Chang Wu

The complete mitochondrial genome of the flesh fly, Boettcherisca peregrine (Diptera: Sarcophagidae)

Abstract The complete mitochondrial genome of Boettcherisca peregrine (B. peregrina), an important forensic entomology, was sequenced for the first time. The 14,922 bp circular genome contains 37 genes that were found in a typical Metazoan genome: 13 protein-coding genes, 22 transfer RNA genes and 2 ribosomal RNA genes. It also contains one non-coding A + T-rich region. The arrangement of the genes was the same as that found in the other insect. The overall base composition on heavy strand was as follows: A, 38.86%; G, 15.10%; C, 9.93%; T, 36.11%; and the A + T content 74.97%. The mitochondrial genome of Sarcophaga presented could be valuable for resolving phylogenetic relationships within the order Diptera and especially for the family Sarcophagidae. The molecular data presented may also be used to screen favorable molecular markers for species identifications for forensic entomology purposes.

The complete mitochondrial genome of the scuttle fly, Megaselia scalaris (Diptera: Phoridae)

Abstract More than 1400 scuttle flies species in worldwide comprise the Megaselia genus, the largest genus in the family Phoridae. The complete mitochondrial genome of Megaselia scalaris, a medically important entomology was sequenced for the first time. The 15,599 bp circular genome contains the 37 genes found in a typical Metazoan genome: 13 protein-coding genes, 22 transfer RNA genes and 2 ribosomal RNA genes. The mitochondrial genome also contains one non-coding A + T-rich region. The arrangement of the genes was identical with other insect. Each of the base composition on heavy strand was as follows A: 38.87%, G: 13.74%, C: 9.46%, T: 37.93% and the A + T content 76.80%. The mitochondrial genome of M. scalaris presented may be valuable for determining phylogenetic relationships within the order Diptera and especially for the family Phoridae. These sequences could also be used to select reliable molecular markers for species identification in forensic entomology.

Expression of C-X-C chemokine receptor types 1/2 in patients with gastric carcinoma: Clinicopathological correlations and significance

C-X-C chemokine receptor types 1/2 (CXCR1/2) may play multiple roles in the development and progression of a number of types of tumor. The abnormal expression of CXCR1/2 in various types of malignant tumors has been reported, but less is known with regard to gastric carcinoma. The present study was preliminarily conducted to elucidate the correlation between clinicopathological factors and the immunohistochemical expression of CXCR1/2 in patients with gastric carcinoma. The expression of CXCR1/2 in 69 specimens of sporadic gastric carcinoma and their corresponding non-neoplastic mucosa obtained by gastrectomy was assayed by immunohistochemistry (IHC) using a polyclonal anti-CXCR1/2 antibody. ERK1/2 and AKT phosphorylation and the expression of indicators of proliferation, growth and apoptosis (Bcl-2 and Bax, Cyclin D1, EGFR and Ki-67), angiogenesis (VEGF and CD34), invasion and metastasis (MMP-9, MMP-2, TIMP-2 and E-cadherin) were also detected by IHC. A total of 68 (98.6%) of the 69 patients with gastric carcinoma were found to have positive CXCR1/2 expression, which appeared to be significantly higher in gastric carcinoma compared with corresponding non-neoplastic mucosa tissues. The expression of CXCR1/2 in gastric carcinoma was significantly associated with invasion, metastasis and TNM staging (P<0.001). Correlation analysis between CXCR1/2 and pAKT (P=0.032), pERK (P<0.001), Cyclin D1 (P=0.049), EGFR (P=0.013), Bcl-2 (P=0.003), microvessel density (P=0.001), MMP-9 (P=0.013) and MMP-2 (P=0.027) expression using the Spearman test showed significant correlation in gastric carcinoma. Univariate and multivariate logistic regression analysis showed that, compared with negative or weak expression, overexpression of CXCR1/2 protein was a significant risk factor for TNM stage (P<0.001). These results preliminarily suggest that CXCR1/2 may be a useful maker for progression of the tumors and a promising target for gastric carcinoma therapy.

Selection of reference genes for quantitative gene expression studies in the house fly (Musca domestica L.) using reverse transcription quantitative real-time PCR

Ming Zhong1, Xiang Wang1, Jifang Wen1*, Jifeng Cai2, Chang Wu1, and Sanaa Mohamed Aly2 Department of Pathology, School of Basic Medical Sciences, Central South University, Changsha 410013, China Department of Forensic Medicine, School of Basic Medical Sciences, Central South University, Changsha 410013, China *Correspondence address. Tel: þ86-731-82650400; Fax: þ86-731-82650410; E-mail: jifangw@yahoo.com

Molecular characterization of the carbon dioxide receptor in the oriental latrine fly, Chrysomya megacephala (Diptera: Calliphoridae)

The blowfly Chrysomya megacephala (Fabricius) (Diptera: Calliphoridae) cannot only act as a mechanical vector of various pathogens, but also infest man and animals causing human health problems and economic losses in the livestock and fish industries. As in other insects, olfaction of this species plays an important role in host location and is presumably mediated via transmembrane receptor signaling pathways. Here, we isolate and characterize CmegGr1 and CmegGr2, two new members of the chemosensory receptor gene family from C. megacephala. The open reading frames of CmegGr1 and CmegGr2 cDNA clones encode 453 and 486 amino acid residues, respectively. These two deduced proteins display high amino acid conservation with previously identified carbon dioxide (CO2) receptors, such as Drosophila melanogaster Gr21a/Gr63a and Anopheles gambiae s.s. Gr22/Gr24. Further sequence analysis showed that both proteins are consistent with their corresponding orthologs in the membrane topology prediction with some ambiguities in the location of N terminus and the number of transmembrane domains. The transcripts of CmegGr1 and CmegGr2 were detected in the major chemosensory organs including the antennae and proboscises with maxillary palps attached. These results suggest that CmegGr1 and CmegGr2 are likely to be the primary receptors for CO2 detection in C. megacephala. Knowledge of the molecular identity of the blowfly olfactory CO2 receptors may aid in the development of novel control strategies designed to take advantage of this unique and critical olfactory pathway.

