OBJECTIVE:To investigate the in vitro transdermal absorption properties of galangin and effects of different pene-tration enhancers on its transdermal behaviors,and provide reference for developing skin preparations using galangin as APIs in the treatment of vitiligo. METHODS:HPLC was used to determine the galangin content. Using cumulative permeation rate (Q) and the transdermal rate(J)of galangin as indexes,the effect of absorption of receiving solution [20%,40% polyethylene glycol 400 (PEG400)solution and 30% ethanol solution] and rotating rate(200,300,400 r/min)on galangin in complete skin of mice were investigated,as well as the azone(1%,3%,5%)and propylene glycol(10%,20%,40%)alone or combination on its penetra-tion promotion. And the transdermal properties of galangin in complete skin,exfoliating skin,dermis skin of rats and mice were de-tected. RESULTS:The best permeability of complete skin of mice showed in 40% PEG400 solution at rotating speed of 300 r/min with 5% azone alone,J was 3.2570 μg/(cm2·h). Js of complete skin,exfoliating skin,dermis skin of mice were 2.7199,34.016, 33.874 μg/(cm2·h),respectively;and those of rats were 0.4996,9.5124,17.406 μg/(cm2·h). CONCLUSIONS:Galangin can penetrate the complete skin of mice and rats,however,the penetration quantity is far lower than exfoliating skin and dermis skin.

Objective To construct plvx-cyclooxygenase-2(COX-2)-DsRed and establish breast cancer cell line MCF7 which overexpressed COX-2, to explore the effect of COX-2 on breast cancer cell.Methods The full-length COX-2 PCR product was obtained by total COX-2 PCR primers and COX-2 cDNA vector.After the PCR product and lentiviral vector plvx-DsRed-Monomer-N1 were cut simultaneously by restriction enzyme BamH1 and Xholl, they were connected and sequenced, to get lentiviral vector plvx-COX-2-DsRed.After selected by puromycin, overexpressed COX-2 breast cancer cell line MCF7-plvx-COX-2-DsRed was obtained.The stable cell line was verified by real time PCR and Western blot.The differences of proliferation ability between stable cell line and normal one were compared by colony formation test and Western blot.Results The lentiviral vector plvx-COX-2-DsRed and stable cell line MCF7-plvx-COX-2-DsRed after selecting were obtained.COX-2 expression level of the stable cell line was 75.29 times as high as that of MCF7, and 64.91 times as high as that of cell line MCF7-plvx-DsRed-Monomer-N1 by PCR assay (P ＜ 0.05), which was consistent with the results of Western blot and microscope photo.MTT results showed that cell line MCF7-plvx-COX-2-DsRed had grown faster than cell line MCF7 and MCF7-plvx-DsRed-Monomer-N1 from the 2nd day (P ＜ 0.05), which was accordant with colony formation assay.MCF7-plvx-COX-2-DsRed cell line had higher c-myc expression and lower β-catenin expression than MCF7 cell and cell line MCF7-plvx-DsRed-Monomer-N1 detected by Western blot relative quantification (P ＜ 0.05).Conclusion The plvx-COX-2-DsRed lentiviral vector and cell line MCF7-plvx-COX-2-DsRed are successfully constructed.COX-2 can increase proliferation of MCF7 cells through up-regulating the expression of c-myc.

It is one of the main characters of malignant tumors that malignant tumor cells invade surrounding tissues and metastasize to distant tissues .Multiple factors are involved in this complicated dynamic process .Metastasis is the major factor influencing recurrence and prognosis .Therefore, it is important to explore the mechanism of invasion and metastasis for reducing recurrence rate and mortality of malignant tumors .Engulfment and cell mobility ( ELMO) family is one kind of conserved protein in evolutional process .It includes 3 members, ELMO1, ELMO2 and ELMO3.The members of ELMO family play an important role in cell phagocytosis and cell migration , and they also have close correlation with ma-lignant tumor cell invasion and metastasis .In this paper , we review the progress of the relationship between ELMO family and malignant tumor invasion and metastasis .

