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1.
Acta Physiologica Sinica ; (6): 244-252, 2021.
Article in Chinese | WPRIM | ID: wpr-878253

ABSTRACT

The aim of this study was to investigate the effects of polarization program on the ability of macrophages to regulate iron metabolism. M1 and M2 macrophages were propagated in vitro from porcine alveolar macrophages 3D4/2 and polarized by cytokines. The 3D4/2 macrophages were treated with 20 ng/mL interferon gamma (IFN-γ) and 10 ng/mL interleukin-4 (IL-4) combined with 10 ng/mL macrophage colony-stimulating factor (M-CSF) to induce polarization to M1 and M2, respectively. After incubation for 24 h, the expression levels of inflammatory factors and iron-metabolism genes were determined using real-time qPCR, Western bot and immunofluorescence. The M1/M2 macrophages culture media supernatant was collected and used to treat porcine intestinal epithelial cells IPEC-J2. The proliferation ability of IPEC-J2 was detected using CCK-8 assay kit. Following exogenous addition of ammonium ferric citrate (FAC) to M1/M2 macrophages, the phagocytic function of macrophages was detected using fluorescein isothiocyanate-dextran (FITC-dextran) and flow cytometry. The results showed that, compared with control, M1 macrophages had higher mRNA levels of iron storage proteins (ferritin heavy and light polypeptide, i.e. FtH and FtL), hepcidin and lipocalin-2, as well as iron content. Moreover, iron enhanced the ability of M1 macrophages to phagocytize FITC-dextran. There was no significant change in these mRNA expression levels in M2 macrophages, but the mRNA expression levels of ferroportin and transferrin receptor were up-regulated. In addition, the conditioned media supernatant from M2 macrophages promoted cell proliferation of IPEC-J2. These findings indicate that M1 macrophages tend to lock iron in the cell and reduce extracellular iron content, thereby inhibiting the proliferation of extracellular bacteria. While M2 macrophages tend to excrete iron, which contributes to the proliferation of surrounding cells and thus promotes tissue repair.


Subject(s)
Animals , Cytokines , Ferritins , Iron/metabolism , Macrophages/metabolism , Macrophages, Alveolar/metabolism , Swine
2.
Acta Academiae Medicinae Sinicae ; (6): 945-949, 2021.
Article in Chinese | WPRIM | ID: wpr-921563

ABSTRACT

Insomnia is a subjective experience of difficulty in falling asleep and/or maintaining sleep accompanied by the impairment of daytime social functioning due to insufficient sleep quality or quantity to meet normal physiological needs.It has chronic damage to all the human body systems and is the most common sleep disorder.The main mechanism for the occurrence and maintenance of insomnia is the hyperarousal hypothesis,and microarousal,as a cortical arousal,is also involved in the formation of the hyperarousal mechanism.The mechanism and clinical significance of microarousal were reviewed and summarized in this paper in order to guide the clinical work.


Subject(s)
Humans , Arousal , Sleep , Sleep Initiation and Maintenance Disorders , Sleep Quality
3.
Chinese Journal of Ocular Fundus Diseases ; (6): 400-403, 2020.
Article in Chinese | WPRIM | ID: wpr-871761

ABSTRACT

Nanophthalmos is a congenital disorder, which is characterized by small eye without any ocular or systemic malformation. It has autosomal-dominant and recessive forms of inheritance. It manifests as reduced total axial length, high hyperopia and thickened sclera. Nanophthalmos is relatively rare, but at high risk of secondary angle-closure glaucoma, uveal effusion syndrome. It can be spontaneous remission or recurrent, and responses poorly to medication. The effect of systemic glucocorticoid treatment is not ideal. The surgery challenges with very high rate of intra- and post-operative complications such as hemorrhage, choroidal and retinal detachment, malignum glaucoma and uveal effusion, so that the prognosis is poor. It is of great instructive significance to further understand the clinical features, complications and treatment progress and to avoid the missed diagnosis and misdiagnosisi of its complications, and thus choose the right management.

4.
Recent Advances in Ophthalmology ; (6): 979-983, 2017.
Article in Chinese | WPRIM | ID: wpr-657798

ABSTRACT

Age-related macular degeneration (AMD) is the main cause of blindness in the people older than 50 years old,and atrophic age-related macular degeneration accounts for 85%-90% patients of AMD.Following the application of multi-spectral imaging,fundus autofluorescence,optical coherence tomography,microperimetry,multifocal electroretinogram and other new methods in ophthalmic clinical,the morphological and functional changes of atrophic AMD lesions have been more in-depth and comprehen sive,so this article will give a review on the relevant progress of examination techniques in recent years.

