ObjectiveThis study aimed to explore shared genetic etiology and the causality between smoking status and type 2 diabetes (T2D), cardiovascular diseases (CVDs), and related metabolic traits.MethodsUsing summary statistics from publicly available genome-wide association studies (GWASs), we estimated genetic correlations between smoking status and T2D, 6 major CVDs, and 8 related metabolic traits with linkage disequilibrium score regression (LDSC) analysis; identified shared genetic loci with large-scale genome-wide cross-trait meta-analysis; explored potential shared biological mechanisms with a series of post-GWAS analyses; and determined causality with Mendelian randomization (MR).ResultsWe found significant positive genetic associations with smoking status for T2D (Rg = 0.170, p = 9.39 × 10−22), coronary artery disease (CAD) (Rg = 0.234, p = 1.96 × 10−27), myocardial infarction (MI) (Rg = 0.226, p = 1.08 × 10−17), and heart failure (HF) (Rg = 0.276, p = 8.43 × 10−20). Cross-trait meta-analysis and transcriptome-wide association analysis of smoking status identified 210 loci (32 novel loci) and 354 gene–tissue pairs jointly associated with T2D, 63 loci (12 novel loci) and 37 gene–tissue pairs with CAD, 38 loci (6 novel loci) and 17 gene–tissue pairs with MI, and 28 loci (3 novel loci) and one gene–tissue pair with HF. The shared loci were enriched in the exo-/endocrine, cardiovascular, nervous, digestive, and genital systems. Furthermore, we observed that smoking status was causally related to a higher risk of T2D (β = 0.385, p = 3.31 × 10−3), CAD (β = 0.670, p = 7.86 × 10−11), MI (β = 0.725, p = 2.32 × 10−9), and HF (β = 0.520, p = 1.53 × 10−6).ConclusionsOur findings provide strong evidence on shared genetic etiology and causal associations between smoking status and T2D, CAD, MI, and HF, underscoring the potential shared biological mechanisms underlying the link between smoking and T2D and CVDs. This work opens up a new way of more effective and timely prevention of smoking-related T2D and CVDs.

Background Microgravity leads to declined cardiac function and remodeling which could cause pathological problems in the postflight period. Time-restricted feeding (TRF), as a form of intermittent fasting, is an emerging approach to improve health, including cardiometabolic metrics, in animals and humans, but whether it could benefit cardiac performance of astronauts in long-term spaceflight, remains unknown. Aims In this study, we investigated the effects of TRF (food access limited to 8 hours every day) on cardiac metabolism and performance under simulated microgravity. Methods and Results Male rats under ad libitum feeding or TRF were subjected to hindlimb unloading (HU) to simulate microgravity. After 6-week intervention, cardiac function was assessed by two-dimensional speckle tracking echocardiography. HU for 6 weeks led to left ventricular dyssynchrony and declined cardiac function. HU also resulted in cardiac atrophy and collagen deposition. All these were ameliorated by TRF except for cardiac atrophy. Futhermore, metabolites in the heart and blood were assessed by liquid chromotography-tandom mass spectrometry. HU suppressed cardiac glucose utilization and glutaminolysis, indicating a suppressed cardiac flexibility, which can also be restored by TRF. Significance Our data demonstrate that TRF improves cardiac glucose and glutamine metabolism, ameliorates cardiac dysfunction and remodeling in the rats exposed to simulated microgravity. Our study shed new lights on the cardiometabolic regulation of TRF and suggest TRF as a potential countermeasure for cardiac protection in long-term spaceflight.

目的 观察太子参多糖(总多糖)、葛根淫羊藿总黄酮(总黄酮)及其复合物对小鼠脑蛋白质组学的影响,探讨其神经细胞网络调控的潜在机理.方法 给正常小鼠分别灌胃太子参多糖(400 mg·kg-1)、葛根淫羊藿总黄酮(400 mg·kg-1)及其二者复合物(800 mg·kg-1),连续给药7天,第7天最后1次给药后1 h,处死动物,取脑.提取蛋白,利用LC/MS-MS测试蛋白表达的变化.通过DAVID数据库分析变化蛋白的功能及信号通路,STRING数据库分析变化蛋白的相互作用及其网络关系.结果 在所测试的5567个蛋白中,以上调蛋白占比较高,其中主要是线粒体的基因受到调控.总多糖、总黄酮及其复合物都上调的蛋白有8个,作用于线粒体的不同环节,提示该复合物抗抑郁作用的靶点主要在线粒体.太子参多糖的调控线粒体相关蛋白的作用揭示了其潜在的神经精神方面的药理作用.除此,还发现各有效部位调控蛋白的数量明显多于复合物,提示药物复合作用的复杂性.结论 总多糖、总黄酮及其复合物可以作用于脑神经元,并且主要以线粒体为主要靶点.该研究为相关中药调控神经精神功能活动提供了重要的蛋白线索及其实验依据.

