Effect of Acupuncture vs Sham Acupuncture on Patients With Poststroke Motor Aphasia: A Randomized Clinical Trial.

Importance: Motor aphasia is common among patients with stroke. Acupuncture is recommended as an alternative therapy for poststroke aphasia, but its efficacy remains uncertain. Objective: To investigate the effects of acupuncture on language function, neurological function, and quality of life in patients with poststroke motor aphasia. Design, Setting, and Participants: This multicenter, sham-controlled, randomized clinical trial was conducted in 3 tertiary hospitals in China from October 21, 2019, to November 13, 2021. Adult patients with poststroke motor aphasia were enrolled. Data analysis was performed from February to April 2023. Interventions: Eligible participants were randomly allocated (1:1) to manual acupuncture (MA) or sham acupuncture (SA) groups. Both groups underwent language training and conventional treatments. Main Outcomes and Measures: The primary outcomes were the aphasia quotient (AQ) of the Western Aphasia Battery (WAB) and scores on the Chinese Functional Communication Profile (CFCP) at 6 weeks. Secondary outcomes included WAB subitems, Boston Diagnostic Aphasia Examination, National Institutes of Health Stroke Scale, Stroke-Specific Quality of Life Scale, Stroke and Aphasia Quality of Life Scale-39, and Health Scale of Traditional Chinese Medicine scores at 6 weeks and 6 months after onset. All statistical analyses were performed according to the intention-to-treat principle. Results: Among 252 randomized patients (198 men [78.6%]; mean [SD] age, 60.7 [7.5] years), 231 were included in the modified intention-to-treat analysis (115 in the MA group and 116 in the SA group). Compared with the SA group, the MA group had significant increases in AQ (difference, 7.99 points; 95% CI, 3.42-12.55 points; P = .001) and CFCP (difference, 23.51 points; 95% CI, 11.10-35.93 points; P < .001) scores at week 6 and showed significant improvements in AQ (difference, 10.34; 95% CI, 5.75-14.93; P < .001) and CFCP (difference, 27.43; 95% CI, 14.75-40.10; P < .001) scores at the end of follow-up. Conclusions and Relevance: In this randomized clinical trial, patients with poststroke motor aphasia who received 6 weeks of MA compared with those who received SA demonstrated statistically significant improvements in language function, quality of life, and neurological impairment from week 6 of treatment to the end of follow-up at 6 months after onset. Trial Registration: Chinese Clinical Trial Registry: ChiCTR1900026740.

Comprehensive analysis of fresh tea (Camellia sinensis cv. Lingtou Dancong) leaf quality under different nitrogen fertilization regimes.

Our study investigated the impact of nitrogen fertilization at 0, 150, 300, and 450 kg/ha on the non-volatile and volatile substances, as well as gene expression in fresh leaves from Lingtou tea plants. We found that applying nitrogen at 450 kg/ha notably increased total polyphenols (TPs) and free amino acids (AAs) while decreasing the TP to AA ratio (TP/AA) and total catechins (TC) contents. Chlorophyll, caffeine (CAF) and theanine accumulated to a greater extent with nitrogen application rates of 150, 300, and 450 kg/ha, respectively, six substances - TP, CAF, TC, theanine, epigallocatechin (EGC), and AA - as key contributors to the taste quality of LTDC. Additionally, five substances with variable importance in projections (VIP) ≥ 1 and odor activation values (OAV) ≥ 1, notably linalool and cis-linalool oxide (furanoid), significantly contributed to the tea's overall aroma. Furthermore, applying 300 kg/ha nitrogen upregulated the dihydroflavonol reductase (DFR)gene, likely causing catechin decrease.

Correlations between flavor and fermentation days and changes in quality-related physiochemical characteristics of fermented Aurantii Fructus.