5.
Recent Advances in Ophthalmology ; (6): 979-983, 2017.
Article in Chinese | WPRIM | ID: wpr-660238

ABSTRACT

Age-related macular degeneration (AMD) is the main cause of blindness in the people older than 50 years old,and atrophic age-related macular degeneration accounts for 85%-90% patients of AMD.Following the application of multi-spectral imaging,fundus autofluorescence,optical coherence tomography,microperimetry,multifocal electroretinogram and other new methods in ophthalmic clinical,the morphological and functional changes of atrophic AMD lesions have been more in-depth and comprehen sive,so this article will give a review on the relevant progress of examination techniques in recent years.

6.
China Journal of Chinese Materia Medica ; (24): 603-608, 2016.
Article in Chinese | WPRIM | ID: wpr-230110

ABSTRACT

To investigate the effects of Liu Tea extracts(LTE) on proliferation, apoptosis and drug sensitivity of drug-resistant gastric cancer cell BGC823/5-FU. MTT assay was used to analyze effect of LTE on cell growth and sensitivity chemotherapeutic drugs, and synergistic effect of the combination of LTE with 5-FU on BGC823/5-FU cells. Combination index (CI) was calculated by CompuSyn. Cell apoptosis was measured by flow cytometry (FCM). Protein expressions of P-gp, Bcl-2, Bax and Caspase-3 (17KD) were detected by Western blot at different concentrations of LTE in BGC823/5-FU cells (100, 200, 400 mg•L⁻¹). The results showed that LTE had an inhibitory effect on growth of BGC823/5-FU cell in a dose-time-dependent manner and significantly reduced IC₅₀ of 5-FU, CDDP, PTX and ADM to BGC823/5-FU cells(P<0.05), indicating it could reverse tolerance of drug resistant cells to chemotherapeutic drugs, with reversion multiples of 2.35, 1.68, 1.96, 0.52. The combination of LTE with 5-FU had positive synergistic effect on the BGC-823 cell line. FCM assay suggested that LTE could induce BGC823/5-FU apoptosis. The apoptosis rate was up to 46.2% when the cells were treated with 800 mg•L⁻¹ LTE after 24 h(P<0.01). According to the protein detection results, with the increase in concentration of LTE, the protein expression of Bcl-2 was gradually decreased(P<0.01), the expression of Bax and Caspase-3 were extremely increased(P<0.01), with statistical significance in difference(P<0.01) but no difference in the expression of P-gp between experiment group and control group. LTE can inhibit the growth of drug-resistant human gastric cancer cell BGC823/5-FU and reverse its chemotherapeutic tolerance to some extent. Inhibition of antiapoptotic proteins, activation of proapoptotic proteins and induction of apoptosis of resistant cells may be its main mechanisms.

7.
Chinese Journal of Oncology ; (12): 804-807, 2013.
Article in Chinese | WPRIM | ID: wpr-267451

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effects of a Chinese herbal extract Songyou Yin on residual hepatocellular carcinoma after chemotherapy in nude mice and the relevant mechanisms.</p><p><b>METHODS</b>Orthotopic nude mouse models bearing residual hepatocellular carcinoma after chemotherapy was established using human liver carcinoma MHCC97L cells. Three different doses of Songyon Yin (2.1 g/kg, 4.2 g/kg and 8.4 g/kg) were administered to the mice in the trial groups by intragastric gavage, respectively. The mice in the control group were administered physiological saline. The tumor growth, metastasis and survival in the mice of each group were recorded. The corresponding mechanisms were studied.</p><p><b>RESULTS</b>The pulmonary metastasis rates of the control group and 2.1g/kg, 4.2g/kg, 8.4g/kg Songyou Yin treatment group were 86.7%, 73.3%, 40.0%, and 20.0%, respectively, and the survivals of these groups were 53.83 ± 4.71, 56.50 ± 6.09, 66.67 ± 5.61, 81.17 ± 7.36 days, respectively. Compared with the mice in the control group, mice in the 4.2 g/kg, 8.4 g/kg Songyou Yin treatment groups had a lower pulmonary metastasis rate (P = 0.021 and P = 0.001, respectively) and longer survival (P = 0.002 and P = 0.001, respectively). A restoration of E-cadherin expression and a concomitant reduction of N-cadherin expression were detected in the tumors of the 4.2 g/kg and 8.4 g/kg Songyou Yin treatment groups.</p><p><b>CONCLUSIONS</b>Songyou Yin effectively inhibits the invasion and metastasis of the residual hepatocellular carcinoma after chemotherapy in nude mice through attenuating the epithelia-mesenchymal transition and prolongs the survival. Songyon Yin may have potential to promote the efficacy of chemotherapy in hepatocellular carcinoma.</p>


Subject(s)
Animals , Humans , Male , Mice , Antineoplastic Agents , Therapeutic Uses , Antineoplastic Agents, Phytogenic , Pharmacology , Cadherins , Metabolism , Carcinoma, Hepatocellular , Drug Therapy , Metabolism , Pathology , Cell Line, Tumor , Drug Combinations , Drugs, Chinese Herbal , Pharmacology , Epithelial-Mesenchymal Transition , Liver Neoplasms , Drug Therapy , Metabolism , Pathology , Lung Neoplasms , Mice, Inbred BALB C , Mice, Nude , Neoplasm Transplantation , Neoplasm, Residual , Metabolism , Pathology , Organoplatinum Compounds , Therapeutic Uses , Plants, Medicinal , Chemistry , Random Allocation , Survival Rate , Tumor Burden , Xenograft Model Antitumor Assays
8.
Chinese Journal of Hepatology ; (12): 604-608, 2013.
Article in Chinese | WPRIM | ID: wpr-278031

