A stabilization rule for metal carbido cluster bearing µ3-carbido single-atom-ligand encapsulated in carbon cage.

Metal carbido complexes bearing single-carbon-atom ligand such as nitrogenase provide ideal models of adsorbed carbon atoms in heterogeneous catalysis. Trimetallic µ3-carbido clusterfullerenes found recently represent the simplest metal carbido complexes with the ligands being only carbon atoms, but only few are crystallographically characterized, and its formation prerequisite is unclear. Herein, we synthesize and isolate three vanadium-based µ3-CCFs featuring V = C double bonds and high valence state of V (+4), including VSc2C@Ih(7)-C80, VSc2C@D5h(6)-C80 and VSc2C@D3h(5)-C78. Based on a systematic theoretical study of all reported µ3-carbido clusterfullerenes, we further propose a supplemental Octet Rule, i.e., an eight-electron configuration of the µ3-carbido ligand is needed for stabilization of metal carbido clusters within µ3-carbido clusterfullerenes. Distinct from the classic Effective Atomic Number rule based on valence electron count of metal proposed in the 1920s, this rule counts the valence electrons of the single-carbon-atom ligand, and offers a general rule governing the stabilities of µ3-carbido clusterfullerenes.

The Long-Term Effect of Maternal Iron Levels in the Second Trimester on Mild Thinness among Preschoolers: The Modifying Effect of Small for Gestational Age.

The supplementation of multiple micronutrients throughout pregnancy can reduce the risk of adverse birth outcomes and various diseases in children. However, the long-term effect of maternal multiple micronutrient levels in the second trimester on the overall development of preschoolers remains unknown. Therefore, 1017 singleton mother-infant pairs and 6-year-old preschoolers were recruited based on the China-Wuxi Birth Cohort Study. Meanwhile, information on the demographic characteristics of pregnant women and preschoolers, maternal copper, calcium, iron, magnesium, and zinc levels in whole blood during the second trimester, and neonatal outcomes, were collected. We aimed to investigate the long-term impact of maternal copper, calcium, iron, magnesium, and zinc levels in the second trimester on mild thinness among 6-year-old preschoolers, and the modifying effect of small for gestational age (SGA), within the Chinese population. Multiple logistic regression models revealed that high-level maternal iron in the second trimester reduced the risk of mild thinness [adjusted OR: 0.46 (95% CI: 0.26, 0.80)] among 6-year-old preschoolers. However, no significant association was found for the remaining four maternal essential metal elements. Additionally, the restricted cubic spline function showed that the risk of mild thinness decreased when maternal iron concentration exceeded 7.47 mmol/L in whole blood during the second trimester. Furthermore, subgroup analysis indicated that the long-term protective effect of high-level maternal iron on mild thinness was only observed in SGA infants. Summarily, high-level maternal iron in the second trimester distinctly lowers the risk of mild thinness among 6-year-old preschoolers, especially in preschoolers with birth outcomes of SGA. Our findings offer evidence for the implementation of iron supplementation in the second trimester as a preventive measure against mild thinness in children.

[Status of Heavy Metal in Organic Fertilizers in Main Tea Growing Regions of China].

To provide guidance for the safe use of organic fertilizers and improve soil quality and tea safety, it is necessary to conduct systematic analyses of the heavy metal content of organic fertilizers applied in the main tea producing areas of China. In this study, we analyzed the heavy metal contents in organic fertilizer samples collected from 2017 to 2019. The risks of collected organic fertilizers from different areas and sources were calculated. The results showed that the average concentrations of ω(As), ω(Hg), ω(Pb), ω(Cd), ω(Cr), ω(Cu), ω(Zn), and ω(Ni) in the collected organic fertilizers were 4.60, 0.22, 27.1, 0.78, 27.9, 58.3, 250.1, and 16.3 mg·kg-1, respectively. According to the assessment standard in NY/T 525- 2021, the over-limit rates of As, Hg, Pb, Cd, and Cr were 6.19%, 1.33%, 4.42%, 4.42%, and 1.33%, respectively. With respect to the area, the qualified rates were 100% in Shaanxi, Jiangsu, Anhui, Fujian, and Guangxi; 80%-90% in Shandong, Zhejiang, Hubei, Sichuan, Yunnan, and Guangdong; and only 54.5% in Jiangxi. The qualified rates of sources were 100% in rapeseed cake, soybean cake, and pig manure; 95.8% in sheep manure; 91.7% in cow manure; 90.7% in chicken manure; 87.2% in manure of other animals; 82.4% in the mixture of plant and animal sources; 65.2% in other plant sources; and 63.6% in other sources. According to the recommended application rate, the accumulation rate of heavy metals in soil with pig manure, cow manure, chicken manure, and sheep manure would be much higher than that with rapeseed cake and soybean cake. The average accumulation rate of organic fertilizer from animal sources was 7-30 times higher than that from plant sources. Therefore, it is recommended to use rapeseed cake or soybean cake fertilizer in tea plantation and to increase the supervision of heavy metal accumulation in soil and tea in those high-risk areas.

