Objective To investigate the relationship of lymphatic vessel density (LVD)and lymphatic metastasis in gastroduodenal neuroendocrine carcinoma.Methods The intratumor lymphatic vessel deusity(iLVD) and peritumor lymphatic vessel density(pLVD) of 55 surgical gastroduodenal neuroendocrine carcinoma tissue samples were detected by immunohistochemistry(with monoclona1 antibody D2-40).The relationship of lymphatic metastasis and pLVD,iLVD was analyzed.Results The value of pLVD and iLVD was 31.02 ± 1.22 vs 5.29 ± 0.76 for gastroduodenal neuroendocrine carcinoma with lymph node metastasis,while it was 20.43 ± 1.39 vs 5.64 ± 1.01 for gastroduodenal neuroendocrine carcinoma without lymph node metastasis,showing pLVD was significantly higher than iLVD in both groups.pLVD of gastroduodenal neuroendocrine carcinoma was closely related to lymph node metastasis (P =0.001,r =0.872),the differentiation of carcinoma,and lymphatic involvement while iLVD of gastroduodenal neuroendocrine carcinoma had no relation with lymph node metastasis(P =0.293).Conclusion Compared to iLVD detection,pLVD detection is more valuable in gastroduodenal neuroendocrine carcinoma since it can help to predict lymph node metastasis status and the prognosis.

Objective To describe the clinicopathologic features of two cases of mucoepidermoid carcinoma of the skin.Methods Two cases of mucoepidermoid carcinoma of the skin were analyzed histopathologically using hematoxylin and eosin (HE) staining,alcian blue-periodic acid Schiff (AB-PAS) staining and immunohistochemical staining.Relavant literature was reviewed.Results Histopathological examination showed that the tumor was subcutaneously located in both cases,with epidermoid cells and intermediate cells arranged in sheets or nests,as well as different sizes of glandular structures lined by mucinous columnar epithelium in some areas.Both tumors had a relatively clear boundary with peripheral invasive growth and no obvious capsules.Immunohistochemical examination showed positive staining for carcinoembryonic antigen (CEA),high and low molecular weight cytokeratin (CK(H) and CK(L)).The cytoplasm of mucous cells was stained blue with,and mucus was visualized after,AB-PAS staining.Conclusions Primary mucoepidermoid carcinoma of the skin is a kind of malignant tumor arising from skin appendages,whose diagnosis depends on histological and immunohistochemical examination.

Objective To develop an immunohistochemical assay for the diagnosis of cutaneous malignant melanoma (CMM) micrometastasis via blood and lymphatic vessels,and to evaluate its clinical significance.Methods Fifty-three patients (32 males and 21 females) histopathologically diagnosed as CMM were enrolled in this study.The patients were aged (61.2 ± 8.4) years (range,52-72 years).Tissue specimens were obtained from the central area of tumor in each case,and also from removed lymph nodes in some cases.The average duration of follow-up was (65.00 ± 5.68) months.During the follow-up,17 patients died of the recurrence or metastasis of CMM,and 6 patients were lost to follow-up.The expressions of D2-40,S100 and CD34 antigens in 53 tissue specimens were examined by immunohistochemical staining with three individual monoclonal antibodies,or by an immunohistochemical method using 2 two-antibody cocktails (D2-40/S 100 and CD34/S100) and double-color chromogens in single tissue sections.Results Of the 53 patients,30.19％ (16/53) were positive for hematoxylin-eosin (HE) staining combined with immunohistochemical staining with individual monoclonal antibodies,and 49.06％ (26/53) for the immunohistochemical method using two-antibody cocktails and double-color chromogens.Statistical differences were found in the positive rate between the two methods (x2 =3.94,P＜ 0.05).Compared with patients without blood/lymph vessel tumor emboli,those with blood/lymph vessel tumor emboli showed higher lymph node metastasis rate (80.77％ (21/26) vs.37.04％ (10/27),x2 =10.43,P ＜ 0.001),but lower five-year survival rate (42.31％ (11/26) vs.70.37％ (19/27),x2 =4.25,P ＜ 0.05).Conclusions The immunohistochemical method with two-antibody cocktails is superior to HE staining combined with immunohistochemical staining with individual monoclonal antibodies in the detection of blood/lymph vessel tumor emboli.And blood/lymph vessel tumor emboli may be an important prognostic factor in patients with CMM.

Objective To study the pathologic features,differential diagnoses and pathogenesis of splenic vascular tumors.Methods The records of 7 patients with splenic vascular tumors were reviewed.Immunohistochemical studies were performed by EnVision assay.Results The histopathologic diagnoses were hemangioma (n=2),lymphangioma (n=2) and hemolymplangioma (n=3 ).Positive immunoreactivity of CD31 and CD34 was found in vascular endothelial cells,and D2-40 expression was only identified in lymphatic endothelial cells.Conclusions Immunohistochemical staining of D2-40,CD31,CD34 was very helpful in the diagnosis of splenic vascular tumors.