Objective To figure out and verify infiltration related miRNAs in bladder urothelial carcinoma (BUC). Methods Fresh tissues (20 samples,12 were infiltrative BUC samples,8 were non-infiltrative BUC samples) were collected in liquid nitrogen.The total RNA was extracted by using Trizol reagents.RNA quality control; miRNA microarray hybridization; data analysis.Another 22 samples were collected in fresh (15 were infiltrative BUC samples,7 were non-infiltrative BUC samples) for verifying purpose.4 types of bladder cancer cell lines were used for the study.BUC cell strain; total RNA was extracted by Trizol reagents; RNA quality control; RT-PCR and analysis of the data. Results ①In infiltrative BUC group,compared with non-infiltrative BUC group,there were 7 differentially expressed miRNAs:hsa-miR29c,hsa-miR-200a,hsa-miR-378,hsa-miR-429,hsa-miR-200c and hsa-miR-141 were up-regulated; hsamiR-451 was down-regulated.②In collected samples,the result of RT-PCR was consistent with miRNA array.③In bladder cancer cell lines,only the results of T24 were consistent with miRNA array. Conclusion Infiltration of BUC might relate with different expression of miRNAs.

To get high cell density,pH-stat fermentations of L.casei Zhang were carried out in the medium optimized previously under different cultural conditions.The influences of neutralizing agents,the concen-tration of buffer salts and glucose in the medium,pH,aeration and fed batch fermentation on the growth of L.casei Zhang were investigated.Based on the values of maximum specific growth rate,biomass and viable cells count in different cultural conditions,the optimal growth condition was regarded as follows:Glucose level of the medium was 80 g/L ～100 g/L;The pH was kept constant at 5.9 with aqua ammonia and anae-robic condition was kept by sparged with nitrogen periodically;The temperature was set at 37?C and cul-tured for 10 h ～12 h.The biomass and viable cells' number of L.casei Zhang under this culture conditions were 7 g/L and 3.5?1010 CFU/mL respectively,and were 7 times as high as that before optimized,which can meet the requirements of probiotics.

OBJECTIVE The aim of the present study was to evaluate the protective effects of momordica charantia polysaccharides(MCP)on depressive animal model induced by chronic social defeat stress(CSDS)and explore the underlying mechanisms.METHODS We established CSDS depressant mouse model and treated CSDS mice with MCP.Sucrose preference,forced swim test(FST)and social interaction test(SIT)were used to measure behaviors changes.We used ELISA,Q-PCR and western blot to test the levels of cytokines in the hippocampus. RESULTS The results showed that chronic administration of MCP(100,200 and 400 mg·kg-1)significantly prevented depressive-like behaviors in mice as assessed by social interaction (SIT), tail suspension (TST) and sucrose preference tests (SPT).It was showed that the elevation of proinflammatory cytokines(TNF-α,IL-6,and IL-β)concentra-tions,up-regulation of JNK3,c-Jun,and P-110β protein expressions in the hippocampus of CSDS model. Moreover,reduction activity of PI3K and phosphorylation level of protein kinase B(AKT)was also observed in the hippocampus of CSDS model.All above phenomenon were reversed after MCP intervened.Further-more,the protective effects of MCP on the CSDS mice were partly inhibited by the specific phosphati-dylinositol 3-kinase (PI3K)inhibitor, LY294002. CONCLUSION The protective effects of MCP against depressive-like effects in CSDS mice might reduce neuroinflammatory and involve in attenuation of JNK3/PI3K/AKT pathway in the hippocampus.