[Tea Polyphenols Regulate Renin-angiotensin-aldosterone System and Transforming Growth Factor-ß1/Smads Signaling Pathway in Heart Failure Rats].

Objective To investigate the effects of tea polyphenols on the renin-angiotensin-aldosterone system and the transforming growth factor-ß1(TGF-ß1)/Smads signaling pathway in heart failure rats.Methods SD rats were randomly assigned into a sham group,a model group,a captopril group(PC group),and a tea polyphenol group(TP group).The left anterior descending coronary artery was ligated with silk thread to establish the rat model of heart failure after myocardial infarction in the model group,PC group,and TP group,while it was not ligated in the sham group.Echocardiography was used to detect cardiac function.HE staining and Masson staining were conducted for the observation of myocardial pathological changes and myocardial fibrosis,respectively.Immunohistochemistry was employed to detect the expression of collagen Ⅰ and collagen Ⅲ.ELISA kits were used to measure the levels of angiotensin Ⅱ(AngⅡ),aldosterone(ALD),plasma renin activity(PRA),interleukin-1ß(IL-1ß),IL-6,and tumor necrosis factor-α(TNF-α).Western blotting was employed to determine the protein levels of TGF-ß1,phosphorylated Smad2(p-Smad2),Smad2,p-Smad3,and Smad3.Results Compared with the sham group,the model group showed disordered myocardial cells with obvious inflammatory cell infiltration,increased degree of myocardial fibrosis(t=9.748,P=0.001),elevated levels of collagen Ⅰ(t=11.754,P=0.001) and collagen Ⅲ(t=10.573,P=0.001),decreased ejection fraction(EF)(t=13.174,P=0.002) and left ventricular short axis shortening rate(LVFS)(t=11.853,P=0.001),and up-regulated expression of AngⅡ(t=4.246,P=0.001),ALD(t=5.385,P=0.004),PRA(t=4.386,P=0.004),IL-1ß(t=4.393,P=0.001),IL-6(t=6.375,P=0.002),and TNF-α(t=4.753,P=0.002),and up-regulated protein levels of TGF-ß1(t=6.365,P=0.001),p-Smad2/Smad2(t=13.755,P=0.001),and p-Smad3/Smad3(t=11.657,P=0.002).Compared with the model group,PC and TP alleviated the myocardial pathological changes,decreased the left ventricular end-diastolic diameter(LVEDd)(t=6.367,P=0.003 and t=5.264,P=0.003),left ventricular end-systolic diameter(LVEDs)(t=5.253,P=0.002 and t=5.974,P=0.001),heart mass index(HMI)(t=5.012,P=0.007 and t=4.953,P=0.005),left ventricular mass index(LVMI)(t=5.531,P=0.003 and t=5.483,P=0.004),and the degree of myocardial fibrosis(t=6.734,P=0.001 and t=5.362,P=0.001).Furthermore,they lowered the levels of collagen Ⅰ (t=5.373,P=0.001 and t=4.364,P=0.001) and collagen Ⅲ(t=6.764,P=0.001 and t=4.579,P=0.001),increased EF(t=11.264,P=0.002 and t=10.356,P=0.001) and LVFS(t=8.246,P=0.002 and t=7.824,P=0.001),and down-regulated the expression of AngⅡ(t=3.126,P=0.001 and t=2.853,P=0.001),ALD(t=3.854,P=0.004 and t=3.164,P=0.004),PRA(t=3.126,P=0.004 and t=3.063,P=0.004),IL-1ß(t=2.964,P=0.001 and t=2.765,P=0.001),IL-6(t=4.865,P=0.002 and t=4.275,P=0.002),and TNF-α(t=3.146,P=0.002 and t=2.973,P=0.002).In addition,they down-regulated the protein levels of TGF-ß1(t=4.657,P=0.001 and t=4.176,P=0.001),p-Smad2/Smad2(t=9.687,P=0.001 and t=6.753,P=0.001) and p-Smad3/Smad3(t=6.477,P=0.002 and t=4.754,P=0.002).Conclusion Tea polyphenols protect rats from heart failure by inhibiting the activation of renin-angiotensin-aldosterone system and TGF-ß1/Smads pathway.

The natural compound from Garcinia bracteata mainly induces GSDME-mediated pyroptosis in esophageal cancer cells.

