The lack of PPARα exacerbated the progression of non-alcoholic steatohepatitis in mice with spleen deficiency syndrome by triggering an inflammatory response.

Background: In addition to abnormal liver inflammation, the main symptoms of non-alcoholic steatohepatitis (NASH) are often accompanied by gastrointestinal digestive dysfunction, consistent with the concept of spleen deficiency (SD) in traditional Chinese medicine. As an important metabolic sensor, whether peroxisome proliferator-activated receptor alpha (PPARα) participates in regulating the occurrence and development of NASH with SD (NASH-SD) remains to be explored. Methods: Clinical liver samples were collected for RNA-seq analysis. C57BL/6J mice induced by folium sennae (SE) were used as an SD model. qPCR analysis was conducted to evaluate the inflammation and metabolic levels of mice. PPARα knockout mice (PPARαko) were subjected to SE and methionine-choline-deficient (MCD) diet to establish the NASH-SD model. The phenotype of NASH and the inflammatory indicators were measured using histopathologic analysis and qPCR as well. Results: The abnormal expression of PPARα signaling, coupled with metabolism and inflammation, was found in the results of RNA-seq analysis from clinical samples. SD mice showed a more severe inflammatory response in the liver evidenced by the increases in macrophage biomarkers, inflammatory factors, and fibrotic indicators in the liver. qPCR results also showed differences in PPARα between SD mice and control mice. In PPARαko mice, further evidence was found that the lack of PPARα exacerbated the inflammatory response phenotype as well as the lipid metabolism disorder in NASH-SD mice. Conclusion: The abnormal NR signaling accelerated the vicious cycle between lipotoxicity and inflammatory response in NAFLD with SD. Our results provide new evidence for nuclear receptors as potential therapeutic targets for NAFLD with spleen deficiency.

Exocarpium Citri Grandis ameliorates LPS-induced acute lung injury by suppressing inflammation, NLRP3 inflammasome, and ferroptosis.

ETHNOPHARMACOLOGICAL RELEVANCE: Exocarpium Citri Grandis (ECG), the epicarp of C. grandis 'Tomentosa' which is also known as Hua-Ju-Hong in China, has been widely used for thousands of years to treat inflammatory lung disorders such as asthma, and cough as well as dispelling phlegm. However, its underlying pharmacological mechanisms in acute lung injury (ALI) remain unclear. AIM OF THE STUDY: To explore the therapeutic effect of ECG on ALI and reveal the potential mechanisms based on experimental techniques in vivo and in vitro. MATERIALS AND METHODS: Lipopolysaccharides (LPS) induced ALI in mice and induced RAW 264.7 cell inflammatory model were established to investigate the pharmacodynamics of ECG. ELISA kits, commercial kits, Western Blot, qPCR, Hematoxylin and Eosin (H&E) staining, immunohistochemistry, and immunofluorescence technologies were used to evaluate the pharmacological mechanisms of ECG in ameliorating ALI. RESULTS: ECG significantly attenuated pulmonary edema in LPS-stimulated mice and decreased the levels of IL1ß, IL6, and TNF-α in serum and BALF, reduced MDA and iron concentration as well as increased SOD and GSH levels in lung tissues, and also decreased the ROS level in BALF and Lung tissue. Further pharmacological mechanism studies showed that ECG significantly inhibited mRNA expression of inflammatory signaling factors and chemokines, and down-regulated the expression of TLR4, MyD88, NF-κB p65, NF-κB p-p65 (S536), COX2, iNOS, Txnip, NLRP3, ASC, Caspase-1, JAK1, p-JAK1 (Y1022), JAK2, STAT1, p-STAT1 (S727), STAT3, p-STAT3 (Y705), STAT4, p-STAT4 (Y693), and Keap1, and also up-regulated the expression of Trx-1, Nrf2, HO-1, NQO1, GPX4, PCBP1, and SLC40A1. In the LPS-induced RAW264.7 cell inflammatory model, ECG showed similar results to animal experiments. CONCLUSIONS: Our results showed that ECG alleviated ALI by inhibiting TLR4/MyD88/NF-κB p65 and JAK/STAT signaling pathway-mediated inflammatory response, Txnip/NLRP3 signaling pathway-mediated inflammasome activation, and regulating Nrf2/GPX4 axis-mediated ferroptosis. Our findings provide an experimental basis for the application of ECG.

