Genome-wide loss of 5-hmC is a novel epigenetic feature of Huntington's disease.

5-Hydroxymethylcytosine (5-hmC) may represent a new epigenetic modification of cytosine. While the dynamics of 5-hmC during neurodevelopment have recently been reported, little is known about its genomic distribution and function(s) in neurodegenerative diseases such as Huntington's disease (HD). We here observed a marked reduction of the 5-hmC signal in YAC128 (yeast artificial chromosome transgene with 128 CAG repeats) HD mouse brain tissues when compared with age-matched wild-type (WT) mice, suggesting a deficiency of 5-hmC reconstruction in HD brains during postnatal development. Genome-wide distribution analysis of 5-hmC further confirmed the diminishment of the 5-hmC signal in striatum and cortex in YAC128 HD mice. General genomic features of 5-hmC are highly conserved, not being affected by either disease or brain regions. Intriguingly, we have identified disease-specific (YAC128 versus WT) differentially hydroxymethylated regions (DhMRs), and found that acquisition of DhmRs in gene body is a positive epigenetic regulator for gene expression. Ingenuity pathway analysis (IPA) of genotype-specific DhMR-annotated genes revealed that alternation of a number of canonical pathways involving neuronal development/differentiation (Wnt/ß-catenin/Sox pathway, axonal guidance signaling pathway) and neuronal function/survival (glutamate receptor/calcium/CREB, GABA receptor signaling, dopamine-DARPP32 feedback pathway, etc.) could be important for the onset of HD. Our results indicate that loss of the 5-hmC marker is a novel epigenetic feature in HD, and that this aberrant epigenetic regulation may impair the neurogenesis, neuronal function and survival in HD brain. Our study also opens a new avenue for HD treatment; re-establishing the native 5-hmC landscape may have the potential to slow/halt the progression of HD.

Genome-wide mapping of 5-hydroxymethylcytosine in three rice cultivars reveals its preferential localization in transcriptionally silent transposable element genes.

5-Hydroxymethylcytosine (5hmC), a modified form of cytosine that is considered the sixth nucleobase in DNA, has been detected in mammals and is believed to play an important role in gene regulation. In this study, 5hmC modification was detected in rice by employing a dot-blot assay, and its levels was further quantified in DNA from different rice tissues using liquid chromatography-multistage mass spectrometry (LC-MS/MS/MS). The results showed large intertissue variation in 5hmC levels. The genome-wide profiles of 5hmC modification in three different rice cultivars were also obtained using a sensitive chemical labelling followed by a next-generation sequencing method. Thousands of 5hmC peaks were identified, and a comparison of the distributions of 5hmC among different rice cultivars revealed the specificity and conservation of 5hmC modification. The identified 5hmC peaks were significantly enriched in heterochromatin regions, and mainly located in transposable elements (TEs), especially around retrotransposons. The correlation analysis of 5hmC and gene expression data revealed a close association between 5hmC and silent TEs. These findings provide a resource for plant DNA 5hmC epigenetic studies and expand our knowledge of 5hmC modification.

Chemical profiling of Callicarpa nudiflora and its effective compounds identification by compound-target network analysis.

Callicarpa nudiflora, belonging to the family Verbenaceae, is widely used to treat inflammation caused by bacterial infection.However, the underlying active substances of C. nudiflora against inflammation remains obscure. In this work, an ultra high-performance liquid chromatography (UHPLC) coupled with quadrupole time-of-flight mass spectrometry method was developed to characterize the ingredients in C. nudiflora, and a validated UHPLC coupled with triple quadrupole tandem mass spectrometry method was applied to quantify major components. As a result, a total of 96 chemical compounds were identified in C. nudiflora, and 26 compounds of them were further quantified in 34 batches of C. nudiflora. Based on the identified components from C. nudiflora, a compound-target network for the anti-inflammation effect was constructed by reverse docking target prediction, disease associated genes screening in DisGeNET and the protein-protein interaction from STRING. The compound-target network showed that C. nudiflora might exert anti-inflammation effect on the target of complement 3 and 5 in the pathway of cells and molecules involved in local acute inflammatory response, and 16 effective candidate compounds were found such as catalpol, acteoside, rutin, etc. This study provided an opportunity to deepen the understanding of the chemical composition and the potential anti-inflammatory mechanism of C. nudiflora.

