Objective: To explore the isotemporal substitution effects among different intensities of physical activity within a 10 minute recess period on the mental health of junior high school students, aiming to provide evidence based references for targeted practical interventions. Methods: From May to November 2024, a total of 845 junior high school students from Tianjin,Taiyuan and L Liang in Shaanxi Province,Puyang in Henan Province,Xi an in Shaanxi Province,Quzhou in Zhejiang Province,and Chaoyang in Liaoning Province were selected by using a combination of stratified random sampling and convenience sampling. ActiGraph wGT3X-BT accelerometers was used to measure physical activity during a 10 minute recess period. Mental health status was assessed with the Depression Anxiety Stress Scale (DASS-21). An isotemporal substitution model was constructed in 1 minute increments to predict the effects of substituting different physical activity behaviors on students mental health. Results: During recess, sedentary behavior (SB) was predominant among junior high school students, with an average duration of [7.08(5.85,7.98)] minutes, while moderate to vigorous physical activity (MVPA) accounted for the shortest duration at [0.42(0.21,0.85)] minutes. There were statistically significant differences in MVPA,LPA and SB time between students of different genders and grades( Z/H =-9.08,-8.34,-9.51;84.87,126.82,135.27,all P <0.01). Isotemporal substitution analysis, adjusted for gender and age, showed that replacing 1 minute of SB with 1 minute of MVPA significantly improved anxiety levels ( β =-0.29, 95% CI =-0.53 to -0.04) and overall mental health ( β =-0.72, 95% CI =-1.39 to -0.04), with both results reaching statistical significance (both P <0.05). No significant effects were observed for other substitution patterns (both P >0.05). Conclusions Substituting SB with MVPA during a 10 minute recess period exerts a positive impact on the mental health of junior high school students. It is recommended to optimize the daily recess activity structure in schools to enhance students mental well being.

Mycena, a symbiont of Gastrodia elata, promotes seed germination of G. elata and plays a crucial role in the sexual reproduction of G. elata. However, the lack of genetic transformation system of Mycena blocks the research on the interaction mechanism of the two. In order to establish the protoplast transformation system of Mycena, this study analyzed the protoplast enzymatic hydrolysis system, screened the resistance markers and regeneration medium, and explored the transient transformation. After hydrolysis of Mycena hyphae with complexes enzymes for 8 h and centrifugation at 4 000 r·min~(-1), high-concentration and quality protoplast was obtained. The optimum regeneration medium for Mycena was RMV, and the optimum resistance marker was 50 mg·mL~(-1) hygromycin. The pLH-HygB-HuSHXG-GFP-HdSHXG was transformed into the protoplast of Mycena which then expressed GFP. The established protoplast transformation system of Mycena laid a foundation for analyzing the functional genes of Mycena and the molecular mechanism of the symbiosis of Mycena and G. elata.
Agaricales ; Gastrodia/genetics* ; Protoplasts ; Symbiosis/genetics* ; Transformation, Genetic

