CARMIL1 regulates liver cancer cell proliferation by activating the ERK/mTOR pathway through the TRIM27/p53 axis.

Capping protein regulatory factor and myosin 1 linker 1 is termed CARMIL1. CARMIL1 is involved in several physiological processes; it forms an actin filament network and plasma membrane-bound cellular projection tissues and positively regulates the cellular components and tissues. CARMIL1 exhibits important biological functions in cancer; nonetheless, these functions have not been completely explored. We aimed to investigate the novel functions of CARMIL1 in liver cancer, particularly in cell proliferation. The cell counting kit-8, 5-ethynyl-2'-deoxyuridine, Component A experiments, and subcutaneous tumor formation model suggest that CARMIL1 is central to the proliferation of liver cancer cells both in vivo and in vitro. We extracted CARMIL1 samples from The Cancer Genome Atlas Program and analyzed its enrichment. CARMIL1 regulated the pathway activity by affecting the expression of star molecular proteins of the extracellular signal-regulated kinase (ERK) and mammalian target of rapamycin (mTOR). Moreover, it influenced the proliferation ability of liver cancer cells. Western blotting suggested that CARMIL1 downregulation could affect ERK and mTOR phosphorylation. Results of the co-immunoprecipitation demonstrated that CARMIL1 binds to tripartite motif (TRIM)27, which in turn binds to p53. Subsequently, CARMIL1 can regulate p53 stability and promote its degradation through TRIM27. Additionally, CARMIL1 inhibition enhanced the sensitivity of liver cancer cells to sorafenib. Tumor growth was significantly inhibited in the group treated with sorafenib and CARMIL1, compared with the group treated with CARMIL1 alone. Sorafenib is a first-line targeted chemotherapeutic drug for hepatocellular carcinoma treatment. It increases the long-term survival of hepatocellular carcinoma by 44%. In this study, downregulated CARMIL1 combined with sorafenib significantly reduced the tumor volume and weight of the mouse subcutaneous tumor model, indicating the potential possibility of combining CARMIL1 with sorafenib in hepatocellular carcinoma treatment. In summary, CARMIL1 promotes liver cancer cell proliferation by regulating the TRIM27/p53 axis and activating the ERK/mTOR pathway.

Effects of 2-dodecyl-6-methoxycyclohexa-2,5-diene-l ,4-dione on resisting hepatic fibrosis induced by CC14 in rats and its mechanisms via TGF-pi/Smads signaling pathway / 中国药理学通报

Aim To investigate the effects of 2-dode-cyl-6-methoxycyclohexa-2 , 5-diene-l, 4-dione ( DM-DD) on resisting hepatic fibrosis induced by carbon tetrachloride ( CC14 ) in rats and the underlying mechanisms , with a specific focus on the TGF-pi/Smads signaling pathway. Methods The hepatic fibrosis model was replicated using 50% CC14. Various parameters, including levels of aspartate transferase ( AST) , ala-nine transferase ( ALT ) , albumin/globulin ( A/G ) , total protein (TP) , total bilirubin (T-BIL) , hyaluron-ic acid ( HA ) , laminin ( LN ) , collagen type Ж ( Col Ж) , and collagen type IV(ColIV) in the blood, were measured. Liver tissue lesions and fiber formation were observed using HE and Masson staining. The expression levels of a smooth muscle actin (a-SMA) , collagen type I ( Col I ) , transformed growth factor (TGF-pi), Smad2, and Smad7 proteins were assessed using immunohistochemistry. a-SMA, Coll, TGF-pi, and Smad7 mRNA levels in liver tissue were measured by RT-PCR. Additionally, the expression levels of TGF-pi, Smad4, and Smad7 proteins in liver tissue were determined by Western blot. Results In comparison to the normal control group, the model group exhibited significantly elevated levels of AST, ALT, TP, T-BIL, HA, LN, Col Ш and Col IV in serum. But A/G level notably decreased. Successful modeling was confirmed by the presence of extensive fiber formations observed through HE and Massonstaining in liver tissue. The DMDD administration group demonstrated a notable decrease levels of AST, ALT, TP, T-BIL, HA, LN, Col III, and CollV, but A/G was significantly elevated when compared to the model group. Furthermore, a-SMA, Coll, TGF-f31, Smad2 and Smad4 mRNA and protein levels in the DMDD administration group were significantly reduced, while Smad7 significantly declined. HE and Masson staining results reflected a marked reduction in fibrous hyper-plasia. Conclusion DMDD exhibits a protective effect against CCl4-induced hepatic fibrosis, and its mechanism appears to be associated with the TGF-fJl/ Smads signaling pathway.

