Role of TRPC6 in apoptosis of skeletal muscle ischemia/reperfusion injury.

BACKGROUND: Skeletal muscle ischaemia-reperfusion injury (IRI) is a prevalent condition encountered in clinical practice, characterised by muscular dystrophy. Owing to limited treatment options and poor prognosis, it can lead to movement impairments, tissue damage, and disability. This study aimed to determine and verify the influence of transient receptor potential canonical 6 (TRPC6) on skeletal muscle IRI, and to explore the role of TRPC6 in the occurrence of skeletal muscle IRI and the signal transduction pathways activated by TRPC6 to provide novel insights for the treatment and intervention of skeletal muscle IRI. METHODS: In vivo ischaemia/reperfusion (I/R) and in vitro hypoxia/reoxygenation (H/R) models were established, and data were comprehensively analysed at histopathological, cellular, and molecular levels, along with the evaluation of the exercise capacity in mice. RESULTS: By comparing TRPC6 knockout mice with wild-type mice, we found that TRPC6 knockout of TRPC6 could reduced skeletal muscle injury after I/R or H/R, of skeletal muscle, so as therebyto restoringe some exercise capacity inof mice. TRPC6 knockdown can reduced Ca2+ overload in cells, therebyo reducinge apoptosis. In additionAdditionally, we also found that TRPC6 functionsis not only a key ion channel involved in skeletal muscle I/R injury, but also can affects Ca2+ levels and then phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin (PI3K/Akt/mTOR) signalling pathway. by knocking downTherefore, knockdown of TRPC6, so as to alleviated the injury inducedcaused by skeletal muscle I/R or and H/R. CONCLUSIONS: These findingsdata indicate that the presence of TRPC6 exacerbatescan aggravate the injury of skeletal muscle injury after I/Rischemia/reperfusion, leading towhich not only causes Ca2+ overload and apoptosis., Additionally, it impairsbut also reduces the self- repair ability of cells by inhibiting the expression of the PI3K/Akt/mTOR signalling pathway. ETo exploringe the function and role of TRPC6 in skeletal muscle maycan presentprovide a novelew approachidea for the treatment of skeletal muscle ischemia/reperfusion injury.

Membrane-associated RING-CH 7 inhibits stem-like capacities of bladder cancer cells by interacting with nucleotide-binding oligomerization domain containing 1.

BACKGROUND: Cancer stem-like capacities are major factors contributing to unfavorable prognosis. However, the associated molecular mechanisms underlying cancer stem-like cells (CSCs) maintain remain unclear. This study aimed to investigate the role of the ubiquitin E3 ligase membrane-associated RING-CH 7 (MARCH7) in bladder cancer cell CSCs. METHODS: Male BALB/c nude mice aged 4-5 weeks were utilized to generate bladder xenograft model. The expression levels of MARCHs were checked in online databases and our collected bladder tumors by quantitative real-time PCR (q-PCR) and immunohistochemistry (IHC). Next, we evaluated the stem-like capacities of bladder cancer cells with knockdown or overexpression of MARCH7 by assessing their spheroid-forming ability and spheroid size. Additionally, we conducted proliferation, colony formation, and transwell assays to validate the effects of MARCH7 on bladder cancer CSCs. The detailed molecular mechanism of MARCH7/NOD1 was validated by immunoprecipitation, dual luciferase, and in vitro ubiquitination assays. Co-immunoprecipitation experiments revealed that nucleotide-binding oligomerization domain-containing 1 (NOD1) is a substrate of MARCH7. RESULTS: We found that MARCH7 interacts with NOD1, leading to the ubiquitin-proteasome degradation of NOD1. Furthermore, our data suggest that NOD1 significantly enhances stem-like capacities such as proliferation and invasion abilities. The overexpressed MARCH7 counteracts the effects of NOD1 on bladder cancer CSCs in both in vivo and in vitro models. CONCLUSION: Our findings indicate that MARCH7 functions as a tumor suppressor and inhibits the stem-like capacities of bladder tumor cells by promoting the ubiquitin-proteasome degradation of NOD1. Targeting the MARCH7/NOD1 pathway could be a promising therapeutic strategy for bladder cancer patients.

Short-term postmortem interval estimation by detection of apoptosis-related protein in skin.

