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Objective To investigate the importance of hepatitis B virus ( HBV ) infection in the patho-genesis of cholangiocarcinoma ( CC) and further clarify the correlation between the occurrence of intrahepatic and extrahepatic cholangiocarcinoma .Methods HBV protein and gene of 52 formalin fixed,paraffin embedded tis-sues with CC were detected by immunohistochemistry and nest PCR .Results Hepatitis B virus X gene was de-tectable in 33.3%(7/21)of 21 intrahepatic cholangiocarcinoma cases .Hepatitis B surface antigen(HBsAg)was detectable in 21.7%(5/21)and hepatitis B core antigen(HBcAg)was detectable in 19.0%(4/21)of 21 intrahe-patic cholangiocarcinoma cases .In contrast,no HBsAg,HBcAg and hepatitis B virus X gene were detected in any of the 31 extrahepatic cholangiocarcinoma cases .Conclusion HBV infection is a significant risk factor for intra-hepatic cholangiocarcinoma ,but not for extrahepatic cholangiocarcinoma ,in Northeast of China .The integration of HBV gene may be involved in the carcinogenesis of intrahepatic bile duct epithelial cells .
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Objective To evaluate the effects of malignant ascites on the morphological characteristics and the proliferation and migration abilities of the tumor cell and ovarian cancer cell lines(SKOV3)in the ovarian cancerous ascites.Methods Tumor cells extracted from the ovarian cancerous ascites were cultured in vitro with DMEM high glucose culture medium,and ovarian cancer cell lines( SKOV3) were cultured in DMEM high glucose and DMEM high glucose with different proportion of malignant ascites.The morphological characteristics of the cells were observed by optical microscope and electron microscope respectively.Cell proliferation ability was de-tected by CCK kit;The effect of SKOV3 on the migration of ovarian cancer cell lines was measured by scratch test.Results The morphological characteristics of tumor cells and ovarian cancer cell lines( SKOV3) in ovarian cancer ascites were significantly different.The proliferation ability of tumor cells was decreased without the asci-tes.The proliferation and migration abilities of SKOV3 cultured in mixed culture medium were significantly im-proved compared with the cells cultured in high glucose medium.Conclusion The change of tumor cell morphol-ogy in ascites benefits its abilities of proliferation and migration.The malignant ascites promote the abilities of pro-liferation and migration of ovarian cancer cell line(SKOV3).
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Objective To explore the relationship between HPV infection ,the expression of PCNA ,Ki-67,p53 in cervical carcinoma and clinicopathologic data .Methods Seventy-six cases of cervical cancer tissues with complete clinical data were collected from the third affiliated hospital of harbin medical university from march 2012 to September 2012 .Tissue microarray technology ,immunohistochemical technique and statistic method were adopted to make comparative analysis among HPV ,PCNA,Ki-67,and p53 expression in patrents with cervical cancer.Results The expression rate of HPV in cervical carcinoma was 68.4%,which was related to the lym-phatic metastasis(P<0.05).Expression of PCNA in cervical cancer tissue was 90.1%and 100%in female pa-tionts women under the age of 35.The expression rate of Ki -67 and p53 were 48.9%and 38.2%respectively, both were correlated to lymphatic metastasis(P<0.05)and preoperative chemotherapy(P<0.01).Conclusion HPV infection with Ki -67,and p53 expression in cervicalcancer tissues are associated with the presence of lymph node metastasis .HPV promotes cervical epithelial cell proliferation ,inactivates tumor suppressor gene p 53 function ,and promotes the invasion and metastasis of cervical cancer .Preoperative chemotherapy of cervical canc-er can reduce the expression of p53,PCNA and Ki-67.
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BACKGROUND: As for the treatment of chylous fistula concurrent in oral-maxillofacial tumor resection simultaneously undergoing neck lymph node dissection, many different approaches have been put forward. A simple surgical ligation, strong negative pressure drainage, filling the muscle tissue alone or a combination of the above methods are all unsatisfactory regarding the prognosis and curative effect.OBJECTIVE: To evaluate the validity of medical biological adhesive cohering peripheral autologous muscle tissues to block thoracic duct fistula in order to prevent chylous fistula following neck lymph node dissection.METHODS: All of the 12 patients were detected and diagnosed as chylous fistula in neck lymph node dissection surgery, the wounds were immediately sutured and treated with medical biological adhesive cohering peripheral autologous muscle tissues to block thoracic duct fistula. RESULTS AND CONCLUTION: Of all the 12 patients, 10 recovered without chylous fistula or severe complications, and reoperations were adopted to cure the failed 2 cases. All patients were visited 3 months postoperatively, no recurrence of chylous fistula, local stimulus response or allergy was found. It is suggested medical adhesive to block thoracic duct fistula may be an effective and safe way for prevent chylous fistula following neck lymph node dissection.