The effects of different fermentation times (0, 1, 2, 3, 4, and 5 days) on the physicochemical properties and flavor components of fermented Aurantii Fructus (FAF) were evaluated. Component analysis identified 66 compounds in positive ion mode and 32 compounds in negative ion mode. Flash GC e-nose results showed that propanal, (+)-limonene and n-nonanal may be the flavor characteristic components that distinguish FAF with different fermentation days. Furthermore, we found that the change of total flavonoid content was closely related to colony growth vitality. The total flavonoid content of FAF gradually decreased from 3rd day and then increased from 5th day (3rd day: 0.766 ± 0.123 mg/100 g; 4th day: 0.464 ± 0.001 mg/100 g; 5th day: 0.850 ± 0.192 mg/100 g). Finally, according to antioxidant activity correlation analysis, meranzin, (+)-limonene and total flavonoids were found to be the key substances affecting the fermentation days of FAF. Overall, the optimal fermentation time for FAF was 4 days.

Role of SIRT5 in the analgesic effectiveness of moxibustion at ST36 in mice with inflammatory pain.

Sirtuine5 (SIRT5) is an important molecule involved in the pathology of inflammatory diseases. To investigate the impact of SIRT5 on the analgesic effectiveness of moxibustion, we established a complete Freund's adjuvant- (CFA-) induced inflammatory pain in mice model. Moxibustion was applied at the Zusanli (ST36) acupoint in mice with inflammatory pain. The analgesic effectiveness was evaluated by thermal hyperalgesia and mechanical allodynia tests in the right paws after CFA injection. The expression of inflammatory cytokines, including the pro-inflammatory factors IL-1ß and TNF-α, and the anti-inflammatory factors IL-4 and TGF-ß expressions, was evaluated using by ELISA. Furthermore, SIRT5 was evaluated by immunofluorescence and western blotting. The results showed that, compared with the CFA group, both thermal and mechanical pain thresholds increased with moxibustion and the SIRT5 inhibitor MC3482 intervention at ST36. Additionally, compared to the CFA-induced group, the inflammatory mediators, including IL-1ß and TNF-α, decreased, while the anti-inflammatory cytokines IL-4 and TGF-ß increased with moxibustion and MC3482 ST36 acupoint injection. Western blot results showed a decreased expression of SIRT5 at the ST36 site with moxibustion and MC3482 injection, compared to the CFA-induced group. SIRT5 expression in the right paw of mice injected with moxibustion and MC3482 was higher than that in the CFA-induced group. This study revealed that SIRT5 expression is involved in moxibustion analgesia and may be a potential mediator in the regulation of analgesia.

Effects of sulfated polysaccharides from Laminaria japonica on regularating the gut microbiotan and alleviating intestinal inflammation in obese mice.

Due to their known health-enhancing properties, Laminaria japonica polysaccharides (LJP) may alleviate obesity via unknown mechanisms. This study aimed to investigate beneficial LJP effects and mechanism(s) of action using an animal obesity model (ICR mice fed a high-fat diet). First, LJP were confirmed to consist of sulfated polysaccharides via infrared spectroscopy. Next, LJP administration to mice was found to induce weight loss, reduce liver fat accumulation, and support healthy obesity-related blood serum indicator levels. Notably, LJP treatment significantly reduced TC and LDL levels and significantly increased HDL, LPL, UCP-2, and PPAR-α levels. Furthermore, examinations of tissues of LJP-treated mice revealed significantly reduced intestinal tissue inflammation as compared to corresponding results obtained for untreated obese controls. Additionally, LJP treatment relieved colonic shortening and reduced colonic levels of inflammatory factors TNF-α and IL-6. Further exploration of LJP treatment effects on mouse gut microbiota conducted via fecal 16S rRNA gene sequence-based gut microbiome profiling analysis revealed that LJP treatment increased the Bacteroidetes/Firmicutes ratio and increased gut abundances of probiotics Bacteroides acidifaciens, s_Lactobacillus intestinalis, and s_Lactobacillus murinus. In conclusion, these results collectively suggest that LJP use as a food supplement may alleviate obesity and related gut microbiota dysbiosis and intestinal inflammatory disorders.

Saponins from Panax japonicus alleviate adipose tissue fibrosis and metabolic dysfunction in high-fat-diet-induced obese mice.

