A new species of the genus Psilota Meigen, 1822 (Diptera: Syrphidae) from China.

The genus Psilota Meigen, 1822 is recorded for the first time from China, and the species Psilota bashanensis Huo and Zhao sp. nov. is described and illustrated based on the adult male. The complete cytochrome c oxidase subunit I (COI) gene of this new species has been successfully obtained and compared to that of other congeneric species. An updated key to adult males of the genus Psilota from the Palaearctic Region is also provided.

A new species of the flower fly genus Criorhina Meigen (Diptera: Syrphidae) from mainland China.

The genus Criorhina consists of honey-bee and bumblebee mimic flower flies with a strongly produced face. It is widespread in the Holarctic and Oriental Regions. Criorhina adults are usually found flying near white spring flowers in woodlands and shrubs. The Chinese Criorhina fauna is poorly known and includes seven species. In our flower fly collection of Henan Province, northern China, an eighth new species was discovered: Criorhina rostrata Li, Huo Li sp. nov. This new species is here described and illustrated. The new species possess a very long proboscis, unique amongst the Criorhina species from mainland China. In addition, during the course of this study, Criorhina brevipila Loew, 1871 was also found to be present in mainland China, based on a specimen from our collection. A key to the species of Criorhina from mainland China is given.

Phytochemicals Content, Antioxidant and Antibacterial Activities of Sophora viciifolia.

The objective of this study is to compare the efficacy of ethanol extracts from different parts of Sophora viciifolia. The content of polyphenols, flavonoids, alkaloids, and antioxidant capacity, antimicrobial activity were investigated, and individual polyphenols and alkaloids were analyzed and quantified by ultra-high performance liquid chromatography (UPLC). The microdilution method was used to determine the antimicrobial activity of extracts from S. viciifolia on six strains. The results for extracts from the different parts (flowers, leaves, and fruit) were compared in varying concentrations to determine whether one extract source is superior to another. Testing verified that extracts from the different parts of S. viciifolia did vary, as expected. For example, extract from the leaves had the best antimicrobial activity against pathogenic Candida albicans, but all extracts had good antimicrobial activity against the six tested strains. These results reveal that the active substances in S. viciifolia are abundant and have good antioxidant and antimicrobial activities, which can provide theoretical support for the subsequent development and utilization of S. viciifolia extracts.

Microdon dentigiganteum sp. nov. and other Microdontinae species (Diptera: Syrphidae) from Northeast China.

Hoverflies (Diptera: Syrphidae) of the subfamily Microdontinae were surveyed in Northeast China. A total of six species were identified, including one new to science, Microdon dentigiganteum sp. nov. from Liaoning. This new species has a pair of very large-size posterior calcars on the scutellum and very wide flat tarsi. New records of Metadon spuribifasciatus (Huo, Ren et Zheng, 2007), Microdon analis (Macquart, 1842) and Microdon auricomus Coquillett, 1898 from Jilin province, and Microdon ignotus Violovitsh, 1976 and Microdon oitanus Shiraki, 1930 from Liaoning province are provided. A key to the six studied species and diagnostic figures are presented.

Spazigasteroides a new genus from China with a black face and scutellum in the Syrphini (Diptera: Syrphidae).

In the present paper, the genus Spazigasteroides gen. nov. (Diptera, Syrphidae), with Spazigasteroides caeruleus sp. nov. as type species, is described from China. The new genus bears the following characters: Head strongly concave posteriorly and closely appressed to thorax so that the bare postpronota are entirely hidden. Face black in ground colour. Antennae short, with basoflagellomeres slightly longer than wide. Scutellum black. Postmetacoxal bridge

SCYL1 binding protein 1 promotes the ubiquitin-dependent degradation of Pirh2 and has tumor-suppressive function in the development of hepatocellular carcinoma.