Time-of-death extrapolation has always been one of the most important issues in forensic practice. For a complicated case in which a corpse is destroyed with little evidence, judging the time of death of the deceased is a major challenge, which also enables criminals to escape legal sanctions. To find a method to roughly judge the time of death of a corpse with only a small amount of skin tissue, in this study, we established an early death model by using mice; furthermore, the postmortem interval was estimated by determining the protein and mRNA levels of Bax and Bcl-2 in the skin. In this process, 0 h after death was used as the control group, and the expression levels of Bax and Caspase-3 reached the maximum value at 8-12 h, while Bcl-2, as an inhibitor of apoptosis protein, peaked after 24 h. The mRNA expression levels of related proteins in postmortem skin tissues were also different. The results of these data indicate that the protein and mRNA levels of Bax and Bcl-2 in the skin have potential application in early time-of-death estimation.

Enhancing Electrochemical Performance of Aluminum-Oxygen Batteries with Graphene Aerogel Cathode.

Aluminum-oxygen batteries (AOBs) own the benefits of high energy density (8.14 kWh kg-1), low cost, and high safety. However, the design of a cathode with high surface area, structure integrity, and good catalytic performance is still challenging for rechargeable AOBs. Herein, the fabrication of a robust self-supporting cathode using 3D graphene aerogel (3DGA) for rechargeable AOBs is demonstrated. Electroanalysis showed that the 3DGA presented good catalytic activity in both oxygen reduction and evolution reactions, which allowed the AOB to operate for >90 cycles with low overpotentials at a current density of 0.2 mA cm-2, and a high Coulombic efficiency of ca. 99% using ionic liquid as electrolyte. In comparison, the cell with the carbon paper cathode can only cycle for 50 rounds. The excellent cyclic performance can be attributed to the porous structure, large surface area, good electric conductivity, and catalytic activity of the 3DGA, which is prospective to be applied for other metal-air batteries, fuel cells, and supercapacitors.

Study on the mechanism of nitrapyrin microcapsule suspension effectively improving nitrification inhibition rate in black soil.

Nitrification inhibitors (NIs) have been widely applied to inhibit nitrification and reduce N2O emissions in agriculture. However, there are still some shortcomings, e.g. short effective periods, large applying amounts, low effectiveness, easy deactivation and different effect. Thus, a nitrapyrin microcapsule suspension (CPCS) was used as a new experimental material to elaborate its effects on nitrogen transformation and microbial response mechanisms in black soil by cultivation experiments with six treatments of no fertilization (CK), urea, urea+ 0.2 % CPES, urea+ 0.1 % CPCS, urea+ 0.2 % CPCS, and urea+ 0.3 % CPCS. The content of ammonium, nitrate nitrogen, functional microbial activity, degradation rate and adsorption characteristics of CPCS in the soil at different incubating times were determine. Compared with the nitrapyrin emulsifiable concentrate (CPEC) treatment, the degradation rate of CPCS decreased by 21.54 %, the half-life increased by 10.2 days, and the adsorption rate of nitrapyrin on black soil decreased more than 6-fold. CPCS effectively inhibited the transformation of ammonium nitrogen to nitrate nitrogen within more than 42 days. CPCS had a negative effect on amoA gene abundance and a positive effect on nrfA gene abundance. The research results provide a basic theoretical support for the application of CPCS on black soil.

Second generation androgen receptor antagonist, TQB3720 abrogates prostate cancer growth via AR/GPX4 axis activated ferroptosis.

