Ubiquitin-like proteins are structurally similar to ubiquitin. These proteins are processed, activated, conjugated, and re-leased from conjugates by enzymatic steps that are similar to the corresponding mechanisms for ubiquitin. Ubiquitin-like proteins regu-late a wide array of cellular processes through modification processes, such as nuclear-cytosolic transport, transcriptional regulation, protein stability, response to stress, and progression through the cell cycle. A large number of recent studies have found dysfunctional ubiquitin-like proteins in hepatocellular carcinoma. These proteins are important in tumorigenesis, cell proliferation, apoptosis, and an-giogenesis. Anticancer drug studies revealed that regulating protein modification by using ubiquitin-like proteins may alter the anti-tu-mor effects of chemotherapy and thus influence the chemosensitivity of hepatocellular carcinoma. Results indicate that ubiquitin-like proteins may become a new target for cancer therapy. The mechanism of ubiquitin-like proteins in tumorigenesis and hepatocellular car-cinoma progression is of great significance in the diagnosis and treatment of hepatocellular carcinoma.

Objective To set up an apoptosis model of prostate cancer cell line and to clone and study the apoptosis related genes. Methods An apoptosis model of prostate cancer cell line DU-145 has been set up through induction by all transretinoic acid (ATRA).During the process of cell apoptosis the apoptosis-related gene was cloned by means of improved PCR-based subtractive hybridization from the apoptosis prostate cancer cell line DU-145 prostate cancer cells. Results During the process of DU-145 cell apoptosis,c-erb B-2 expression,TNF genes and some unknown apoptosis-related gene were observed.This has been accepted by Genebank,the accession number being AF174394. Conclusions ATRA-induced apoptosis of DU-145 cells is a complex process with multiple genes involved,some of which being unknown yet.

Objective To develop a new method to repair the defect after enlarged excision of dermatofibrosarcoma protuberans.Methods This study included 8 patients with dermatofibrosarcoma protuberans measuring 2.5 to 5.5 cm in diameter.All the patients underwent enlarged excision of the affected skin and subcutaneous tissue.The defect measured 12.5 to 17.5 cm in diameter.Sieve skin flaps secured with a vacuum-assisted closure device were used to repair the huge surface defects.Results All the patients experienced the survival of sieve skin flaps at stage Ⅰ after operation,with no infection,effusion or necrosis.No relapse was observed during the 3 to 40 months of follow up.A satisfactory recovery was achieved in skin appearance and function with the formation of a flat scar,and no obvious proliferation occurred.Conclusion The vacuum-assisted closure device offers a safe and simple method for securing skin grafts to the defect after enlarged excision of dermatofibrosarcoma protuberans.

Objective To predict as well as bioinforrmatically analyze the target genes of has-miR-451.Methods miRBase, miRanda, TargetScan and PicTar were used to predict the target genes of hsa-miRNA-451.The functions of the target genes were demonstrated by Gene Ontology and pathway enrichment analysis.P＜0.05 was set as statistically significant.Results 18 target spots of hsa-miRNA-451 were predicted by 3 databases or prediction software at least.The functions of the target genes were enriched in proliferation and development of epithelial cells and regulation of kinase activity (P＜ 0.05).Pathway analysis showed that transforming growth factor-beta signaling pathway, mitogen-activated protein kinase signaling pathway, epidermal growth factor signaling pathway, Wnt signaling pathway and mammalian target of rapamycin signaling pathway were significantly enriched (P＜0.05).Conclusion hsa-miRNA-451 might be involved in various signaling pathways related to proliferation and development of epithelial cells.

Objective To observe the level of the nitric oxide (NO) and endothelin-1 (ET-1) in the exhaled breath condensate(EBC)and serum of the patients with ALI /ARDS, and investigate its clinical significance. Methods The study group included 52 mechanical ventilation patients with ALI/ARDS in ICU , which were divided into the survival and death group, while 30 healthy volunteers were recruited as healthy control. EBC samples of the healthy control and the study group on the 1st day and 5st day were collected by EcoScreen condenser with the synchronous collection of the venous blood. The concentrations of NO and ET-1 in the EBC and serum were measured by EIA. Results The levels of NO and ET-1 in EBC and serum of the patients with ALI /ARDS were all significantly higher than those of the healthy control. After treatment , the levels of NO and ET-1 in EBC and serum of the patients all decreased significantly compared with before treatment. After treatment , The levels of NO in EBC and serum of the survival group were significantly lower than those of the death group. After treatment , the levels of ET-1 in serum of the survival group was significantly lower than that of the death group. Conclusions Detecting the levels of NO and ET-1 in the EBC and serum can reflect oxidative stress , inflammatory reaction and endothelial injury in lung of patients with ALI/ARDS.

OBJECTIVE:To investigate clinical efficacy of oxaliplatin+calcium folinate+5-fluorouracil(mFOLFOX6),oxalipl-atin+capecitabine(CapeOX),irinotecan+calcium folinate+5-fluorouracil(FOLFIRI)for metastatic colorectal cancer,and to conduct cost-effectiveness analysis. METHODS:48 patients with colorectal cancer were divided into mFOLFOX6 group(30 cases),Cape-OX group(8 cases)and FOLFIRI group(10 cases). Clinical efficacy and ADR of 3 groups were analyzed,and cost-effectiveness analysis was also conducted. RESULTS:Clinical effective rates of mFOLFOX6 group,CapeOX group and FOLFIRI group were 96.67%,87.50% and 80.00%,respectively,the mFOLFX6 group was significantly higher than the other 2 groups,with statistical significance(P<0.05). mFOLFOX6 group had high incidence of gastrointestinal side effects (70.00%). FOLFIRI group had high incidence of myelosuppression (70.00%). CapeOX and mFOLFOX6 group suffered from liver injury possibly,without statistical significance (P>0.05). The C/E of mFOLFOX6 group,CapeOX group and FOLFIRI group were 11 950,15 674 and 18 397 re-spectively,to which results of sensitivity analysis were same. CONCLUSIONS:The cost-effectiveness of mFOLFOX6 regimen is superior to CapeOX and FOLFIRI regimen in the treatment of metastatic colorectal cancer,but it has the high incidence of gastroin-testinal side effects.