Pirh2 is a Ring-H2 domain containing E3 ubiquitin ligase that targets several important tumor suppressor genes for proteasomal degradation. Overexpression of Pirh2 is frequently detected in many clinical tumor tissues including hepatocellular carcinoma (HCC). However, the molecular mechanism of Pirh2 activation in tumorigenesis still remains poorly understood. In this study, we find a Pirh2-binding protein, SCYL1 binding protein 1 (SCYL1BP1), that can promote the ubiquitin-dependent degradation of Pirh2. SCYL1BP1 colocalized with Pirh2 in the cytoplasm and prevented its localization to the nucleus. Ectopic expression of SCYL1BP1 increased the expression of p53 and further inhibited the G(1)/S transition of HCC cell lines. Conversely, knock down of SCYL1BP1 restored the expression of Pirh2 and inhibited p53 at protein level. Functional assays found that reintroduction of SCYL1BP1 into HCC cell lines significantly inhibited cell proliferation, foci formation, colony formation in soft agar and tumor formation in nude mice, suggesting the strong tumor-suppressive function of SCYL1BP1 in HCC progression. Furthermore, SCYL1BP1 was found to be frequently downregulated in HCC clinical specimens compared to their paired non-tumor tissues by immunohistochemical staining. Taken together, our data suggested that the interaction of SCYL1BP1/Pirh2 could accelerate Pirh2 degradation through an ubiquitin-dependent pathway. SCYL1BP1 may function as an important tumor suppressor gene in HCC development.

Molecular analysis of Leonurus species in China based on ITS and matK sequences.

The genus Leonurus has long been recognized as a natural group, but its interspecific relationship has not yet been studied in the light of sequence data. The ITS regions and matK sequences of all subgenera of Leonurus in China were amplified, sequenced and investigated. Phylogenies generated by maximum parsimony and neighbor-joining methods and division of the genus into two major clades. The phylogenetic results indicated that L. chaituroides has the very close phylogenetic relationship with Subg. Cardiochilium and supported the notion that L. macranthus acts as the bridge between Subg. cardiochilium and Subg. Leonurus. According to the analysis of information given by ITS and matK sequences, we suggest that ITS sequences would be more suitable to serve as markers for authentication of Herba Leonuri than matK does.

Chromodomain helicase/adenosine triphosphatase DNA binding protein 1-like (CHD1l) gene suppresses the nucleus-to-mitochondria translocation of nur77 to sustain hepatocellular carcinoma cell survival.

UNLABELLED: Amplification of 1q21 has been detected in 58% to 78% of primary hepatocellular carcinoma cases, suggesting that one or more oncogenes within the amplicon play a critical role in the development of this disease. The chromodomain helicase/adenosine triphosphatase DNA binding protein 1-like gene (CHD1L) is a recently identified oncogene localized at 1q21. Our previous studies have demonstrated that CHD1L has strong tumorigenic ability and confers high susceptibility to spontaneous tumors in a CHD1L-transgenic mouse model. In this study, we demonstrate that the antiapoptotic ability of CHD1L is associated with its interaction with Nur77, a critical member of a p53-independent apoptotic pathway. As the first cellular protein identified to bind Nur77, CHD1L is able to inhibit the nucleus-to-mitochondria translocation of Nur77, which is the key step of Nur77-mediated apoptosis, resulting in the hindrance of the release of cytochrome c and the initiation of apoptosis. Knock-down of CHD1L expression by RNA interference could rescue the mitochondrial targeting of Nur77 and the subsequent apoptosis. Further studies found that the C-terminal Macro domain of CHD1L is responsible for the interaction with Nur77, and a CHD1L mutant lacking residues 600-897 failed to interact with Nur77 and prevented Nur77-mediated apoptosis. More importantly, we found that the inhibition of Nur77-mediated apoptosis by endogenous CHD1L is a critical biological cellular process in hepatocarcinogenesis. CONCLUSION: We demonstrate in this study that overexpression of CHD1L could sustain tumor cell survival by preventing Nur77-mediated apoptosis.

Identification of crude drugs from Chinese medicinal plants of the genus Bupleurum using ribosomal DNA ITS sequences.

