The therapeutic effect of wasp venom (Vespa magnifica, Smith) and its effective part on rheumatoid arthritis fibroblast-like synoviocytes through modulating inflammation, redox homeostasis and ferroptosis.

ETHNOPHARMACOLOGICAL RELEVANCE: Rheumatoid arthritis (RA) is a chronic inflammatory disease that is related to the aberrant proliferation of fibroblast-like synoviocytes (FLS). Wasp venom (WV, Vespa magnifica, Smith), an insect secretion, has been used to treat RA in Chinese Jingpo national minority's ancient prescription. However, the potential mechanisms haven't been clarified. AIM OF THE STUDY: The purposes of this paper were two-fold. First, to investigate which was the best anti-RA effective part of WV-I (molecular weight less than 3 kDa), WV-II (molecular weight 3-10 kDa) and WV-III (molecular weight more than 10 kDa) that were separated from WV. Second, to explore the underlying molecular mechanism of WV and WV-II that was best effective part in RA. MATERIALS AND METHODS: The wasps were electrically stimulated and the secretions were collected. WV-I, WV-II and WV-III were acquired by ultracentrifuge method according to molecular weight. Next, WV, WV-I, WV-II and WV-III were identified by HPLC. Functional annotation and pathway analysis of WV used to bioinformatics analysis. RNA-seq analyses were constructed to identify differentially expressed genes (DEGs). GO and KEGG pathway analyses were performed by Metascape database. STRING was used to analyze the PPI network from DEGs. Next, PPI network was visualized using Cytoscape that based on MCODE. The pivotal genes of PPI network and MCODE analysis were verified by qRT-PCR. Subsequently, MH7A cells were performed by MTT assay to evaluate the ability of inhibiting cell proliferation. Luciferase activity assay was conducted in HepG2/STAT1 or HepG2/STAT3 cells to assess STAT1/3 sensitivity of WV, WV-I, WV-II and WV-III. Additionally, interleukin (IL)-1ß and IL-6 expression levels were detected by ELISA kits. Intracellular thioredoxin reductase (TrxR) enzyme was evaluated by TrxR activity assay kit. ROS levels, lipid ROS levels and Mitochondrial membrane potential (MMP) were assessed by fluorescence probe. Cell apoptosis and MMP were measured by using flow cytometry. Furthermore, the key proteins of JAK/STAT signaling pathway, protein levels of TrxR and glutathione peroxidase 4 axis (GPX4) were examined by Western blotting assay. RESULTS: RNA-sequencing analysis of WV displayed be related to oxidation-reduction, inflammation and apoptosis. The data displayed that WV, WV-II and WV-III inhibited significantly cells proliferation in human MH7A cell line compared to WV-I treatment group, but WV-III had no significant suppressive effect on luciferase activity of STAT3 compared with IL-6-induced group. Combined with earlier reports that WV-III contained major allergens, we selected WV and WV-II further to study the mechanism of anti-RA. In addition, WV and WV-II decreased the level of IL-1ß and IL-6 in TNF-α-induced MH7A cells via inactivating of JAK/STAT signaling pathway. On the other hand, WV and WV-II down-regulated the TrxR activity to produce ROS and induce cell apoptosis. Furthermore, WV and WV-II could accumulate lipid ROS to induce GPX4-mediated ferroptosis. CONCLUSIONS: Taken together, the experimental results revealed that WV and WV-II were potential therapeutic agents for RA through modulating JAK/STAT signaling pathways, redox homeostasis and ferroptosis in MH7A cells. Of note, WV-II was an effective part and the predominant active monomer in WV-II will be further explored in the future.

Examine sex differences in autism spectrum disorder in school-aged children and adolescents with fluent language.

