Angiopterisnodosipetiolata (Marattiaceae), a new fern species from Yunnan, China.

Angiopterisnodosipetiolata Ting Wang tris, H.F.Chen & Y.H.Yan, a new fern of Marattiaceae, is described and illustrated. Morphologically, A.nodosipetiolata is similar to A.chingii with more than one naked pulvinus on the stipe and numerous jointed hairs on the undersides of the mature pinnae. However, the pinnae of A.nodosipetiolata are lanceolate and can reach up to 4-6 pairs, whereas they are elliptic and occur in 2-3 pairs in A.chingii. Phylogenetic and genetic distance analysis, based on the plastid genomes, also indicates that A.nodosipetiolata is not closely related to A.chingii. Currently, there are ca. 500 mature individuals in Gulinqing Nature Reserve and we suggest A.nodosipetiolata should be categorised as an Endangered (EN) species according to the criteria of IUCN.

Amentotaxus×hybridia (Taxaceae), a new natural Amentotaxus hybrid from southeast Yunnan province, China.

During floristic surveys of Taxaceae in Hekou County, Yunnan Province, China, a putative natural hybrid between A.yunnanensis H.L. Li and A.hekouensis L.M. Gao was collected. Morphological and molecular evidence confirms its status as a natural hybrid. Amentotaxus×hybridia L.M. Gao has linear or linear-lanceolate leaves 6-13 cm × 1.0-1.5 cm, white stomatal bands with 34-40 rows on abaxial side, 2.5-3.5 mm, slightly wider than leaf margins; 3-6 seeds borne at the base of the branchlet, peduncle 1.3-1.6 cm long with 3-4 rows of persistent basal bracts.

The Effect of a Three-Level Remote Alliance on Critical Care in Grassroot Areas: A Multi-Center, Retrospective Study.

Purpose: To explore an effective model to promote the homogeneous development of intensive care units (ICUs) in grassroot, impoverished and remote areas. Methods: A three-level remote alliance model (in-place and online assistance) was adopted to guide the cross-talk of ICUs between counties and cities. The observed indicators included the mortality of ICU patients and those with APACHE II scores ≥15 points, deep vein thrombosis, ventilator-associated pneumonia, the completion rate of septic shock goals in 3-hour and 6-hour bundles, and the rates of patient transfers. Results: After the implementation of the remote alliance, there was significant reduction in the mortality of ICU patients in the county and city-level tertiary hospitals (7.6% vs 4.5%, P = 0.004; OR = 1.734, 95% CI 1.189-2.532) and the mortality rates of patients with APACHE II scores ≥15 points (11.9% vs 7.1%, P = 0.004; OR = 1.763, 95% CI 1.189-2.614). There was a significant reduction in the incidence of ventilator-associated pneumonia (0.9% vs 5.0%, P < 0.001) and deep vein thrombosis (52.4% vs 13.6%, P < 0.001). The completion rate of 3-hour bundle therapies for septic shock was significantly improved (95.7% vs 68.4%, P < 0.001), as well as 6-hour bundle therapies for septic shock (97.9% vs 81.6%, P < 0.001). The hospital transfer rate decreased significantly in the grassroots and impoverished areas (2.6% vs 4.7%, P < 0.001). Conclusion: A three-level remote alliance might be helpful in improving the quality of critical care in remote areas and promoting the homogeneous development of disciplines.

Evaluation of the in vitro antioxidant and antitumor activity of extracts from Camellia fascicularis leaves.