To determine the variation of rDNA ITS sequences in medicinal plants of the genus Bupleurum and identify corresponding DNA molecular markers associated with medicinally important members, ITS regions were amplified, sequenced, and analyzed by DNAssist version 2.2. We found that the homologous alignment of ITS sequences between members of the genus Bupleurum and associated out-groups was lower than 75%, while within-group alignment was greater than 87%. The conclusion can be drawn that ITS sequences can be used as reliable molecular markers for the identification of Radix Bupleuri.

[Phylogeny relationship and molecular identification of ten Huperzia species (Huperziaceae) based on matK gene sequences].

OBJECTIVE: To study the phylogeny relationship and molecular identification of 10 species from Huperzia (Huperziaceae) based on matK gene sequences data. METHOD: Total DNA of nine species from Huperzia was extracted; matK gene sequence was amplified by PCR. PCR product was directly sequenced after purification. RESULT: The chloroplast matK gene nucleotide sequences from 9 species of Huperzia species were sequenced. The matK gene nucleotide sequences length was 1 589 bp. Analysis with Huperzia lucidula matK gene nucleotide sequences (download from GenBank) and taking Lycopodiella cernua as outgroup, Maximum Parsimony, Neighbor-Joining analyses and genetic distances were conducted using MEGA 3.1 software. 35 variable sites and 35 parsimony informative sites have been found. Pairwise genetic distances among 10 species of Huperzia was 1.59% - 0.25%. CONCLUSION: The results were consistent with the taxonomy in morphological of Huperzia. But H. longipetiolata and H. serrata were resolved into in different clade. There are 19 different sites of matK gene sequences between H. longipetiolata and H. serrata, the genetic distances is 0.121%. It is suggested H. longipetiolata should be as an independent species.

Analysis of genital Candida albicans infection by rapid microsatellite markers genotyping.

BACKGROUND: Candida albicans (C. albicans) infection, often occurring in genital candidiasis, has increased dramatically recently. Developing an efficient C. albicans typing method may contribute to understanding its epidemiological characteristics and guiding efficient treatment. We used rapid microsatellite genotyping assay for interstrain differentiation of C. albicans isolates and explored some characteristics of its spread. METHODS: DNA was extracted from C. albicans isolates from gentalia, recta and mouths of 39 female cases and 27 male cases of genital candidiasis. Three fluorescent primers for the microsatellite markers in conserved genes (CDC3, EF3 and HIS3) of C. albicans were used to amplify the isolates DNA by PCR. Fluorescent signals were read with an automatic sequencer and analyzed with GeneScan software. RESULTS: Analysis of the three microsatellites markers showed 18 gene allelic associations in genital C. albicans infected patients: 10 allelic associations in female and 11 allelic associations in male, of which 3 allelic associations shared by both genders covered 71% of infections. The most dominant allele association of pathogenic strains for both genders was 116:124, 122:131, 160:200 that covered about 50% of infection. Gentalia and recta shared the same strains in 80% of female patients, but in only 3.8% of male patients. There were 2.7% female patients, but no males, with same strain in both gentalia and mouths. Five of seven genital C. albicans infected couples had the same allelic associations of which 4 were the dominant pathogenic C. albicans susceptible for both genders. CONCLUSIONS: The predominant allelic association of the pathogenic strain in genital C. albicans infection is 116:124, 122:131, 160:200. Vaginal pathogenic strains are probably maintained from the rectal reservoir. Pathogenic strains of male patients are probably from frequent sexual intercourse. The aggressiveness of some strains varies with gender.

ITS sequence analysis used for molecular identification of the Bupleurum species from northwestern China.

Radix Bupleuri (Chaihu), a famous Traditional Chinese Medicine, is derived from the dried roots of Bupleurum chinense DC. and B. scorzonerifolium Willd. But nowadays, other nine species and varieties from genus Bupleurum are also used as Radix Bupleuri in northwestern China. The authentic identification of dried roots of B. chinense and B. scorzonerifolium, however, is difficult to just base on the appearance and morphology. A molecular genetic method was developed to help discriminate the original species of Radix Bupleuri. The ITS sequences ( approximately 600 bp) were amplified by the PCR technique from genomic DNA isolated from all the collected samples. According to analyzing the information given by ITS sequences, the conclusion can be made that ITS sequence could serve as markers for authentication of Radix Bupleuri.