ABSTRACT

<p><b>OBJECTIVE</b>To construct a high metastatic potential human hepatocellular carcinoma (HCC) orthotopic transplantation model with palliative liver resection in nude mice.</p><p><b>METHODS</b>A human HCC orthotopic nude mice model was established by administering a single inoculation of the highly metastatic MHCC97H tumor tissue (size 2 mm * 2 mm * 2 mm) into the left liver lobe. At day 14 post-inoculation, a random group of the mice received palliative liver resection; the unresected mice served as controls. Changes in expression levels of 113 genes with metastasis-related functions were evaluated in the residual HCC tissues. At day 35 post-resection, a random group of the mice were sacrificed by cervical dislocation and a comprehensive metastases examination was performed. The remaining mice were used to observe life span. All statistical analyses were performed by the SPSS v17.0 software, and significance was defined as P less than 0.05.</p><p><b>RESULTS</b>The nude mouse model of highly metastatic HCC with palliative liver resection was successfully established. Incidences of intrahepatic and abdominal metastases were higher in the palliative resected group (vs. unresected group: 11.7+/-4.7 vs. 6.3+/-2.8, t = -2.412, P less than 0.05 and 9.8+/-3.4 vs. 5.2+/-2.6, t = -2.641, P less than 0.05 respectively). In addition, the palliative resected group showed significantly enhanced pulmonary metastasis (vs. unresected group: 14.3+/-4.7 vs. 8.7+/-4.7, t = -2.348, P less than 0.05). Differential gene expression levels were found for MTSS1, TGFbl, SMAD2, IL-1b, and MMP7, and were situated in the central position of gene function net of residual HCC. The life-span of the palliative resected group was significantly longer than that of the unresected group (60.8+/-2.7 vs. 51.3+/-1.4 days, x2 = 12.850, P less than 0.01).</p><p><b>CONCLUSION</b>The highly metastatic human HCC nude mouse model with palliative liver resection that was successfully constructed in this study represents a useful investigational tool to assess the biological characteristics of residual cancer and to screen therapeutic strategies.</p>


Subject(s)
Animals , Humans , Mice , Carcinoma, Hepatocellular , Pathology , General Surgery , Disease Models, Animal , Hepatectomy , Liver Neoplasms, Experimental , Pathology , General Surgery , Mice, Inbred BALB C , Mice, Nude , Neoplasm Metastasis , Neoplasm Transplantation , Tumor Cells, Cultured
9.
Chinese Journal of Oncology ; (12): 805-809, 2012.
Article in Chinese | WPRIM | ID: wpr-307289

ABSTRACT

<p><b>OBJECTIVE</b>To explore the changes of metastatic potential of residual hepatocellular carcinoma (HCC) after in vivo chemotherapy and its mechanism.</p><p><b>METHODS</b>Nude mouse models of orthotopic HCC in the nude mouse livers was established using human hepatocellular carcinoma cell line MHCC97L cells. Oxaliplatin (10 mg/kg, once per week) was administered intraperitoneally (i.p.) to mice in the trial group. Mice in the control group received 0.2 ml of 0.9% sodium chloride on the same days. On day 7 after the third injection, all mice were sacrificed and tumor fragments of equal volume (2 mm×2 mm×2 mm) from each mouse of the oxaliplatin-treated and untreated groups were reinoculated into the livers of each new recipient mouse correspondingly. The growth, metastasis and molecular phenotype of the reinoculated tumors in both groups were determined.</p><p><b>RESULTS</b>In the new recipient mice, compared with untreated tumors, oxaliplatin pre-treated tumors grew significantly slower [(2624.59 ± 491.60) mm(3) vs. (3849.72 ± 827.09) mm(3), P < 0.001], but gave more spontaneous metastasis to the lung (10/12 vs. 3/12, P = 0.012). A decreased expression of E-cadherin and increased expression of N-cadherin, vimentin and transcription factor Snail were detected in the oxaliplatin pre-treated tumors by immunohistochemistry, which provided the evidence of epithelial mesenchymal transition (EMT) in these tumors.</p><p><b>CONCLUSION</b>Residual hepatocellular carcinomas after in vivo chemotherapy grow slower but gain enhanced metastatic potential to the lung, associated with epithelial mesenchymal transition.</p>


