Chinese Medicine Huzhen Tongfeng Formula Effectively Attenuates Gouty Arthritis by Inhibiting Arachidonic Acid Metabolism and Inflammatory Mediators.

The Chinese herbal medicine, Huzhen Tongfeng Formula (HZTF), derived from traditional Chinese medicine (TCM) practice, has recognized therapeutic benefits for gouty arthritis (GA). HZTF is currently in the late stage of approval process as a new anti-GA drug application. However, the underlying mechanism of HZTF as an antigout medication is unclear. In this study, we combined network pharmacology and experimental validation approaches to elucidate the mechanism of action of HZTF. First, the relative drug-disease target networks were constructed and analyzed for pathway enrichment. Potential pathways were then validated by in vitro and in vivo experiments. We found that 34 compounds from HZTF matched 181 potential drug targets. Topology analysis revealed 77 core targets of HZTF, which were highly related to gout, following screening of KEGG pathway enrichment. Further analysis demonstrated that the arachidonic acid metabolic pathway was the most relevant pathway involved in the mechanism of HZTF. Validation experiments showed that HZTF significantly inhibited the inflammatory cell infiltration into gouty joints, improved the swelling of affected joints, and increased the pain threshold. HZTF significantly reduced the transcription and production of various cytokines and inflammatory mediators in vitro. In particular, cyclooxygenase (COX)-1, COX-2, and 5-lipoxygenase were simultaneously downregulated. In conclusion, our study suggests that the antigout mechanism of HZTF is associated with the inhibition of the arachidonic acid pathway, resulting in the suppression of inflammatory cytokines and mediators. These findings extend our understanding of the pharmacological action of HZTF, rationalizing the application HZTF as an effective herbal therapy for GA.

Berberine-Loaded Nanostructured Lipid Carriers Enhance the Treatment of Ulcerative Colitis.

PURPOSE: Berberine (BBR), a major ingredient extracted from Coptis chinensis, is a natural drug with limited oral bioavailability. We developed nanostructured lipid carriers (NLCs) as a delivery system for enhanced anti-inflammatory activity of BBR against ulcerative colitis (UC). METHODS: BBR-loaded nanostructured lipid carriers (BBR-NLCs) prepared via high-pressure homogenization were evaluated for particle size, zeta potential, drug entrapment efficiency, drug loading, drug release, toxicity, and cellular uptake. The anti-UC activities of free and encapsulated BBR were evaluated in a DSS-induced acute model of UC in mice. RESULTS: Spherical BBR-NLCs were prepared with a particle size of 63.96± 0.31 nm, a zeta potential of +3.16 ± 0.05 mV, an entrapment efficiency of 101.97±6.34%, and a drug loading of 6.00±0.09%. BBR-NLCs showed excellent biocompatibility in vivo. Cellular uptake experiments showed that BBR-NLCs improved uptake of BBR by RAW 264.7 cells and Caco-2 cells. Oral administration of BBR-NLCs significantly alleviated colitis symptoms (DAI, colon length, spleen swelling, MPO activity) through inhibition of NF-κB nuclear translocation, decreased expression of pro-inflammatory cytokines (IL-1ß, IL-6, MMP-9, CX3CR1, COX-2, TERT), and increased expression of the tight junction protein ZO-1. CONCLUSION: BBR-loaded NLCs improved colitis symptoms, which suggested that this may be a novel formulation for treatment of UC.

Sanhuang Shu'ai decoction alleviates DSS-induced ulcerative colitis via regulation of gut microbiota, inflammatory mediators and cytokines.

