健脾补肾中药对肿瘤成因多阶段学说中起始和启动的影响

本实验以“细胞突变试验”,“细胞介导突变试验”作为肿瘤成因多阶段学说的起始阶段的模型,观察中医中药对它们的影响。结果提示:白术,太四君汤,仙茅,仙灵脾等健脾补肾中药具有反突变作用;太四君汤,四君子汤,枸杞子等中药具有反启动作用;从而说明某些中药对肿瘤成因多阶段学说中的起始与启动阶段有明确的阻断作用。

肿瘤就是体内的痈疽

人体的肿瘤是由于瘀,形成像体表所见的痈和疽,而内攻的痈疽就是癌。

Survey of Radiosensitizing Agents (Synthesized Chemicals and Gene Therapeutic Agents) Since 2000

Radiotherapy has played an important role in treatment of tumor patientssince it appeared about 80 years ago, and has been an indispensable part of the management of about50% of tumors (especially 60% - 70% of malignant tumors). Currently, radiotherapy is used in simpleand palliative therapy, adjuvant therapy after or before surgery, simultaneous radio-chemotherapy,combined BRM (biological response modifier) therapy, ets. Radiosensitizing agents enhance theradiation effects on tumor cells so as to have better responses in radiotherapy. Tumor intrinsicradiosensitivity is affected by the hy-poxic level in solid tumor, the ability of the cells torepair the radiation-induced DNA damage, the number of cells which have a clonogenic capability toreestablish uncontrolled cell growth, the amount of dividing cells, and the distribution of cellsthroughout the cell cycle. Consequently , it is necessary and useful to add one or moreradiosensitizing agents in radiotherapy to increase the radio-sensitivity of tumor cells.

Correlation of integrin β3 mRNA and vascular endothelial growth factor protein expression profiles with the clinicopathological features and prognosis of gastric carcinoma

AIM： To investigate integrin β3 mRNA and vascular endothelial growth factor （VEGF） protein expression in gastric carcinoma, and its correlation with microvascular density, growth-pattern, invasion, metastasis and prognosis. METHODS： In situ hybridization（ISH） of integrin β3 mRNA and immunohistochemistry of VEGF and CD34 protein were performed on samples from 118 patients with gastric cancer. RESULTS： The positive rate of integrin β3 mRNA in non-tumor gastric mucosa （20%） was significantly lower than that of the gastric cancer tissue （52.5%, X^2 = 10.20, P 〈 0.01）. In patients of infiltrating type, stage T3-T4, vessel invasion, lymphatic metastasis, hepatic or peritoneal metastasis, the positive expression rates of integrin β3 mRNA were significantly higher than those in patients of expanding type （P 〈 0.01）, stage T1-T2 （P 〈 0.01）, non-vessel invasion （P 〈 0.01）, without lymphatic metastasis （P 〈 0.01）, without hepatic and peritoneal metastasis （P 〈 0.01）, respectively. In patients of infiltrating type, stage T3-T4, vessel invasion, lymphatic metastasis, hepatic or peritoneal metastasis, the positive expression rates of VEGF protein were significantly higher than those in patients of expanding type （P 〈 0.01）, stage T1-T2 （P 〈 0.01）, non-vessel invasion （P 〈 0.01）, without lymphatic metastasis （P 〈 0.01）, without hepatic and peritoneal metastasis （P 〈 0.01）, respectively. In patients of infiltrating type, stage T3-T4, vessel invasion, lymphatic metastasis, hepatic or peritoneal metastasis, the mean MVD were significantly higher than those in patients of expanding type （P 〈 0.01）, stage T1-T2 （P 〈 0.01）, non-vessel invasion （P 〈 0.01）, without lymphatic metastasis （P 〈 0.01）, without hepatic and peritoneal metastasis （P 〈 0.01）, respectively. It was found that the positive expression rate of integrin β3 mRNA was positively related to that of VEGF protein （P 〈 0.01） and MVD （P 〈 0.05）, meanwhile the positive expression rate of VEGF protein was positively related to MVD （P 〈 0.05）. The mean survival period in patients with positive expression of integrin β3 mRNA and VEGF, and MVD ≥54.9/mm^2 was significantly shorter than that in patients with negative expression of integrin β3 mRNA （P 〈 0.05） and VEGF （P 〈 0.01）, and MVD 〈 54.9/mm^2 （P 〈 0.01）. Five-year survival rate in patients with positive expression of integrin β3 mRNA and VEGF, and MVD ≥54.9/mm^2 was significantly lower than those with negative expression of integrin β3 mRNA （P 〈 0.05）, VEGF （P 〈 0.05）, and MVD 〈 54.9/mm^2 （P 〈 0.01）. CONCLUSION： Integrin β3 and VEGF expression can synergistically enhance tumor angiogenesis, and may play a crucial role in invasion and metastasis of gastric carcinoma. Therefore, they may be prognostic biomarkers and novel molecular therapeutic targets.

