A study of effect of lncRNA MVIH on sensitivity of gastric cancer cells to gemcitabine.

OBJECTIVE: To study the effect of expression of long non-coding ribonucleic acid (lncRNA) associated with microvascular invasion in hepatocellular carcinoma (MVIH) on the sensitivity of gastric cancer cells to gemcitabine. MATERIALS AND METHODS: BGC-823 cell lines were divided into control group (no treatment), low expression group (lentiviral transfection with sh-lncRNA MVIH), and high expression group (lentiviral transfection with lncRNA MVIH). The expression of lncRNA MVIH, the protein expressions of E-cadherin and Vimentin, and the differences in proliferation, migration, invasion, and apoptosis of gastric cancer cells were detected via Reverse Transcription-Polymerase Chain Reaction (RT-PCR), Western blotting, Cell counting kit-8 (CCK-8) assay, transwell assay, wound healing assay, methyl thiazolyl tetrazolium (MTT) assay, and flow cytometry. RESULTS: The results of RT-PCR revealed that compared with that in control group, the mRNA expression of lncRNA MVIH was significantly decreased in low expression group and markedly increased in high expression group. Also, there were statistically significant differences (p<0.05). The results of Western blotting showed that compared with those in control group and low expression group, the protein expression of E-cadherin was significantly decreased, while the protein expression of Vimentin was markedly increased in high expression group (p<0.05). The results of transwell assay manifested that the number of invading gastric cancer cells was the largest in high expression group at 48 h (p<0.05), significantly larger than that in control group and low expression group (p<0.05), while it was the smaller in low expression group. It was found through the wound healing assay that the migration ability of gastric cancer cells was enhanced in high expression group, markedly stronger than in control group and low expression group, while it significantly declined in low expression group compared to control group. Besides, the results of CCK-8 assay showed that compared with that in control group, the sensitivity of gastric cancer cells to gemcitabine was remarkably increased in low expression group (p<0.05), while it significantly declined in high expression group (p<0.05). Finally, according to the flow cytometry, the apoptosis rate of gastric cancer cells was markedly higher in low expression group than that in control group and high expression group (p<0.05), indicating that the low expression of lncRNA MVIH can promote the apoptosis of gastric cancer cells. CONCLUSIONS: Reducing the expression of lncRNA MVIH can significantly lower the migration and invasion ability of gastric cancer cells and raise the sensitivity of gastric cancer cells to gemcitabine.

ALOX5AP rs10507391 polymorphism and the risk of ischemic stroke in Caucasians: an update meta-analysis.

Some reports evaluated the association between ALOX5AP rs10507391 polymorphism and the risk of ischemic stroke in Caucasians. The results remained unknown. Thus, we did a meta-analysis to evaluate this association. Nine case-control studies with 4198 patients and 3699 controls were included in this meta-analysis. A significant association was found between ALOX5AP rs10507391 polymorphism and ischemic stroke risk in Caucasians (OR=1.18; 95%CI, 1.08-1.28; P=0.0002). ALOX5AP rs10507391 polymorphism was associated with ischemic stroke risk in Caucasians from Europe (OR=1.20; 95%CI, 1.09-1.32; P=0.0002) but not from other countries (OR=1.13; 95%CI, 0.95-1.36; P=0.17). No significant association was found between ALOX5AP rs10507391 polymorphism and ischemic stroke risk in males (OR=1.12; 95%CI, 0.91-1.39; P=0.28). Moreover, ALOX5AP rs10507391 polymorphism was not associated with cardioembolic ischemic stroke risk (OR=1.04; 95%CI, 0.73-1.48; P=0.84). In conclusion, this study found that ALOX5AP rs10507391 polymorphism was associated with ischemic stroke risk in Caucasians.

Characterization of the complete mitochondrial genome of Setaria digitata (Nematoda: Setariidae) from China.

Setaria digitata is a filarial parasite that causes fatal cerebrospinal nematodiasis in goats, horses and sheep, resulting in substantial economic losses to livestock farmers. In the present study, the complete mitochondrial (mt) genome of S. digitata from China was determined, characterized and compared with that of S. digitata from Sri Lanka. The identity of the mt genomes was 98.3% between S. digitata from China and Sri Lanka, and the complete mt genome sequence of S. digitata from China was slightly shorter (25 bp) than that from Sri Lanka. For the 12 protein genes, this comparison revealed sequence differences at both the nucleotide (1.4%) and amino acid (2.2%) levels. The present study determined the complete mt genome sequence of S. digitata from China, providing novel genetic markers for the study of the population genetics and molecular epidemiology of S. digitata in animals.