BACKGROUND: Pyroptosis, an inflammatory form of programmed cell death (PCD), is reported to play important roles in the treatment of tumors. In our previous studies, we found that neobractatin (NBT), a caged prenylxanthone isolated from edible fruits of Garcinia bracteata C. Y. Wu ex Y. H. Li, showed anticancer effects against different cancer cells. However, the effect of NBT on pyroptosis is not well understood. PURPOSE: This study aims to investigate whether and how GSDME-mediated pyroptosis contributes to NBT-induced antitumor effects in esophageal cancer (EC) cells. METHODS: Cell viability assay and colony formation assay were used to determine the anticancer effects of NBT in esophageal cancer cells. Lactate dehydrogenase (LDH) release assay and microscopy imaging were used to detect the main characteristic of pyroptosis. CRISPR-Cas9 knockout and siRNA knockdown were performed to verify the roles of GSDME and caspase-3 in NBT-induced pyroptosis. Flow cytometry was used to measure the reactive oxygen species (ROS) level and cell apoptosis. The changes of related protein level were detected by Western blot. Furthermore, animal experiments were used to verify the in vivo effect of NBT. RESULTS: The results showed that NBT reduced the viability of EC cells mainly through GSDME-mediated pyroptosis. Morphologically, NBT induced cell swelling and formed large bubbles emerging from plasma membrane in wild type EC cells. Furthermore, NBT induced the cleavage of GSDME by activating caspase-3 in EC cells. On the other hand, caspase-3 activated by NBT also induced apoptosis especially at high dosage. Knocking down GSDME switched NBT-induced cell death from mainly pyroptosis to apoptosis in vivo and in vitro. Mechanistic studies indicated that NBT led to accumulation of ROS, which then regulated the phosphorylation of both JNK and MEK/ERK. In the absence of ROS or caspase-3, NBT-induced pyroptosis and apoptosis were completely reversed. Moreover, NBT showed a significant antitumor effect in both the KYSE150 and GSDME knockout KYSE150-/- xenograft models by inducing pyroptosis and apoptosis, respectively. CONCLUSION: Our results indicated that natural compound NBT could induce GSDME-mediated pyroptosis and apoptosis in esophageal cancer cells, making it a potential therapeutic drug in clinical treatment.

YZG-330 regulates the TRPM8 ion channel through the P38 MAPK signaling pathway to reduce body temperature in mice / 药学学报

Preliminary research in our laboratory found that compound YZG-330 can reduce mouse body temperature, which could be blocked by adenosine A1 receptor (A1R) antagonist DPCPX. Based on the downstream signaling pathway of the A1R, the mechanism by which YZG-330 lowers body temperature was further studied. The pharmacodynamics of YZG-330 was evaluated by measuring the rectal temperature; expression of the transient receptor potential (TRP) ion channel, the P38 protein and its phosphorylated form in mouse hypothalamic homogenate were detected by Western blotting. A Ca2+ fluorescent probe, Fluo-3AM, was added to cells to detect the effect of YZG-330 on the Ca2+ content of mouse hypothalamic cells. YZG-330 dose-dependently reduced the body temperature in mice, and the selective P38 inhibitor SB-203580 (20 mg·kg-1, i.p.) significantly inhibited the hypothermic effect of YZG-330. A TRPM8 antagonist 2 (0.1 μg per mouse, i.c.v.) markedly attenuated the hypothermic effect of YZG-330 (0.25 or 1 mg·kg-1, i.p.). YZG-330 (2 mg·kg-1, i.p.) significantly increased the phosphorylation of P38, an effect that could be attenuated by the A1R antagonist DPCPX (5 mg·kg-1, i.g.) in mouse hypothalamus. In addition, YZG-330 also prominently enhanced the expression of TRPM8, which could be blocked by SB-203580; YZG-330 (0.1-10 μmol·L-1) increased intracellular Ca2+ concetration in mouse hypothalamic cells in a dose-dependent manner, and was inhibited by the A1R inhibitor DPCPX (0.5 and 1 μmol·L-1) and TRPM8 antagonist 2 (1 μmol·L-1). In conclusion, YZG-330 exerts its hypothermic effect by activating the A1R to promote the phosphorylation of P38 protein and thereby up-regulating the expression and activity of the TRPM8 ion channel, resulting in increased intracellular Ca2+ concentration to stimulate mouse hypothalamus cells to down-regulate body temperature. All animal experiments were approved by the Ethics Committee of the Institute of Materia Medica, Chinese Academy of Medical Sciences.

[Abdominal acupoint thread embedding therapy based on "brain-intestinal connection" for mild-to-moderate Alzheimer's disease and its effects on serum levels of APP and Aß1-42].

OBJECTIVE: To compare the clinical efficacy of abdominal acupoint thread embedding therapy based on "brain-intestinal connection" combined with donepezil hydrochloride tablets and oral donepezil hydrochloride tablets alone for mild-to-moderate Alzheimer's disease (AD) and observe its effects on amyloid precursor protein (APP) and ß-amyloid protein1-42 (Aß1-42). METHODS: Sixty patients with AD were randomly divided into an observation group (30 cases, 3 cases dropped off) and a control group (30 cases, 3 cases dropped off). The patients in the control group were treated with donepezil hydrochloride tablets (5 mg per day); based on the treatment in the control group, the patients in the observation group were treated with abdominal acupoint thread embedding therapy at Zhongwan (CV 12), Xiawan (CV 10), Huaroumen (ST 24), Wailing (ST 26), Daheng (SP 15), etc., once every 10 days. Both groups were treated for 2 months. The mini-mental state examination (MMSE), Alzheimer's disease assessment scale-cognitive subscale (ADAS-Cog), activity of daily living scale (ADL), neuropsychiatric inventory questionnaire (NPI) as well as the serum levels of APP and Aß1-42 were observed before and after treatment in the two groups. RESULTS: After treatment, the MMSE scores in the two groups were higher than those before treatment (P<0.05), and the ADAS-Cog, ADL and NPI scores were lower than those before treatment (P<0.05). After treatment, the MMSE score in the observation group was higher than that in the control group (P<0.05), and the ADAS-Cog, ADL and NPI scores were lower than those in the control group (P<0.05). After treatment, the serum levels of APP and Aß1-42 were lower than those before treatment (P<0.05), and the serum levels of APP and Aß1-42 in the observation group was lower than those in the control group (P<0.05). CONCLUSION: The abdominal acupoint thread embedding therapy based on the theory of "brain-intestinal connection" combined with donepezil hydrochloride tablets can improve cognitive function, self-care ability of daily life and mental behavior, and reduce the serum levels of APP and Aß1-42 in patients with mild-to-moderate AD, which have superior clinical effect to donepezil hydrochloride tablets alone.

