CalliSpheres® drug-eluting beads (DEB) transarterial chemoembolization (TACE) is equally efficient and safe in liver cancer patients with different times of previous conventional TACE treatments: a result from CTILC study.

PURPOSE: To assess the efficacy and safety of drug-eluting beads transarterial chemoembolization (DEB-TACE) in liver cancer patients with different times of previous conventional transarterial chemoembolization (cTACE) treatments. METHODS: 367 liver cancer patients about to receive DEB-TACE treatment were enrolled in this prospective cohort study. All patients were divided into no previous cTACE group (NPC group), 1-2 times previous cTACE group (PC group) and triple or above previous cTACE group (TPC group) according to the times of previous cTACE treatments. RESULTS: There was no difference in complete response (CR) (P = 0.671) and objective response rate (ORR) (P = 0.062) among three groups. Additionally, no difference in overall survival (OS) among groups (P = 0.899) was found. As to liver function, most liver function indexes were deteriorative at 1 week after DEB-TACE operation, but returned to baseline at 1-3 months after DEB-TACE operation in all three groups, while percentage of abnormal total bile acid (TBA) patients was higher in TPC group than NPC and PC groups at 1-3 month post-DEB-TACE (P = 0.018). As for safety profiles, the incidence of pain during DEB-TACE operation was lower in TPC group compared to NPC and PC groups (P = 0.005), while no difference of other adverse events was found during and 1 month post-DEB-TACE treatment among three groups. CONCLUSION: DEB-TACE treatment was equally efficient and tolerated in liver cancer patients with different times of previous cTACE treatments.

MYBA2 gene involved in anthocyanin and flavonol biosynthesis pathways in grapevine.

MYBA2 transcription factor (Myb-related gene) affects the coloring in grapevine berry and plays an important role in the biosynthesis of anthocyanin. The MYBA2 gene was cloned from Vitis vinifera L. cv. Cabernet Sauvignon and polyclonal antibodies for VvmybA2 were prepared. The VvmybA2 gene expression patterns were observed in seven tissues (the leaf, stem, flower, bud, root, berry, and tendril) and during the berry development stage at transcriptional and translational levels, respectively. The results indicated that the expression of VvmybA2 was approximately 11-fold higher in the berry than that in the other six tissues, and increased rapidly from 60 days after full bloom reaching a maximum on day 80. Furthermore, both the anthocyanin content and UDP-glucose:flavonoid-3-O-glucosyltransferase (UFGT) gene expression levels increased rapidly 60 days after full bloom. Moreover, correlation analysis indicated that the transcriptional and translational expression levels of the VvmybA2 gene were significantly positively correlated with not only UFGT and DFR genes but also with the anthocyanin content during berry development. These results suggested that VvmybA2 could not only regulate the transcription of both UFGT and DFR but also is involved in the expression of the UFGT gene associated with color determination in grape berries.

Genetic variation and genetic structure of the endangered species Sinowilsonia henryi Hemsi. (Hamamelidaceae) revealed by amplified fragment length polymorphism (AFLP) markers.

Comprehensive research of genetic variation is crucial in designing conservation strategies for endangered and threatened species. Sinowilsonia henryi Hemsi. is a tertiary relic with a limited geographical distribution in the central and western areas of China. It is endangered because of climate change and habitat fragmentation over the last thousands of years. In this study, amplified fragment length polymorphism markers were utilized to estimate genetic diversity and genetic structure in and among S. henryi. In this study, Nei's genetic diversity and Shannon's information index were found to be 0.192 and 0.325 respectively, indicating a moderate-to-high genetic diversity in species. According to analysis of molecular variation results, 32% of the genetic variation was shown to be partitioned among populations, demonstrating a relatively high genetic divergence; this was supported by principal coordinate analysis and unweighted pair-group method with arithmetic average analysis. Moreover, the Mantel test showed that there was no significant correlation between genetic and geographical distances. The above results can be explained by the effects of habitat fragmentation, history traits, and gene drift. Based on the results, several implications were indicated and suggestions proposed for preservation strategies for this species.