Berberine (BBR) has a variety of pharmacological activities.Studies have reported that BBR not only reduces heat stress-induced fever but also inhibits lower body temperatures due to cold stress.Heat stress can be reduced via BBR treatment,which antagonizes HSP70-TNFα to regulate the body temperature alteration.In cold stress,however,the molecular mechanism of BBR-induced inhibition of hypothermia remains unclear.Therefore,we studied whether BBR promoted uncoupling protein 1 (UCP1,a crucial protein of thermogenesis) expression and its mechanism under cold stress.Wild type mice and Ucp1-/ mice were used for the in vivo experiments,and primary brown adipocytes and brown adipocytes HIB-1B were used for the in vitro studies.The cold stress was set at 4 ℃.The results showed that at 4 ℃C,the body temperature of mice was decreased.BBR effectively inhibited this hypothermia.Simultaneously,Ucp1 expression in brown adipose tissue (BAT) cells was significantly increased,and BBR promoted Ucp1 expression.However,in Ucp1-knockout mice,the effect of BBR on hypothermia disappeared during cold stress,indicating that the main target for BBR regulation of body temperature was Ucp1.Further studies showed that the transcriptional response element NFE2 (nuclear factor erythroid-derived 2) in the upstream of the Ucp1 promoter region contributed to the positive regulatory role on Ucp1 expression at lower temperature.BBR could bind to the sequence of NFE2 response element in a temperature-dependent manner.Increased affinity of BBR binding to NFE2 response element in cold stress significantly strengthened and enhanced the expression of Ucp1.This work was important for understanding the role of BBR on thermogenesis in BAT,body temperature regulation and temperature tolerance under cold conditions.

Transient Receptor Potential Melastatin-8 (TRPM8) reportedly plays a fundamental role in a variety of processes including cold sensation, thermoregulation, pain transduction and tumorigenesis. However, the role of TRPM8 in inflammation under cold conditions is not well known. Since cooling allows the convergence of primary injury and injury-induced inflammation, we hypothesized that the mechanism of the protective effects of cooling might be related to TRPM8. We therefore investigated the involvement of TRPM8 activation in the regulation of inflammatory cytokines. The results showed that TRPM8 expression in the mouse hypothalamus was upregulated when the ambient temperature decreased; simultaneously, tumor necrosis factor-alpha (TNFα) was downregulated. The inhibitory effect of TRPM8 on TNFα was mediated by nuclear factor kappa B (NFκB). Specifically, cold stress stimulated the expression of TRPM8, which promoted the interaction of TRPM8 and NFκB, thereby suppressing NFκB nuclear localization. This suppression consequently led to the inhibition of TNFα gene transcription. The present data suggest a possible theoretical foundation for the anti-inflammatory role of TRPM8 activation, providing an experimental basis that could contribute to the advancement of cooling therapy for trauma patients.

After being studied for approximately a century, berberine (BBR) has been found to act on various targets and pathways. A great challenge in the pharmacological analysis of BBR at present is to identify which target(s) plays a decisive role. In the study described herein, a rescue experiment was designed to show the important role of mitochondria in BBR activity. A toxic dose of BBR was applied to inhibit cell proliferation and mitochondrial activity, then α-ketobutyrate (AKB), an analogue of pyruvate that serves only as an electron receptor of NADH, was proven to partially restore cell proliferation. However, mitochondrial morphology damage and TCA cycle suppression were not recovered by AKB. As the AKB just help to regenerate NAD+, which is make up for part function of mitochondrial, the recovered cell proliferation stands for the contribution of mitochondria to the activity of BBR. Our results also indicate that BBR suppresses tumour growth and reduces energy charge and mitochondrial DNA (mtDNA) copy number in a HepG2 xenograft model. In summary, our study suggests that mitochondria play an important role in BBR activity regarding tumour cell proliferation and metabolism.