[Application of contralateral supplementary puncture in unilateral percutaneous vertebroplasty for poor bone cement dispersion].

OBJECTIVE: To evaluate the clinical value of contralateral supplementary puncture in unilateral percutaneous vertebroplasty(PVP) with poor cement dispersion. METHODS: From January 2015 to December 2016, PVP was performed unilaterally in 285 patients(319 vertebrae) with osteoporotic compression fractures(OVCF). Contralateral percutaneous puncture was performed in 13 patients with poor cement dispersion. Among the patients, 5 cases were male and 8 patients were female, ranging in age from 63 to 88 years old; 1 case of T11, 4 cases of T12, 3 cases of L1, 2 cases of L2, 1 case of L3, and 2 cases of L5. The time from injury to operation ranged from 1 to 16 days. The VAS score, ODI score, residual low back pain and loss of vertebral height were observed. RESULTS: The VAS score and ODI score of 13 patients who underwent contralateral supplementary puncture were significantly improved (P<0.01), and there was no postoperative residual low back pain. During the follow-up period, there was no significant difference in the height loss between the two sides of the vertebral body (0.35 to 3.69 mm on the original puncture side and 0.59 to 3.66 mm on the supplementary puncture side). CONCLUSIONS: For unilateral PVP with poor cement dispersion, contralateral puncture can reduce the occurrence of postoperative residual pain; at the same time, can reduce the difference between the loss of height on both sides of the vertebral body, and then prevent the occurrence of lateral vertebral deformity, which is a safe and reliable method.

Mortality from road traffic accidents in a rapidly urbanizing Chinese city: A 20-year analysis in Shenzhen, 1994-2013.

OBJECTIVE: This study aimed to describe the trends of motorization and mortality rates from road traffic accidents and examine their associations in a rapidly urbanizing city in China, Shenzhen. METHODS: Using data from the Shenzhen Deaths Registry between 1994 and 2013, we calculated the annual mortality rates of road traffic accidents, in addition to the age- and sex-specific mortality rates and their annual percentage changes (APCs) for the period of 2000-2013. We also examined the associations between mortality rate of road traffic accidents and traffic growth with Spearman's rank correlation analysis and a log-linear model derived from Smeed's law. RESULTS: A total of 20,196 deaths due to road traffic accidents, including 14,391 (71.3%) male deaths and 5,805 (28.7%) female deaths, were recorded in Shenzhen from 1994 to 2013. The annual mortality rates in terms of deaths per population and deaths per vehicle changed in similar patterns, demonstrating an increase since 1994 and peaking in 1997, followed by a steady decrease thereafter. The decrease in mortality was faster in individuals aged 20 year or older compared to those younger than 20 years. The mortality rates in term of deaths per population were positively correlated with the total number of vehicles per kilometer of road but negatively correlated with the motorization rate in term of vehicles per population. The estimated model for deaths due to road traffic accidents in relation to the total population and the number of registered vehicles was ln (deaths/10,000 vehicles) = -1.902 × ln (vehicles/population) - 1.961. The coefficient was statistically significant (P < .001) and the coefficient of determination was 0.966, indicating a good model fit. CONCLUSIONS: We described a generally decreasing trend in the mortality rates of road traffic accidents in a rapidly urbanizing Chinese city based observations in the 20-year period from 1994 to 2013. The decreased mortality rate may be explained by the expansion of road network construction, improved road safety regulations and management, as well as more accessible ambulance services in recent years. Nevertheless, road traffic accidents remain a universal problem of great public health concern in the whole population.

Ago2 facilitates Rad51 recruitment and DNA double-strand break repair by homologous recombination.

