Significantly upregulated TACSTD2 and Cyclin D1 correlate with poor prognosis of invasive ductal breast cancer.

The tumor-associated calcium signal transducer 2 (TACSTD2) gene has been reported to be highly expressed in many types of human epithelial cancers, and is associated with tumor metastasis and poor prognosis. The aims of the present investigation were to analyze the TACSTD2 and Cyclin D1 expression at the mRNA and protein levels and to assess its prognostic significance in invasive ductal breast cancer (IDC). The expressions of TACSTD2 and Cyclin D1 in IDC tissues were consistently higher than those in the tumor-adjacent non-malignant tissues by a one-step real-time polymerase chain reaction and immunohistochemistry (P<0.001 and P=0.023, respectively). The statistical analysis of clinicopathologic characteristics and immunohistochemistry by the χ(2) test showed that the high expression of TACSTD2 in IDC was correlated to histological grade (P=0.023), P53 status (P=0.042), Cyclin D1 status (P<0.001), lymph node metastasis (P<0.001), distant metastasis (P=0.004) and TNM staging (P<0.001). Kaplan-Meier survival and Cox regression analyses were performed to evaluate the prognosis of IDC. These analyses also showed that a high TACSTD2 expression (P=0.003), a high Cyclin D1 expression (P=0.041), and lymph node metastasis (P=0.006) were independent prognosis factors. Collectively, our studies demonstrated that the high expression of TACSTD2 correlates with a poor prognosis in IDC.

First report on co-occurrence knockdown resistance mutations and susceptibility to beta-cypermethrin in Anopheles sinensis from Jiangsu Province, China.

The increasing prevalence of insecticide resistance in Anopheles sinensis, a major vector of malaria in Jiangsu province in eastern China, threatens to compromise the successful use of insecticides in malaria control strategies. It is therefore vital to understand the insecticide resistance status of An. sinensis in the region. This study examined the nucleotide diversity of the para-sodium channel and knockdown resistance (kdr) in five field populations of adult An. sinensis mosquitoes collected in Jiangsu province, identifying the L1014F and L1014C substitutions for the first time. Competitive polymerase chain reaction (PCR) amplification of specific allele (cPASA) and polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) for resistance diagnosis were developed and validated. Comparing the results with direct sequencing revealed that the PCR-RFLP method was more sensitive and specific whereas the cPASA method was more convenient and suitable. The significant positive correlation between kdr allele frequency and bioassay-based resistance phenotype demonstrates that the frequency of L1014F and L1014C substitutions in the kdr gene provides a useful molecular marker for monitoring beta-cypermethrin resistance in natural populations of An. sinensis. Our results point to the L1014F substitution as the key mutation associated with beta-cypermethrin resistance. The high resistance and mutation frequency detected in the five populations also suggest cross-resistance with other pyrethroids may occur in An. sinensis, highlighting the need for further surveys to map insecticide resistance in China and the adoption of a rational management of insecticide application for resistance management and mosquito vector control.

[Expression and clinical significance of Trop-2 in human pancreatic cancer].

OBJECTIVE: To investigate the expression and its clinical significance of Trop-2 in human pancreatic cancer. METHODS: Reverse transcription-polymerase chain reaction (RT-PCR) and immunofluorescence assay were used to characterize the expression of Trop-2 in human pancreatic cancer. The expression of Trop-2 protein in 31 tumor tissue samples, including peritumoral tissue from patients with pancreatic cancer, was detected with immunohistochemistry by tissue microarray. The clinicopathological characteristics and the tissue expression of Trop-2 protein were analyzed statistically. RESULTS: RT-PCR, immunofluorescence assay and immunohistochemistry by tissue microarray showed a high expression of Trop-2 in pancreatic cancer. The expression rate of Trop-2 was much higher in pancreatic cancer than that in peritumoral tissues (87.1% vs 9.7%). And a high expression of Trop-2 in pancreatic cancer was correlated with the low-differentiated changes. It had statistical significance (P < 0.05). In contrast, no statistically significant correlation was found between the expression of Trop-2 and gender or age. CONCLUSION: The expression of Trop-2 is correlated with the development and malignancy of pancreatic cancer. As a clinical prognostic marker, Trop-2 may be a potential therapeutic target for pancreatic cancer.

Generation and characterization of the human neutralizing antibody fragment Fab091 against rabies virus.

AIM: To transform the human anti-rabies virus glycoprotein (anti-RABVG) single-chain variable fragment (scFv) into a Fab fragment and to analyze its immunological activity. METHODS: The Fab gene was amplified using overlap PCR and inserted into the vector pComb3XSS. The recombinant vector was then transformed into E coli Top10F' for expression and purification. The purified Fab was characterized using SDS-PAGE, Western blotting, indirect ELISA, competitive ELISA, and the fluorescent antibody virus neutralization test (FAVN), respectively, and examined in a Kunming mouse challenge model in vivo. RESULTS: A recombinant vector was constructed. The Fab was expressed in soluble form in E coli Top10F'. Specific binding of the Fab to rabies virus was confirmed by indirect ELISA and immunoprecipitation (IP). The neutralizing antibody titer of Fab was 10.26 IU/mL. The mouse group treated with both vaccine and human rabies immunoglobulin (HRIG)/Fab091 (32 IU/kg) showed protection against rabies, compared with the control group (P<0.05, Logrank test). CONCLUSION: The antibody fragment Fab was shown to be a neutralizing antibody against RABVG. It can be used together with other monoclonal antibodies for post-exposure prophylaxis of rabies virus in future studies.

