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The experimental class is a very important part of teaching process for medical students. Relying on the medical virtual simulation experiment center of Henan University, in this study, we select the appropriate experimental items and related knowledge content of the medical virtual simulation experiment center and post them to the students. Students are required to log in to the experimental center to strengthen the study of basic medical experimental knowledge by watching experimental videos and learning experimental principles, and then carry out virtual experimental operations. In terms of the teaching content, the required experimental virtual operation are required to complete, and in terms of teaching form, we adopt the combination of online experimental teaching course and webcast course teaching. This experimental teaching method is easy for students to accept, stimulates high enthusiasm for learning, and inspires curiosity for basic medical knowledge for the students, which significantly improves the teaching effect.
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Objective:To explore the role of interleukin (IL)-9 and other T helper (Th) cell-related cytokines in the pathogenesis of acute anterior uveitis.Methods:A cross-sectional study was conducted.Thirty-six patients (36 eyes) with acute anterior uveitis who were treated at Gansu Provincial Hospital from May 2018 to May 2019 and 40 matched healthy subjects (40 eyes) who had no eye diseases or systemic diseases in the same period were enrolled as the acute anterior uveitis group and healthy control group, respectively.The disease severity of the subjects in the acute anterior uveitis group was graded and the subjects were divided into mild, moderate and severe groups according to the grading.Serum of all subjects was collected to determine the concentration of serum IL-9, IL-17, transforming growth factor-β 1 (TGF-β 1), interferon-γ (IFN-γ), IL-4, IL-35 and IL-22 by enzyme linked immunosorbent assay (ELISA) method.Spearman rank correlation analysis was used to evaluate the relationship between IL-9 concentration and other Th cell-related cytokines.This study adhered to the Declaration of Helsinki.The study protocol was approved by an Ethics Committee of Gansu Provincial Hospital (No.2019-204). Written informed consent was obtained from each subject prior to any medical examination. Results:The serum levels of IL-9, IFN-γ, IL-4, TGF-β 1, IL-35 and IL-22 in the acute anterior uveitis group were significantly higher than those in the healthy control group, and the differences were statistically significant (all at P<0.05). There was no statistically significant difference in the concentration of IL-17 between the two groups ( U=704.500, P=0.872). The IL-9 concentration of patients with acute anterior uveitis in the mild, moderate and severe groups was 57.24 (47.47, 65.10), 71.68 (67.55, 78.91) and 114.01 (74.78, 139.30) ng/L, respectively, and the overall difference was statistically significant ( Z=8.766, P=0.012), and the IL-9 concentration of the mild group and the moderate group was significantly lower than that of the severe group (both at P<0.05). The concentration of IL-9 in the patients with acute anterior uveitis was positively correlated with the concentration of IL-17, TGF-β 1 and IL-35 ( rs=0.449, 0.517, 0.400; all at P<0.05), and no significant correlations were found between the concentration of IL-9 and the concentration of IFN-γ, IL-4 and IL-22 ( rs=0.293, 0.286, 0.316; all at P>0.05). Conclusions:IL-9 plays a role in promoting the immune inflammatory response in the occurrence and development of acute anterior uveitis, and it is closely related to Th17 and Treg cell-related cytokines (TGF-β 1, IL-35).
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Objective:To determine the effects of Foxp3-overexpressing lung cancer cells on activated CD4+T lymphocyte.Methods: Stable Foxp3-overexpressing lung cancer cells NCIH-1299,NCIH-hFoxp3,was generated by transfection of NCIH-1299 cells with plasmid pcDNA3-hFoxp3 mediated by Lipofectamine 2000 and by selection with G418,and validated by quantitative PCR and Western blot.The expression levels of IL-8 and IL-10 secreted by NCIH-hFoxp3 and NCIH-control were measured by ELISA.IL-2 secrection by activated human CD4+T lymphocyte which was tested after stimulation with 20% conditioned medium of NCIH-hFoxp3 and NCIH-control cells.The proliferation of activated human CD4+ T lymphocytes was assessed by MTT after coculture with NCIH-hFoxp3 cells.The adhesive ability of activated human CD4+ T lymphocytes was probed with NCIH-hFoxp3 cells by immunocytochemistry.Results: Compared with NCIH-control cells,NCIH-hFoxp3 secreted high level of IL-10 and low level of IL-8.NCIH-hFoxp3 with Foxp3 overexpression significantly suppressed the proliferation,adhesive potential and IL-2 expression by activated CD4+ T cells.Conclusion: Suppression of immune activities of activated CD4+ T cells by Foxp3 overexpression in lung cancer cells may correlate with cytokine IL-8 and IL-10,which can contribute lung cancer progression.