[Effects of natural organic soil amendments on quality of Panax notoginseng].

The effects of four natural organic soil amendments on the quality and pesticide residues of Panax notoginseng were investigated through field experiments and the suitable dosage ratio of each soil amendment was selected to provide a new idea for the pollution-free cultivation of P. notoginseng. The four natural organic soil amendments used in this study were Jishibao, Jihuo, Fudujing, and omnipotent nutrients, which were produced by mixed fermentation of aboveground parts of different plants, biological waste residue, and biochar. During the experiments, only four soil amendments were applied to P. notoginseng instead of any pesticides and fertilizers. The experiment was designed as four factors and three levels. There were three dosage gradients(low, medium, and high) for Jishibao(A), Jihuo(B), Fudujing(C), and omnipotent nutrients(D). When the dosage of one soil amendment changed, the do-sage of the other soil amendments remained medium. There were 10 groups in addition to the soil amendment-free group as control(CK). The results showed that the four soil amendments could significantly improve the growth environment of P. notoginseng and increase the seedling survival rate and saponin content of P. notoginseng. The seedling survival rates of the treatment groups increased by 8.24%-30.05% as compared with the control group. Furthermore, the content of pesticide residues in P. notoginseng was too low to be detected, and that of heavy metals in P. notoginseng was far lower than the specified content in the Chinese Pharmacopoeia(2020). The optimal effect was achieved at medium dosage for all the soil amendments with the highest content of saponins, high seedling survival rate, and significantly reduced heavy metals, such as lead, cadmium, arsenic, and mercury.

Dietary alteration of n-3 and n-6 fatty acids for headache reduction in adults with migraine: randomized controlled trial.

OBJECTIVE: To determine whether dietary interventions that increase n-3 fatty acids with and without reduction in n-6 linoleic acid can alter circulating lipid mediators implicated in headache pathogenesis, and decrease headache in adults with migraine. DESIGN: Three arm, parallel group, randomized, modified double blind, controlled trial. SETTING: Ambulatory, academic medical center in the United States over 16 weeks. PARTICIPANTS: 182 participants (88% women, mean age 38 years) with migraines on 5-20 days per month (67% met criteria for chronic migraine). INTERVENTIONS: Three diets designed with eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), and linoleic acid altered as controlled variables: H3 diet (n=61)-increase EPA+DHA to 1.5 g/day and maintain linoleic acid at around 7% of energy; H3-L6 diet (n=61)-increase n-3 EPA+DHA to 1.5 g/day and decrease linoleic acid to ≤1.8% of energy; control diet (n=60)-maintain EPA+DHA at <150 mg/day and linoleic acid at around 7% of energy. All participants received foods accounting for two thirds of daily food energy and continued usual care. MAIN OUTCOME MEASURES: The primary endpoints (week 16) were the antinociceptive mediator 17-hydroxydocosahexaenoic acid (17-HDHA) in blood and the headache impact test (HIT-6), a six item questionnaire assessing headache impact on quality of life. Headache frequency was assessed daily with an electronic diary. RESULTS: In intention-to-treat analyses (n=182), the H3-L6 and H3 diets increased circulating 17-HDHA (log ng/mL) compared with the control diet (baseline-adjusted mean difference 0.6, 95% confidence interval 0.2 to 0.9; 0.7, 0.4 to 1.1, respectively). The observed improvement in HIT-6 scores in the H3-L6 and H3 groups was not statistically significant (-1.6, -4.2 to 1.0, and -1.5, -4.2 to 1.2, respectively). Compared with the control diet, the H3-L6 and H3 diets decreased total headache hours per day (-1.7, -2.5 to -0.9, and -1.3, -2.1 to -0.5, respectively), moderate to severe headache hours per day (-0.8, -1.2 to -0.4, and -0.7, -1.1 to -0.3, respectively), and headache days per month (-4.0, -5.2 to -2.7, and -2.0, -3.3 to -0.7, respectively). The H3-L6 diet decreased headache days per month more than the H3 diet (-2.0, -3.2 to -0.8), suggesting additional benefit from lowering dietary linoleic acid. The H3-L6 and H3 diets altered n-3 and n-6 fatty acids and several of their nociceptive oxylipin derivatives in plasma, serum, erythrocytes or immune cells, but did not alter classic headache mediators calcitonin gene related peptide and prostaglandin E2. CONCLUSIONS: The H3-L6 and H3 interventions altered bioactive mediators implicated in headache pathogenesis and decreased frequency and severity of headaches, but did not significantly improve quality of life. TRIAL REGISTRATION: ClinicalTrials.gov NCT02012790.

