双虎清肝颗粒对四氯化碳诱发大鼠肝损伤模型肝组织基因表达谱的影响

目的探讨双虎清肝颗粒对四氯化碳诱发大鼠肝损伤模型肝组织基因表达谱的影响。方法清洁级Sprague—Dawley大鼠60只,随机均分为6组。除正常对照组外,均皮下注射40％四氯化碳8周,各治疗组再给予不同剂量双虎清肝颗粒干预8周;水飞蓟索对照组给予水飞蓟素干预8周,观察干预前后6组大鼠肝脏组织基因表达谱的变化。结果A组大鼠各基因表达无明显变化。四氯化碳干预后B、C1、C2、C3、D组与A组相比,表达下调的基因有：CY相关基因、脂代谢相关基因、蛋白质糖基化相关基因;表达上调的基因有：与细胞周期和细胞凋亡相关的基因、与脂转运相关基因、参与细胞黏附、细胞连接和胶原形成的基因。双虎清肝颗粒干预后C1、C2、C3、D组与B组比较,CY相关基因、蛋白质糖基化相关基因、脂代谢相关基因的表达均上调;双虎清肝颗粒干预前后C1、C2、C3组间比较,干预前上调者干预后下调明显,干预前下调者干预后上调明显,且随着双虎清肝颗粒剂量的增加,基因表达的幅度也增加。双虎清肝颗粒干预后C1、C2、C3组与D组比较,其基因表达的改变和双虎清肝颗粒导致的基因表达改变不同（有下调、有无变化、也有上调）。结论双虎清肝颗粒能够从基因水平减轻四氯化碳所诱发的肝脏损伤,并且量效关系明显。     

Micellar TIA1 with folded RNA binding domains as a model for reversible stress granule formation

TIA1, a protein critical for eukaryotic stress response and stress granule formation, is structurally characterized in full-length form. TIA1 contains three RNA recognition motifs (RRMs) and a C-terminal low-complexity domain, sometimes referred to as a “prion-related domain” or associated with amyloid formation. Under mild conditions, full-length (fl) mouse TIA1 spontaneously oligomerizes to form a metastable colloid-like suspension. RRM2 and RRM3, known to be critical for function, are folded similarly in excised domains and this oligomeric form of apo fl TIA1, based on NMR chemical shifts. By contrast, the termini were not detected by NMR and are unlikely to be amyloid-like. We were able to assign the NMR shifts with the aid of previously assigned solution-state shifts for the RRM2,3 isolated domains and homology modeling. We present a micellar model of fl TIA1 wherein RRM2 and RRM3 are colocalized, ordered, hydrated, and available for nucleotide binding. At the same time, the termini are disordered and phase separated, reminiscent of stress granule substructure or nanoscale liquid droplets.

