Promotion of proliferation and metastasis of hepatocellular carcinoma by LncRNA00673 based on the targeted-regulation of notch signaling pathway.

OBJECTIVE: To investigate the relative expression of long non-coding RNA 00673 (lncRNA 00673) in hepatocellular carcinoma (HCC) and HCC cells and study its regulation on the malignant phenotype of HCC cells PATIENTS AND METHODS: Samples of HCC and adjacent tissues from January 2013 to December 2015 were collected. The expression level of lncRNA00673 in HCC tissues and cells was detected by quantitative Real-time polymerase chain reaction (qRT-PCR) assays. lncRNA00673 specific interference sequences were transiently transfected into HCC cells and the effect of HCC cells on the biological behavior of HCC cells was examined by in vitro experiments ((3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT assay), flow cytometry, transwell, etc.). A tumor model of nude mice with HCC was established for the study of tumor growth condition of tumor-bearing mice after the interference with lncRNA00673 expression. Changes in expression levels of molecular markers on Notch signaling pathway after the interference with lncRNA00673 were detected by Western blot. RESULTS: lncRNA00673 was highly expressed in HCC tissues and cells. MTT results showed that interfering with lncRNA00673 inhibited cell proliferation. Flow cytometry results showed that HCC cell cycle was retarded in G1-G0 phase, thus promoting apoptosis after the interference with lncRNA00673. Western blot results showed that expression levels of molecular markers on Notch signaling pathway were changed after the interference with lncRNA00763. CONCLUSIONS: lncRNA00673 is highly expressed in HCC tissues and cells, and can promote the proliferation and metastasis of HCC by the regulation on Notch signaling pathway. lncRNA00673 may be a potential target for the treatment of HCC.

Occurrence and Genetic Diversity of Grapevine berry inner necrosis virus from Grapevines in China.

To investigate the prevalence and genetic diversity of Grapevine berry inner necrosis virus (GINV) in China, 195 grapevine samples from 15 Chinese provinces and regions were tested using reverse-transcription polymerase chain reaction. The samples included symptomatic and asymptomatic cultivars, with 35.9% (70 of 195) of samples testing positive for GINV. Seventeen samples had obvious ring spot symptoms, and 94.1% (16 of 17) tested positive for GINV, suggesting that GINV may be highly associated with the ring spot symptom. The genetic diversity of GINV isolates was analyzed based on the partial nucleotide and amino acid sequences of the coat protein (CP) and movement protein (MP) genes. Phylogenetic analyses of the MP and CP gene sequences divided the GINV isolates into three groups. The majority of the Chinese isolates were in groups 1 and 2, and only one Chinese isolate, along with a previously reported Japanese isolate, was in group 3. This is the first report on the genetic diversity of GINV isolates and their prevalence and distribution in China.

Detection and sequence analysis of grapevine virus B isolates from China.

The presence of grapevine virus B (GVB) was detected in 188 grapevine samples from China by double antibody sandwich ELISA (DAS-ELISA) and reverse transcription-PCR (RT-PCR). The accuracy of detection by RT-PCR was confirmed by sequencing amplified PCR fragments. Seventeen samples were GVB-positive by DAS-ELISA and five by RT-PCR. The isolate QMW proved to be positive by RT-PCR only, and four isolates (DGWH, DGW, QM, and JFL) could be detected by both methods. Among the five GVB-positive samples detected by RT-PCR, two isolates were originally collected from Henan province and three from Liaoning province. The expected 722 bp DNA fragment, covering partial ORF3 through partial ORF5, was amplified from the five GVB infected samples. Sequence analysis revealed that the molecular variants΄ composition of GVB in the different isolates was complex. Clones of DGWH, DGW, QM, and JFL isolate shared high nucleotide identities, while the identities among the clones of isolate QMW varied. The variants of GVB isolates obtained in this study showed nucleotide identities from 81.1% to 97.9% among themselves, and 79.1% to 98.5% identity with five previously published GVB isolates in NCBI. The alignment of partial ORF3 and the phylogenetic relationships of ORF4 revealed that the molecular variants of Chinese GVB isolates could be clustered into three groups. Only isolate DGW was in the same group with the reported GVB isolates from other countries; the other four GVB isolates in this study were clustered into two groups.

Persistent toxic substances in agricultural soils of Lishui County, Jiangsu Province, China.

The contamination status and distribution of PTS, including HCHs, DDTs, PCBs, and PAHs, were studied in the agricultural soil of Lishui County, one of China's nationally designated eco-demonstration region. The PTS was analyzed by GC, GC/MS, and HPLC coupled with microwave extraction. The concentrations of the sum of HCHs, DDTs, and PCBs ranged from <0.1 ng g(-1) to 133 ng g(-1), 93.8 ng g(-1), and 81.0 ng g(-1) (dry wieght), respectively; while the contents of the sum of PAHs ranged from 3.26 ng g(-1) to 91.0 ng g(-1) (dry wieght). The soil investigation revealed that there were no recent HCHs and DDTs pollution; none of the soil PCBs concentrations suggested a serious biological threat in the investigated area; the extent of pollution of PAHs was low compared to other investigations. In summary, levels of PTS contamination in the soil of the Lishui County were relatively low, but caution should be taken in industrial zone.