Subject(s)
Animals , Humans , Male , Mice , Antineoplastic Agents , Therapeutic Uses , Apoptosis , Cadherins , Metabolism , Carcinoma, Hepatocellular , Drug Therapy , Metabolism , Cell Line, Tumor , Epithelial-Mesenchymal Transition , Liver Neoplasms , Drug Therapy , Metabolism , Pathology , Lung Neoplasms , Drug Therapy , Mice, Inbred BALB C , Mice, Nude , Neoplasm Metastasis , Neoplasm Transplantation , Neoplasm, Residual , Drug Therapy , Metabolism , Organoplatinum Compounds , Therapeutic Uses , Snail Family Transcription Factors , Transcription Factors , Metabolism , Tumor Burden , Vimentin , Metabolism , Xenograft Model Antitumor Assays
10.
Chinese Journal of Oncology ; (12): 344-349, 2010.
Article in Chinese | WPRIM | ID: wpr-260402

ABSTRACT

<p><b>OBJECTIVE</b>The aim of this study was to evaluate the correlation of protein expressions of CXC chemokine receptor 4 (CXCR4), vascular endothelial growth factor-C (VEGF-C) and cytokeratin 19 (CK-19) with lymph node metastasis (LNM) in patients with hepatocellular carcinoma (HCC), and their survival.</p><p><b>METHODS</b>The expressions of CXCR4, VEGF-C and CK-19 in HCC patients with (n = 123) or without (n = 145) LNM were determined using tissue microarray and immunohistochemical staining. The relationship between clinicopathological features and CXCR4, VEGF-C and CK-19 were analyzed. Evaluation of immunostaining was performed semiquantitatively by visual assessment.</p><p><b>RESULTS</b>The UICC T stage, and expressions of nuclear CXCR4, VEGF-C and CK-19 were independent risk factors for LNM. Nuclear CXCR4, VEGF-C and CK-19 expression were predictive factors for LNM in HCC patients. In patients with LNM, the median survival time was 15.1 months for patients with high nuclear CXCR4 expression and 24.5 months for those with low nuclear CXCR4 expression. The median survival time was 15.1 months for patients with high tumor VEGF-C expression and 31.1 months for those with low tumor VEGF-C expression. The median survival time was 12.0 months for patients with positive CK-19 expression and 19.2 months for patients with negative CK-19 expression. Patients with high nuclear CXCR4, VEGF-C or CK-19 expression had significantly poorer prognosis than those with low expression (all P < 0.05). PVT, UICC T stage and expressions of nuclear CXCR4, VEGF-C, and CK-19 were independent prognostic factors.</p><p><b>CONCLUSION</b>Increased protein expressions of nuclear CXCR4, VEGF-C, and CK-19 are independent risk factors for developing lymph node metastasis, and they are significantly correlated with LNM and poor outcome in HCC patients.</p>


Subject(s)
Female , Humans , Male , Middle Aged , Carcinoma, Hepatocellular , Metabolism , Pathology , Cell Nucleus , Metabolism , Follow-Up Studies , Keratin-19 , Metabolism , Liver Neoplasms , Metabolism , Pathology , Lymphatic Metastasis , Pathology , Neoplasm Staging , Proportional Hazards Models , Receptors, CXCR4 , Metabolism , Risk Factors , Survival Rate , Vascular Endothelial Growth Factor C , Metabolism
11.
Chinese Journal of Hepatology ; (12): 915-919, 2010.
Article in Chinese | WPRIM | ID: wpr-360798

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effects of lentivirus mediated siRNA targeting human metastasis suppressor 1 (MTSS1, MIM-B gene) gene on the invasive and metastatic potentials of hepatocellular carcinoma (HCC) MHCC97H cells.</p><p><b>METHODS</b>The siRNA targeting MTSS1 was cloned into one lentivirus work vector. The work vector and three package plasmids were co-transfected into 293T cells with the help of lipefeetamine 2000. Lentivirus was collected in 72 hours and was added to the cultured MHCC97H cells. The total cell MIM-B mRNA and MIM-B protein were extracted and underwent real-time PCR and western-blot test respectively. Boden chamber assay was used to evaluate the invasive potential of MHCC97H cells. Gelatin zymography was used to detect matrix metalloproteinase-2 (MMP2) activity. Metastatic human HCC nude mice models were established by orthotopic implantation with a high metastatic potential human HCC cell line MHCC97H. Twenty-four nude mice bearing orthotopic xenografts were randomized into black control group, Lenti-GFP group and intervention group (Lenti-MTSS1 group) 14 days after orthotopic implantation (8 per group). The ultrasound-guided multi-point injection was performed on mice with borate buffered saline, Lenti-GFP and Lenti-MTSS1 respectively. Mice were sacrificed on day 35 for the examination of pulmonary metastasis. The SPSS 13.0 soft ware was applied to data analysis.</p><p><b>RESULTS</b>The small interfering RNA targeting MTSS1 was constructed successfully with a transfection efficiency of 97.0%, which produced a marked inhibition of invasive ability of MHCC97H cells through Matrigel, being 37.9+/-4.4, 37.4+/-5.3 and 26.6+/-4.6 in the black control group, Lenti-GFP group and Lenti-MTSS1 group (F = 26.695, P value is less than 0.01), respectively. MIM-B expression and MMP2 activity of intervention group were also significantly down-regulated as compared to the control group. The results of in vivo studies showed that the numbers of lung metastatic nodules were 6.5+/-2.6, 6.4+/-2.7 and 3.8+/-1.3 in the black control group, Lenti-GFP group and intervention group respectively with significant statistical difference (F = 3.637, P value is less than 0.05), accorded with tumor tissue MIM-B mRNA expression of 0.39+/-0.19, 0.38+/-0.10 and 0.16+/-0.11 respectively (F = 11.644, P value is less than 0.01) when comparison was made between control group and therapy group.</p><p><b>CONCLUSION</b>Small interfering RNA mediated by lentivirus inhibited MIM-B expression and resulted in inhibition of the invasive and metastatic potentials of MHCC97H cells, which may attributed, in part, the down regulation of MMP2 activity, and thus may provide a new molecular targeted therapy for HCC patients in the future.</p>


