[Simultaneous detection of 7 important Rickettsiales pathogens by TaqMan-probe quantitative real-time PCR].

Objective: To establish and optimize a TaqMan-probe quantitative real-time PCR (qPCR) assay for the detection of 7 important Rickettsiales pathogens and simultaneous identification of the infection types. Methods: Based on the ompB gene of Rickettsia prowazekii, Rickettsia mooseri and spotted fever group rickettsiae, the groEL gene of Orientia tsutsugamushi, the 16S rRNA of Ehrlichia chaffeensis, the gltA gene of Anaplasma phagocytophilum and the com1 gene of Coxiella burnetii, we synthesized primers and TaqMan-probes and optimized the reaction system and reaction process to same solution. The sensitivity, specificity and reproducibility of this assay were evaluated and the assay was used for the detection of simulated and actual samples. Results: The Ct value of the standard curves of the 7 pathogens showed a good linear relationship with the number of DNA copies (all R2 >0.990 0), the minimum detection limit was 10 copies/µl, showing good specificity. In the 96 tick nucleic acid extracts, Coxiella burnetii was detected in 1 sampleand spotted fever group Rickettsiae was detected in 3 samples. In the 80 blood samples from patients with undefined febrile illness, Orientia tsutsugamushi was detected in 1 sample and spotted fever group rickettsiae was detected in 2 samples. Conclusions: In this study, based on the established TaqMan-probe qPCR assay, the reaction system and reaction condition of the 7 important pathogens of Rickettsiales were optimized to the same solution. This method overcomes the shortcomings of using different reaction systems and reaction conditions for different pathogens, which can precisely identify the species of 7 important pathogens of Rickettsiales in clinical sample detections and is important for the infection type identification and laboratory detection time reduction to facilitate precise treatment of the patients.

Monitoring mild cognitive impairment of workers exposed to occupational aluminium based on quantitative susceptibility mapping.

AIM: To investigate the diagnostic value of quantitative susceptibility mapping (QSM) in mild cognitive impairment (MCI) of aluminium (Al) workers. MATERIALS AND METHODS: The basic data of 53 workers in an Al factory were collected and divided into the MCI group and normal control (NC) group by Montreal Cognitive Assessment (MoCA) scores. All participants were tested for plasma Al concentration and had magnetic resonance imaging (MRI). The QSM values of many areas of the brain were delineated and measured. Independent two-sample t-tests or non-parametric tests were used to compare the parameter values between the two groups. Spearman's correlation analysis was performed between QSM values, MoCA scores, and plasma Al concentration. The receiver operating characteristic curve and z test were performed to assess diagnostic efficacy and the best parameter. RESULTS: There was no difference in age and educational level. Plasma Al concentration of the MCI group was higher than that of NC group (p=0.057). QSM values of the left hippocampus, left dentate nucleus, right substantia nigra, and left putamen in MCI group were higher than that of NC group (p<0.05), and the left hippocampus had the best diagnostic efficacy. QSM values correlated negatively with MoCA scores. No correlation was found between QSM values and plasma Al concentration (p>0.05). CONCLUSION: QSM might be a neuroimaging marker for the diagnosis of MCI. The left hippocampus showed the best diagnostic efficacy. Plasma Al concentration of the MCI group was higher than that of the NC group. A correlation between QSM and plasma Al concentration was not found.

Long-term treadmill exercise improves spatial memory of male APPswe/PS1dE9 mice by regulation of BDNF expression and microglia activation.