Purpose: Prostate cancer (PCa) poses a great threat to humans. The study aimed to evaluate the potential of TQB3720 in promoting ferroptosis to suppress prostate cancer, providing a theoretical basis for PCa therapy. Methods: PCa cells and nude mice models were divided into TQB3720, enzalutamide (ENZ), and control groups. Sulforhodamine B assay, colony formation assessment, organoids culture system, and the CCK8 assay were used for detecting proliferation. Western blot assay was processed to detect the expression of androgen receptor (AR), ferroptosis, and apoptosis-related genes. Flow cytometry was applied to measure the intracellular ROS levels. ELISA was performed to determine the cellular oxidized glutathione (GSSG) and malondialdehyde (MDA) levels. RT-qPCR was conducted to detect the mRNA expression of genes in AR signaling. BODIPYTM™ 581/591 was processed for detection of intracellular lipid peroxidation levels. The interaction of AR with other translational factor complex proteins was explored using Co-immunoprecipitation (Co-IP), and the chromatin immunoprecipitation (ChIP) assay was performed to detect the binding of AR-involved translational complex to downstream genes promoter. Luciferase reporter assay was conducted to examine the translation activity of GPX4 promoter, and immunohistochemistry (IHC) was conducted to analyze the levels of c-MYC, Ki-67 and AR in TQB3720-treated cancer tissues. Results: Here, we found TQB3720 inhibits the growth of prostate cancer in vitro and in vivo. TQB3720 treatment induced intracellular levels of GSSG and MDA significantly, by which hints AR antagonist caused ferroptosis-related cell death. Moreover, molecular evidence shown TQB3720 regulates downstream of AR signaling by binding AR resulting in inhibition of AR entry into the nucleus. Additional, we also proved that TQB3720 abrogates the interaction between AR and SP1 and leads to decrease GPX4 transcription. Conclusion: TQB3720 promotes ferroptosis in prostate cancer cells by reducing the AR/SP1 transcriptional complex binding to GPX4 promoter. As a result, it is suggested to be a potential drug for clinic prostate cancer treatment.

A study on and adsorption mechanism of ammonium nitrogen by modified corn straw biochar.

Using corn stover as raw material, the adsorption mechanism of ammonium nitrogen by biochar prepared by different modification methods was studied. The biochar was characterized by Fourier transform infrared spectroscopy, surface-area analysis and scanning electron microscopy. The results showed that the adsorption of NH 4 + - N by different modified biochars confirmed the quasi-second-order kinetic equation (R 2 > 0.95, p ≤ 0.05), the adsorption isotherms of the Langmuir equation (R 2 ≥ 0.96, p ≤ 0.05). ΔG θ < 0, ΔH θ > 0 indicated that the adsorption of NH 4 + - N by different modified biochars was a spontaneous endothermic reaction. With the increase in adsorption temperature, the adsorption capacity of biochar to ammonium nitrogen increased gradually. The adsorption was monolayer adsorption and was controlled by a fast reaction. Both KOH and FeCl3 modified biochars significantly improved the adsorption capacity of NH 4 + - N , and the adsorption mechanism was different. The adsorption capacity of NH 4 + - N by FeCl3 modified biochars mainly increased the specific surface area and micropore volume. The adsorption of ammonium nitrogen after KOH modification primarily depended on the wealthy oxygen-containing functional groups. The adsorption effect of ammonium nitrogen modified by KOH was better than that of biochar modified by FeCl3.

OTUCD: Unsupervised GCN based metagenomics non-overlapping community detection.

Metagenomics is a discipline that studies the genetic material of all tiny organisms in the biological environment. In recent years, the interaction between metagenomic microbial communities, the transfer of horizontal genes, and the dynamic changes of microbial ecosystems have attracted more and more attention. It is of great significance to use the community detection algorithm to divide the metagenomic microbes into modules, and it has a positive guiding role for the follow-up research on human, drug, microbial interaction study and drug prediction and development. At present, there are challenges in mining the effective information hidden in large-scale microbial sequence data. The non-linear characteristics and non-scalability of microbial sequence data still bother people. This paper proposes an end-to-end unsupervised GCN learning model OTUCD (Operational Classification Unit Community Detection), which divides large-scale metagenomic sequence data into potential gene modules. We construct an OTU network, and then performs subsequent nonoverlapping community detection task with graph convolutional networks. Experimental scores show that the community detection effect of this method is better than other latest metagenomic algorithms.

Recommended nitrogen rates and the verification of effects based on leaf SPAD readings of rice.