Effects of stand ages on soil enzyme activities in Chinese fir plantations and natural secondary forests.

Forest type and stand age are important biological factors affecting soil enzyme activities. However, the changes in soil enzyme activities across stand ages and underlying mechanisms under the two forest restoration strate-gies of plantations and natural secondary forests remain elusive. In this study, we investigated the variations of four soil enzyme activities including cello-biohydrolase (CBH), ß-1,4-glucosidase (ßG), acid phosphatase (AP) and ß-1,4-N-acetylglucosaminidase (NAG), which were closely associated with soil carbon, nitrogen, and phosphorus cycling, across Cunninghamia lanceolata plantations and natural secondary forests (5, 8, 21, 27 and 40 years old). The results showed that soil enzyme activities showed different patterns across different forest types. The acti-vities of AP, ßG and CBH in the C. lanceolata plantations were significantly higher than those in the natural secon-dary forests, and there was no significant difference in the NAG activity. In the plantations, AP activity showed a decreasing tendency with the increasing stand ages, with the AP activity in the 5-year-old plantations significantly higher than other stand ages by more than 62.3%. The activities of NAG and CBH decreased first and then increased, and ßG enzyme activity fluctuated with the increasing stand age. In the natural secondary forests, NAG enzyme activity fluctuated with the increasing stand age, with that in the 8-year-old and 27-year-old stand ages being significantly higher than the other stand ages by more than 14.9%. ßG and CBH enzyme activities increased first and then decreased, and no significant difference was observed in the AP activity. Results of the stepwise regression analyses showed that soil predictors explained more than 34% of the variation in the best-fitting models predicting soil enzyme activities in the C. lanceolata plantations and natural secondary forests. In conclusion, there would be a risk of soil fertility degradation C. lanceolata plantations with the increasing stand age, while natural secondary forests were more conducive to maintaining soil fertility.

Observation of plaid-like spin splitting in a noncoplanar antiferromagnet.

Spatial, momentum and energy separation of electronic spins in condensed-matter systems guides the development of new devices in which spin-polarized current is generated and manipulated1-3. Recent attention on a set of previously overlooked symmetry operations in magnetic materials4 leads to the emergence of a new type of spin splitting, enabling giant and momentum-dependent spin polarization of energy bands on selected antiferromagnets5-10. Despite the ever-growing theoretical predictions, the direct spectroscopic proof of such spin splitting is still lacking. Here we provide solid spectroscopic and computational evidence for the existence of such materials. In the noncoplanar antiferromagnet manganese ditelluride (MnTe2), the in-plane components of spin are found to be antisymmetric about the high-symmetry planes of the Brillouin zone, comprising a plaid-like spin texture in the antiferromagnetic (AFM) ground state. Such an unconventional spin pattern, further found to diminish at the high-temperature paramagnetic state, originates from the intrinsic AFM order instead of spin-orbit coupling (SOC). Our finding demonstrates a new type of quadratic spin texture induced by time-reversal breaking, placing AFM spintronics on a firm basis and paving the way for studying exotic quantum phenomena in related materials.

Akkermansia muciniphila supplementation improves glucose tolerance in intestinal Ffar4 knockout mice during the daily light to dark transition.