CDN1163, a SERCA activator, causes intracellular Ca2+ leak, mitochondrial hyperpolarization and cell cycle arrest in mouse neuronal N2A cells.

OBJECTIVE: Activity or expression of sarcoplasmic/endoplasmic reticulum Ca2+ ATPase (SERCA) is diminished in some disease states such as cardiac failure and diabetes mellitus. A newly developed activator of SERCA, CDN1163, reportedly rescued or alleviated pathological conditions attributed to dysfunctional SERCA. We examined whether CDN1163 could relieve mouse neuronal N2A cell growth inhibition caused by cyclopiazonic acid (CPA, SERCA inhibitor). We also examined how CDN1163 affected cytosolic Ca2+, mitochondrial Ca2+ and mitochondrial membrane potential. METHODS: Cell viability was measured by MTT assay and trypan blue exclusion test. Cytosolic Ca2+, mitochondrial Ca2+ and mitochondrial membrane potential were measured using fura 2, Rhod-2 and JC-1, respectively, as fluorescent probes. RESULTS: CDN1163 (10 µM) itself suppressed cell proliferation, and did not alleviate CPA's inhibitory effect (and vice versa). Cell cycle was arrested at the G1 phase after CDN1163 treatment. CDN1163 treatment caused a slow yet persistent cytosolic [Ca2+] elevation partly due to Ca2+ release from an internal store other than the CPA-sensitive endoplasmic reticulum (ER). Treatment with CDN1163 for 3 h raised mitochondrial Ca2+ level and such increase was suppressed by MCU-i4 (an inhibitor of mitochondria Ca2+ uniporter, MCU), suggesting Ca2+ entered the mitochondrial matrix through MCU. Treatment of cells with CDN1163 up to 2 days resulted in mitochondrial hyperpolarization. CONCLUSION: CDN1163 caused internal Ca2+ leak, cytosolic Ca2+ overload, mitochondrial Ca2+ elevation and hyperpolarization, cell cycle arrest and cell growth inhibition.

A dramatic decline in fruit citrate induced by mutagenesis of a NAC transcription factor, AcNAC1.

Citrate is a common primary metabolite which often characterizes fruit flavour. The key regulators of citrate accumulation in fruit and vegetables are poorly understood. We systematically analysed the dynamic profiles of organic acid components during the development of kiwifruit (Actinidia spp.). Citrate continuously accumulated so that it became the predominate contributor to total acidity at harvest. Based on a co-expression network analysis using different kiwifruit cultivars, an Al-ACTIVATED MALATE TRANSPORTER gene (AcALMT1) was identified as a candidate responsible for citrate accumulation. Electrophysiological assays using expression of this gene in Xenopus oocytes revealed that AcALMT1 functions as a citrate transporter. Additionally, transient overexpression of AcALMT1 in kiwifruit significantly increased citrate content, while tissues showing higher AcALMT1 expression accumulated more citrate. The expression of AcALMT1 was highly correlated with 17 transcription factor candidates. However, dual-luciferase and EMSA assays indicated that only the NAC transcription factor, AcNAC1, activated AcALMT1 expression via direct binding to its promoter. Targeted CRISPR-Cas9-induced mutagenesis of AcNAC1 in kiwifruit resulted in dramatic declines in citrate levels while malate and quinate levels were not substantially affected. Our findings show that transcriptional regulation of a major citrate transporter, by a NAC transcription factor, is responsible for citrate accumulation in kiwifruit, which has broad implications for other fruits and vegetables.

Effects of ant nesting on seasonal dynamics of soil N2O emission in a secondary tropical forest.

We assessed the seasonal dynamics of N2O emission in ant nests soils in secondary tropical Millettia leptobotrya forest of Xishuangbanna by using the static chamber-gas chromatography method, and determined the lin-kages between ant-mediated changes in soil properties (e.g., carbon pool, nitrogen pool, and temperature and humidity) and N2O emission. The results showed that ant nesting significantly affected soil N2O emission. The ave-rage soil N2O emission (0.67 mg·m-2·h-1) in ant nests was 40.2% higher than that in the control (0.48 mg·m-2·h-1). N2O emission in ant nests and the control showed substantial seasonal variation, with higher rate in June (0.90 and 0.83 mg·m-2·h-1, respectively) than that in March (0.38 and 0.19 mg·m-2·h-1, respectively). Ant nesting significantly increased the values (7.1%-74.1%) of moisture, temperature, organic carbon, total nitrogen, hydrolytic nitrogen, ammonium nitrogen, nitrate nitrogen, and microbial biomass carbon, but decreased pH (9.9%) compared with the control. Results of structural equation model showed that soil N2O emission was promoted by soil C and N pool, temperature, and humidity, but was inhibited by soil pH. The explained extents of soil nitrogen pool, carbon pool, temperature and humidity, and pH for N2O emission changes were 37.2%, 27.7%, 22.9% and 9.4%, respectively. Therefore, ant nesting regulated N2O emission dynamics by changing nitrification and denitrification substrates (e.g., nitrate and ammoniacal nitrogen), carbon pool, and micro-habitat (temperature and moisture) of soil in the secondary tropical forest.