Increasing evidence suggests that physical activity could delay or attenuate the symptoms of Alzheimer's disease (AD). But the underlying mechanisms are still not fully understood. To investigate the effect of long-term treadmill exercise on the spatial memory of AD mice and the possible role of ß-amyloid, brain-derived neurotrophic factor (BDNF) and microglia in the effect, male APPswe/PS1dE9 AD mice aged 4 months were subjected to treadmill exercise for 5 months with 6 sessions per week and gradually increased load. A Morris water maze was used to evaluate the spatial memory. Expression levels of ß-amyloid, BDNF and Iba-1 (a microglia marker) in brain tissue were detected by immunohistochemistry. Sedentary AD mice and wildtype C57BL/6J mice served as controls. The results showed that 5-month treadmill exercise significantly decreased the escape latencies (P < 0.01 on the 4th day) and improved the spatial memory of the AD mice in the water maze test. Meanwhile, treadmill exercise significantly increased the number of BDNF-positive cells and decreased the ratios of activated microglia in both the cerebral cortex and the hippocampus. However, treadmill exercise did not significantly alleviate the accumulation of ß-amyloid in either the cerebral cortex or the hippocampus of the AD mice (P > 0.05). The study suggested that long-term treadmill exercise could improve the spatial memory of the male APPswe/PS1dE9 AD mice. The increase in BDNF-positive cells and decrease in activated microglia might underpin the beneficial effect.

Prenatal and neonatal risk factors associated with children's developmental status at ages 4-7: lessons from the Jiangsu China birth defects prevention cohort.

BACKGROUND: There is limited information involving population data about the role of maternal health, fetal growth and neonatal health on children's developmental status at ages 4-7 years. Our aim was to determine the contribution of maternal, fetal and neonatal health to developmental status at ages 4-7 years. METHODS: In this 7-year follow-up prospective cohort study, a sample of 26,803 mothers participated in the beginning. Among their children, 19,187 voluntarily completed the development screening test or the social life ability survey, which were designed for two different age groups (<6 or ≥ 6 years old, respectively). Logistic regression analysis was used to link the data with the prenatal outcome card and interview questionnaire applied to pregnant women in previous study, to the analysis of various related factors such as demographic, socio-economic, disease and menstrual history, marriage and pregnancy care. RESULT: Univariate and multivariate logistic regression analysis were used separately in two age groups [4-6 (n = 8439) and 6-7 (n = 10,748) years old] to analyse relative factors. Maternal age of 25-30 years, maternal education of high school and greater, family income and not drinking during pregnancy were associated with higher scores in development. CONCLUSION: Both preconception and pregnancy health education and health care are the important maternal factors closely associated with children's cognitive and social competence. Public health policies for preconception care and public welfare for high-quality childcare are essential for improving children's life.

Clinicopathologic significance of S100A4 expression in osteosarcoma.

OBJECTIVE: Osteosarcoma is the most common primary malignancy, mainly arising from the metaphysis of the long bones of adolescents and young adults. Its poor prognosis is strongly associated with invasion and distant metastasis. The calcium-binding protein S100A4 promotes metastasis in several experimental animal models, including osteosarcoma (OS), and S100A4 protein expression is associated with patient outcome in a number of tumor types. In the present study, we investigated the expression of S100A4 and its clinicopathologic significance in OSs. PATIENTS AND METHODS: S100A4 were examined immunohistochemically in resected OSs from 120 patients with OS to clarify their clinicopathologic significance. Multivariate survival analyses were carried out on all investigated parameters. RESULTS: The immunohistochemical assays revealed that S1004A expression in osteosarcoma tissues was significantly higher than that in corresponding noncancerous bone tissues (p < 0.001). In addition, positive S100A4 expression more frequently occurred in osteosarcoma tissues with advanced clinical stage (p = 0.003), positive distant metastasis (p = 0.001) and poor response to chemotherapy (p = 0.04). In Kaplan-Meier analysis, only S100A4 positively stained cases showed a significantly decreased overall survival time and disease-free survival compared with negatively stained cases (both p < 0.001). On Cox multivariate analysis, positive S100A4 expression was an independent and significant prognostic factor to predict poor overall survival and disease-free survival (both p = 0.001). CONCLUSIONS: Expression of S100A4 protein in OS may be related to the prediction of metastasis potency, response to chemotherapy and poor prognosis for osteosarcoma patients, suggesting that S100A4 may serve as a prognostic marker for the optimization of clinical treatments.