INTRODUCTION: Adipose tissue fibrosis is a typical feature of adipose tissue dysfunction in obese individuals, which is closely related to metabolic diseases. OBJECTIVE: To explore the effect and mechanism of Saponins from Panax japonicus (SPJ) on adipose tissue fibrosis in obese mice induced by high fat diet (HFD). MATERIALS AND METHODS: We established a HFD induced obese mice model. Then the obese mice were treated with 90 mg/kg SPJ by oral gavage for 10 weeks. The levels of adipose tissue fibrosis and molecules related to signalling pathways were measured. Then the effects of SPJ on TGFß1-induced fibrosis in 3T3-L1 differentiated adipocytes were evaluated. RESULTS: SPJ reduced body weight, fat accumulation, and improved glucose and lipid metabolism in obese mice. SPJ decreased collagen deposition and expressions of fibrotic genes in epididymal white adipose tissue (eWAT) of obese mice. SPJ decreased the levels of TGFß1 protein and pSmad2, and increased the expression of PPARγ and PGC1α, thus alleviating oxidative stress in eWAT. Consistently, SPJ inhibited TGFß1-induced fibrosis in 3T3-L1 differentiated adipocytes. CONCLUSIONS: SPJ may alleviate adipose tissue fibrosis and improve obesity by inhibiting TGFß1/Smad2 and activating PPARγ/PGC1α pathway. SPJ is expected to become an efficient medicine for treatment of obesity.

Detection of weak non-covalent cation-π interactions in NGAL by single-molecule force spectroscopy.

Cation-π interaction is an electrostatic interaction between a cation and an electron-rich arene. It plays an essential role in many biological systems as a vital driving force for protein folding, stability, and receptor-ligand interaction/recognition. To date, the discovery of most cation-π interactions in proteins relies on the statistical analyses of available three-dimensional (3D) protein structures and corresponding computational calculations. However, their experimental verification and quantification remain sparse at the molecular level, mainly due to the limited methods to dynamically measure such a weak non-covalent interaction in proteins. Here, we use atomic force microscopy-based single-molecule force spectroscopy (AFM-SMFS) to measure the stability of protein neutrophil gelatinase-associated lipocalin (also known as NGAL, siderocalin, lipocalin 2) that can bind iron through the cation-π interactions between its three cationic residues and the iron-binding tri-catechols. Based on a site-specific cysteine engineering and anchoring method, we first characterized the stability and unfolding pathways of apo-NGAL. Then, the same NGAL but bound with the iron-catechol complexes through the cation-π interactions as a holo-form was characterized. AFM measurements demonstrated stronger stabilities and kinetics of the holo-NGAL from two pulling sites, F122 and F133. Here, NGAL is stretched from the designed cysteine close to the cationic residues for a maximum unfolding effect. Thus, our work demonstrates high-precision detection of the weak cation-π interaction in NGAL. Electronic Supplementary Material: Supplementary material (additional SDS-PAGE, UV-vis, protein sequences, and more experimental methods) is available in the online version of this article at 10.1007/s12274-021-4065-9.

The Physiology of Postharvest Tea (Camellia sinensis) Leaves, According to Metabolic Phenotypes and Gene Expression Analysis.

Proper postharvest storage preserves horticultural products, including tea, until they can be processed. However, few studies have focused on the physiology of ripening and senescence during postharvest storage, which affects the flavor and quality of tea. In this study, physiological and biochemical indexes of the leaves of tea cultivar 'Yinghong 9' preserved at a low temperature and high relative humidity (15-18 °C and 85-95%, PTL) were compared to those of leaves stored at ambient conditions (24 ± 2 °C and relative humidity of 65% ± 5%, UTL). Water content, chromatism, chlorophyll fluorescence, and key metabolites (caffeine, theanine, and catechins) were analyzed over a period of 24 h, and volatilized compounds were determined after 24 h. In addition, the expression of key biosynthesis genes for catechin, caffeine, theanine, and terpene were quantified. The results showed that water content, chromatism, and chlorophyll fluorescence of preserved leaves were more similar to fresh tea leaves than unpreserved tea leaves. After 24 h, the content of aroma volatiles and caffeine significantly increased, while theanine decreased in both groups. Multiple catechin monomers showed distinct changes within 24 h, and EGCG was significantly higher in preserved tea. The expression levels of CsFAS and CsTSI were consistent with the content of farnesene and theanine, respectively, but TCS1 and TCS2 expression did not correlate with caffeine content. Principal component analysis considered results from multiple indexes and suggested that the freshness of PTL was superior to that of UTL. Taken together, preservation conditions in postharvest storage caused a series of physiological and metabolic variations of tea leaves, which were different from those of unpreserved tea leaves. Comprehensive evaluation showed that the preservation conditions used in this study were effective at maintaining the freshness of tea leaves for 2-6 h. This study illustrates the metabolic changes that occur in postharvest tea leaves, which will provide a foundation for improvements to postharvest practices for tea leaves.