Baihe Jizihuang Tang Ameliorates Chronic Unpredictable Mild Stress-Induced Depression-Like Behavior: Integrating Network Pharmacology and Brain-Gut Axis Evaluation.

Baihe Jizihuang Tang (BHT) is a traditional Chinese medicine (TCM) prescription, which can also be used as a nutritional food with medicinal value. Herein, we aimed to clarify the antidepressive effects and molecular mechanism of BHT. Network pharmacological analysis; chronic unpredictable mild stress (CUMS) rat model assessment; behavioral tests; analysis of hippocampal neurotransmitter levels, hippocampal pathological structure, and hypothalamic-pituitary-adrenal (HPA) axis; western blot analysis; 16s RNA sequencing; ultraperformance liquid chromatography (UPLC)/mass spectrometry (MS); and high-performance liquid chromatography (HPLC)/ultraviolet (UV) analysis were used. We found 8 potentially active components and 12 targets from the database. KEGG analysis suggested that BHT significantly affected BDNF/tyrosine receptor kinase B levels, glutamate binding, synaptic transmission based on neurotransmitter signal, and the response to glucocorticoid signaling pathways. Consistently, 7 chemical components were identified using UPLC/quadrupole time-of-flight/MS; among them, regalosides A, B, C, and E were unique components of lily of TCM, and their content in BHT was significantly different: regaloside A > B > E > C. BHT could nourish hippocampal neurons, affect neurotransmitter metabolism, reduce HPA axis hyperactivity, improve deficits in hippocampal tissue structure, and change depressive behavior. Moreover, BHT regulated BDNF expression in the hippocampus and improved intestinal flora deficits in CUMS rats by changing the content of Bifidobacterium, Rothia, Glutamicibacter, and Lactobacillus at the genus level. Collectively, BHT attenuated CUMS-induced depression-like behavior by regulating BDNF and intestinal flora disorder through the brain-gut axis. Therefore, including BHT in the medication list may constitute a potential strategy for preventing depression.

Tease out the future: How tea research might enable crop breeding for acid soil tolerance.

Unlike most crops, in which soil acidity severely limits productivity, tea (Camellia sinensis) actually prefers acid soils (pH 4.0-5.5). Specifically, tea is very tolerant of acidity-promoted aluminum (Al) toxicity, a major factor that limits the yield of most other crops, and it even requires Al for optimum growth. Understanding tea Al tolerance and Al-stimulatory mechanisms could therefore be fundamental for the future development of crops adapted to acid soils. Here, we summarize the Al-tolerance mechanisms of tea plants, propose possible mechanistic explanations for the stimulation of tea growth by Al based on recent research, and put forward ideas for future crop breeding for acid soils.

Tojapride Reverses Esophageal Epithelial Inflammatory Responses on Reflux Esophagitis Model Rats.

OBJECTIVE: To investigate the mechanism of Tojapride, a Chinese herbal formula extract, on strengthening the barrier function of esophageal epithelium in rats with reflux esophagitis (RE). METHODS: Ten out of 85 SD rats were randomly selected as the sham group (n10), and 75 rats were developed a reflux esophagitis model (RE) by the esophageal and duodenal side-to-side anastomosis. Fifty successful modeling rats were divided into different medicated groups through a random number table including the model, low-, medium-, and high-dose of Tojapride as well as omeprazole groups (n10). Three doses of Tojapride [5.73, 11.46, 22.92 g/(kg•d)] and omeprazole [4.17 mg/(kg•d)] were administrated intragastrically twice daily for 3 weeks. And the rats in the sham and model groups were administered 10 mL/kg distilled water. Gastric fluid was collected and the supernatant was kept to measure for volume, pH value and acidity. Esophageal tissues were isolated to monitor the morphological changes through hematoxylin-eosin (HE) staining, and esophageal epithelial ultrastructure was observed by transmission electron microscopy. The expressions of nuclear factor kappa-light-chain-enhancer of activated B cells p65 (NF-KBp65), κB kinase beta (IKKß), occludin, and zonula occludens-1 (ZO-1) in the esophageal tissues were measured by immunohistochemistry and Western blot, respectively. RESULTS: The gastric pH value in the model group was significantly lower than the sham group (P<0.05). Compared with the model group, gastric pH value in the omeprazole and medium-dose of Tojapride groups were significantly higher (P<0.05). A large area of ulceration was found on the esophageal mucosa from the model rats, while varying degrees of congestion and partially visible erosion was observed in the remaining groups. Remarkable increase in cell gap width and decrease in desmosome count was seen in RE rats and the effect was reversed by Tojapride treatment. Compared with the sham group, the IKKß levels were significantly higher in the model group (P<0.05). However, the IKKß levels were down-regulated after treatment by all doses of Tojapride (P<0.01 or P<0.05). The occluding and ZO-1 levels decreased in the model group compared with the sham group (Ps0.01 or Ps0.05), while both indices were significantly up-regulated in the Tojapride-treated groups (P<0.01 or P<0.05). CONCLUSIONS: Tojapride could improve the pathological conditions of esophageal epithelium in RE rats. The underlying mechanisms may involve in down-regulating the IKKß expression and elevating ZO-1 and occludin expression, thereby alleviating the inflammation of the esophagus and strengthening the barrier function of the esophageal epithelium.