A novel gain of function mutant in C-kit gene and its tumorigenesis in nude mice

AIM： To transfect mutant C-kit cDNA at codon 579 into human embryonic kidney cell line to observe its role in the pathogenesis of gastrointestinal stromal tumor （GIST）. METHODS： Eukaryotic expression vectors of pcDNA3- Kit-NW and pcDNA3-Kit-W were constructed. Then pcDNA3-Kit-NW and pcDNA3-Kit-W plasrnids were transfected into human embryonic kidney cell line by Upofectamine. The resistant clone was screened by G418 filtration and identified by sequencing, Western blotting, and immunocytochemical staining. Human embryonic kidney cells were divided into three groups including pcDNA3-Kit-NW, pcDNA3-Kit-W, and vector control groups. Absorbency value with a wavelength of 574 nm was detected by MTT analysis. Mice were injected with three groups of cells. Volume, mass, and histological examinations of the tumors in different groups were measured and compared. RESULTS： The C-kit gene and mutant C-kit gene were successfully cloned into the eukaryotic expression vector pcDNA3, pcDNA3-Kit-NW and pcDNA3-Kit-W were successfully transfected into human embryonic kidney cell line and showed stable expression in this cell line. Cell proliferating activity had significant differences between pcDNA3-Kit-NW and pcDNA3, pcDNA3-Kit- NW and pcDNA3-Kit-W （P〈0.05）, respectively. Tumors were only observed in nude mice implanted with cells transfected with pcDNA3-Kit-NW. CONCLUSION： Mutation of C-kit gene increases the proliferation activity of human cells and plays an important role in the malignant transformation of GIST.

Gene expression profiling:Canonical molecular changes and clinicopathological features in sporadic colorectal cancers

AIM： To investigate alternative or subordinate pathways involved in colorectal tumorigenesis and tumor growth, possibly determining at-risk populations and predicting responses to treatment. METHODS： Using microarray gene-expression analysis, we analyzed patterns of gene expression relative to canonical molecular changes and clinicopathological features in 84 sporadic colorectal cancer patients, standardized by tumor location. Subsets of differentially expressed genes were confirmed by real-time reverse-transcript polymerase chain reaction （RT-PCR）. RESULTS： The largest number of genes identified as being differentially expressed was by tumor location, and the next largest number by lymphovascular or neural invasion of tumor cells and by mismatch repair （NMR） defects. Amongst biological processes, the immune response was significantly implicated in entire molecular changes observed during colorectal tumorigenesis （P 〈 0.001）. Amongst 47 differentially expressed genes, seven （PISD, NIBP, BAI2, STOML1, MRPL21, MRPL16, and MKKS） were newly found to correlate with tumorigenesis and tumor growth. Most location-associated molecular changes had distinct effects on gene expression, but the effects of the latter were sometimes contradictory. CONCLUSION： We show that several differentially expressed genes were associated with canonical molecular changes in sporadic colorectal cancers, possibly constituting alternative or subordinate pathways of tumorigenesis. As tumor location was the dominant factor influencing differential gene expression, location-specific analysis may identify location-associated pathways and enhance the accuracy of class prediction.