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BACKGROUND:Recently,acellular dermal matrix allograft has been widely used in the repair of oral mucosal defects.But little information is about the heterogeneous acellular dermal matrix (HADM) patch for repair of oral mucosal defects.OBJECTIVE:To investigate the efficacy and biosafety of HADM in the repair of oral mucosal defects.METHODS:In total 71 patients with oral benign or malignant tumors who had oral mucosal or soft tissue defects following tumorectomy were included in this study.These patients comprised 37 males and 34 females,and were averaged 45 years (range,20-70 years old).Of them,42 suffered from benign tumors and 29 from malignant tumors.HADM patches were used for repair of oral mucosal defects.The survival,color,and texture of HADM patches were observed.Shrinkage rate of HADM patches was compared between regions without supports from hard tissues (cheeks,tongue,and mouth floor) and with supports from hard tissues (gingiva,hard palate).RESULTS AND CONCLUSION:All 71 HADM completely survived.No necrosis and infection occurred.At 2 weeks after transplantation,(98.20±5.20) % of patch area survived.At 3 months after transplantation,patches showed similar color to surrounding oral mucosa and most patients had sense of tension to different extents.At 6 months after transplantation,cell creeping substitution and vasculadzation were successfully accomplished in the region of patch transplantation.Patches grew stably,with smooth pink appearance and good elasticity,and no further shdnkage.Patients felt normal.HADM patch shrank primarily at 2 weeks-1 month after transplantation,and tended to be stable at 3 months.There was no significant difference in tissue morphology between surgical region and normal tissue.The HADM shdnkage rate was significantly higher in regions without supports from hard tissues than regions with supports from hard tissues.These findings indicate that HADM patches have advantages in repair of oral mucosal defects including good histocompatibility,wide source,simple manipulation,and able to cover the wound surface in the early state,promote wound surface healing,and reduce scar formation,and can be used as an ideal matedal for repair of oral mucosal defects.
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ACKGROUND: Adriamycin (ADM) can specifically conjugate with receptor. In particular, ADM nanopartides play a target role in decreasing the cytotoxicity.OBJECTIVE: To investigate the targeting effect of addamycin nanoparticles conjugated with hyaluronic acid (ADM-HA-SSL) on oral squamous carcinomas calls in vitro.DESIGN, TIME AND SETTING: An in vitro contrast observation was performed in College of Madne Life Science, Ocean University of China from January to July 2008. MATERIALS: Oral squamous carcinomas calls were sincerely presented by Professor Chen from the Ninth People's Hospital of Shanghai; ADM-HA-SSL (drug loading 156 mg/L) was sincerely presented by Professor Liu from College of Marine Life Science, Ocean University of Chine.METHODS: Oral squamous carcinomas cells were cultured with 0.5, 1.0, 5.0, and 10.0 mg/L ADM-HA-SSL. MTT assay was used to detect the targeted cytotoxicity of ADM-HA-SSL against oral squamous cell carcinomas. With the concentrations of 5.0 and 10.0 mg/L, call apoptosis was ascertained by call flow cytometry after 6, 12, 24 and 48 hours. MAIN OUTCOME MEASURES: Cell survival rate and apoptosis rate.RESULTS: At 24 and 48 hours after induction, cytotoxicity assay revealed that the effect of ADM-HA-SSL was superior to that of free ADM (t=5.78-42.05, P < 0.01). The results of flow cytometry showed that the apoptosis rate was enhanced with the increase of the time (F=4 200.40, 4 775.36, P < 0.01), and the rate was also increased at the same time point with the increase ofconcentration (t=12.06-20.08, P < 0.05).CONCLUSION: ADM-HA-SSL can specifically recognize oral squamous carcinomas cells and deliver adriamycin into the cells. And the effect is enhanced by the time prolonging.
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With the help of computer image analysis system, we used the method of average positive stained area percentage APSAP to evaluate the slice immunohistochemistry result. Then we compared the evaluation result with the result of manual counting. Conformity between the two methods was verified. These data indicated that the method of was in accord with manual counting to a great extent. Moreover, the theory basis, advantages and disadvantages of the method were discussed in this paper.