[Spatiotemporal Distribution and Pollution Risk Assessment of Heavy Metals in Sediments of Main Water Supply Reservoirs in Central Zhuhai City].

The Modaomen and Hongwan waterways are the main estuaries of the Pearl River system. Affected by tides, the Fenghuangshan Reservoir, Dajingshan Reservoir, and Zhuxiandong Reservoir, which are subject to water transfer along the river line, are important drinking water sources in the central part of Zhuhai City in the Guangdong-Hong Kong-Macao Greater Bay Area. Eight heavy metals in 15 sampling points in the reservoir area were monitored in the two seasons (August and October) of 2019. The environmental quality of the surface sediments of the three water supply reservoirs was surveyed using correlation analysis, geoaccumulation index evaluation, ecological risk assessment, and traceability analysis in the present study. The results showed that the order of the average content of the eight heavy metals in the three reservoirs was as follows: Zhuxiandong Reservoir>Dajingshan Reservoir>Fenghuangshan Reservoir. The contents of Hg, Zn, and Pb in Fenghuangshan Reservoir; Hg, As, Mn, Ni, Cu, Zn, and Pb in Dajingshan Reservoir; and Zhuxiandong Reservoir exceeded the background values of soil heavy metals in Guangdong province by 1.88-2.75, 1.05-2.32, and 1.69-6.45 times. Except for As in Fenghuangshan Reservoir, As and Cu in Dajingshan Reservoir and Cr and Ni in Zhuxiandong Reservoir, the other heavy metals elements showed the content characteristics of dry season>wet season. The evaluation results of the geoaccumulation index method indicated that the pollution degree of the three reservoirs was in the order of: Zhuxiandong Reservoir>Dajingshan Reservoir>Fenghuangshan Reservoir. Among them, the Fenghuangshan Reservoir and the Dajingshan Reservoir were only slightly polluted by Hg, Zn, and Pb elements, and the Zhuxiandong Reservoir as a whole was slightly or moderately polluted. The evaluation results of the potential ecological risk index showed that the overall ecological risk of Fenghuangshan Reservoir and Dajingshan Reservoir was low, and Zhuxiandong Reservoir was at a medium ecological risk level. Hg was the main ecological risk-contributing factor of the three reservoirs, with contribution rates of 78.8%, 64.4%, and 51.0%, respectively.

Endoscopic full-thickness resection with retroperitoneal dissection for duodenal myogenic cyst with adjustable traction from an independently controlled snare.

Video 1Endoscopic full thickness resection with retroperitoneal dissection for duodenal myogenic cyst with adjustable traction from an independently controlled snare.

Detection of early-universe gravitational-wave signatures and fundamental physics.

Detection of a gravitational-wave signal of non-astrophysical origin would be a landmark discovery, potentially providing a significant clue to some of our most basic, big-picture scientific questions about the Universe. In this white paper, we survey the leading early-Universe mechanisms that may produce a detectable signal-including inflation, phase transitions, topological defects, as well as primordial black holes-and highlight the connections to fundamental physics. We review the complementarity with collider searches for new physics, and multimessenger probes of the large-scale structure of the Universe.

Targeting 4-1BB for tumor immunotherapy from bench to bedside.

Immune dysfunction has been proposed as a factor that may contribute to disease progression. Emerging evidence suggests that immunotherapy aims to abolish cancer progression by modulating the balance of the tumor microenvironment. 4-1BB (also known as CD137 and TNFRS9), a member of tumor necrosis factor receptor superfamily, has been validated as an extremely attractive and promising target for immunotherapy due to the upregulated expression in the tumor environment and its involvement in tumor progression. More importantly, 4-1BB-based immunotherapy approaches have manifested powerful antitumor effects in clinical trials targeting 4-1BB alone or in combination with other immune checkpoints. In this review, we will summarize the structure and expression of 4-1BB and its ligand, discuss the role of 4-1BB in the microenvironment and tumor progression, and update the development of drugs targeting 4-1BB. The purpose of the review is to furnish a comprehensive overview of the potential of 4-1BB as an immunotherapeutic target and to discuss recent advances and prospects for 4-1BB in cancer therapy.