DNA double-strand breaks (DSBs) are highly cytotoxic lesions and pose a major threat to genome stability if not properly repaired. We and others have previously shown that a class of DSB-induced small RNAs (diRNAs) is produced from sequences around DSB sites. DiRNAs are associated with Argonaute (Ago) proteins and play an important role in DSB repair, though the mechanism through which they act remains unclear. Here, we report that the role of diRNAs in DSB repair is restricted to repair by homologous recombination (HR) and that it specifically relies on the effector protein Ago2 in mammalian cells. Interestingly, we show that Ago2 forms a complex with Rad51 and that the interaction is enhanced in cells treated with ionizing radiation. We demonstrate that Rad51 accumulation at DSB sites and HR repair depend on catalytic activity and small RNA-binding capability of Ago2. In contrast, DSB resection as well as RPA and Mre11 loading is unaffected by Ago2 or Dicer depletion, suggesting that Ago2 very likely functions directly in mediating Rad51 accumulation at DSBs. Taken together, our findings suggest that guided by diRNAs, Ago2 can promote Rad51 recruitment and/or retention at DSBs to facilitate repair by HR.

N6-methyl-adenosine (m6A) in RNA: an old modification with a novel epigenetic function.

N(6)-methyl-adenosine (m(6)A) is one of the most common and abundant modifications on RNA molecules present in eukaryotes. However, the biological significance of m(6)A methylation remains largely unknown. Several independent lines of evidence suggest that the dynamic regulation of m(6)A may have a profound impact on gene expression regulation. The m(6)A modification is catalyzed by an unidentified methyltransferase complex containing at least one subunit methyltransferase like 3 (METTL3). m(6)A modification on messenger RNAs (mRNAs) mainly occurs in the exonic regions and 3'-untranslated region (3'-UTR) as revealed by high-throughput m(6)A-seq. One significant advance in m(6)A research is the recent discovery of the first two m(6)A RNA demethylases fat mass and obesity-associated (FTO) gene and ALKBH5, which catalyze m(6)A demethylation in an α-ketoglutarate (α-KG)- and Fe(2+)-dependent manner. Recent studies in model organisms demonstrate that METTL3, FTO and ALKBH5 play important roles in many biological processes, ranging from development and metabolism to fertility. Moreover, perturbation of activities of these enzymes leads to the disturbed expression of thousands of genes at the cellular level, implicating a regulatory role of m(6)A in RNA metabolism. Given the vital roles of DNA and histone methylations in epigenetic regulation of basic life processes in mammals, the dynamic and reversible chemical m(6)A modification on RNA may also serve as a novel epigenetic marker of profound biological significances.

[Influence of acupoint injection with small dose of fentanyl-droperidol mixed liquor on labor analgesia and level of stress hormone in parturient].

OBJECTIVE: To observe the clinical effect of acupoint injection with small dose of fentanyl-droperido mixed liquor at different time stages on labor analgesia, and explore its function mechanism. METHODS: One hundred and fifty cases of full-term primiparas who were intended to take vaginal delivery were randomly divided into 3 groups, 50 cases in each one. The acupoint injection with fentanyl-droperido mixed liquor at different time stages was applied in the group I, where Shenshu (BL 23) was selected in active phase and Ciliao (BL 32) was selected in the 2nd stage of labor. The subcutaneous injection with fentanyl-droperido mixed liquor was adopted in group II. The group III, which was considered as a control group, was treated with subcutaneous injection of 0.9% NaCl at the same time stage as group I and II. The blood pressure, VAS score and level of norepinephrine and adrenaline were observed at different time stages. RESULTS: Compared before the injection, the blood pressure of group III in the active phase and 2nd stage of labor was significantly increased (P < 0.05), and that in group II and III was obviously higher than that in the group I (P < 0.05). The VAS score of group I in the active phase and 2nd stage of labor was significantly lower than that in the group II and III (P < 0.01). After the injection, the level of NE and E was significantly decreased (P < 0.05), which was lower than that in the group II and III (P < 0.05). CONCLUSION: The acupoint injection with small dose of fentanyl-droperido mixed liquor at different time stages has positive effect on labor analgesia, and it could significantly relieve stress reaction of parturient during the labor. Its mechanism could be related to the reduction of stress hormone in parturient.

ALKBH4-dependent demethylation of actin regulates actomyosin dynamics.