Effects of thalidomide on angiogenesis and tumor growth and metastasis of human hepatocellular carcinoma in nude mice

AIM: To investigate the effects of thalidomide on angiogenesis, tumor growth and metastasis of hepatocellular carcinoma in nude mice.METHODS: Twenty-four nude mice were randomly divided into therapy group and control group, 12 mice in each group. Thalidomide dissolved in 0.5% sodium carboxyl methyl cellulose (CMC) suspension was administered intraperitoneally once a day at the dose of 200 mg/kg in therapy group, and an equivalent volume of 0.5% CMC in control group. Mice were sacrificed on the 30th d, tumor size and weight and metastases in liver and lungs were measured. CD34 and VEGF mRNA in tumor tissue were detected by immunohistochemistry and semi-quantitative RT-PCR respectively and microvessel density (MVD) was counted. Serum concentrations of TNF-α and ALT and AFP were also tested.RESULTS: MVD and VEGF mRNA in therapy group were less than those in control group (31.08±16.23 vessels/HP vs 80.00±26.27 vessels/HP, 0.0538±0.0165 vs 0.7373±0.1297,respectively, P＜0.05). No statistical difference was observed in tumor size and weight and metastases in liver and lungs.TNF-α was significantly lower in therapy group than in control group (28.64±4.64 ng/L vs42.69±6.99 ng/L, P＜0.05). No statistical difference in ALT and AFP was observed between groups.CONCLUSION: Thalidomide can significantly inhibitangiogenesis and metastasis of hepatocellular carcinoma.Italso has inhibitory effects on circulating TNF-α.

A fusion protein containing murine vascular endothelial growth factor and tissue factor induces thrombogenesis and suppression of tumor growth in a colon carcinoma model

Induction of tumor vasculature occlusion by targeting a thrombogen to newly formed blood vessels in tumor tissues represents an intriguing approach to the eradication of primary solid tumors. In the current study, we construct and express a fusion protein containing vascular endothelial growth factor (VEGF) and tissue factor (TF) to explore whether this fusion protein has the capability of inhibiting tumor growth in a colon carcinoma model. The murine cDNA of VEGF A and TF were amplified by reverse transcriptase polymerase chain reaction (RT-PCR), and then cloned into prokaryotic expression plasmid pQE30 with a linker. The expression product recombinant VEGF-TF (rVEGF-TF) was purified and proved to have comparable enzyme activity to a commercial TF and the capability of specific binding to tumor vessels. Significant decrease of tumor growth was found in the mice administered with rVEGF-TF on Day 6 after initiated rVEGF-TF treatment (P<0.05), and the tumor masses in 2 of 10 mice were almost disappeared on Day 14 after the first treatment. In addition, valid thrombogenesis and tumor necrosis were observed in the tumor tissues injected with rVEGF-TF. Our results demonstrate that occlusion of tumor vasculature with rVEGF-TF is potentially an effective approach for cancer therapy.

Increased expression of angiogenin in gastric carcinoma in correlation with tumor angiogenesis and proliferation

AIM： To investigate the implication of angiogenin （ANG） in the neovascularizaton and growth of human gastric carcinoma （HGC）. METHODS： ANG mRNA expression in HGC specimens obtained by surgical resection from patients with HGC were examined by RT-PCR. ANG, Ki-67, VEGF protein expression and microvessel density （MVD） in HGC specimens were detected by immunohistochemistry. RESULTS： RT-PCR showed significantly higher ANG mRNA expression （0.482 ± 0.094） in HGC tissues than in the surrounding nontumorous tissues （0.276 ±0.019, P = 0.03）. MVD within tumorous tissues increased significantly with ANG mRNA expression （r = 0.380, P = 0.001） and ANG protein expression （P 〈 0.01）. The ANG expression levels of cancer tissues were positively correlated with VEGF （P 〈 0.01） and the proliferation index of cancer cells （P 〈 0.01）. CONCLUSION： ANG is one of the neovascularization factors of HGC. ANG may work in coordination with VEGF, and promote the proliferation of HGC cells.