[Structural Characteristics of Phytoplankton Communities and Its Relationship with Environmental Factors in a Group of Drinking Water Reservoirs by Water Transmission from Modaomen Waterway in Zhuhai].

Modaomen Waterway is the main outlet of the Pear River system and an important water source for Zhuhai and Macao. The water quality of 13 sampling sites in Modaomen Waterway, phytoplankton, and environmental factors were investigated at 21 sampling sites in 4 drinking water source groups, which transport water from Modaomen Waterway and connect with each other, in August and October of 2021. A total of 73 genera of phytoplankton in eight phyla were identified, with a total of 150 species, most of them belonging to Cyanophyta, Chlorophyta, and Bacillariophyta. The relative abundance of Cyanophyta was higher than 90% in the DJS and YL reservoirs during flood and dry seasons; Cyanophyta, Chlorophyta, and Bacillariophyta were dominant phyla during different seasons; and their distribution was balanced in the ZXD and ZY reservoirs. The Shannon-Wiener index, Pielous uniformity index, and Margalef richness index showed that the ZY reservoir had the most abundant biodiversity, which indicated the best water quality, followed by the ZXD and YL reservoirs. Conversely, the DJS was the least diverse reservoir for phytoplankton. The PCoA analysis indicated significant differences in plankton structures in ZXD and ZY with the other two reservoirs (P<0.05), respectively. Redundancy analysis (RDA) showed that the main environmental factors affecting the distribution of the phytoplankton community were NO3-, TOC, TP, Cl-, and NH4+-N. These results indicated that the phytoplankton community of the four reservoirs were greatly affected by the nutrient salt caused by water transport, which suggested that the water quality of Modaomen Waterway should be improved to increase reservoir water nutrition to ensure the safety of drinking water sources.

Incidence of Hip Fractures among Patients with Chronic Otitis Media: The Real-World Data.

Chronic otitis media (COM) has been considered as a localized disease, and its systemic impact is poorly understood. Whether COM-induced inflammation could be associated with systemic bone loss and hip fracture is unknown at present. Our study tried to determine the risk of hip fracture among COM patients. We selected the comparison individuals without the COM coding and paired the controls with COM patients by gender, age, and comorbidities (including osteoporosis) by about a one-to-two ratio. Our study showed that the incidence of hip fracture was 4.48 and 3.92 per 1000 person-years for comparison and COM cohorts respectively. The cumulative incidence of hip fracture is higher in the COM cohort (p < 0.001). After adjustment for gender, age, and comorbidities, the COM patients had a 1.11-fold (aHR = 1.11; 95% CI = 1.05−1.17) risk of hip fracture than the control subjects. Among COM patients, a history of hearing loss is associated with higher (aHR = 1.21; 95% CI = 1.20−1.42) fracture risk. Our study showed that COM patients, especially those with hearing loss, are susceptible to a higher risk for hip fracture.

Novel Colorectal Endoscopic Submucosal Dissection With Double-Endoscope and Snare-Based Traction.

BACKGROUND: Colorectal endoscopic submucosal dissection is technically demanding, and the traction offered by gravity, cap, or clip-with-line during conventional endoscopic submucosal dissection remains unsatisfactory. Robotic systems are still under development and are expensive. We proposed double-scope endoscopic submucosal dissection with strong and adjustable traction offered by snaring the lesion with additional scope. OBJECTIVE: This study aimed to test the novel double-scope endoscopic submucosal dissection with snare-based traction. DESIGN: This was a retrospective study that reviewed double-scope endoscopic submucosal dissection compared with matched conventional endoscopic submucosal dissection, and size, location, morphology, and pathology between groups were compared. SETTINGS: This study was conducted in a referral endoscopy center in a local hospital. PATIENTS: This study included patients with colorectal lesions receiving double-scope endoscopic submucosal dissection and matched conventional endoscopic submucosal dissection. MAIN OUTCOME MEASURES: The pathological completeness, procedure time, and complications were analyzed. RESULTS: Fifteen double-scope endoscopic submucosal dissection procedures, with 11 lesions located in the proximal colon with a median size of 40 mm, were performed. The median procedure time of double-scope endoscopic submucosal dissection was 32.45 (interquartile range, 16.03-38.20) minutes. The time required for second scope insertion was 2.57 (interquartile range, 0.95-6.75) minutes; for snaring, 3.03 (interquartile range, 2.12-6.62) minutes; and for actual endoscopic submucosal dissection, 28.23 (interquartile range, 7.90-37.00) minutes. All lesions were resected completely. No major complication was encountered. The procedure time was significantly shorter than that of 14 matched conventional endoscopic submucosal dissections (54.61 [interquartile range, 33.11-97.25] min; p = 0.021). LIMITATIONS: This was a single-center, single-operator, retrospective case-controlled study with limited cases. CONCLUSIONS: This study confirmed the feasibility of double-scope endoscopic submucosal dissection with snare-based traction to shorten procedure time and to simplify endoscopic submucosal dissection. Additional trials are required.

