Pilot study of PTEN deletion affecting the expression of Cu/Zn SOD / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate whether the deletion of PTEN affects the expression of Cu/Zn SOD and the related biology.</p><p><b>METHODS</b>Protein and mRNA expression levels of PTEN, P-Akt, Cu/Zn SOD in the control immortalized wild type mouse embryonic fibroblast cells (PTEN+/+) and PTEN-null cells (PTEN-/-) were evaluated by Western blot and Northern blot respectively. The level of superoxide anions were detected using fluorescent probes. The DNA damage was documented by single cell alkalescence gel assay. MTT was used to study the effect of H2O2 on the proliferation of cells.</p><p><b>RESULTS</b>The expression of Cu/Zn SOD was down regulated at both protein and mRNA levels, and the level of superoxide anions increased in the PTEN-null cells (PTEN-/-). The phosphorylation level of Akt kinase was up-regulated and the antiproliferative effect of H2O2 decreased in PTEN-/- cells. Furthermore, DNA damage was observed significantly severer in both the blank control and H2O2 treated groups than that in the PTEN+/+ cells.</p><p><b>CONCLUSIONS</b>Deletion of PTEN affects the expression of Cu/Zn SOD. As a result, reactive oxygen species (ROS) keep at a high level, along with decrease of accumulated oxidative damage and the antiproliferative effect of ROS.</p>

Artesunate suppresses human endometrial carcinoma RL95-2 cell proliferation by inducing cell apoptosis / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the inhibitory effect of artesunate on human endometrial carcinoma RL95-2 cell line proliferation in vitro and the possible mechanisms.</p><p><b>METHODS</b>The inhibitory effect of artesunate on the cell proliferation was assessed with MTT assay. Transmission electron miscrosopy was used to observe the morphological change of the cells after the treatment. Flow cytometry was performed to examine the changes in the cell cycle, reactive oxygen species (ROS) levels (with DCFH-DA labeling) and mitochondrial membrane potential (rhodamine123 staining), and caspase-3 activity was detected by immunohistochemistry.</p><p><b>RESULTS</b>Artesunate inhibited the proliferation of RL95-2 cells with an IC(50) of 26.29 microg/ml. Transmission electron microscopy revealed early apoptotic changes of the cells with obvious chromatin fragmentation. The cell cycle arrest at G(0)/G(1) phase was observed by flow cytometry, and immunohistochemistry demonstrated caspase-3 positivity in cytoplasm. ROS generation in the cells increased obviously after treatment with artesunate for 72 h, which also resulted in lowered mitochondrial membrane potential.</p><p><b>CONCLUSION</b>Artesunate suppressed the proliferation of RL95-2 cells in vitro possibly by inducing cell apoptosis.</p>

Expression of peroxisome proliferator-activated receptor gamma in glioma / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the expression and significance of peroxisome proliferators-activated receptor gamma (PPAR gamma) in human glioma.</p><p><b>METHODS</b>Immunohistochemical staining for PPAR gamma was performed using biopsy specimens of human glioma of various histological types. Expression of PPAR gamma and GFAP in glioma cell lines SWO-38, U251 and SHG-44 were analyzed using Western blotting and reverse transcription-polymerase chain reaction (RT-PCR).</p><p><b>RESULTS</b>Immunohistochemical study showed that PPAR gamma was expressed in glioma tissues with positive rate of 37.5%. Western blotting and RT-PCR showed that PPAR gamma was expressed in both glioma cell lines SWO-38 and U251, but not in SHG-44 cells. However, high expression of GFAP was detected in SHG-44 cells.</p><p><b>CONCLUSION</b>PPAR gamma is associated with carcinogens of glioma. Actived PPAR gamma by agonist may be a novel approach to the treatment of glioma.</p>

Primary diffuse large B-cell lymphoma of testis: a clinicopathologic study of 14 cases / 中华病理学杂志