Accumulation of flavanols and expression of leucoanthocyanidin reductase induced by postharvest UV-C irradiation in grape berry.

To examine the effect of postharvest ultraviolet C (UV-C) irradiation on flavanol polyphenol accumulation in the grape berry, we investigated total flavanol polyphenol content, the enzyme activity of leucoanthocyanidin reductase (LAR), and transcription of Vv lar1 and Vv lar2 using spectrophotometry, real-time polymerase chain reaction, and western blot analysis in 5-year-old Vitis vinifera L. cv. Cabernet Sauvignon plants. Our results indicated that the accumulation of flavanol polyphenol reached its highest value when exposed to UV-C irradiation for 30 min. Additionally, UV-C irradiation induced the transcription of Vv lar1 and Vv lar2 and the synthesis of LAR1 and LAR2 proteins, resulting in increased accumulation of flavanol polyphenol in the grape berry. Moreover, these effects were associated with the length of time of UV-C irradiation.

In vitro embryo rescue culture of F1 progenies from crosses between different ploidy grapes.

Crossing different ploidy grapes is an effective way to obtain new seedless cultivars. Although embryo rescue has been extensively applied in breeding seedless and triploid grapes, only a few improved cultivars have been developed. Based on preliminary studies, we set five crosses between tetraploid and diploid grape varieties to obtain new hybrid triploid germplasms. Additionally, we compared two different methods of performing in vitro embryo rescue and sowing in the development of hybrid triploid grape plants. The results showed that the germination rate of hybrid seeds was much lower (0-22.8%) than that of the self-pollinated seeds (50.9-61.2%) obtained though the same method of in vitro culture. Meanwhile, the seed germination rates of all crosses obtain through in vitro culture (0-61.2%) were higher than those obtained through sowing (0-42.0%). Identification of ploidy level confirmed that three lines obtained from the crosses of 'Ruby Seedless (2x) x Black Olympia (4x)' and 'Big black (2x) x Kyoho (4x)' were triploid, and one line from the cross of 'Big black (2x) x Kyoho (4x)' was haploid, and the others were diploid, tetraploid, or aneuploidy plants. Moreover, 4 haploid and 42 triploid surviving grape seedlings were planted in a vineyard after propagation. Therefore, an efficient system of breeding triploid seedless grapes using embryo rescue was established and 42 hybrid triploid germplasms were obtained for use in future studies.

Differential expression of resistance to powdery mildew at the early stage of development in wheat line N0308.

Powdery mildew, caused by Blumeria graminis f. sp tritici (Bgt) is one of the devastating diseases of wheat and causes yield losses in temperate wheat growing regions. A wheat line, N0308 with resistance to powdery mildew was used in this study. A suppression subtractive hybridization cDNA library was constructed from the wheat leaves inoculated by Bgt at the two-leaf stage. The differentially expressed genes in response to Bgt infection in wheat were identified, and a total of 175 positive clones from the library were sequenced, and 90 expressed sequence tags (ESTs) were subjected to clustering, BLAST alignment, functional annotation, and classification into different categories. By comparing the EST sequences among the SSH-cDNA libraries, we analyzed gene expression patterns of 7 ESTs associated with the resistance reaction of powdery mildew by using semi-quantitative reverse transcription-polymerase chain reaction. The expression of 5 genes (sulfatase, pathogenesis-related protein 17, betacarbonic anhydrase 2, thioredoxin h-like protein, and coronatine-insensitive) transcripts was induced, and the transcript levels of these genes were the highest at 72 h after Bgt infection, while those of 2 genes (violaxanthin de-epoxidase and gag-pol-polyprotein) were the highest level at 12 and 18 h post-infection, respectively. These findings suggest that these genes are induced at an early stage of infection and are transcriptionally activated for the host defense response.

Higher frequency of CD4(+)CD25(high) Treg cells in hemophilia patients with factor VIII inhibitor.