IMPORTANCE: Alterations in the intestinal environment are associated with various diseases, and FFAR4 is abundantly enriched in the intestine, where it has been shown to have the ability to regulate intestinal hormone secretion and intestinal microbiota; here, we confirmed previous reports. Meanwhile, we found that intestinal FFAR4 regulates glucagon-like peptide 1 secretion by decreasing Akkermansia muciniphila abundance and show that such change is associated with the level of glucose utilization at ZT12 in mice. Intestinal FFAR4 deficiency leads to severely impaired glucose tolerance at the ZT12 moment in mice, and Akkermansia muciniphila supplementation ameliorates the abnormal glucose utilization at the ZT12 moment caused by FFAR4 deficiency, which is very similar to the dawn phenomenon in diabetic patients. Collectively, our data suggest that intestinal Ffar4 deteriorates glucose tolerance at the daily light to dark transition by affecting Akkermansia muciniphila.

Self-assembled silk fibroin cross-linked with genipin supplements microbial carbonate precipitation in building material.

The process of microbially induced carbonate precipitation (MICP) is known to effectively improve engineering properties of building materials and so does silk fibroin (SF). Thus, in this study, an attempt was taken to see the improvement in sand, that is, basic building material coupled with MICP and SF. Urease producing Bacillus megaterium was utilized for MICP in Nutri-Calci medium. To improve the strength of SF itself in bacterial solution, it was cross-linked with genipin at the optimized concentration of 3.12 mg/mL. The Fourier transform infrared (FTIR) spectra confirmed the crosslinking of SF with genipin in bacterial solution. In order to understand how such cross-linking can improve engineering properties, sand moulds of 50 mm3 dimension were prepared that resulted in 35% and 55% more compressive strength than the one prepared with bacterial solution with SF and bacterial solution only, respectively with higher calcite content in former one. The FTIR, SEM, x-ray powder diffraction spectrometry and x-ray photoelectron spectroscopy analyses confirmed higher biomineral precipitation in bacterial solution coupled with genipin cross-linked SF. As the process of MICP is proven to replace cement partially from concrete without negatively influence mechanical properties, SF cross-linked with genipin can provide additional significance in developing low-carbon cement-based composites.

Tetracycline antibiotics: Potential anticancer drugs.

In recent years, research on tetracycline antibiotics has gradually shifted from their antibacterial effects to anticancer effects. Doxycycline, minocycline, and tigecycline as the US Food and Drug Administration (FDA) approved tetracycline antibiotics have been the main subjects of studies. Evidence indicated that they have anticancer properties and are able to control cancer progression through different mechanisms, such as anti-proliferation, anti-metastasis, and promotion of autophagy or apoptosis. In addition, studies have shown that these three tetracycline antibiotics can be utilized in conjunction with chemotherapeutic and targeted drugs to inhibit cancer progression and improve the quality of patient survival. Therefore, doxycycline, minocycline, and tigecycline are taken as examples in this work. Their mechanisms of action in different cancers and related combination therapies are introduced. Their current roles in alleviating the suffering of patients undergoing chemotherapy when used as adjuvant drugs in clinical treatment are also described. Finally, the research gaps and potential research directions at this stage are briefly summarized.

Anti-lipid accumulation effects and mechanism of the extract and isolated compounds from Potentilla longifolia.

Potentilla longifolia is effective in the treatment of hepatitis as a Chinese herb. We firstly evaluated the effect of water extract of P. longifolia (WEPL) on mice with nonalcoholic fatty liver disease (NAFLD) induced by high-fat (HF) diet. The results showed that WEPL reduced HF-induced increases of the serum ALT, AST, TG and TC, and reduced lipid drops of liver tissues to a different extent compared with HF group; WEPL dose-dependently promoted the phosphorylation degrees of AMPK and ACC; WEPL decreased significantly genes expressions of SREBP1α, FAS and SCD1 and increased PPARα and CD36. Then three new (1-3) and 13 known compounds (4-16) were firstly-isolated from the 95% ethanol extract of this plant. Further experiments showed that a new compound (ganyearmcaooside C) showed the best inhibitory effect on lipid accumulation in 3 T3-L1 cells such as reducing the accumulation of oil droplets and triglyceride level, showing new drug potential for related diseases.

LC-MS Metabolite Profiling and the Hypoglycemic Activity of Morus alba L. Extracts.