T cell intracellular antigen-1 (TIA1) has multiple roles within cells, including a critical role in stress granule (SG) formation during eukaryotic cellular stress response (1–3) and translation regulation (4–6). SGs appear in cells exposed to stressors, such as pH, oxidation, and temperature changes, and contain stalled preinitiation RNA–protein complexes. They have been hypothesized to act as a decision point in mRNA processing by helping to guide homeostasis-restoring protein expression or begin apoptosis. Although sometimes associated with misfolded protein aggregates, SG components dissolve and regain function more quickly than other aggregates after the stress is removed (7). TIA1 has three RNA recognition motifs (RRM1, RRM2, RRM3) known to bind RNA with relatively little sequence specificity. TIA1 has a C-terminal low-complexity domain (LCD) enriched with asparagine and glutamine that has been referred to in the literature as a prion-related domain (PRD) because of its sequence similarity to amyloid- or prion-forming proteins. Proteins associated with SGs have also been linked to several human diseases, some characterized as protein misfolding disorders. A mutation within the LCD of TIA1 is the diagnostic marker for Welander distal myopathy (8, 9), and several other TIA1 mutations are linked to amyotrophic lateral sclerosis (10).Despite its importance, little is known about the full-length (fl) or oligomeric form(s) of TIA1 or about the LCD. The RRM domains have been structurally characterized (11–13), giving insight into structure, dynamics, binding, and function. Several excised TIA1-RRM domain constructs were characterized with small-angle scattering and liquid-state NMR; the LCD was excluded from the constructs used in prior published structural studies (11, 13). NMR was used to solve the structure of TIA1-RRM1 (Protein Data Bank [PDB] ID code 5O2V), TIA1-RRM2 bound to the dinucleotide UU-RNA (PDB ID code 5O3J), and TIA1-RRM2,3 (PDB ID code 2MJN). RRM1 appears to contribute little to RNA binding, which may be explained by the negatively charged residues in the RNP1 motif within RRM1. A model of rigid RRM domains with flexible linkers was used to interpret scattering data and show that RRM2 and RRM3 associate more closely with each other and more so after RNA binding than with RRM1. However, the effects of the LCD on the fl structure have remained elusive due to experimental challenges.It has been reported that the LCD of TIA1 can cause phase separation (14). In vivo, phase separation of groups of functionally related, locally concentrated proteins and nucleic acids (14) is believed to lead to the formation of membraneless organelles (biomolecular condensates), such as stress granules (15). Many cellular condensates contain proteins with RNA-binding domains, including Cajal bodies, P bodies, and SGs (15). Misregulation of phase separation has been implicated in several human disease-related functions (8, 9).Many proteins with LCDs also spontaneously partition into separate phases or form gels at high concentrations in vitro, potentially providing a model for in vivo phase separation. The in vitro systems share important properties with the corresponding in vivo systems. Both can undergo transitions and display a continuum of mechanical properties from liquid-like droplets to glassy (16), solid-like particles. Liquid–liquid droplets are typically micrometer, morphologically spherical domains that exhibit liquid-like dynamics in their rapid recovery from photobleaching and solution-state NMR spectra (17). Many liquid droplets or condensates in vitro are metastable and transform over time (16, 18) in a process referred to as hardening or maturation (7, 10, 16, 19). Analogously, membraneless organelles can undergo transitions in vivo during regulated maturation processes (15, 20, 21). Thus, it has been suggested that misfolded, amyloid-like fibrils and aggregates form during maturation of the membraneless organelles (7), suggesting a role for condensates in templating the formation of disease-related fibrils (22). Furthermore, the multiphase in vitro suspensions can be compared to colloids, in that they are homogeneously distributed, stable multiphase suspensions whose formation is controlled by salt concentration, viscogens, and temperature. However useful these analogies are, it is important to note that the in vitro systems are, of course, highly simplified compared to the in vivo situation. The in vivo systems are subject to important biological control over their formation and dissolution and include many other components such as nucleic acids and other proteins.The LCD/PRD of TIA1 has primary sequence similarity to the better-characterized SUP35 prion protein and is also similar to other amyloid-forming proteins (23–25). Atomic force and electron microscopy (EM) have been used to show that TIA1 forms fibers under some conditions (26, 27). Congo red and thioflavin T binding assays have been used to suggest TIA1 forms a cross-β amyloid (26, 28). Sup35 and FUS are proteins with a domain structure and an LCD analogous to TIA1; both have been reported to form amyloids (29). Alternatively, it has been hypothesized that the LCD in multidomain proteins can be unfolded (intrinsically disordered) even in the functional form and that oligomerization might be driven by nonspecific intermolecular interactions between several LCDs on different monomers (7, 30, 31). A dominant hypothesis in the literature has been that the LCD induces disease-related amyloid formation. Many proposed functions of TIA1, such as RNA sequestration into SGs (16), raise the question of whether the RRM domains are folded in the condensates or high-order oligomeric forms.Here we report structural studies of fl apo TIA1 prepared without the use of harsh solvents or denaturants. High-order oligomeric systems such as amyloid fibrils are often challenging systems for traditional structural biology methods. However, solid-state NMR (SSNMR) and EM have been powerful tools for studying these systems. We characterize fl TIA1 with EM and SSNMR to test for the presence of a solid-like phase (fibril), a liquid-like phase (intrinsically disordered protein), or some other structure. We address which domains are folded or ordered, which are solvent exposed, and whether the oligomeric structure is compatible with binding at the RRMs. The answers to these intensely debated questions have consequences for future studies of biomolecular condensates.     