Experimental observation of effects of acupoints, cone numbers and durations of moxibustion with different moxibustion methods on skin surface and inside temperature / 针灸推拿医学（英文版）

Objective: To observe the effects of acupoints, cone numbers and durations of moxibustion with different moxibustion methods on skin surface and inside temperature, and to provide references for the clinical standardization of moxibustion amount. Methods: The 42 big-ear white rabbits were divided into 6 groups according to the random number table method, a 1-cone direct moxibustion group, a 2-cone direct moxibustion group, a 3-cone direct moxibustion group, a 1-cone herbal cake-partitioned moxibustion group, a 2-cone herbal cake-partitioned moxibustion group, and a 3-cone herbal cake-partitioned moxibustion group, with 7 rabbits in each group. Shenque (CV 8), Shenshu (BL 23) and Zusanli (ST 36) were used in each group, but the moxibustion methods, cone numbers and durations of moxibustion were different. Rabbits in each group received moxibustion once every other day for 5 times in total. During the intervention, a thermoelectricity coupled probe and a temperature recorder were used to record the real-time acupoint skin temperature and the temperature at different time points, so as to observe, analyze and process the real-time changes in the temperature difference between the surface and inside of acupoint skin. Results: For herbal cake-partitioned moxibustion, the best temperature for cone changing was (46.38±0.51) ℃ when the highest surface temperature was (49.20±0.52) ℃; the multi-factor comparison of acupoint × cone number × time and acupoint × moxibustion method × time showed that time × acupoint, time × moxibustion method and cone number × acupoint had interactive effects (all P<0.05). Comparing skin temperature differences between different cone numbers at the same acupoint, Shenque (CV 8) on the 1st and the 5th days, Shenshu (BL 23) on the 3rd and the 7th days, Zusanli (ST 36) on the 1st and the 9th days of experiment showed statistically significant differences (all P<0.05). The skin temperature comparison of different moxibustion methods at the same acupoint all had statistical differences (all P<0.05), except for Shenque (CV 8) before moxibustion, Shenshu (BL 23) before moxibustion and on the 5th day; Zusanli (ST 36) only showed statistical differences on the 5th and 7th days (both P<0.05). The skin temperature differences of different acupoints after moxibustion in the 1-cone, 2-cone and 3-cone groups were statistically different (all P<0.05); direct moxibustion and herbal cake-partitioned moxibustion at different acupoints were all statistically different (all P<0.05). Conclusion: Cone changing temperature under the same specifications of herbal cake-partitioned moxibustion was confirmed. Temperature difference between surface and inside of different acupoint skin at the same maximum temperature was significantly different due to the cone numbers and moxibustion methods, which showed the highest at Shenshu (BL 23), the second at Shenque (CV 8), and the lowest at Zusanli (ST 36). The influence of acupoint factor should be considered to determine the quantitative indicators of moxibustion.

Tojapride Reverses Esophageal Epithelial Inflammatory Responses on Reflux Esophagitis Model Rats / 中国结合医学杂志

OBJECTIVE@#To investigate the mechanism of Tojapride, a Chinese herbal formula extract, on strengthening the barrier function of esophageal epithelium in rats with reflux esophagitis (RE).@*METHODS@#Ten out of 85 SD rats were randomly selected as the sham group (n10), and 75 rats were developed a reflux esophagitis model (RE) by the esophageal and duodenal side-to-side anastomosis. Fifty successful modeling rats were divided into different medicated groups through a random number table including the model, low-, medium-, and high-dose of Tojapride as well as omeprazole groups (n10). Three doses of Tojapride [5.73, 11.46, 22.92 g/(kg•d)] and omeprazole [4.17 mg/(kg•d)] were administrated intragastrically twice daily for 3 weeks. And the rats in the sham and model groups were administered 10 mL/kg distilled water. Gastric fluid was collected and the supernatant was kept to measure for volume, pH value and acidity. Esophageal tissues were isolated to monitor the morphological changes through hematoxylin-eosin (HE) staining, and esophageal epithelial ultrastructure was observed by transmission electron microscopy. The expressions of nuclear factor kappa-light-chain-enhancer of activated B cells p65 (NF-KBp65), κB kinase beta (IKKß), occludin, and zonula occludens-1 (ZO-1) in the esophageal tissues were measured by immunohistochemistry and Western blot, respectively.@*RESULTS@#The gastric pH value in the model group was significantly lower than the sham group (P<0.05). Compared with the model group, gastric pH value in the omeprazole and medium-dose of Tojapride groups were significantly higher (P<0.05). A large area of ulceration was found on the esophageal mucosa from the model rats, while varying degrees of congestion and partially visible erosion was observed in the remaining groups. Remarkable increase in cell gap width and decrease in desmosome count was seen in RE rats and the effect was reversed by Tojapride treatment. Compared with the sham group, the IKKß levels were significantly higher in the model group (P<0.05). However, the IKKß levels were down-regulated after treatment by all doses of Tojapride (P<0.01 or P<0.05). The occluding and ZO-1 levels decreased in the model group compared with the sham group (Ps0.01 or Ps0.05), while both indices were significantly up-regulated in the Tojapride-treated groups (P<0.01 or P<0.05).@*CONCLUSIONS@#Tojapride could improve the pathological conditions of esophageal epithelium in RE rats. The underlying mechanisms may involve in down-regulating the IKKß expression and elevating ZO-1 and occludin expression, thereby alleviating the inflammation of the esophagus and strengthening the barrier function of the esophageal epithelium.

