Objective To investigate the difference between two substrains,5-8F and 6-10B,of nasopharyngeal carcinoma cell line SUNE1on expressing metastasis-associated gene 1 (MTA1),and evaluate the silencing effect of lentivirus-mediated RNA interference (RNAi) on MTA1 in human nasopharyngeal carcinoma cell line.Methods The differential expression of MTA1 mRNA and MTA1 protein in 5-8F and 6-10B cell lines were detected by real time PCR and Western blotting respectively.Short interference RNA (siRNA) fragment targeting MTA1 gene was designed using online databases and software.A specific RNAi lentiviral vector targeting human MTA1 gene was constructed and transfected into nasopharyngeal carcinoma cell line 5-8F.Real time PCR and Western blotting were performed to detect the expressive changes in MTA1 mRNA and MTA1 protein in the transfected 5-8F cells respectively.Results Compared with cell line 6-10B,the expressions of MTA1 mRNA and MTA1 protein were higher in 5-8F cell line.Sequence analysis validated the correct insert of siRNA targeting MTA1 gene into the lentiviral vector.Real time PCR and Western blotting results showed that the expressions of MTA1 mRNA and MTA1 protein in 5-8F cell lines were down-regulated significantly after siRNA transfection.Conclusions MTA1 may promote the malignant transformation and enhance the metastatic potential of nasopharyngeal carcinoma cell lines.The expression of MTA1 in 5-8F cells can be inhibited effectively by MTA1-specific siRNA expression lentiviral vector,which provides a valuable tool for investigating the role of MTA1 gene in the carcinogenesis and progression of nasopharyngeal carcinoma.

Objective:To select the binding-peptide of the cell surface marker CD133 of cancer stem cells from phage peptide library,and to find a new tool for research on stem cells,tumor therapy and anti-metastasis of cancer.Methods:Biotined mouse CD133 extracellular fraction was used as a target to screen phage 7-peptide library by the high affinity of streptavidin and biotin,and the clones were identified by sandwich ELISA and competitive experiment.Single strand DNA was extracted from these positive clones and was analyzed by single-strand dideoxy-sequencing.Results:After three turn solution panning,five peptides with high affinity shared the same amino acid sequence:APSPMIW and three identical peptides with high affinity shared the same amino acid sequence:LQNAPRS.Conclusion:The peptides that bind with mouse CD133 extracellular fraction with high affinity and specificity were first screened from the phage peptide library for the first time,which initially indicates that the feasibility of screening from phage peptide library with small molecule polypeptide biotined as a target.

Background and purpose:The most serious aspect of cancer is the emergence of metastases in organs distant from the primary tumor, since most deaths from cancer are due to metastases. In recent years, a series of studies has been undertaken both in vitro and in vivo, and demonstrated that the CD133+ hematopoietic stem cell is effective in increasing mice tumor cell metastases, but the efficacy of CD133+ stem cells is not well studied for human cancer.We investigated the effects of CD133+ stem cells on the proliferation, adhesion and migration in human colorectal neoplasm cell line SW480.Methods:CD133+ cells were separated from fresh cord blood mononuclear cells(MNC) by Ficoll density gradient centrifugation and magnetic activated cell-sorting system(MACS).The treatment group was the co-culture of CD133+ cells and SW480, the control group used the same culture medium without CD133+ cells. The effect of CD133+ cells on proliferation in SW480 cell was measured by MTT assay, the migration abilities of SW480 cell were assayed in Transwell cell culture, Cell adhesion assay was carried out in a 96 microplate well precoated with fibronection.Results:CD133+ cells could significantly improve the growth ability of SW480 cell, not only the ability of proliferation, but also the morphology. The morphology of the cells was remarkably changed. Irregular nucleus, double nucleus and polymorphic nucleus appeared in the treatment group, and the cells were shaped as polygon-like and leptosomatic. In the cell adhesion assay, A 490 of the treatment group was 0.11?0.01, A 490 of the control group was 0.05?0.01(P﹤0.05).Conclusions:Our results indicate that the CD133+ cell is effective in increasing tumor cell proliferation and invasion. Hematopoietic stem cell may play an important role in the invasion and metastasis of colorectal neoplasm.

OBJECTIVE:To optimize the formulation of Compound pearl powder effervescent granule and study its release behavior in vitro. METHODS:The contents of anhydrous citric acid,sodium bicarbonate,and lactose in the formulation were optimized by uniform design method with the consumption of ethylenediamine tetraacetic acid(EDTA) as index meanwhile giving consideration to the foaming height and pH of the solution. The dissolution of calcium carbonate in Compound pearl powder effervescent granule,self-made calcium carbonate tablets and self-made pearl power in distilled water and hydrochloric acid were investigated. RESULTS:The optimized formulation of the Compound pearl powder effervescent granule was stated as follows:10.0 g anhydrous citric acid,2.5 g sodium bicarbonate,and 5.0 g lactose. There were significant differences across the 3 different preparations in dissolution rates. CONCLUSION:The prepared Compound pearl powder effervescent granule is characterized by fast and complete drug release. Its solution is acid and indicated for specific population.

