Mutations in ASH1L confer susceptibility to Tourette syndrome.

Tourette syndrome (TS) is a childhood-onset neuropsychiatric disorder characterized by repetitive motor movements and vocal tics. The clinical manifestations of TS are complex and often overlap with other neuropsychiatric disorders. TS is highly heritable; however, the underlying genetic basis and molecular and neuronal mechanisms of TS remain largely unknown. We performed whole-exome sequencing of a hundred trios (probands and their parents) with detailed records of their clinical presentations and identified a risk gene, ASH1L, that was both de novo mutated and associated with TS based on a transmission disequilibrium test. As a replication, we performed follow-up targeted sequencing of ASH1L in additional 524 unrelated TS samples and replicated the association (P value = 0.001). The point mutations in ASH1L cause defects in its enzymatic activity. Therefore, we established a transgenic mouse line and performed an array of anatomical, behavioral, and functional assays to investigate ASH1L function. The Ash1l+/- mice manifested tic-like behaviors and compulsive behaviors that could be rescued by the tic-relieving drug haloperidol. We also found that Ash1l disruption leads to hyper-activation and elevated dopamine-releasing events in the dorsal striatum, all of which could explain the neural mechanisms for the behavioral abnormalities in mice. Taken together, our results provide compelling evidence that ASH1L is a TS risk gene.

Polymorphism-801G/A in the 3'-untranslated region of CXCL12 is not associated with preeclampsia in Chinese Han population.

OBJECTIVE: We investigated whether the CXCL12-801G/A polymorphism was associated with preeclampsia (PE) susceptibility in a Chinese Han population. METHODS: We examined 912 PE women and 1025 controls for the CXCL12-801G/A polymorphism by polymerase chain reaction (PCR) and correlations with clinical characteristics were examined. RESULTS: No significant differences in genotypic and allelic frequencies of CXCL12-G801A were found between cases and controls (genotype: χ2 = 2.095, p = 0.351; allele: χ2 = 1.713, p = 0.191). There were also no significant differences between early/late-onset or mild/severe PE and control groups. CONCLUSION: The results indicate that 801G/A in CXCL12 may not play a major role in pathogenesis of PE in a Chinese Han population.

[Fast measurement method based on near infrared spectroscopy in extraction process of Tianshu capsules].

Based on the near infrared spectroscopy, partial least square (PLS) method was used to respectively develop the quantitative calibration models to fast measure the contents of the total solid and ferulic acid in extraction process of Tianshu capsule extracts. The results showed that in the quantitative model of solid content, the correlation coefficients (R²) of calibration set and cross validation set were 0.967 301 and 0.947 726. The root-mean square error of calibration set (RMSEC) was 0.054 7 and root-mean square error of cross validation set (RMSECV) was 0.069 8. Besides, in the quantitative model of ferulic acid, the correlation coefficients (R²) of calibration set and cross validation set were 0.986 879 and 0.962 243. RMSEC was 1.402 6 and RMSECV was 2.400 2. When the established models were applied to on-line monitoring, the correlation coefficients of predicted results and measured values for total solid content and ferulic acid were 0.993 3 and 0.991 6; root-mean square error of predicted value (RMSEP) was 0.039 3 and 1.669 3 respectively; mean relative deviation of predicted value (RSEP) was 3.49% and 3.58%. The results indicated that the established models can be used to fast measure the contents of the total solid and ferulic acid in extraction process of Tianshu capsule extracts.

[Preliminary analysis on spectrum-efficient correlation model for anti-influenza virus of Lonicerae Japonicae Flos by partial least squares method].

Forty batches of Lonicerae Japonica Fse i collected extensively and prepared as the test solution. Their chromatographic fingerprints and anti-influenza virus IC50 value (half maximal inhibitory concentration) were determined respectively. Then Unscrambler software was used, and spectrum-efficient correlation analysis was done for chromatographic fingerprints data and IC50 data by partial least squares regression method, to establish spectrum-efficient correlation model for anti-influenza virus of Lonicerae Japonicae Flos. Then the other 10 batches of Lonicerae Japonicae Flos were used to verify the model and explore the adaptability of this spectrum-efficient correlation model based on partial least squares regression method. The mathematical model obtained R2 of 0.969489 and RM-SEC of 0.070691 for calibration set; R2 of 0.959042 and RMSECV of 0.084005 for cross validation set. The verification experiment results showed that the relative error between the predicted values and measured values was within 10% in all 10 hatches, and within 5% in 80% of them. The results showed that the established spectrum-efficient correlation model could be used to evaluate the biological activity of anti-influenza virus of Lonicerae Japonicae Flos by determining its HPLC fingerprints.