【Objective】 To explore the effects of 22 demethylases of histone H3k4, H3k9, H3k27 and H3k36 on the differentiation of fetal mice brown adipocytes, so as to provide a theoretical basis for the study of differentiation of brown adipocytes. 【Methods】 We collected the embryos from pregnant mice at stages E13.5 to E19.5, with no less than 3 fetal mice obtained at each time point. Then we isolated interscapular brown adipose tissues(BAT) for HE staining and observed the differentiation of brown adipocytes under a microscope. Realtime RT-qPCR was used to test the expression of BAT markers such as Ucp1, Cidea and Prdm16, adipocyte differentiation related gene Pparγ and 22 lysine demethylases of histone H3 in BAT. Ucp1 protein expression in BAT was detected by Western blot. 【Results】 BAT were first observed in the interscapular area of fetal mice at E14.5. Lipid droplets were observed under light microscope in the brown adipocytes of fetal mice at E18.5. Compared with those in E15.5 embryos, in E16.5 to E19.5 embryos, the expression of Ucp1, Cidea, Prdm16 and Pparγ increased(P<0.05) and Ucp1 protein expression increased(P<0.05). Of H3k4 demethylases, Jarid1a, Jarid1b, Jarid1d and Lsd1 showed decrease in expression(P< 0.05); Jarid1c expression had no statistical significance(P>0.05). Of H3k9 demethylases, Jmjd2a, Jmjd2b, Jmjd2c, Phf2, Jmjd1c and Jhdm2a showed decrease in expression(P< 0.05); Phf8, Jhdm2b and Jmjd2d showed increase in expression(P<0.05); MINA expression had no statistical difference(P>0.05). Of H3k36 demethylases, Jmjd4, Jmjd5 and Jhdm1b showed decrease in expression(P<0.05); Jhdm1a showed increase in expression(P<0.05). Of H3k27 demethylases, Jmjd3, Utx and Jhdm1d showed increase in expression(P<0.05) . 【Conclusion】 In the process of brown adipocytes differentiation, expressions of H3k4 and H3k36 demethylases were mainly downregulated and those of some H3k9 and H3k27 demethylases were upregulated. These demethylases formed a complex regulatory network with transcriptional activation and transcriptional elongation as the main effects.

An UPLC method was established for the direct determination of six major bioactive isosteroidal alkaloids, namely peimisine, imperialine, sipeimine-3-D-glucoside, verticinone, verticine and hupehenine from the bulbus of Fritillaria(Beimu), a commonly used antitussive traditional Chinese medicinal(TCM) herb. An Acquity UPLC~(TM) CSH C_(18) column(2.1 mm×100 mm, 1.7 μm) was used for all analysis. The investigated six compounds were all separated with gradient mobile phase consisting of 0.02% diethylamine-water-methanol at a flow rate of 0.3 mL·min~(-1). The temperature of sample manager was set at 20 ℃. Drift tube temperature was 45 ℃, and spray parameter was 40% with injection volume of 1 μL. Then, the further quality assessment of Beimu was carried out by cluster analysis(CA) and principal component analysis(PCA). The investigated all had good linearity(r≥0.998 9) over the tested ranges. The method is simple, accurate and reproducible, and can be used for determining the content of six major bioactive isosteroidal alkaloids.
Alkaloids/isolation & purification* ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal/chemistry* ; Fritillaria/chemistry* ; Phytochemicals/isolation & purification* ; Plant Roots/chemistry*

Inflammatory orofacial pain, in which substance P (SP) plays an important role, is closely related to the cross-talk between trigeminal ganglion (TG) neurons and satellite glial cells (SGCs). SGC activation is emerging as the key mechanism underlying inflammatory pain through different signalling mechanisms, including glial fibrillary acidic protein (GFAP) activation, phosphorylation of mitogen-activated protein kinase (MAPK) signalling pathways, and cytokine upregulation. However, in the TG, the mechanism underlying SP-mediated orofacial pain generated by SGCs is largely unknown. In this study, we investigated whether SP is involved in inflammatory orofacial pain by upregulating interleukin (IL)-1β and tumour necrosis factor (TNF)-α from SGCs, and we explored whether MAPK signalling pathways mediate the pain process. In the present study, complete Freund's adjuvant (CFA) was injected into the whisker pad of rats to induce an inflammatory model in vivo. SP was administered to SGC cultures in vitro to confirm the effect of SP. Facial expression analysis showed that pre-injection of L703,606 (an NK-1 receptor antagonist), U0126 (an inhibitor of MAPK/extracellular signal-regulated kinase [ERK] kinase [MEK] 1/2), and SB203580 (an inhibitor of P38) into the TG to induce targeted prevention of the activation of the NK-1 receptor and the phosphorylation of MAPKs significantly suppressed CFA-induced inflammatory allodynia. In addition, SP promoted SGC activation, which was proven by increased GFAP, p-MAPKs, IL-1β and TNF-α in SGCs under inflammatory conditions. Moreover, the increase in IL-1β and TNF-α was suppressed by L703, 606, U0126 and SB203580 in vivo and in vitro. These present findings suggested that SP, released from TG neurons, activated SGCs through the ERK1/2 and P38 pathways and promoted the production of IL-1β and TNF-α from SGCs, contributing to inflammatory orofacial pain associated with peripheral sensitization.