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Humans , Image Interpretation, Computer-Assisted , Methods , Immunohistochemistry , Methods , Staining and Labeling , Methods , Stomach Neoplasms , Chemistry , PathologyABSTRACT
BACKGROUND: Many operations for isolating, purifying and identifying bone marrow mesenchymal stem calls (BMSCs) are complicated and cost much. Also they have great effect on cell activity. Whether whole bone marrow adherent culture can avoid above-mentioned disadvantages remains unclear. At present, many studies huve been done to confirm an effective and low cost method for isolating, purifying and identifying such cells.OBJECTIVE: This study is to in vitro induce and differentiate rat BMSCs by whole bone marrow adherent culture,and to identify the cells.DESIGN: A controlled observational experiment.SETTING: Qingdao University Medical College.MATERIALS: This study was carried out in the Laboratory of Oral Cavity and Laboratory of Molecular Biology (provincial level) Qingdao University Medical College between November 2005 and March 2007. Twenty Wistar rats of either gender, aged 3 to 4 weeks, of SPF grade, weighing 120-150 g, were provided by the Qingdao Laboratory Center. The protocol was carried out in accordance with animal ethics guidelines for the use and care of animals. Fetal bovine serum (FBS, Hangzhou Sijiqing Bioengineering Material Research Institute), alkaline phosphatase (ALP) kit (Nanjing Jiancheng Bioengineering Research Institute), reverse transcription kit (American Promega Corporation) and primer (Shanghai Bioengineering Co.,Ltd.) were used in this study.METHODS: Adult rat BMSCs were isolated and cultured by whole bone marrow adherent culture. They were digested with 2.5 g/L trypse and inoculated at a density of 5 ×107 L-1 in 6-well culture plate. Then, the cells were divided into experimental group and control group. Inducing culture medium was added to experimental group, and the same amount of basic culture medium was added to control group. ① Cell differentiation and calcium tuberculation were observed under the inverted microscope. ② Biological characteristics of induced cells were detected by calcium tubercle Von Kossa and alizarin Bordeaux. ③ALP activity was detected by diazo salt staining. ④Human core binding factor alpha subunit-1 (Cbf α-1), osteocalcin (OCN) and osteoblast-specific Osterix (OSX) mRNA expressions were detected by reverse transcription-polymerase chain reaction (RT-PCR).MAIN OUTCOME MEASURES: ① Induction and differentiation results of cells. ② Biological characteristics of cells induced by rat BMSCs. ③ ALP activity. ④ Cbf α-1, OCN and OSX expressions.RESULTS: ①Inducing culture medium was added in the serial subcultivation. About 9 days later, cell clones were connected to each other. On about 21 to 28 days, some pykno-round mineralized tubercles appeared. Meanwhile,control cells were connected to each other, but they did not form the tubercle. ② In the experimental group, when MSCs were induced for 21 to 28 days, obvious round or oval calcified tubercles were seen by naked eyes. The results of Von Kossa staining exhibited black sediments, and those of alizarin Bordeaux staining exhibited salmon tubercles. Calcium tubercles were not found in the control group. ③The ALP activity after 2 weeks of induction was obviously increased in the experimental group, but was relatively weak in the control group. ④In the experimental group,Cbf α-1, OCN and OSX expressions were significantly increased after induction.CONCLUSION: After being in vitro induced and differentiated by whole bone marrow adherent culture, rat BMSCs exhibited morphological and biological characteristics similar to typical osteoblasts.
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Objective To study the relationship between pathological, immunohistochemical and ultrastructural changes of skeletal myodystrophy (SMD) and the development of the disease.Methods SMD tissue of 20 cases were routinely processed,the paraffin sections,the semi thin sections and the ultrasthin sections were observed by light microscopy and electron microscopy.Results 20 cases with SMD tissue were divided into three groups: Simple SMD for 8 cases, major changes were regional; Progressive SMD for 10 ca ses, the pathological changes were diffuse with large amount of degeneration of cell organs; SMD derived from nerve injury for 2 cases, pathological changes of the part controlled by the nerve were observed. While SMD was injured, myosin got deneration first.Conclusion The pathological and ultrastructure changes could be used to judge the progressive degree of myodystrophin. The amount of lost myosin could forecast the progression of the disease.