Anti-angiogenesis in hepatocellular carcinoma treatment: Current evidence and future perspectives

Hepatocellular carcinoma(HCC) is among the most common cancer diseases worldwide.Arterial hypervascularisation is an essential step for HCC tumorigenesis and can be targeted by transarterial chemoembolization(TACE).This interventional method is the standard treatment for patients with intermediate stage HCC,but is also applied as "bridging" therapy for patients awaiting liver transplantation in many centers worldwide.Usually the devascularization effect induced by TACE is transient,consequently resulting in repeated cycles of TACE every 4-8 wk.Despite documented survival benefits,TACE can also induce the up-regulation of proangiogenic and growth factors,which might contribute to accelerated progression in patients with incomplete response.In 2007,sorafenib,a multi-tyrosine kinase and angiogenesis inhibitor,was approved as the first systemic treatment for advanced stage HCC.Other active targeted compounds,either inhibitors of angiogenesis and/or growth factors,are currently being investigated in numerous clinical trials.To overcome revascularisation or tumor progression under TACE treatment it seems therefore attractive to combine TACE with systemic targeted agents,which might theoretically block the effects of proangiogenic and growth factors.Over the last 12 mo,several retrospec-tive or prospective cohort studies combining TACE and sorafenib have been published.Nevertheless,robust results of the efficacy and tolerability of such combination strategies as proven by randomized,controlled trials are awaited in the next two years.

Characterization of FGFR signaling pathway as therapeutic targets for sarcoma patients

The fibroblast growth factor receptor(FGFR) family plays important roles in regulating cell growth, proliferation, survival,differentiation and angiogenesis. Deregulation of the FGF/FGFR signaling pathway has been associated with multiple development syndromes and cancers, and thus therapeutic strategies targeting FGFs and FGFR in human cancer are currently being explored.However, few studies on the FGF/FGFR pathway have been conducted in sarcoma, which has a poor outcome with traditional treatments such as surgery, chemotherapy, and radiotherapy. Hence, in the present review, we provide an overview of the role of the FGF/FGFR pathway signal in sarcoma and FGFR inhibitors, which might be new targets for the treatment of sarcomas according to recent research.

Expressions of MVD, VEGF, Ki67 in Residual Prostate Cancer after Cryoablation

OBJECTIVE To analyze the effects of cryoablation on the mice bearing Rm-I prostate cancer through detecting tumor angiogenesis and cancer cell proliferation in the mice after cryoablation, and to explore the effects of cryoablation on vascular endothelium growth factor （VEGF）, Ki67 protein expression and microvessel density （MVD） in the mice bearing prostate cancer. METHODS Sixty Rm-1 mouse models of prostate cancer were established. Experimental mice were randomized into 2 groups： the cryoablation group （n = 30） and the control group （n = 30）. After file therap）4 tumor tissues of the mice in group A and B were obtained at day 0 （without cryoablation）, 1st, 3rd, 5th, 7th, 14th day, respectivelj6 after cryoablation, and the expressions of MVD, VEGF and Ki67 proteins were detected at the same time points. RESULTS The expressions of MVD, VEGF and Ki67 proteins in group A were decreased. The lowest values of the factors were detected on the 3rd day after cryoablation, and increased slowly after that. The expressions of MVD, VEGF and Ki67 proteins in the control group were not changed. Significant changes of the expressions of MVD, VEGF and Ki67 proteins in the group A were found at different time points. Correlation analysis suggested a positive correlation between the expressions of VEGF and MVD proteins （r = 0.8793）, a positive correlation between the expressions of Ki67 and MVD proteins （r = 0.7614）, and a positive correlation between the expressions of VEGF and ki67 proteins （r = 0.6921）. CONCLUSION After argon-helium cryoablation treatment for the mice bearing prostate cancer, the expressions of MVD, VEGF and Ki67 proteins in local tumor were reduced on the 1st day. The lowest values of the factors were detected on the 3rd day after cryoablation, and then increased after that. Cryoablation combined with other modalities of treatment may effectively improve the treatment effects of cryoablation for prostate cancer.

