Objective To explore the expression of GPR30,HRG1 and HER2 including the activation status of HER2 (phosphorylated HER2)in invasive ductal breast cancers and their relationship with lymphatic metastasis.Methods The expres-sion of GPR30,HRG1,HER2 and pHER2 in 72 cases of specimens of invasive ductal breast cancers were examined by immunohis-tochemistry method.Results A moderate correlation between GPR30 and HRG1 was disclosed (r=0.597,P =0.000).There was strong correlation between pHER2 and GPR30 or HRG1(r=0.742,P =0.000;r=0.615,P =0.000).The expression of GPR30 and pHER2 in the lymphatic metastasis group was remarkably higher than in the group without lymphatic metastasis(P <0.05). Conclusion The interaction between GPR30 and HRG1 HER2 signal transduction pathways might be involved in the lymphatic metastasis in breast cancer.Blocking both of GPR30 and HRG1 signaling pathway could be a promising new strategy for breast cancer treatments.

Objective To evaluate the value of 18F-FDG and 18F-FLT PET/CT for the detection of primary and regional lymph node metastasis of gastric cancer.Methods Thirty-seven patients with gastric cancer underwent preoperative 18F-FLT and 18F-FDG PET/CT within one week from March 2011 to April 2013.Postoperative histopathology confirmation was obtained in all patients.The PET/CT images were assessed visually and semi-quantitatively.Two-sample t and x2 tests were analyzed using SPSS 13.0.Results For the detection of primary gastric cancer,the sensitivities of 18 F-FLT and 18 F-FDG PET were 89.2％ (33/ 37) vs 91.9％(34/37),respectively (x2=0.158,P＞0.05).The 18F-FLT SUVmax of 16/37 cases with diffuse-type gastric cancer was significantly higher than that of 21/37 cases with intestinal-type gastric cancer (6.89±1.38 vs 3.79±2.45,t=4.533,P＜0.05) ; while 18F-FDG SUVmaxwas not significantly different between the two subgroups (7.13± 1.97 vs 6.36±2.32,t =1.066,P＞0.05).For the detection of regional lymph node metastasis,the sensitivity,specificity and accuracy of 18F-FLT and 18F-FDG were 64.8％(35/54) vs 88.9％(48/54),97.6％(246/252) vs 82.9％(209/252),91.8％(281/306) vs 84.0％(257/306),respectively (x2 =8.796,30.948,8.854,all P＜0.05).The overall sensitivity,specificity and accuracy by both tracers were 92.6％(50/54),98.8％(249/252) and 97.7％(299/306).Conclusions 18F-FLT might be a better or complementary tracer to 18F-FDG for the detection of diffuse-type gastric cancer.Compared with 18FFDG PET/CT,18F-FLT PET/CT may be less sensitive but more specific and accurate for the detection of regional lymph node metastasis.The overall diagnostic accuracy can be improved by using both tracers.

Objective To investigate the undergraduates′cognition ,attitude and behavior about low‐carbon life and then to provide a relatively scientifically strategy .and provide scientific basis to formulate countermeasures about undergraduates′low‐car‐bon lifestyle .Methods By multistage sampling ,382 students′knowledge ,attitude and practice about low‐carbon life in Wuhan Uni‐versity of science and technology were analyzed .Results The students′cognition about low‐carbon life was good ,the awareness rate of girl was higher than that of boy ;Their attitude was positive ,grade and knowledge awareness were the two influence factors ;the behavior situation was unsatisfactory ,the origin of students and attitude enthusiasm were the elements affect rationality of be‐havior .Conclusion Undergraduates should strengthen the cognition of low‐carbon life ,improve attitude enthusiasm ,and form a good behavioral habit of low‐carbon life .

This study aimed to explore Semaphrin4D (Sema4D) expression and clinical significance in non-small cell lung cancer (NSCLC), and to define the roles and mechanisms of Sema4D in regulating the malignant behaviors of A549 cells by small interfering RNA (siRNA). Firstly, immunohistochemistry revealed that Sema4D was more frequently expressed in NSCLC than in lung benign lesion (P<0.05) and its overexprssion was associated with low differentiation (P<0.05), poor pTNM staging (P<0.05) and occurrence of lymph node (LN) metastasis (P<0.05). Endogenous Sema4D expression was suppressed by Sema4D siRNA in A549 cells overexpressing Sema4D. Protein levels of Sema4D, total Akt and p-Akt were examined by Western blotting. Cell proliferation, migration and invasion abilities were measured by MTT assay and Transwell assay respectively. Results showed that Sema4D siRNA significantly suppressed phosphorylation of AKT in A549 cells, but it did not alter total AKT expression. In addition, efficient down-regulation of SemaD significantly inhibit cell proliferation (P<0.05), migration (P<0.05) and invasion (P<0.05) in A549 cells. These findings suggest that Sema4D might serve as a reliable tool for early prediction of NSCLC poor prognosis. Sema4D could play an important role in promoting tumor proliferation, migration and metastasis in the NSCLC, by influencing the Akt protein phosphorylation. Inhibition of Sema4D may be a useful approach for the treatment of NSCLC.