The Antioxidant Role of Selenium via GPx1 and GPx4 in LPS-Induced Oxidative Stress in Bovine Endometrial Cells.

This study investigated the antioxidant role of selenium (Se) in the form of selenomethionine (SLM) in LPS-induced oxidative stress via the glutathione peroxidase (GPx) enzymes and the Nrf2/HO-1 transcription factor. The impact of serum supplementation in culture media on GPxs was also studied. The bovine uterus is constantly exposed to exogenous pathogens postpartum, and the endometrium is the first contact against bacteria invasion. Endometritis is an inflammation of the endometrium and is brought about by bacterial lipopolysaccharide capable of inducing oxidative stress. The BEND cells were supplemented at the point of seeding with the following SLM concentrations 0, 100, 500, and 1000 nM for 48 h. BEND cells, cultured with or without SLM (100 nM), were initially incubated for 48 h, and then, we serum starved the SLM group for 24, 48, and 72 h. Similarly, an assay involving serum volume (0, 2, 5, and 10%) supplementation in culture media (v/v) with or without SLM (100 nM) was performed for 48 h. The BEND cells were also seeded into four experimental groups and cultured for an initial 48 h as follows: control, LPS (20 µg/mL), SLM (100 nM), and SLM + LPS groups followed by 6-h LPS treatment. The role of SLM in modulating the expressions of GPx1 and GPx4 and the Nrf2 transcription factor-related genes was assessed using qRT-PCR and Western blot techniques. The results showed serum starvation in the presence of SLM supplementation decreased the expression of GPx1 enzyme but increased GPx4 compared to the control. The addition of SLM to cell culture media in an FBS limiting condition improved the expressions of both GPx1 and GPx4. SLM supplementation promoted GPx enzymes' expressions in a serum-free media (0%) and at 2% FBS in media. However, it did not improve their expressions at 10% FBS in media than the untreated groups. Together, our data show the protective role of Se by regulating the expressions of GPx1 and GPx4 enzymes in BEND cells. It also shows that SLM promoted the expression of Nrf2 transcription factor-related genes at both the mRNA and protein levels in BEND cells during LPS stimulation.

Untargeted Metabolomics Combined with Bioassay Reveals the Change in Critical Bioactive Compounds during the Processing of Qingzhuan Tea.

Qingzhuan tea (QZT) is a typical Chinese dark tea that has a long-time manufacturing process. In the present study, liquid chromatography coupled with tandem mass spectrometry was used to study the chemical changes of tea samples during QZT processing. Untargeted metabolomics analysis revealed that the pile-fermentation and turnover (post-fermentation, FT) was the crucial stage in transforming the main compounds of QZT, whose contents of flavan-3-ols and flavonoids glycosides were decreased significantly. The bioactivities, including the antioxidant capacities and inhibitory effects on α-amylase and α-glucosidase, were also reduced after the FT process. It was suggested that although the QZT sensory properties improved following pile-fermentation and aging, the bioactivities remained restrained. Correlation analysis indicated that the main galloylated catechins and flavonoid glycosides were highly related to their antioxidant capacity and inhibitory effects on α-amylase and α-glucosidase.