There are noteworthy sex disparities in the prevalence of autism spectrum disorders (ASD), while findings regarding the sex differences in core symptoms are inconsistent. There are few relevant studies on sex differences in mainland China. This study was dedicated to a deeper understanding of the impact of sex differences on the clinical presentation of ASD with fluent language. We retrospectively studied 301 children with ASD (58 females) and utilized raw scores from the ADI-R and ADOS and the intelligence quotient (IQ) to measure symptomatology. Based on the Full-Scale IQ (FS-IQ), a binary split of average, above-average IQ (high-IQ), and below-average IQ (low IQ) occurs at 85. Across the entire sample, males and females are comparable in the FS-IQ, while males scored higher in the Perceptual Reasoning Index (PRI) (F = 7.812, p = 0.006). ADI-R did not find any statistically significant sex differences in the diagnostic cutoff score satisfaction or the raw domain scores. While a significant effect of sex on ADOS social affect domain scores was found in the total sample [λ = 0.970, partial η2 = 0.030, F (3,295) = 3.019, p = 0.030]. Tests of between-subjects effects revealed that males scored higher than females mainly in the ADOS reciprocal social interaction subcategory (partial η2 = 0.022, F = 6.563, p = 0.011). Stratified analysis revealed that the effect of sex on ADOS reciprocal social interaction subcategory scores only significant in the low-IQ children with ASD (partial η2 = 0.092, F = 10.088, p = 0.002). In general, overall cognitive functioning is similar across males and females with ASD, while males have a higher perceptual reasoning ability. Females with ASD are more likely to have comorbid intellectual impairment than males, and they could require additional intervention support. Autistic children with low IQs are more likely to exhibit sex differences in their core symptoms than children with high IQs. Intelligence plays a key role in sex-based differences in the core symptoms of ASD.

A multimodal machine learning system in early screening for toddlers with autism spectrum disorders based on the response to name.

Background: Reduced or absence of the response to name (RTN) has been widely reported as an early specific indicator for autism spectrum disorder (ASD), while few studies have quantified the RTN of toddlers with ASD in an automatic way. The present study aims to apply a multimodal machine learning system (MMLS) in early screening for toddlers with ASD based on the RTN. Methods: A total of 125 toddlers were recruited, including ASD (n = 61), developmental delay (DD, n = 31), and typical developmental (TD, n = 33). Procedures of RTN were, respectively, performed by the evaluator and caregiver. Behavioral data were collected by eight-definition tripod-mounted cameras and coded by the MMLS. Response score, response time, and response duration time were accurately calculated to evaluate RTN. Results: Total accuracy of RTN scores rated by computers was 0.92. In both evaluator and caregiver procedures, toddlers with ASD had significant differences in response score, response time, and response duration time, compared to toddlers with DD and TD (all P-values < 0.05). The area under the curve (AUC) was 0.81 for the computer-rated results, and the AUC was 0.91 for the human-rated results. The accuracy in the identification of ASD based on the computer- and human-rated results was, respectively, 74.8 and 82.9%. There was a significant difference between the AUC of the human-rated results and computer-rated results (Z = 2.71, P-value = 0.007). Conclusion: The multimodal machine learning system can accurately quantify behaviors in RTN procedures and may effectively distinguish toddlers with ASD from the non-ASD group. This novel system may provide a low-cost approach to early screening and identifying toddlers with ASD. However, machine learning is not as accurate as a human observer, and the detection of a single symptom like RTN is not sufficient enough to detect ASD.

Chemical Constituents from Chloranthus anhuiensis and Their Cytotoxic Activities.

Three hiherto unknown phenylpropanoid compounds, namely (7S,8R)-1-(1-ethoxy-2-hydroxypropyl)-2-methoxy-3,4-(methylenedioxy)benzene (1), (7S,8S)-1-(1-ethoxy-2-hydroxypropyl)-2-methoxy-3,4-(methylenedioxy)benzene (2), and (7S,8R)-1-(1-methoxy-2-hydroxypropyl)-2-methoxy-3,4-(methylenedioxy)benzene (3), along with 12 known compounds (4 - 15) were obtained from the extract of whole plant of Chloranthus anhuiensis. Among them, 7 and 13 were obtained from nature for the first time. The structures of these natural compounds were characterized by extensive spectroscopic analysis and calculated electronic circular dichroism (ECD) data. Furthermore, their cytotoxic and neuroprotective activities were evaluated using MDA-MB-231, 4T1, HepG2, and PC12 cell lines. Compounds 8 and 13 exhibited moderate cytotoxic activities against MDA-MB-231 cell line with the IC50 values of 39.7 and 25.8 µm, respectively. And all the isolated compounds have no neuroprotective activities.

