Evaluation of the inhibitory effects of drugs on the growth of Babesia gibsoni using relative quantification real-time PCR

@#To evaluate the inhibitory effects of drugs on the growth of Babesia gibsoni, relative quantification real-time PCR method was developed in this study. The 18S rRNA gene was used as a target gene for the 2–ΔΔCt method analysis. Additionally, chicken RNA was added to the parasitized blood before total RNA extraction. The chicken β-actin gene was selected as an internal control gene for the 2–ΔΔCt method analysis. The 100 µL parasitized blood samples with different percentages of parasitized erythrocytes (PPEs) (3%, 1.5%, 0.75%, 0.375% and 0.1875%) were prepared for relative quantification of B. gibsoni. Regression analysis results revealed significant linear relationships between the relative quantification value and parasitemia. 18S rRNA gene expression was significantly decreased after treatment with diminazene aceturate and artesunate in vitro drug sensitivity test. This result suggested that this relative quantification real-time PCR method can be used to evaluate the effects of drug inhibition.

Circular RNA hsa_circ_0000523 regulates the proliferation and apoptosis of colorectal cancer cells as miRNA sponge

Among the novel class of endogenous long non-coding RNAs, circular RNA (circRNA) is known as a key regulator in the development and progression of different cancers. Its function and mechanism in the tumorigenesis of colorectal cancer, however, has not been well studied. This study thus aimed to investigate potential regulation of colorectal cancer by circRNAs and the corresponding regulatory mechanism. We demonstrated that the expression of circRNA hsa_circ_0000523 (also known as circ_006229) was down-regulated in different colorectal cancer cell lines. It was also found that interference of hsa_circ_0000523 induced proliferation and suppressed apoptosis of colorectal cancer cells, the proliferation rate of which was reduced by the overexpression of hsa_circ_0000523. In addition, we found that miR-31 could recognize hsa_circ_0000523 sequence and that it acted as a "sponge" of miR-31, indirectly regulating Wnt/β-catenin signaling pathway, which was involved in the progression of colorectal cancer. The results suggested that the expression of hsa_circ_0000523 correlated to the tumorigenesis of colorectal cancer cells. In addition, as a sponge of miR-31, the low level of hsa_circ_0000523 led to activation of Wnt/β-catenin signaling pathway, inducing the subsequent progress of colorectal cancer.

Retinoic acid protects from experimental cerebral infarction by upregulating GAP-43 expression

The aim of this study was to investigate whether exogenous retinoic acid (RA) can upregulate the mRNA and protein expression of growth-associated protein 43 (GAP-43), thereby promoting brain functional recovery in a rat distal middle cerebral artery occlusion (MCAO) model of ischemia. A total of 216 male Sprague Dawley rats weighing 300–320 g were divided into 3 groups: sham-operated group, MCAO+vehicle group and MCAO+RA group. Focal cortical infarction was induced with a distal MCAO model. The expression of GAP-43 mRNA and protein in the ipsilateral perifocal region was assessed using qPCR and immunocytochemistry at 1, 3, 7, 14, 21, and 28 days after distal MCAO. In addition, an intraperitoneal injection of RA was given 12 h before MCAO and continued every day until the animal was sacrificed. Following ischemia, the expression of GAP-43 first increased considerably and then decreased. Administration of RA reduced infarction volume, promoted neurological functional recovery and upregulated expression of GAP-43. Administration of RA can ameliorate neuronal damage and promote nerve regeneration by upregulating the expression of GAP-43 in the perifocal region after distal MCAO.

A new perspective on the IIIC staging in Chinese patients with primary breast cancer: Single‑center experiences.

PURPOSE: The purpose of this retrospective study was to investigate whether Stage IIIC (TanyN3M0) breast cancer can be classified further into subgroups with different prognosis. MATERIALS AND METHODS: One hundred and thirty‑two patients with Stage IIIC breast cancer at Tianjin Medical University Cancer Institute and Hospital were analyzed. The disease‑free survival (DFS) and overall survival (OS) were calculated by Kaplan–Meier method for lymph node ratio (LNR) and the number of positive lymph node (PLN). The receiver operating characteristic curve analysis was performed to determine the optimal cut‑off value of the LNR and PLN. The univariate and multivariate analysis were applied to identify the prognostic factors. RESULTS: The results showed that the optimal cut‑off value of LNR value was 0.65, and the optimal cut‑off value of PLN was 15. The Kaplan–Meier survival analysis showed the higher value of LNR or PLN was correlated with shortened DFS (P = 0.002, P = 0.008, respectively) and OS (P < 0.001, P = 0.001, respectively). In multivariate survival analysis, the value of LNR and PLN were still remained as independent prognostic factors for DFS (P = 0.014, P = 0.013, respectively) and OS (P = 0.004, P = 0.002, respectively). CONCLUSION: These results suggest that the value of LNR or PLN could be used as a new significant prognostic biomarker for Stage IIIC breast cancer patients. Stage IIIC breast cancer patients with lower value of LNR or PLN may be down staged.