Evidence for a role of GPRC6A in prostate cancer metastasis based on case-control and in vitro analyses.

OBJECTIVE: G protein-coupled receptor, family C, group 6, member A, (GPRC6A) is a prostate cancer (PCa) susceptibility gene and has been shown to regulate PCa progression. However, its role in PCa metastasis is largely unknown. The aim of this study was to confirm the association between GPRC6A and aggressive PCa in a case-control analysis, and to explore the function of GPRC6A in PCa metastasis in vitro. PATIENTS AND METHODS: The association of 14 single nucleotide polymorphisms (SNPs) of GPRC6A and linked to GPRC6A were evaluated with PCa risk and aggressive PCa in 916 subjects. Metastasis behavior was determined in GPRC6A knockdown PC3 cells, and the expressions of matrix metalloproteinase (MMP)2 and MMP9 were detected. Bone transcription factor runt-related transcription factor 2 (RUNX2) and epithelial-mesenchymal transition (EMT) marker genes were examined in the GPRC6A overexpression PC3 cells. RESULTS: Among the 14 SNPs tested in PCa patients and controls, 4 were associated with aggressive PCa (p = 0.032-0.037, odds ratio = 1.38-1.41). Both the migration and invasion abilities were reduced in PC3 cells that were transiently transfected with GPRC6A short interfering RNA (siRNA). The GPRC6A knockdown cells showed reduced activity levels of MMP2 and MMP9. Furthermore, RUNX2, EMT and ERK signaling were shown to be up-regulated in GPRC6A overexpression cells. CONCLUSIONS: These findings suggest that GPRC6A is associated with aggressive PCa. GPRC6A knockdown inhibits the PCa cells migration and invasion, and GPRC6A overexpression promotes the EMT. It is suggested that GPRC6A may serve as a potential therapeutic target for metastatic PCa.

Association between variants of IL-8 and IL-10 genes, and efficacy of transcatheter arterial chemoembolization and subsequent prognosis in patients with liver cancer.

OBJECTIVE: Our objective was to examine the association between single nucleotide polymorphisms of interleukin (IL)-8 (rs4073 and rs2227306) and IL-10 (rs1800871 and rs1800872) genes, and clinical effects of transcatheter arterial chemoembolization (TACE) and subsequent prognosis in patients with liver cancer. PATIENTS AND METHODS: 115 patients with liver cancer underwent TACE. Venous blood specimens were collected for genomic DNA extraction. The restriction fragment length polymorphism-polymerase chain reaction (RFLP-PCR) analysis was used to detect the above variants of IL-8 and IL-10 genes. In addition, blood levels of alpha fetal protein (AFP) were quantified by radioimmunoassay. Patients were followed up to uncover the association of the above genotypes with treatment efficacy and survival. RESULTS: Patients with the homozygous genotype AA or homozygous genotype TT (respectively, -251 and +781 sites) of IL-8 gene, and wild-type genotype TT or homozygous genotype AA (respectively, -819 and -592 sites) of IL-10 gene showed the best effectiveness of TACE. Furthermore, these patients also exhibited the lowest AFP levels and the longest survival after the treatment. CONCLUSIONS: Clinical efficacy of TACE and patient survival in liver cancer are associated with specific variants of IL-8 and IL-10 genes.

Seroprevalence and associated risk factors of Toxoplasma gondii infection in psychiatric patients: a case-control study in eastern China.

In recent years, the effect of Toxoplasma gondii infection on the cerebrum and neuropsychiatric patients has been increasingly highlighted. However, there is limited information about the epidemiology of T. gondii infection in psychiatric patients in Shandong province, eastern China. Therefore, through a case-control study, 445 patients hospitalized for diacrisis or treatment in Weihai, eastern China, and 445 control subjects from the general population of the same region matched by gender, age, and residence were examined with enzyme-linked immunoassays for the presence of IgG and IgM antibodies to T. gondii and associated sociodemographic and behavioural characteristics in a population of psychiatric patients. Seroprevalence of IgG antibodies to T. gondii in psychiatric patients (77/445, 17·30%) was significantly higher than in control subjects (55/445, 12·36%) (P = 0·038). Fourteen (3·15%) psychiatric patients and 10 (2·25%) control subjects had IgM antibodies to T. gondii (P = 0·408). Multivariate analysis using logic regression showed that T. gondii infection was associated with cats at home and consumption of raw/undercooked meat in psychiatric patients. Considering that most psychiatric patients usually have lower cognitive functioning and additional transmission routes related to their inappropriate behaviours that could enhance the risk of infection, psychiatric patients should be considered as a specific group of T. gondii infection.