OBJECTIVE: To study the effect of various polarity fractions extracted from Sancaofang (SCF) and its combination on proliferation of human lung adenocarciaoma SPC-A-1 cells for bolting the most effective position of anti-tumor and suitable compatibility regimes.METHODS: Ethanol extraction and water extraction were adopted to prepare 95%,60% and 30% ethanol extract portion,water extract portion of SCF and compound decoction.The MTT assay was used to determine the effect of various polarity fractions of SCF,decoction and its combination on SPC-A-1 cells proliferation.RESULTS: IC50 of 60% ethanol extract was the smallest for SPC-A-1 cells.60% ethanol extract combined with 95% ethanol extract acts as a stimulus to anti-tumor activity significantly.CONCLUSION: The best suitable compatibility regimes were 95% ethanol extract combined with 60% ethanol extract.The liposolubility extract of SCF can be applied for anti-tumor.

OBJECTIVE:To establish the method for the simultaneous determination of anticancer activity components of flavonoids from Hedyotis diffusa,i.e. quercetin,kaempferol. METHODS:HPLC was applied to determine the contents and performed on Alltima C18(250 mm?4.6 mm,5 ?m) column. Mobile phase consisted of methanol(A)-0.5% glacial acetic acid,(gradient elution). The detection wavelength was aet at 350 nm. RESULTS:The linear range of quercetin was 0.006 2～0.244 0 ?g(r=0.999 8)and that of kaempferol 0.007 8～0.310 6 ?g(r=0.999 9). The average recovery of quercetin was 101.84%(RSD=1.79%,n=6) and that of kaempferol 99.04%(RSD=2.90%,n=6). CONCLUSIONS:The method is simple,accurate and reproducible for the quality control of H. diffusa.

Objective To investigate the detection of early cervical cancer patient's peripheral blood CK19 mRNA and its clinical significance. Methods 30 early (ⅠA2~ⅡA) cervical cancer patients from the department of gynecological tumors in the third affiliated hospital of Kunming Medical University were selected as experimental group, 15 patients with uterine myoma and 15 healthy volunteers as negative control group,15 advanced (ⅡB~ⅢB) cervical cancer patients as positive control group. Peripheral blood samples before receiving radical resection of experimental group,and the same samples before treatment of control groups were collected. The change of relative expression level and positive rate of peripheral blood CK19 was analyzed using qRT-PCR technique, and its correlation with clinicopathological parameters and prognosis was further analyzed. Results There was significant difference in relative expression level of CK19 mRNA in the peripheral blood between experimental group and control groups ( <0.001) . And the positive rate of CK19 in the peripheral blood from 30 patients with early cervical cancer before operation in the experimental group was 30% (9/30) . Furthermore, CK19 mRNA positive rate before surgery was correlated with lymph node metastasis, muscularis invasion and pathologic risk factors for postoperative ( <0.05) . There were no significant correlation between CK19 mRNA positive rate and other clinical and pathological features such as age,clinical stage (FIGO),tumor cell differentiation,tumor size,pathological type and etc ( > 0.05) . Conclusions CK-19 mRNA is a specific and suitable molecular marker for the detection of circulating tumor cells in early cervical cancer. Detection of peripheral blood CK19 mRNA expression before receiving radical hysterectomy in patients with early cervical cancer can also make up for the disadvantage of other methods such as the imaging, tumor marker detection. Most important of all, it can provide the basis for choosing a proper adjuvant therapy post-operation and estimating the relapse and prognosis.

Aim To construct the cell model targeted on the damage by α-synuclein for screening anti-Parkinson’s Disease (PD)compounds.Methods The cDNA fragment of α-synucle-in gene was obtained by PCR methods and inserted into the re-combinant prokaryotic plasmid by molecular cloning technique. The recombinant plasmid was transformed into Escherichia coli, and subsequently induced to express α-synuclein protein.The recombinant α-synuclein was purified and identified by affinity chromatography,immunoblotting and mass spectrometry.The cells damage by α-synuclein was evaluated through cell viability measured by 3-(4,5-dimethyl-2-thiazolyl )-2,5-diphenyl-2-H-tetrazolium bromide.Results The obtained cDNA fragment ofα-synuclein in accordance with its theoretic molecular weight was cloned into pET30a plasmid and verified by sequencing.The re-combinant plasmid was transformed into bacteria E.Coli.BL21 (DE3)and induced to express α-synuclein by isopropyl β-D-1 -thiogalactopyranoside (IPTG).The expression condition was op-timized according to the culture temperature,the concentration of IPTG and the proliferation state of bacteria.The purified α-synu-clein was proved to be a 1 5.3 ku molecule weight protein,and could be immunoblotted with anti-α-synuclein antibody.The pu-rified α-synuclein could decrease the viability of PC1 2 cells and primary neurons significantly,and its effect was in a concentra-tion-dependent manner.Conclusion We have succeeded in constructing the cell model targeted on the damage by α-synucle-in.