Based on UPLC-Q-TOF-MS/MS, Systematic Network Pharmacology, and Molecular Docking to Explore the Potential Mechanism of Fructus Aurantii for Major Depression Disorder.

BACKGROUND: Major Depression Disorder (MDD) is a common mental disease that has become one of the world's major medical diseases. Currently, the Fructus Aurantii (FA) has been widely used to treat depression. However, the active substance ingredients and potential mechanisms of the shell antidepression have not yet been clarified. METHOD: First, we used ultraperformance liquid chromatography-quadrupole/time-of-flight tandem mass (UPLC-QTOF-MS/MS) technology to identify the chemical composition of the FA. Then, it is predicted for active ingredients, pharmaceutical disease target screening by DiscoveryStudio 2016 (DS), Metascape, and other databases, PPI network diagram, and FC core pathway. Finally, the system network pharmacology results are verified by molecular contact verification. RESULTS: Forty-six compounds in FA were identified, and twelve active ingredients were determined. Various database information, PPI network analysis of 41 intersections, and 20 core targets including DRD2, MTOR, FASP3, and PIK3P1 were integrated. Finally, the MDD treatment is indicated by molecular docking, and the most relevant potential signal pathway is the PI3K-Akt signaling pathway.

Autophagy in Gastric Mucosa: The Dual Role and Potential Therapeutic Target.

The incidence of stomach diseases is very high, which has a significant impact on human health. Damaged gastric mucosa is more vulnerable to injury, leading to bleeding and perforation, which eventually aggravates the primary disease. Therefore, the protection of gastric mucosa is crucial. However, existing drugs that protect gastric mucosa can cause nonnegligible side effects, such as hepatic inflammation, nephritis, hypoacidity, impotence, osteoporotic bone fracture, and hypergastrinemia. Autophagy, as a major intracellular lysosome-dependent degradation process, plays a key role in maintaining intracellular homeostasis and resisting environmental pressure, which may be a potential therapeutic target for protecting gastric mucosa. Recent studies have demonstrated that autophagy played a dual role when gastric mucosa exposed to biological and chemical factors. More indepth studies are needed on the protective effect of autophagy in gastric mucosa. In this review, we focus on the mechanisms and the dual role of various biological and chemical factors regulating autophagy, such as Helicobacter pylori, virus, and nonsteroidal anti-inflammatory drugs. And we summarize the pathophysiological properties and pharmacological strategies for the protection of gastric mucosa through autophagy.

Genome-wide identification of the tea plant bHLH transcription factor family and discovery of candidate regulators of trichome formation.

Leaf trichomes play vital roles in plant resistance and the quality of tea. Basic helix-loop-helix (bHLH) transcription factors (TFs) play an important role in regulating plant development and growth. In this study, a total of 134 CsbHLH proteins were identified in the Camellia sinensis var. sinensis (CSS) genome. They were divided into 17 subgroups according to the Arabidopsis thaliana classification. Phylogenetic tree analysis indicated that members of subgroups IIIc-I and IIIc-II might be associated with trichome formation. The expression patterns of CsbHLH116, CsbHLH133, CsbHLH060, CsbHLH028, CsbHLH024, CsbHLH112 and CsbHLH053 from clusters 1, 3 and 5 were similar to the trichome distribution in tea plants. CsbHLH024 and CsbHLH133 were located in the cell nucleus and possessed transcriptional activation ability. They could interact with CsTTG1, which is a regulator of tea trichome formation. This study provides useful information for further research on the function of CsbHLHs in trichome formation.

Atorvastatin reverses high cholesterol-induced cardiac remodelling and regulates mitochondrial quality-control in a cholesterol-independent manner: An experimental study.