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Objective:To determine the effects of enforced expression of Foxp3 in lung cancer cell with regards to proliferation and tumorgeneity. Methods: A stable subline NCIH-hFoxp3 was established by liopfectamin-mediated pcDNA plasmid transfection carrying exogenous hFoxp3. The growth curve and secrection of IL-8 and IL-10 of NCIH-hFoxp3 were evaluated using MTT and ELISA, respectively. The in vivo tumorigeneity was assessed as well by inoculation of NCIH-hFoxp3 subcutaneously in nude mice. Results:Lung cancer cell NCIH-hFoxp3 with enforced expression of Foxp3 proliferated slowly but exihited increased in vivo tumorgeneity compared with corresponding control subline. In addition,increased expression of hFoxp3 in NCIH-hFoxp3 augmented secretion and at-tenuated secretion of IL-8 and IL-10,respectively. Conclusion:Increased expression of Foxp3 may promote progression of lung cancer cell by change of cellular microenvironment and evasion of immune surveillance.
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Objective To observe the expression and mechanism of hypoxia-inducible factor-1α (HIF-1α) and p53 protein at the altitude of 5000 meter plateau hypoxia environment in rats,as well as the effect of Astragalus injection.Methods Sixty Sprague Dawley rats were randomly divided into the Astragalus injection intervention group and normal saline control group,30 rats in each group.Astragalus injection group rats were intraperitoneal injected of Astragalus injection (15 ml/kg) before 30 minutes into the plateau environment simulation cabin,normal saline group rats were intraperitoneal injected with the same volume of saline.30 minutes after injection,rats in each group were reared in the plateau experiment cabin which simulated altitude of 5000 m (oxygen partial pressure 11.3 kPa) for 2,6,8,12,24 hours,each time period of 6 rats.When get out,the rats were executed immediately and eyes were harvested.Retinal sections were studied by hematoxylin eosin stain,and immunohistochemical method for HIF-1α and p53 expression.Results For control rats,after 2 hours in the cabin,there was edema in retinal layers.HIF-1α and p53 were expressed mainly in the cytoplasm of retinal layers.When the periods in cabin extended,there was atrophy of retinal nerve fiber layer,swelling and degeneration of ganglion cells.The expression of HIF-1α and p53 was increased.Compared with the control group,the intervention group rat had similar but less severe retinal changes,and the expression of HIF-1α and p53 was significantly decreased (P ＜0.05).Conclusion Astragalus injection can reduce pathological retinal damage in rats at high altitude environment,and its mechanism may be associated with reduced HIF-1α,p53 expression.