60%四聚乙醛水分散粒剂杀螺效果评价

目的 评价60%四聚乙醛水分散粒剂的室内和现场杀螺效果。方法 按照《农药登记用杀钉螺剂药效试验方法和评价》(NY/ T 1617-2008)要求,采用60%四聚乙醛水分散粒剂(MWG)室内和现场喷洒法灭螺,并与50%氯硝柳胺乙醇胺盐可湿性粉剂(WPN)进行效果比较。结果 使用60%四聚乙醛水分散粒剂室内喷洒法灭螺,1、3 和7 d LC50 值分别为0. 312(0. 271~0. 360)、0. 216(0. 190~0. 245)和0. 177(0. 160~0. 196)g/ m² ,0. 900 g/ m² 浓度7 d 后实验室钉螺死亡率为98. 00%。使用0. 900 g/ m² 浓度现场喷洒7 d,岳阳草洲和江陵县河滩现场钉螺死亡率分别为90. 91%和85. 12%。与50%氯硝柳胺乙醇胺盐可湿性粉剂对照组(1. 000 g/ m² )相比,岳阳草洲现场0. 900、1.800 g/ m² MWG 组钉螺死亡率差异均无统计学意义(χ² = 2. 555、3. 340,P 均>0. 05);江陵县河滩现场1. 800 g/m² MWG 组钉螺死亡率差异无统计学意义(χ² =0. 989,P>0. 05)。结论60%四聚乙醛水分散粒剂室内和现场喷洒试 验使用0. 900 g/ m² 浓度,其药效均达到《农药登记用杀钉螺剂药效试验方法和评价》(NY/ T 1617-2008)规定的登记标准,具有较好的实验室和现场杀螺效果,为新型高效、使用方便的灭螺药物剂型。     

木丹颗粒联合甲钴胺治疗糖尿病周围神经病变的临床观察

目的 观察木丹颗粒联合甲钴胺治疗糖尿病周围神经病变（DPN）的疗效. 方法 选取DPN患者60例,随机分为治疗（Treatment）组和对照（Con）组,每组各30例.Con组口服甲钴胺;Treatment组口服木丹颗粒联合甲钴胺.疗程均为4周.采用总症状评分（TSS）将两组各项指标进行比较. 结果 Treatrent、Con组用药前后TSS评分均改善[（5.60±2.81）vs（2.39±1.90）分,（5.59±2.17）vs（4.08±2.12）分,P均＜0.01].两组治疗后TSS评分差异有统计学意义[（4.08±2.12）vs（2.39±1.90）分,P＜0.01）].结论 与甲钴胺单药治疗相比,木丹颗粒联合甲钴胺可改善DPN患者的临床症状.     

补肾解毒健脾冲剂对慢性乙肝病毒携带者HBeAg、HBVDNA和肝组织学的影响

目的：研讨验证补肾解毒健脾冲剂对慢性乙肝病毒携带者HBV DNA、HBeAg和肝组织学的作用。方法：筛选慢性乙肝病毒携带者200例,以1∶1的比例随机分为治疗组和对照组。治疗组用补肾解毒健脾冲剂治疗,对照组用安慰剂干预,周期为72周。结果：治疗72周后,治疗组患者血清HBV DNA下降﹥1 log10、﹥2 log10及﹥3 log10的患者比例明显高于对照组（P﹤0.005）。治疗组患者血清HBeAg阴转率及转换率明显高于对照组,两组比较差异有统计学意义（P﹤0.05）。治疗组患者治疗前后肝组织的炎症活动度分级及纤维化程度分期相比差异有统计学意义（P﹤0.05）,72周时,与对照组比较,差异有统计学意义（P﹤0.05）。结论：补肾解毒健脾冲剂对慢性乙肝病毒携带者的HBeAg血清学转换及HBV DNA阴转作用明显,对肝脏组织炎症活动度及纤维化程度也有改善作用,安全性好,未见明显副作用。     