Mitochondria are key regulators of cell fate, maintaining self-stability by a fine-tuned quality-control network including mitophagy, biogenesis, fission and fusion processes. Myocardial mitochondria can be impaired by hypercholesterolemia. Statins, such as atorvastatin, are considered the cornerstone in the management of hypercholesterolaemia primarily due to their marked cholesterol-lowering ability. The direct effect of atorvastatin on myocardial mitochondria remains unclear. We aimed to explore whether atorvastatin could attenuate myocardial mitochondrial defects induced by high cholesterol, and whether cycloastragenol, a potent telomerase activator, could be used as a potential complementary bioactive compound for obesity and hypercholesterolaemia treatment. We found that atorvastatin at a low dose (3 mg/kg) did not reduce elevated serum cholesterol, but reversed cardiac remodelling and dysfunction in C57BL/6J mice fed with high-fat diet (HFD). Atorvastatin reversed the upregulated mitophagy, mitochondrial fission and fusion, accompanied by mitochondrial biogenesis activation in HFD-fed mice hearts. Mitochondrial structural impairments were attenuated by atorvastatin in HFD-fed mice and oxidized low-density lipoprotein (ox-LDL) exposed HL-1 cardiomyocytes. The depolarized mitochondrial membrane potential and increased mitochondrial oxygen consumption rates in ox-LDL exposed HL-1 cells were recovered by atorvastatin. Furthermore, atorvastatin co-treated with cycloastragenol had better effects on reducing body weight, improving cardiac remodelling and dysfunction, and protecting mitochondria in high cholesterol. Conclusively, low-dose atorvastatin exhibited a cholesterol-independent cardioprotective effect through improving the mitochondrial quality-control network and repairing mitochondrial ultrastructure in high cholesterol. Atorvastatin plus cycloastragenol supplement therapy has a better effect on treating obesity and hypercholesterolaemia.

OsPPR939, a nad5 splicing factor, is essential for plant growth and pollen development in rice.

KEY MESSAGE: P-subfamily PPR protein OsPPR939, which can be phosphorylated by OsS6K1, regulates plant growth and pollen development by involving in the splicing of mitochondrial nad5 introns 1, 2, and 3. In land plants, pentatricopeptide repeat (PPR) proteins play key roles in mitochondrial group II intron splicing, but how these nucleus-encoded proteins are imported into mitochondria is unknown. To date, a few PPR proteins have been characterized in rice (Oryza sativa). Here, we demonstrate that the mitochondrion-localized P-subfamily PPR protein OsPPR939 is required for the splicing of nad5 introns 1, 2, and 3 in rice. Complete knockout or partial disruption of OsPPR939 function resulted in different degrees of growth retardation and pollen sterility. The dramatically reduced splicing efficiency of these introns in osppr939-4 and osppr939-5 led to reduced mitochondrial complex I abundance and activity and enhanced expression of alternative respiratory pathway genes. Complementation with OsPPR939 rescued the defective plant morphology of osppr939-4 and restored its decreased splicing efficiency of nad5 introns 1, 2, and 3. Therefore, OsPPR939 plays crucial roles in plant growth and pollen development by splicing mitochondrial nad5 introns 1, 2, and 3. More importantly, the 12th amino acid Ser in the N-terminal targeting sequence of OsPPR939 is phosphorylated by OsS6K1, and truncated OsPPR939 with a non-phosphorylatable S12A mutation in its presequence could not be imported into mitochondria, suggesting that phosphorylation of this amino acid plays an important role in the mitochondrial import of OsPPR939. To our knowledge, the 12th residue Ser on OsPPR939 is the first experimentally proven phosphorylation site in PPR proteins. Our results provide a basis for investigating the regulatory mechanism of PPR proteins at the post-translational level.

Research strategies for endophytes in medicinal plants based on high-throughput sequencing and traditional culture and isolation methods / 中国中药杂志