OBJECTIVE: To study the relationship between Sonic hedgehog (Shh) associated single-nucleotide polymorphism (SNP) and non-syndromic cleft lip and/or palate (NSCL/P), and to explore the risk factors of cleft lip and/or palate. Many studies suggest that the pathogenesis of NSCL/P could be related to genes that control early development, in which the Shh signaling pathway plays an important role. METHODS: Peripheral blood was collected from 197 individuals (100 patients with NSCL/P and 97 healthy controls). Haploview software was used for haplotype analysis and Tag SNP were selected, based on the population data of Han Chinese in Beijing of the international human genome haplotype mapping project. A total of 27 SNP were selected for the 4 candidate genes of SHH, PTCH1, SMO and GLI2 in the Shh signaling pathway. The genotypes of 27 SNP were detected and analyzed by Sequenom mass spectrometry. The data were analyzed by chi-squared test and an unconditional Logistic regression model. RESULTS: The selected SNP basically covered the potential functional SNP of the target genes, and its minimum allele frequency (MAF) was >0.05: GLI2 73.5%, PTCH1 91.0%, SMO 100.0%, and SHH 75.0%. It was found that the genotype frequency of SNP (rs12674259) located in SMO gene and SNP (rs2066836) located in PTCH1 gene were significantly different between the NSCL/P group and the control group. Linkage disequilibrium was also found on 3 chromosomes (chromosomes 2, 7 and 9) where the 4 candidate genes were located. However, in the analysis of linkage imbalance haplotype, there was no significant difference between the disease group and the control group. CONCLUSION In China, NSCL/P is the most common congenital disease in orofacial region. However, as it is a multigenic disease and could be affected by multiple factors, such as the external environment, the etiology of NSCL/P has not been clearly defined. This study indicates that Shh signaling pathway is involved in the occurrence of NSCL/P, and some special SNP of key genes in this pathway are related to cleft lip and/or palate, which provides a new direction for the etiology research of NSCL/P and may provide help for the early screening and risk prediction of NSCL/P.
Beijing ; Case-Control Studies ; Cleft Lip ; Cleft Palate ; Genotype ; Hedgehog Proteins ; Humans ; Nucleotides ; Polymorphism, Single Nucleotide ; Signal Transduction

As a promising new molecular imaging technique,mass spectrometry imaging(MSI) has attracted more and more attention in the field of biomedicine. A method of air flow assisted ionization-ultra high resolution mass spectrometry-based mass spectrometric imaging (AFAI-MSI) was developed to profile endogenous metabolites in rat kidney tissue in this study. Rat kidneys were collected and cut into frozen tissue sections,and then were analyzed on an AFAI-MSI system in positive ion mode using acetonitrile-isopmpyl alcohol-water (4:4:2,V/V,5 μL/min) as spray solvent,N2as spray gas(0.6 MPa) and air as assisting gas (45 L/min). The mass range and resolution were set to be 70-1000 Da and 70000, respectively. As a result,a total of 38 metabolites, including organic amines, sugars, vitamins, peptides, neurotransmitters, organic acids,phospholipids,sphingolipids,glyceride,and cholesterol esters, were identified and imaged to characterize their tissue-specific distribution in kidney tissues, and some metabolites, such as choline, acetylcoline,betaine,phoshocholine,and glycerophosphocholine were found to have distinct distribution along the cortex-medulla axis,which may be involved in the formation of osmotic pressure gradient in the kidney. The proposed ultra high resolution mass spectrometry based AFAI-MSI method could work without sample pretreatment, showed high sensitivity and wide metabolite coverage, and was expected to provide a new analytical approach for the research of in situ characterization and metabolic regulation mechanism of endogenous metabolites in kidney.

