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Objective:To investigate the role of Caspase-1/gasdermin D (GSDMD) -mediated cell pyroptosis in anti-tumor effect of cisplatin (DDP) in triple-negative breast cancer (TNBC) .Methods:HE staining and immunohistochemical staining were performed to detect the morphological changes and the expression of pyroptosis/apoptosis pathway related proteins in TNBC tissues before and after DDP-based neoadjuvant chemotherapy (NACT) . The TNBC cell line MDA-MB-231 was treated with DDP and the morphological changes were observed. The type of cell death induced by DDP was analyzed by Annexin V-FITC/PI double staining and flow cytometry. Lactate dehydrogenase (LDH) release assay and ELISA were performed to detect the release of LDH and inflammatory factors (IL-18 and IL-1β) in cell culture supernatant after DDP treatment. Western blot (WB) was performed to detect the expression of pyroptosis/apoptosis pathway related proteins in cells after DDP treatment. MDA-MB-231 cells treated with DDP were co-treated with caspase-1 specific inhibitor to inhibit pyroptois or co-treated with caspase-3 specific inhibitor to inhibit apoptosis. The effect of caspase-1 inhibitor or caspase-3 inhibitor on the anti-tumor effect of DDP was detected by MTT assay, clone formation assay, transwell assay and would healing test.Results:Reactive changes in the breast surgical specimen after DDP-based NACT included cell swelling and inflammatory cell aggregation around the tumor bed, which were more similar to pyroptosis. The up-regulation of key molecules of pyroptosis pathway post-NACT was significantly higher than that of key molecules of apoptosis pathway. Further experiments in vitro showed that DDP could induce MDA-MB-231 cells to show pyroptosis-like changes characterized by large bubbles blowing from the cellular membrane. Flow-cytometry analyses showed that the death type of MDA-MB-231 cells caused by DDP was mainly Annexin V +PI + cells (mainly lytic cells, such as pyroptosis) . Additionally, DDP treatment induced significant activation of caspase-1 and GSDMD, increased the release of LDH, IL-18 and IL-1β, however, the activation level of caspase-3, which dominates the apoptosis pathway, was significantly lower than that of caspase-1/GSDMD. Moreover, caspase-1 inhibitors (blocking the classical pyroptosis pathway) had a significantly greater inhibitory effect on the anti-tumor effect of DDP than caspase-3 inhibitors (blocking the apoptosis pathway) . Conclusion:Caspase-1/GSDMD mediated pyroptosis may play a leading role in the anti-tumor effect of DDP in triple-negative breast cancer.
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Objective:To investigate the diagnostic value of GATA binding protein 3 (GATA3) , mammaglobin (MGB) and gross cystic disease fluid protein-15 (GCDFP-15) in breast cancer.Methods:Immunohistochemistry was performed to detect the expression of GATA3, MGB, GCDFP-15, estrogen receptor (ER) , progesterone receptor (PR) , human epidermal growth factor receptor 2 (HER2) and Ki-67 in breast cancer. The breast cancer was divided into primary/metastatic breast cancer, hormone receptor positive/negative, or different molecular types of breast cancer according to the tissue origin and immunophenotype. The positive rates of GATA3, MGB and GCDFP-15 were statistically analyzed.Results:The positive rate of GATA3 in primary/metastatic breast cancer (92.5%; 94.25%) was significantly higher than that of MGB (42.11%; 29.17%) and GCDFP-15 (55.77%; 31.34%) ( P<0.01) , and GATA3 had better sensitivity for evaluating the tumor of unknown origin, especially in malignant hydrops. In addition, the positive rate of MGB and GCDFP-15 in hormone receptor negative and triple negative breast cancer was low, which further increased the difficulty of diagnosis of these two kinds of metastatic breast cancer. However, the positive rate of GATA3 in hormone receptor negative (90.38%) or triple negative breast cancer (85.29%) was significantly higher than that of MGB (20.00%; 5.00%) and GCDFP-15 (35.09%; 33.33%) ( P<0.01) , suggesting that it had great potential diagnostic value in hormone receptor negative and triple negative metastatic breast cancer and was a specific marker superior to MGB and GCDFP-15. Conclusion:GATA3 has a higher diagnostic value than MGB and GCDFP-15 for the assessment of tumors of unknown origin, especially in malignant hydrops, or breast cancer with an immunophenotype of hormone receptor negative and triple negative breast cancer.