The research on endophytes of medicinal plants mainly relies on the traditional culture and isolation methods. Because of their functions such as promoting host growth, improving stress resistance, promoting the accumulation of medicinal active ingredients or directly producing medicinal active ingredients, the endophytes of medicinal plants have gradually attracted wide attention. However, it was found that the strains isolated by traditional methods were not the true dominant endophytes of medicinal plants by comparing the results of traditional culture isolation with high-throughput sequencing. The blind and random nature of traditional methods leads to the lack of standards in terms of medium selection, culture time and interaction between species. On the contrary, high-throughput sequencing technology is an emerging molecular biology technology developed in recent decades. Due to its high resolution level and indepen-dent culture, it can be used for thorough analysis of the community structure and diversity of environmental microorganisms. Therefore, we proposed the strategy of using high-throughput sequencing technology to guide the traditional culture and isolation of endophytes from medicinal plants. Firstly, the endophytic structure and diversity of medicinal plants were completely clear by high-throughput sequencing technology, and the dominant endophytes of the host were unequivocal. Then according to the characteristics of each dominant endophytes design or query suitable medium for its growth to culture and isolation. Finally, the function of the isolates was studied. This method can prevent researchers from missing out on the important functional strains of the host, expand the research scope of endophytes of medicinal plants, and facilitate the in-depth excavation and utilization of endophytes of medicinal plants.

A potential mechanism associated with lead-induced spermatogonia and Leydig cell toxicity and mitigative effect of selenium in chicken.

Lead (Pb) is a toxic heavy metal pollutants and can damage male reproductive function. Selenium (Se) possesses an ability of antagonizing Pb toxicity. However, biological events in the process of Pb toxicity and mitigative effect of Se are not well understood. The aim of present research was to investigate potential mechanism of Se against Pb toxicity from the perspective of oxidative stress, heat shock response and autophagy in the spermatogonia and Leydig cell of chicken. The cells from one-day-old male Hyline chickens were treated with Se (0.5 µmol/L) and/or Pb (20 µmol/L) for 24 h, respectively. Cell viability, cell ultrastucture, Pb and Se concentrations, testosterone level, oxidative stress indicators and relative expression of heat shock proteins (HSPs) and autophagy-related genes were measured. The results showed that spermatogonia was more tolerant to Pb than Leydig cell; cell injury was confirmed via histological assessment, cell viability and testosterone level; oxidative stress was further indicated by the decrease of catalase, glutathione peroxidase, glutathione-s-transferase and superoxide dismutase activities and the increase of malondialdehyde and reactive oxygen species contents. Pb increased expression of HSPs (27, 40, 60, 70 and 90). Meanwhile Pb induced autophagy through up-regulation of autophagy-related proteins 5, Beclin 1, Dynein, light chain 3 (LC3)-I and LC3-II and down-regulation of mammalian target of rapamycin in two type cells of chicken. However, Se intervention mitigated the aforementioned alterations caused by Pb. In conclusion, Pb led to oxidative stress, which triggered heat shock response and autophagy; Se administration mitigated reproductive toxicity of Pb through strengthening antioxidant defense in the spermatogonia and Leydig cell of chicken.

[Moxa nitrogen content determination and its application in moxa floss grade identification].

Artemisia argyi samples were collected from different areas in Hubei Qichun and medicinal botanical garden of Hubei University of Traditional Chinese Medicine, and samples were purchased from different areas in Qichun medicinal market. The same method was used to prepare A. argyi samples from different areas into moxa floss of different leaf velvet ratio. The nitrogen content of A. argyi was determined by Kjeldahl method. The results show that there was an inverse relationship between the content of nitrogen and the level of the leaf velvet ratio grades of moxa floss and nitrogen content. Therefore, six functional models were established to illustrate the relationship between the grades of leaf velvet ratio and nitrogen content. The accuracy of the functional models was evaluated by ■. The cubic function model and exponential function model are selected as the best models and the formula is: Y_(grades)=-4.26 N~3+26.09 N~2-61.09 N+55.84, Y_(grades)=63.68×10~(-1.374N), with R~2 values of 0.976 5 and 0.969 6, respectively. The accuracy reached above 95% of the two function models when verified by laboratory-made moxa floss, while the accuracy of the models was 85.44% when verifed by commercial moxa floss. These results indicate that the model could be applied to the identification of the grades of moxa floss.

Effect of Electroacupuncture at GV20 on Sleep Deprivation-Induced Depression-Like Behavior in Mice.