Regulation of actomyosin dynamics by post-transcriptional modifications in cytoplasmic actin is still poorly understood. Here we demonstrate that dioxygenase ALKBH4-mediated demethylation of a monomethylated site in actin (K84me1) regulates actin-myosin interaction and actomyosin-dependent processes such as cytokinesis and cell migration. ALKBH4-deficient cells display elevated K84me1 levels. Non-muscle myosin II only interacts with unmethylated actin and its proper recruitment to and interaction with actin depend on ALKBH4. ALKBH4 co-localizes with the actomyosin-based contractile ring and midbody via association with methylated actin. ALKBH4-mediated regulation of actomyosin dynamics is completely dependent on its catalytic activity. Disorganization of cleavage furrow components and multinucleation associated with ALKBH4 deficiency can all be restored by reconstitution with wild-type but not catalytically inactive ALKBH4. Similar to actin and myosin knock-out mice, homozygous Alkbh4 mutant mice display early embryonic lethality. These findings imply that ALKBH4-dependent actin demethylation regulates actomyosin function by promoting actin-non-muscle myosin II interaction.

[Seasonal variations of soil organic carbon and microbial biomass carbon in degraded desert steppes of Inner Mongolia].

Sampling sites were installed in Damao Banner, Siziwang Banner, and Sunite Right Banner of Inner Mongolia, which represented lightly, moderately, and heavily degraded desert steppes, respectively, and surface (0-20 cm) soil samples were collected to analyze the quantitative characteristics and seasonal dynamics of soil organic carbon (SOC) and microbial biomass carbon (MBC) in these steppes. The SOC and MBC contents decreased with the increasing degradation degree of desert steppe. The total amount of soil culturable microbes in lightly and moderately degraded desert steppes was higher than that in heavily degraded desert steppe, except in summer 2006, and the MBC content and the quantity of soil culturable microbes were higher in summer and autumn than in spring and winter. Correlation analysis showed that there was a significant positive correlation between SOC and MBC (P < 0.01), suggesting that both SOC and MBC could be used as the sensitive indicators to evaluate the degradation degree of desert steppe.

[A case of human highly pathogenic avian influenza in Shenzhen, China: application of field epidemiological study].

OBJECTIVE: Based on analyzing the characteristics of a case with human avian influenza and the effects of field epidemiological study. METHODS: An emergency-response-system was started up to follow the probable human Highly Pathogenic Avian Influenza case initially detected by the "Undefined Pneumonia Surveillance System of Shenzhen". Public health professionals administered several epidemiologic investigations and giving all the contacts of the patient with a 7-day-long medical observation for temporally related influenza-like illness. Reverse transcriptase-polymerase chain reaction (RT-PCR) with primers for H5 and N1 was applied to test respiratory tract samples and/or throat swabs of the patient and all his contacts specific for the hemagglutinin gene of influenza A H5N1. Activities and strategies such as media response,notification in the public, communications with multiple related sectors, social participation and information exchange with Hong Kong were involved in field control and management. RESULTS: The patient was a male, 31 years old,with an occupation as a truck driver in a factory,and had been residing in Shenzhen for 7 years. Started with an influenza-like syndrome, the patient received treatment on the 4th day of the onset, from a clinic and on the 6th day from a regular hospital. On the 8th day of the disease course, he was confirmed by Shenzhen Center for Disease Control and Prevention as human avian flu case and was then transferred to Intensive Care Unit (ICU). On the 83rd day of commence, the patients was healed and released from the hospital. The patient had no significant exposure to sick poultry or poultry that died from the illness before the onset of the disease. The patient and five family members lived together, but no family member was affected and no contact showed positive results for H5N1. A small food market with live poultry, which was under formal supervision and before illness the patient once visited, located near his apartment. Totally, 35 swabs from live birds and bird's coops in the market for H5 nucleic acid were tested and all were negative. The influenza H5N1 virus isolated for the case was named as A/Guangdong/02/2006 (H5N1) or GD/2/06. Phylogenetic relationships and molecular characterization analysis revealed that all the segments of the H5N1 virus named GD/2/06 still belonged to avian segments. Investigation process and control measures were released to the general public through the media. Soon after the laboratory confirmation, information was released to the society, as well as Hong Kong Center for Health Protection. Local Departments of Agriculture, Industries & Business, and Entry-Exit Inspection & Quarantine Bureau together with the Public Health Department put up combined actions. A computer-based telephone survey was initiated to investigate attitudes and knowledge of residents in town, revealing that positive atmosphere dominated and no panic existed. CONCLUSION: Rapid laboratory diagnosis of the virus was the key for successful treatment and survival result of the case. Still, the pathogen was from birds resources. No human-to-human transmission was observed, however, source of infection was unclear. Field epidemiological study could offer special methods for the responses of emergency public health problems.

