Objective To investigate the clinical and pathological features of anorectal malignant melanoma ( AMM) .Methods The clinical pathological and follow-up data of 7 cases of anorectal malignant melanoma were collected.The clinical and histopathological features and immunophenotype were analyzed .Results Among the 7 cases, 5 cases were female, 2 cases were male, aged 50-68 years, the average was 61 years.The tumors located in the lower rectum in 1 cases, 2 cases of teeth near the line , 4 cases of anal anus .The structure and morphology of the cancer cells were diverse .The organizational structures were mainly diffuse , focal and acinar .The cellular morphologies were mainly epithelioid , spindle and lymphocyte like .And the results of IHC showed the expression of the following three markers, vimentin, S-100 and HMB45, were strongly positive in cancer cells.However, the expression of CK , EMA and LCA were negative .And 2 cases of CEA expression were positive , LI Ki-67 was 20%-50%.Conclusions Anorectal malignant melanoma is one kind of rare malignant tumor and its characteristic morphology , immunophenotype could be helpful for diagnosis and differential diagnosis .

Objective To evaluate the value of DNA quantitative analysis in diagnosis of breast tumor.Methods The data of 128 cases diagnosed with breast disease from Feb.2005 to Jan.2006 in Department of Pathology in Wuhan Center Hospital were collected,among whom 72 cases had benign breast lesion,10 cases had boundary breast lesion and 46 cases had malignant breast disease.Automated DNA imaging cytometry was used to identify DNA indexes of the 128 cases.Results The positive rate was 1.39％ (1/72),30.00％ (3/10) and 78.26％ (36/46) respectively in benign breast lesion,boundary breast lesion and malignant breast disease.The difference had statistical significance (P ＜ 0.05).In invasive ductal breast cancer,the positive rate was 57.14％ (4/7),76.47％ (13/17) and 100％ (18/18) respectively for breast cancer of grade Ⅰ,Ⅱ and Ⅲ.Conclusion Automated DNA imaging cytometry is of some value in analyzing the malignant grade of breast cancer and predicting the prognosis of breast diseases.

Objective To study the clinicopathological and immunohistochemical features of squamous cell carcinoma (SCC) of breast.Methods Three cases with breast SCC were studied by analyzing their clinical data,histomorphology and immunophenotype.The related literature was reviewed.Results The patients were all females.Their ages ranged from 37 to 50 years,with 43 years as the medium.The tumor diameter was 2-9 cm,with 4.3 cm as the medium.Metastasis of axillary lymph node was found in one case,accounting for 33％ of the total.Two cases had painless lumps in their left breasts.One case had a lump with local inflammation,pain and ulcer in her left breast.Histopathological examination showed the polygonal and spindle-shaped cells with eosinophilic cytoplasm were found in cancer nest,as well as the prominent intercellular bridges and keratin pearls were found to be rich in three cases.Interstitial fibrosis with hyalinization and lymphocytic infiltrate around the tumor was also found.Immunohistochemistry showed ER,PR,HER-2,CK18 and SMA were negative while CK,CK14,CK5/6,p63,S-100 and vimentin were positive.Conclusions SCC is a rare subtype of breast carcinoma with distinct histopathological and morphological features.SCC is a typical basal-like breast carcinoma,and the majority of breast SCC appear a basal-like immunophenotype with poor prognosis.

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.

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.

Objective To investigate the effects of hyperbaric oxygen (HBO) combined with neural stem cell (NSC) transplantation on functional recovery after spinal cord injury (SCI).Methods Forty healthy adult Sprague-Dawley rats were randomly divided into a sham group,an SCI group,an NSC group and an HBO+NSC group.In the latter three groups,complete spinal cord transection was performed at T10.The rats of the HBO+NSC group were treated with HBO therapy after NSC transplantation.Hindlimb motor function was assessed with the BassoBeattie-Bresnahan (BBB) scale and tilt table testing.Motor evoked potentials (MEPs) were measured to evaluate the rats'nerve conduction function.The amount of surviving transplanted NSCs at the injured site was counted using Hoechst dyeing.Results The average BBB scores and tilt table test results in the HBO+NSCs group were significantly better than in the SCI and NSC groups from the 2nd week post-SCI.By the 4th week the average latency period and MEP amplitude in the HBO+NSC group were significantly better than in the NSC group,while MEPs in the SCI group still had not re-appeared.The surviving amount of transplanted NSCs in the HBO+NSC group was also significantly greater than in the NSC group.Conclusion HBO combined with NSC transplantation has synergistic effects and can improve functional recovery after SCI.This may be explained by the important role of HBO in promoting the survival of transplanted NSCs.

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.

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.