Differential responses of rumen and fecal fermentation and microbiota of Liaoning cashmere goats after 2-hydroxy-4-(methylthio) butanoic acid isopropyl ester supplementation.

The 2-hydroxy-4-(methylthio) butanoic acid isopropyl ester (HMBi), a rumen protective methionine, has been extensively studied in dairy cows and beef cattle and has been shown to regulate gastrointestinal microbiota and improve production performance. However, knowledge of the application of HMBi on cashmere goats and the simultaneous study of rumen and hindgut microbiota is still limited. In this study, HMBi supplementation increased the concentration of total serum protein, the production of microbial protein in the rumen and feces, as well as butyrate production in the feces. The results of PCoA and PERMANOVA showed no significant difference between the rumen microbiota, but there was a dramatic difference between the fecal microbiota of the two groups of Cashmere goats after the HMBi supplementation. Specifically, in the rumen, HMBi significantly increased the relative abundance of some fiber-degrading bacteria (such as Fibrobacter) compared with the CON group. In the feces, as well as a similar effect as in the rumen (increasing the relative abundance of some fiber-degrading bacteria, such as Lachnospiraceae FCS020 group and ASV32), HMBi diets also increased the proliferation of butyrate-producing bacteria (including Oscillospiraceae UCG-005 and Christensenellaceae R-7 group). Overall, these results demonstrated that HMBi could regulate the rumen and fecal microbial composition of Liaoning cashmere goats and benefit the host.

MiR-224-5p inhibits osteoblast differentiation and impairs bone formation by targeting Runx2 and Sp7.

Osteoporosis is a complicated multifactorial disorder characterized by low bone mass and deteriorated bone microarchitecture with an elevated fracture risk. MicroRNAs play important roles in osteoblastic differentiation. In the present study, we found that miR-224-5p was markedly downregulated during the osteogenic differentiation of C2C12 cells. Overexpression of miR-224-5p in C2C12 cells inhibited osteoblast activity, as indicated by reduced ALP activity, matrix mineralization and the expression of osteogenic marker genes. Moreover, we demonstrated that Runx2 and Sp7 were direct targets of miR-224-5p. Furthermore, the specific inhibition of miR-224-5p by femoral bone marrow cavity injection with miR-224-5p antagomir prevented ovariectomy-induced bone loss. Finally, we found that the levels of miR-224-5p were markedly elevated in the sera of patients with osteoporosis. Collectively, this study revealed that miR-224-5p negatively regulates osteogenic differentiation by targeting Runx2 and Sp7. It also highlights the potential use of miR-224-5p as a therapeutic target and diagnostic biomarker for osteoporosis. Supplementary information: The online version contains supplementary material available at 10.1007/s10616-023-00593-z.

Natural Product BO-1 as an Inner Responsive Molecule Inhibits Antimicrobial-Resistant Staphylococcus aureus via Synergism.

Multidrug-resistant Staphylococcus aureus, a Gram-positive bacterium that causes several difficult-to-treat human infections, is a considerable threat to global healthcare. We hypothesize that there exist inner responsive molecules (IRMs) which can function synergistically with antibiotics to restore the sensitivity of resistant bacteria to existing antibiotics without inducing new antibiotic resistance. An investigation of the extracts of the Chinese medicinal herb Piper betle L. led to the isolation of six benzoate esters, BO-1-BO-6. Among these, BO-1 as a distinct IRM displayed considerable synergism by potentiating antibacterial activity against five antibiotic-resistant S. aureus strains. Mechanistic studies demonstrated that BO-1 acted as a suppressing drug resistance IRM via inhibiting efflux activity. A combination of BO-1 with ciprofloxacin significantly inhibited resistance to this antibiotic and reversed its resistance in the S. aureus strain. Furthermore, BO-1 effectively enhanced the activity of ciprofloxacin against the efflux fluoroquinolone-resistant S. aureus strain SA1199B that caused infection in two animal models and significantly decreased the inflammatory factors IL-6 and C-reactive protein of the infected mice, thereby showing the practice utility of this approach.

Balance nitrogen and phosphorus efficient removal under carbon limitation in pilot-scale demonstration of a novel anaerobic/aerobic/anoxic process.