化滞柔肝颗粒对酒精联合脂多糖诱导的酒精性肝炎小鼠的保护作用*

目的 观察化滞柔肝颗粒对酒精联合脂多糖诱导的酒精性肝炎小鼠的保护作用。方法 将80只ICR小鼠随机分成正常组、模型组、小、中、大剂量化滞柔肝组和护肝片处理组,通过灌胃给予56%北京红星二锅头（12 ml?kg-1.d-1）和脂多糖（5 mg?kg-1,2次/w）腹腔注射,建立酒精性肝炎小鼠模型,同时给予相应的药物灌胃,连续10 w。在末次给药后,解剖动物,取血清和肝组织,进行相应的检查。结果 在实验10 w末,模型组肝组织肝细胞水肿,胞浆疏松,其肝质量指数、血清ALT和AST水平分别为 （5.77±0.67） %、（82.22±6.20） U/L和 （93.43±17.30） U/L,均显著高于正常组[分别为（4.44±0.42） %、（35.83±3.84） U/L和 （66.43±5.14） U/L,P<0.05];大剂量化滞柔肝组肝质量指数为（5.24±0.36）%,显著低于模型组 [（5.77±0.67） %,P<0.05],小、中、大剂量化滞柔肝颗粒组和护肝片组ALT和AST水平均显著低于模型组（P<0.001）;小、中、大剂量化滞柔肝颗粒组和护肝片组的CK分别为（118.93±10.15） U/L、（102.33±8.07） U/L、（119.45±19.26） U/L和（104.00±8.15） U/L,均显著低于模型组[（227.50±50.10） U/L,P<0.001];小、中、大剂量化滞柔肝组和护肝片组血清TBIL水平均显著低于模型组（P<0.01）;各试验组之间血脂水平均无明显变化。结论 化滞柔肝颗粒对酒精联合脂多糖诱导的酒精性肝炎小鼠有保护作用,为化滞柔肝颗粒将来用于临床治疗酒精性脂肪肝提供了试验支持。     

Synaptic clusters function as odor operators in the olfactory bulb

How the olfactory bulb organizes and processes odor inputs through fundamental operations of its microcircuits is largely unknown. To gain new insight we focus on odor-activated synaptic clusters related to individual glomeruli, which we call glomerular units. Using a 3D model of mitral and granule cell interactions supported by experimental findings, combined with a matrix-based representation of glomerular operations, we identify the mechanisms for forming one or more glomerular units in response to a given odor, how and to what extent the glomerular units interfere or interact with each other during learning, their computational role within the olfactory bulb microcircuit, and how their actions can be formalized into a theoretical framework in which the olfactory bulb can be considered to contain “odor operators” unique to each individual. The results provide new and specific theoretical and experimentally testable predictions.The organization of olfactory bulb network elements and their synaptic connectivity has evolved to subserve special computational functions needed for odor detection and recognition (1–5). Key to this organization are the olfactory glomeruli, collecting input from olfactory receptor neuron subsets. These connect to the dendrites of mitral, tufted, and periglomerular cells, and the mitral and tufted cells in turn connect to granule cells. We term these interconnected cells a cluster, and a cluster related to a given glomerulus is a glomerular unit (GU), often visualized as a column of granule cell bodies located below a glomerulus (6, 7). The existence of such GUs has also been suggested from 2-deoxyglucose (8) and voltage-sensitive dye studies (9).Understanding the neural basis of odor processing therefore requires understanding the computational functions and role of GUs. These issues, which are difficult or impossible in experiments, can be conveniently explored using realistic computational models, provided they are able to explain and reproduce crucial experimental findings on glomerular clusters or units.Analyzing synaptic interactions between cells with overlapping dendrites requires modeling in real 3D space. Scaling up to the network level further requires scaling up realistic structural and functional properties to many thousands of cells (10). Building on this unique approach, we show that this model generates columnar clusters of cells related to individual glomeruli, as in the experiments, and further demonstrates mechanisms of odor processing within and between the GUs. Finally, interpreting this network activity requires a theoretical framework, incorporating distributed activated glomeruli within the global network, for which we introduce the concept of the odor operator. The results provide a basis for extension to the glomerular level on the one hand and interactions with olfactory cortex on the other.     