Camellia fascicularis is a unique plant rich in bioactive components. However, the isolation of the active substances in C. fascicularis leaves via sequential extraction with solvents of different polarity and the determination of their antioxidant and antitumor activities have not been reported. In this study, the total methanol extract of C. fascicularis leaves was sequentially extracted with different polar solvents, and the corresponding petroleum ether extract (PEE), ethyl acetate extract (EAE), and water extract (WE) were analyzed for their contents in active substances such as flavonoids, polyphenols, polysaccharides, and saponins. The antioxidant ability of the polar extracts was investigated by determining their reducing power and the radical scavenging rate on 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), and hydroxyl radicals, and CCK-8 and Annexin-FITC/propidium iodide staining assays were conducted to investigate their inhibitory effects on HCCLM6 and HGC27 tumor cells. The results showed that PEE had a high saponin content of 197.35 ± 16.21 mg OAE/g, while EAE and WE exhibited a relatively higher polysaccharide content of 254.37 ± 1.99 and 373.27 ± 8.67 mg GE/g, respectively. The EAE demonstrated the greatest reducing power and the strongest clearing abilities on ABTS and DPPH radicals with respective EC50 values of 343.45 ± 20.12 and 14.07 ± 0.06 µg/ml. Moreover, the antitumor ability of the different polar extracts was dose-dependent, with WE showing the most potent inhibitory ability against HCCLM6 and HGC27 cells.

Extraction, characterization, antioxidant and anti-tumor activities of polysaccharides from Camellia fascicularis leaves.

The ultrasonic-assisted extraction of polysaccharides from Camellia fascicularis (PCF) was optimized using response surface methodology. After separation and purification with DEAE-52 cellulose and Sephadex G-200 glucan gel columns, the purified polysaccharide components of PCFa-1 and PCFc-1 were analyzed for their structural characterization, antioxidant and anti-tumor activities in vitro. The results indicated that liquid to material ratio of 42 mL/g, ultrasonic time of 53 min, ultrasonic temperature of 73 °C, and ultrasonic power of 215 W were the optimum extraction conditions for PCF with maximum yields (4.05 %). PCFa-1 and PCFc-1 contained 5.88 % and 9.58 % uronic acid content, with 7.53 and 108.91 kDa of average molecular weights, respectively. The PCFa-1 was mainly constituted of galactose, arabinose, and glucose, while PCFc-1 was primarily composed of arabinose, glucose, galacturonic acid, and rhamnose. Fourier transform infrared spectra revealed that PCFa-1 and PCFc-1 contained typical polysaccharide bands. Scanning electron microscopy showed that the surface of PCFa-1 and PCFc-1 were irregular and clumpy structures. Nuclear magnetic resonance showed that PCFa-1 and PCFc-1 were mainly α-glycosidic bond conformation. Furthermore, the PCFc-1 showed better antioxidant capacities than PCFa-1 against hydroxyl, DPPH, and ABTS radicals and exhibited more potent toxicity on A549 and HepG2 cells. These research results suggested that PCF, especially PCFc-1, possesses great potential as natural antioxidants and anti-tumor drugs.

Harmol promotes α-synuclein degradation and improves motor impairment in Parkinson's models via regulating autophagy-lysosome pathway.

The abnormal accumulation of α-synuclein (α-syn) is a crucial factor for the onset and pathogenesis of Parkinson's disease (PD), and the autophagy-lysosome pathway (ALP) contributes to α-syn turnover. AMP-activated protein kinase (AMPK) and the mammalian target of rapamycin (mTOR) regulate autophagy by initiating the macroautophagy cascade and promoting lysosomal biogenesis via increased transcription factor EB (TFEB) activity. Hence, activation of AMPK-mTOR-TFEB axis-mediated autophagy might promote α-syn clearance in PD. Harmol is a ß-carboline alkaloid that has been extensively studied in a variety of diseases but rarely in PD models. In this study, we aimed to evaluate the effect and underlying mechanism of harmol in PD models in vitro and in vivo. We show that harmol reduces α-syn via ALP in a dose- and time-dependent manner in cell model that overexpressed human A53T mutant α-syn. We also demonstrate that harmol promotes the translocation of TFEB into the nucleus and accompanies the restoration of autophagic flux and lysosomal biogenesis. Importantly, harmol improves motor impairment and down-regulates α-syn levels in the substantia nigra and prefrontal cortex in the α-syn transgenic mice model. Further studies revealed that harmol might activate ALP through AMPK-mTOR-TFEB to promote α-syn clearance. These in vitro and in vivo improvements demonstrate that harmol activates the AMPK-mTOR-TFEB mediated ALP pathway, resulting in reduced α-syn, and suggesting the potential benefit of harmol in the treatment of PD.