<p><b>OBJECTIVE</b>To study the clinicopathologic features, immunohistochemical findings and prognosis of primary diffuse large B-cell lymphoma (DLBCL) of testis.</p><p><b>METHODS</b>Fourteen cases of primary DLBCL of testis, diagnosed according to the 2001 World Health Organization staging standards for hematopoietic and lymphoid tumors, were retrospectively studied. Immunohistochemical study was performed and follow-up information analyzed.</p><p><b>RESULTS</b>The median age of the patients was 62 years. There were 10 patients in stage I, 3 in stage II and 1 in stage IV. Follow-up information was available in 11 patients (78.6%). Three of which were still alive and eight died (with duration of survival ranging from 5 to 19 months). The patients usually presented with unilateral painless enlargement of testis. Histologically, the lymphoma cells of all cases showed a centroblastic appearance. One case belonged to the germinal center B cell-like subtype on immunohistochemical study, while the remaining 13 cases were classified as non-germinal center B cell-like subtype. Ten of the 14 cases (71.4%) showed overexpression of p53 protein. Most cases demonstrated high proliferation index. Six of the 14 cases (42.9%) were positive for bcl-2 protein. The overall 1-year, 2-year and 5-year survival rates were 45.5%, 17.0% and 17.0%, respectively.</p><p><b>CONCLUSIONS</b>Most cases with primary DLBCL of testis were of peripheral activated B-cell origin. The prognosis is usually not favorable, with propensity of local relapse and systemic dissemination. Accurate pathologic diagnosis relies on detailed histologic examination and immunohistochemical study.</p>

Heteroplasmy in human mtDNA control region / 法医学杂志

OBJECTIVE@#To observe the length heteroplasmy and point heteroplasmy in human mtDNA control region.@*METHODS@#The peripheral blood, buccal cell, and single hair shaft from 50 individuals and 16 family members, related in their maternallineage were analyzed by direct sequencing, and clones from 20 individuals whose mtDNA sequences have a T-C transition at 16189 nt were sequenced.@*RESULTS@#No point heteroplasmy were observed in peripheral blood, buccal cell, single hair shaft from the same individual, neither in maternally related individuals. Length heteroplasmy was observed in those individuals with a homopolymeric tract and the different clones from the same individual has different proportions of length variants, but the hair shafts from the same individual were very similar to the measurements made from blood DNA. No length heteroplasmy was observed between different tissues from the same individual.@*CONCLUSION@#mtDNA sequences have a characteristic of high consistency and genetic stability, mtDNA sequencing is a suitable tool for forensic applications such as individual identification.

The effects of genistein on epidermal growth factor receptor mediated signal transduction pathway in human ovarian carcinoma cells lines SKOV3 and its xenograft in nude mice / 中华病理学杂志

<p><b>OBJECTIVE</b>It was reported previously that genistein could inhibit proliferation of human ovarian carcinoma cell line SKOV(3), but mechanism was not clear. There is a close relationship between EGFR mediated signal transduction pathway and the development of ovarian tumor. This study aimed to investigate the effects of genistein on the EGFR mediated signal transduction pathway and to clarify its mechanisms of proliferation inhibition on human ovarian carcinoma cell line SKOV(3) and its xenograft in nude mice.</p><p><b>METHODS</b>The expression of c-erbB-2 protein was determined using immunocytochemistry. The expressions of c-jun and c-fos protein were determined using Western blotting. The expression of c-erbB-2, c-raf-1, c-jun and c-fos mRNA were tested by reverse transcription-polymerase chain reaction (RT-PCR).</p><p><b>RESULTS</b>The expression of c-erbB-2, c-raf-1 and its downstream gene c-jun and c-fos were decreased at mRNA level in the 20 micromol/L genistein group. The expression of c-erbB-2 protein were decreased, its average absorbency (A) were decreased after treatment of SKOV(3) with 20 micromol/L genistein for 48 h, reached at 0.42 +/- 0.02 (P < 0.05). Western blotting demonstrated that the expressions of c-jun and c-fos protein were decreased gradually after being treated with 20 micromol/L genistein for 12 - 72 h.</p><p><b>CONCLUSIONS</b>Genistein could down-regulate the expression of two key genes, c-erbB-2 and c-raf-1 at mRNA and protein level in the EGFR mediated signal transduction pathway, and down-regulate the expression of its downstream nuclear transcription factors c-jun and c-fos at mRNA and protein level. It is suggested that interfering the expressions of some key signal molecules in EGFR mediated signal transduction system by genistein may account for its molecular foundation of proliferation inhibition in ovarian carcinoma.</p>