Morus alba L. is used in traditional Chinese medicine for its anti-diabetic activity; however, the part of the hypoglycemic activity and related active metabolites are still not fully clarified. In this study, the metabolites in the M. alba roots, leaves, twigs, and fruits extracts (70% ethanol extracts) were systematically identified, and their hypoglycemic activity was evaluated by the high-fat diet/streptozotocin-induced 2 diabetes mellitus (T2D) mouse model. A total of 60 high-level compounds, including 16 polyphenols, 43 flavonoids, and one quinic acid, were identified by high-performance liquid chromatography/quadrupole time-of-flight mass spectrometry (HPLC-Q-TOF-MS) combined with the fragmentation pathways of standards and the self-established database. Among them, 23 metabolites were reported for the first time from this plant. In contrast to the extracts of M. alba leaves and fruits, the extracts of roots and twigs displayed significant hypoglycemic activity The glycemia was significantly reduced from 32.08 ± 1.27 to 20.88 ± 1.82 mmol/L and from 33.32 ± 1.98 to 24.74 ± 1.02 mmol/L, respectively, after 4 weeks of treatment with roots and twigs extracts. Compound 46 (morusin), which is a high-level component identified from the extracts of M. alba roots, also displayed significant activity in decreasing the blood glucose level of T2D mice reduced from 31.45 ± 1.23 to 23.45 ± 2.13 mmol/L. In addition, the extracts of roots and twigs displayed significant activity in reducing postprandial glycemia. This work marks the first comparison of the metabolites and hypoglycemic activity of M. alba roots, leaves, twigs, and fruits extracts, and provides a foundation for further development of M. alba extracts as anti-diabetic drugs.

Delavatine A Attenuates OGD/R-Caused PC12 Cell Injury and Apoptosis through Suppressing the MKK7/JNK Signaling Pathway.

Delavatine A (DA) is an unusual isoquinoline alkaloid with a novel skeleton isolated from Chinese folk medicine Incarvillea delavayi. Studies conducted in our lab have demonstrated that DA has potential anti-inflammatory activity in lipopolysaccharide (LPS)-treated BV-2 cells. DA, however, has not been studied for its protective effect on neuronal cells yet. Thus, to explore whether DA can protect neurons, oxygen and glucose deprivation/reperfusion (OGD/R)-injured PC12 cell and middle cerebral artery occlusion/reperfusion (MCAO/R) rat model were used to assess the protective efficacy of DA against OGD/R damaged PC12 cells and MCAO/R injured rats. Our results demonstrated that DA pretreatment (0.31-2.5 µM) dose-dependently increased cell survival and mitochondrial membrane potential (MMP), whereas it lowered the leakage of lactate dehydrogenase (LDH), intracellular cumulation of Ca2+, and overproduction of reactive oxygen species (ROS), and inhibited the apoptosis rate in OGD/R-injured PC12 cells. Western blot demonstrated that DA pretreatment lowered the expression of apoptotic proteins and repressed the activation of the mitogen-activated protein kinase kinase 7 (MKK7)/c-Jun N-terminal kinase (JNK) pathway. It was also found that the neuroprotective efficacy of DA was significantly reversed by co-treatment with the JNK agonist anisomycin, suggesting that DA reduced PC12 cell injury and apoptosis by suppressing the MKK7/JNK pathway. Furthermore, DA oral administration greatly alleviated the neurological dysfunction and reduced the infarct volume of MCAO/R rats. Taken together, DA could ameliorate OGD/R-caused PC12 cell injury and improve brain ischemia/reperfusion (I/R) damage in MCAO/R rats, and its neuroprotection might be attributed to suppressing the MKK7/JNK signaling pathway.

Natural borneol enhances the anti-cerebral ischaemia efficacy of formononetin in MCAO/R rats by promoting its delivery in the brain.