Modern rice production systems need a reliable, easy-to-use, efficient, and environmentally-friendly method to determine plant nitrogen (N) status , predict grain yield, and optimize N management. We conducted field experiments to determine the influence of different N rates on Soil Plant Analysis Development (SPAD) readings of rice leaves. We also performed field validations to evaluate the grain yield and N use efficiency under recommended N rates. Our results showed that leaf SPAD readings increased as N rates increased. We applied the recommended N based on the relationships between the N rates and leaf SPAD readings at the tillering and booting stages. The recommended N decreased N rates and improved N use efficiency without sacrificing grain yield. When compared to farmer practices (FP), the recommended N rates of optimization (OPT) decreased by 5.8% and 10.0%, respectively. In comparison with FP, the N agronomic efficiency of OPT increased by 5.8 and 10.0% while the partial factor productivity of N increased by 6.0 and 14.2%, respectively. The SPAD meter may be a reliable tool to analyze the N in rice, estimate real-time N fertilization, and improve N use efficiency.

Preparation and characterization of cornstalk microspheric hydrochar and adsorption mechanism of mesotrione.

In this study, cornstalk was pyrolysed to obtain hydrochar (HC), which was used to remove mesotrione from aqueous solutions. HC characterization and batch experiments were conducted to investigate mesotrione adsorption and the underlying mechanism. The characterization revealed microspheres on the HC surface. FT-IR spectra showed that the HC contained a large number of -OH groups, C=C bonds of aromatic rings, C-H groups in aromatic rings and phenolic C-O bonds. The adsorption results showed that the mesotrione adsorption ability gradually increased as the HC preparation temperature increased. The quasi-second-order kinetic equation (R2 ≥ 0.9860, p < 0.05) agreed well with the mesotrione adsorption process. The maximum monolayer adsorption capacity, which was obtained at pH 7 and 45°C with HC prepared at 240°C, was 3181.7 mg kg-1 with the Langmuir isotherm model (R2 ≥ 0.9491, p < 0.05). Van der Waals and dipole forces and hydrogen bonds were inferred as the main adsorption mechanisms. HC has potential as an effective and energy-saving adsorbent for mesotrione to reduce environmental pollution.

PmDNE: Prediction of miRNA-Disease Association Based on Network Embedding and Network Similarity Analysis.

Successful prediction of miRNA-disease association is nontrivial for the diagnosis and prognosis of genetic diseases. There are many methods to predict miRNA and disease, but biological data are numerous and complex, and they often exist in the form of network. How to accurately use the features of miRNA and disease-related biological networks to predict unknown association has always been a challenge. Here, we propose PmDNE, a method based on network embedding and network similarity analysis, to predict the miRNA-disease association. In PmDNE, the structure of network bipartite graph is improved, and a random walk generator is designed. For embedded vectors, 128 dimensions are used, and the accuracy of prediction is significantly improved. Compared with other network embedding methods, PmDNE is comparable and competitive with the state of art methods. Our method can solve the problem of feature extraction, reduce the dimension of features, and improve the efficiency of miRNA-disease association prediction. This method can also be extended to other area for biomedical network prediction.

The adsorption and mechanism of the nitrification inhibitor nitrapyrin in different types of soils.

The nitrapyrin was easily adsorbed by soil, but most current studies have focused on comparing the effects of nitrapyrin application at different soil organic matter levels and in different soil types. The adsorption kinetics and isotherm adsorption of the nitrification inhibitor nitrapyrin in black soil, chernozem and planosol were studied in this paper. The adsorption kinetics were fitted by quasi-second-order kinetic equation (R2 ≥ 0.8907, p < 0.05) with a lower acting energy of adsorption (Ea < 8.0 kJ mol-1). The isotherm was fitted by the Langmuir equation (R2 ≥ 0.9400*, p < 0.05). The adsorption mechanism was determined to involve a spontaneous endothermic reaction accompanied mainly by physical adsorption to the surface that belonged to the 'L' isotherm curve (n > 1). Temperature promoted the adsorption of nitrapyrin in these three soils, and the maximum adsorption occurring at different temperatures following the order of black soil > planosol > chernozem. The adsorption capacity and rate decreased with decreasing soil organic matter. For the black soil, the nitrapyrin EC adsorption rate was more than seven times higher than that of nitrapyrin CS. The result would determine the dose of nitrapyrin required for availability in different types of soils and to provide a theoretical basis for elucidating the adsorption of nitrapyrin in the soil environment.

Adsorption of nitrification inhibitor nitrapyrin by humic acid and fulvic acid in black soil: characteristics and mechanism.