Zonation of Landslide Susceptibility in Ruijin, Jiangxi, China.

Landslides are one of the major geohazards threatening human society. The objective of this study was to conduct a landslide hazard susceptibility assessment for Ruijin, Jiangxi, China, and to provide technical support to the local government for implementing disaster reduction and prevention measures. Machine learning approaches, e.g., random forests (RFs) and support vector machines (SVMs) were employed and multiple geo-environmental factors such as land cover, NDVI, landform, rainfall, lithology, and proximity to faults, roads, and rivers, etc., were utilized to achieve our purposes. For categorical factors, three processing approaches were proposed: simple numerical labeling (SNL), weight assignment (WA)-based and frequency ratio (FR)-based. Then 19 geo-environmental factors were respectively converted into raster to constitute three 19-band datasets, i.e., DS1, DS2, and DS3 from three different processes. Then, 155 observed landslides that occurred in the past decades were vectorized, among which 70% were randomly selected to compose a training set (TS1) and the remaining 30% to form a validation set (VS1). A number of non-landslide (no-risk) samples distributed in the whole study area were identified in low slope (<1-3°) zones such as urban areas and croplands, and also added to the TS1 and VS1 in the same ratio. For comparison, we used the FR approach to identify the no-risk samples in both flat and non-flat areas, and merged them into the field-observed landslides to constitute another pair of training and validation sets (TS2 and VS2) using the same ratio of 7:3. The RF algorithm was applied to model the probability of the landslide occurrence using DS1, DS2, and DS3 as predictive variables and TS1 and TS2 for training to obtain the SNL-based, WA-based, and FR-based RF models, respectively. Verified against VS1 and VS2, the three models have similar overall accuracy (OA) and Kappa coefficient (KC), which are 89.61%, 91.47%, and 94.54%, and 0.7926, 0.8299, and 0.8908, respectively. All of them are much better than the three models obtained by SVM algorithm with OA of 81.79%, 82.86%, and 83%, and KC of 0.6337, 0.655, and 0.660. New case verification with the recent 26 landslide events of 2017-2020 revealed that the landslide susceptibility map from WA-based RF modeling was able to properly identify the high and very high susceptibility zones where 23 new landslides had occurred, and performed better than the SNL-based and FR-based RF modeling, though the latter has a slightly higher OA and KC. Hence, we concluded that all three RF models achieve reasonable risk prediction, but WA-based and FR-based RF modeling deserves a recommendation for application elsewhere. The results of this study may serve as reference for the local authorities in prevention and early warning of landslide hazards.

TBN improves motor function and prolongs survival in a TDP-43M337V mouse model of ALS.

Amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD) are serious neurodegenerative diseases. Although their pathogenesis is unclear, the abnormal accumulation of TAR DNA-binding protein of 43 kDa (TDP-43) is a pathological feature that exists in almost all patients. Thus far, there is no drug that can cure ALS/FTLD. Tetramethylpyrazine nitrone (TBN) is a derivative of tetramethylapyrazine, derived from the traditional Chinese medicine Ligusticum chuanxiong, which has been widely proven to have therapeutic effects on models of various neurodegenerative diseases. TBN is currently under clinical investigation for several indications including a Phase II trial of ALS. Here, we explored the therapeutic effect of TBN in an ALS/FTLD mouse model. We injected the TDP-43 M337V virus into the striatum of mice unilaterally and bilaterally, and then administered 30 mg/kg TBN intragastrically to observe changes in behavior and survival rate of mice. The results showed that in mice with unilateral injection of TDP-43M337V into the striatum, TBN improved motor deficits and cognitive impairment in the early stages of disease progression. In mice with bilateral injection of TDP-43M337V into the striatum, TBN not only improved motor function but also prolonged survival rate. Moreover, we show that its therapeutic effect may be through activation of the Akt/mTOR/GSK-3ß and AMPK/PGC-1α/Nrf2 signaling pathways. In summary, TBN is a promising agent for the treatment of ALS/FTLD.