Objectives Due to its high morbidity, high mortality, and high disability, stroke has been the first cause of death and the major cause of adult disability in China. Natural borneol has been widely utilized in Traditional Chinese Medicine to promote drug absorption. Formononetin is a natural isoflavonoid with potent neuroprotective activity but poor brain delivery. Methods This study aimed to screen the optimum proportion that natural borneol promotes formononetin entry into the brain, evaluate the anti-cerebral ischaemia efficacy of formononetin/natural borneol combination in middle cerebral artery occlusion/reperfusion model rats, and clarify the possible mechanism for natural borneol's promoting formononetin delivery in the brain. Key findings Our studies exhibited that natural borneol remarkably promoted formononetin entry into the brain when combined with formononetin in a 1 : 1 molar ratio and notably improved neuro-behavioural scores and reduced the infarct of middle cerebral artery occlusion/reperfusion model rats. This study further discovered that the enhanced anti-cerebral ischaemia effect resulted from natural borneol increasing the permeability of the blood-brain barrier to elevate formononetin concentration in the brain rather than the pharmacodynamic synergy or addition between formononetin and natural borneol. Conclusions The study provides a good strategy to screen drug combinations for the treatment of brain disease by combining natural borneol with other drugs.

Danggui Niantong granules ameliorate rheumatoid arthritis by regulating intestinal flora and promoting mitochondrial apoptosis.

CONTEXT: Danggui Niantong Granules (DGNTG) are a valid and reliable traditional herbal formula, commonly used in clinical practice to treat rheumatoid arthritis (RA). However, the mechanism of its effect on RA remains unclear. OBJECTIVE: An investigation of the therapeutic effects of DGNTG on RA. MATERIALS AND METHODS: Twenty-four male Sprague-Dawley (SD) rats were divided into four groups: control, model, DGNTG (2.16 g/kg, gavage), methotrexate (MTX) (1.35 mg/kg, gavage) for 28 days. The morphology of synovial and ankle tissues was observed by haematoxylin-eosin staining. The responses of mitochondrial apoptosis were assessed by qPCR, Western blotting and immunohistochemical staining. Rat faeces were analysed by 16S rRNA sequencing. RESULTS: Our results showed that DGNTG treatment reduced AI scores (7.83 ± 0.37 vs. 4.67 ± 0.47, p < 0.01) and paw volumes (7.63 ± 0.17 vs. 6.13 ± 0.11, p < 0.01) compared with the model group. DGNTG also increased the expression of Bax (0.34 ± 0.03 vs. 0.73 ± 0.03, p < 0.01), cytochrome c (CYTC) (0.24 ± 0.02 vs. 0.64 ± 0.01, p < 0.01) and cleaved caspase-9 (0.24 ± 0.04 vs. 0.83 ± 0.08, p < 0.01), and decreased bcl-2 (1.70 ± 0.11 vs. 0.60 ± 0.09, p < 0.01) expression. DGNTG treatment regulated the structure of gut microbiota. DISCUSSION AND CONCLUSIONS: DGNTG ameliorated RA by promoting mitochondrial apoptosis, which may be associated with regulating gut microbiota structure. DGNTG can be used as a supplement and alternative drug for the treatment of RA; its ability to prevent RA deserves further study.

Vlasouliodes A-D, four new C30 dimeric sesquiterpenes exhibiting potential inhibition of MCF-7 cells from Vladimiria souliei.

As our ongoing interest to search bioactive dimeric sesquiterpenes from the genus Vladimiria (Asteraceae), the plant of Vladimiria souliei was studied. Based on the repetitive chromatographic fractionation, a chemical investigation on the roots of Vladimiria souliei led to the isolation and the identification of four previously undescribed sesquiterpene dimers, vlasouliodes A-D (1-4). Their chemical structures were elucidated by comprehensive analysis of spectroscopic data, including HRESIMS, 1D and 2D NMR spectroscopic data. The absolute configurations of them were unambiguously established by the experimental and calculated ECD data. In the in vitro biological activity evaluation, 1 and 3 displayed pronounced inhibitory activity against human breast adenocarcinoma cell lines (MCF-7) with IC50 values of 17.12 ± 0.42 µM and 13.12 ± 0.10 µM, respectively. Additionally, treatment with 1 and 3 induced cell apoptosis in MCF-7 cells, down-regulated the expression of Caspase-3 and up-regulated the expression of Cleaved-caspase-3.