[Impurity removal technology of Tongan injection in liquid preparation process].

In order to effectively remove the invalid impurities in Tongan injection, optimize the optimal parameters of the impurity removal technology of liquid mixing process, in this paper, taking Tongan injection as the research object, with the contents of celandine alkali, and sinomenine, solids reduction efficiency, and related substances inspection as the evaluation indexes, the removal of impurities and related substances by the combined process of refrigeration, coction and activated carbon adsorption were investigated, the feasibility of the impurity removal method was definited and the process parameters were optimized. The optimized process parameters were as follows: refrigerated for 36 h, boiled for 15 min, activated carbon dosage of 0.3%, temperature 100 degrees C, adsorption time 10 min. It can effectively remove the tannin, and other impurities, thus ensure the quality and safety of products.

[Optimization of one-step pelletization technology of Jiuwei Xifeng granules by response surface methodology].

Using the qualified rates of particles as the evaluation indexes, the impact tactors of one-step pelletization technology of Jiuwei Xifeng granules were selected from six factors by the Plackett-Burman experimental design and the levels of non-significant factors were identified. According to the Plackett-Burman experimental design, choosing the qualified rates of particles and angle of repose as the evaluation indexes, three levels of the three factors were selected by Box-Behnken of central composite design to optimize the experimental. The best conditions were as follows: the fluid extract was sprayed with frequency of 29 r . min-1, inlet air temperature was 90 °C, the frequency of fan was 34 Hz. Under the response surface methodology optimized scheme, the average experimental results are similar to the predicted values, and surface methodology could be used in the optimization of one-step pelletization for Chinese materia medica.

Potential effects of soil chemical and biological properties on wood volume in Eucalyptus urophylla × Eucalyptus grandis hybrid plantations and their responses to different intensity applications of inorganic fertilizer.

Long-term and high-intensity application of inorganic fertilizer leads to a strong variation of soil characteristics. The changes in soil chemical and biological properties can significantly affect the yield of Eucalyptus plantation. However, the mechanism of soil chemical properties affecting wood volume mediated by biological factors is not clear. The purpose of this study was to identify which soil properties were affected by different fertilization intensities and to disentangle the dominant factors affecting Eucalyptus volume. After clear felling evergreen broad-leaved forest, a Eucalyptus plantation was established that was coppiced every 5 years and fertilized every year. Within this plantation, areas with different treatments were established. These treatments were a 5-year growth period (low); two times 5-year growth period (medium); and three times 5-year growth period (high). In each treatment area and in a nearby evergreen broad-leaved forest (EBLF Control), five sample plots per treatment were set up. Various biological and chemistry analyses (18 in total) were related to determining the most important path and index for optimizing Eucalyptus plantation. The analysis of variance of enzyme activity and microbial biomass showed that the soil biological characteristics decreased over 10 years of plantation, and the enzyme activity was close to the state of EBLF control in medium, while the microbial biomass failed to return to its original state during continuous planting. Redundancy analysis results show that there was a strong correlation in chemical indicators and biological characteristics. Partial least square structural equation model showed that total phosphorus, nitrate nitrogen, urease, catalase, and microbial biomass nitrogen and phosphorus were the most influential soil biochemical factors, and the indirect effect of chemical properties on volume was achieved by microorganisms through enzyme activity. Continuous planting and large-scale application of inorganic fertilizer would lead to a decrease in plantation yield and fertilizer utilization efficiency and would affect the microbial biomass and enzyme activity by destroying the stability of soil chemical properties.

The protective effect of Cu/Zn-SOD against oxidative stress after PTEN deletion.

Reactive oxygen species (ROS), antioxidant enzyme, and cancerization are related to each other. To investigate whether these factors are involved in the mechanism by which phosphatase and tensin homolog deleted on chromosome ten (PTEN) suppresses tumors. Fluorescent probes and molecular hybridizations were performed to measure ROS and antioxidant enzyme levels in cells lacking PTEN. After induction of oxidative stress by PTEN deletion, we observed an increase in the expression of the antioxidant enzyme Cu/Zn-SOD. In addition, the overall antioxidative capability of the cells was attenuated. These results suggest that negative regulation occurs through interactions among ROS, Cu/Zn-SOD, and PTEN, and such a disequilibrium between oxidation and antioxidation could contribute to the reduced antioxidative capacity in PTEN null cells. These studies will guide further investigations into the biological function of PTEN.

