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Objective To investigate the cytoprotection and mechanism of carbachol(CCH)to stimulate M3mus-carinic acetylcholine receptor(M3-mAChR) against hypoxia injury induced by cobaltous chloride hexahydrate (CoCl2) in rat cardiomyocyte line H9c2. Methods Select the normal rat cardiomyocyte line H9c2 as the control group, rat cardiomyocyte line H9c2 was managed with CoCl2to develop hypoxia injury model, M3-mAChR spe-cific agonist CCH and antagonist 4-diphenyl-acetoxy-N-methyl-piperidine methiodide(4-DAMP) were used for in-tervention. The cell viability was tested by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT); The apoptosis in cardiomyocyte was detected by flow cytomery(FCM); The expression of M3-mAChR,caspase-3, HIF-1α and HO-1 proteins was measured by Western blot assay. Results In hypoxia group,the ap-optosis rate was significantly increased while cell proliferation decreased, the expression of HIF-1α, caspase-3 and HO-1 proteins were up-regulated obviously;After treatment with CCH,the apoptosis and cell proliferation of cardiomyocytes were significantly decreased, while the proliferation of cells increased, and the expression of M3-mAChR, HIF-1α and HO-1 proteins increased, the expression of caspase-3 protein was significantly decreased. Moreover, when applying 4-DAMP as intervention, these effects mediated by CCH was abolished.Conclusions CCH stimulates M3-mAChR against hypoxia injury induced by CoCl2in rat cardiomyocyte strain H9c2, and the mechanism may be related to down-regulation of caspase-3 expression and up-regulation of HIF-1α and HO-1 protein expression.
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Objective To investigate the clinical value of total hysterectomy in the treatment of cervical intraepithelial neoplasia. Methods We retrospectively analyzed the clinicaldata of 283 patients undergoing total hysterectomy for cervical intraepithelial neoplasia in Changhii Hospital of Second Military Medical University from Jan. 2009 to Dec. 2016. Among the patients, 206 cases received loop electrosurgical excision procedure CLEEP) before total hysterectomy (pre-operative LEEP group), 28 only received total hysterectomy directly after colposcopy biopsy (hysterectomy group) and 49 received LEEP during total hysterectomy (intra-operative LEEP group). The changes of pathological grade and the lesion residual rates of the patients before and after operation were compared. The effect of clinicopathological features (age, pregnancy times, partus times, abortion times, menopausal years, pathology, human papilloma virus (HPV) infection, tumor family history and so on) on postoperative residual lesion were analyzed by univariate and multivariate analysis. Results The cure rate, lesionresidual rate, pathological downgrade rate and pathological upgrade rate in the pre-operative LEEP group was 35. 92%(74/206), 64. 08%(132/206), 63. 11% (130/ 206) and 3. 88% (8/206), respectively, and two cases with canceration were found. In the hysterectomy group, the consistent rate of pathological diagnosis before and after operation, pathological downgrade rate and pathological upgrade rate were 57. 14%(16/28), 35. 71 %(10/28) and 7 14%(2/28), respectively, and one case with invasive carcinoma was found after operation. The lesion residual rate, pathological downgrading rate and pathologicil upgrading rate in the intra- operative LEEP group were 40. 82% (20/49), 65. 31% (32/49) and 8. 16% (4/49), respectively, and two cases with invasive carcinoma were found after operation. Univariate analysis showed that age and menopausal years were the factors influencing lesion remaining after LEEP (P<0. 05). Multivariate analysis showed that patients with longer menopausal years had lower risk of lesion remaining (P = 0. 02). The pregnancy times, partus times, abortion times, lesion involving glands, positive margins, HPV infection and family history of cancer had no significant effects on postoperative lesion remaining. Conclusion Total hysterectomy plays an important role in preventing the recurrence of cervical precancerous lesion and reducing the incidence of invasive carcinoma. It is suitable for people with few screening opportunities, poor follow-up conditions and no fertility requirements. Patients with high grade intraepithelial lesion who are not menopausalor have short menopausal years need to be followed up more closely and can receive total hysterectomy if necessary.
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In order to assess the number and function of macrophages in the placenta of pregnancy complicated with gestational diabetes mellitus (GDM) as well as those of normal pregnancies, placenta samples were collected from 15 GDM patients (GDM group) and 10 normal pregnant women (control group). The expression levels of macrophage markers (CD68/CD14) and inflammatory cytokines (IL-6/TNF-α) in placenta were detected using immunohistochemistry and PCR. The results showed that the number of CD68+ or CD14+ cells in the GMD group was remarkably higher than that in the control group (P<0.05), indicating that the number of macrophages in the GDM group was significantly greater than that in the control group. The mRNA expression levels of CD68+, IL-6 and TNF-α were higher in the GMD group than in the control group. In conclusion, more macrophages accumulate in placenta of pregnancy complicated with GDM, and the expression levels of pro-inflammation factors are also increased in GDM pregnancies, suggesting that macrophages and inflammatory mediators (IL-6 and TNF-α) may play an important role in GDM.