Proteomics-based analysis of differentially expressed proteins in the CXCR1-knockdown gastric carcinoma MKN45 cell line and its parental cell

C-X-C chemokine receptor types 1 (CXCR1), a cell-surface G-protein-coupled receptor has been found to be associated with tumorigenesis, development, and progression of some tumors. Previously, we have found that CXCR1 overexpression is associated with late-stage gastric adenocarcinoma. We also have demonstrated that knockdown of CXCR1 could inhibit cell proliferation in vitro and in vivo. In this study, we compared the changes of protein expression profile between gastric carcinoma MKN45 cell line and CXCR1-knockdown MKN45 cell line by 2D electrophoresis. Among the 101 quantified proteins, 29 spots were significantly different, among which 13 were down-regulated and 16 were up-regulated after CXCR1 knockdown. These proteins were further identified by mass spectrometry analysis. Among them, several up-regulated proteins such as hCG2020155, Keratin8, heterogeneous nuclear ribonucleoprotein C (C1/C2), and several down-regulated proteins such as Sorcin, heat shock protein 27, serpin B6 isoform b, and heterogeneous nuclear ribonucleoprotein K were confirmed. These proteins are related to cell cycle, the transcription regulation, cell adherence, cellular metabolism, drug resistance, and so on. These results provide an additional support to the hypothesis that CXCR1 might play an important role in proliferation, invasion, metastasis, and prognosis, and drug resistance of gastric carcinoma.

Evaluation of potential reference genes for qRT-PCR studies in human hepatoma cell lines treated with TNF-α

In this study, the expression of eight candidate reference genes, B2M, ACTB, GAPDH, HMBS, HPRT1, TBP, UBC, and YWHAZ, was examined to identify optimal reference genes by quantitative reverse transcription-polymerase chain reaction (qRT-PCR) analysis in two human hepatoma cell lines, BEL-7402 and SMMC-7721, treated with tumor necrosis factor-α (TNF-α) for different time periods. The expression stability of these genes was analyzed by three independent algorithms: geNorm, NormFinder, and BestKeeper. Results showed that TBP was the most stably expressed gene in BEL-7402 and SMMC-7721 cell lines under current experimental conditions, and that the optimal set of reference genes required for accurate normalization was TBP and HMBS, based on the pairwise variation value determined with geNorm. UBC and ACTB were ranked as the least stable genes by same algorithms. Our findings provide evidence that using TBP alone or in combination with HMBS as endogenous controls could be a reliable method for normalizing qRT-PCR data in human hepatoma cell lines treated with TNF-α.

Expression of miR-224, miR-145, and their putative target ADAM17 in hepatocellular carcinoma

Yuwu Liu1,2,3, Chang Wu4, Ying Wang1,2, Sailan Wen1,2, Junpu Wang1,2, Zhihong Chen1,2, Qiongqiong He1,2, and Deyun Feng1,2* Department of Pathology, Xiangya Hospital, Central South University, Changsha 410008, China Department of Pathology, School of Basic Medical Sciences, Central South University, Changsha 410013, China Department of Morphology, The Institute of Advanced Occupation Technology, Xinjiang Medical University, Urumqi 830011, China Department of Pathology, Shenzhen Sixth People’s Hospital (Nanshan Hospital), Shenzhen 518052, China *Correspondence address: Tel/Fax: þ86-731-82650410; E-mail: DeyunFeng2014@126.com

Loss of plexin-B3 in hepatocellular carcinoma.

Plexins are the primary receptors of semaphorins, and participate in the majority of intracellular pathways triggered by semaphorins, including the regulation of cell adhesion and the motility of numerous cell types. Recently, several studies have reported that plexins can significantly affect different aspects of cancer cell biology, and the aberrant expression of plexins has been observed in a wide variety of tumor types. However, the expression and role of plexin-B3 in hepatocellular carcinoma (HCC) is yet to be investigated. In the present study, plexin-B3 expression was measured in 14 paired HCC samples and the corresponding adjacent non-cancerous tissue by quantitative polymerase chain reaction and western blot analysis. The results indicated that the mRNA and protein expression levels of plexin-B3 were downregulated in HCC samples when compared with the corresponding adjacent non-cancerous tissue. In order to elucidate the correlation between clinicopathological data and the expression of plexin-B3 in patients with HCC, 84 HCC archived specimens were analyzed by immunohistochemistry (IHC). The IHC results revealed that the protein expression level of plexin-B3 was lower in the HCC samples compared with the corresponding adjacent non-cancerous tissue, and plexin-B3 underexpression was correlated with the patient gender and tumor size. In conclusion, these results indicated that loss of plexin-B3 in HCC may be of predictive value for the occurrence and progression of HCC. Thus, plexin-B3 may be a promising biomarker for the diagnosis and treatment of tumors in the future.