The interferon-gamma (IFN-γ) +874T allele reduces the risk of hepatitis B infection in an Asian population.

Increasing evidence suggests that polymorphism of the interferon-gamma (IFN-γ) gene in the first intron at position +874 may be associated with chronic hepatitis B virus (HBV) infection and/or HBV clearance. However, the results of relevant studies have been inconsistent. To derive a more precise estimation of the association, we performed a meta-analysis. In total, 10 independent studies including 1661 chronic HBV-infected patients and 1142 controls were included in this meta-analysis. In studies following Hardy-Weinberg equilibrium (HWE), a significantly decreased risk of chronic HBV infection was associated with the IFN-γ + 874TT genotype in the overall population (TT vs AA: odds ratio (OR) = 0.714, 95% confidence interval (CI) = 0.526-0.969, P = 0.031) when compared with a spontaneously recovered population. Subgroup analysis by ethnicity revealed a similar association in Asian individuals (TT vs AA: OR = 0.706, 95% CI = 0.518-0.962, P = 0.028). Moreover, when compared with a healthy control group, the 874T allele was associated with a significant lower risk of chronic HBV infection in the overall populations (TA vs AA: OR = 0.439, 95% CI = 0.193-0.997, P = 0.049; TT + TA vs AA: OR = 0.475, 95% CI = 0.271-0.832, P = 0.009) and in Asian individuals (TA vs AA: OR = 0.862, 95% CI = 0.744-0.999, P = 0.048). In conclusion, the IFN-γ + 874TT genotype and 874T allele reduce the risk of chronic HBV infection in Asian individuals.

Relationship between IL-10 gene -1082A/G and -592C/A polymorphisms and the risk of hepatitis C infection: a meta-analysis.

Increasing evidence suggests that interleukin-10 (IL-10) gene promoter polymorphisms may be associated with chronic hepatitis C virus (HCV) infection and HCV clearance. To more precisely estimate the association between these variants and the risk of HCV infection, we performed a meta-analysis of 26 studies describing the IL-10-1082A/G, -819C/T, -592C/A genotypes, including 4039 chronic HCV infection cases and 2902 controls. When compared with a healthy population, the -1082GG allele had a 43% increased risk of chronic HCV infection in combined populations (GG vs GA + AA: odds ratio (OR) = 1.433, 95% confidence interval (CI) = 1.052-1.952, P = 0.023). In subgroup analysis by ethnicity, a significant increased risk was associated with the -1082GG genotype in the Caucasian population (GG vs AA: OR = 1.390, 95% CI: 1.108-1.744, P = 0.004; GG vs GA + AA: OR = 1.621, 95% CI: 1.267-2.075, P = 0.000). However, no significant association was found in Asian, African or Chinese populations. Moreover, a higher distribution of -592A was found in the spontaneously recovered population (AA vs CC: OR = 0.585, 95% CI = 0.387-0.884, P = 0.011; AA + AC vs CC: OR = 0.738, 95% CI = 0.551-0.988, P = 0.041; AA vs AC + CC: OR = 0.788, 95% CI = 0.664-0.935, P = 0.006) than that in the chronic HCV infection population. In conclusion, the IL-10-1082GG allele may increase the risk of chronic HCV infection in Caucasian population, and people carrying the IL-10-592A allele are more likely to clear HCV spontaneously.

Randomised control trial of tongue acupuncture versus sham acupuncture in improving functional outcome in cerebral palsy.

BACKGROUND: Traditional Chinese medicine (TCM) has been used historically in the treatment of cerebral palsy (CP). We investigated the efficacy of acupuncture in improving the motor function of children with CP. METHODS: A randomised control trial was conducted to assess the effect of tongue acupuncture (TAC) in 33 CP children. The subjects were randomised to treatment (n = 22) with TAC or control (n = 11). Clinical outcome was evaluated using the gross motor function measure (GMFM) and the pediatric evaluation of disability inventory (PEDI). RESULTS: The increase in mean GMFM score was significantly greater in the treatment than in the control group (p = 0.042). CONCLUSION: An improvement in motor function of CP subjects is seen following a short course of acupuncture.