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Background The simplex congenital blepharoptosis is the common blepharon motor dysfunction disease.Some researches have shown that congenital blepharoptosis is related to the hypoplasia of levator.Objective This study was to investigate the thickness and pathological features of levator palpebrae superioris aponeurosis in congenital blepharoptosis patients.Methods A prospective cohort study was carried out in Anyang Eye Hospital from March 2012 to April 2014.Eighty-five eyes of 56 patients with congenital blepharoptosis were divided into mild (15 eyes), moderate (25 eyes) and severe blepharoptosis (19 eyes) groups, and the fellow eyes of monocular blepharoptosis was used as fellow eye group (26 eyes).Twenty-six eyes of 13 normal subjects were recruited for the normal control group.The thickness of levator aponeurosis was measured by ultrasound biomicroscope (UBM) , and the shifting range of levator aponeurosis was detected by using measuring scale.Levator aponeurosis specimens were collected during the levator palpebrae superioris shortening surgery for the pathological examination.The study was approved by the medical ethics committee of Anyang Eye Hospital, and the patients or their guardian signed the informed consent.Results The thickness of levator aponeurosis was (0.331±0.018), (0.373±0.026), (0.539± 0.023) , (0.557 ± 0.024) and (0.547 ± 0.028) mm in the severe blepharoptosis group, moderate blepharoptosis group,mild blepharoptosis group, normal control group and fellow eye group, respectively, showing a significant difference among them (F =1.681, P =0.043).The thickness values of levator aponeurosis were considerably lower in the severe blepharoptosis group and moderate blepharoptosis group than those in the mild blepharoptosis group,fellow eye group and normal control group (all at P＜0.05) , and the thickness value of levator aponeurosis was significantly reduced in the severe blepharoptosis group compared with the moderate blepharoptosis group (P＜0.05).Pathological examination showed arranging disorder of muscle fibers,hyaline-like degeneration, connective tissue hyperplasia and interruption of endomysium.The number of eyes with severe hyaline-like degeneration and connective tissue hyperplasia was significantly increased in the severe blepharoptosis group than that in the moderate blepharoptosis group or the mild blepharoptosis group, as well as in the moderate blepharoptosis group than that in the mild blepharoptosis group(all at P＜0.01).The adipose cells in muscle in the mild blepharoptosis group, moderate blepharoptosis group and severe blepharoptosis group were (12.35±4.62), (17.58±7.46) and (26.19±10.81)/field,and adipose cells in the severe blepharoptosis group were significantly more than those in the mild and moderate blepharoptosis groups (t =5.60, P =0.00;t =2.71, P =0.01).A significant increase in the adipose cells also was seen in the moderate blepharoptosis group compared with the mild blepharoptosis group (t =2.44, P =0.02).Conclusions UBM can offer accurate thickness data of levator aponeurosis.The combination of thickness data and shifting range measurement of levator aponeurosis is helpful for the evaluation of muscle strength.The development of levator aponeurosis appears to be abnormal in congenital blepharoptosis patients.The histopathological change parallels to the severity of the disease.
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Objective:To determine the correlation of BCG concentration in Freund complete adjuvant ( FCA) and severity of arthritis during arthritis establishment in DBA 1/j male mice induced by bovine type Ⅱcollagen.Methods:CIA was induced by the im-munization of DBA1/j mice with bovine type Ⅱcollagen and BCG of various concentrations dissolved in FCA.To ascertain the effects administering the collagen booster,CIA-related features(including body weight,clinical scoring of arthritis),TNF-αin serum and the histopathological changes in the spleen and the joints regions were measured.Results:4 mg/ml BCG induced more serious arthritis than 1 mg/ml in DBA1/j mice after collagen exposure.Conclusion:There is a positive correlation of arthritis severity with BCG.Which indicates that selection of adjuvant with suitable BCG concentration could determine pathological outcome in CIA mouse model .
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Objective To investigate the expression of TIPE2 in splenic lymphocytes derived from the mouse model of cattle collagen Ⅱ-induced rheumatoid arthritis (CIA) and from peripheral blood of patients with rheumatoid arthritis (RA) for its role in the pathogenesis of RA.Methods The CIA mouse model was established by immunization of DBA-1/J mice with cattle collagen Ⅱ.Peripheral blood lymphocytes were collected from RA patients and healthy donors.The TIPE2 mRNA and protein expression in splenocytes of CIA and control mice were measured by semi-quantitative RT-PCR and Western blot,respectively.In addition,fluorescence quantitative RT-PCR and Western blot were used to determine the TIPE2 mRNA and protein expression in peripheral blood lymphocytes derived from patients with RA and healthy donors.Results Both mRNA and protein expression of TIPE2 were higher in splenocytes of CIA mice and lymphocytes of peripheral blood from RA patients compared with corresponding controls.Furthermore,the mRNA and protein expression of TIPE2 increased significantly in patients with active RA.TIPE2 expression were positively correlated with DAS28 scores (P＜0.001).Conclusion Our results suggest that TIPE2 plays a role in the RA pathogenesis.The level of TIPE2 may also indicate the severity of rheumatoid arthritis.