Subject(s)
Adult , Female , Humans , Pregnancy , Antigens, CD , Antigens, Differentiation, Myelomonocytic , Cell Count , Cytokines , Genetics , Allergy and Immunology , Metabolism , Diabetes, Gestational , Genetics , Allergy and Immunology , Metabolism , Immunohistochemistry , Inflammation Mediators , Allergy and Immunology , Metabolism , Interleukin-6 , Genetics , Allergy and Immunology , Metabolism , Lipopolysaccharide Receptors , Genetics , Metabolism , Macrophages , Allergy and Immunology , Metabolism , Pathology , Placenta , Allergy and Immunology , Metabolism , Reverse Transcriptase Polymerase Chain Reaction , Tumor Necrosis Factor-alpha , Genetics , Allergy and Immunology , MetabolismABSTRACT
In order to assess the number and function of macrophages in the placenta of pregnancy complicated with gestational diabetes mellitus (GDM) as well as those of normal pregnancies, placenta samples were collected from 15 GDM patients (GDM group) and 10 normal pregnant women (control group). The expression levels of macrophage markers (CD68/CD14) and inflammatory cytokines (IL-6/TNF-α) in placenta were detected using immunohistochemistry and PCR. The results showed that the number of CD68+ or CD14+ cells in the GMD group was remarkably higher than that in the control group (P<0.05), indicating that the number of macrophages in the GDM group was significantly greater than that in the control group. The mRNA expression levels of CD68+, IL-6 and TNF-α were higher in the GMD group than in the control group. In conclusion, more macrophages accumulate in placenta of pregnancy complicated with GDM, and the expression levels of pro-inflammation factors are also increased in GDM pregnancies, suggesting that macrophages and inflammatory mediators (IL-6 and TNF-α) may play an important role in GDM.
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Objective:To study the influence of morphine on the expression of nestin in the ependymal epithelia,central gray and hippocampal formations in mice.Methods:Twenty health mice were evenly randomized into control group and experiment group.Mice in the control group were injected with normal saline(0.1 ml daily)and those in the experimental group were injected with morphine (0.1 ml,1 mg daily).Thirty days later,the mice brain samples were harvested and made into paraffin sections.Immumohistochemical ABC technique was used to observe the expression of nestin under light microscope.The images were analyzed with the image analytical system.Results:In the control group,the ependymal epithelia,the central gray,the periventricular gray substances and the hippocampal formations had weak expression of the nestin,with a mean gray scale of 150.98?13.31;there were 5 kinds of nestin-positive cells:(1) the basal cells of ependymal epithelium,(2)cells distributed in the periventricular gray substance and the deep lamella of central gray, (3)cells distributed in the superficial lamella of central gray,the subiculum,the parahippocampal gyrus and the cortex inⅡ,Ⅲlayers of the entorhinal area,(4)cells frequently seen in the rectum of midbrain and the subiculum,and(5)cells distributed in the tectum of midhrain,the hippocampus,gyrus dentatus,parahippocampal gyrus and the cortex in V layer of the entorhinal area;the density of nestin in the subiculum and entorhinal area was(7.20?1.23)mm~2.In the experiment group,the ependymal epithelia,the central gray,the periventricular gray substances and the hippocampal formations had positive expression of the nestin,with the mean gray scale being 133.03?22.28;the density of the above-mentioned 5 kinds of cells increased;the density of nestin in the subiculum and entorhinal area was(10.50?1.43)mm~2.The mean value of gray scale and nestin-positive neurons were significantly different between the 2 groups (P
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Objective:To study the influence of morphine on the expression of synaptophysin(SYN)and synapse structure in mice hippocampus,so as to provide pathological evidence for studying the development and treatment of chronic morphine intoxication, addiction and abstinence symptoms of morphine.Methods:Twenty mice were evenly randomized into control group and experiment group.Mice in control group were injected with normal saline(0.1 ml daily)and those in experimental group were injected with morphine(0.1 ml,1 mg daily).Thirty days later the mice in 2 groups were killed and their brain tissues were harvested and made into slices,stained with immunohistochemical techniques(SP)and photographed under the light microscope.The images were analyzed with the image analytical system and the data were statistically analyzed.Results:In the control group,positive staining of SYN was found in the entorhinal area,subiculum,stratum plextiforme,polymorphic layer of gyrus dentatus,stratum oriens,and stratum radiatum of hippocampus;weak positive staining of SYN was noticed in the stratum lacunosum-moleculare of hippocampus;positive staining of SYN was also found the membrane of pyramidal cells and granule cells,with the mean gray scale value of the hippocampal structure being 132.84?8.67.Positively stained neurons was also found in the entorhinal area and the subiculum,with a intensity of(7.80?1.03)/ mm~2.In the experiment group,the suhiculum and polymorphic layers of gyrus dentatus were positively stained for SYN;the entorhinal area,stratum oriens,stratum radiatum and stratum lacunosum-moleculare of hippocampus were strongly positive of SYN;the membrane of pyramidal cells and granule ceils were also strongly positive of SYN,with the mean gray scale value of the hippocampal structure being 116.27?5.70.Strongly stained neurons were also found in the entorhinal area and the subiculum,with the intensity being(11.90?1.45)/mm~2.The number of SYN positive neurons and the intensity of SYN in the experimental group were higher than those in the control group(P