Subject(s)
Animals , Humans , Mice , Carcinoma, Hepatocellular , Genetics , Pathology , Cell Line, Tumor , Matrix Metalloproteinase 2 , Metabolism , Mice, Inbred BALB C , Mice, Nude , Microfilament Proteins , Genetics , Neoplasm Proteins , Genetics , Neoplasm Transplantation , RNA, Small Interfering , Genetics , Transfection
12.
Chinese Journal of Surgery ; (12): 488-491, 2010.
Article in Chinese | WPRIM | ID: wpr-360755

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the clinicopathologic features, diagnosis and treatment of biliary intraductal papillary neoplasm (IPN-B).</p><p><b>METHODS</b>The clinical, histopathological, treatment and prognosis data of 23 patients with IPN-B treated from January 1998 to December 2007 were retrospectively analyzed.</p><p><b>RESULTS</b>There were 13 male and 10 female, aged from 30 to 80 years [mean age was (61 +/- 12) years]. The clinical manifestation included 10 cases with asymptomatic, 7 cases with abdominal pain, 4 cases with jaundice, 1 case with emaciation, and 1 case with acute cholangitis, respectively. Nine patients were also associated with hepatolithiasis. The average diameter of the tumors was (6 +/- 4) cm, 4 lesions were located in the right lobe, 15 in the left lobe, and 4 in the extrahepatic bile duct. Histopathologically, there were 4 adenomas, 1 borderline neoplasm, 6 carcinomas in situ, and 12 carcinomas. All patients received operation;the mean duration of follow-up was (33 +/- 28) months. Overall 3-year and 5-year survival rates of IPN-B were 85.3% and 68.2% respectively.</p><p><b>CONCLUSIONS</b>IPN-B represents a distinct clinicopathologic entity. Favorable prognosis for IPN-B is offered by curative resection.</p>


Subject(s)
Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Bile Duct Neoplasms , Pathology , General Surgery , Carcinoma, Papillary , Pathology , General Surgery , Follow-Up Studies , Prognosis , Retrospective Studies
13.
Chinese Journal of Surgery ; (12): 1536-1539, 2009.
Article in Chinese | WPRIM | ID: wpr-299678

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the clinicopathologic features, diagnosis and treatment of hepatic angiomyolipoma (HAML).</p><p><b>METHODS</b>The clinical, histopathological, treatment and prognosis data of 51 patients treated for HAML from October 1998 to October 2007 were retrospectively analyzed.</p><p><b>RESULTS</b>HAML had a female predilection (female/male = 41/10) and the mean age was 44 years old. The main symptoms were abdominal mass (33 cases) and abdominal pain or discomfort (15 cases), the other 2 cases presented as fever. Histopathologically, HAML was composed of a heterogeneous mixture of blood vessels, smooth muscle, and adipose cells. Immunohistochemical staining showed relatively high positive rate of HMB-45 (50/51), SMA (47/49) and S-100 (39/42). All 51 patients underwent partial hepatectomy. The mean hospital stay was 13.8 days and mean intraoperative blood loss was 263 ml. There was no recurrence or metastasis after a mean follow-up of 55.4 months.</p><p><b>CONCLUSIONS</b>HAML is a rare benign mesenchymal tumor of the liver. Definitive diagnosis of HAML depends on the pathohistological findings and HMB-45 positive myoid cell is an important diagnostic marker. Complete surgical resection is the optimal treatment for HAML with favorable prognosis.</p>


Subject(s)
Adolescent , Adult , Female , Humans , Male , Middle Aged , Young Adult , Angiomyolipoma , Diagnosis , Pathology , General Surgery , Follow-Up Studies , Liver Neoplasms , Diagnosis , Pathology , General Surgery , Prognosis , Retrospective Studies , Treatment Outcome
14.
Chinese Journal of Oncology ; (12): 241-245, 2009.
Article in Chinese | WPRIM | ID: wpr-293141