The Tetramethylpyrazine Analogue T-006 Alleviates Cognitive Deficits by Inhibition of Tau Expression and Phosphorylation in Transgenic Mice Modeling Alzheimer's Disease.

T-006, a small-molecule compound derived from tetramethylpyrazine (TMP), has potential for the treatment of neurological diseases. In order to investigate the effect of T-006 prophylactic treatment on an Alzheimer's disease (AD) model and identify the target of T-006, we intragastrically administered T-006 (3 mg/kg) to Alzheimer's disease (AD) transgenic mice (APP/PS1-2xTg and APP/PS1/Tau-3xTg) for 6 and 8 months, respectively. T-006 improved cognitive ability after long-term administration in two AD mouse models and targeted mitochondrial-related protein alpha-F1-ATP synthase (ATP5A). T-006 significantly reduced the expression of phosphorylated-tau, total tau, and APP while increasing the expression of synapse-associated proteins in 3xTg mice. In addition, T-006 modulated the JNK and mTOR-ULK1 pathways to reduce both p-tau and total tau levels. Our data suggested that T-006 mitigated cognitive decline primarily by reducing the p-tau and total tau levels in 3xTg mice, supporting further investigation into its development as a candidate drug for AD treatment.

A new species of Amentotaxus (Taxaceae) from China, Vietnam, and Laos.

A new species Amentotaxus hekouensis L.M. Gao is described as new to science from Hekou, Yunnan of China, Lao Cai of Vietnam and Xiang Khouang of Laos. The new species is similar to A. argotaenia (Hance) Pilg. in linear or linear-lanceolate leaves, stomatal bands white and microsporophylls 6-8, each with 4-6 pollen sacs, but differs from the latter by its larger leaf size with 8-12.5 cm × 0.9-1.4 cm (vs. 2-11 cm × 0.5-1.1 cm in A. argotaenia), long acuminate leaf apex (vs. rounded to sharply triangular in A. argotaenia), stomatal bands with 25-30 rows (vs. 15-25 rows in A. argotaenia), stomatal bands equal to or slightly narrower than marginal bands (vs. narrower than marginal bands in A. argotaenia); pollen-cone racemes borne 1-2 (vs. 2-4 (10) in A. argotaenia), cones in 12-16 pairs (vs. ca. 12 pairs in A. argotaenia). Its distinctive nature has also been confirmed through DNA barcoding analysis of this genus. The new species is provisionally assessed as endangered (EN) due to its restricted distribution, small population size and the prevalence of habitat destruction within its range.

Distribution kinetics of puerarin in rat hippocampus after acute local cerebral ischemia.

Puerarin, isolated from the roots of Pueraria lobata, is widely used for treating cerebral ischemia in China. The time- and dose-dependent distribution characteristics of puerarin in the ischemic hippocampus are unknown. In this study, puerarin concentration was determined by an indirect competitive ELISA using anti-puerarin monoclonal antibody. Area under the curve (AUC0-120 min) of puerarin (80 mg/kg) in the embolic hippocampus was higher than that in the normal hippocampus; the increase was significant only at 40 and 20 mg/kg. The maximum concentration (Cmax) of puerarin in the embolic hippocampus was higher than that in the normal hippocampus at all doses. The increase in both AUC0-120 min and Cmax was dose-dependent. Time to reach the maximum concentration (Tmax) of puerarin in the embolic and normal hippocampus was similar. Although the mean residence time in the embolic hippocampus differed from that in the normal hippocampus at 40 and 80 mg/kg, it was higher in the embolic hippocampus than in the normal hippocampus at 20 mg/kg. This is the first study to report that the time- and dose-dependent distribution characteristics of puerarin in the normal and embolic hippocampus after middle cerebral artery occlusion in rats dictate puerarin dose and duration to treat stroke.