BACKGROUND: Sanhuangshu'ai decoction (SH), a traditional Chinese medicine (TCM) prescription, has been safely used to treat diarrhea, dysentery and other inflammatory diseases with little side effect and low cost for thousands of years. However, its mechanism remains elusive. This study was designed to investigate the anti-ulcerative colitis (UC) activity of SH and mechanism by detecting its anti-inflammatory, anti-oxidative, and intervention effects of intestinal flora with the dextran sodium sulfate (DSS)-induced colitis mice. METHODS: The DSS-induced colitis mice was orally administered SH for 1 week with 0.8 or 1.6 g kg-1 d-1 dosage. A clinical disease activity score was evaluated daily. The colonic tissues of the mice were collected and prepared to detect its anti-inflammatory, anti-oxidative, intervention effects of intestinal flora and hydrogen peroxide(H2O2) in vivo, cytotoxicity and ROS influencing effects in vitro. Histological colitis severity and expression of cytokines were also determined. RESULTS: Oral administration of SH significantly prevented the development of colitis. It reduced the expression of interleukin (IL)-1, IL-6, and tumor necrosis factor (TNF)-α in the colon. Moreover, SH administration alleviated the oxidative stress in the colon of DSS-induced colitis mice, evidenced by the decrease of myeloperoxidase (MPO) activity and malondialdehyde (MDA) level, and increase of ROS level. Furthermore, SH can prevent the decrease ofLactobacillus sp. and population abundance of intestinal flora caused by DSS. CONCLUSION: SH significantly ameliorates the symptoms of DSS-induced colitis mice and the potential mechanism of SH may involve in multiple kinds of metabolic pathway including the regulation of gut microbiota, inflammatory mediators and cytokines.

Suppressive Effect of Huzhentongfeng on Experimental Gouty Arthritis: An In Vivo and In Vitro Study.

BACKGROUND: Huzhentongfeng (HZTF) is an extract from four Chinese medical herbs for treating gout. This study aims to evaluate its antigout activity and preliminary explore its mechanism in vivo and in vitro. METHODS: The rats were intragastrically administered with HZTF for 5 days and then injected 0.1 ml (10 mg) of MSU crystals to their joints for generating a gout model to analyze the paw volume and histopathology of joint synovial tissues of rats with different doses. We also investigated the antioxidant capacity of HZTF in vitro using indication including lipid peroxidation, DPPH·, and ABTS+ radical-scavenging capacity; besides, we used qRT-PCR to measure the effect of HZTF on interleukin (IL)-1ß, caspase-1, NLRP3, and NQO1 expression in hydrogen peroxide-stimulated RAW264.7 macrophages and IL-1ß, IL-6, and tumor necrosis factor (TNF)-α in MSU crystal-induced THP-1 monocytes. Confocal microscopy analysis was used to observe the dimerization of ASC adapter proteins. In addition, we also established quality standard of HZTF by using the high-performance liquid chromatography (HPLC) method. RESULTS: HZTF could significantly suppress the paw swelling and neutrophil infiltration induced by MSU intra-articular injection in rats compared with the control group. HZTF also showed inhibition effects of inflammatory cytokines (IL-1ß, IL-6, and TNF-α) secretion at 25.00 and 50.00 µg/ml in MSU-induced THP-1 cells but showed no effects of IL-1ß, IL-6, and TNF-α mRNA expression in MSU-induced THP-1 cells. Furthermore, confocal microscopy analysis showed that HZTF could prevent the oligomerization of ASC. Moreover, HZTF also showed effects in cell-free and cell-base tests of antioxidant capacity. CONCLUSION: The results prove that HZTF possessed the potential preventive effect against gout arthritis, and the effect may be attributed to its preventing effect on neutrophil infiltration and proinflammatory cytokines secretion such as IL-1ß, IL-6, and TNF-α which were caused by the activation of inflammasome.

A comparison of neural responses in the primary auditory cortex, amygdala, and medial prefrontal cortex of cats during auditory discrimination tasks.