Nutrient removal in carbon limited wastewater with high efficiency and energy saving remains a bottleneck for wastewater treatment plants (WWTPs). This study established a pilot-scale anaerobic/aerobic/anoxic (AOA) system with processing capacity of 100 m3/d for the first time. During almost 300 days of stable operation, enhanced nitrogen and phosphorus removal at a C/N of 5 was achieved, and the concentrations of total nitrogen (TN) and total phosphorus (TP) in effluent were 3.60 ± 1.55 and 0.24 ± 0.13 mg/L. Tetrasphaera and Candidatus Competibacter were the dominant phosphorus accumulating organisms (PAOs) and glycogen accumulating organisms (GAOs) in the AOA system. Moreover, the low phosphorus release ensured sufficient intracellular carbon storage by endogenous denitrification, which was the critical factor for nitrogen and phosphorus removal in carbon limited wastewater. The denitrification phosphorus removal (DPR) ability further removed phosphorus and prevented secondary phosphorus release to maintain a low phosphorus concentration in effluent. Finally, rapid start-up, high nutrient removal efficiency and low energy consumption make the proposed AOA process suitable for application in newly constructed and renovated WWTPs.

Adsorption and removal of organophosphorus pesticides from Chinese cabbages and green onions by using metal organic frameworks based on the mussel-inspired adhesive interface.

Based on the mussel-inspired adhesive interface (Fe3O4-g-C3N4@PDA), a novel bionic metal-organic framework (Fe3O4-g-C3N4-PDA@MIL-101) was successfully prepared. The composite featured a high specific surface area and a multi-microchannel structure, as well as strong thermochemical stability. The structural property of Fe3O4-g-C3N4-PDA@MIL-101(Fe) was characterized, and the results indicated that Fe3O4, PDA, and MIL-101(Fe) were uniformly coated on the g-C3N4 surface. The adsorption and desorption of organophosphorus pesticides with Fe3O4-g-C3N4-PDA@MIL-101(Fe) were evaluated by batch experiments. This composite showed high adsorption efficiency and selective removal of coralox, phosalone, and chlorpyrifos. Under the optimal conditions, three organophosphorus pesticides were adsorbed from Chinese cabbage and green onion samples with Fe3O4-g-C3N4-PDA@MIL-101(Fe). The analytical method exhibited high sensitivity (LOD, 0.19-2.34 µg/L; LOQ, 0.65-7.82 µg/L), excellent practicality, and good stability, suggesting that Fe3O4-g-C3N4-PDA@MIL-101 was an ideal candidate magnetic adsorbent for the removal of organophosphorus pesticides in Chinese cabbage and green onion samples.

Mitochondrial-Respiration-Improving Effects of Three Different Gardeniae Fructus Preparations and Their Components.

The processing method for Chinese traditional herbal medicine is "Pao Zhi" in Chinese. This study examined the efficacy of the Pao Zhi on the preparations of Gardeniae Fructus (GF) on a mitochondrial respiratory function in rats. To determine the efficacy of Pao Zhi, we investigated the effects of GF heat processing on mitochondrial respiratory function. To test the GF components, the rats were randomly divided into a geniposide-alone group, crocin-alone group, and combination groups and treated with geniposide and crocin at different ratios. The results showed that a high dose, raw GF was more effective in improving the neurological function, mitochondrial respiratory function, and activities of Na+-K+-ATPase and Ca2+-Mg2+-ATPase than the preparations that underwent heating. Moreover, mitochondrial ROS production was the lowest in the raw GF-treated group. In addition, treatments with crocin and GC3 were more effective than geniposide in improving the functional deficit in MCAO rats. In conclusion, our results suggest that raw GF is the most suitable preparation for the treatment of cerebral ischemia, and its underlying mechanisms may be associated with the improvement of mitochondrial respiratory function, increased activities of Na+-K+-ATPase and Ca2+-Mg2+-ATPase, and reduced oxidative stress in mitochondria. Our findings suggest that raw GF, especially crocin, could be an ideal therapeutic agent for ischemic stroke.

Chinese Medicine Leptochloa chinensis Inhibits the Malignant Behaviors of Renal Cell Carcinoma 786-O Cells by Regulating the mTOR Pathway.