The compound nitrapyrin is easily adsorbed by soil organic matter in high-organic matter soils, and this results in its effectiveness reducing significantly. In this study, the adsorption characteristics and mechanisms of nitrapyrin as an adsorptive on humic acid (HA) and fulvic acid (FA) as adsorbents were investigated. The results showed that the kinetics of adsorption of nitrapyrin on both HA and FA followed pseudo-second-order kinetic models (R 2 ≥ 0.925, P < 0.05) and the adsorption process included an initial fast-adsorption stage and a slow-adsorption stage thereafter. The adsorption efficiencies of nitrapyrin on HA + FA were higher than that on HA or FA alone, and that of HA was higher than that of FA. The adsorption isotherms of nitrapyrin on HA and FA could be optimally fitted with the Langmuir equation (R 2 ≥ 0.982, P < 0.05). The maximum adsorption capacities of nitrapyrin on HA, FA and HA + FA were 4896.49, 3173.70 and 4925.56 mg kg-1, respectively. Synergistic adsorption of nitrapyrin in co-existing systems of HA and FA was also observed. The adsorption mechanism of nitrapyrin on both HA and FA involved hydrogen bonding and hydrophobic interaction. Therefore, HA and FA in the soil environment can adsorb a large amount of nitrapyrin and reduce its effectiveness, and they have a positive synergistic effect.

Genetic diversities and phylogenetic analyses of three Chinese main ethnic groups in southwest China: A Y-Chromosomal STR study.

Short tandem repeats (STRs) located on the Y chromosome with the properties of male-specific inheritance and haploidy are widely used in forensics to analyze paternal genealogies and match male trace donors to evidence. Besides, Y-chromosomal haplotypes play an important role in providing breathtaking insights into population genetic history. However, the genetic diversity and forensic characteristics of Y-STRs in Guizhou main ethnic groups (Hans, Miaos and Bouyeis) remain uncharacterized. Here, we obtained Y-chromosomal 23-marker haplotypes in three Guizhou populations and submitted the first batch of Y-STR haplotype data to the YHRD. The HD in the aforementioned three populations are 0.99990, 0.99983, and 0.99979, respectively, and DC values are 0.9902, 0.9908, and 0.97959, respectively. Subsequently, genetic differentiation between our newly studied populations and reference groups along ethnic/administrative divisions, as well as national/continental boundaries were investigated via AMOVA, MDS, and phylogenetic relationship reconstruction. Significant genetic differentiations from our subjects and other groups are identified in ethnically, linguistically and geographically diverse populations, including most prominently Tibetans and Uyghurs among 30 mainland Chinese populations, Taiwanese groups and others among 58 Asian populations, as well as African groups and others among 89 worldwide populations. Qiannan Bouyei has a close genetic relationship with Guangxi Zhuang, and Zunyi Han and Qiandongnan Miao have close genetic affinity with Hunan Han and Guizhou Shui, respectively. Collectively, this new-generation Y-STR amplification system can be used as a supplementary tool in forensic identification and male parentage testing and even pedigree search.

[Prosapogenin A inhibits cell growth of MCF7 via downregulating STAT3 and glycometabolism-related gene].

This study is to investigate the inhibitory effect and mechanism of prosapogenin A (PSA) on MCF7. MTT assay was performed to determine the inhibitory effect of PSA on MCF7 cells. PI/Hoechst 33342 double staining was used to detect cell apoptosis. RT-PCR was used to test the mRNA levels of STAT3, GLUT1, HK and PFKL. Western blotting was performed to determine the expression of STAT3 and pSTAT3 protein in MCF7 cells. The results showed that PSA could dose-dependently inhibit cell growth of MCF7 followed by IC50 of 9.65 micrmol x L(-1) and promote cell apoptosis of MCF7. Reduced mRNA levels of STAT3, HK and PFKL were observed in MCF7 cells treated with 5 micromol x L(-1) of PSA. PSA also decreased the level of pSTAT3 protein. STAT3 siRNA caused decrease of mRNA of GLUT1, HK and PFKL which indicated STAT3 could regulate the expressions of GLUT1, HK and PFKL. The results suggested that PSA could inhibit cell growth and promote cell apoptosis of MCF7 via inhibition of STAT3 and glycometabolism-related gene.

