Targeted gene therapy and in vivo bioluminescent imaging for monitoring postsurgical recurrence and metastasis in mouse models of liver cancer.

We investigated the effects of combined targeted gene therapy on recurrence and metastasis after liver cancer resection in nude mice. Twenty BALB/C mice were randomly divided into control and treatment groups with 10 mice in each group and a male/female ratio of 1:1. Luciferase gene-labeled human primary hepatic carcinoma cell line MHCC97-H was then used to prepare a carcinoma model. An optical in vivo imaging technique (OIIT) was used 10 days later to detect the distribution of tumor cells, followed by partial liver resection and gene therapy. In the treatment group, 100 mL phosphate-buffered saline (PBS) containing 1 x 1012 rAAV/AFP/IL-24 gene viral vectors was injected into liver sections and peritumoral posterior peritoneal tissues; in the control group, the same amount of PBS containing 1 x 1012 empty viral vectors was injected at the same sites. OIIT was then used to detect the in vivo tumor metastasis 21 days later. Luciferase gene-labeled human primary hepatic carcinoma cell line MHCC97-H successfully infected 20 nude mice, and OIIT showed that the two groups exhibited metastasis after local tumor resection, but there were more tumor cells in the control group (P < 0.05). rAAV/AFP/IL-24 gene therapy can inhibit recurrence after liver cancer resection.

Common genetic risk factors of venous thromboembolism in Western and Asian populations.

Venous thromboembolism (VTE) is a multifactorial disorder involving both acquired and genetic risk factors. The common genetic factors in Western populations have been studied and reported for several decades, while studies on Asian populations are relatively scarce. Evidence suggests that the prevalence and genetic risk factors of VTE vary significantly among ethnic populations. In this review, we summarize the common genetic risk factors of VTE in both Western and Asian populations. In addition to the development of DNA sequencing technology, genome-wide association studies have many advantages and are becoming more important in identifying new genetic risk factors and susceptible loci. They can therefore help in the prediction and prevention of VTE.

Phylogenetic analysis of Gansu sheeppox virus isolates based on P32, GPCR, and RPO30 genes.

Two outbreaks of sheeppox in sheep have occurred in Gansu Province, China. The P32, GPCR, and RPO30 genes were used as markers for differential diagnosis. We confirmed that the outbreaks were caused by sheeppox virus. Sequence and phylogenetic analysis of the P32, GPCR, and RPO30 genes revealed a close relationship between the 2 isolates and Chinese sheeppox viruses. Because ill sheep were imported from Jingyuan, another county of Gansu Province, our results strongly suggest the importance of veterinary surveillance prior to transportation.

A possible way that φC31 integrase regulates the recombination direction.

φC31 integrase encoded by Streptomyces phage can mediate site-specific recombination between phage and host genomes. The recombination direction is generally considered to be accurately regulated, but the regulatory mechanisms involved are still unclear. Recently, some hyperactive mutants of φC31 integrase that can bypass the regulatory steps have been isolated and extensively studied. A putative coiled-coil region is found to play a critical role in controlling recombination direction. Further analysis led us to the speculation that at least two regions in the N-terminal domain of φC31 integrase are involved in the tetrameric interfaces and that the putative coiled coil interacts with one of the regions to regulate the recombination direction.

Development of simple sequence repeat markers in persimmon (Diospyros L.) and their potential use in related species.

Persimmon (Diospyros L.) is an economically important fruit in the world, and it has been recognized as a healthy nutrient supply for human consumption. In this study, 14 microsatellite markers were developed from an AG/TC and AC/TG-enriched genomic library of Chinese persimmon Mopanshi. Twelve polymorphic markers were selected in 4 related species; these markers showed transferability to the 4 related persimmon species. In addition, 10 simple sequence repeat (SSR) markers were used to detect the genetic diversity among 51 persimmon accessions from China, Japan, and Korea. A total of 57 polymorphic bands with an average of 5.7 bands per primer pair were observed. According to cluster analysis and principal coordinate analysis, all persimmon accessions could be divided into 4 groups. A close relationship existed between D. kaki and D. oleifera, and D. glaucifolia and D. lotus. Jinzaoshi could be considered a separate species of persimmon. These new SSR markers provide tools for evaluating genetic relatedness among different persimmon species.

Genetic diversity and population structure of Stipa bungeana, an endemic species in Loess Plateau of China, revealed using combined ISSR and SRAP markers.

