Objective To explore the effect of fibroblast growth factor receptors 1-dominant negative strategy (FGFR1-DN) on alkaline phosphatase (ALP) activity of bone marrow stromal stem cells (BMSCs) after osteogenic induction.Methods BMSCs were transfected with eukaryotic expression plasmid pcDNA 3.1 (+)-DN FGFR1 and pcDNA3.1 (+)-FGFR1.The experiment was conducted in 4 groups:FGFR1-DN transfection group,FGFR1 transfection group,pcDNA3.1(+) empty vector transfection group and non-transfection group.The ALP activity of BMSCs was detected in logarithmic growth phase after osteogenic culture.The qualitative detection of ALP activity was carried out immunohistochemically while the quantitative detection by cALP kit.The ALP activity was compared between the 4 groups at 7 and 14 days after osteogenic induction.Results Compared with 7 days,the ALP activity at 14 days was significantly increased in the 4 groups,and the increase in FGFR1-DN transfection group was significantly higher than in the other 3 groups (P ＜ 0.05).At both 7 and 14 days,the ALP activity in FGFR1-DN transfection group was the highest while that in FGFR1 transfection group was the lowest (P ＜ 0.05).Conclusions FGFR1-DN can promote the ALP activity of BMSCs during osteogenesis.This may provide an experimental basis for the joint application of local gene therapy and tissue engineering and for construction of tissue engineered bone with better biocompatibility.

Purpose To observe the histopathologic changes of acquired immure deficiency syndrome (AIDS) in a Chinese Rhesus monkeys model and the pathogenesis that initiated the changes.Methods Chinese Rhesus monkeys were sacrificed after being inoculated SIVmac239 by Ⅳ(n=2)for four months.Autopsy was carried out by pathologic routine method.The lymph nodes, heart, liver, spleen, lung, kidney, digestive tract and other tissues were selected, the tissues fixed with 10% neutral formalin, and the pathologic sections were prepared by HE staining and immunohistochemical staining and special staining after paraffin imbedding.Results The main histopathological changes appeared in the immune system in different organs. The lymph nodes began to display the complex changes in a short period of time infected by the virus, including proliferation of lymphoid follicles, atrophy, or both; some lymphoid follicles of lymph nodes had few lymphocytes, with fibrous hyperplasia and immune complex (IC) deposition, displaying a burning down phenomenon.Splenomegaly and blood vessel and its endothelial cell proliferation in splenic corpuscles were noted with the immune complex deposition. Other parts of the mucosa-associated lymphoid tissue had different degrees of hyperplasia, or atrophy.Conclusion Histopathologic changes in Chinese Rhesus monkeys infected by SIVmac239 strain are very similar to human AIDS, which suggests that the model is a useful tool for the prevention and treatment study of AIDS.

Purpose To study the relationship between Epstein-Barr virus ( EBV) and breast cancer. Methods 246 cases of breast lesions at different development stages were selected and EBV DNA, RNA and protein was used by polymerase chain reaction ( PCR) , in situ hybridization ( ISH) , laser capture microdissection ( LCM) , immunohistochemistry ( IHC) EnVision technology. Results No expression of EBV latent membrane protein LMP1 was detected in all 246 cases of benign and malignant breast lesions. In 12 cases of breast cancer of EBV DNA, carcinoma in situ and breast lesions not EBV DNA was detected by PCR. However, using digoxigenin la-beled EBV DNA probe for the 48 cases ( including 12 cases of breast cancer specimens of positive PCR amplification) of benign and malignant breast lesions, no positive hybridization signal was detected in cancer cells, mammary epithelial cells and stromal lympho-cytes. Using laser capture microdissection and PCR amplification, cancer cells and stromal cells were captured respectively from 12 ca-ses of PCR positive and 12 cases of PCR negative of breast cancer specimens, we found EBV DNA was only amplified in mesenchymal cells. In the detection of the EBER expression with EBV RNA probe and in situ hybridization, the results of 75 cases of benign and malignant breast lesions ( including 12 cases of breast cancer by positive PCR amplification) were all negative. Conclusions The re-sults indicate that the tumorigenesis and development of breast cancer have nothing to do with EBV infection in all cases were chosen.