A three-phase epidemiological study of short and long sleepers in a middle-aged Chinese population: prevalence and characteristics

Epidemiological studies of short and long sleepers have not been conducted previously. We collected socioeconomic, psychological, and polysomnographic characteristics of 6501 parents (3252 men and 3249 women) of 4036 primary school children in Guangzhou city. The study data were collected in three phases. The overall prevalence of short (5 h or less) and long (10 h or more) sleep duration was 0.52 and 0.64%, respectively. Long sleepers had higher Eysenck Personality Questionnaire neuroticism scores [odds ratio (OR)=1.224, 95% confidence interval (CI)=1.047-1.409] and lower education levels (OR=0.740, 95%CI=0.631-0.849) than short sleepers. In the polysomnographic assessment, short, long, and normal sleepers (7-8 h) shared similar durations of Stage 3 sleep (short=25.7±10.7, long=20.3±7.9, and normal=28.0±12.8 min, F=1.402, P=0.181). In daytime multiple sleep latency tests, short sleepers (10/19, 52.6%) were more prone to have a short sleep latency (≤8 min) than long sleepers (2/23, 8.7%). In addition to different sleep durations, neuroticism might also contribute to differences between short and long sleepers in social achievements. Stage 3 sleep might be essential for humans. The short sleep latency (≤8 min) of short sleepers in multiple sleep latency tests should be interpreted cautiously, since it was of the same severity as required for a diagnosis of narcolepsy or idiopathic hypersomnia.

Late onset systemic lupus erythematosus with severe hypercalcaemia / Lupus eritematoso sistémico de inicio tardío con hipercalcemia severa

We report a case of a 76-year old female presenting with symptomatic severe hypercalcaemia, and subsequently diagnosed with late onset SLE due to the presence of anaemia, leucopenia, antibodies of antinuclear (ANA), anti-dsDNA, and also kidney impairment. Serum levels of FGF23 and intact-parathyroid hormone (iPTH) were low in this patient. Serum calcium, FGF23 and iPTH levels responded to steroids, which occurred simultaneously with disease activity. On follow-up, the faster increase in FGF23 than in parathyroid hormone suggested that FGF23 might be involved in the pathogenesis of hypercalcaemia in SLE.

Se reporta el caso de una mujer de 76 años de edad que se presentó con hipercalcemia sintomática severa, y a la que posteriormente le fuera diagnosticada LES de inicio tardío con presencia de anemia, leucopenia, anticuerpos antinucleares (ANA), anti-dsDNA, e insuficiencia del riñón. Los niveles séricos del factor de crecimiento fibroblástico 23 (FGF23) y la hormona paratiroidea intacta (iPTH) fueron bajos en este paciente. Los niveles de calcio séricos, FGF23 e iPTH respondieron a los esteroides, que ocurrieron simultáneamente con la actividad de la enfermedad. En el seguimiento, el hecho de que el factor FGF23 aumentara más rápidamente que la hormona paratiroidea, sugiere que el FGF23 podría estar involucrado en la patogénesis de la hipercalcemia en LES.

Human papilloma virus screening by hybrid capture II in chinese women of Jiangsu Province / Tamizaje del virus del papiloma humano mediante captura híbrida en mujeres chinas de la Provincia de Jiangsu

OBJECTIVE: To survey Human Papilloma Virus (HPV) infection in Chinese Women of Jiangsu Province and discuss the relationship between HPV and the biology of cervical cancer. METHODS: Two thousand, one hundred and fifty-three sexually active women (including 66 cases of cervical cancer) were selected for high-risk human papilloma virus DNA test with Hybrid Capture II (HCII). RESULTS: The overall HPV prevalence was 32.6% (701/2153) with higher positive rates in cervical carcinoma and Cervical Interstitial Neoplasia (CIN) [93.9% and 54.6%] respectively. For women aged 40-59 years, the overall high-risk HPV prevalence was higher than those of other age groups. Compared with CIN I, the positivity rate and viral load of HPV DNA in CIN III is much higher (80.2% vs 29.9%, 11.89 vs 0.53). Ninety-four per cent (64/66) of patients with Cervical cancer were detected to be HPV positive. There was no significant difference in HPV DNA among each clinical stage and pathologic grade. But the positive rates and the value of HPV DNA were higher in the patients with cervical interstitial incursion. Eighty per cent of patients (20/25) could become negative within six months after operation. CONCLUSIONS: High-risk HPV DNA test is effective in screening for cervical diseases. HC II is an effective method to detect HPV DNA.

