained drug release and prevention of in situ recurrence of tumor. However, whether implants can be used combined with denaturant still remains unknown. OBJECTIVE: To investigate the compatibility of fluorouracil implants and PDHA (6 mol/L). DESIGN, TIME AND SETTING: Observational study was performed in the Hefei Industry University between October 2006 and March 2007. MATERIALS: A total of 78 Wistar rats, weighing (200i20) g, half males and half females, were used for testing drug release in vivo. Drugs fluorouracil implants (H20030345; columniform particle, diameter 0.8 mm, length 4 mm; specifications: Fluorouracil 2 mg/particle; batch number: 20060922; meeting the National Drug Quality Standards [WS1-(X-103)-2005Z]) were provided by Wuhu Zhongren Pharmaceutical Company,Ltd. Hydrochloric acid (37%) was analytical reagent. METHODS: 96 tubes of the implants and PDHA were kept at (37.0± 0.5) ℃. Each time, six samples were collected at 1, 8, 16, 24, 96, 120, 168, 240, 360, 432, 480, 528, 600, 720, and 960 hours after incubation. Appearance of the implants was observed by microscope. Stability of fluorouracil in PDHA was determined by HPLC and ultraviolet absorb method. Based on the entering quantity and residual quantity of fluorouracil, the release rates were calculated. MAIN OUTCOME MEASURES: The approximate solubility, stability and morphological change of fluorouracil in denaturant and the corresponding drug release character in both denaturant and rats in vivo. RESULTS: At (37.0±0,5) ℃, the fluorouracil was stable for 960 hours in PDHA, the saturated concentration of fluorouracil was (22.72±0.04) g/L. The appearance of implants was intact. The surface was porous. Compared with the speed of releasing drug in rats, the speed of releasing drug was faster in the early stage of release process and slower in the later stage. The drug release was incomplete. At 1, 24, 96, 360 and 960 hours, the implants' release rates were (11.9±6.7)%, (37.9±5.3)%, (52.6±4.5)%, (75.3±3.8)%, and (85.5±2.1)%, respectively. CONCLUSION: The fluorouracil implants and hydrochloric acid (6 mol/L) are compatible and no influence is detected during the observation.
3.The immune efficacy of OmpH DNA vaccine from avian Pasteurella multocida
Qiang GONG ; Mingfu NIU ; Shuaitao WANG ; Cuili QIN ; Xiaofei SUN ; Liping MA ; Yuze HOU
Chinese Journal of Immunology 1985;0(01):-
Objective:To research on protective immunity of omph DNA vaccine against avian Pasteurella multocida in mice.Methods:The omph gene fragment amplified by PCR from avian Pasteurella multocida was cloned into pMD18-T.Subsequently it was subcloned into the eukaryotic expression vector pcDNA3.1(+),and the recombinant plasmid pOMPH was obtained.Then the recombinant plasmid was transfected into SP2/0 cells in vitro.The transcription and expression of target gene were analyzed by RT-PCR,Western blot analysis and indirect immunofluorescence.Three groups of BALB/c mice(n=16) named pOMPH,pCDNA3.1(+) and PBS were intramuscularly vaccinated with the recombinant plasmid,control vector and PBS respectively.The serum antibodies were detected by indirect ELISA.The spleen lymphocyte proliferation (SLP) and secreted IFN-? of spleen were tested by MTT.The mice were challenged with virulent of avian Pasteurella multocida on week 2 post the third immunization,the protection rate were counted.Results:RT-PCR,Western blot analysis and indirect immunofluorescence showed that the omph gene could be transfected into SP2/0 cells in vitro and expressed the target protein.Indirect ELISA showed that the levels of antibodies in pOMPH group were most significantly higher than in the other groups(P
4.The immune efficacy of Th/H DNA vaccine from avian Pasteurella multocida
Qiang GONG ; Mingfu NIU ; Shuaitao WANG ; Cuili QIN ; Xiaofei SUN ; Liping MA ; Yuze HOU
Chinese Journal of Immunology 2010;26(1):13-16,22
Objective:To research on protective immunity of omph DNA vaccine against avian Pasteurella multocida in mice.Methods: The omph gene fragment amplified by PCR from avian Pasteurella multocida was cloned into pMD18-T.Subsequently it was subcloned into the eukaryotic expression vector pcDNA3.1(+),and the recombinant plasmid pOMPH was obtained.Then the recombinant plasmid was trans fected into SP2/O cells in vitro.The transcription and expression of target gene were analyzed by RT-PCR,Westem blot analysis and indirect immunofluorescence.Three groups of BALB/c mice(n=16) named pOMPH,pCDNA3.1(+) and PBS were intramuscularly vaccinated with the recombinant plasmid,control vector and PBS respectively.The