To observe the influence of different cultivation measures on the chemical constituents of Codonopsis Radix and provide reference for its reasonable cultivation, Codonopsis Radix samples cultivated by different cultivation measures were collected from the planting base in Min county,and their quality were evaluated by establishing HPLC fingerprint and determining the content of lobetyolin and Codonopsis Radix polysaccharide. The results show that different cultivation measures have an effect on the quality of Codonopsis Radix and the contents of lobetyolin and Codonopsis Radix polysaccharide are obviously different. According to the content of lobetyolin, not using Zhuanggenling>using Zhuanggenling. While, not pinching, shelving>not pinching, not shelving>pinching, shelving>pinching, not shelving. According to the content of Codonopsis Radix polysaccharide, not using Zhuanggenling>using Zhuanggenling. While, not pinching, shelving>not pinching, not shelving>pinching, not shelving>pinching, shelving. Based on the chemical quality evaluation results, the appropriate cultivation measure of Codonopsis Radix is not using Zhuanggenling, not pinching and shelving.

Objective To investigate the effects of 2-(4-di(2-chlo-roethyl) aminobutylamino)-1H-benz-[de]isoquinoline-1,3(2H)-dione( XHH) , a novel nitrogen mustard derivative, on the growth and apoptosis in human hepatoma HepG2 cells in vitro.Methods Cell pro-liferative effect was assessed by MTT assay.Changes of morphology and mitochondrial membrane potential ( MMP) were assessed by AnnexinV-FITC/Hoechst33342 , PI/Hoecsht33342 and Rh123/Hoechst33342 double staining using high content screening ( HCS ) .The expression of caspase-3 , caspase-9 , Bcl-2 , Bax was assessed by immunofluores-cence method using HCS. Results The results indicated that XHH could inhibit the proliferation of HepG2 cells, induce apoptosis, lose MMP, up-regulate the ratio of Bax/Bcl-2, increase the expression of caspase-3 , caspase-9.Conclusion XHH could inhibit proliferative of human hepatoma HepG2 cells and induce cell apoptosis via mitochon-drial pathway.

Ten compounds were isolated from the leaf of Eucommia ulmoides by means of recrystallization and chromatographic techniques such as D-101 macroporous resin, MCI resin, ODS gel, Sephadex LH-20 and Rp-HPLC. Their structures were identified by NMR spectral analyses as kaempferide 3-O-beta-D-glucoside (1), quercetin-3-O-beta-D-glucoside (2), quercetin (3), quercetin-3-O-beta-D-xylosyl-(1-->2)-beta-D-galactoside (4), kaempferol-3-O-alpha-L-rhamnosyl-(1-->6)-beta-D-glucoside (5), (2S,3S)-taxifolin 3-O-beta-D-glucoside (6) ,4-hydroxy cinnamic acid (7), (+)-cycloolivil (8), pinoresinol beta-D-glucoside (9), squalene (10). Among them compounds 1,5-7,10 were isolated from the Eucommia genus for the first time. In the DPPH free radical scavenging assay, compound 2 exhibited significant activity (IC50 13.7 micromol x L(-1)), compared with vitamin C (IC50 59.9 micromol x L(-1)); compounds 1, 3 and 9 showed moderate activity (IC50 161,137, 214 micromol x L(-1)), compared with 2,6-di-tert-butyl-4-methylphenol (IC50 236 micromol x L(-1)); compound 4 and 6 showed weak activity (IC50 264, 299 micromol x L(-1)).
Antioxidants ; chemistry ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; Eucommiaceae ; chemistry ; Magnetic Resonance Spectroscopy ; Molecular Structure ; Plant Leaves ; chemistry