Enzyme-inducing effects of berberine on cytochrome P450 1A2 in vitro and in vivo.

AIMS: Berberine (BER) is an important anti-bacterial drug from Chinese herbal medicine and a novel drug candidate for preclinical development in recent years. Here we provide evidence that the effects of berberine on cytochrome P450 (CYP) 1A2 in vitro and in vivo. MAIN METHODS: Real-time polymerase chain reaction and western blotting analysis were employed to evaluate the CYP1A2 mRNA levels and protein expression. The enzyme activity was assessed by the metabolic rate of phenacetin to acetaminophen by LC-MS/MS method. KEY FINDINGS: The results indicated that the CYP1A2 mRNA expression and enzyme activity in HepG2 cells after treated with BER (4.5µg/ml) exhibited a significant induction (16.11-fold and 5.0-fold, respectively), which was consistent with those on rat liver microsomes (4.5-fold and 1.98-fold, respectively) by BER induction (10mg/kg/day, i.p.) ex vivo. Beside, BER induced CYP1A2 activity with increases in AUC0-t and Cmax of acetaminophen and the Ke and t1/2 of phenacetin after oral administration of phenacetin (p<0.05) in vivo. SIGNIFICANCE: This study firstly reported the induction effect of BER on rats CYP1A2 by intraperitoneal route. But, BER didn't show significant induction effect on CYP1A2 by high-dose orally administrating to rats for 6 consecutive days due to the extremely low bioavailability. The potential drug-drug interactions were supposed to happen when the liver exposed to high dose of BER in vivo by changing administration route.

Pharmacokinetics of geniposide by monoclonal antibody-based icELISA in mice after oral administration of Huanglian-Jiedu-Tang.

Geniposide, Geniposide, the main active component in extracts of Gardenia jasminoides ELLIS., is one of the main components of Huanglian-Jiedu-Tang (HJT). This study aimed to validate an indirect competitive enzyme-linked immunosorbent assay (icELISA) based on monoclonal antibodies (mAb) against geniposide (anti-geniposide mAb), which was developed by our lab, and apply the assay to study the pharmacokinetics of geniposide in HJT in mice. Blood samples were drawn from mice at predetermined time points after oral administration of HJT in three dosages. A linear correlation was obtained for geniposide concentrations in the range from 1.17 to 37.50 µg/mL. The intra-day and inter-day precision values of the icELISA method were well within the recommended range (≤10%). The recovery rates ranged from 99.74 to 102.40%. Stability studies showed that geniposide sample solutions were intact for 12 h. The Tmax and mean residence time (MRT) of geniposide of the three groups were consistent with previous data. The results suggest that a reliable and effective method was established and could be applied to the study of the pharmacokinetics of geniposide in HJT.

Development of an enzyme-linked immunosorbent assay based on anti-puerarin monoclonal antibody and its applications.

An enzyme-linked immunosorbent assay (ELISA) was developed, and its application in immunoaffinity column chromatography was studied using a monoclonal antibody (MAb) against puerarin. Splenocytes isolated from a female BALB/c mouse immunised with a puerarin-bovine serum albumin (BSA) conjugate were fused with SP2/0 myeloma cells. The hybridoma cell line secreting MAb against puerarin (AA9) was acquired by screening and limiting dilution. The antibody generated was highly specific for puerarin with <0.01% cross-reactivity with over 50 structurally related chemicals, except for baicalein (51.8%). Using AA9, we developed an immunoassay for puerarin with a linear detection range of 10ng/ml to 1µg/ml. This assay system was further validated using intra- and inter-assays and recovery experiments. In addition, puerarin levels in both formulated Chinese medicines and biological samples were determined with high sensitivity and efficiency. Finally, we developed and validated protocols for knocking puerarin out of its parent medicine completely. In conclusion, we successfully developed a reliable ELISA and an immunoaffinity column for puerarin detection and knockout, which are useful tools for exploring the role of puerarin in formulated Chinese medicines.