ABSTRACT

<p><b>OBJECTIVE</b>To evaluate the effects of specific peptide (AWYPLPP peptide) binding to high metastatic potential human hepatocellular carcinoma (HCC) cells on the invasion and metastasis of liver cancer.</p><p><b>METHODS</b>The effects of AWYPLPP peptide on the invasion, migration, proliferation and adhesion of high metastatic potential human HCC cell line (HCCLM3) were evaluated in vitro by Matrigel invasion assay, migration assay, MTT assay and adhesion assay. The effect of AWYPLPP peptide on lung metastasis of HCC in vivo was evaluated in male nude mice with subcutaneously implanted HCCLM3 cells.</p><p><b>RESULTS</b>Incubation with the AWYPLPP peptide, but not the control peptide, resulted in a concentration-dependent increase of invasion ability in HCCLM3 cells at the concentration of 0.1 to 100 micromol/L. At any concentration used for the invasion assay, the peptide had no effect on cell migration, proliferation and adhesion. After 30 days of transplantation, eight of nine (88.9%) mice in the AWYPLPP peptide group showed obvious lung metastasis. The metastatic rate of lung metastasis was significantly increased in the AWYPLPP peptide group compared with that in the control group. There was no significant difference among the weights of primary tumor in the PBS, control peptide and AWYPLPP peptide groups.</p><p><b>CONCLUSION</b>AWYPLPP peptide can promote in vitro invasion and in vivo lung metastasis of high metastatic potential human HCC cells. Identification of the receptor for AWYPLPP peptide binding may provide new insights into the molecular mechanism underlying HCC invasion and metastasis as well as new targets for intervention.</p>


Subject(s)
Animals , Humans , Male , Mice , Carcinoma, Hepatocellular , Metabolism , Pathology , Cell Adhesion , Cell Line, Tumor , Cell Movement , Cell Proliferation , Liver Neoplasms , Metabolism , Pathology , Lung Neoplasms , Mice, Inbred BALB C , Mice, Nude , Neoplasm Invasiveness , Neoplasm Metastasis , Neoplasm Transplantation , Oligopeptides , Metabolism , Pharmacology , Random Allocation
15.
Chinese Journal of Oncology ; (12): 444-447, 2008.
Article in Chinese | WPRIM | ID: wpr-357403

ABSTRACT

<p><b>OBJECTIVE</b>To examine how the thymidine phosphorylase (TP) gene expression is upregulated by interferon-alpha (IFN-alpha) in human hepatocellular carcinoma SMMC-7721 cells.</p><p><b>METHODS</b>TP mRNA levels were determined by RT-PCR. Whether the JAK-STAT cascade mediates IFN-alpha-induced TP mRNA expression was studied by pretreatment with Janus Kinase (JAK) inhibitor, AG-490. Effects of IFN-alpha on TP mRNA stability were detected with additional actinomycin D.</p><p><b>RESULTS</b>The expression of TP mRNA was induced by IFN-alpha in a dose- and time-dependent manner in SMMC-7721 (human hepatocellular carcinoma) cells. TP mRNA levels rose at 8 h, reached the peak value at 12 h, and remained at a high level up to 72 h in SMMC-7721 cells treated with IFN-alpha 10000 U/ml. IFN-alpha at a dose of 5000 or 10000 U/ml up-regulated TP expression about 3 fold compared with that of non-treated cells (P < 0.05). Induction of TP mRNA expression by IFN-alpha was significantly inhibited in SMMC-7721 cells by pretreatment with AG-490, in comparison with that treated with IFN-alpha alone. Pretreatment of SMMC-7721 cells with IFN-alpha 10000 U/ml for 24 h caused a substantial stabilization of TP mRNA, with a half-live of 35.8 h, compared with 8.5 hr in the control SMMC-7721 cells.</p><p><b>CONCLUSION</b>IFN-alpha at certain doses upregulates TP mRNA expression via both JAK-STAT transcriptional activation and post-transcriptional mRNA stabilization in human hepatocellular carcinoma SMMC-7721 cells.</p>


Subject(s)
Humans , Carcinoma, Hepatocellular , Pathology , Cell Line, Tumor , Dose-Response Relationship, Drug , Enzyme Inhibitors , Pharmacology , Gene Expression Regulation, Neoplastic , Interferon-alpha , Pharmacology , Janus Kinases , Metabolism , Liver Neoplasms , Pathology , RNA, Messenger , Metabolism , STAT1 Transcription Factor , Metabolism , Thymidine Phosphorylase , Genetics , Transcriptional Activation , Tyrphostins , Pharmacology
16.
Chinese Journal of Hepatology ; (12): 514-518, 2008.
Article in Chinese | WPRIM | ID: wpr-279755