PbZr0.4Ti0.6O3 and Ba0.9Sr0.1TiO3 reflectors derived from chemical solutions containing polymers.

Ba(0.9)Sr(0.1)TiO3 (BST)-based and PbZr(0.4)Ti(0.6)O(3)-based quasi-periodic multilayers consisting of dense and porous ferroelectric layers have been fabricated by solgel technique using chemical solutions containing polyethylene glycol (PEG) or polyvinylpyrrolidone k30 (PVP). All multilayers exhibit good performance as dielectric mirrors. For each multilayer, the maximum peak reflectivity is over 90% and the photonic stopband width is no less than 30 nm at room temperature. The reflection-band position can be easily tuned by varying the thickness of the bilayer. With the same processing conditions and number of periods, the Bragg reflection performance is almost the same for quasi-periodic PZT multilayers derived from two precursors containing different polymers. The BST multilayers deposited by using a PVP-containing precursor are superior in optical properties, including peak reflectivities and stop-band width, to those deposited by using the PEG-containing solution.

Characterization of the humoral and cellular immune responses against hepatitis C virus core induced by DNA-based immunization.

Hepatitis C Virus (HCV) causes most cases of posttransfusion hepatitis. Chronic HCV infection is highly related to chronic hepatitis, cirrhosis and hepatocellular carcinoma. Current therapies are only minimally effective and no vaccine has been developed. DNA-based immunization could be of prophylactic and therapeutic value for HCV infection. By intramuscular inoculation in BALB/c mice with an HCV recombinant plasmid pCI-HCV-C, we found significant levels of IgM antibody, but no significant IgG rise. After boost the immunized mice with recombinant HCV-core protein (cp1-10; 1-164aa), the anticore IgG, verified by Western-blotting, rose rapidly, which was two weeks earlier than that with control plasmid. Spleen cells from pCI-HCV-C immunized mice gave higher proliferation index (PI) than control (P < 0.05). The PI of cp1-10 boosted mice was even higher. Proliferation blocking assay with mAb proved the responding cell to be of CD4+ CD8- phenotype, supporting specific priming of T helper cells. A 51Cr-releasing CTL assay specific for HCV-core was developed, and a specific CTL response against HCV-core was demonstrated in both pCI-HCV-C immunized mice and mice boosted with cp1-10. Strong cytotoxic activity against peptide-pulsed p815 cells (H-2d), but not EL-4 cells (H-2b), suggested MHC class I restriction of the CTL activity. Blocking of CTL with mAb proved the effector cells to be of CD4- CD8+. Three CTL epitopes in HCV-core protein were demonstrated. We failed to detect CTL when immunized only with core protein. The results suggested that vaccination with HCV-core derived DNA sequences could be an effective method to induce humoral and cellular immune responses to HCV.

[Calcium distribution changes during epididymal maturation of mouse and guinea pig sperms].

Calcium was localized by in situ precipitation with potassium antimonate during epididymal maturation of the mouse and guinea pig sperms. In caput epididymis the calcium in mouse sperm head was mainly localized on the inner surface of tha outer acrosomal membrane (OAM) in preacrosomal region. During the passage from the caput epididymis to the cauda epididymis the calcium amount of mouse sperm did not undergo apparent changes. In comparison with mouse sperm, there were a few fine calcium deposite granules on the inner surface of the guinea pig sperm OAM on the abdomen side at the caput epididymis stage, but these granules disappeared at the cauda epididymis stage. The microvilli of columnar cell in columnar epithlium lining the epididymal duct are probably involved in regultion of Ca2+ concentration of the intraluminal fluid. The calcium precipitation granules distributing in the microvilli were observed. An abundance of Ca2+ were present in the intraluminal fluid of the corpus epididymis and they might have some important functions during the process of sperm maturation. Calcium in sperm tail was mainly distributed in the mitochondria. Compared with the mouse sperm, the mitochondria of guinea pig sperm possessed more calcium.

Studies on a new antiarrhythmic drug changrolin-4-(3',5'-bis [(N-pyrrolidinyl) methyl]-4'-hydroxyanilino)-quinazoline.

Changrolin is 4-(3', 5'-bis[(N-pyrrolidinyl)methyl]-4'-hydroxyanilino)-quinazoline. It is a novel type of antidysrhythmic drug. It has been synthesized in 4 steps. According to our experiments, changrolin exhibited significant protective and therapeutic effects against experimental arrhythmias induced by aconitine or ouabain. It raised the electrical threshold of ventricular fibrillation. Intravenous injections in dogs and rabbits caused (i) a mild tachycardia followed by bradycardia; (ii) a prolongation of P-R interval and a widening of QRS complex in the electrocardiogram; (iii) a gradual hypotehsion; (iv) a slight weakening of cardiac functions; and (v) only moderate influences on the hearts of dogs and rabbits when the rate of infusion was less than 1 mg/min. Changrolin could be well absorbed by oral administration. Absorption appeared to be more rapid and complete by intramuscular injection. 14C-labelled changrolin was distributed mainly in the liver and the alimentary tract.