The de novo missense mutation F224S in GABRB2, identified in epileptic encephalopathy and developmental delay, impairs GABAAR function.

Genetic variants in genes encoding subunits of the γ-aminobutyric acid-A receptor (GABAAR) have been found to cause neurodevelopmental disorders and epileptic encephalopathy. In a patient with epilepsy and developmental delay, a de novo heterozygous missense mutation c.671T>C (p.F224S) was discovered in the GABRB2 gene, which encodes the ß2 subunit of GABAAR. Based on previous studies on GABRB2 variants, this new GABRB2 variant (F224S) would be pathogenic. To confirm and investigate the effects of this GABRB2 mutation on GABAAR channel function, we conducted transient expression experiments using GABAAR subunits in HEK293T cells. The GABAARs containing mutant ß2 (F224S) subunit showed poor trafficking to the cell membrane, while the expression and distribution of the normal α1 and γ2 subunits were unaffected. Furthermore, the peak current amplitude of the GABAAR containing the ß2 (F224S) subunit was significantly smaller compared to the wild type GABAAR. We propose that GABRB2 variant F224S is pathogenic and GABAARs containing this ß2 mutant reduce response to GABA under physiological conditions, which could potentially disrupt the excitation/inhibition balance in the brain, leading to epilepsy.

Changes in microbial composition explain the contrasting responses of glucose and lignin decomposition to soil acidification in an alpine grassland.

Soil acidification often suppresses microbial growth and activities, resulting in a negative impact on soil organic carbon (C) decomposition. While the detrimental effects of acidification on soil and plant properties have been extensively studied, less attention has been paid on the shifts in soil microbial communities and their influences of the decomposition of organic C with different chemical complexities. Taking advantage of an acid addition experiment in a Tibetan alpine meadow, here we examined the response of soil microbial communities to soil acidification and microbial effect on the decomposition of organic C with different chemical complexities (i.e., glucose and lignin, representing labile and recalcitrant C respectively). We found that soil acidification had no impact on microbial respiration and microbial abundance even though it decreased bacterial diversity significantly. Soil acidification increased the relative abundance of some microbial taxa, like Alphaproteobacteria and Acidobacteriia in bacteria increased by 36 %, 284 %, and Eurotiomycetes, Sordariomycetes and Leotiomycetes in fungi increased by 145 %, 279 % and 12.7-fold, but decreased the relative abundance of Acidimicrobiia by 33 % in highest acid addition treatment. Changes in microbial communities (bacterial and fungal community composition, the diversity of bacterial community and the ratio of fungi to bacteria) are significantly related to the decomposition of glucose and lignin. More specifically, soil acidification decreased the decomposition of glucose but increased the decomposition of lignin, indicating a trade-off between the decomposition of labile and recalcitrant soil organic C under soil acidification. Overall, shifts in microbial communities under soil acidification might be accompanied by an increased ability to break down more recalcitrant C. This trade-off between the decomposition of labile and recalcitrant C may change soil C quality under future acid deposition scenarios.

Periplaneta americana extract attenuates hepatic fibrosis progression by inhibiting collagen synthesis and regulating the TGF-ß1/Smad signaling pathway.

INTRODUCTION: Liver fibrosis is the damage repair response following chronic liver diseases. Activated hepatic stellate cells (HSCs) are the main extracellular matrix (ECM)-producing cells and key regulators in liver fibrosis. Periplaneta americana shows prominent antifibrotic effects in liver fibrosis; however, the underlying mechanisms remain undetermined. This study aimed to elucidate the therapeutic effects of P. americana extract (PA-B) on liver fibrosis based on the regulation of the TGF-ß1/Smad signal pathway. MATERIAL AND METHODS: HSCs and Sprague Dawley rats were treated with TGF-ß1 and CCl4, respectively, to establish the hepatic fibrosis model in vitro and in vivo. The effect of PA-B on liver rat fibrosis was evaluated by biochemical (serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), hyaluronic acid (HA), laminin (LN), collagen type IV (Col-IV), pro-collagen type III (PC-III)) and histological examinations. Further, fibrogenic markers expression of alpha smooth muscle actin (α-SMA), collagen type I (Col-I), and collagen type III (Col-III), and the TGF-ß1/Smad pathway-related factors were assessed by immunofluorescence (IF), real time quantitative polymerase chain reaction (RT-qPCR), and western blotting (WB). RESULTS: Treatment of HSC-T6 cells with PA-B suppressed the expression of α-SMA, Col-I, and Col-III, downregulated the expression of TGF-ß1 receptors I and II (TßR I and TßR II, respectively), Smad2, and Smad3, and upregulated Smad7 expression. PA-B mitigates pathologic changes in the rat model of liver fibrosis, thus alleviating liver index, and improving liver function and fibrosis indices. The effects of PA-B on the expression of α-SMA, Col-I, Col-III, TßR I, TßR II, Smad2, Smad3, and Smad7 were consistent with the in vitro results, including reduced TGF-ß1 expression. CONCLUSIONS: The therapeutic effect of PA-B on liver fibrosis might involve suppression of the secretion and expression of TGF-ß1, regulation of the TGF-ß1/Smad signaling pathway, and inhibition of collagen production and secretion.