Objective To investigate the value of MRI diffusion-weighted imaging (DWI) technique in endocrine therapy for prostate cancer (PCa) based on PI-RADSv2.1. Methods A retrospective analysis of 57 patients with pathologically confirmed PCa was conducted. All patients underwent multi-parametric MRI (mpMRI) according to PI-RADS v2.1 technical specifications before biopsy and six months after endocrine therapy. The apparent diffusion coefficient (ADC) values were measured in cancer and non-cancer areas before biopsy and six months after endocrine therapy. Patients were grouped based on the mRECIST criteria and PSA level into responders (n=45) and non-responders (n=12). ROC curves were obtained to assess the correlation between changes in ADC values and PSA values before and after endocrine therapy. Results In the responder group, the ADC value of the cancer areas was increased significantly after endocrine therapy (P<0.001). No statistically significant difference of the ADC value of the cancer areas was found in the non-responder group before and six months after endocrine therapy (P=0.714). The ADC change of responders and non-responder groups were (0.411±0.178)×10^-3 mm²/s and (-0.014±0.125)×10^-3 mm²/s, respectively (P<0.001); the ADC ratio were (60.603±30.201)% and (-1.096±13.175)%, respectively (P<0.001). The cutoff value of the ADC change was 0.165 (AUC=0.974; sensitivity, 88.89%; specificity, 100.00%; PPV, 100.00%; NPV, 70.59%). The cutoff value of ADC ratio was 16.827% (AUC=0.980; sensitivity, 91.11%; specificity, 100.00%; PPV, 100.00%; NPV, 75.00%). The ADC values were negatively correlated with serum PSA before and after endocrine therapy. Conclusion The ADC change and ADC ratio may be facilitated to monitor the efficacy of endocrine therapy for PCa. The ADC values were negatively correlated with serum PSA.

OBJECTIVETo study the constituents from roots of Euphorbia hylonoma.

METHODColumn chromatographic techniques were used for isolation and purification of the chemical constituents and their structures were identified by spectral analysis (IR, 1H-NMR, 13C-NMR, 2D-NMR and MS).

RESULTSix compounds were isolated and elucidated as nonane (1), bis (2-ethylhexyl) phthalate (2), euphol (3), beta-sitosterol (4), acalyphol (5) and daucosterol (6) respectively.

CONCLUSIONCompounds 1, 2, 3, 5 and 6 were isolated from the plant for the first time.

Alkanes ; chemistry ; isolation & purification ; Diethylhexyl Phthalate ; chemistry ; isolation & purification ; Euphorbia ; chemistry ; Lanosterol ; analogs & derivatives ; chemistry ; isolation & purification ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Sitosterols ; chemistry ; isolation & purification ; Triterpenes ; chemistry ; isolation & purification

BACKGROUNDTo select the mutants of HPV type 16 E6 and E7 genes suitable for construction of vaccine for treatment of cervical cancer.

METHODSE6 and E7 genes were modified by site-directed mutagenesis. Several recombinant vaccina viruses were constructed by inserting the E6 or E7 mutants into the genome of vaccina virus Tiantan strain and employed to study their antigenicity.

RESULTSWestern blot assay showed that the E6 ?mutant? with substitution of Gly for Leu at amino acid site 50 and E7 mutant with substitution of Gly for Cys-24 and Glu-26 had no effect on their stability and antigenicity, but change of the Cys at position 91 of E7 dramatically reduced its stability and antigencity. Conclusion The results confirmed that the Zinc-finger structure at the E7 C-terminal? plays an important role in the integrity and stability of E7 protein.

Animals ; Antibodies, Viral ; biosynthesis ; Female ; Mice ; Mice, Inbred C57BL ; Mutagenesis, Insertional ; Oncogene Proteins, Viral ; genetics ; immunology ; Papillomaviridae ; genetics ; Papillomavirus E7 Proteins ; Repressor Proteins ; Vaccinia virus ; immunology ; Zinc Fingers

OBJECTIVETo investigate the expression profiles of myocardin gene during the differentiation of bone marrow-derived mesenchymal stem cell to smooth muscle cells in the conditional medium combined with a high concentration of fetal bovine serum (FBS).

METHODSMarrow-derived mesenchymal stem cells were isolated and purified from mouse femoral bone and shinbones using differential adherent methods. Cells at the third passage were induced by 20% FBS in conditioned medium, conditioned medium alone, 20% FBS or 10% FBS alone respectively. Mouse aortic smooth muscle cells were cultured as the positive control. Levels of mRNA and protein expression of myocardin and several smooth muscle cells marker genes were determined by immunofluorescence, RT-PCR and Western blot before and 3, 7, 10, 14 d after the induction. The presence of smooth muscle myofilaments was detected by using transmission electron microscope.