Effects of genistein on the growth of human ovarian carcinoma transplanted subcutaneously in nude mice / 第三军医大学学报

Objective To investigate the effects of genistein used alone and in combination with cisplatin on the treatment of human ovarian carcinoma transplanted subcutaneously in nude mice. Methods Nude mouse model of human ovarian carcinoma transplanted subcutaneously was established. A total of 30 mice were divided into 5 groups: control group (0 04% saline), genistein group (subcutaneous injection of genistein at 0 2 mg/kg and 0 4 mg/kg), cisplatin group (intravenous injection of cisplatin at 4 mg/kg), and genistein plus cisplatin group. The growth of the subcutaneously transplanted tumor and changes of mouse body weight in each group after treatment using different medication regimens were observed at 7, 14, 21, and 28 d. Histopathological examination was also conducted. Results Genistein (0 4 mg/kg) had significantly inhibitory effect on the transplanted xenograft growth in vivo . Tumor volume, tumor weight, and T/C ratio (mean volume of treated group/mean volume of control group) decreased. Significantly enlarged necrotic areas were found in genistein treated group. The weight loss of nude mice after treatment with genistein was not significantly different from in the control group. Conclusion Genistein has the inhibitory effect on the growth of human ovarian carcinoma transplanted subcutaneously in nude mice. Genistein (0 4 mg/kg) in combination with cisplatin (4 mg/kg) has a synergistic effect on xenograft of human ovarian cancer cell line SKOV 3 in nude mice. The results provide the evidence for the potential usefulness of genistein for the prevention and treatment of human ovarian carcinoma.

Classification of tumors and cultivation of pathologists / 中华医学教育探索杂志

The latest WHO classification of tumors is the most important standard for clinicpathologic diagnosis of pathologists in medical practice.We should study,grasp,and apply it to medical practice for need of patients and ourselves.

Effective activation of CD11c and anti-tumor immune in immunized mice with recombinant E.coli. LLO/OVA / 第三军医大学学报

Objective To investigate the anti-tumor pathway and the effect of recombinant E.coli. LLO/OVA in C57BL/6 mice. Methods Magnetic sorting of splenic CD11c, CD4+ and CD8+ T cell from immunized mice with E.coli. LLO/OVA and E.coli. OVA vaccination were performed. In addition, the lymphocytes mixed reaction and IL-2 and IFN-? in the supernatant were detected. The percentage of OVA257-264 SIINFEKL specific CD8+ T cell were checked by flow cytometry. B16-OVA melanoma protection and inhibition response in E.coli. vaccinated mice were compared. Results Compared to E.coli. OVA, E.coli. LLO/OVA-vaccinated splenic CD11c cells stimulated proliferation of autogenic CD4+ T cells and IL-2 production of these cells. CD11c cells induced autogenic CD8+T cells proliferation and IFN-? secretion. A great number of OVA257-264 SIINFEKL-specific splenic CD8+ T cells were induced and the number of lung tumor nodules was significantly reduced in E.coli. LLO/OVA vaccinated mice. Conclusion Recombinant LLO, as an effective adjuvant of E.coli. OVA, can induce murine splenic CD11c cells activation and play potential roles on CD4+ and CD8+ T cells proliferation and IL-2 and IFN-? secretion of these cells, induce more OVA-specific CD8+ T cells and stimulate stronger tumor inhibition in vivo of vaccinated C57BL/6 mice.