Study on physicochemical properties and antioxidant activity of polysaccharides from Desmodesmus armatus.

Physicochemical properties and antioxidant activities of Desmodesmus armatus polysaccharides (DAP) were studied. They were extracted by microwave-assisted constant temperature extraction and purification by DEAE-cellulose 52. Four eluents of water (DAP1), 0.25 mol/L NaCl (DAP2), 0.5 mol/L NaCl (DAP3), and 1.0 mol/L NaCl (DAP4) were collected. Four polysaccharides fractions were analyzed, and they were all composed of mannose, rhamnose, glucuronic acid, galacturonic acid, arabinose, and fucose. Gel Permeation Chromatography (GPC) analysis showed that the four polysaccharides fractions have a uniform molecular weight distribution. Scanning electron microscope showed that DAP1 had a dense structure and a smooth but uneven surface, while DAP2, DAP3, and DAP4 were amorphous solids in sheets. Oxidation in vitro experiments showed that DAP2 and DAP3 had scavenging effects on ABTS, DPPH, and hydroxyl radicals. PRACTICAL APPLICATIONS: In the determination of the antioxidant activity, it was found that the antioxidative activity of the polysaccharide of Desmodesmus armatus measured was significantly stronger than the crude polysaccharide of other microalgae. After the polysaccharide was purified, two polysaccharide fractions (DAP2 and DAP3) of Desmodesmus armatus were found to have strong scavenging ability to ABTS, DPPH, and hydroxyl radicals. They can be regarded as a new type of antioxidant, and the differences in the physicochemical properties between the parts can provide a preliminary explanation for the differences in antioxidant activity. But the connection between them needs further analysis. The Desmodesmus armatus used in the experiment is easy to cultivate and easy to obtain, which greatly increases its applicability. This research opens up new possibilities for the development of antioxidants and provides favorable evidence for the use of Desmodesmus armatus in food and feed.

[Correlation of IL-4 and IL-13 gene polymorphisms with asthma and total serum IgE levels].

OBJECTIVE: To investigate the correlation between the single nucleotide polymorphisms (SNP) of +33C/T in the promoter region of IL-4 gene and +1923C/T in intron-3 region of IL-13 gene and the susceptibility of asthma, and to study the impact of these polymorphisms upon total serum IgE levels. METHODS: The 2 polymorphisms were determined by polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP), in 150 asthmatic subjects (asthma group) and 160 healthy controls (healthy control group) of the Han nationality in Shandong province enrolled from December 2003 to June 2007. The total serum IgE levels were determined by ELISA. RESULTS: The genotype frequencies of CC, CT and TT in +33C/T sites of IL-4 gene were 43% (68/160), 35% (56/160), 22% (36/160) respectively in the controls, and 18% (27/150), 36% (54/150), 46% (69/150) respectively in the asthmatic subjects. The genotype frequencies of CC, CT and TT in +1923C/T sites of IL-13 gene were 41% (66/160), 43% (68/160), 16% (26/160) respectively in the controls, and 21% (31/150), 38% (57/150), 41% (61/150) respectively in the asthmatic subjects. The distribution of genotype in each sites between the 2 groups was significantly different (chi(2) = 27.821, 26.544 respectively, all P < 0.01). The CT and TT genotypes carried higher risks for asthma than CC genotypes (chi(2) = 21.870, 14.206 respectively, all P < 0.01). The total serum IgE levels of CC, CT and TT in +33C/T sites of IL-4 gene were (92 +/- 37), (122 +/- 45), (146 +/- 44) KU/L respectively in the controls, and (179 +/- 40), (294 +/- 51), (341 +/- 80) KU/L respectively in the asthmatic subjects. The total serum IgE levels of CC, CT and TT in +1923C/T sites of IL-13 gene were (85 +/- 31), (102 +/- 38), (144 +/- 49) KU/L respectively in the controls, and (186 +/- 65), (297 +/- 87), (363 +/- 140) KU/L respectively in the asthmatic subjects. In these 2 sites, the total serum IgE level of asthmatics was higher than that of the controls with the same genotype between the 2 groups (t = 4.653, 6.547, 7.754; and 4.673, 6.784, 8.157 respectively, all P < 0.01). The total serum IgE levels of CT, TT genotypes were higher than CC genotypes in the same group (t = 5.748, 6.253 respectively, all P < 0.01). CONCLUSIONS: There was a significant correlation between the polymorphisms of +33C/T sites of IL-4 and +1923C/T sites of IL-13 gene and susceptibility to asthma and increase of the total serum IgE. The IL-4 gene and IL-13 gene may be important candidate genes for asthma.