Prosapogenin A induces apoptosis in human cancer cells in vitro via inhibition of the STAT3 signaling pathway and glycolysis.

Signal transducer and activator of transcription 3 (STAT3) is considered to be an oncogene. Blocking STAT3 signaling may induce growth arrest and apoptosis in different types of tumors. Cancer cells utilize the glycolytic pathway to maintain cell growth even when adequate oxygen is present. Glycolysis inhibition is a potential therapeutic modality. In the present study, the effects of Prosapogenin A (PSA) from the traditional Chinese medicine, Veratrum, on apoptosis, the STAT3 signaling pathway and glycometabolism in cancer cells were investigated. The results indicated that PSA induced growth inhibition and apoptosis in HeLa, HepG2 and MCF-7 cells. PSA inhibited the STAT3 signaling pathway and modulated the expression of glycometabolism-related genes. The results indicate that the inhibition of the STAT3 signaling and glycometabolism pathways contributes to the PSA-mediated apoptosis of HeLa, HepG2 and MCF-7 cells.

[Small interfering RNA targeting STAT3 enhances antitumor activity of doxorubicin].

This study is to investigate the effect of small interfering RNA targeting STAT3 (STAT3-siRNA) enhancing antitumor activity of doxorubicin. RT-PCR and Western blotting were used to test the expression of STAT3 mRNA and protein in the HepG2, HeLa and K562/DOX cells and the effect of STAT3-siRNA on the expression of STAT3 mRNA and protein. MTT and Trypan blue assay were performed to determine the inhibitory effect of STAT3-siRNA on HepG2, HeLa and K562/DOX cells and the effect of STAT3-siRNA enhancing antitumor activity of doxorubicin. The effects of STAT3-siRNA on intracellular accumulation of doxorubicin and cell apoptosis were performed by Arrary Scan V(TI)HCS600 High-Contents. The results showed that STAT3 gene, STAT3 and pSTAT3 protein were highly expressed in HepG2, HeLa and K562/DOX cells and STAT3-siRNA decreased the expression of STAT3 mRNA and protein. STAT3-siRNA inhibited the growth of HepG2, HeLa and K562/DOX cells. STAT3-siRNA in combination with doxorubicin decreased by 3.13, 5.22 and 1.74 fold of IC50 of HepG2, HeLa and K562/DOX cells compared with doxorubicin only. Intracellular accumulation of doxorubicin increased by 16.8%, 12.87% and 25.67% respectively in HepG2, HeLa and K562/DOX cells in the presence of STAT3-siRNA. An enhancement of doxorubicin-induced cell apoptosis was observed in HepG2, HeLa and K562/DOX cells treated with STAT3-siRNA. The results suggested that STAT3-siRNA could enhance the antitumor activity of doxorubicin on HepG2, HeLa and K562/DOX cells.

[Crop-soil nitrogen cycling and soil organic carbon balance in black soil zone of Jilin Province based on DSSAT model].

By using the CERES-Maize crop model and Century soil model in Decision Support System of Agrotechnology Transfer (DSSAT) model, this paper studied the effects of crop management parameters, fertilizer N application rate, soil initial N supply, and crop residue application on the maize growth, crop-soil N cycling, and soil organic C and N ecological balance in black soil (Mollisol) zone of Jilin Province, Northeast China. Taking 12,000-15,000 kg x hm(-2) as the target yield of maize, the optimum N application rate was 200-240 kg N x hm(-2). Under this fertilization, the aboveground part N uptake was 250-290 kg N x hm(-2), among which, 120-140 kg N x hm(-2) came from soil, and 130-150 kg N x hm(-2) came from fertilizer. Increasing the N application rate (250-420 kg N x hm(-2)) induced an obvious increase of soil residual N (63-183 kg x hm(-2)); delaying the N topdressing date also induced the increase of the residual N. When the crop residue application exceeded 6000 kg x hm(-2), the soil active organic C and N could maintain the supply/demand balance during maize growth season. To achieve the target maize yield and maintain the ecological balance of soil organic C and N in black soil zone of Jilin Province, the chemical N application rate would be controlled in the range of 200-240 kg N x hm(-2), topdressing N should be at proper date, and the application amount of crop residue would be up to 6000 kg x hm(-2).