The effects of combined RNA interference (RNAi) of human telomerase RNA (hTR) and human telomerase reverse transcriptase (hTERT) genes on telomerase activity in a bladder cancer cell line (BIU-87 cells) were investigated by using gene chip technology in vitro with an attempt to evaluate the role of RNAi in the gene therapy of bladder transitional cell cancer (BTCC). Three TR-specific double-stranded small interfering RNAs (siRNAs) and three TERT-specific double-stranded siRNAs were designed to target different regions of TR and TERT mRNA. The phTR-siRNA, phTERT-siRNA, and the combination of both plasmids phTR+phTERT-siRNA were transfected into BIU-87 cells. The expression of hTR and hTERT mRNA was detected by quantitative fluorescent reverse transcription-polymerase chain reaction, and a telomeric repeat amplification protocol was applied to detect telomerase activity. Growth inhibition of BIU-87 cells was measured by MTT assay. Gene chip analysis was performed to evaluate the effects of the combined RNAi of hTR+hTERT genes on telomerase activity and growth of BIU-87 cells in vitro. The results showed that the expression of hTERT and hTR mRNA was inhibited by pRNAT-hTERT-III, pRNAT-hTR-III, and pRNAT-hTR-III+hTERT-III in BIU-87 cells. The inhibition efficiency of pRNAT-hTERT-III, pRNAT-hTR-III, pRNAT-hTERT-III+pRNAT-hTR-III was 67% for TERT mRNA, 41% for TR mRNA, 57% for TR mRNA and 70% for TERT mRNA in BIU-87 cells respectively. The growth of BIU-87 cells was inhibited and telomerase activity was considerably decreased, especially in the cells treated with combined RNAi-hTR and -hTERT. Gene chip analysis revealed that 21 genes were down-regulated (ATM, BAX, BCL2, BCL2L1, BIRC5, CD44, CTNNB1, E2F1, JUN, MCAM, MTA1, MYC, NFKB1, NFKBIA, NME4, PNN, PNN, SERPINE1, THBS1, TNFRSF1A, and UCC1). The results indicated that hTR-siRNA and hTERT-siRNA, especially their combination, siRNA hTR+hTERT, specifically and effectively suppressed the expression of both hTR and hTERT mRNA and telomerase activity. Molecular biological mechanism by which combined siRNA-TR and -TERT inhibited telomerase activity and growth of BIU-87 cells in vitro may involve the down-regulation of the 21 genes.

BackgroundIt has been proved that as a chemokine,interferon-inducible protein-10(IP-10)can regulate the immuno-inflammatory reaction.Some new researches showed that IP-10 also played role in regulating the neovascular vessel formation.Corneal neovascularization (CNV) is associated with multiple cellular factors,but its mechanism is below clear.Objective The present study was to address the roles of exogenous mouse IP-10 in alkali burn-induced CNV.Methods Eighty-two SPF BALB/c mice were used in this experiment and grouped according to random number table.The corneal alkali burn models were established by putting the filter paper with 1 mol/L NaOH at the central corneas of the left eyes for 40 seconds.10 mg/L IP-10 was topically administered from the first day or 14 days after modeling in the early intervene group( 10 eyes)or middle-late intervene group(5 eyes).CNV area was measured as a percentage of whole cornea.0.2％ sodium hyaluronate(HA) as vehicle was utilized in the model control group.Angiogenic factor expression in corneal tissue in the early intervene group was quantified by reverse transcriptase polymerase chain reaction (RT-PCR)and compared with model control group.All animal experiments were performed in accordance with the Association for Research in Vision and Ophthalmology Statement for the Use of Animals in Ophthalmic and Vision Research and complied with the standards of Guidelines for the Care and Use of Laboratory Animals of Soochow University. ResultsThe CNV percentage was(88.67±10.22) ％ in the model control mice,showing a significant increase in comparison with that of IP-10 early intervene group (70.06±12.21)％ (t=3.77,P=0.00).In 21 days after corneal alkali burn,the CNV percentage was(87.33±13.47)％ in the model control mice,and that of the IP-10 middle-late intervene group was ( 86.56± 12.47 ) ％ without significant difference between them ( t =1.26,P＞0.05 ).Two days or four days after IP-10 early intervene,the expressions of chemokine receptor type 3 ( CXCR3 ) in corneal tissue were significant higher than model control group( t =3.13,3.07,P＜0.05 ),but the expressions of vascular endothelial growth factor (VEGF) in cornea were lowed ( t =5.99,6.27,P＜0.01),and so were transforming growth factor-β1 (TGF-β1) (t =8.50,P＜0.01;t =4.53,P＜0.05).Conclusions The early topical administration of the exogenous mouse IP-10 can inhibit CNV by up-regulating CXCR3 expression and down-regulating VEGF and TGF-β1 expression in cornea.However,middle-later usage of the IP-10 is uneffective.