A 69-year-old male patient presented with a gradually enlarging mass in the left inner upper thigh for more than 2 months,and pigmented patches in the left medial leg for more than 20 years.Physical examination revealed a painless mass measuring 3 cm × 2 cm × 2 cm in size in the left inner upper thigh.Several pigmented patches were observed in the left medial leg,and the largest pigmented patch measured 2 cm× 2 cm in size with an irregular border and uneven pigmentation.The mass in the left inner upper thigh was resected and subjected to histopathological examination,which showed proliferative epithelioid neoplastic cells with mucous matrix,round and spindle cells of varying sizes separated by mucous matrix.The immunohistochemical study of tumor cells showed positive staining for vimentin,S100 and Melan-A,but negative staining for actin,desmin,CD56,epithelial membrane antigen,cytokeratin,leukocyte common antigen,CD99,chromogranin A and synaptophysin.Hematoxylin-eosin staining of pigmented patches on the left medial leg revealed squamous epithelium covering the surface of lesions with no superficial ulceration or atypia in epithelial cells,unevenly distributed melanophages,fibroplasia accompanied by collagen formation,obviously decreased skin appendages,infiltration of a few inflammatory cells in the dermis.AB-PAS staining was negative.The immunohistochemical study of pigmented patches showed positive staining for vimentin and Melan-A.The patient was pathologically diagnosed with metastatic myxoid melanoma with partial regression of the primary lesion.

Objective To investigate the dormancy reason of the seeds by studying on intrinsic inhibitor in the seed of Paeonia lactiflora.Methods Biological methods and embryo culture were carried out to study the activity of intrinsic inhibitor in the testa and endosperm of the P.lactiflora seeds.ResultsThe Activity of intrinsic inhibitor exists in the testa and endosperm,and both of the intrinsic inhibitors showed stronger inhibitory effect on the growth of younger roots of Brassica pekinensis than that on their seed germination.The Rf 0.5 section of the ether extract of endosperm showed the strongest inhibition on the growth of younger roots of B.pekinensis(relative rate: 36.36%).All embryos that inoculated on different culture medium could root,but the epicotyls of embryos that inoculated on culture medium without GA3 could not elongate,which showed that the embryo in vitro also exists the dormancy of epicotyl,or embryo with inhibitor.Conclusion Intrinsic inhibitor in the seed of P.lactiflora is the main factor that results in its dormancy.

Objective To explore the relationship between pituitary tumor transforming gene(PTIG) and Ki67 expression and their significance in multiple adolescent breast fibroadenomas (fibroadenomatosis)and single persons(fibroadenomas) and their clinical value.Methods A total of 53 fibroadenomatosis and 160 fibroadenomas specimens in our hospital were collected.All the samples were fixed with 10％ formalin and then embedded with paraffin.The protein expression of PTTG and Ki67 was detected by immunohistochemical staining.The relation between expression of PTTG and Ki67 in fibroadenomatosis was analyzed.Results The positive rate of PTTG and Ki67 was higher in fibroadenomatosis than in fibroadenomas (P ＜ 0.005).Conclusions High expression of PTTG and Ki67 in fibroadenomatosis leads to the proliferation of tumor cell and upregulates tumor angiogenesis.High expression of PTTG and Ki67 may play important roles in tumorigenesis and pathological process of fibroadenomatosis.

Objective To explore the method of primary myometrial cell culture to provide a valuable cell model for a series of research works. Methods Human mometrial cells were isolated by cutting and collagenase digestion, and incubated with DMEM medium and fetal bovine serum. The first passage cells were identified by anti-desmin and-?-action antibodies immunohistochemical staining. Results Primary cultivated human myometrial cells had high purity and viability. Conclusions Primary culture human myometrial cells could obtained by cutting and collagenase digestin.

Objective:To investigate the influence of Mannan Peptide combined with 5-Fu on the proliferation and apoptosis of HepG2 cells and to explore the related mechanism.Methods:HepG2 cells were divided into four groups:the control group,5-Fu group,Mannan Peptide group and combination group.Cell proliferation was evaluated by MTT assay.Flow cytometry was used to examine the cell cycle,proliferation index(PI)and apoptosis ratio.The expression of Bcl-2 protein in each group was analyzed by immunocytochemical method.Results:The inhibitive effect of 5-Fu plus Mannan Peptide was significantly stronger than that of Mannan Peptide alone,in a time-and dose-dependent manner. Cells were treated with medicine for 48 hours.Compared with cells in the control group and 5-Fu group,the percentage of cells in G_0/G_1 stage and the apoptosis ratio in the combination group were increased(P<0.01).In immunocytochemical analysis,the combination of Mannan Peptide and 5-Fu could significantly decrease Bcl-2 protein level(P<0.01).Conclusion:The combination of Mannan Peptide and 5-Fu can synergistically suppress the proliferation and induce apoptosis of human hepatoma HepG2 cells.The mechanism might be associated with down-regulation of Bcl-2 protein expression.