Screen p53 mutations in hepatocellular carcinoma by FASAY: a novel splicing mutation.

OBJECTIVE: To establish a routine procedure for the detection of p53 mutations in hepatocellular carcinoma (HCC) surgical resections using the FASAY (functional analysis of separated alleles of p53 on yeast) procedure. METHODS: p53 status was analyzed by FASAY and cDNA sequencing in 50 cases of HCC. After the extraction of RNA from the frozen tumor and corresponding normal tissues, reverse transcription RT-PCR was carried out using these samples. The assay can detect mutations of p53 mRNA between codons 67 and 347 by the DNA-binding activity of the protein and reveal them as red colonies. RESULTS: Of the 50 specimens, 29 (58%) were positive (mutant) by FASAY. Sequencing analysis confirmed that all 29 FASAY positive tumors harbored mutations, and that no mutations were detectable in any FASAY negative tumors. In 29 p53 mutations, 22 mutations were point missense mutation, 5 were deletions and 2 were splicing mutations. A novel splice mutation on splice donor of intron 6 was reported, which could produce two different mRNAs, respectively using the nearest upstream and downstream recessive splice donor sites. CONCLUSION: FASAY is a sensitive method for detecting the various types of p53 mutations in HCC, suggesting that the yeast functional assay for the detection of p53 mutations may be essential for elucidating their clinical significance.

[Nuclear ribosomal DNA internal transcribed spacer 1 sequences of 4 Leonurus species].

OBJECTIVE: To analyze the nuclear ribosome DNA (nrDNA) internal transcribed spacer (ITS) sequences of 4 Leonurus species, and the possibility of using them for molecular authentication of the crude drugs from the genus. METHODS: The nrDNA ITS sequence (including ITS1, 5.8S rDNA, ITS2, and partial 18S rDNA and 26S rDNA) of L. japonicus and its 3 adulterant species were amplified and sequenced, and CLUSTRAL X and MEGA software was employed for analysis. RESULTS: The variation of ITS1 and ITS2 between L. japonicus and its adulterant species ranged between 7.2% and 18.8% and between 14.2% and 27%, respectively. The phylogenic tree derived from the dendrograms based on the ITS sequence data contained some discrepancy from the traditional classification. CONCLUSION: The nrDNA ITS sequences can be used potentially as efficient markers for identification of L. japonicus and its adulterants, and further study is needed for studying the phylogeny of Leonurus.

[ITS sequence of 9 Bupleurum species and its application in identification of Chaihu (Radix Buplurei)].

OBJECTIVE: The study the patterns of rDNA ITS sequence variation of 9 medicinal species of genus Bupleurum in China to find the DNA molecular markers for identification of these crude drugs. METHOD: The ITS regions of these species were amplified by PCR and sequenced, and their sequences were analyzed by DNAssist Version 2.2. RESULTS: Homologous alignment indicated that the consanguinity of the ITS sequences between genus Bupleurum and outgroups was lower than 75%, and that within genus Bupleurum was higher than 88%. For the same species, the consanguinity was higher than 99%. CONCLUSION: The ITS sequences may serve as reliable molecular markers for identification of Radix Bupleuri.

P5644 interacts with phosphatidylinositol-4-phosphate adaptor protein-1 associated protein-1.

The human novel gene pp5644 (GeneBank Accession No. AF289559) coding for 124 amino acids was recently cloned. Overexpression of pp5644 in Hela cells significantly inhibited the growth and colony formation. The pp5644-interacting protein FAPP1 (phosphatidylinositol-four-phosphate adaptor protein1) associated protein-1, called FASP1, was obtained by using yeast two-hybrid system. The interaction between pp5644 and FASP1 was experimentally confirmed by GST pull-down assay in vitro and co-immunoprecipitation assay in vivo. Co-localization of pp5644 and FASP1 in cytoplasm in Hela cells could further support the interaction. Based on the experimental results, it is suggested that pp5644 physically bind to FASP1 and the biological significance of this kind of interaction in vivo is discussed.