Discriminating biologically relevant sounds is crucial for survival. The neurophysiological mechanisms that mediate this process must register both the reward significance and the physical parameters of acoustic stimuli. Previous experiments have revealed that the primary function of the auditory cortex (AC) is to provide a neural representation of the acoustic parameters of sound stimuli. However, how the brain associates acoustic signals with reward remains unresolved. The amygdala (AMY) and medial prefrontal cortex (mPFC) play keys role in emotion and learning, but it is unknown whether AMY and mPFC neurons are involved in sound discrimination or how the roles of AMY and mPFC neurons differ from those of AC neurons. To examine this, we recorded neural activity in the primary auditory cortex (A1), AMY, and mPFC of cats while they performed a Go/No-go task to discriminate sounds with different temporal patterns. We found that the activity of A1 neurons faithfully coded the temporal patterns of sound stimuli; this activity was not affected by the cats' behavioral choices. The neural representation of stimulus patterns decreased in the AMY, but the neural activity increased when the cats were preparing to discriminate the sound stimuli and waiting for reward. Neural activity in the mPFC did not represent sound patterns, but it showed a clear association with reward and was modulated by the cats' behavioral choices. Our results indicate that the initial auditory representation in A1 is gradually transformed into a stimulus-reward association in the AMY and mPFC to ultimately generate a behavioral choice. NEW & NOTEWORTHY We compared the characteristics of neural activities of primary auditory cortex (A1), amygdala (AMY), and medial prefrontal cortex (mPFC) while cats were performing the same auditory discrimination task. Our results show that there is a gradual transformation of the neural code from a faithful temporal representation of the stimulus in A1, which is insensitive to behavioral choices, to an association with the predictive reward in AMY and mPFC, which, to some extent, is correlated with the animal's behavioral choice.

Diagnostic capacity of electron cyclotron emission imaging system with continuous large observation area on EAST tokamak.

Electron cyclotron emission imaging on EAST provides direct measurements of the 2-D electron temperature dynamics in a continuous large observation area with high temporal and spatial resolution. Besides the normal MHD investigation, a system with a view field large enough to cover the core plasma region has been applied to extract more plasma information, such as the plasma center location, the deposition location of auxiliary heating, and the core toroidal rotation speed. These results solely based on electron cyclotron emission imaging (ECEI) data are consistent with the results of the equilibrium fitting (EFIT), numerical code, and other diagnostics, which indicate the powerful diagnostic capacity of this ECEI system.

Functional mutant GATA4 identification and potential application in preimplantation diagnosis of congenital heart diseases.

Congenital heart diseases (CHDs) affect nearly 1% of all neonates and show an increasing tendency. The complex inheritance patterns and multifactorial etiologies make these defects difficult to be identified before complete manifestation. Genetic screening has identified hundreds of specific mutant sites for CHDs based on cardiac transcriptional factors. GATA4 is a master regulator required for ventral morphogenesis and heart tube formation. Its mutation is most widely studied in CHDs. In the past decades, over 100 GATA4 mutant sites have been reported, but only a few functional sites have been identified. Thus, it is important to distinguish deleterious sites from neutral sites. In silico prediction of functional sites using bioinformatics tools can provide the valuable information, but it is not solid enough. Here, the roles of GATA4 in heart development is discussed in detail and its mutation sites in protein coding region are summarized systematically, providing an integrated resource for GATA4 mutations. Furthermore, we discussed the advantage and disadvantage of different methods for functional mutation identification. Especially, the disease model of induced pluripotent stem cell is emerging as a powerful tool to assess GATA4 mutations in human. In the recent years, single-cell based high-throughput sequencing is being applied in preimplantation diagnosis and assisted reproduction progressively, providing a new strategy for the prevention of congenital diseases as we discussed. Based on functional mutant sites identification, preimplantation diagnosis will contribute to CHDs prevention eventually.

Response properties of neurons in the cat's putamen during auditory discrimination.