Molecular Cloning, Expression Analysis, and Functional Analysis of Nine IbSWEETs in Ipomoea batatas (L.) Lam.

Sugar Will Eventually be Exported Transporter (SWEET) genes play an important regulatory role in plants' growth and development, stress response, and sugar metabolism, but there are few reports on the role of SWEET proteins in sweet potato. In this study, nine IbSWEET genes were obtained via PCR amplification from the cDNA of sweet potato. Phylogenetic analysis showed that nine IbSWEETs separately belong to four clades (Clade I~IV) and contain two MtN3/saliva domains or PQ-loop superfamily and six~seven transmembrane domains. Protein interaction prediction showed that seven SWEETs interact with other proteins, and SWEETs interact with each other (SWEET1 and SWEET12; SWEET2 and SWEET17) to form heterodimers. qRT-PCR analysis showed that IbSWEETs were tissue-specific, and IbSWEET1b was highly expressed during root growth and development. In addition to high expression in leaves, IbSWEET15 was also highly expressed during root expansion, and IbSWEET7, 10a, 10b, and 12 showed higher expression in the leaves. The expression of SWEETs showed a significant positive/negative correlation with the content of soluble sugar and starch in storage roots. Under abiotic stress treatment, IbSWEET7 showed a strong response to PEG treatment, while IbSWEET10a, 10b, and 12 responded significantly to 4 °C treatment and, also, at 1 h after ABA, to NaCl treatment. A yeast mutant complementation assay showed that IbSWEET7 had fructose, mannose, and glucose transport activity; IbSWEET15 had glucose transport activity and weaker sucrose transport activity; and all nine IbSWEETs could transport 2-deoxyglucose. These results provide a basis for further elucidating the functions of SWEET genes and promoting molecular breeding in sweet potato.

Silencing of CPNE1-TRAF2 Axis Restrains the Development of Pancreatic Cancer.

BACKGROUND: Copine 1 (CPNE1) acts as a promoter in the progression of many kinds of cancers with the exception of pancreatic cancer (PC). This research is designed to probe the function of the CPNE1-tumor necrosis factor receptor-associated factor 2 (TRAF2) axis in PC. METHODS: In vivo and in vitro models of PC were constructed, and a series of biological function tests, including MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide], colony formation, flow cytometry, and immunohistochemistry, were performed. RESULTS: The level of CPNE1 elevated dramatically in PC cells. Downregulation of CPNE1in PC cells resulted in the inhibition of colony formation and proliferation. In addition, the silencing of CPNE1 induced the G1/S arrest and apoptosis in PC cells. Additionally, TRAF2 positively interacted with CPNE1 in PANC cells. CPNE1 silencing also inhibited the growth of tumors in in vivo mouse models. Functional experiments revealed that the anti-tumor effect of CPNE1 silencing was counteracted by TRAF2 overexpression, and the tumor-promoting effect of TRAF2 overexpression was reversed by CPNE1 silencing. CONCLUSIONS: In summary, our findings indicate that the silencing of the CPNE1-TRAF2 axis restrains PC development.

Dual RNA-Seq Analysis Pinpoints a Balanced Regulation between Symbiosis and Immunity in Medicago truncatula-Sinorhizobium meliloti Symbiotic Nodules.

Legume-rhizobial symbiosis initiates the formation of root nodules, within which rhizobia reside and differentiate into bacteroids to convert nitrogen into ammonium, facilitating plant growth. This process raises a fundamental question: how is plant immunity modulated within nodules when exposed to a substantial number of foreign bacteria? In Medicago truncatula, a mutation in the NAD1 (Nodules with Activated Defense 1) gene exclusively results in the formation of necrotic nodules combined with activated immunity, underscoring the critical role of NAD1 in suppressing immunity within nodules. In this study, we employed a dual RNA-seq transcriptomic technology to comprehensively analyze gene expression from both hosts and symbionts in the nad1-1 mutant nodules at different developmental stages (6 dpi and 10 dpi). We identified 89 differentially expressed genes (DEGs) related to symbiotic nitrogen fixation and 89 DEGs from M. truncatula associated with immunity in the nad1-1 nodules. Concurrently, we identified 27 rhizobial DEGs in the fix and nif genes of Sinorhizobium meliloti. Furthermore, we identified 56 DEGs from S. meliloti that are related to stress responses to ROS and NO. Our analyses of nitrogen fixation-defective plant nad1-1 mutants with overactivated defenses suggest that the host employs plant immunity to regulate the substantial bacterial colonization in nodules. These findings shed light on the role of NAD1 in inhibiting the plant's immune response to maintain numerous rhizobial endosymbiosis in nodules.