Maternal obesity interrupts the coordination of the unfolded protein response and heat shock response in the postnatal developing hypothalamus of male offspring in mice.

Maternal obesity malprograms offspring obesity and associated metabolic disorder. As a common phenomenon in obesity, endoplasmic reticulum (ER) stress also presents early prior to the development. Here, we investigate metabolic effect of early activated hypothalamic ER stress in offspring exposed to maternal obesogenic environment and the underlying mechanism in ICR mice model. We found higher body weight, hyperphagia and defective hypothalamic feeding-circuit in the offspring born to obese dams, with hypothalamic ER stress, and even more comprehensive cell proteotoxic stress were induced during the early postnatal period. However, neonatal inhibition of hypothalamic ER stress worsened the metabolic end. We believe that the uncoordinated interaction between the unfolded protein response and the heat shock response, regulated by heat shock protein 70, might be responsible for the malformed hypothalamic feeding circuit of the offspring exposure to maternal obesogenic conditions and were linked with deleterious metabolism in adulthood, especially when exposure to high-energy conditions.

Cohort profile: the Lanxi Cohort study on obesity and obesity-related non-communicable diseases in China.

PURPOSE: The Lanxi Cohort was established to systematically investigate the aetiology and interplay of body fat distribution and multiple factors with obesity and obesity-related non-communicable diseases in China. PARTICIPANTS: The baseline investigation of the Lanxi Cohort study took place between June 2015 and August 2017 in Lanxi, Zhejiang Province, China. Permanent residents from one urban community and four rural villages were involved in this study. The baseline investigation included questionnaire survey, physical examination, dual-energy X-ray absorptiometry (DXA) scan, blood samples collection and traditional Chinese medicine (TCM) inquiry. FINDINGS TO DATA: A total of 5132 participants, aged 18 to 80 years, were recruited at baseline; among them, 38.7% were men and 64.8% were from the urban area. The mean age was 53.04±12.77 years. The completion rates of physical examination, DXA scan, blood collection and TCM inquiry were 99.9%, 98.5%, 99.9% and 96.5%, respectively. The mean body mass index (BMI) was 23.42±3.20 kg/m2 with 8.1% of the study population being obese (BMI ≥28 kg/m2). The crude prevalence of hypertension, diabetes and metabolic syndrome were 34.9%, 10.0% and 30.4%, respectively. FUTURE PLANS: All participants will be monitored annually for cause-specific mortality and morbidity and hospital admission and will be followed up by in-person survey every 4 years. The baseline population is considered to expand in the future depending on the availability of funding support. ETHICS APPROVAL: This study was approved by the Ethical Committee of the School of Public Health, Zhejiang University.

Antimicrobial activity-guided identification of compounds from the deciduous leaves of Malus doumeri by HPLC-ESI-QTOF-MS/MS.

This paper intends to identify the antimicrobial activity compounds from the deciduous leaves of Malus doumeri (Dong Li Tea) by HPLC-ESI-QTOF-MS/MS. The ethanol extracts of Malus doumeri were partitioned into petroleum ether, dichloromethane, ethyl acetate, n-butanol and water fraction, respectively. The antimicrobial screening experiments showed that ethyl acetate fraction has a certain antibacterial activity by inhibition zone method in vitro. And then we used the HPLC-ESI-QTOF-MS/MS method to verify the identities of bioactive compounds. Finally, 41 compounds were determined and 11 of which were firstly reported in this plant. Notably, compounds (32, 34, 38) are new dihydrochalcones, and three chlorogenic acid analogues (10, 13, 17) may be potential antimicrobial active ingredient. Which is of great significance to the isolation of novel compounds and the discovery of new natural preservative candidates from the deciduous leaves of Malus doumeri.