Visual attention modulates the asymmetric influence of each cerebral hemisphere on spatial perception.

Although the allocation of brain functions across the two cerebral hemispheres has aroused public interest over the past century, asymmetric interhemispheric cooperation under attentional modulation has been scarcely investigated. An example of interhemispheric cooperation is visual spatial perception. During this process, visual information from each hemisphere is integrated because each half of the visual field predominantly projects to the contralateral visual cortex. Both egocentric and allocentric coordinates can be employed for visual spatial representation, but they activate different areas in primate cerebral hemispheres. Recent studies have determined that egocentric representation affects the reaction time of allocentric perception; furthermore, this influence is asymmetric between the two visual hemifields. The egocentric-allocentric incompatibility effect and its asymmetry between the two hemispheres can produce this phenomenon. Using an allocentric position judgment task, we found that this incompatibility effect was reduced, and its asymmetry was eliminated on an attentional task rather than a neutral task. Visual attention might activate cortical areas that process conflicting information, such as the anterior cingulate cortex, and balance the asymmetry between the two hemispheres. Attention may enhance and balance this interhemispheric cooperation because this imbalance may also be caused by the asymmetric cooperation of each hemisphere in spatial perception.

An association study between DLGAP1 rs11081062 and EFNA5 rs26728 polymorphisms with obsessive-compulsive disorder in a Chinese Han population.

BACKGROUND: A recent genome-wide association study indicated that the single nucleotide polymorphisms (SNPs) rs11081062 in DLGAP1 and rs26728 in EFNA5 were associated with obsessive-compulsive disorder (OCD) in Caucasians. The present case-control association study assessed the global relevance of these two SNPs with respect to OCD subtypes in a Chinese Han population. METHODS: We recruited 320 OCD patients and 431 age- and sex-matched controls from a Chinese Han population. rs11081062 and rs26728 SNPs were genotyped by real-time TaqMan polymerase chain reaction, and the chi-squared test was used to compare allele and genotype frequencies of variants between the two groups. RESULTS: No significant differences were found in allele or genotype frequencies of DLGAP1 rs11081062 and EFNA5 rs26728 between the OCD and control groups. Moreover, consistently negative results were observed when classifying by sex, onset age, and comorbidity. However, on analyzing OCD subphenotypes, significant associations were observed between rs11081062 and the presence of contamination obsessions and cleaning compulsions (χ (2)=7.724, P=0.021 by genotype; χ (2)=3.745, P=0.053 by allele; and χ (2)=0.821, P=0.365 by genotype, χ (2)=27.809, P=0.000 by allele, respectively), and rs26728 with the presence of repeating compulsions (χ (2)=8.285, P=0.004 by genotype; χ (2)=7.512, P=0.006 by allele). CONCLUSION: Although we found no association between DLGAP1 rs11081062 and EFNA5 rs26728 SNPs with OCD in a Chinese Han population, obvious associations were observed with OCD subphenotypes. Therefore, it appears to be useful to divide OCD into more homogeneous subphenotypes to help understand the complex genetic basis of this disorder. Further investigations are needed to replicate these findings using larger sample sizes, different populations, and other polymorphisms.

Molecular mechanism of inhibitory effects of CD59 gene on atherosclerosis in ApoE (-/-) mice.