AIM:To reveal the role and function of engulfment and cell mobility 1 (ELMO1) in the invasion and migration of gastric cancer cells.METHODS:The expression of ELMO1 at protein and mRNA levels was detected in 5 kinds of gastric cancer cells and 1 normal human gastric epithelial cells by Western blot and real-time PCR, and the gastric cancer cells with the highest expression of ELMO1 were screened out.The cell transfection experiment was used to silence ELMO1 expression in the gastric cancer cells, and the effect of ELMO1 silencing on the invasion and migration of the gastric cancer cells was detected by Transwell assay.RESULTS:The expression level of ELMO1 in the gastric cancer cells was significantly higher than that in human normal gastric epithelial cells (P<0.01), and the SGC7901 cells had the highest expression level of ELMO1.ELMO1-siRNA significantly silenced the expression of ELMO1 in the SGC7901 cells (P<0.01).Silencing of ELMO1 expression significantly reduced the invasion and migration abilities of the human gastric can-cer cells (P<0.01).CONCLUSION:ELMO1 is highly expressed in the gastric cancer cells, and promotes the invasion and migration abilities of the gastric cancer cells.ELMO1 may become an effective target for the treatment of invasion and metastasis of gastric cancer.

Objective To analyse the diagnosis and treatment options of serous cystadenoma of the pancreas.Method The clinical data of 57 patients operated in the Tianjin Medical University Cancer Institute & Hospital from August 1996 to December 2011 with pathologically confirmed serous cystadenoma of pancreas after the operation were retrospectively studied.Results There were 13 males (22.8％) and 44 females (77.2％).The median age was 56.8 years.The patients were asymptomatic in 31.6％.CT was accurate in the diagnosis in 70.6％.All patients received surgical resection,inluding pancreaticoduodenectomy (n =17,29.8％),distal pancreatectomy (n =38,66.7％),palliative resection (n=1),and tumor enucleation (n=1).Postoperative complications developed in 6 patients.Histopathologically,there were 50 cases of serous microcystic adenoma (87.7％) and 7 cases of serous oligocystic adenoma (12.3 ％).One of these patients had developed into serous cystadenocarcinoma.At a follow-up of 12 months to 15 years,one patient with serous cystadenocarcinoma died 13 months after the operation.The remaining patients were all alive.Statistical analysis was performed based on the postoperative histopathological type and tumor size.The mean postoperative hospital stay of the group of patients with serous microcystic adenoma were significantly longer than the patients with serous oligocystic adenoma [(17.39±7.61) d vs (19.43±0.98) d,P=0.002].The incidence of patients with clinical symptoms was higher in the group of patients with tumor size ≥4 cm when compared with the patients with tumour size ＜4 cm.There was no significant difference on the other parameters.Conclusions Pancreatic serous cystadenoma is a rare pancreatic tumor,and it often happens in elderly women.Indications for surgical resection included symptomatic tumours,tumor diameter more than 4 cm,malignant biological behavior,malignancy could not be ruled out,and potentially malignant tumors.For asymptomatic patients and tumor size less than 4 cm,surgical resection should also be considered if the tumour progresses on follow-up.