Identification of LZP gene from Mus musculus and Rattus norvegicus coding for a novel liver-specific ZP domain-containing secretory protein.

Zona pellucida (ZP) domain has been recognized in a number of receptor-like eukaryotic glycoproteins, which involved in many important biological processes, such as signal transduction, development, differentiation and so on. Here we report the identification of Mus musculus and Rattus norvegicus orthologues of Homo sapiens LZP gene which codes for a novel ZP domain-containing protein. Sequence analysis revealed that human, rat and mouse LZP proteins are highly conserved. Mouse LZP gene has two transcripts, 2.4 and 2.8 KB long respectively, coding for identical protein. Mouse LZP mRNA is expressed specifically in hepatocytes. Our data also showed that mouse LZP localizes mostly on nuclear envelope, and at the same time, it can be secreted into blood in a truncated form.

A novel liver-specific zona pellucida domain containing protein that is expressed rarely in hepatocellular carcinoma.

We currently identified a liver-specific gene that encodes a novel zona pellucida (ZP) domain-containing protein named liver-specific ZP domain-containing protein (LZP). The full-length complementary DNA (cDNA) of human LZP has 2,255 bp with a complete open reading frame (ORF) of 1,635 bp. The gene is localized on chromosome 10q21.3 and spans 40 kb with 9 encoding exons and 8 introns. The deduced protein sequence has 545 amino acid residues, with an N-terminal signal peptide followed by 3 epidermal growth factor (EGF)-like domains and a ZP domain in C-terminal section. Interestingly, human LZP is expressed specifically in liver out of 23 tissues examined, and its mouse counterpart was detected at very early stage during embryo development. Moreover, LZP can be secreted into blood, albeit the protein was localized mainly on the nuclear envelop of hepatocytes. Most importantly, LZP is down-regulated in hepatocellular carcinoma (HCC) and HCC cell lines; meanwhile, the decreased level of hLZP messenger RNA (mRNA) could, at least in some HCC samples, be related to the methylation status of the putative LZP promoter. However, overexpression of hLZP in HCC cell line SMMC-7721 and human liver cell line L02 by stable cell transfection did not inhibit cell growth, implying that the down-regulation of hLZP in HCC might be a consequence of the dedifferentiation involved in hepatocarcinogenesis. In conclusion, these data suggest that LZP is a liver-specific protein involved possibly in hepatocellular function and development, and the protein could be used as potential negative biomarker for HCC pathologic diagnosis.

Identification of a candidate oncogene SEI-1 within a minimal amplified region at 19q13.1 in ovarian cancer cell lines.

High-level amplification of DNA sequence at 19q13.1 is one of the frequent genetic alterations in ovarian cancer. In an attempt to verify the minimal amplified region (MAR) at 19q13.1 and to identify the target oncogenes, 49 probes within a region from D19S425 to D19S907 ( approximately 19.5 cM) were used to survey the amplification status in four ovarian cancer cell lines that have been confirmed as containing amplification at 19q13.1. Two separated overlapping MARs, MAR1 (approximately 200 kb) and MAR2 (approximately 1.1 Mb), were identified at 19q13.1. Two candidate oncogenes, AKT2 and SEI-1, were identified in MAR2. Amplification and overexpression of these two genes in four ovarian cancer cell lines were confirmed by Southern and Northern blot analyses. The proliferation-related function of AKT2 and SEI-1 suggests that both genes are likely to be biological targets of an amplification event at 19q13.1 in ovarian cancer and to play important roles in ovarian tumorigenesis.

Analysis of Phylogenetic Relationships Among Twelve Yeast Species.

Partial sequences of 25 S ribosomal DNA for nine Candida yeast and one Kluyveromyces yeast strains were cloned and determined. We compared them with two published 25 S rDNA sequences of Saccharomyces cerevisiae and Candida albicans. An evolutionary tree of the twelve species was inferred from about 370 sites of 5' end of 25 S ribosomal DNA using the methods of neighbor-joining and bootstrap. The molecular data indicate a very close affinity between Candida kefyr CBS834 and Kluyveromyces cicerisporus CBS4857.