Effects of different drying methods on the chemical constituents of Lilium lancifolium Thunb. based on UHPLC-MS analysis and antidepressant activity of the main chemical component regaloside A.

The Lilium lancifolium Thunb. is a herb with multiple functions in both medicine and food in China, and its extracts have shown antidepressant effects. In this study, fresh bulbs of Lilium lancifolium Thunb. were processed to study the effects of different drying processes on changes in its main chemical components. We found that different drying methods can affect the chemical constituents of the herb. Among these components, Regaloside A has been found as the characteristic component. Here, Cell Counting Kit-8 assay, and Western blotting were used to evaluate the neuroprotective antidepressant effects of Regaloside A. The results showed the cell survival rate was improved, the phosphorylation levels of brain-derived neurotrophic factor, tyrosine kinase receptor B, phosphatidylinositol 3 kinase, protein kinase B, and mammalian target of rapamycin were increased after Regaloside A treatment. In general, different drying methods have a significant influence on the chemical composition of the herb, and Regaloside A may be the main chemical component of the herb. It can alleviate the damage of corticosterone in SH-SY5Y cells, and phosphatidylinositol-3-kinase/protein kinase B/mammalian target of rapamycin signaling mediated by brain-derived neurotrophic factor/tyrosine kinase receptor B may play an important role in the neuroprotective antidepressant effects of Regaloside A.

HS-GC-IMS-Based metabonomics study of Baihe Jizihuang Tang in a rat model of chronic unpredictable mild stress.

The aim of this study was to investigate the differences in volatile organic compounds (VOCs) obtained from the feces of a Baihe Jizihuang Tang (BHT)-treated rat depression model. Rats were subjected to chronic unpredictable mild stress (CUMS), and the differences in VOCs were analyzed by headspace-gas chromatography-ion mobility spectrometry (HS-GC-IMS), NIST software, principal component analysis, and orthogonal partial least squares discriminant analysis. Eleven biomarkers were identified on the basis of VOC migration time, and their relative peak intensities were analyzed. A metabonomic model was established using multivariate statistical analysis. The study demonstrated the metabonomics of CUMS rats and the intervention effect of BHT and also highlighted the potential therapeutic effects of the traditional Chinese medicine (TCM) Jingfang for the clinical treatment of complex diseases, which was in line with the holistic and systemic approaches of TCM. This study augments the use of metabonomics based on HS-GC-IMS in research studies. Using this method, there is no need to pre-process samples by extraction or derivatization, and the VOC component of the sample can be detected directly and rapidly. In conclusion, this study establishes a simple, convenient, and fast technique, which can help identify clinical biomarkers for rapid medical diagnosis.

[Method and application of proteomics in study of targets of traditional Chinese medicines].

The study on the targets of traditional Chinese medicine is an important part of researchers using modern scientific language to clarify the mechanism of traditional Chinese medicine. However, the research on the targets of Chinese medicine is full of challenges due to the complexity of active ingredients. As a branch of systems biology, proteomics focus on specific proteins in living organisms from a holistic perspective, which significantly improves the efficiency of targets discovery and has obvious advantages in the research of targets of Chinese medicine. Based on relevant literature and different methods used in targets of Chinese medicine, proteomics can be divided into chemical proteomics, differential proteomics and quantitative proteomics. The applications of the above three methods are illustrated in this paper as well, which will provide new methods and ideas for the study of the mechanism of Chinese medicine in the future.

Koumine Alleviates Lipopolysaccharide-Induced Intestinal Barrier Dysfunction in IPEC-J2 Cells by Regulating Nrf2/NF-κB Pathway.

Gelsemium elegans Benth. (G. elegans), a traditional Chinese medicine, has great potential as an effective growth promoter in animals, however, the mechanism of its actin remains unclear. Here, we evaluated the protective effects of koumine extract from G. elegans against lipopolysaccharide (LPS)-induced intestinal barrier dysfunction in IPEC-J2 cells through alleviation of inflammation and oxidative stress. MTT and LDH assays revealed that koumine significantly reduced LPS cytotoxicity. Transepithelial electrical resistance (TEER) and cell monolayer permeability assays showed that koumine treatment attenuated the LPS-induced intestinal barrier dysfunction with no particularly different effects in tight junction proteins such as ZO-1, claudin-1, and occludin. LPS-triggered inflammatory response was also suppressed by koumine, as evidenced by the downregulated inflammatory factors, including TNF-α, IL-6, IL-1ß, NO, iNOS, and COX-2, which was closely connected with the inhibition of NF-κB pathway for the decrease of phosphorylation of IκBα and NF-κB and nuclear translocation of p-p65. Amount of reactive oxygen species (ROS) and MDA induced by LPS was also reduced by koumine through activation of Nrf2 pathway, and increased in the levels of Nrf2 and HO-1 degradation of keap-1 to promote anti-oxidants, including superoxide dismutase (SOD) and catalase (CAT). To summarize, koumine-reduced the oxidative stress and inflammatory reaction triggered by LPS through regulation of the Nrf2/NF-κB signaling pathway and preventing intestinal barrier dysfunction.