Molecular mechanism of invasion inhibitory effects of genistein on human ovarian serous papillary cystadenocarcinoma cell SKOV_3 / 第三军医大学学报

Objective To investigate how genistein to inhibit the invasion of human ovarian serous papillary cystadenocarcinoma cell line SKOV3.Methods Millicell chamber and coculture method were used to establish chemotactic migration model in vitro to observe the effect of genistein on directional chemotactic migration movement of SKOV3 cells.The protein expressions of cell surface adhesion molecule CD44v6,matrix metalloproteinases and tissue inhibitor of metalloproteinases MMP-9/TIMP-1 and MMP-2/TIMP-2 were determined by using immunocytochemical method and their mRNA levels were examined by in situ hybridization.Results Compared with control group,the directional chemotactic migration of SKOV3 were significantly decreased after treated with 20 ?mol/L and 40 ?mol/L genistein,reached to(46.9?5.8)% and(28.3?4.7)% respectively(P

Anaplastic large cell lymphoma,lymphohistiocytic variant:report of 2 ALK~+ children and review of literatures / 第三军医大学学报

Objective To study the clinicopathological characteristics,diagnosis and prognosis of anaplastic large cell lymphoma,lymphohistiocytic variant(ALCL-LH).Methods Two cases of ALCL-LH were diagnosed according to the WHO standard of lymphoma classification.Their clinical manifestations were analyzed.Lymphoma sample sections were prepared for pathological features and immunohistochemical assays,and examined by in situ hybridization for Epstein-Barr virus(EBV)DNA.Concerning literatures were reviewed.Results Both 2 cases occurred in children.The clinical manifestations presented with fever and enlarged lymph nodes as well as hepatomegaly and splenomegaly.Microscopically,the neoplastic cells,with irregular nuclei,were more often medium to large in size but smaller than those in the common type,and sparsely intermixed with a large number of histiocytes and lymphocytes that exceeded and masked the tumor cell population.The tumor cells clustered around blood vessels.Case 1 showed evidence of erythrophagocytosis.Immunohistochemical stainings showed that tumor cells of 2 cases were positive to CD30,anaplastic lymphoma kinase(ALK),EMA,and Granzyme B,as well as case 1 to CD43,and case 2 to CD7.But both cases were negative to CD45,CD2,CD3,CD4,CD5,CD8,CD45RO,CD56,CD20,CD79a,CD15,MPO,CK,TdT,and LMP1.In addition,the EBV DNA was not detected in the lymphoma.Case 1 and case 2 died 10 d and 3 months after pathological diagnosis respectively.Conclusion ALCL-LH variant is rare subtype,and often occurs in pediatric patients.Morphologically,ALCL-LH is characterized by a few medium to large size cells admixed with a large number of histiocytes and lymphocytes that exceed and mask the tumor cell population.The expressions of CD30,ALK,EMA and cytotoxic granule associated proteins contribute to diagnosis and differential diagnosis.

Growth inhibition effects of artesunate on human cervical cancer cell line HeLa / 第三军医大学学报

Objective To investigate the effects of artesunate on human cervical cancer cell line HeLa.Methods The inhibitory effect on cell proliferation was assessed by MTT assay.Transmission electron microscopy and fluorescent staining were applied to demonstrate the presence of apoptosis.Cell cycle,reactive oxygen species(ROS) levels and mitochondrial membrane potential(??m) were examined by flow cytometry,respectively.Expression of caspase-3 was detected by immunohistochemical methods.Results Growth inhibition of ART on HeLa cells was time-and dose-dependent.Apoptotic feature was observed by transmission electron microscopy and fluorescent staining.The blocking of the cell cycle was in S-phase.The expression of caspase-3 in cytoplasm was positive.The generation of ROS increased obviously(P

Effect of 5-aza-2’-deoxycytidine on MEG3 gene expression and proliferation of human mammary cancer line MCF7 / 中国癌症杂志

Background and purpose:The MEG3 gene is a imprinted gene, whose loss may be associated with the pathogenesis and progression of several tumor types. This study was done to investigate the transcription of MEG3 mRNA in human mammary cancer cell line MCF7 and cell proliferation, in order to explore the effect of the methylation inhibitor, 5-aza-2'-deoxycytidine (5-aza-CdR) on MEG3 gene expression and proliferation in MCF7. Methods:MCF7 was treated with 5 ?mol/L 5-aza-CdR for 2, 4, 6 days, then the alteration of MEG3 gene expression was detected by RT-PCR and Northern blot technology and the proliferation difference in cell growth of MCF7 was observed by MTT. Results:After treated with 5-aza-CdR, the transcription of MEG3 mRNA in MCF7 was increased and the growth of MCF7 was reduced. MCF7 was treated with 5 ?mol/L 5-aza-CdR for 2, 4, 6 days, the inhibitory rates were (23.16?3.93), (49.39?2.38), (64.73?2.24), there were signifi cant differences between them. Conclusion:The growth of MCF7 was possibly inhibited by MEG3 gene, and the downregulation of MEG3 gene might result from the methylation, which was involved in the mammary cancer pathogenesis.