[Influence of F protein of hepatitis C virus subtype 1b inhibits on human hepatocellular carcinoma HepG2 cell apoptosis].

OBJECTIVE: To investigate the effects of F protein of hepatitis C virus subtype 1b on the apoptosis of human hepatocellular carcinoma HepG2 cells. METHODS: HepG2 cells were transfected with recombinant plasmid pcDNA3.0-F-EGFP and pcDNA3.0-F-EGFP-HepG2 strain was exposed to Act-D and tumor necrosis factor alpha (TNF alpha) treatment in order to induce cell apoptosis with positive control pcDNA3.0-C-EGFP-HepG2, negative control pcDNA3.0-C-EGFP-HepG2 and blank control HepG2. Annexin V-FITC/PI of flow cytometry was performed to determine the number of apoptotic cells. DNA Ladder was used to observe the isolation of apoptotic DNA fragments in the apoptotic cells. RESULTS: pcDNA3.0-F-EGFP- HepG2 cell strain showed a much delayed apoptosis as well as obviously lowering the apoptotic rate when compared with the pcDNA3.0-HepG2 strain and HepG2 strain (P < 0.001). CONCLUSION: The introduction and expression of extraneous gene (the F gene of hepatitis C virus subtype 1b) could significantly inhibit the apoptosis of HepG2 cells.

[Mass spectrometric analysis of peripheral blood mononuclear cells in HCV infected patients].

AIM: To investigate the diversity of proteins' pattern of the peripheral blood mononuclear cells (PBMCs) in patients with hepatitis C virus (HCV) infection, identify the differentially expressed proteins and analyze their roles in the mechanism of chronic hepatitis C. METHODS: The total proteins from PBMCs in HCV infected patients (28) and healthy persons (10) were separated by the immobilized pH gradient-based 2-DE. The differentially expressed proteins were screened by PDQuest analysis software, and then identified by peptide mass fingerprint (PMF) based on matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) and database searching. RESULTS: 2-DE results showed that the mean number of protein spots in HCV infected patients and healthy persons was 625 and 614. Twelve differential proteins were tested by MALDI-TOF-MS, and 10 of them were identified. Some of the proteins participated in protein synthesis and degradation, signal transduction, metabolism, or cytoskeleton construction while some of them were proteins of virus. CONCLUSION: 2-DE pattern of PBMCs from HCV infected patients or healthy persons has been established and ten differentially expressed proteins has been characterized. Our study is helpful for clinical detection of HCV infection and further research into the mechanisms of chronic hepatitis C.

[Molecular epidemiological study on the host and role of the Hantavirus and Orientia tsutsugamushi in the same epidemic area].

OBJECTIVE: To investigate whether Hantavirus (HV) and Orientia tsutsugamushi ( OT) can naturally infect and coexist in their host and role. METHODS: By field epidemiological study, Leptotrombidium scutellare (3829) was collected and separated from mice(166) in epidemic areas. The cells of mites separated from their host and role were cultured. PCR was used to detect HV-RNA and OT-DNA in the cell culture. RESULTS: In 105 Apodemus agrarius, 3 HV-RNA positive, 2 OT-DNA positive and 2 coinfection with HV and OT were detected;in 41 Brown rattus, 2 HV-RNA positive, 1 OT-DNA positive and 1 co-infection with HV and OT were detected. From 15 mites co-infected with HV and OT, 2 strains of HV pathogen, 2 strains of OT pathogen were separated and 1 HV and OT pathogen in the same mite were separate. CONCLUSION: The study demonstrates that co-infection of HV and OT did simultaneously exist in wild Leptotrombidium scutellare. This theory has some significance to the epidemic and precaution of HV and OT.

[Detection the coinfection of Hantavirus and Orientia tsutsugamushi in primary cultured mite cells].

OBJECTIVE: To investigate whether HV and Ot can coexist in their host (Leptotrombidium scutellare). METHODS: Collecting the separate Leptotrombidium scutellare and the ones from mice in epidemic area. The cells of mites at larva, nymph, and adult stages were cultured and made into smear. In situ RT-PCR and PCR were used to detect and locate HV RNA and Ot DNA in the primary cultured cells. RESULTS: Positive signals of HV RNA and Ot DNA distributed mostly in epithelial cells of digestive system and ovary cells of larva and nymph. The positive rate increased by the generation of passages. CONCLUSION: Coinfection of HV and Ot did exist in wild Leptotrombidium scutellare.

[Detection of serum anti-F antibody in hepatitis C virus infected patients].

OBJECTIVE: To assess the prevalence of serum anti-F in patients with hepatitis C virus (HCV) infection and the distribution of anti-F. METHODS: The recombinant protein (HCV-F/GST) was coated onto micro titer plates as antigen. Sera of 120 patients with hepatitis C virus infection, 15 patients with hepatitis B, 3 patients with hepatitis E and 10 normal sera were tested by indirect ELISA for detecting anti-F. RESULTS: 82 samples out of the 120 (68%) HCV infected patients exhibited a positive anti-F reaction, showing significant difference from the controls with no HCV infection (P < 0.01). Data from logistic analysis showed that the positive rate of anti-F was higher in patients over 50 year olds (OR = 6.675, 95% CI: 2.407-19.071). Patients of midrange, severe phase and hepatic cirrhosis had higher rate than the others (OR = 2.749, 95% CI: 1.470-5.141). CONCLUSION: Prevalence and distribution of anti-F in Yixing hepatitis C patients was reported and which might be related to the progression of HCV infection.