Plasma oxylipins and unesterified precursor fatty acids are altered by DHA supplementation in pregnancy: Can they help predict risk of preterm birth?

Oxidized lipids derived from omega-6 (n-6) and omega-3 (n-3) polyunsaturated fatty acids, collectively known as oxylipins, are bioactive signaling molecules that play diverse roles in human health and disease. Supplementation with n-3 docosahexaenoic acid (DHA) during pregnancy has been reported to decrease the risk of preterm birth in singleton pregnancies, which may be due to effects of DHA supplementation on oxylipins or their precursor n-6 and n-3 fatty acids. There is only limited understanding of the levels and trajectory of changes in plasma oxylipins during pregnancy, effects of DHA supplementation on oxylipins and unesterified fatty acids, and whether and how oxylipins and their unesterified precursor fatty acids influence preterm birth. In the present study we used liquid chromatography-tandem mass spectrometry to profile oxylipins and their precursor fatty acids in the unesterified pool using plasma samples collected from a subset of pregnant Australian women who participated in the ORIP (Omega-3 fats to Reduce the Incidence of Prematurity) study. ORIP is a large randomized controlled trial testing whether daily supplementation with n-3 DHA can reduce the incidence of early preterm birth compared to control. Plasma was collected at study entry (≈pregnancy week 14) and again at ≈week 24, in a subgroup of 48 ORIP participants-12 cases with spontaneous preterm (<37 weeks) birth and 36 matched controls with spontaneous term (≥40 weeks) birth. In the combined preterm and term pregnancies, we observed that in the control group without DHA supplementation unesterified AA and AA-derived oxylipins 12-HETE, 15-HETE and TXB2 declined between weeks 14-24 of pregnancy. Compared to control, DHA supplementation increased unesterified DHA, EPA, and AA, DHA-derived 4-HDHA, 10-HDHA and 19,20-EpDPA, and AA-derived 12-HETE at 24 weeks. In exploratory analysis independent of DHA supplementation, participants with concentrations above the median for 5-lipoxygenase derivatives of AA (5-HETE, Odds Ratio (OR) 8.2; p = 0.014) or DHA (4-HDHA, OR 8.0; p = 0.015) at 14 weeks, or unesterified AA (OR 5.1; p = 0.038) at 24 weeks had higher risk of spontaneous preterm birth. The hypothesis that 5-lipoxygenase-derived oxylipins and unesterified AA could serve as mechanism-based biomarkers predicting spontaneous preterm birth should be evaluated in larger, adequately powered studies.

Pooled analysis of the Xpert MTB/RIF assay for diagnosing tuberculous meningitis.

BACKGROUND: Tuberculous meningitis (TBM) is one of the most serious types of extrapulmonary tuberculosis. However, low sensitivity of culture of cerebrospinal fluid (CSF) increases the difficulty in clinical diagnosis, leading to diagnostic delay, and misdiagnosis. Xpert MTB/RIF assay is a rapid and simple method to detect tuberculosis. However, the efficacy of this technique in diagnosing TBM remains unclear. Therefore, a meta-analysis was conducted to evaluate the diagnostic efficacy of Xpert MTB/RIF for TBM, which may enhance the development of early diagnosis of TBM. METHODS: Relevant studies in the PubMed, Embase, and Web of Science databases were retrieved using the keywords 'Xpert MTB/RIF', 'tuberculous meningitis (TBM)'. The pooled sensitivity, pooled specificity, positive likelihood ratio, negative likelihood ratio, diagnostic odds ratio, summary receiver operator characteristic curve, and area under the curve (AUC) of Xpert MTB/RIF were determined and analyzed. RESULTS: A total of 162 studies were enrolled and only 14 met the criteria for meta-analysis. The overall pooled sensitivity of Xpert MTB/RIF was 63% [95% confidence interval (CI), 59-66%], while the overall pooled specificity was 98.1% (95% CI, 97.5-98.5%). The pooled values of positive likelihood ratio, negative likelihood ratio, and diagnostic odds ratio were 20.91% (12.71-52.82%), 0.40% (0.32-0.50%), and 71.49% (32.64-156.56%), respectively. The AUC was 0.76. CONCLUSIONS: Xpert MTB/RIF exhibited high specificity in diagnosing TBM in CSF samples, but its sensitivity was relatively low. It is necessary to combine other high-sensitive detection methods for the early diagnosis of TBM. Moreover, the centrifugation of CSF samples was found to be beneficial in improving the sensitivity.