Morphological changes of mitochondria in apoptosis of human ovarian cancer cells induced by arsenic trioxide in vitro / 第三军医大学学报

Objective To investigate the morphological changes of mitochondria and the expression of bcl-2 in the apoptotic SKOV3 and 3AO cells induced by arsenic trioxide (As2O3). Methods Light and electron microscopy, flow cytometry analysis, immunofluorescence flow cytometry analysis were used to detect the apoptotic cells, ultrastructural alteration of mitochondria, and the changes of mitochondrial transmembrane potentials (??m). Results As2O3 induced apoptosis of ovarian cancer cell lines was in a dose dependent manner and various in different cell lines. As2O3 also made a decrease of ??m in SKOV3 and 3AO cells in a dose independent fashion. Electron microscopy indicated that the mitochondria showed swollen, balloon-like appearance and outer membrane disrupted 72 h after As2O3 treatment. Expression of bcl-2 was down-regulated in SKOV3 and 3AO cells after As2O3 treatment. Conclusion The reduce of ??m and down-regulation of bcl-2 may play the key roles in the process of As2O3-induced apoptosis.

Construction of recombinant adenovirus vector expressing EGFP gene specifically in AFP producing liver cancer cells / 中国病理生理杂志

AIM: To construct a recombinant adenovirus vector carrying AFP promoter to specifically express a targeting gene in hepatocellular carcinoma HepG2 cells. METHODS: Based on the Adeno-X~TM Expression system, the CMV promoter was replaced by a 300 bp a-fetoprotein promoter. The EGFP(enhanced green fluorescent protein) gene as a report gene was inserted to the multiple-cloning site(MCS). The normal liver LO2 cells, hepatocellular carcinoma HepG2 cells and HeLa cells were infected by the recombinant adenovirus, respectively. Northern blotting and fluorescence microscope were used to detect the transcription level of EGFP gene and its protein expression, respectively. RESULTS: Northern blotting showed that the target gene was markedly transcribed in HepG2 cells, but slightly in LO2 and HeLa cells. Under the fluorescence microscope, strong EGFP expression was seen in HepG2 cells but very weakly in HeLa and LO2 cells. CONCLUSION: Under the control of the 300 bp human AFP promoter, the target gene carried by the recombinant adenovirus was expressed in the AFP-producing HepG2 cells at a very high level, but not or very weakly in AFP negative cells. This adenovirus system can be used as a new, potent and specific approach for the gene-targeting therapy for the AFP producing primary hepatoma. [

ADENOMATOID TUMORS :AN IMMUNOHISTOCHEMICAL AND ULTRASTRUCTURAL STUDY OF 11 CASES / 重庆医科大学学报

Eleven eases of adenomatoid tumors arc described. There were 9 males and 2 female with an average age of 51 years. These tumors arose in The male and female genital tract. Five of these were located in the epididymis, two in the te- stis, two in the uterus and the last two, one in the spermatic cord and the other in the bladder.The tumor cells of the 11 cases studied were immunoreactive for keratin, but none of these for factor VIII.Only the endothelial cells of the blood vessels in the tumors were positively stained.Ultrastructural examination of 4 cases revealed long brushy microvilli on the surface of the tumor cells lining the lumina. These tumor cells with the presence of tonofilaments in the cytoplasm, were desmosomes.These results strongly support the mesothelial origin of the adenomatoid tumors

Significance of expression of Flt-4 in different histopa thological grades of astrocytoma / 中国病理生理杂志