The production of factor VIII inhibitor antibodies remains the most costly and serious complication in replacement therapy of hemophilia A. We investigated the clinical significance of CD4(+)CD25(high) T regulatory (Treg) cells in hemophilia patients. Our trial included 6 severe hemophilia A patients with factor VIII inhibitors, 6 hemophilia patients without inhibition of factor VIII, and 6 healthy persons (controls). Plasma factor VIII: c was measured by clotting assay. Peripheral blood samples were examined using mutiparameter flow cytometry with fluorescent-labeled monoclonal antibodies. Plasma levels of IFN-γ, IL-2, IL-10, and TGF-ß were measured by ELISA. The frequency of CD4(+)CD25(high) Treg cells in CD4(+) cells was 1.07 ± 0.38% in inhibitor patients and 0.57 ± 0.14% in non-inhibitor patients. The proportion of Treg cells in healthy controls was similar to that of the non-inhibitor patients. However, there were significant differences between the inhibitor and non-inhibitor patients in levels of IFN-γ, IL-2, IL-10, and TGF-ß. We conclude that the proportions of Treg cells and the concentrations of T cell cytokines in inhibitor patients are higher than those in non-inhibitor patients. The increased number of Treg cells and increased T-cell cytokines may be related to the development and efficiency of the factor VIII inhibitor.

Breeding new seedless grape by means of in vitro embryo rescue.

This project aimed at breeding new seedless grape cultivars by embryo rescue through three hybridization methods: 1) using cross-breeding between seedless Vitis vinifera cultivars and wild Chinese Vitis spp; 2) crossing with two seedless cultivars, and 3) hybridization between grapes of different ploidy. Genotype, sampling times, and media were confirmed to play important roles in this system. Among the different genotypes, the productions of hybrid plants were significantly different, ranging from 23.0% (Ruby Seedless x Black Olympia) to only 1.1% (Pink Seedless x Beichun), except for the combinations from which no surviving seedlings were obtained. We got the best sampling times, in days after flowering (DAF), from the following different combinations: 'Flame Seedless x Beichun' (39 DAF); 'Blush Seedless x Shuangyou' (54 DAF); 'Pink Seedless x Beichun' (54 DAF); 'DA7 x Shuangyou' (44 DAF); 'Blush Seedless x Thompson Seedless (54 DAF)'; 'Pink Seedless x Flame Seedless' (54 DAF); 'DA7 x Blush Seedless' (44 DAF); 'Ruby Seedless x Black Olympia' (63 DAF); 'DA7 x Jingyou' (44 DAF); 'Flame Seedless x Fujiminori' (39 DAF), and 'Big Black x Kyoho' (72 DAF). The highest rates of embryo formation (13.2%) and plant development (90.1%) were found when ovules were cultured in MM4 with 500 mg/L mashed banana. Conversely, they were reduced by addition of plant growth regulators. Seven new hybrids were successfully obtained. As a result of early nuclear-free character and ploidy level identification, 11 seedless grape lines, and 3 triploid and 2 haploid grape lines were obtained.

Activation of the ERK1/2 pathway by the CaMEK gene via adeno-associated virus serotype 9 in cardiomyocytes.

Extracellular signal-regulated kinase (ERK1/2) is one of the mitogen-activated protein kinases, key components of the reperfusion injury salvage kinase pathway, which plays an important role in protecting the myocardium from lethal ischemia-reperfusion injury. Constitutive activation of the mitogen-activated protein kinase kinase 1 (CaMEK) can promote ERK1/2 expression, which is thereby expected to exert protective action on the heart against ischemia-reperfusion injury. The adeno-associated virus serotype 9 vector (AVV9) is a novel tool for gene therapies targeting human diseases owing to its nonpathogenic capability for transducing nondividing cells and its long-term transgene expression. We used a recombinant AAV9 vector to deliver the CaMEK gene into cardiomyocytes and assessed whether AAV9 vector-mediated CaMEK gene transfection could enhance the long-term expression and activity of ERK1/2. Our observations suggest that AAV9-mediated gene expression is preferentially restricted to cardiomyocytes and that mediated CaMEK gene transfection enhanced the expression of ERK1/2 phosphorylation and consequently upregulated the expression of downstream components of ERK1/2 and its transcription factors.