Purpose　To explore the clinicopathological features of subcutaneous panniculitic T-cell lymphoma(SPTCL) and significances of genetic analysis in the diagnosis. Methods　Histopathology, immunohistochemitry and detection of clonal gene rearrangement by PCR were used in 3 cases of subcutaneous panniculitic T-cell lymphoma (SPTCL), which were originally diagnosed as relapsing nodular nonsuppurative panniculitis. Results　Three misdiagnostic cases were correctly redefined as subcutaneous panniculitic T-cell lymphoma, with immunophenotype of CD45+,CD45RO+, Mac387－,and clonal TCR-β gene rearrangement. Conclusions　Subcutaneous panniculitic T-cell lymphoma has distinctive clinicopathological features. Genetic analysis is an effective method for the diagnosis of SPTCL.

Aim Toinvestigatetheanalgesiceffectsof epidural osthole application on the mechanical allodyn-ia and the ERK/MAPK signaling pathway and the expression of COX-2 mRNA in the spinal dorsal horn.Methods 125adultmaleSDratswererandomizedin-to five groups( n=25 each) :Blank, Sham, NP, Ost and vehicle. At postoperative day 6, 1mg/rat osthole 50 μl was injected epidurally into group Ost and the same volume of vehicle was given into group vehicle. The mechanical pain threshold was measured by 50%MWT at 1 day before operation and the 3 rd,6 th,7 th, 14 th,21 st day after operation. After the measurement of pain threshold on postoperative day 14 , the L4-6 segment of spinal dorsal horn was removed for determi-nation of the expression of ERK, pERK and COX-2 mRNAbyWesternblotandRT-PCR.Results Com-pared with blank group, the mechanical pain threshold was only down-regulated at day 1 after operation in sham group, the expression of pERK and COX-2 mR-NA in sham group showed no significant difference ( P>0. 05 ); the mechanical pain threshold was signifi-cantly down-regulated after operation in NP, Ost and vehicle groups( P<0. 05 ) and the expression of pERK and COX-2 mRNA was significantly increased ( P <0. 05). Compared with vehicle group, the pain thresh-old in Ost group was significantly increased after drug administration( P<0. 05 ) and the expression of pERK and COX-2 mRNA was significantly reduced ( P <0. 05 ) . The expression of ERK showed no significant difference among each group(P>0. 05). The correla-tion analysis on pERK1/2 and COX-2 mRNA revealed the Pearson correlation coefficient was 0 . 878 and 0 . 910 , suggesting a strong positive correlation between pERKandCOX-2mRNA.Conclusions Ostholead-ministrated in the early stage after surgery can alleviate the nucleus pulposus-induced radicular inflammatory pain probably by inhibiting the expression of pERK and COX-2 mRNA in spinal dorsal horn.

Objectlve To demonstrate the feasibility of three-dimensional(3D)spin-lattice relaxation time in the rotating frame(T1ρ)-weighted imaging of porcine patellar cartilage in vitro at 7.0 T and the measurement of T1ρ of agarose phantom and patellar cartilage.Methods All the MR Imaging experiments were performed on a 7.0 T Varian scanner using a 6.0-cm-diameter quadrature birdcage RF coil tuned to 300 MHz.A 3D spin-echo sequence with a self.compensating spin-lock pulse cluster was used to acquire 3D-T1ρ-weighted images.The time of spin-locking(TSL)was from 0 to 50 ms with an interval of 10 ms.Spin-lock power wag 440 Hz.3D-T1ρweighted imaging was done three times for 6 phantoms (concentration 1%t0 6%),as well as once for 8 porcine patellar cartilages in order to assess the reproducibility of this technique.Signal-to-noise ratio(SNR)was measured on the acquired images of both phantoms and patellar cartilages,which were tested for significance using Two-way ANOVA.The images were processed on Vnmr J workstation using home-built processing software to construct 3 D T1ρ maps.T1ρ values were calculated within manually drawn regions-of-interest(ROI),and differences between groups were tested for significance using analysis of variance(One-way ANOVA).Results T1ρ -weighted images with a shorter TSL had a higher SNR value,which measured between 48±8 and 95±8 in the global cartilage.Cartilage images had a higher SNR(TSL＜30 ms)compared to agarose phantoms and a lower SNR(TSL ＞30 ms)only compared to l%agarose phantorm T1ρ relaxation times in agarose phantoms increased as agarose concentrations decreased in global regions.The CV of T1ρ in agarose phantoms was less than 10%.Global and regional analyses of patellar cartilage T1ρ were 68.9±6.3 ms,80.7±12.8 ms,65.7±7.0 ms,82.4±7.7 ms,and 69.7±6.4 ms in the global cartilage,the superficial layer,the transitional layer,the deep layer,and the calcified layer,respectively.T1ρ in the superficial and deep layer was significantly higher than in the transitional,calcified layer and global cartilage(F=6.436,P＜0.05).Conclusions The present study demonstrates the feasibihty of 3D-T1ρ-weighted imaging of porcine patellar cartilage at 7.0 T with hish image quality.T1ρ maps can be used to quantify the laminar structures in 3D-T1ρ-weighted images of articular cartilage,which pave the way to evaluate early osteoarthritis and cartilage regeneration.