Ruthenium complexes with a terminal hydrazido ligand. Synthesis, spectroscopy, and X-ray crystal structure.

Bis(1,1-diphenylhydrazido(1-))ruthenium(IV) porphyrins, [Ru(IV)(Por)(NHNPh2)2] (Por = TPP, TTP, 4-Cl-TPP, 4-MeO-TPP), were prepared in approximately 60% yields through the reaction of dioxoruthenium(VI) porphyrins, [Ru(VI)(Por)O2], with 1,1-diphenylhydrazine in ethanol. This new type of ruthenium complex has been characterized by 1H NMR, IR, UV-vis, and FABMS with elemental analysis. The crystal structure of [Ru(IV)(TTP)(NHNPh2)2], which reveals an eta1-coordination mode for both hydrazido axial ligands, has been determined. The average Ru-NHNPh2 distance and Ru-N-N angle were found to be 1.911(3) A and 141.1(3) degrees, respectively. The porphyrin ring exhibits a ruffling distortion that is unprecedentedly large for ruthenium complexes with simple porphyrinato ligands (such as TTP). This is probably due to the steric effect of the axial hydrazido(1-) ligands.

Reactivity of dioxoruthenium(VI) porphyrins toward amines. Synthesis and characterization of bis(arylamine)ruthenium(II), bis(arylamido)- and bis(diphenylamido)ruthenium(IV), and oxo(tert-butylimido)ruthenium(VI) porphyrins.

Reactions of dioxoruthenium(VI) porphyrins, [Ru(VI)O2(Por)], with p-chloroaniline, trimethylamine, tert-butylamine, p-nitroaniline, and diphenylamine afforded bis(amine)ruthenium(II) porphyrins, [Ru(II)(Por)(L)2] (L-p-ClC6H4NH2, Me3N, Por=TTP, 4-Cl-TPP; L=tBuNH2, Por = TPP, 3,4,5-MeO-TPP, TTP, 4-Cl-TPP, 3,5-Cl-TPP) and bis(amido)ruthenium(IV) porphyrins, [Ru(IV)(Por)(X)2] (X=p-NO2C6H4NH, Por=TTP, 4-Cl-TPP; X = Ph2N, Por = 3,4,5-MeO-TPP, 3,5-Cl-TPP), respectively. Oxidative deprotonation of [Ru(II)(Por)(NH2-p-C6H4Cl)2] in chloroform by air generated bis(arylamido)ruthenium(IV) porphyrins, [RuIV(Por)(NH-p-C6H4Cl)2] (Por=TTP. 4-Cl-TPP). Oxidation of [RuII(Por)-(NH2tBu)2] by bromine in dichloromethane in the presence of tert-butylamine and traces of water produced oxo(imido)ruthenium(VI) porphyrins, [RuVI-O(Por)(NtBu)] (Por=TPP, 3,4,5-MeO-TPP, TTP, 4-Cl-TPP, 3,5-Cl-TPP). These new classes of ruthenium complexes were characterized by 1H NMR, IR, and UV/visible spectroscopy, mass spectrometry, and elemental analysis. The structure of [Ru(IV)(TTP)(NH-p-C6H4Cl)2 . CH2Cl2 was determined by X-ray crystallography. The Ru-N bond length and the Ru-N-C angle of the Ru-NHAr moiety are 1.956(7) A and 135.8(6) degrees, respectively.

Site of macrophage inhibition of luteinizing hormone-stimulated testosterone production by purified leydig cells.