AIM: To study the significance of expressio n of fms-like tyrosine kinase 4 (Flt-4) in different histopathological grades of a strocytoma. METHODS: The surgical specimens from 50 brain astrocytoma patien ts were stained immunohistochemically for examining Flt-4 and vascular endotheli al growth factor (VEGF) expression. Intratumoral microvessel density (IMVD) was calculated by labeling the endothelial cells of the blood vessels within the tum or. RESULTS: Flt-4, VEGF expression were closely correlated with his topathological grades of astrocytoma. Flt-4 and VEGF expression were found in 52 % (26/50), 60% (30/50) of tumors. A significant correlation was found between Fl t-4 and VEGF expression (P

The study for recombinant E.coli LLO/OVA regulating the function of CD4~+ CD2~+ Treg cells in mice / 中国免疫学杂志

Objective:To investigate the role of recombinant E.coli LLO/OVA on regulating the function of murine CD4+ CD25+ Treg cells.Methods:After E.coli LLO/OVA or E.coli OVA vaccination,the murine spleen CD4+ CD25+ Treg,CD4+ CD25- T and CD11c cells were collected respectively by magnetic beads sorting.The concentration of IL-10 in the supernatant of mix cocultured CD4+ CD25+ Treg and CD11c cells,and the suppression role of CD4+ CD25+ Treg cells on the proliferation of CD4+ CD25- T cells were determined.The percentage of OVA specific CD8+ T cells in mouse spleen was analyzed by flow cytometry.The number of metastatic tumor nodules in lungs of the mice transplanted with B16-OVA subcutaneouly was compared before and after dilition of CD4+ CD25+ Treg cells in mice.Results:Compared to E.coli OVA,E.coli LLO/OVA significantly downregulated IL-10 secretion of CD4+CD25+Treg cells and attenuated the suppressive effect of CD4+ CD25+ Treg on the proliferation of CD4+ CD25- T cells(P

Effect of human ovarian cancer cell apoptosis induced by arsenic trioxide on telomerase activity / 第三军医大学学报

Objective To investicate the effect of apoptosis of human ovarian cancer cell line SKOV_(3) and 3AO exposed to arsenic trioxide on telomerase activity and its mechanisms.Methods The human ovarian cancer cell line SKOV3 and 3OA were treated with arsenic trioxide of different concentration for 12,24,72 h.Cell morphology,PCR-ELISA,RT-PCR were adopted to detect the cell apoptosis and telomerase activity and the expression of human telomerase catalytic subunit(hTERT).Results Arsenic trixide could induce apoptisis of ovarian cancer cell lines,but there exists difference in drug concentration,type of cell line.A dose and time-dependent decline of telomerase activity after SKOV_(3) and 3AO cells exposed to arsenic trixide,meanwhile hTERT mRNA was down-regulated.Conclusion Telomerase activity and hTERT mRNA expression have close relationship and play an important role in arsenic trioxide inducing SKOV_(3) and 3AO cells apoptosis.

Inhibitory effect of genistein on invasion of human ovarian carcinoma cell line SKOV_3 in vivo and in vitro / 第三军医大学学报

Objective To investigate the inhibitory effect of genistein on the invasion of ovarian carcinoma cell line SKOV 3 in vivo and in vitro. Methods The abilities of the genistein-treated SKOV 3 cells to invade through reconstituted matrigel in transwell chambers were investigated in vitro and the invasion effect in vivo was determined using the xenograft models of SKOV 3 in nude mice. Results The ability of the 20 ?mol/L genistein-treated cells to invade the reconstituted basement membrane was decreased significantly at 72 h. This inhibition was in a dose-dependent manner. Genistein at the dose of 40 ?mol/L had the strongest effect. The results in vivo suggested that the grade of invasion in control SKOV 3 cells was in a time-dependent manner and genistein-treated group could apparently inhibit the progress of invasion, localizing the tumor in invasion grade 0 or grade I, and decreasing the proportion of grades Ⅱ, Ⅲ, Ⅳ. Conclusion The results suggest that genistein has inhibitory effect on the invasion of human ovarian carcinoma cell lines SKOV 3 in vivo and in vitro.