serum antibodies were detected by indirect ELISA.The spleen lymphocyte proliferation (SLP) and secreted IFN-γof spleen were tested by MTT.The mice were challenged with virulent of avian Pasteurella multocida on week 2 post the third immunization,the protection rate were counted.Results: RT-PCR,Western blot analysis and indirect immunofluorescence showed that the omph gene could be,transfected into SP2/0 cells in vitro and expressed the target protein.Indirect ELISA showed that the levels of antibodies in pOMPH group were most significantly higher than in the other groups(P<0.01).Spleen lymphocyte proliferation by MTT assay indicated that the SI value induced with avian Pasteurella multocida Omps in pOMPH group was higher than those in pCDNA3.1 (+) and PBS groups (P<0.05).The IFN-γexperiments(Double-antibodies-sandwich-ELISA)showed that the levels of IFN-γ induced with Omps in the group of pOMPH was mostly higher than in the other control groups apperent(P<0.01 ).The protection rate of pOMPH(70%) was better than in the other groups.Conclusion: The omph DNA vaccine against avian Pasteurella multocida had been constructed successfully.The DNA vaccine could enhance the immunity level and the protective effect of the vaccinated mice.Present study may be useful for the development of avian Pasteurella multocida vaccine.
5.Research advances in chronic inflammatory demyelinating polyradiculoneuropathy
Chinese Journal of Neurology 2023;56(2):233-240
Chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) is a chronic autoimmune disease affecting the peripheral nervous system mediated by cellular and humoral immunity, characterized by limb weakness and sensory impairment. The main feature of CIDP by electrophysiological and pathological examinations is the demyelination of peripheral nerves. First-line treatment for CIDP includes glucocorticoids, intravenous immunoglobulins, and plasmapheresis. Some patients respond to current treatment not well and have a poor prognosis. Progress in the pathogenesis, diagnosis, and treatment of CIDP worldwide was reviewed in this article, aiming to provide references for the clinical diagnosis and treatment of CIDP.
6.Construction of a predictive model based on columnar maps for factors influencing lesion residuals after minimally invasive rotary mastectomy for breast masses
Changxin LYU ; Zejie SHAO ; Li LI ; Liyong FENG ; Shengjie MA ; Cuili LIU
Chinese Journal of Endocrine Surgery 2023;17(1):84-89
Objective:To investigate the influencing factors associated with lesion residual after minimally invasive rotary mastectomy for breast masses and to construct a predictive model using columnar plots.Methods:Two hundred and twenty-eight patients with Breast lumps in Linyi People’s Hospital from Jun. 2018 to Jun. 2020 were selected as study subjects, all of whom underwent minimally invasive rotational resection, and the influencing factors of postoperative lesion residual were analyzed using univariate and multifactorial analysis, and a column line graph risk warning model was constructed and given for evaluation and validation.Results:228 patients were followed up for 6 months after surgery, 3 cases were lost, 225 cases completed postoperative follow-up, among which 185 cases (82.22%) had successfully resected lesions; 40 cases (17.78%) had residual lesions. Univariate, logistic regression analysis showed that tumor diameter ( t=15.52, P<0.001) , lesion morphology ( t=15.52, P<0.001) , lesion boundary ( χ2=7.71, P=0.006) , ultrasound guidance ( χ2=12.69, P<0.001) , and minimally invasive spinotomy system ( χ2=10.64, P=0.001) were the factors influencing lesion residual after minimally invasive spinotomy of breast masses. Based on the above factors to construct a column line graph model of lesion residual after minimally invasive rotational mastectomy for breast lumps, the area under the working characteristic curve (ROC) for model subjects was 0.974 (95% CI: 0.954-0.994) , indicating that the calibration curve basically matched the actual curve; external validation of the model showed that the area under the obtained ROC was 0.962 (95% CI: 0.933-0.991) , indicating that the model has a high degree of calibration. Conclusion:Tumor diameter, lesion morphology, lesion boundary, ultrasound guidance, and minimally invasive spinotomy system are the influencing factors of lesion residual after minimally invasive spinotomy for Breast lumps, and the column line graph model prediction of lesion residual occurring after surgery constructed based on the above factors has good confidence.