Association between sleep quality and bone mineral density in Chinese women vary by age and menopausal status.

OBJECTIVE: Sleep quality is closely related to bone health. Aging and estrogen deficiency are known determinants of poor sleep quality and osteoporosis. However, the impact of aging and menopause on the associations between sleep quality and bone mineral density (BMD) remains unclear. This study aimed to examine the association between sleep quality and BMD in Chinese women vary by age groups and menopausal status. METHODS: A total of 2067 women aged 18-80 years were included. Sleep quality was measured using the Pittsburgh Sleep Quality Index (PSQI) and the score >7 was indicative of poor sleep quality. BMD was determined using the dual-energy X-ray absorptiometry. Participants were categorized into three age groups. Multiple linear regression models were conducted to evaluate the associations between sleep quality and BMD. Covariates included in the models were age, menopausal status, weight, height, percent body fat, physical activity, alcohol drinking, calcium supplement use, marital status, education and metabolic diseases. RESULTS: We observed that poor sleep quality was correlated to low total BMD and legs BMD in middle-aged women after adjusting for potential confounders. Furthermore, when we reran the regression models based on menopausal status in middle-aged women, significant associations between BMD and sleep quality were observed in premenopausal and early postmenopausal groups. CONCLUSION: Our findings showed a more robust association between sleep quality and BMD in premenopausal and early menopausal groups. Further studies should be conducted to explore whether sleep quality intervention would improve bone health of women in these periods and prevent osteoporosis in their late life.

Screening and Identification of Antioxidants from Ophiocordyceps xuefengensis (Ascomycetes) by Using DPPH-HPLC-DAD-Q-TOF-MS/MS.

Ophiocordyceps xuefengensis has recently been identified as a sister taxon of O. sinensis and has long been recognized as a folk tonic, food, or invigorant in South China. This study had 3 aims: First, we analyzed the antioxidant activities of 3 kinds of caterpillar fungi; the antioxidative ability of the various caterpillar fungi were in the order of O. xuefengensis > Cordyceps militaris > O. sinensis. Second, we further identified the antioxidant compounds in O. xuefengensis by using various methods: multiwavelength DPPH high-performance liquid chromatography with diode array detection and quadrupole time-of-flight mass spectrometry. In this way, we found and identified 30 compounds that possessed potential antioxidant activity. Among these compounds, 23 compounds were determined to be composed of nucleosides, nucleotides, and amino acids. To our knowledge, several compounds in O. xuefengensis are reported here for the first time. Third, we compared the antioxidant markers in 3 caterpillar fungi. Results showed that the caterpillar fungi share some common antioxidants, while each also had its own characteristic compounds that correspond to its antioxidant capacity. This is to our knowledge the first report of the screening and identification of antioxidants in O. xuefengensis, and these results might provide beneficial contributions to understandings of O. xuefengensis on the basis of its chemical compounds and antioxidant activity.

Anti-fatigue activity of polysaccharides from the fruits of four Tibetan plateau indigenous medicinal plants.