OBJECTIVETo investigate the expression of inhibitor of DNA differentiation/DNA binding 1 (Id1) and Id3 in endometrial carcinoma and explore their roles in regulating the proliferation, invasion, migration and adhesion of endometrial carcinoma cells in vitro.

METHODSId1 and Id3 expression in 4 fresh endometrial cancer tissue specimens and matched adjacent tissues were detected using Western blotting. Two endometrial cancer cell lines, HEC-1-B and RL-952, were both divided into 4 groups, namely the untreated group, blank virus group, promoter group and Id1/Id3 double-knockdown group, and their expressions of MMP2, CXCR4 and P21 were detected by qRT-PCR and Western blotting. The proliferation, invasion, migration and adhesion of the cells were evaluated with MTT, Transwell, wound-healing, and adhesion assays.

RESULTSEndometrial carcinoma tissues showed significantly higher Id1 and Id3 expression than the adjacent tissues (P<0.05). In the two endometrial carcinoma cell lines, Id1/Id3 double-knockdown significantly decreased MMP2 and CXCR4 expression and increased P21 expression at both mRNA and protein levels (P<0.05), and resulted in suppressed cell proliferation, invasion, migration and adhesion.

CONCLUSIONId1 and Id3 expressions are up-regulated in endometrial carcinoma to promote the proliferation, invasion, migration and adhesion of the tumor cells by increasing MMP2 and CXCR4 expression and reducing P21 expression. Therapies targeting Id1/Id3 can be a novel strategy for treatment of endometrial carcinoma.

Cell Adhesion ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Endometrial Neoplasms ; metabolism ; pathology ; Female ; Humans ; Inhibitor of Differentiation Protein 1 ; metabolism ; Inhibitor of Differentiation Proteins ; metabolism ; Matrix Metalloproteinase 2 ; metabolism ; Neoplasm Proteins ; metabolism ; Proto-Oncogene Proteins p21(ras) ; metabolism ; RNA Interference ; Receptors, CXCR4 ; metabolism

OBJECTIVETo investigate the correlation between the expressions of glypican-3 (GPC3) and Notch1 and their roles in the tumorigenesis and progression of hepatocellular carcinoma (HCC).

METHODSImmunohistochemistry and computerized image analysis were utilized to quantitatively detect the expressions of GPC3 and Notch1 in 30 HCC tissue specimens.

RESULTSIn the 30 HCC specimens, GPC3 expression decreased significantly as the grade of tumor differentiation increased (P<0.05 or P<0.01), while Notch1 expression presented with a reverse pattern of changes (P<0.05 or P<0.01). An obvious negative correlation was found between the expressions of GPC3 and Notch1 in HCC tissues (rp=-0.607, P=0.000; r=-0.692, P=0.000).

CONCLUSIONThe expressions of GPC3 and Notch1 show a negative correlation in HCC, suggesting a possible mechanism for mutual regulation between them to contribute to the tumorigenesis and progression of HCC.

Adult ; Aged ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Female ; Glypicans ; metabolism ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Male ; Middle Aged ; Receptor, Notch1 ; metabolism

Objective:To evaluate the efficacy and safety of palonosetron in preventing chemotherapy-induced vomiting. Meth-ods:A multi-center, randomized, double-blind, and self-cross-over positively controlled clinical trial design was used. All patients were randomized into two groups, as follows:Regiment A (61 cases) and Regiment B (64 cases). Regimen A with palonosetron hydrochlo-ride injection (test agent) was used in the treatment cycle A, whereas granisetron hydrochloride injection (control drug) was used in the cycle B. Treatments were randomly administered on the patients of the two groups. Regimen B was on the contrary, the control drug was used in the cycle A, and the test agent was used in the treatment cycle B. All patients treated with the test agent were classified as the test group, whereas those treated with the control drug were classified as the control group. Complete control rate and adverse reac-tion of acute and delayed vomiting in the two groups during the two cycles of chemotherapy regimen were compared. Results: In Group One, the complete control rate of delayed vomiting was significantly higher in the palonosetron administration cycles than in the granisetron cycles (76.92%vs. 55.38%, P=0.0110). In the same group, the frequency of vomiting was significantly less in palonosetron cycles than in the granisetron cycles during day 1 to day 5 (1.32±3.42 vs. 1.94±3.03, P=0.0096). The incidences of adverse effects were low in both groups. No grades 3 and 4 adverse effects were observed. Conclusion: Palonosetron showed efficacy in preventing the acute and delayed chemotherapy-induced vomiting. The drug is superior to granisetron, specifically in delaying vomiting in Group One. Palonosetron hydrochloride showed slight adverse effects. Hence, this drug can be used in clinic.