It has been observed that testicular macrophages and testicular macrophage-conditioned medium reduce LH-stimulated, but not basal, testosterone production by purified Leydig cells in vitro. In order to determine how this inhibition occurs, we have examined the effects of testicular macrophages and testicular macrophage-conditioned medium at discrete stages of the steroidogenic pathway. The lesion in steroidogenesis is located at a step beyond cAMP formation, because the addition of dibutyryl cAMP or cholera toxin did not overcome the testicular macrophage-conditioned medium inhibition of LH-stimulated steroidogenesis by Leydig cells. This effect of testicular macrophage-conditioned medium on Leydig cell testosterone production is first observed at 18 h after initiation of culture. However, subsequent additions of 22R-hydroxycholesterol, pregnenolone, dehydroepiandrosterone, or androstenedione to the Leydig cell cultures can overcome the inhibition so that, after a further 6 h of incubation, testosterone production is not significantly different from that of control Leydig cells cultured in the absence of testicular macrophage-conditioned medium. These results suggest that the block in steroidogenesis is beyond cAMP production but prior to the formation of pregnenolone, dehydroepiandrosterone, and androstenedione. Since the medium for these cultures contained lipoprotein, it is possible that the testicular macrophage-conditioned medium metabolizes the lipoprotein, making it unavailable to the Leydig cells. However, our results show that preincubation of lipoprotein with testicular macrophage-conditioned medium does not significantly alter testosterone production by the Leydig cells in the culture. It is concluded that testicular macrophage-conditioned medium affects the transport or availability of cholesterol to mitochondria prior to further steps in the steroidogenic pathway.

The effect of testicular macrophages and interleukin-1 on testosterone production by purified adult rat Leydig cells cultured under in vitro maintenance conditions.

A functional interaction between testicular macrophages and Leydig cells has been suggested. The present study attempts to clarify the interaction between purified Leydig cells and macrophages from adult male rats in coculture, employing culture conditions that maintain Leydig cell steroidogenic responsiveness in vitro. Basal Leydig cell testosterone production over 24 h was not significantly affected by coculture with macrophages, but an inhibitory effect of testicular macrophages on testosterone production by Leydig cells over 24 h was observed in the presence of increasing doses of LH from 0.125 ng/ml up to a maximally stimulating dose of 8 ng/ml. A consistent inhibitory effect was observed over a range of Leydig cell-testicular macrophage coculture ratios from 0.5:1 to 4:1 in the presence of LH (8 ng/ml). A similar inhibitory effect on maximal LH-stimulated Leydig cell testosterone production over 24 h was observed when Leydig cells were cocultured with peritoneal macrophages. Conditioned medium collected from testicular or peritoneal macrophage cultured for 24 h also inhibited LH-stimulated Leydig cell testosterone production, indicating that the effect of the macrophages was mediated by a secreted product. Inhibition of LH-stimulated testosterone production was observed also when Leydig cells were cultured in the presence of testicular macrophages for 24 h before maximal LH stimulation (8 ng/ml) for a further 24 h. Human recombinant interleukin-1 alpha and interleukin-1 beta (0.5-10 U/ml) did not significantly alter basal or LH-stimulated Leydig cell testosterone production at 24, 48, or 72 h of culture. The specific binding of 125I-human CG to Leydig cells was not affected by testicular macrophage-conditioned medium. These data demonstrate that testicular and peritoneal macrophages inhibit LH-stimulated Leydig cell testosterone production in coculture through secreted factors, acting distal to the LH receptor, and provide further support for paracrine interactions between these cell types.

CT diagnosis of lacunae.

Lacuna, a special small infarction in deep regions of the brain, is difficult to diagnose clinically. Of 182 stroke patients receiving CT scanning, 23 were diagnosed as having lacuna. 34 foci found on CT were distributed in the internal capsule, basal ganglia, thalamus and pons, with diameters varying from 3 to 22 mm. Combined CT and clinical findings showed that motor hemiplegia was present in 12 patients, pure sensory stroke in 1, sensory-motor stroke in 9, and the lock-in syndrome in 1.20 (87%) of the 23 patients partly or completely fulfilled the criteria for the lacuna syndrome. The detection of lacunae depends on the property of instrument, the method of scanning, and the thickness of slice. By showing some minor infarctions, CT has offered the possibility of diagnosis of lacunae.