Molecular and Clinicopathological Characteristics of Lung Cancer Concomitant Chronic Obstructive Pulmonary Disease (COPD).

Introduction: Chronic obstructive pulmonary disease (COPD) and lung cancer often coexist, but its pathophysiology and genomics features are still unclear. Methods: In this study, we retrospectively collected lung cancer concomitant COPD (COPD-LC) and non-COPD lung cancer (non-COPD-LC) patients, who performed next generation sequencing (NGS) and had clinicopathological information simultaneously. The COPD-LC data from the TCGA cohort were collected to conduct further analysis. Results: A total of 51 COPD-LC patients and 88 non-COPD-LC patients were included in the study. Clinicopathological analysis showed that proportion of male gender, older age, and smoking patients were all substantially higher in COPD-LC group than in non-COPD-LC group (all P<0.01). Comparing the genomic data of the two groups in our cohort, COPD-LC had higher mutation frequency of LRP1B (43% vs 9%, P = 0.001), EPHA5 (24% vs 1%, P = 0.002), PRKDC (14% vs 1%, P = 0.039), PREX2 (14% vs 0%, P = 0.012), and FAT1 (14% vs 0%, P = 0.012), which had a relationship with improved tumor immunity. Immunotherapy biomarker of PD-L1 positive expression (62.5% vs 52.0%, P = 0.397) and tumor mutation burden (TMB, median TMB: 7.09 vs 2.94, P = 0.004) also were higher in COPD-LC. In addition, RNA data from TCGA further indicated tumor immunity increased in COPD-LC. Whereas, COPD-LC had lower frequency of EGFR mutation (19% vs 50%, P = 0.013) and EGFR mutant COPD-LC treated with EGFR-TKI had worse progression-free survival (PFS) (HR = 3.52, 95% CI: 1.27-9.80, P = 0.01). Conclusion: In this retrospective study, we first explored molecular features of COPD-LC in a Chinese population. Although COPD-LC had lower EGFR mutant frequency and worse PFS with target treatment, high PD-L1 expression and TMB indicated these patients may benefit from immunotherapy.

Identification of anti-hepatic fibrosis components in Periplaneta americana based on spectrum-effect relationship and chemical component separation.

Periplaneta americana (PA) is used as a traditional medicine for hepatic diseases such as hepatic fibrosis in China. However, the relationship between the corresponding therapeutic effect and the chemical composition is still unclear. In this study, spectrum-effect relationship and chemical component separation were used to discover the potential of anti-hepatic fibrosis components of PA. The fingerprints of 10 batches of samples were established using HPLC, and the anti-hepatic fibrosis effect was determined using HSC-T6 cells. The spectrum-effect relationship between common peaks and efficacy values was established using partial least squares analysis. Partial peaks in the fingerprints were identified, including X4 (9,12-heptadecanedenoic acid glyceride), X5 (nonadecanoic acid methyl ester), X6 (glyceryl oleate), X7 (13,16,19-eicosatrienoic acid), X9 (linoleic acid), X10 (9,12,15-octadecatrienoic acid glyceride), X12 (hexadecanoic acid), X13 (oleic acid), and X14 (octadecanoic acid), and their anti-hepatic fibrosis activity was tested to verify the results of spectrum-effect relationships. The results showed that X4 , X6 , X7 , and X10 were the active ingredients of PA. This work successfully identified the partial anti-hepatic fibrosis components of PA, which can be used to explain the material basis for the PA anti-hepatic fibrosis effect.

Influences of climate change on area variation of Qinghai Lake on Qinghai-Tibetan Plateau since 1980s.