BACKGROUND: Renal cell carcinoma (RCC) is a common malignant tumor of the urinary system that seriously threatens human life and health. This study aims to explore the role of the traditional Chinese medicine Leptochloa chinensis in the pathogenesis of RCC. Meanwhile, this study also revealed the molecular biological mechanism of its antitumor activity. METHODS: Human RCC 786-O cells were cultured in the RPMI-1640 medium, which contains different concentrations of Leptochloa chinensis (1,000, 3,000, and 9,000 µg/ml). MTT and flow cytometry assays were used to detect the viability of 786-O cells. Transwell and wound healing assays were used to detect cell metastasis. The protein expression was observed by western blot analysis. RESULTS: Leptochloa can inhibit cell proliferation and induce apoptosis in RCC 786-O cells. In addition, Leptochloa can weaken the migration and invasion of 786-O cells. More importantly, Leptochloa can block the mTOR pathway by inhibiting the protein expression of p-mTOR. Moreover, the high concentration of Leptochloa chinensis has a better inhibitory effect on 786-O cells. CONCLUSION: The traditional Chinese medicine Leptochloa chinensis inhibits the viability and metastasis of 786-O cells by blocking the mTOR pathway.

Pharmacodynamic effects of the K+ binder patiromer in a novel chronic hyperkalemia model in spontaneously hypertensive rats.

Currently described hyperkalemia (HK) animal models are typically acute and cause significant distress and mortality to the animals, warranting new approaches for studying chronic HK in a more appropriate clinical setting. Using the spontaneously hypertensive rat (SHR) model as a more relevant disease template, as well as surgical (unilateral nephrectomy), dietary (3% potassium [K+ ] supplementation), and pharmacological (amiloride) interventions, we were able to stably induce HK on a chronic basis for up to 12 weeks to serum K+ elevations between 8 and 9 mmol/L, with minimal clinical stress to the animals. Short-term proof-of-concept and long-term chronic studies in hyperkalemic SHRs showed concomitant increases in serum aldosterone, consistent with the previously reported relationship between serum K+ and aldosterone. Treatment with the K+ binder patiromer demonstrated that the disease model was responsive to pharmacological intervention, with significant abrogation in serum K+ , as well as serum aldosterone to levels near baseline, and this was consistent in both short-term and long-term 12-week chronic studies. Our results demonstrate the feasibility of establishing a chronic HK disease state, and this novel HK animal model may be suitable for further evaluating the effects of long-term, K+ -lowering therapies on effects such as renal fibrosis and end-organ damage.

Stimulus-specific hypothalamic encoding of a persistent defensive state.

Persistent neural activity in cortical, hippocampal, and motor networks has been described as mediating working memory for transiently encountered stimuli1,2. Internal emotional states, such as fear, also persist following exposure to an inciting stimulus3, but it is unclear whether slow neural dynamics are involved in this process. Neurons in the dorsomedial and central subdivisions of the ventromedial hypothalamus (VMHdm/c) that express the nuclear receptor protein NR5A1 (also known as SF1) are necessary for defensive responses to predators in mice4-7. Optogenetic activation of these neurons, referred to here as VMHdmSF1 neurons, elicits defensive behaviours that outlast stimulation5,8, which suggests the induction of a persistent internal state of fear or anxiety. Here we show that in response to naturalistic threatening stimuli, VMHdmSF1 neurons in mice exhibit activity that lasts for many tens of seconds. This persistent activity was correlated with, and required for, persistent defensive behaviour in an open-field assay, and depended on neurotransmitter release from VMHdmSF1 neurons. Stimulation and calcium imaging in acute slices showed that there is local excitatory connectivity between VMHdmSF1 neurons. Microendoscopic calcium imaging of VMHdmSF1 neurons revealed that persistent activity at the population level reflects heterogeneous dynamics among individual cells. Unexpectedly, distinct but overlapping VMHdmSF1 subpopulations were persistently activated by different modalities of threatening stimulus. Computational modelling suggests that neither recurrent excitation nor slow-acting neuromodulators alone can account for persistent activity that maintains stimulus identity. Our results show that stimulus-specific slow neural dynamics in the hypothalamus, on a time scale orders of magnitude longer than that of working memory in the cortex9,10, contribute to a persistent emotional state.

The efficacy and safety of electroacupuncture against urinary incontinence after stroke: A protocol for systematic review and meta analysis.