Sequence-related amplified polymorphism (SRAP) and inter-simple sequence repeat (ISSR) markers were used to assess the genetic diversity within and among 15 natural populations of Stipa bungeana from the Loess Plateau of China. Using 15 SRAP primers, 504 (99.80%) polymorphic loci were detected, and 372 polymorphic loci (96.12%) were identified using 15 ISSR primers. At the species level, the S. bungeana populations showed relatively low levels of genetic diversity (HE = 0.2017 for SRAP; HE = 0.2066 for ISSR). The results of analysis of molecular variance indicated that genetic variation within populations (42.02% for SRAP and 38.40% for ISSR) is lower than that among populations (57.98% for SRAP and 61.60% for ISSR). The genetic distance was significantly correlated with geographical distance by the Mantel test (r = 0.3978, P = 0.002). Our results demonstrated that both SRAP and ISSR markers are effective and reliable for assessing the genetic diversity of S. bungeana. In addition, these data inform conservation and breeding strategies for S. bungeana.

Association between single nucleotide polymorphisms of the osteoprotegerin gene and postmenopausal osteoporosis in Chinese women.

Osteoporosis is an important and common complex health problem, particularly in postmenopausal women. It is characterized by a reduction in bone mineral density (BMD) and a deterioration of bone microarchitecture with a consequent increase of fracture risk. The osteoprotegerin (OPG) gene is considered to play an important role in the pathogenesis of osteoporosis. We analyzed SNPs of the OPG gene and associations between these polymorphisms and BMD in 399 Chinese postmenopausal women. BMD was quantified at the lumbar spine (L2-4), femoral neck, and total hip. The g.2264T>C and g.27676A>C SNPs were detected by PCR-RFLP and DNA sequencing methods. A significant association with spine BMD was found for g.27676A>C. The spine BMD value for subjects with genotype AA was significantly higher than those with genotypes GA and AA. No significant association was detected between any of the SNP marker genotypes and the other traits. We conclude that g.27676A>C in the OPG gene affects spine BMD and that the C allele is associated with increased risk for osteoporosis in Chinese postmenopausal women.

Analysis of genetic diversity among Chinese wild Vitis species revealed with SSR and SRAP markers.

The genetic diversity among 80 Vitis materials including 62 indigenous accessions of 17 wild Vitis species in China and 7 interspecific hybrids, 10 V. vinifera L. cultivars, and 1 V. riparia Michaux were evaluated by simple sequence repeat and sequence-related amplified polymorphism markers. A total of 10 simple sequence repeat primers and 11 sequence-related amplified polymorphism primer combinations were amplified, and 260 bands were generated, of which 252 were polymorphic with an average polymorphism rate of 97.02%. Genetic relationships among the different Vitis species indicated that V. ficifolia and V. yeshanensis could be considered a separate species. As for the 4 major ecogeographic regions of Chinese wild Vitis species, the genetic diversities of Chinese wild Vitis species from the Qinling Mountain region (H = 0.1947, I = 0.3067) and the mid-downstream Yangtze River region (H = 0.1834, I = 0.2925) were higher, with results suggesting that these regions may be one of the major centers of Vitis origin. An understanding of the genetic diversity of these Chinese wild Vitis species could provide the theoretical foundation for further protection and reasonable utilization in grape breeding.

Genome-wide identification, characterization, and expression analysis of the MLO gene family in Cucumis sativus.

Mildew resistance locus o (MLO) is a plant-specific seven-transmembrane (TM) gene family. Several studies have revealed that certain members of the MLO gene family mediate powdery mildew susceptibility in three plant species, namely, Arabidopsis, barley, and tomato. The sequenced cucumber genome provides an opportunity to conduct a comprehensive overview of the MLO gene family. Fourteen genes (designated CsMLO01 through CsMLO14) have been identified within the Cucumis sativus genome by using an in silico cloning method with the MLO amino acid sequences of Arabidopsis thaliana and rice as probes. Sequence alignment revealed that numerous features of the gene family, such as TMs, a calmodulin-binding domain, peptide domains I and II, and 30 important amino acid residues for MLO function, are well conserved. Phylogenetic analysis of the MLO genes from cucumber and other plant species reveals seven different clades (I through VII). Three of these clades comprised MLO genes from A. thaliana, rice, maize, and cucumber, suggesting that these genes may have evolved after the divergence of monocots and dicots. In silico mapping showed that these CsMLOs were located on chromosomes 1, 2, 3, 4, 5, and 6 without any obvious clustering, except CsMLO01. To our knowledge, this paper is the first comprehensive report on MLO genes in C. sativus. These findings will facilitate the functional characterization of the MLOs related to powdery mildew susceptibility and assist in the development of disease resistance in cucumber.