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Objective: To determine the relationship of the expression of transforming growth factor ?1(TGF ?1), TGF ?2 and TGF? receptor(T?RⅡ) with the development of squamous cell carcinoma (OSCC). Methods: 40 cases of OSCC and 20 cases of normal oral mucosa were examined with SABC immunohistochemical staining for the expression of TGF ?1, TGF ?2 and T?RⅡ. Results: Semi quantitative analysis revealed that TGF ?1, TGF ?2 and T?RⅡ were expresed in OSCC and normal oral mucosa. TGF ?1, TGF ?2 were overexpressed in OSCC compared with those in normal oral mucosa, while T?RⅡ expression were reduced. Overexpression of TGF ?1, TGF ?2 and reduced expression of T?RⅡ were associated with pathological grade, clinical stage and neck lymph node metastasis( P
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Objective: To detect the role of hypoxia in the regulation of vascular endothelial growth factor (VEGF) and matrix metalloproteinase 2 (MMP 2) in oral squamous cell carcinoma cell lines. Methods: Oral squamous cell carcinoma TSCCa and TNM cells were treated by hypoxia at ?=1% of oxygen for 4～12 h. VEGF and MMP 2 in the cells were quantitatively determined by enzymelinked immunosorbent assay (ELISA). The mRNA expression of VEGF and MMP 2 in the two cell lines was evaluated by semi quantitative RT PCR. Results: In both cell lines VEGF and MMP 2 were increased after 4 hour hypoxia treatment (P
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Objective: To study the role of transforming growth fa ctor ?1(TGF-?1) in the procession of human oral squamous cell carcinoma. Methods: Immunohistiochemical techinic was used to determine o ncogene c-myc,c-erbB2,TGF-?1 and EGFR protein expression in oral squamou s cell carcinoma cell line TSCCa and metastasis carcinoma cell line GNM treated by different levels of TGF-?1. Results: TGF-?1 showed different effects on the two cell lines. TGF-?1 significantly increased the expression of C-myc,CerbB2 and slightly enhanced TGF-?1 and EGFR protein expression in GNM cells, while TGF-?1 decreased the expression of C-myc,C erbB2,TGF-?1 and EGFR in TSCCa cells; moreover, the changes of protein ex pression of C-myc,CerbB2 were dose-dependent on TGF-?1. Conclusi on: TGF-?1 have different role in the oral squamous cell carcinoma cell lines.
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Objective: To study the significance of intracellular free calcium levels ([Ca 2+]i) and protein kinase C(PKC) levels in the process of human salivary gland mucoepidermoid carcinoma MEC-1 cell apoptosis induced by adriamycin. Methods: MEC-1 cells were treated with adriamycin at 10 ?mol/L for 30 s～24 h.Apoptosis of the cells was investigated by light and electron microscopy, agarose gel electrophoresis and flow cytometry. [Ca 2+]i was determined by flow cytomerty, PKC by Bardford method. Results: The results showed that MEC-1 cells presented classic morphologic features of apoptosis. [Ca 2+]i in the treated cells was increased from (36.63?0.61) nmol/L to (84.00?0.45) nmol/L after 30 s～24 h treatment,while that in the control cells was 17.43?0.47 (P
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Objective To study the effects of P53, PCNA, Bcl-2 protein and their relationship in salivary adenoid cystic carcinoma(SACC). Methods These proteins were examined by immunohistochemistry. Results Overexpressions of P53 and PCNA were revealed in ACC samples, they were higher than those in (polymorphous adenomas) PA, but expression of Bcl-2 protein was not different between ACC and PA. In 3 subtypes of ACC, expressions of 3 proteins were different. Conclusion Mutations of P53, Bcl-2 may be involed in the occurrence of SACC, expression of PCNA and mutation of P53 may coexist in the development of the SACC.
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Objective To explore whether or not driamycin inhibits mEC cells by apoptosis. MethodsAdriamycin was used for mucoepidermoid carcinoma cells mEC-l. Morphologic changes were observed atdifferent times by light and electronic microscope. DNA fragments were shown on agarose gelelectrophoresis. DNA content and cell cycles were analyzed by flow cytometer. Changes of intracellularcalcium ion was monitored by flow cytometer. Results mEC-1 cells presented with classic morphologicfeatures of apoptosis. Intracellular calcium ion was increased. Conclusion Apoptosis followed by thechanges of intracellular calcium ion in human salivary gland mucoepidermoid carcinoma cells is one of themechanisms of adriamycin inhibited tumor cell growth.