ABSTRACT

<p><b>OBJECTIVE</b>To explore the relationships between the expression of transgelin in dendritic cells (DCs) pulsed with hepatocellular carcinoma lysates and the functions of the DCs.</p><p><b>METHODS</b>DCs derived from healthy human white blood cells were divided into 3 groups: one was pulsed with high metastatic potential hepatocellular carcinoma cell line (MHCC97H) lysates, one with lysates of a low metastatic potential cell line (MHCC97L), and one un-pulsed DCs served as the control. The morphology of the DCs was observed by confocal microscopy and scanning electron microscopy. The phenotypes of the DCs were detected by flowcytometric analysis. The mixed leucocyte reaction (MLR) test and IL-12 secretion of DCs in the supernatants of MLR were employed to determine the functions of the DCs; the expression of transgelin was detected by Western blot.</p><p><b>RESULTS</b>There were no morphological changes in the different DCs, but the levels of HLA-DR, CD80, CD83, CD86, MLR and IL-12 and transgelin were significantly higher in the two pulsed groups than those in the control group (P less than 0.01). In MHCC97H pulsed DCs, their CD80, CD83, CD86, and the expression of transgelin were also higher than those in the control group (P less than 0.05). The expression of transgelin was significantly higher in the MHCC97H pulsed group than in the MHCC97L loaded group, but CD80, CD83, CD86 and the level of IL-12 were all lower in the MHCC97H loaded DC group in comparison with those in the MHCC97 pulsed group (P less than 0.05).</p><p><b>CONCLUSION</b>The expression of transgelin in DCs pulsed with HCC lysates is related to the functions of the DCs.</p>


Subject(s)
Humans , Carcinoma, Hepatocellular , Metabolism , Cell Line, Tumor , Dendritic Cells , Metabolism , Liver Neoplasms , Metabolism , Microfilament Proteins , Muscle Proteins
17.
Chinese Journal of Hepatology ; (12): 12-16, 2008.
Article in Chinese | WPRIM | ID: wpr-277620

ABSTRACT

<p><b>OBJECTIVE</b>We previously showed that introduction of a normal, neomycin-tagged human chromosome 8 reduced the metastatic capacity of C5F rat liver cancer cell line, which had high metastatic potential without affecting tumorigenicity, suggesting the presence of one or more metastasis suppressor genes encoded on human chromosome 8. We proceeded to define further the region harboring the metastasis suppressor gene(s) and to determine the random loss of human chromosome 8 by PCR amplification of sequence tag site (STS) markers.</p><p><b>METHODS</b>The national Center for Biotechnology Information (NCBI) databases were used as references of the relative genetic distances of the STS markers. C5F genomic DNA and A9/neo8 genomic DNA were used as negative and positive controls for chromosome 8 amplification, respectively. Genomic DNA was isolated and quantified from cultured hybrid clones (A9/C5F-1 and A9/C5F-2 microcell hybrid clones served as metastasis-unsuppressed groups; A9/C5F-4, A9/C5F-8 and A9/C5F-10 microcell hybrid clones served as metastasis suppressed groups). STS-PCR products were separated by electrophoresis through 2% agarose gel.</p><p><b>RESULTS</b>Metastasis-suppressed microcell hybrid clones (A9/C5F-4, A9/C5F-8 and A9/C5F-10) conserved STS markers between D8S542 --> D8S1973 (8p21.1-23.1). In contrast, metastasis-unsuppressed clones (A9/C5F-1 and A9/C5F-2) lacked several markers in this region. In attempts to refine the region retained in the microcell suppressed clones, more densely spaced STS markers in the human chromosome 8p21.1-23.1 were used. We found that the metastasis-suppressed clones retained 18cM region between D8S542 and D8S1973 (8P21.1-23.1), where as the metastasis-unsuppressed clones lacked the region.</p><p><b>CONCLUSION</b>Our results suggest that a metastasis suppressor gene is located within the interval between D8S542 and D8S1973 on human chromosome 8p21.1-23.1.</p>


Subject(s)
Humans , Carcinoma, Hepatocellular , Genetics , Cell Line , Cell Line, Tumor , Chromosome Mapping , Chromosomes, Human, Pair 8 , Genetics , Fibroblasts , Cell Biology , Genes, Tumor Suppressor , In Situ Hybridization, Fluorescence , Liver Neoplasms , Genetics , Neoplasm Metastasis , Sequence Tagged Sites
18.
Chinese Journal of Surgery ; (12): 1621-1623, 2008.
Article in Chinese | WPRIM | ID: wpr-275966

ABSTRACT

<p><b>OBJECTIVE</b>To assess the feasibility, safety and outcome of anatomical laparoscopic left lateral hepatic lobectomy for benign and malignant liver tumors.</p><p><b>METHODS</b>From April 2005 to May 2008, 11 patients (7 male, 4 female; mean age 51.7 years) underwent anatomical laparoscopic left lateral hepatic lobectomy. Four patients presented with hepatocellular carcinoma and cirrhosis, while 1 patient had metastatic liver tumors from postoperatively colon cancer, five patients had hemangioma (2 cases with gallstones underwent cholecystectomy), 1 patient had a huge symptomatic angiolipoleiomyoma. Mean tumor size was 5.8 cm (range 2.1 to 12.0 cm). All the lesions were localized in the anatomical left lateral lobe (segments II to III).</p><p><b>RESULTS</b>The mean operative time was 147 min (range 120 to 180 min). There were no intraoperative or postoperative complications, and blood transfusions were not required. The mean postoperative hospital stay was 5.9 days.</p><p><b>CONCLUSIONS</b>Anatomical laparoscopic left lateral hepatic lobectomy are feasible and safety.</p>


Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Hepatectomy , Methods , Laparoscopy , Liver Neoplasms , Pathology , General Surgery , Treatment Outcome
19.
Chinese Journal of Hepatology ; (12): 261-265, 2007.
Article in Chinese | WPRIM | ID: wpr-230627

ABSTRACT

<p><b>OBJECTIVE</b>To compare different expression profiles of all known chemokine receptors in human hepatocellular carcinoma (HCC) cell lines with different metastasis potentials.</p><p><b>METHODS</b>Eighteen pairs of chemokine receptor primers were designed using Premier software. Expression profiles of the 18 chemokine receptors on four HCC cell lines of lower to higher potentials of metastasis (SMMC-7721, MHCC97-L, MHCC97-H and HCCLM6) were analyzed by RT-PCR. Expression of CXCR4 was detected by RT-PCR.</p><p><b>RESULTS</b>Expression profiles of chemokine receptors on four HCC cell lines with different metastatic potentials had significant differences (P < 0.01), in which CCR10, CXCR4 and CXCR6 expressions decreased gradually as the metastatic potential of the cell lines increased. The expressions of CCR3, CCR4, CCR10, CCR12 and XCR1 on HCCLM6 were significantly reduced compared with SMMC-7721 (P < 0.01), whereas the expressions of CXCR1 (P = 0.006) and CXCR5 (P = 0.003) exceeded that of SMMC-7721. Except for CXCR2, CXCR6 and XCR1, most of chemokine receptors on MHCC97-H were expressed differently compared with MHCC97-L (P < 0.05), in which expressions of CCR1 (P = 0.002), CCR2 (P = 0.004) and CCR5 (P = 0.046) exceeded MHCC97-L. CXCR4 was detected only on the positive controls and SMMC-7721 when the template of total RNA was reduced one-half in RT-PCR.</p><p><b>CONCLUSION</b>Chemokine receptors are expressed very differently at mRNA level on HCC cell lines with different metastatic potentials. The different profiles of chemokine receptors in tumor microenvironment and the function of CXCR4 in HCC should be further studied. Our findings have important implications in understanding the relationship between chemokine receptors and the metastatic potential of HCC.</p>


Subject(s)
Humans , Carcinoma, Hepatocellular , Metabolism , Pathology , Cell Line, Tumor , Liver Neoplasms , Metabolism , Pathology , RNA, Messenger , Genetics , Receptors, Chemokine , Metabolism
20.
Chinese Journal of Hepatology ; (12): 498-502, 2007.
Article in Chinese | WPRIM | ID: wpr-230553

ABSTRACT

<p><b>OBJECTIVE</b>To screen low molecular weight protein biomarkers relevant to portal vein tumor thrombi (PVTT) in serum of hepatocellular carcinoma (HCC) patients.</p><p><b>METHODS</b>Serum samples were obtained from 12 healthy volunteers, 12 HCC patients without PVTT and 12 HCC patients with PVTT. Using two-dimensional gel electrophoresis (2-DE) in which the second dimension was 16% SDS-PAGE, serum protein images of the 3 groups were analyzed by ImageMaster software. The differential protein spots were further identified by MALDI-TOF MS/MS.</p><p><b>RESULTS</b>Comparing the results using 12.5% SDS-PAGE gel, there were more protein bands (between 3 x 10(3) and 20 x 10(3)) and low molecular weight (MW) protein spots (less than 20 x 10(3)) were clearly shown in the 16% SDS-PAGE gel. Fifteen differential protein spots representing 5 proteins were found in the 3 groups by inter-class comparison and they were then identified. Compared with those in the healthy group, apolipoprotein A-I, lipoprotein CIII, transthyretin and DNA topoisomerase II were all down regulated in HCC groups and haptoglobin-2 was over expressed. All 5 proteins decreased more in the PVTT group than in the non-PVTT group.</p><p><b>CONCLUSION</b>The expression of low MW serum protein obviously changes in the beginning and in the progressive stage of HCC, and differentially expressed low MW proteins might be potential biomarkers in an early prognostic prediction and surveillance in the treatment for HCC and PVTT.</p>


Subject(s)
Adult , Female , Humans , Male , Middle Aged , Blood Proteins , Carcinoma, Hepatocellular , Blood , Pathology , Electrophoresis, Gel, Two-Dimensional , Methods , Liver Neoplasms , Pathology , Neoplastic Cells, Circulating , Pathology , Portal Vein , Pathology , Proteome
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