The striatum integrates diverse convergent input and plays a critical role in the goal-directed behaviors. To date, the auditory functions of striatum are less studied. Recently, it was demonstrated that auditory cortico-striatal projections influence behavioral performance during a frequency discrimination task. To reveal the functions of striatal neurons in auditory discrimination, we recorded the single-unit spike activities in the putamen (dorsal striatum) of free-moving cats while performing a Go/No-go task to discriminate the sounds with different modulation rates (12.5 Hz vs. 50 Hz) or envelopes (damped vs. ramped). We found that the putamen neurons can be broadly divided into four groups according to their contributions to sound discrimination. First, 40% of neurons showed vigorous responses synchronized to the sound envelope, and could precisely discriminate different sounds. Second, 18% of neurons showed a high preference of ramped to damped sounds, but no preference for modulation rate. They could only discriminate the change of sound envelope. Third, 27% of neurons rapidly adapted to the sound stimuli, had no ability of sound discrimination. Fourth, 15% of neurons discriminated the sounds dependent on the reward-prediction. Comparing to passively listening condition, the activities of putamen neurons were significantly enhanced by the engagement of the auditory tasks, but not modulated by the cat's behavioral choice. The coexistence of multiple types of neurons suggests that the putamen is involved in the transformation from auditory representation to stimulus-reward association.

Behavioral detection of intra-cortical microstimulation in the primary and secondary auditory cortex of cats.

Although neural responses to sound stimuli have been thoroughly investigated in various areas of the auditory cortex, the results electrophysiological recordings cannot establish a causal link between neural activation and brain function. Electrical microstimulation, which can selectively perturb neural activity in specific parts of the nervous system, is an important tool for exploring the organization and function of brain circuitry. To date, the studies describing the behavioral effects of electrical stimulation have largely been conducted in the primary auditory cortex. In this study, to investigate the potential differences in the effects of electrical stimulation on different cortical areas, we measured the behavioral performance of cats in detecting intra-cortical microstimulation (ICMS) delivered in the primary and secondary auditory fields (A1 and A2, respectively). After being trained to perform a Go/No-Go task cued by sounds, we found that cats could also learn to perform the task cued by ICMS; furthermore, the detection of the ICMS was similarly sensitive in A1 and A2. Presenting wideband noise together with ICMS substantially decreased the performance of cats in detecting ICMS in A1 and A2, consistent with a noise masking effect on the sensation elicited by the ICMS. In contrast, presenting ICMS with pure-tones in the spectral receptive field of the electrode-implanted cortical site reduced ICMS detection performance in A1 but not A2. Therefore, activation of A1 and A2 neurons may produce different qualities of sensation. Overall, our study revealed that ICMS-induced neural activity could be easily integrated into an animal's behavioral decision process and had an implication for the development of cortical auditory prosthetics.

Behavioral modulation of neural encoding of click-trains in the primary and nonprimary auditory cortex of cats.

Neural representation of acoustic stimuli in the mammal auditory cortex (AC) has been extensively studied using anesthetized or awake nonbehaving animals. Recently, several studies have shown that active engagement in an auditory behavioral task can substantially change the neuron response properties compared with when animals were passively listening to the same sounds; however, these studies mainly investigated the effect of behavioral state on the primary auditory cortex and the reported effects were inconsistent. Here, we examined the single-unit spike activities in both the primary and nonprimary areas along the dorsal-to-ventral direction of the cat's AC, when the cat was actively discriminating click-trains at different repetition rates and when it was passively listening to the same stimuli. We found that the changes due to task engagement were heterogeneous in the primary AC; some neurons showed significant increases in driven firing rate, others showed decreases. But in the nonprimary AC, task engagement predominantly enhanced the neural responses, resulting in a substantial improvement of the neural discriminability of click-trains. Additionally, our results revealed that neural responses synchronizing to click-trains gradually decreased along the dorsal-to-ventral direction of cat AC, while nonsynchronizing responses remained less changed. The present study provides new insights into the hierarchical organization of AC along the dorsal-to-ventral direction and highlights the importance of using behavioral animals to investigate the later stages of cortical processing.

[Species and ecological control of disease on cultivated Gentiana rigescens in Yunnan].