Pomegranate leaf (PGL) has a definite role in regulating lipid metabolism. However, pharmacokinetic results show the main active ingredient, ellagic acid, in PGL has lower oral bioavailability, suggesting that the lipid-lowering effect of PGL may act through inhibiting lipid absorption in the small intestine. Our results demonstrated that pomegranate leaf and its main active ingredients (i.e., ellagic acid, gallic acid, pyrogallic acid and tannic acid) were capable of inhibiting pancreatic lipase activity in vitro. In computational molecular docking, the four ingredients had good affinity for pancreatic lipase. Acute lipid overload experiments showed that a large dosage of PGL significantly reduced serum total cholesterol (TG) and triglycerides (TC) levels in addition to inhibiting intestinal lipase activity, which demonstrated that PGL could inhibit lipase activity and reduce the absorption of lipids. We also found that PGL could reverse the reduced tight-junction protein expression due to intestinal lipid overload, promote Occludin and Claudin4 expression in the small intestine, and enhance the intestinal mucosal barrier. In conclusion, we demonstrated that PGL can inhibit lipid absorption and reduce blood TG and TC by targeting pancreatic lipase, promoting tight-junction protein expression and thereby preventing intestinal mucosa damage from an overload of lipids in the intestine.

There have been many reports on berberine (BBR) effect of the inhibition on gut bacteria,but more from the protein level.In view of the preference of BBR for DNA binding,we here investigated the expression of BBR from the transcriptional expression level of the gene.The results showed that BBR had a higher affinity for UP element of Escherichia coli (E.coli) gene,and the transcription initiation region of this element contained TATA base sequence.The expression of genes sulA,recA and 16S which contain the genes of the UP element regulatory elements in the upstream of the promoter could be suppressed by BBR,and the expression of lpxC,secG and mutT which did not contain the genes of the UP element regulatory elements in the upstream of the promoter could not be inhibited by BBR.It is shown that the TATA sequence is the target of BBR.This result provides a new perspective for exploring the effect of BBR's inhibition of microbiota from gene transcription.

OBJECTIVE:To explore the anti-nociceptive effect of patchouli alcohol (PA), the essential oil isolated from Pogostemon cablin (Blanco) Bent, and determine the mechanism in molecular levels.METHODS:The acetic acid-induced writhing test and formalin-induced plantar injection test in mice were employed to confirm the effect in vivo. Intracellular calcium ion was imaged to verify PA on mu-opioid receptor (MOR). Cyclooxygenase 2 (COX2) and MOR of mouse brain were expressed for determination of PA's target. Cellular experiments were carried out to find out COX2 and MOR expression induced by PA.RESULTS:PA significantly reduced latency period of visceral pain and writhing induced by acetic acid saline solution (P<0.01) and allodynia after intra-plantar formalin (P<0.01) in mice. PA also up-regulated COX2 mRNA and protein (P<0.05) with a down-regulation of MOR (P<0.05) both in in vivo and in vitro experiments, which devote to the analgesic effect of PA. A decrease in the intracellular calcium level (P<0.05) induced by PA may play an important role in its anti-nociceptive effect. PA showed the characters of enhancing the MOR expression and reducing the intracellular calcium ion similar to opioid effect.CONCLUSIONS:Both COX2 and MOR are involved in the mechanism of PA's anti-nociceptive effect, and the up-regulation of the receptor expression and the inhibition of intracellular calcium are a new perspective to PA's effect on MOR.