BACKGROUND: Urinary incontinence (UI) is still a persistent challenge in many stroke survivors, affecting the quality of life and emotional being of these individuals. Numerous studies have demonstrated the curative effect of electroacupuncture on post-stroke incontinence, however they were mired with questionable quality and inconsistencies in safety and efficacy. Therefore, the main objective of this meta-analysis is to provide a comprehensive evaluation of the efficacy and safety of electroacupuncture against urinary incontinence after stroke, with a view of providing more reliable evidence-based solutions for UI. METHODS: A systematic literature search will be conducted using PubMed, EMBASE, the Cochrane Central Register of Controlled Trials, Web of Science, and 4 Chinese databases from inception to June 2020 to identify randomized control trials that report on electroacupuncture against urinary incontinence after stroke. Two reviewers will independently identify eligible studies and extract data. The risk of bias of the included randomized control trials will be evaluated according to the Cochrane tool. Risk ratio and 95% confidence intervals will be used to estimate the efficacy of treatment,. and the grading of recommendations, assessment, development, and evaluation approach to rate the certainty of evidence. The statistical heterogeneity will be evaluated by Cochran's Q and the I. Data will be analyzed using Stata software (Version 13.0, Stata Corp, College Station, TX, USA). RESULTS: This study will provide a comprehensive evaluation of the efficacy and safety of electroacupuncture against UI after stroke, with a view of providing more reliable evidence-based solutions for UI. ETHICS AND DISSEMINATION: This work synthesises evidence from previously published studies and does not require ethics review or approval. A manuscript describing the findings will be submitted for publication in a peer-reviewed scientific journal. INPLASY REGISTRATION NUMBER: INPLASY202050073.

[UPLC-Q-TOF/MS for rapid analysis of chemical constituents in Sanhuang tablets].

The qualitative analysis method of ultra performance liquid chromatography tandem quadrupole time of flight mass spectrometry (UPLC-Q-TOF-MS/MS) was established for the chemical constituents in Sanhuang tablets. Waters ACQUITY BEH C18 (2.1 mm×100 mm, 1.7 µm) column was used with 0.1% formic acid solution (A)-0.1% formic acid acetonitrile (B) as the mobile phase for gradient elution. The flow rate was 0.2 mL•min⁻¹; the sample volume was 1 µL and the column temperature was 30 ℃. The high-resolution quadrupole time-flight mass spectrometry was used as detector with electrospray ion source in both positive and negative models, and the dry gas temperature was 325 ℃. Based on the analysis of mass spectrometry and literature reports, 38 compounds were confirmed, including 1 alkaloid, 1 dianthrone compound, 6 tannins, 7 anthraquinone glycosides, 6 anthraquinones and 17 flavonoids. Liquid chromatography-mass spectrometry method is simple, reliable and rapid to identify the chemical compositions of Sanhuang tablets, and it is helpful to reveal its chemical constituents and pharmacodynamic substances.

Glycyrrhizin ameliorates experimental colitis through attenuating interleukin-17-producing T cell responses via regulating antigen-presenting cells.

Glycyrrhizin, a component of Chinese medicine licorice root, has the ability to inhibit the functions of high-mobility group box 1 (HMGB1). While glycyrrhizin is known to have anti-inflammatory activities, the underlying mechanisms by which glycyrrhizin inhibits inflammation during the development of trinitrobenzenesulfonic acid (TNBS)-induced experimental colitis are not well understood. This study systemically examined the regulatory effects of glycyrrhizin on inflammatory response in TNBS-induced murine colitis and explored the potential mechanisms involved in this process. We reported that glycyrrhizin treatment ameliorated colitis and decreased the production of inflammatory mediators HMGB1, IFN-γ, IL-6, TNF-α, and IL-17. In addition, glycyrrhizin regulated responses of dendritic cells (DCs) and macrophages during the development of colitis. Furthermore, administration of glycyrrhizin suppressed the proliferation of Th17 cells in colitis. Moreover, the ability of DCs and macrophages to induce the differentiation of Th17 cells was enhanced in presence of HMGB1, which was inhibited by glycyrrhizin. These results demonstrated that glycyrrhizin alleviated colitis by inhibiting the promotive effect of HMGB1 on DC/macrophage-mediated Th17 proliferation. In conclusion, HMGB1 plays an important role in the development of colitis. As an inhibitor of HMGB1, glycyrrhizin might be a novel therapy for colitis.