OBJECTIVE: To find out variety of the fungal diseases of cultivated Gentiana rigescens and provide important basis for prevention. METHODS: The diseases were diagnosed based on field investigate, symptoms observation, pathogen isolation, determination the size of morphological and verification following the Koch's Postulate procedures. RESULTS: Anthracnose (Colletotrichum gloeosporioides), grey mould (Botrytis cinerea), brown spot (Alternaria tenuis), rust (Aecidiumpers), circular spot (Pestalotiopsis), leaf blight (Stemphylium, Ascochyta, Pleospora) and nematodes (Heterodera spp., Meloidogyne spp.) were found on Gentiana rigescens. Anthracnose was the first main disease, the diseased plant rate was over 40% and disease severity was 4 - 5 degree and second disease was rust, incidence of rate was less 10% and other diseases rate was not enough 2%. CONCLUSION: All these diseases on Gentiana rigescens are reported for the first time and Gentiana rigescens is the new host plant of the diseases.

[Correlation and path analysis on agronomic traits of Gentiana rigescens].

OBJECTIVE: The interrelation of yield and agronomic traits of Gentiana rigescens was studied for the germplasm and breeding variety of this species. METHOD: Twelve agronomic traits, root diameter, root length, root number, root biomass, stem diameter, plant height, the first branch number, leaf length, leaf width, leaf length/leaf width ratio, calyx length, and calyx number of G. rigescens from 26 wild populations in Yunnan were determined for correlation analysis, multiple stepwise regression analysis and path analysis. RESULT: Correlation analysis showed that there were significantly positive correlation between the traits of aboveground part and the length, diameter, number, and biomass of the root. Multiple stepwise regression analysis showed that length, width, and number of root, plant height, the first branch number, and the calyx number were the main factors that affected the root biomass. Path analysis showed that the diameter, length, and number of the root, the stem height, and the first branch number had a direct positive effect on the root biomass. CONCLUSION: The traits, such as high and strong stem, high number of first branch number and shrubby shape could be selected for the breeding and high yielding of G. rigescens.

[Seed germination characteristics of Gentiana rigescens].

OBJECTIVE: To investigate the influences of temperature, lightness, storage method, storage time, and gibberellin on seed germination of Gentiana rigescens. METHOD: The germination rates of G. rigescens in different treatments were observed. RESULT AND CONCLUSION: The most suitable temperature for the seed germination was 25 degrees C, at which the germination rate was 76.33%. The effect of lightness on the seeds was significantly; the germination rate of the seed was very low. Under the natural condition, the best storage method was dry storage (within 6 months), which could promote the after-ripening of the seed. 100-1 000 mg x L(-1) gibberellic acid could significantly reduce the seed germination time, and 500 mg x L(-1) gibberellic acid increased the germination rate of the seed to 95.00%.

[Cloning, expression and characterization of gene encoding human stem cell growth factor-alpha and its synergetic effect with rhGM-CSF on proliferation of human umbilical cord mesenchymal stem cells].

To investigate the effect of hSCGF-alpha on human Umbilical Cord Mesenchymal Stem Cells (hUCMSCs), we obtained hSCGF-alpha using genetic engineering, hSCGF-alpha gene was amplified from hUCMSCs cDNA using two-step PCR and was inserted into pET-28a(+) plasmid vector. Induced by IPTG at 20 degrees Celsius for 24 h, the fusion protein expressed in E. coli BL21 (DE3) was mainly existing in soluble form. The recombinant hSCGF-a was purified using NI-NTA affinity chromatography and the purity was up to 90%. The colony forming test revealed that combined use hSCGF-alpha and rmGM-CSF (recombinant murine GM-colony stimulating factor, rmGM-CSF) had granulocyte/macrophage (GM) promoting effects on murine bone marrow GM progenitor. In addition, the results indicated that hSCGF-alpha and rhGM-CSF had stimulatory effect on hUCMSCs and their synergetic effect was the strongest.