[Change of expression of matrix metalloproteinase-2 in the periodontal tissues of diabetes mellitus rats during orthodontic tooth movement].

OBJECTIVE: To observe the expression and distribution of matrix mentallproteinase-2 (MMP-2) in the periodontal tissues of diabetes mellitus (DM) rats during tooth movement, and to observe the affection of diabetes mellitus on the collagen metabolism. METHODS: Eighty male Sprague-Dawley rats were used. Mesial force was applied to pull the maxillary first molar. Forty rats were rendered diabetic by injection of streptozotocin. In 3 weeks after the injection, rats were subjected to lateral tooth movement. The animals were sacrificed after 0, 3, 7, 14 and 21 days respectively. Two-step immunohistochemical method was applied to localize and examine the expression of MMP-2 in periodontal tissue of rats. RESULTS: MMP-2 immunohistochemical results indicated that the MMP-2 expression increased and was observed on both sides periodontium of movement tooth, osteoclast, cementoblast, osteocyte, fibroblast and osteoblast appeared positive. According to the immunohistochemistry image analysis, experiment group changes was less obvious than control group. Dynamic changes of OD occured, reaching the minimum on the 7th day and then increasing slowly. IOD increased steadily, up to the peak on the 7th day, and then decreased, which still remained a high level on the 21st day. CONCLUSION: DM alveolar bone collagen metabolism increases. DM alveolar bone reactive potency decreases in orthodontic tooth movement, weak collagen metabolism. MMP-2's activity changes regularly, in close relation to bone remodeling, and plays an important role during the orthodontic tooth movement.

[Two sides of sublingual glands swelled following the ill fitting prosthesis used: a case report].

The patient visited for swell and ache of sublingual region. The patient's two sides of sublingual glands were swelled. The mandibular central incisors were repaired by the ill fitting prosthesis, and there was an irregular oral ulcer under it. The area of the ulcer was about 2 mm x 2 mm. The sublingual glands recovered in two days after the ill fitting prosthesis of mandibular central incisors was removed away, iodoglycerin was applied on the oral ulcer, vitamin B1, vitamin B2, prednisone and amoxicillin was taken.

[Investigation on the vectors of Borrella burgdorferi and on the identification of the isolates along China-Russia border in Eastern Heilongjiang province, China].

OBJECTIVE: To explore the fact that the east border of Heilongjiang had been a lyme disease natural focus,we investigated the species and distribution of ticks and isolated bacteria from ticks and identified genomic species of Borrelia burdorferi sensu lato. This study provided evidence for prevention and control of lyme disease. METHODS: Ticks were caught by flagging method and Direct immunofluorescence method was used to detect the rate of bacteria borne by the tick. BSK UI culture medium was used to isolate the agent and Specific McAbs were used to identify the bacteria. SDS-PAGE protein profile and PCR-RFLP method were also used to identify the species of Spirochetes. RESULTS: Ticks, collected from China-Russia border of east Heilongiiang province were classified including Ixodes persulcatus Schulze, Dermacentor sivarum Olener, Haemaphysalis concinna Kock,and Haemaphysalis japonica Kock. We found that the distributon of ticks was different under different circumstances and the predominant species were also different in different ports. The rate of bacteria borne by Iodes persulaatus Schulze was 31.4% ,by Dermacentor sivarum Olener and Haemaphysalis concinna Kock were 2.2% and 3.8%, respectively. However,it was negative for Haenaphysalis japonica Kock. Spirochetes isolated from Ixodes persulcatus Schulze were collected from Dongning and Tongjiang while Genomic species of Spirochetes, isolated from ticks of the border belonged to B. garinii. CONCLUSION: All the results showed that the east border of Heilongjiang province was the natural focus of lyme disease.