Microplastics in Taihu Lake, China.

In comparison with marine environments, the occurrence of microplastics in freshwater environments is less understood. In the present study, we investigated microplastic pollution levels during 2015 in Taihu Lake, the third largest Chinese lake located in one of the most developed areas of China. The abundance of microplastics reached 0.01 × 10(6)-6.8 × 10(6) items/km(2) in plankton net samples, 3.4-25.8 items/L in surface water, 11.0-234.6 items/kg dw in sediments and 0.2-12.5 items/g ww in Asian clams (Corbicula fluminea). The average abundance of microplastics was the highest in plankton net samples from the southeast area of the lake and in the sediments from the northwest area of the lake. The northwest area of the lake was the most heavily contaminated area of the lake, as indicated by chlorophyll-α and total phosphorus. The microplastics were dominated by fiber, 100-1000 µm in size and cellophane in composition. To our best knowledge, the microplastic levels measured in plankton net samples collected from Taihu Lake were the highest found in freshwater lakes worldwide. The ratio of the microplastics in clams to each sediment sample ranged from 38 to 3810 and was negatively correlated to the microplastic level in sediments. In brief, our results strongly suggest that high levels of microplastics occurred not only in water but also in organisms in Taihu Lake.

Functional response properties of VIP-expressing inhibitory neurons in mouse visual and auditory cortex.

Despite accounting for about 20% of all the layer 2/3 inhibitory interneurons, the vasoactive intestinal polypeptide (VIP) expressing neurons remain the least thoroughly studied of the major inhibitory subtypes. In recent studies, VIP neurons have been shown to be activated by a variety of cortico-cortical and neuromodulatory inputs, but their basic sensory response properties remain poorly characterized. We set out to explore the functional properties of layer 2/3 VIP neurons in the primary visual (V1) and primary auditory cortex (A1), using two-photon imaging guided patch recordings. We found that in the V1, VIP neurons were generally broadly tuned, with their sensory response properties resembling those of parvalbumin (PV) expressing neurons. With the exception of response latency, they did not exhibit a significant difference from PV neurons across any of the properties tested, including overlap index, response modulation, orientation selectivity, and direction selectivity. In the A1, on the other hand, VIP neurons had a strong tendency to be intensity selective, which is a property associated with a subset of putative pyramidal cells and virtually absent in PV neurons. VIP neurons had a best intensity that was significantly lower than that of PV and putative pyramidal neurons. Finally, sensory evoked spike responses of VIP neurons were delayed relative to pyramidal and PV neurons in both the V1 and A1. Combined, these results demonstrate that the sensory response properties of VIP neurons do not fit a simple model of being either PV-like broadly tuned or pyramidal-like narrowly tuned. Instead, the selectivity pattern varies with sensory area and can even be, as in the case of low sound intensity responsiveness, distinct from both PV and pyramidal neurons.

New norditerpenoid alkaloids from Aconitum hemsleyanum var. leueanthus.

Four new norditerpenoid alkaloids, leueantines A (1), B (2), C (3), and D (4), were isolated from the roots of Aconitum hemsleyanum var. leueanthus. The structures of 1-4 were established by spectroscopic evidence.

Preclinical evaluations of therapies combining the vascular targeting agent combretastatin A-4 disodium phosphate and conventional anticancer therapies in the treatment of Kaposi's sarcoma.

The antitumor efficacy of the vascular targeting agent combretastatin A-4 disodium phosphate (CA4DP) was evaluated in a xenograft model of Kaposi's sarcoma (KS) grown in athymic mice. Response to CA4DP alone or in combination with localized radiation treatment or systemic chemotherapy (cisplatin or vinblastine) was assessed using a clonogenic cell survival or tumor growth delay assay. Administering increasing doses of CA4DP to tumor-bearing mice resulted in a dose-dependent increase in tumor cell kill. CA4DP also enhanced the antitumor effects of radiation and chemotherapy approximately 10-100-fold. Although single doses of CA4DP as large as 300 mg/kg failed to alter tumor growth, the same total dose, administered as 3 fractions in 5 or 9 days, resulted in significant growth delay. Such repeated CA4DP exposures also significantly increased the response of KS xenografts to cisplatin. These findings suggest that CA4DP ought to be considered as a candidate agent for therapeutic evaluation in AIDS-KS patients.