Seroprevalence and genotype of Chlamydia in pet parrots in China.

Parrots are one of the most popular pet birds in China, and can harbour Chlamydia which has significance for human and animal health. We investigated, by indirect haemagglutination assay, the seroprevalence of Chlamydia infection in four species of parrots, namely budgerigars (Melopsittacus undulatus), lovebirds (Agapornis sp.), cockatiels (Nymphicus hollandicus) and Alexandrine parakeets (Psittacula eupatria) that were collected from Weifang and Beijing cities, North China and explored the association between potential risk factors and chlamydial seropositivity. We further determined the genotype of Chlamydia in 21 fresh faecal samples based on the ompA sequence by reconstruction of phylogenetic relationships. Of the 311 parrots examined, 35·37% (95% confidence interval 30·06-40·68) were seropositive, and species, gender, age, season and geographical location were identified as risk factors. Two PCR-positive samples represented Chlamydia psittaci genotype A. The occurrence of C. psittaci genotype A in the droppings of two pet parrots in China suggests potential environmental contamination with Chlamydiaceae and may raise a public health concern.

Seroepidemiology of Toxoplasma gondii infection in Bai and Han ethnic groups in southwestern China.

Eating raw pork and/or liver is a custom of the Bai ethnic group in China. Most people living in Dali Bai Autonomous Prefecture, Yunnan Province, southwestern China are of Bai ethnicity. Little is known of the seroprevalence of Toxoplasma gondii in Bai and Han ethnic populations in this region. In the present survey, a total of 555 and 595 blood samples were obtained from Bai and Han ethnic groups in Dali urban and rural areas, respectively. Enzyme-linked immunosorbent assay was performed to examine T. gondii IgG antibodies. Total positive rate of anti-T. gondii IgG in Bai and Han groups in this region was 21·6% (248/1150). The total seroprevalence of T. gondii was significantly higher in the Bai ethnic group (32·3%, 179/555) than in the Han ethnic group (11·6%, 69/595) (P < 0·01). The results of statistical analysis indicated that there was no significant difference between cat feeding/non-cat feeding groups in the Bai ethnic group, the most important risk factor was consumption of raw pork and/or liver for the Bai group, but feeding a cat may be the main route of T. gondii infection for the Han group. Therefore, it is essential to implement integrated strategies to prevent and control T. gondii infection in this unique region of the world.

Sequence variation and bioinformatics analysis of Toxoplasma gondii GRA16 Gene

Toxoplasmosis is caused by the intracellular protozoan Toxoplasma gondii. It is anopportunistic zoonosis in warm-blooded animals and humans, with a worldwide distribution. Toxoplasma gondii dense granule protein 16 (TgGRA16) can modulate some functions in host cells and is considered a significant virulent factor of the parasite. The present study reports sequence variation in TgGRA16 gene among T. gondii strains from different hosts and geographical locations, and the construction of phylogenetic relationships of these T. gondii strains based on sequences of TgGRA16, and analysis of B cell epitopes in TgGRA16. Our results showed that all TgGRA16 gene sequences were 1518 bp and the C+G contents ranged from 52.17% to 52.59%. Sequence variation in the TgGRA16 gene was 0-1.51%. Phylogenetic analysis revealed that TgGRA16 gene sequence could not be used to differentiate the different T. gondii genotypes. Six B cell epitopes were predicted in TgGRA16. These results indicated that TgGRA16 gene is not an ideal marker for studying genetic relationships of T. gondii isolates, but may represent a good vaccine candidate against toxoplasmosis.

Changes in cardiac aldosterone and its synthase in rats with chronic heart failure: an intervention study of long-term treatment with recombinant human brain natriuretic peptide