Heat stress can stimulate an increase in body temperature, which is correlated with increased expression of heat shock protein 70 (HSP70) and tumor necrosis factor α (TNFα). The exact mechanism underlying the HSP70 and TNFα induction is unclear. Berberine (BBR) can significantly inhibit the temperature rise caused by heat stress, but the mechanism responsible for the BBR effect on HSP70 and TNFα signaling has not been investigated. The aim of the present study was to explore the relationship between the expression of HSP70 and TNFα and the effects of BBR under heat conditions, using in vivo and in vitro models. The expression levels of HSP70 and TNFα were determined using RT-PCR and Western blotting analyses. The results showed that the levels of HSP70 and TNFα were up-regulated under heat conditions (40 °C). HSP70 acted as a chaperone to maintain TNFα homeostasis with rising the temperature, but knockdown of HSP70 could not down-regulate the level of TNFα. Furthermore, TNFα could not influence the expression of HSP70 under normal and heat conditions. BBR targeted both HSP70 and TNFα by suppressing their gene transcription, thereby decreasing body temperature under heat conditions. In conclusion, BBR has a potential to be developed as a therapeutic strategy for suppressing the thermal effects in hot environments.

目的:石榴叶总鞣质（PLT）具有降低高糖高脂模型动物以及单纯性肥胖动物模型血中三酰甘油（TG）的作用。方法:为深入研究其降脂作用机理,在给予小鼠一定量的PLT后不同时间一次性灌胃橄榄油,测定血中TG浓度变化与时间的关系。结果:实验结果显示,提前0.5 h灌胃PLT可以显著降低正常小鼠和肥胖小鼠口服橄榄油所致的血中TG水平上升,但是对于腹腔注射橄榄油引起的TG升高没有作用,鞣酸可以抑制口服和灌胃橄榄油引起的TG升高;提前2 h给予PLT对于复合橄榄油引起的TG的升高作用不明显,但是可以降低血中总胆固醇（TC）和血糖的浓度,鞣酸可以降低复合橄榄油引起的TG、TC和血糖的升高。结论:总之,PLT可以显著减少肠道脂质的吸收,起到降脂减肥的作用。PLT中的鞣酸为其主要有效成分。

The purpose of this study is to investigate the effects of carpal tunnel syndrome on hand intrinsic muscle activities using surface electromyogram (sEMG) during precision grip. Six CTS patients and six age-and gender-matched healthy volunteers participated in the experiment. The subjects were instructed to stably grip an instrumented apparatus with their thumb and index finger for 80 s. The sEMG signals were recorded from the abductor pollicis brevis (APB) and first dorsal interosseous (FDI). Visual feedback was provided for the first 20-40 s and removed for the remaining 50-70 s during the precision grip. Results showed that the root mean squares (RMS) of sEMG in CTS significantly increased after removing visual feedback. In contrast of the controls, the CTS led to an increase in the RMS without visual feedback. And the median frequency (MF) was lower than the healthy in APB but higher in the left hand FDI on both feedback conditions. However, CTS did not affect the coherence of the two intrinsic muscles-the APB and FDI. There was a weak coherence in both the CTS and control groups. This study revealed that the sensorimotor deficits associated with CTS could alter the intrinsic muscle activities for precision grip.

Brazilein is reported to have immunosuppressive effect on cardiovascular and cerebral-vascular diseases. The essential roles of innate immunity in cerebral ischemia are increasingly identified, but no studies concerning the influence of brazilein on the innate immunity receptors have been reported. The present study was designed to investigate the regulation of NOD2 (Nucleotide-binding oligomerization domain-containing protein 2) by brazilein for its protection of neuron in cerebral ischemia in vivo and oxygen-glucose deprivation in vitro. The results showed that brazilein could reverse the elevated expression of NOD2 and TNFα (tumor necrosis factor alpha) elicited by cerebral ischemia and reperfusion. This reduction could also be detected in normal mice and C17.2 cells, indicating that this suppressive effect of brazilein was correlated with NOD2. The results from GFP reporter plasmid assay suggested brazilein inhibited NOD2 gene transcription. In conclusion, brazilein could attenuate NOD2 and TNFα expression in cerebral ischemia and NOD2 may be one possible target of brazilein for its immune suppressive effect in neuro-inflammation.

This study investigated effects of carpal tunnel syndrome (CTS) on digit flexibility and range of motion. Twelve healthy, right-handed subjects attended the experiment (6 patients with CTS and 6 healthy controls). The digit flexibility was tested through a Purdue Pegboard Test and the digit's range of motion was tested in process of thumb opposition and thumb circumduction movements. Kinematic signals during thumb movement was recorded using an electromagnetic motion capture system. Additional hand sensorimotor function was assessed using finger sensitivity test, grip strength test and pinch strength tests). The path lengths and interphalangeal (IP) joint angles during thumb opposition and circumduction movements were calculated. Results showed that the path lengths and the angle of the IP joint were significantly lower in the CTS patients compared with controls. At the same time we also found that control group has higher sensitivity than CTS group. There is no obvious change trend for the pinch strength test and grip strength test.