Qinghai-Tibetan Plateau is the most sensitive region to global warming on Earth. Qinghai Lake, the largest lake on the plateau, has experienced evident area variation during the past several decades. To quantify the area changes of Qinghai Lake, a satellite-based survey based on Landsat images from the 1980s to 2010s has been performed. In addition, meteorological data from all the seven available stations on Qinghai-Tibetan Plateau has been analyzed. Area of Qinghai Lake shrank ~2% during 1987-2005, and then increased ~3% from 2005-2016. Meanwhile, the average annual temperature increased 0.319 °C/10 y in the past 50 years, where the value is 0.415 °C/10 y from 2005-2016. The structural equation modeling (SEM) shows that precipitation is the primary factor influencing the area of Qinghai Lake. Moreover, temperature might be tightly correlated with precipitation, snow line, and evaporation, thereby indirectly causes alternations of the lake area. This study elucidated the significant area variation of water body on the Qinghai-Tibetan Plateau under global warming since 1980s.

Biochar increases plant growth and alters microbial communities via regulating the moisture and temperature of green roof substrates.

Green roofs have increasingly been designed and applied to relieve environmental problems, such as water loss, air pollution as well as heat island effect. Substrate and vegetation are important components of green roofs providing ecosystem services and benefiting the urban development. Biochar made from sewage sludge could be potentially used as the substrate amendment for green roofs, however, the effects of biochar on substrate quality and plant performance in green roofs are still unclear. We evaluated the effects of adding sludge biochar (0, 5, 10, 15 and 20%, v/v) to natural soil planted with three types of plant species (ryegrass, Sedum lineare and cucumber) on soil properties, plant growth and microbial communities in both green roof and ground ecosystems. Our results showed that sludge biochar addition significantly increased substrate moisture, adjusted substrate temperature, altered microbial community structure and increased plant growth. The application rate of 10-15% sludge biochar on the green roof exerted the most significant effects on both microbial and plant biomass by 63.9-89.6% and 54.0-54.2% respectively. Path analysis showed that biochar addition had a strong effect on microbial biomass via changing the soil air-filled porosity, soil moisture and temperature, and promoted plant growth through the positive effects on microbial biomass. These results suggest that the applications of biochar at an appropriate rate can significantly alter plant growth and microbial community structure, and increase the ecological benefits of green roofs via exerting effects on the moisture, temperature and nutrients of roof substrates.

Application of insect-proof nets in pesticide-free rice creates an altered microclimate and differential agronomic performance.

BACKGROUND: Insect-proof nets are commonly used in crop production and scientific research because of their environmental, economic, and agronomic benefits. However, insect-proof nets can unintentionally alter the microclimate inside the screenhouse and therefore greatly affect plant growth and yield. To examine the microclimate and agronomic performance of pesticide-free rice under insect-proof nets, two-year field experiments were carried out in 2011 and 2012. METHODS: In the present study, the experiment was conducted by using a split-plot design considering the cultivation environment (open field cultivation (OFC) and insect-proof nets cultivation (IPNC)) as the main plot and the varieties as the subplot (Suxiangjing3 and Nanjing44). RESULTS: IPNC significantly reduced the air speed and solar radiation, and slightly increased the daytime soil temperature, daytime air temperature, and nighttime relative humidity. By contrast, the nighttime soil temperature, nighttime air temperature, and daytime relative humidity were relatively unaffected. The grain yield of both rice cultivars decreased significantly under IPNC, which was largely attributed to the reduced panicle number. The reduced panicle number was largely associated with the decreased maximum tiller number, which was positively correlated with the tillering rate, time of tillering onset, and tillering cessation for both rice cultivars under IPNC. In addition, dry matter accumulation significantly decreased for both rice cultivars under IPNC, which was mainly caused by the decreased leaf area duration resulting from the reduced leaf area index. By contrast, the mean net assimilation rate was relatively unaffected by IPNC. DISCUSSION: Insect-proof nets altered the microclimate in comparison with OFC by reducing the air speed and changing the radiation regime, which significantly affected dry matter production and yield of both japonica rice cultivars. Our results indicated that cultivation measures that could increase the tillering rate and the maximum tiller number under IPNC would lead to a significant increase in panicle number, ultimately increasing grain yield. In addition, maintaining a high leaf area duration by increasing the leaf area index would be important to compensate for the dry matter accumulation losses under IPNC. These findings are critical to provide a theoretical basis for improving agronomic performance of pesticide-free rice under IPNC.