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Objective To investigate the expression and significance of CD68,E-cadherin and transforming growth factor-β1 (TGF-β1) in breast cancer.Methods Immunohistochemistry was used to detect the expression of CD68,E-cadherin and TGF-β1 in breast cancer tissues.The relationship between the expression of proteins with the clinicopathological parameters and the recurrence of breast cancer was analyzed.And the correlation among the three proteins was also analyzed.Results The positive expression rate of CD68 and E-cadherin was 70.4％ and 72.6％,respectively,which was significantly higher than that in adjacent tissues(CD68:x2=44.278,P=0.000;TGF-β1:x2=121.529,P=0.000).The positive expression rate of E-cadherin in breast cancer was 29.1％,which was significantly lower than that in adjacent tissues(x2=244.965,P=0.000).The expression of CD68 protein was closely related to clinical stage (x2=11.720,P=0.003),lymph node metastasis (x2=9.394,P=0.002) and ER status (x2=5.204,P=0.023).The expression of E-cadherin protein was significantly correlated with the histological grade(x2=6.561,P=0.038) and lymph node metastasis(x2=6.892,P=0.009),and the expression of TGF-β1 protein was significantly correlated with the histological grade(x2=6.427,P=0.040) and ER status(x2=5.755,P=0.016) of patients.The three proteins were all significantly related to the 5 year recurrence of breast cancer (CD68:γ=0.152,P=0.021;TGF-β1:γ=0.157,P=0.017:E-cadherin:γ=-0.145,P=0.028).The expression of CD68 protein was negatively correlated with E-cadherin protein (γ=-0.151,P=0.022),and positively correlated with the expression of TGF-[β1 (γ=0.200,P=0.002).E-cadherin protein was negatively correlated with the expression of TGF-β1 protein(γ=-0.143,P=0.031).Conclusion CD68 positive cells (tumor associated macrophages,TAMs),E-cadherin and TGF-β1 may be involved in the progression of breast cancer,detection of the expression will provide the theoretical basis and guidance for the clinical pathological diagnosis and treatment of breast cancer.
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Objective To compare the differences and correlation between immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH) of HER2 in invasive breast cancer.Methods A total of 604 breast cancer specimens from Nov.2014 to Dec.2016 in Renmin Hospital of Wuhan University were evaluated for HER2 status by both IHC and FISH to calculate the concordance between these two assays and analyze the causes of discordant cases.Results Except for cases with IHC uncertainty(2+),overall concordance between IHC and FISH in detection of HER2 was 97.2％ (412/424).The concordance for positive and negative detection was 93.0％(146/157) and 99.6％(266/267),respectively.The main reasons for the difference of HER2 expression in breast cancer detected by the two methods were the incorrect interpretation of IHC,the higher or lower concentration of antibody,and the change in guidelines.Conclusion HER2 IHC assay is subjective and sensitive to technical factors for determining the expression levels of HER2 protein in breast cancer so that it is not as stable as HER2 FISH assay.