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The attempt of vitrectomy is to remove the lesion of vitreous body and relieve vitreous traction to retina.Whether the vitrectomy is successful or not depend on if exist a close adhesion between vitreous body and retina or a posterior vitreous detachment (PVD).The drug assisted vitrectomy is a new technique.These drugs can help surgeon to form PVD during surgery safely and effectively.The drug is utilized prior to vitreoretinal operation to promote the colliquate of vitreous body or dissolve of fibroplasia membrane of the vitreous body in order to release the traction between the posterior cortex of vitreous body and the inner limiting membrane of retina.The research advance in relevant drugs is reviewed in this article.
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Objective:To investigate mitochondrial-dependent molecular mechanisms of a novel agonistic anti-human death receptor 5 (DR5) monoclonal antibody(mDRA-6) inducing apoptosis in Jurkat cell.Methods:The dose-dependent and time-dependent cell growth suppression of mDRA-6 in Jurkat cells was determined by MTT assay.The measurement of the mitochondrial transmembrane potential(ΔΨm) of Jurkat cells was detected by flow cytometry with JC-1 single staining.Caspase-8,9 as well as Bid,Bax,Bcl-2 and Cyto c of apoptotic Jurkat cells were analyzed by Western blot after mDRA-6 treatment.Results:The mDRA-6 induced cell growth suppression and cytotoxicity in dose-dependent manner and time-dependent manner.After mDRA-6 treatment at 2.0 μg/ml for15 min,30 min,60 min and 120 min,the change in ΔΨm were 20.14 %,19.34 %,21.11% and 30.90% respectively by JC-1 single staining.Western blot revealed that the level of active fragments of Caspase-8,9 and Bid,Bax,Bcl-2 and Cyto c respectively,and the amount of Cyto c was increased in cytosol concomitant with the related attenuation of Cyto c in mitochondria.Conclusion: Apoptotic pathway of Jurkat cells induced by mDRA-6 is initiated upon DR5 ligatian to mDRA-6 and exogenic Caspase-dependent cell apoptotic cascades is activated,and endogenic mitochondrial-dependent cell apoptosis pathway is activated.mDRA-6 may be a useful agent in investigating human leukemia therapy by using TRAIL/DR5.
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Objective To explore the effect of TNF related apoptosis inducing ligand (TRAIL) in apoptosis induced by LPS. Methods After LPS injected mice blocking TRAIL with soluble death receptor 5 (sDRS), detecting ALT, AST and LDH of mice serum at different times, apoptotic effects of LPS to mice hepatocyte were detected by HE and flow eytometry (FCM) with Annexin V-FITC/PI staining. The expres-sion of DR5 in mice hepatocyte was assayed with immunohistochemistry and FCM. Results Apoptotic effect was promoted by up-regulated DR5 expression on hepatocyte. Blocking TRAIL with sDR5 markedly amelio-rated the hepatocyte damage and reduced apoptosis. Conclusion These results establish a critical role for TRAIL in apoptosis during disease process of LPS.
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Objective:To observe anti-DR5 monoclonal antibody on apoptosis and CDC effect in EC109 cells.Methods:The anti-DR5 monoclonal antibody was prepared by hybridoma technique.Tumoricidal effects and complement-dependent cytotoxicity of the McAb to EC109 cells was screened by MTT assay.The apoptosis of EC109 cells was detected by flow cytometry with annexin Ⅴ-FITC/PI staining.Morphological change of EC109 was observed by microphotograph.Results:An anti-DR5 monoclonal antibody was obtained.It induced apoptosis of EC109 dose dependently.The cytotoxic action was notably enhanced by addition of complement.The cells growth inhibition ratios reached 83.04%.The apoptotic body and cathepsis were seen in microphotograph.Conclusion:The anti-DR5 monoclonal antibody could induce EC109 cells apoptosis and cause the complement dependent cytotoxic (CDC) effects powerfully.