BACKGROUND: How to find an effective gene locus resistant to atherosclerosis has become a hotspot of today's medicine. Membrane attack complex (MAC) has proved to be related with the occurrence and development of atherosclerosis. Complement regulatory protein CD59 is a key regulator of complement MAC assembly. So this study aimed at discussing the effects of CD59 gene on occurrence and development of atherosclerosis and relative mechanism. METHODS: Apolipoprotein E knockout (ApoE (-/-)) mice were randomly divided into four groups: control group, empty plasmid-treated group, 0.5 ml CD59-treated group and 1.0 ml CD59-treated group. At the end of the 12th week, CD59 mRNA levels in whole blood were determined by RT-PCR and CD59 protein expressions were detected by western blot. The biochemical indexes in blood serum were detected. The paraffin sections of aortic root of mice were made and the degrees of atherosclerotic plaques formation were observed by hematoxylin/eosin (HE) staining. The expressions of cell apoptosis-related proteins (Bcl-2 and Fas) and plaque stability related protein (MMP-2) were detected by immunohistochemistry. Then the cell apoptosis levels were detected by TUNEL, the expression of Cyclin D1 and the mRNA level of cyclin dependent protein kinase 4 (CDK4) were detected by immunofluorescence and in situ hybridization, respectively. RESULTS: Atherosclerotic mouse model was successfully established. CD59 gene was overexpressed in blood cells and tissue cells after liposome transfection. CD59 could reduce blood lipid levels, promote the expression of anti-apoptotic Bcl-2 protein and inhibit pro-apoptotic Fas proteins, so finally lead to degradation of apoptosis levels of endothelial cells. In addition, Cyclin D1 protein and CDK4 mRNA levels were restrained by CD59 so as to inhibit the proliferation of smooth muscle cells. CD59 could inhibit the formation of atherosclerotic vulnerable plaque by suppressing the MMP-2 expression, which was further confirmed by HE staining. The anti-atherosclerotic effects were enhanced with the increase of CD59 gene dose. CONCLUSIONS: CD59 could lower blood lipid levels, positively regulate cell cycle, maintain the stability of cell proliferation and apoptosis of aorta cells, slow down the development of atherosclerotic vulnerable plaque, and finally inhibit the progress of atherosclerosis. So CD59 gene might be a new genetic locus for the therapy of atherosclerosis.

Mutation analyses of integrated HBV genome in hepatitis B patients.

Little has been learnt in the last 30 years about detection of HBV genome as well as its mutation analysis between hepatitis B fathers (HBF) and their children. In this study, we used nest polymerase chain reaction (PCR), fluorescence in situ hybridization (FISH), and DNA sequencing analysis, to examine the integrated HBV genome in paraffin-embedded testis tissues, which were taken as samples from HBF, and in peripheral blood mononuclear cells (PBMC) from 74 cases of HBFs and their children who were born after their fathers' HBV infection (caHBF). We found that HBV DNA existed in testis tissues, mainly in the basilar parts of the seminiferous tubules, and also in PBMC of HBF. It was also documented that there were point mutations of poly-loci, insertions and deletions of nucleotides in integrated HBV genomes, and the types of gene mutations in the HBFs were similar to those in caHBF. This study addresses the major types of gene mutations in integrated HBV genome in human patients and also presents reliable evidence of possible genetic transmission of hepatitis B.

[Relationship between mutation on precore region of integrated HBV DNA and p53 gene mutation in hepatocellular carcinoma].

BACKGROUND & OBJECTIVE: The chronic infection of hepatitis B virus (HBV) is a recognized risk factor for the development of hepatocellular carcinoma (HCC); however, the precise mechanism of carcinogenesis is unknown. In order to better understand the molecular mechanism of hepatocarcinogenesis, this study was designed to investigate the relationship between the gene mutation on precore region of integrated HBV DNA, the mutation of p53 gene and the development of HCC. METHODS: The integrated HBV DNA, gene mutation on precore region of HBV DNA and p53 gene mutation were detected in 80 specimens of HCC, paratumor cirrhosis, liver cirrhosis, and normal liver tissue using polymerase chain reaction (PCR) and single strand conformation polymorphism (SSCP) technique. RESULTS: (1) The positive rates of integrated HBV DNA in HCC tissues, paratumor cirrhosis tissues, cirrhotic liver tissues, and normal liver tissues were 96.43%, 96.43%, 66.67%, and 6.66%, respectively. There were significant differences of the positive rates between normal liver tissues group and each of the other three groups (P< 0.005). (2) The gene mutation rates on precore region of HBV DNA in the four groups were 70.37%, 48.15%, 25.00%, and 0%, respectively. There were significant differences of the gene mutation rates among the four groups (P< 0.05). (3) The mutation rate (57.14%)of p53 gene in HCC was significantly higher than that in liver cirrhosis (16.67%) (P< 0.05). (4) The p53 gene mutation was positively associated with positivity of integrated HBV DNA and mutation on precore region of integrated HBV DNA in different liver tissues. CONCLUSION: The HBV DNA integration and its gene mutation on precore region and p53 gene mutation probably play synergic roles in the development of HCC.