[Pollution Characteristics and Risks of Polycyclic Aromatic Hydrocarbons in Underground and Surface Drinking Water Sources in Northeast Inner Mongolia].

Polycyclic aromatic hydrocarbons (PAHs) are hazardous and ubiquitous pollutants in the aquatic environment, and understanding the pollution characteristics and risk levels of PAHs is of great significance to the sustainable development of drinking water sources and drinking water safety. Hence, PAHs residues were measured qualitatively and quantitatively with solid-phase extraction-gas chromatography-mass spectrometry (SPE-GC-MS) in 33 water samples (including 22 groundwater and 11 surface water samples) of the drinking water sources in the Manzhouli and Xinyouqi areas of northeast Inner Mongolia, and assessments of the pollution level of PAHs and the health and ecological risks were carried out. The results showed that PAHs were detected in all 33 sampling points of Manzhouli drinking water sources, except for benzo[k] fluoranthene, benzo[a] pyrene, and dibenzo[a,h] anthracene, with detection rates ranging from 36.36% to 95.45%; the detection rates of the other 13 PAHs monomers were 100%. The detection range of ρ（ΣPAHs） was 42.76-164.50 ng·L-1, and the mean value was 90.82 ng·L-1. The detection ranges of ρ（ΣPAHs） in surface water and groundwater were 66.39-164.50 ng·L-1 and 42.76-147.70 ng·L-1, respectively. The concentration of the detected naphthalene was the highest, with a mean value of 36.91 ng·L-1, and the concentration of anthracene was the lowest, with a mean value of 0.81 ng·L-1; there were no significant differences among the concentrations of all the PAHs monomers of the surface and groundwater (P>0.05). The pollution of PAHs was at a median level in China and abroad, mainly in the middle and low loops (3-4 loops). The analysis of the sources of PAHs in groundwater and surface water in Manzhouli using the ratio feature method and principal component analysis showed that the PAHs in the drinking water source water bodies in the Manzhouli area were mainly affected by the combustion of coal and biomass and oil, and some surface water sources were affected by the oil source. The human health and ecological risk assessment results showed that the water body of drinking water would not cause health risks to the human body, and the ecological risk was at a medium level; however, the high risk of benzo[b] fluoranthene (BbF) monomer production should be continuous cause for concern. From the perspective of the sustainable development of drinking water sources and drinking water safety, the necessary supervision and protection measures should be considered to prevent further pollution. The results of this research provide a scientific basis for the pollution control and prevention and control of PAHs in drinking water sources.

Investigating the effects of travel lane configuration and lane width on traffic safety where powered-two-wheelers (PTWs) share with larger vehicles: A micro perspective.

The design of travel lane configuration and lane width is crucial to traffic safety, especially in an urban mixed traffic environment where Powered-Two-Wheelers (PTWs) are prevalent and share the same roads with larger vehicles such as cars, buses, and trucks. However, there have been limited studies on the effects of the design of travel lane configuration and lane width on safety in such a mixed traffic environment. It's true the above-mentioned research question can be evaluated simply in terms of the number of crashes. However, doing so not only requires a few years of crash and traffic data, but limited insight can be gained in terms of how driver and rider behaviours are affected, and this has implications for further improvement in road safety. This study examines the changes in driving/riding behaviours and surrogate events before and after the adjustments of travel lane configuration and lane width by proposing a micro perspective approach as a complement to conventional studies. A before-and-after site-based investigation was conducted at two study sites which had opposite adjustments for travel lane configuration and lane widths: at one site the number of lanes was reduced, thereby widening the lane width in the outer lane on one road section, and at the second site the number of lanes was increased, thereby narrowing lane width in the outer lane on the other road section. The results showed that an increase in lane width resulted in a considerable increase in the number of speeding events as well as unsafe driving/riding behaviours and surrogate events related to lane splitting, lane sharing, and overtaking.