Sesquiterpenoids from Chloranthus anhuiensis with Neuroprotective Effects in PC12 Cells.

Glutamate-induced excitotoxicity plays a vital role in neurodegenerative diseases. Neuroprotection against excitotoxicity has been considered as an effective experimental approach for preventing and/or treating excitotoxicity-mediated diseases. In the present study, six new sesquiterpenoids (1-6) and 26 known compounds of this type (7-32) were isolated and characterized from the whole plants of Chloranthus anhuiensis. Chlorantolide A (1) is the first example of a 5,6- seco-germacrane-type sesquiterpenoid, while phacadinane E (2) is a rare 4,5- seco-cadinane-type sesquiterpenoid. The structures of the new compounds were determined by spectroscopic analysis and by calculations of electronic circular dichroism (ECD) spectra. Their neuroprotective effects in mediating glutamate-induced PC12 cell apoptosis were evaluated. Compound 26 exhibited potent neuroprotective activity with an EC50 value of 3.3 ± 0.9 µM. Using Hoechst 33258 staining, a caspase-3 activity assay, and Western blot analysis it was demonstrated that this compound reduces the apoptosis of PC12 cells through inhibition of caspase-3 activity, while activating the Akt signaling pathway.

[Effects of Bone Marrow Mesenchymal Stem Cells on the Biological Characteristics of Migrating and Homing in Multiple Myeloma Cells].

OBJECTIVE: To construct a co-culture system for bone marrow mesenchymal stem cells (BMMSC) and multiple myeloma (MM) cells, and to investigate the effects of co-cultured BMMSC on the migrating and homing of multiple myeloma cells. METHODS: The BMMSC from the transgenic mice with green fluorescent protein (GFP) fetal bone were cultured by adherent screening. A co-culture system of BMMSC and MM cell line XG-7 cells was constracted, the proliferation and apoptosis of cells were determined by trypan blue exclusion and Annexin V/PI, respectively, MDC staining was employed to detect the autophagy. The moving direction distribution of molecule in BMMSC and XG-7 cells labeled with PE-CD138 in co-culture process were observed dinamically by confocal microscopy. RESULTS: After co-culture with GFP-BMMSC, the resistance of XG-7 cells to apoptosis and autophagy were enhanced; at the same time, their proliferation increased. Apoptosis rates of XG-7 cells directly and indirectly co-cultured with BMMSC were (6.23 ± 0.12)% and (6.97 ± 0.03)% respectively, which were lower than that of XG-7 cells cultured alone (17.90 ± 1.46)% (P < 0. 01). There was low level of autophagy in XG-7 cells co-cultured with BMMSC. XG-7 cells are highly polarized and contained a specialized membrane domain with specific protein and lipid components to contact with BMMSC under confocal microscope. After methyl-ß-cyclodextrin treatment, the molecules were normally enriched in the specialized domain. CONCLUSION: BMMSC can protect XG-7 cells from apoptosis and autophagy, and obviously promote the proliferation of XG-7 cells, and can influence the migrating and homing of multiple myeloma cells.

[Quantifying direct N2O emissions from paddy fields during rice growing season in China: model and input data validation].