Control of cerebellar granule cell output by sensory-evoked Golgi cell inhibition

Classical feed-forward inhibition involves an excitation–inhibition sequence that enhances the temporal precision of neuronal responses by narrowing the window for synaptic integration. In the input layer of the cerebellum, feed-forward inhibition is thought to preserve the temporal fidelity of granule cell spikes during mossy fiber stimulation. Although this classical feed-forward inhibitory circuit has been demonstrated in vitro, the extent to which inhibition shapes granule cell sensory responses in vivo remains unresolved. Here we combined whole-cell patch-clamp recordings in vivo and dynamic clamp recordings in vitro to directly assess the impact of Golgi cell inhibition on sensory information transmission in the granule cell layer of the cerebellum. We show that the majority of granule cells in Crus II of the cerebrocerebellum receive sensory-evoked phasic and spillover inhibition prior to mossy fiber excitation. This preceding inhibition reduces granule cell excitability and sensory-evoked spike precision, but enhances sensory response reproducibility across the granule cell population. Our findings suggest that neighboring granule cells and Golgi cells can receive segregated and functionally distinct mossy fiber inputs, enabling Golgi cells to regulate the size and reproducibility of sensory responses.Classical feed-forward inhibition (FFI) involves a sequence of excitation rapidly terminated by inhibition. This temporal sequence narrows the time window for synaptic integration and enforces precise spike timing (1–7). FFI is thought to be important for regulating the temporal fidelity of spike responses in many neural systems, including the motor system, where rapid and adaptable changes in muscle activity are essential for coordinated motor control (8–10). The cerebellum plays a central role in fine sculpting of movements, and damage to the cerebellum produces severe motor deficits, most notably enhanced temporal variability of voluntary movements (11, 12). These findings suggest that cerebellar circuits have the ability to preserve precise timing information during behavior (5, 6, 13), and in vitro studies have shown that feed-forward inhibitory networks in the input layer of the cerebellum provide a mechanism for maintaining the temporal fidelity of information transmission (6, 14, 15).Synaptic inhibition in the granule cell layer is generated by Golgi cells, GABAergic interneurons that provide direct inhibitory input to granule cells (6, 15–17). The prevailing view is that, when mossy fibers are activated, granule cells receive both monosynaptic excitation and disynaptic FFI from Golgi cells, providing temporally precise inhibitory input that narrows the window for the temporal summation of discrete mossy fiber inputs (6, 14, 18). This classical excitation–inhibition sequence forms the basis of a variety of contemporary cerebellar models (7, 9, 18, 19). However, the exact temporal relationship between sensory-evoked excitation and inhibition in granule cells has never been determined in vivo. Here, we combined in vivo whole-cell voltage-clamp recordings from granule cells and in vitro dynamic clamp experiments to investigate both the temporal dynamics of Golgi-cell–mediated inhibition and its importance for shaping sensory responses in the input layer of the cerebellum.     

利肝克纤冲剂抗肝纤维化的临床研究

目的：观察中药利肝克纤冲剂在抗肝纤维化方面的疗效。方法：选取90例肝纤维化患者随机分为两组，治疗组60例，对照组30例，治疗6个月。观察治疗前后两组患者的症状、体征、肝功能、肝纤维化指标及B超影像学的变化，评价中药利肝克纤冲剂抗肝纤维化的疗效。结果：治疗组总有效率为86．67％，对照组总有效率为43．33％，两组比较，差异具有显著性意义（P〈0．01）。结论：利肝克纤冲剂可明显改善肝纤维化患者的症状、体征、肝功能、血清肝纤维化4项指标（HA、LN、PCⅢ及Ⅳ-c）及B超影像学表现，具有明显抗肝纤维化疗效。