RESULTSNaive bone marrow-derived mesenchymal stem cells displayed multiple morphological forms including fusiform, polygon, oval, and micro-spherical, as compared to the single macro-spindle form after the induction. Typical appearance of peak valley was displayed on the 21st day after induction. At the same time, the expression of smooth muscle marker genes was reinforced along with an up-regulation of myocardin expression. Immunofluorescence study showed that the cells expressing myocardin and smooth muscle marker genes such as alpha-SMA and SM-MHC increased. Fluorescence domain of myocardin translocated from cytoplasm to nucleus and the amounts of double positive cells for myocardin with alpha-SMA or SM-MHC also increased. RT-PCR confirmed that the mRNA expression of myocardin increased gradually and remained stabilized after achieving its peak on the 7th day after induction. The expression of smooth muscle marker genes, alpha-SMA and SM22alpha, remained stable on the 10th day of induction. It was also confirmed by Western blot that the protein expression of both myocardin and alpha-SMA were markedly increased during the induction. Finally, transmission electron microscopy revealed the presence of myofilament on the 21st day after induction.

CONCLUSIONSBone marrow-derived mesenchymal stem cells can be effectively induced into smooth muscle-like cells by conditioned medium combined with 20% FBS. Myocardin plays an important role in the differentiation process of bone marrow-derived mesenchymal stem cells to the peripheral smooth muscle cells.

Animals ; Bone Marrow Cells ; cytology ; physiology ; Cattle ; Cell Differentiation ; physiology ; Male ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Mice ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; physiology ; Nuclear Proteins ; genetics ; metabolism ; physiology ; Reverse Transcriptase Polymerase Chain Reaction ; Trans-Activators ; genetics ; metabolism ; physiology ; Up-Regulation

OBJECTIVETo generate a candidate HPV16 vaccine for experimental and therapeutical use for cervical cancer.

METHODSThe mutants of HPV16 early E6 and E7 genes were inserted into a vaccinia virus expression vector. A strain of recombinant vaccinia virus was constructed through homologous recombination.

RESULTSShowed that the mutant E6 and E7 genes were located at TK gene region of vaccinia virus Tiantan strain in a head to head orientation under the control of early/late promoters, H6 and 7.5K respectively. Studies in mice indicated that VmE6E7 could elicit specific antibodies against E6 and E7, and retarded or prevented tumor development in a proportion of C57 BL/6 mice challenged by syngeneic HPV16E6 and E7 transformed tumor cells.

CONCLUSIONSThe success in constructing VmE6E7 provides a basis for the further development of HPV16 therapeutic vaccine.

Animals ; Female ; Genes, Viral ; genetics ; Genetic Vectors ; genetics ; immunology ; Mice ; Mice, Inbred C57BL ; Mutation ; Neoplasms, Experimental ; prevention & control ; Oncogene Proteins, Viral ; genetics ; Papillomaviridae ; genetics ; Papillomavirus E7 Proteins ; Recombination, Genetic ; Repressor Proteins ; Transfection ; Vaccinia virus ; genetics ; immunology

OBJECTIVETo generate an HPV16 prophylactic vaccine candidate for cervical cancer.

METHODSHPV16 major capsid protein L1 gene and minor capsid protein L2 gene were amplified using PCR. These genes were mutated by PCR site-directed mutagenesis for removal of sequence motifs (TTTTTNT) which would cause transcription termination when expressed from a vaccinia virus early promoter, then inserted into a vaccinia virus expression vector. A strain replication-deficient recombinant vaccinia virus containing the mutant sequences was obtained through a homologous recombination and identified.

RESULTSThe nucleotide sequence remained the correct amino acid sequence of the L1 and L2 proteins after mutated. Full-length L1 and L2 proteins were generated in cells infected with the recombinant virus. The virus strain propagated at very low titer or could not reproduce in some kinds of cell derived from different human tissues.

CONCLUSIONSThe authors have generated a strain replication-deficient recombinant vaccinia virus expressing HPV16 L1 plus L2 proteins as an HPV16 prophylactic vaccine candidate for cervical cancer.

Capsid ; Capsid Proteins ; genetics ; Cell Line ; Cloning, Molecular ; Female ; Gene Expression ; Genetic Vectors ; Humans ; Oncogene Proteins, Viral ; genetics ; Papillomaviridae ; genetics ; physiology ; Papillomavirus Infections ; prevention & control ; Transfection ; Tumor Virus Infections ; prevention & control ; Uterine Cervical Neoplasms ; virology ; Vaccinia virus ; genetics ; physiology ; Virus Replication