OBJETIVO: Investigar la infección por el virus del papiloma humano (VPH) en las mujeres chinas de la Provincia de Jiangsu y analizar la relación entre VPH y la biología del cáncer cervical o del cuello uterino. MÉTODOS: Dos mil ciento cincuenta y tres mujeres sexualmente activas (incluyendo 66 casos de cáncer cervical) fueron seleccionadas para una prueba de ADN con el fin de detectar el virus del papiloma humano de alto riesgo mediante Captura Híbrida 2 (HC2). RESULTADOS: La prevalencia general de VPH fue 32.6% (701/2153), hallándose las tasas positivas más altas en el carcinoma cervical y la neoplasia intersticial cervical (NIC) [93.9% y 54.6%]. Para las mujeres de 40-59 años de edad, la prevalencia general de VPH de alto riesgo fue mayor que para los otros grupos etarios. En comparación con el CIN, la tasa de positividad y la carga viral de ADN del VPH en el CIN es mucho mayor (80.2% vs 29.9%, 11.89 vs 0.53). Se detectó que noventa y cuatro por ciento (64/66) de las pacientes con cáncer del cuello uterino eran VPH positivas. No hubo ninguna diferencia significativa en el ADN del VPH ADN entre cada fase clínica y el grado patológico. No obstante, tanto las tasas positivas como el valor de VPH ADN fueron más altos en las pacientes con incursión intersticial cervical. Ochenta por ciento de las pacientes (20/25) podrían volverse negativas en seis meses tras la operación. CONCLUSIONES: La prueba de ADN para la detección del virus del papiloma humano de alto riesgo es un medio efectivo para el tamizaje de las enfermedades cervicales. El HC2 es un método efectivo para detectar el ADN del VPH.

Tissue expression and subcellular localization of Mipu1, a novel myocardial ischemia-related gene

Myocardial ischemic preconditioning up-regulated protein 1 (Mipu1), a novel zinc finger protein, was originally cloned using bioinformatic analysis and 5' RACE technology of rat heart after a transient myocardial ischemia/reperfusion procedure in our laboratory. In order to investigate the functions of Mipu1, the recombinant prokaryotic expression vector pQE31-Mipu1 was constructed and transformed into Escherichia coli M15(pREP4), and Mipu1-6His fusion protein was expressed and purified. The identity of the purified protein was confirmed by mass spectrometry. The molecular mass of the Mipu1 protein was 70.03779 kDa. The fusion protein was intracutaneously injected to immunize New Zealand rabbits to produce a polyclonal antibody. The antibody titer was approximately 1:16,000. The antibody was tested by Western blotting for specificity and sensitivity. Using the antibody, it was found that Mipu1 was highly expressed in the heart and brain of rats and was localized in the nucleus of H9c2 myogenic cells. The present study lays the foundation for further study of the biological functions of Mipu1.

Carbohydrate and hydrophobic-carbohydrate recognition sites (CARS and HY-CARS) in solubilized glycosyltransferases.

Six different glycosyltransferases that are active with glycosphingolipid substrates have been purified from Golgi-membranes after solubilization with detergents. It appears that GalT-4(UDP-Gal:GlcNAc-R1 beta 1-4GalT), GalNAcT-2(UDP-Gal:Gal alpha-R2 beta 1-3GalNAcT) and FucT-2(GDP-Fuc:Gal beta GlcNAc-R3 alpha 1-2FucT) are specific for oligosaccharides bound to ceramide or to a protein moiety. These are called CARS (carbohydrate recognition sites) glycosyltransferases (GLTs). On the other hand, GalT-3(UDP-Gal:GM2 beta 1-3GalT), GalNAcT-1(UDP-GalNAc:GM3 beta 1-4GalNAcT) and FucT-3 (GDP-Fuc:LM1 alpha 1-3FucT) recognize both hydrophobic moieties (fatty acid of ceramide) as well as the oligosaccharide chains of the substrates. These GLTs are called HY-CARS (hydrophobic and carbohydrate recognition sites). D-Erythro-sphingosine (100-500 microM) modulates the in vitro activities of these GLTs. Modulation depends on the binding of D-sphingosine to a protein backbone, perhaps on more than one site and beyond transmembrane hydrophobic domains. Control of GLTs by free D-sphingosine was suggested with the concomitant discovery of ceramide glycanase in rabbit mammary tissues. The role of free sphingosine as an in vivo homotropic modulator of glycosyltransferases is becoming apparent.