Objective To investigate the mineralization impact of fibroblast growth factor receptors 1 dominant negative (FGFR1DN) on osteogenic induction culture of bone marrow stromal cells (BMSCs).Methods The 3rd generation BMSCs were divided into 4 equal groups (n =24).FGFR1-DN group was transfected by pcDNA3.1 (+)-FGFR1DN,FGFRI group by pcDNA3.1 (+)-FRFR1,blank load group by pcDNA3.1 (+)-blank vehicle and non-transtection group by nothing.After successful transfection was confirmed when the cells were in the logarithmic phase,osteogenic induction culture was conducted continuously for 21 days.Mineralized nodule formation was observed by alizarin red staining.The amount of mineralized material was calculated according to the standard curve of alizarin red concentration.Results Continuous osteogenic induction for 21 days showed on the bottom of the hole visible round opaque calcified nodules after alizarin red staining.The BMSCs in the FGFR1-DN group induced in the logarithmic growth phase by osteoblasts exhibited significantly increased osteogenic capacity while those in the FGFR1 group displayed diminished osteogenic capacity.The concentration of alizarin red was the highest in the FGFRI-DN group (1.33 ±0.19),the lowest in the FGFRI group (1.00 ± 1.17),and moderate in the blank load group (1.20 ± 0.16) and non-transfection group (1.17 ± 0.17),showing significant between-group differences (P ＜ O.05).Conclusions FGFR1-DN can promote cell proliferation in the early differentiation of BMSCs and also mineralization of osteoblasts in bone induction culture after the logarithmic growth phase.This may provide a hint for local gene treatment of bone defects.

Neuroendocrine carcinoma of the breast is a rare malignant tumor which, with the features of Merkel cells is even rarer. Herein, we report a case of small cell carcinoma with Merkel cell features in a 52-year-old female. Microscopically, the tumor was characterized by diffuse and consistent small round cells that were de-adherent. The tumor cells had round or oval nuclei with delicate chromatin and small nucleoli, the cytoplasm was sparse and eosinophilic. Additionally, the tumor was accompanied by high-grade ductal carcinoma in situ. Immunohistochemical staining showed that infiltrating tumor cells were positive for neuroendocrine markers, and punctately positive for CK20. The patient underwent modified radical mastectomy, axillary lymph node dissection, and postoperative adjuvant chemotherapy. No recurrence or metastasis was observed during follow-up period. Primary breast small cell carcinoma with Merkel cell features is rare and easily misdiagnosed as Merkel cell carcinoma. Early diagnosis and treatment may improve patient prognosis.

Objective:To investigate the intra-observer reproducibility of Ki-67　　　assessment in breast cancers using three methods based on digital slide.Methods:Thirty cases of invasive breast cancer tissues were immunostained for Ki-67 by automatic stainer, and then scanned into digital pathological slides. Ki-67 positive index was measured individually by three pathologists using size-set visual assessment of hot spot (SSVAHS), size-set semi-automatic counting of hot spot(SSSACHS), and size-set automatic counting of hot spot (SSACHS), respectively, and repeated for 10 times. Intraclass correlation coefficient (ICC) of each assessment method was calculated, and the intraobserver reliability was classified as excellent, good, fair and poor according to ICC.Results:The ICC by 3 pathologists using SSVAHS was 0.832, 0.843 and 0.826, respectively, The ICC using SSSACHS was 0.926,0.938,0.929, and the ICC using SSACHS was 0.964, 0.971 and 0.968.The intraobserver reliability level of all three methods was excellent.Conclusion:The three methods of Ki-67 assessment achieve satisfactory intraobserver reproducibility, and the order of reproducibility from high to low is SSACHS, SSSACHS, and SSVAHS.

Objective To investigate the mutation status of epidermal growth factor receptor ( EGFR) and KRAS gene in patients with non-small cell lung cancer ( NSCLC) in Xuanwei, Yunnan and to correlate the mutation status with clinicopathologic features.Methods Mutation status of exons 18, 19, 20 and 21 of EGFR, and codons 12, 13 of KRAS in 63 cases of NSCLC were analyzed by gene sequencing and ARMS-Taqman probe method.Correlation with patients′clinicopathological characteristics was performed. Results EGFR and KRAS mutations were present in 55.6% (35/63) and 6.3% (4/63), respectively. EGFR gene mutations were present, including exon 18 G719X in 14.3% (5/35), exon 19 in 14.3%(5/35), exon 20 S768I and T790M in 20.0% (7/35), exon 21 L858R in 31.4% (11/35), exon 18 G719X and exon 20 S768I double mutation in 17.1%(6/35), and exon 20 T790M and exon 21 L858R double mutation in 2.9%(1/35).KRAS mutations were seen in codon 12 in 3 of 4 cases, and codon 13 in 1 of 4 cases.EGFR mutations were mutually exclusive with KRAS mutations.According to statistic analysis, EGFR mutations were associated with the histological types of NSCLC ( P<0.05 ) , but without correlation with patient′s gender, age, smoking status and lymph node metastasis ( P >0.05 ) .KRAS mutations in NSCLC had no correlation with the clinical pathologic characteristics of the patients.Conclusions A higher frequency of EGFR exon 18 G719X and 20 exon S768I mutations are found in the patients in Xuanwei, Yunnan.EGFR mutations are associated with histologic types of NSCLC, but without correlation with patient′s gender, age, smoking status and lymph node metastasis.KRAS mutation in NSCLC has no correlation with the clinicopathologic characteristics of the patients.