The physiological mechanisms involved in isoproterenol (ISO)-induced chronic heart failure (CHF) are not fully understood. In this study, we investigated local changes in cardiac aldosterone and its synthase in rats with ISO-induced CHF, and evaluated the effects of treatment with recombinant human brain natriuretic peptide (rhBNP). Sprague-Dawley rats were divided into 4 different groups. Fifty rats received subcutaneous ISO injections to induce CHF and the control group (n=10) received equal volumes of saline. After establishing the rat model, 9 CHF rats received no further treatment, rats in the low-dose group (n=8) received 22.5 μg/kg rhBNP and those in the high-dose group (n=8) received 45 μg/kg rhBNP daily for 1 month. Cardiac function was assessed by echocardiographic and hemodynamic analysis. Collagen volume fraction (CVF) was determined. Plasma and myocardial aldosterone concentrations were determined using radioimmunoassay. Myocardial aldosterone synthase (CYP11B2) was detected by quantitative real-time PCR. Cardiac function was significantly lower in the CHF group than in the control group (P<0.01), whereas CVF, plasma and myocardial aldosterone, and CYP11B2 transcription were significantly higher than in the control group (P<0.05). Low and high doses of rhBNP significantly improved hemodynamics (P<0.01) and cardiac function (P<0.05) and reduced CVF, plasma and myocardial aldosterone, and CYP11B2 transcription (P<0.05). There were no significant differences between the rhBNP dose groups (P>0.05). Elevated cardiac aldosterone and upregulation of aldosterone synthase expression were detected in rats with ISO-induced CHF. Administration of rhBNP improved hemodynamics and ventricular remodeling and reduced myocardial fibrosis, possibly by downregulating CYP11B2 transcription and reducing myocardial aldosterone synthesis.

Decreased cerebral blood flow velocity in apolipoprotein E epsilon4 allele carriers with mild cognitive impairment.

The aim of this study was to investigate the changes in cerebral blood flow velocity (CBFV) and its relation to apolipoprotein E (apoE) epsilon4 allele in mild cognitive impairment (MCI). Thirty MCI and 30 controls were assessed using Mini-Mental State Examination (MMSE) and Cambridge Cognitive Examination-Chinese version (CAMCOG-C), and then insonated in the anterior (ACA), middle (MCA) and basilar (BA) cerebral arteries using transcranial Doppler ultrasonography. Compared with controls, MCI showed significant decreases in the mean (V(m)), systolic (V(s)) and diastolic (V(d)) CBFV, bilaterally in MCA and ACA (P < 0.05-0.001), but not in BA. Compared with 17 apoE epsilon4 allele non-carriers, 13 carriers in MCI showed significant CBFV decreases, bilaterally in MCA (P < 0.05-0.001). Our findings, the decreased CBFV in apoE epsilon4 allele carriers with MCI, suggest that a large sample and longitudinal study in CBFV and cognitive changes may have the implications on early diagnosis of Alzheimer's disease.

N-NO bond dissociation energies of N-nitroso diphenylamine derivatives (or analogues) and their radical anions: implications for the effect of reductive electron transfer on N-NO bond activation and for the mechanisms of NO transfer to nitranions

The heterolytic and homolytic N-NO bond dissociation energies [i.e., deltaHhet(N-NO) and deltaHhomo(N-NO)] of 12 N-nitroso-diphenylamine derivatives (1-12) and two N-nitrosoindoles (13 and 14) in acetonitrile were determined by titration calorimetry and from a thermodynamic cycle, respectively. Comparison of these two sets of data indicates that homolysis of the N-NO bonds to generate NO* and nitrogen radical is energetically much more favorable (by 23.3-44.8 kcal/mol) than the corresponding heterolysis to generate a pair of ions, giving hints for the driving force and possible mechanism of NO-initiated chemical and biological transformations. The first (N-NO)-* bond dissociation energies [i.e., deltaH(N-NO)-* and deltaH'(N-NO)-*] of radical anions 1-*-14-* were also derived on the basis of appropriate cycles utilizing the experimentally measured deltaHhet(N-NO) and electrochemical data. Comparisons of these two quantities with those of the neutral N-NO bonds indicate a remarkable bond activation upon a possible one-electron transfer to the N-NO bonds, with an average bond-weakening effect of 48.8 +/- 0.3 kcal/mol for heterolysis and 22.3 +/- 0.3 kcal/mol for homolysis, respectively. The good to excellent linear correlations among the energetics of the related heterolytic processes [deltaHhet(N-NO), deltaH(N-NO)-*, and pKa(N-H)] and the related homolytic processes [deltaHhomo(N-NO), deltaH'(N-NO)-*, and BDE(N-H)] imply that the governing structural factors for these bond scissions are similar. Examples illustrating the use of such bond energetic data jointly with relevant redox potentials for analyzing various mechanistic possibilities for nitrosation of nitranions are presented.

The first O-NO bond energy scale in solution: heterolytic and homolytic cleavage enthalpies of O-nitrosyl carboxylate compounds

[reaction: see text] The first series of O-NO bond dissociation enthalpies was determined in solution for eight O-nitrosyl carboxylate compounds by direct titration calorimetry with a thermodynamic cycle. The derived bond energy data may serve as a quantitative guide to predict the NO binding and releasing abilities of the related amino acids.