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Objective To explore the application value of telepathological consultation in helping grassroots hospitals.Methods 578 cases of intraoperative telepathology consultation were reviewed,and the accuracy and the timely rate of diagnosis were calculated.The systematic distribution,benign and malignant distribution,and the distribution difference in different primary hospitals were analyzed,so as to evaluate the popularization value of the intraoperative telepathology consultation.Results The accuracy rate of 578 cases of intraoperative telepathology consultation was 99.83％.The timely rate of consultation in 30min was 96.02％,and most reports could be diagnosed in 2 to 5 mins.The source of tissues involved in consultation were thyroid,breast,ovary/fallopian tube and lung.In all cases,24.39％ of the malignant tumors were found.Among the diseases of different systems,the proportion of malignant tumors is the highest in breast diseases,followed by lung,thyroid and ovary.Among the four hospitals with most of the consultations,the rate of malignant tumor in Renmin Hospital of Jianli County was the highest,followed by Renmin Hospital of Yingshan County,Renmin Hospital of Xiaochang County,and Fifth Division Hospital of Xinjiang.Conclusion Intraoperative telepathology consultation can provide accurate and timely expert consultation for grassmots hospitals,avoid the "second operations" of the patients,improve the access of medical treatment for people living in relatively remote areas,solve the shortage of pathologists at the grassroots hospitals,and improve the level of doctors' diagnosis and treatment at the grassroots hospitals,which is worth popularizing and applying in Pathology Department of the grassroots hospitals.
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Objective To detect the expression profile of transcription factor C/EBPβ in human immortalized normal hepatic cell lines and hepatocellular carcinoma cell lines so as to determine the correlation between C/EBP3 with cell death mediated by endoplasmic reticulum stress in hepatocellular cells.Methods We cultured the human immortalized normal hepatic cells lines HHL5 and HL7702 and hepatocellular carcinoma cell lines SMMC7721;Bel7402,HepG2 and Hep3B.Hep3B cells were used as the cell model in tunicamycin-induced endoplasmic reticulum stress.Cellular morphology was observed under an inverted optical microscope.MTT assay was used to assess the inhibition of cell growth.To detect cell apoptosis,the cells were dyed with Hoechst 33258 and observed using a fluorescence microscope.RToPCR and Western blotting were used to detect the expression of at mRNA and protein levels,respectively.Results We found that normally the mRNA and protein isoform of C/EBPβ,C/EBPβ-1,were both expressed in all of the four hepatocellular cell lines and the two immortalized normal hepatic cell lines,while C/EBPβ protein isoform C/EBPβ-3 was only expressed in the two immortalized normal hepatic cell lines.Tunicamycin increased the expressions of both mRNA and protein of C/EBPβ in Hep3B cells and the increase of protein isoform C/EBPβ-3 was the most remarkable.In Hep3B cells,cell death was induced by tunicamycin through endoplasmic reticulum stress activity.Apoptosis as well as paraptosis was observed in tunicamycin-induced cell death.Conclusion C/EBPβ-3,one of the protein isoforms of C/EBPβ,is only expressed in normal hepatic cell lines,but not in hepatocellular cell lines.C/EBPβ is involved in cell death mediated by endoplasmic reticulum stress activity in hepatocellular carcinoma cells.
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Purpose To explore the clinicopathological features,immunophenotype,differential diagnosis and prognosis of the composite pheochromocytoma (CP)-ganglioneuroma.Methods 3 cases of CP-ganglioneuroma were stained by immunohistochemical SP method,and the related literatures were reviewed.Results 3 cases of CP-ganglioneuroma were one male and 2 females,the age were 37-64.Case 3 were of primary mediastinal.Microscopically,the tumor tissues were composed of two components:one type of tumor cells were arranged in nests with a predominant Zellballen pattern,round or oval nuclei,fine granular cytoplasm and rare mitotic,another part of the neoplasm showed scattered and aggregated distributed ganglion cells in the background of neurofibromatosis which aligned bundles and interwoven,the edge of the tumor was still residual adrenal tissue.Immunohistochemically,components of pheochromocytoma were positive for CD56,CgA,Syn,vimentin and negative for SMA,Melan-A,α-inhibin NF with low Ki-67 proliferation index.S-100 was positive in supporting cells,ganglioneuroma components were positive for NF,S-100 with low Ki-67 proliferation index.CgA and Syn were weakly positive or negative in the ganglion cells.Conclusion CP is a relatively rare tumor,which can not be distinguished from pheochromocytoma in clinical and radiological diagnosis.The corresponding clinical treatment and follow-up management should be taken according to the different ingredients (benign or malignant).