中药小分子是一类从天然产物中分离得到的具有特定生物活性的小分子化合物。按照现代药物发现类型的划分标准,中药小分子的发现主要依靠药效筛选。基于化合物的药学特性,构建和优化高精度、高通量新型药物筛选系统,需要研究人员不断将新的生物学技术和理念应用于中药小分子药理作用机制的准确认识。本综述将重点讨论小分子药理机制研究中对药物作用靶点的认识,为中药小分子研究思路提供借鉴。同时,本文还介绍了一些利用前沿生物学技术研究小分子药理的实例,最后讨论小分子药理机制研究对现代药物发现的意义。

Caenorhabditis elegans (C.elegans), has developed into an useful model for biomedical research, particularly in the targets of novel drugs and the related mechanism.C.elegans, combining with cost-effective cultivation, simple biological structure, make for high-throughput screenings in vivo model.In this review, we explore the feasibility of C.elegans in drug activity evaluation of complex systems.Because of the body independence of C.elegans, it provided good platform for the efficacy and mechanism of research on new drugs as well as Chinese herbal extract in vivo, showing a good prospect.

This study was aimed to observe the effect ofpomegranate leaf tennis and Ananas coumosus leaf phenols on the growth of lactating mice through breast milk. Intragastric administration of different doses of drug was given to maternal mice. The general condition of newborn mice, body weight, tail length and organ coefficient of liver-related factors regulating the expression of lipid metabolism were observed. The results showed that both pomegranate leaf tennis and A. coumosus leaf phenols can obviously affect the growth and development of newborn mice through breast milk. And the effect of pomegranate leaf tennis was stronger than A. coumosus leaf phenols. Meanwhile, it obviously downregulated the expression of liver-related lipid metabolism enzymes in newborn mice. It was concluded that pomegranate leaf tennis and A. coumosus leaf phenols can affect the growth and development of newborn mice through breast milk. Its effect was related to the influence of lipid metabolism enzyme in the liver. Attention should be paid on taking this medication during lactation. Its clinical significance still required in-depth study.

This study was aimed to observe effect of pomegranate leaf tannin and ellagic acid on primary rat adipocyte differentiation and lipid metabolism-related factor expression. Primary rat preadipocyte was in vitro cultured to observe the effect of pomegranate leaf tannin and ellagic acid on lipid fat cells as well as mRNA expression of its related factor. The results showed that pomegranate leaf tannin and ellagic acid had obvious inhibition effect on fat formation in fat cells. It had certain inhibition effect on activities of lipoprotein lipase (LPL) and glucose-3-phosphate dehydrogenase (GPDH). It promoted fat decomposition and reduced intracellular lipid content. It upregulated PPARγ and fatty acid-binding protein (aP2). It downregulated obese (ob) gene level. It was concluded that pomegranate leaf tannin can inhibit fat generation of fat cells and promote fat metabolism. Ellagic acid was its main active ingredient, which had the same effect as pomegranate leaf tannin.

The purpose of the present study is to confirm the protective effect of berberine (BBR) on gastrointestinal injury caused by acute heavy alcohol exposure, an effect that has not been reported previously. Our research details how BBR protects against gastrointestinal injuries from acute alcohol exposure using both in vivo and in vitro experiments. Acute high alcohol concentrations lead to obvious damage to the gastrointestinal mucosa, resulting in necrosis of the intestinal mucosa. Oral administration of BBR was able to significantly reduce this alcohol-induced damage, inhibit increases of alcohol-induced TNFα and IL-1β expression in gastrointestinal mucosa as well as their upstream signals TLR2 and TLR4, and regulate cytokines that modulate tight junctions. Alcohol consumption is a popular human social behavior worldwide, and the present study reports a comprehensive mechanism by which BBR protects against gastrointestinal injuries from alcohol stress, providing people with a novel application of BBR.