ETHNOPHARMACOLOGICAL RELEVANCE: The fruits of Hippophae rhamnoides L., Lycium barbarum L., Lycium ruthenicum Murr. and Nitraria tangutorum Bobr. are traditional medicinal food of Tibetans and used to alleviate fatigue caused by oxygen deficiency for thousands of years. The present study focused on exploiting natural polysaccharides with remarkable anti-fatigue activity from the four Qinghai-Tibet plateau characteristic berries. MATERIALS AND METHODS: The fruits of Hippophae rhamnoides, Lycium barbarum, Lycium ruthenicum and Nitraria tangutorum were collected from Haixi national municipality of Mongol and Tibetan (N 36.32°, E98.11°; altitude: 3100 m), Qinghai, China. Their polysaccharides (HRWP, LBWP, LRWP and NTWP) were isolated by hot-water extraction, and purified by DEAE-Cellulose ion-exchange chromatography. The total carbohydrate, uronic acid, protein and starch contents of polysaccharides were determined by a spectrophotometric method. The molecular weight distributions of polysaccharides were determined by gel filtration chromatography. Their monosaccharide composition analysis was performed by the method of 1-phenyl-3-methyl-5-pyrazolone (PMP) pre-column derivatization and RP-HPLC analysis. HRWP, LBWP, LRWP and NTWP (50, 100 and 200 mg/kg) were orally administrated to mice once daily for 15 days, respectively. Anti-fatigue activity was assessed using the forced swim test (FST), and serum biochemical parameters were determined by an autoanalyzer and commercially available kits; the body and organs were also weighted. RESULT: LBWP, LRWP and NTWP were mainly composed of glucans and some RG-I pectins, and HRWP was mainly composed of HG-type pectin and some glucans. All the four polysaccharides decreased immobility in the FST, and the effects of LBWP and NTWP were demonstrated in lower doses compared with HRWP and LRWP. There was no significant difference in liver and heart indices between non-treated and polysaccharide-treated mice, but the spleen indices were increased in LBWP and NTWP (200mg/kg) group. Moreover, the FST-induced reduction in glucose (Glc), superoxide dismutase (SOD) and glutathione peroxidase (GPx) and increase in creatine phosphokinase (CK), lactic dehydrogenase (LDH), blood urea nitrogen (BUN), triglyceride (TG) and malondialdehyde (MDA) levels, all indicators of fatigue, were inhibited by HRWP, LBWP, LRWP and NTWP to a certain extent while the effects of LBWP and NTWP were much better than that of HRWP and LRWP at the same dosage. CONCLUSION: Water-soluble polysaccharides HRWP, LBWP, LRWP and NTWP, from the fruits of four Tibetan plateau indigenous berry plants, significantly exhibited anti-fatigue activities for the first time, through triglyceride (TG) (or fat) mobilization during exercise and protecting corpuscular membrane by prevention of lipid oxidation via modifying several enzyme activities. Moreover, it is demonstrated that LBWP and NTWP are more potent than HRWP and LRWP, which were proposed to be applied in functional foods for anti-fatigue and antioxidant potential.

Immunosuppressive activity on the murine immune responses of glycyrol from Glycyrrhiza uralensis via inhibition of calcineurin activity.

CONTEXT: Calcineurin (CN), a unique protein phosphatase, plays an important role in immune regulation. Our laboratory has established an effective molecular drug-screening model based on CN activity. OBJECTIVE: Our aim is to search for an effective immunosuppressant from Glycyrrhiza uralensis (Leguminosae). MATERIALS AND METHODS: As guided by CN inhibitory test, an active compound was purified and identified as glycyrol. Immunosuppressive activity of glycyrol in vitro was assayed by T lymphocytes proliferation and mixed lymphocyte reaction (MLR). In addition, delayed-type hypersensitivity reaction (DTH) and skin allograft test in vivo were also carried out. Further, we have investigated the effect of glycyrol on phorbol 12-myristate 13-acetate (PMA)/ionomycin (Io)-stimulated IL-2 expression in Jurkat cells. RESULTS: The enzymatic assay showed glycyrol (IC(50) = 84.6 µM) inhibited calcineurin activity in a dose-dependent manner. Glycyrol, at the non-cytotoxic concentration, significantly inhibited proliferation of murine spleen T lymphocytes induced by Concanavalin A (Con A) and mixed lymphocyte reaction (MLR) in vitro. In addition, mice treated with glycyrol had shown the dose-dependent decrease in delayed type hypersensitivity (DTH) and prolonged the graft survival by 59% compared to the control group (*p < 0.05). RT-PCR showed glycyrol suppressed IL-2 production in a concentration-dependent manner. DISCUSSION AND CONCLUSION: Our results show the immunosuppressive activity of glycyrol and this activity should be due to its inhibitory effect on CN activity, thereby suppressing IL-2 production and regulating T lymphocytes. Thus, glycyrol could be a candidate for development as a novel immunomodulatory drug.