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Objective To compare the clinical evaluation in diagnosis of thyroid nodules between fine needle aspiration(FNA) and core needle biopsy(CNB).Methods 82 cases with thyroid nodules undergoing FNA and 33 cases with thyroid nodules undergoing CNB were selected.Cytology,histopathology,and postoperative results were compared with postoperative pathology as the gold standard.Results The accuracy,sensitivity,specificity,positive predictive value,negative predictive value,false positive rate,false negative rate of FNA and CNB were 91.5％ vs 87.9％,93.2％ vs 87.5％,87.0％ vs 88.9％,94.8％ vs 95.5％,83.3％ vs 72.7％,13.0％ vs 11.1％,6.8％ vs 12.5％,respectively.There was no significant difference between FNA and CNB (P＞0.05).Conclusion Both of FNA and CNB examination show high accuracy,sensitivity and specificity in diagnosis of benign and malignant thyroid nodules,but FNA is more economical,safe,convenient and thus has more extensive clinical application.For lymphoma or borderline tumor,CNB examination combined with immunohistochemistry is of more advantages.
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Objective To observe the expression of transforming growth factor factor-β1 (TGF-β1) and vas cular endothelial growth factor (VEGF) in breast cancer to explore the correlation between each other and their relationship with invasion,metastasis and prognosis of breast cancer.Methods Immunohistochemistry was used to detect the expression of TGF-β1 and VEGF in 230 breast cancer tissues and adjacent tissues.The correlation between them and the relationship between their expression and the clinical pathological parameters of breast cancer were analyzed.Results The positive rates of TGF-β1 and VEGF in breast cancer were 72.6％ and 68.3％,respectively.But in adjacent breast tissues,the positive rate was 21.3％ and 42.2％,respectively.The expression of TGF-β1 was closely related to the histological grade,ER state and 5-year recurrence (P＜0.05),while ER status and 5-year recurrence were independent predictors of the positive expression of TGF-β1.The expression of VEGF was significantly correlated with ER status and 5-year recurrence (P＜0.05),while ER status was independent predictor of the positive expression of TGF-β1.The expression of VEGF was significantly positively correlated with the expression of TGF-β1 (γ=0.419,P＜0.01).Conclusion The expression of TGF-β1 and VEGF in breast cancer is closely related to the invasion,metastasis and prognosis of breast cancer,which has a certain clinical significance.
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Objective To explore the clinicopathological features and diagnosis of adenoid cystic carci noma of the breast (ACCB).Methods Clinical data of 3 cases of ACCB were collected and followed up to observe the clinical pathological features.Immunohistochemistry was used to determine the immune phenotype.Results All the 3 cases were female,aging from 55 to 72 years old (61 years old as the average).The tumors were found in subareolar region well-circumscribed.ACCB was characterized by the presence of a dual cell population of luminal and basaloid cells arranged in cribriform and tubular-trabecular patterns.On immunohistochemical staining,the myoepithelial cells expressed SMA,CK5/6,P63,CD117 and the glandular epithelium cells were mostly positive for CK18,EMA,CK7,while no expression of ER,PR,HER-2,CD10 was observed.Conclusions ACCB is a rare neoplasm with a triple-negative,basal-like phenotype,but exhibits an indolent clinical behavior.The diagnosis can be worked out by evaluation of clinical characterstics,histological and immunohistochemical features.
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Objective To investigate the diagnostic value of the instant gastrointestinal ultrasound contrast agent in the digestive diseases. Methods Five hundred and seventy-nine patients received the examination of the color Doppler after they drinked the ultrasonic contrast agent. Then the results were analyzed by consistency analysis. Results There was high consistency between the two examinations in the normal control, gastritis, peptic ulcer, gastric cancer and gastric leiomyoma. The Kappa value was 0.768, 0.913, 0.925, 0.939 and 1.000, respectively. But the consistency in the gastric polyp was low , the Kappa value was 0.368. Conclusion The color Doppler through the instant gastrointestinal ultrasound contrast agent has high diagnostic value in the common diseases of the digestive system.