The models on direct N2O emissions from rice paddies under different water regimes developed by the authors were validated against field measurements in China reported in 2005-2007 and in other regions. In flooding rice paddies (F), N2O emission predicted by the model was consistent with previous reports in other regions. Under the water regime of flooding-midseason drainage-reflooding (F-D-F), the model developed in this study was comparable to that established by using worldwide database. The models also well fitted N2O emissions from rice paddies under the water regime of flooding-midseason drainage-reflooding-moisture but without waterlogging (F-D-F-M) in China. Consistency of rice production data derived from the database of this study with those reported in previous studies suggests that the model input data of rice production had high reliability. The input data showed that water management and nitrogen input regimes have greatly changed in rice paddies since the 1950s. During the 1950s-1970s, about 20%-25% of the rice paddy was continuous water logging, and 75%-80% under the water regime of F-D-F. Since the 1980s, about 12%-16%, 77% and 7%-12% of paddy fields were under the water regimes of F, F-D-F and F-D-F-M, respectively. Total N input during the rice growing season averaged 87.49 kg x hm(-2) in the 1950s and 224.64 kg x hm(-2) in the 1990s. Chemical N input during the rice growing season has increased from 37.4 kg x hm(-2) in the 1950s to 198.8 kg x hm(-2) in the 1990s, accounting for 43% and 88% of the seasonal total N inputs, respectively. Manure N input was applied at stable rate, ranging from 45.2 kg x hm(-2) to 48.2 kg x hm(-2) during the 1950s-1970s, but thereafter it decreased over time. The contribution of manure N to total N inputs has decreased from 52% in the 1950s to 9% in the 1990s. Crop residue N retained during the rice growing season has increased from 4.9 kg x hm(-2) in the 1950s to 6.3 kg x hm(-2) in the 1980s. A high spatial variation of nitrogen inputs during the rice growing season was found in the 1950s-1970s, while it was not pronounced in the 1980s-1990s. Overall, the results of this study suggest that the models could be used to quantify direct N2O emissions from rice paddies under various water regimes in China.

[Quantifying direct N2O emissions from paddy fields during rice growing season in China: model application].

Based on statistical analysis of field N2O measurements in rice paddies in China, the models on direct N2O emissions under different water regimes were established. After successes in model validation and input data verification, the models were used to quantify changes in direct N2O emissions from paddy fields during the rice growing season in mainland China between the 1950s and the 1990s. Due to increases in rice planting area and nitrogen input and changes in water regime, the models predicted that seasonal N2O-N emissions from rice paddies have increased from 9.55 Gg each year in the 1950s to 32.26 Gg N2O-N in the 1990s, which was accompanied by the increase in rice yield over the period 1950s-1990s. During the period 1950s-1990s, seasonal N2O-N emissions from rice paddies have increased, on average, at a rate of 6.74 Gg per decade. Seasonal N2O fluxes in rice paddies were estimated to be 0.32 kg x hm(-2) in the 1950s and 1.00 kg x hm(-2) in the 1990s, which accounted for 0.37% and 0.46% of the seasonal total N inputs, respectively. The uncertainties in N2O estimate were estimated to be 59.8% in the 1950s and 37.5% in the 1990s. Seasonal N2O emissions from rice paddies in the region of middle and lower Yangtze River contributed 51% -56% to its national total. In the 1990s, N2O emissions during the rice growing season accounted for 8%-11% of the reported annual total of N2O emissions from croplands in China, suggesting that paddy rice development could have contributed to mitigating agricultural N2O emissions in the past decades. However, seasonal N2O emissions would be increased given that saving-water irrigation and nitrogen inputs are increasingly adopted in rice paddies in China.

Cytotoxic cardenolides from Streptocaulon griffithii.

A new cardenolide, 3-O-(beta-glucopyranosyl)acovenosigenin A (1), was isolated from the roots of Streptocaulon griffithii, together with eight known cardenolides, compounds 2-9. All compounds showed significant in vitro inhibition of the proliferation of the human gastrointestinal cancer cell line HGC-27, with IC50 values in the range of ca. 20-564 nM, taxol being used as positive control. Compound 1 was also found to be strongly cytotoxic against other human tumor cell lines, including A549, MCF-7, and Hela cells.