Correlation of VEGF expression,angiogenesis and carcinomatous change in breast tumors

Objective: The aim of the study was to detect the expression of vascular endothelial growth factor (VEGF) and microvessel density (MVD) in breast benign tissues and malignant tumors to clarify the relationship between VEGF expression, angiogenesis and breast carcinoma occurrence. Methods: The expression of VEGF and MVD in 79 cases of invasive ductal breast carcinoma, 79 corresponding para-cancer normal tissues from primary invasive breast carcinomas, 35 breast carcinoma in situ, 23 breast atypical hyperplasia and 56 breast fibroid tumor were examined by immunohistochemistry staining (SP-method). Results: The positive rate of VEGF and MVD value increased significantly with the increase of the malignant degree of breast tissues (P = 0.000). In breast carcinoma group, the positive rate of VEGF and MVD value with lymph node metastasis were higher than those without lymph node metastasis (P = 0.011 and P = 0.023). A significant higher expression of VEGF and MVD value were observed as the clinical stage increased (P = 0.035 and P = 0.012). The MVD value was higher in VEGF positive group than negative group (P = 0.000). Conclusion: The combining detection of VEGF expression and MVD is helpful for evaluating malignant degree of breast carcinoma. Angiogenesis in breast tumors and occurrence of breast carcinoma might be correlated with the expression of VEGF.

The effect of rh-endostatin on micrangium and angiogenic factors in tumor and myocardium tissue

Objective： The aim of this study was to compare effect of rh-endostatin on microvasculature in tumor and myocardium tissue. Methods： Nude mice were randomized into 4 groups, blank control group [did not burden tumor, normalsaline （NS） 100 μL/d], drug control group （did not burden tumor, rh-endostatin 400 μg/d）, model group （mice burdened tumor, NS 100 μL/d） and treatment group （mice burdened tumor, rh-endostatin 400 μg/d）, administration was given during d1-d28. The volume of tumor and the weight of mouse were measured before and after administration. The expression of CD34, MMP-2, MMP-9, HIF-la and VEGF in myocardium and tumor were detected by immunohistochemistry. The structure of vasculature was observed by immunoenzymatic double staining with CD34 and Masson. Results： The tumor volume increase of treatment group （48.18 mm3） was less than the model group （113.80 mm3）, the change of weight was not significant among the four groups. After treated with endotar, the expression of MMP-9 and VEGF in tumor were obviously down-regulated, but the same results was not found in MMP-2, HIF-la of tumor. MVD in tumor of treatment group decreased significantly compared with model group. Proportion of tumor vessels covered by collagen in treatment group increased compared with model group. However, MVD and microvasculature in myocardium did not change significantly. Conclusion： Rh-endostatin can decrease the expression of MMP-9, VEGF and MVD to inhibit growth of tumor and normalize micrangium in tumor but cannot weaken MMPs and MVD of mature micrangium in myocardium.

Exploring the phytoconstituents targeting TNF-αas potential lead compounds to treat inflammatory diseases:an in-silico approach

Objective To explore the anti-inflammatory phytoconstituents from various plant sources as tumour necrosis factor-α(TNF-α)-inhibitor,a mediator involved in the inflammatory disorder,by in silico molecular docking.Methods Based on previous findings,we performed the in silico assessment of anti-inflammatory phytoconstituents from different medicinal plants to understand their binding patterns against TNF-α(PDB ID:6OP0)using AutoDock Vina.Molecular docking was performed by setting a grid box(25×25×25)Åcentered at[-12.817×(-1.618)×19.009]A with 0.375A of grid spacing.Furthermore,Discovery Studio Client 2020 program was utilized to assess two-and three-dimensional(2D and 3D)hydrogen-bond interactions concerning an amino acid of target and ligand.Physicochemical properties were reported using the Lipinski’s rule and SwissADME database to support the in silico findings.Results From the selected medicinal plants,more than 200 phytocompounds were screened against TNF-α protein with binding scores in the range of -12.3 to -2.5 kcal/mol.Amongst them,emodin,aloe-emodin,pongamol,purpuritenin,semiglabrin,ellagic acid,imperatorin,α-tocopherol,and octanorcucurbitacin A showed good binding affinity as -10.6,-10.0,-10.5,-10.1,-11.2,-10.3,-10.1,-10.1,and -10.0 kcal/mol,respectively.Also,the absorption,distribution,metabolism,excretion,and toxicology(ADMET)profiles were well within acceptable limits.Conclusion Based on our preliminary findings,we conclude that the selected phytoconstituents have the potential to be good anti-inflammatory candidates by inhibiting the TNF-α target.These compounds can be further optimized and validated as new therapeutic components to develop more effective and safe anti-inflammatory drugs.