Development of hypoxia control device for cell culture in vitro / 口腔疾病防治

Objective@#To simulate the hypoxic environment of cells in vivo more approximately, a hypoxia control device which can control oxygen concentration automatically for cell culture in vitro was developed. @* Methods@#In vitro cell cultur e hypoxia control device consisted of hardware and software which were controlled by computer. The hardware was composed of oxygen detection and control system, nitrogen gas pipeline, cell culture and anti-jamming device, and the software was based on Clanguage. When oxygen controlle rreads the oxygen concent ration more than the set value, the nitrogen gas pipeline opens and nitrogen enter into the incubator; when reaching the set value, the nitrogen gas pipeline closes and nitrogen is not allowed to pass through. Circularly, a constant oxygen concentration was reached and maintained. This device was further verified by comparing the set value of the equipment with figures of the oxygen meter and by monitoring fluctuation condition after the oxygen concentration reaching the set value. @* Results @# There was no significant difference between the oxygen concentration set in the in vitro cell culture hypoxia control device and the actual oxygen concentration in the measured box (P > 0.05), and the accuracy of the device to set the value of the simulation up to ± 0.5% of the requirements to meet the system accuracy requirements, and can remain stable for a long time. @*Conclusion @#The device can effectively control the oxygen concentration to the required requirements, to meet the conditions in the hypoxic conditions for in vitro cell culture.

[Physiological and ecological effects of sulfur fertilization on soybean].

A pot experiment with 3 levels of elemental sulfur (0, 30, and 60 mg S x kg(-1)) showed that sulfur fertilization on soybean increased the side roots number by 8.6% - 33.2%, root dry weight by 6.6% - 34.3%, root nodules number by 2.7% - 35.9% and dry weight by 13.0% - 75.7%, chlorophyll content by 0.4 - 3.9 unit, and yield per plant by 7.3% - 12.8%. Sulfur fertilization also increased the amount of soil bacteria, fungi and actinomycetes and the activities of peroxidase, urease, neutral phosphatase and polyphenoloxidase significantly. The effects of sulfur supply differed with its application rate, and 30 mg S x kg(-1) was more appropriate for getting high soybean yield.

[Effects of elemental sulphur and dicyandiamide on mitigating NO3- -N leaching loss from vegetable soil].

In a pot experiment with allium as test plant and NH4HCO3 as nitrogen source, this paper studied the effects of element sulphur (S0) and dicyandiamide (DCD) on mitigating the NO3- -N leaching loss from soil and on soil inorganic nitrogen (NO3- -N and NH4+ -N) content. The results showed that within the 12 weeks of the experiment, the cumulative leaching loss of soil NO3- -N in treatments S0 + DCD and S0 was 83%-86% and 83% lower, while that of soil NH4+ -N was 16.8-21.0 mg x pot(-1) and 20.4-25.0 mg x pot(-1) higher than CK, respectively, and the cumulative loss of soil (NH4+ + NO3-)-N was 60% lower. By the end of the experiment, soil inorganic nitrogen content in treatments S0 + DCD and S0 was 79.9%-85.4% and 74.9-82.6% higher than CK, respectively. The cumulative leaching loss of inorganic N in treatment S0 + DCD was 4.6%-14.4% and 15.4%-30.1% lower, and the soil inorganic nitrogen content by the end of the experiment was 6.1% and 16.8%-36.0% higher than that of treatments S0 and DCD, respectively. Similar results were obtained when S0 was replaced by Na2S2O3, but not by Na2SO4. The fact that the application of S0 could obviously decrease the NO3- -N leaching loss from soil could be contributed to the inhibitory effects of S2O3(2-) and S4O6(2-) originated from S0 oxidation in soil on the nitrification of NH4+ -N. S0 could retard the decomposition of DCD due to the effect of its oxidized products S2O3(2-) and S4O6(2-), and thus, extend the inhibitory effect of DCD on NH4+ -N nitrification. It is suggested that S0 combined with DCD could be used as an effective nitrification inhibitor to control the NO3- -N leaching loss from vegetable soils.

[Dynamics of dry deposition velocities (Vd) of atmospheric SO2 on rapeseed/rice rotation systems in selected area of south China].

With the profile meteorological data collected from the farmland micro-meteorological experimental substation, Experiment Station of Red Earth Ecology, Yingtan, Jiangxi, Chinese Academy of Sciences during rapeseed/rice rotation from Nov. 1998 to Oct. 1999, the surface layer turbulence characteristic parameters (u*, theta*, L) and the atmospheric SO2 dry deposition velocities (Vd) were calculated. The results indicated that the hourly mean Vd of SO2 dry deposition was ranged from 0.124 to 0.897 cm.s-1 (mean +/- SE = 0.507 +/- 0.167 cm.s-1), and its regular dynamic pattern was as follows: it was always higher at daytime than at night, and the mean Vd of SO2 dry deposition during March-August(0.611 cm.s-1) was obviously greater than that during Sept.-Dec. and Jan.-Feb. (0.401 cm.s-1). The mean Vd of SO2 dry deposition during various crop growth stages was decreased as the order of rice (0.605 +/- 0.093 cm.s-1) > rapeseed (0.491 +/- 0.166 cm.s-1) > follow (0.342 +/- 0.174 cm.s-1).