Accumulating evidence suggests that sleep deprivation (S-Dep) is a critical risk factor for depression. Electroacupuncture (EA) treatment has been reported to ameliorate posttraumatic stress disorder- (PSTD-) like behavior and enhance hippocampal neurogenesis. However, whether EA treatment has any beneficial effect on S-Dep-induced depression-like behavior is still unknown. In the present study, we focused on whether EA at Baihui (GV20) can ameliorate the deterioration effect of S-Dep in mice. Mice were randomly divided into normal, S-Dep, S-Dep + EA, and S-Dep + sham EA groups. Cognitive behavior test and in vitro assay were performed separately to avoid the influence of behavior test on synaptic transmission and protein expression. Depression-like behaviors were determined by forced swimming test (FST), tail suspension test (TST), and Morris water maze (MWM). Neurogenesis was identified by BrdU, DCX, and NeuN immunofluorescence staining. In vitro long-term potentiation was detected by high frequency stimulation (HFS) at Schaffer collateral-CA1 synapses in hippocampal slices. Brain-derived neurotropic factor (BDNF) and tropomyosin receptor kinase B (TrkB) protein expression level were assayed by western blot. Our results indicated that D-Sep mice demonstrated depression-like behaviors determined by prolonged immobility duration in FST and TST; D-Sep mice also manifested spatial memory retention deficit in MWM. Furthermore, EA treatment ameliorated D-Sep-induced depression-like behaviors and spatial memory retention deficit. Mechanically, EA treatment alleviated neuron progenitor cell proliferation and differentiation, ameliorated the field excitatory postsynaptic potentials (fEPSPs) slope impaired by S-Dep, and elevated BDNF/TrkB protein expression. Taken together, our data suggested that EA treatment has a protective effect on S-Dep-induced depression-like behavior and cognitive impairment, which may be through regulating BDNF/TrkB protein expression.

Anti-inflammatory effect of selenium on lead-induced testicular inflammation by inhibiting NLRP3 inflammasome activation in chickens.

Lead (Pb) is a deleterious environmental pollutant that is toxic to testes. Selenium (Se) possesses antioxidant and anti-inflammatory properties. Nucleotide-binding domain, leucine-rich-containing family, pyrin-domain containing-3 (NLRP3) inflammasome is involved in inflammatory response. However, the function of NLRP3 inflammasome in antagonistic effect of Se on inflammation caused by Pb remains unknown. The purpose of this research is to identify anti-inflammatory role of Se on testicular toxicity induced by Pb with an emphasis on oxidative stress, inflammation and NLRP3 signaling pathway in chicken. In present study, sixty seven-day-old Hyline male chickens were assigned into four groups. The feeding program consisted of a commercial diet (0.49 mg/kg Se), a Se-supplemented diet (1 mg/kg Se), a Pb-supplemented diet (0.49 mg/kg Se and 350 mg/kg Pb) and a Se-supplemented and Pb-supplemented diet (1 mg/kg Se and 350 mg/kg Pb), respectively. On the 12th week, blood was collected to measure serum testosterone level and testicular tissues were removed to determine Se and Pb concentrations, testicular function, histological structure, oxidative stress indicators and inflammation-related factors (Nuclear factor-kappaB, tumor necrosis factor-α, cyclooxygenase-2, NLRP3, apoptosis-associated speck-like protein containing a caspase recruitment domain, caspase-1, interluekin (IL)-1ß, IL-6, IL-18 and interferon-γ). The experimental results showed that after Pb administration, testicular injury was confirmed via histological assessment; testicular dysfunction were further indicated by decreased testosterone level and mRNA expression of steroidogenic acute regulatory protein, cytochrome P450 side-chain cleavage, 3ß-hydroxysteroid dehydrogenase and 17ß-hydroxysteroid dehydrogenase. Moreover, NLRP3 signaling pathway activated by Pb-caused oxidative stress was up-regulated accompanied by promotion in reactive oxygen species, nitric oxide, inducible nitric oxide synthase and malondialdehyde and reduction in antioxidants including glutathione peroxidase and glutathione s-transferase. Se administration ameliorated testicular tissue injury, testicular function, oxidative stress and inflammation. In conclusion, Se exhibited antagonistic role in Pb-induced testicular injury via enhancing antioxidant system and inhibiting inflammation in chickens.