[Effect of TLK2 Expression Regulated by MiR-21 on Proliferation and Apoptosis of Acute Myeloid Leukemia Cells].

OBJECTIVE: To investigate the effect of TLK2 expression regulated by miR-21 on proliferation and apoptosis of acute myeloid leukemia cells. METHODS: Seventy patients with AML admitted to our hospital from January 2019 to July 2022 were selected, while 30 patients with iron deficiency anemia were selected as the control group. Bone marrow mononuclear cells (BMMNCs) of the patients were obtained using Ficoll density gradient centrifugation. RT-qPCR was used to determine the expression levels of miR-21 and TLK2 mRNA in BMMNCs. Mimics-miR-21, mimics-NC, inhibitor-miR-21, inhibitor-NC and NC were transfected into HL-60 cells using liposome-mediated transfection technology. CCK-8 method was used to determine the activity of transfected HL-60 cells after treatment with cytarabine. The apoptosis rate of HL-60 transfected cells was determined by TUNEL method. The expression of TLK2 mRNA in HL-60 cells transfected with inhibitor-miR-21 was determined by RT-qPCR. RESULTS: The relative expression levels of miR-21 and TLK2 mRNA in BMMNCs of AML patients were significantly higher than those of controls (both P < 0.05). After HL-60 cells were treated with cytarabine, both the cell activity of inhibitor-miR-21 group and mimics-miR-21 group decreased significantly with the increase of cytarabine concentration (both P < 0.05). However, at each concentration point of cytarabine, the cell activity of inhibitor-miR-21 group was lower than that of control group (P < 0.05), while mimics-miR-21 group was higher than control group (P < 0.05). After HL-60 cells were treated with cytarabine, the apoptosis rate of inhibitor-miR-21 group was significantly increased (P < 0.05), while that of mimics-miR-21 group was significantly decreased (P < 0.05). After HL-60 cells were treated with inhibitor-miR-21, the relative expression of TLK2 mRNA decreased significantly (P < 0.05). CONCLUSION: miR-21 is highly expressed in AML patients, which may promote the apoptosis of AML cells by inhibiting the expression of TLK2.

[Effects of Different Water Stratification on the Vertical Distribution of Nitrogen in Sediment Interstitial Waters: A Case Study of the Three Gorges Reservoir and Xiaowan Reservoir].

To determine the reasons for the variation in the vertical distribution of nitrogen in sediment interstitial waters between different stratified reservoirs, the characteristics of overlying water-interstitial water in Xiangxi Bay, Yangtze River mainstream, and Xiaowan Reservoir were monitored. The vertical distribution of nitrogen in sediment interstitial waters in these different stratified waters were then analyzed, and the reasons for the variation in this distribution were assessed. The results showed:① the ρ(TN) in the sediment interstitial waters of the Yangtze River mainstream and Xiangxi Bay gradually increased with depth, while that of Xiaowan Reservoir reached its maximum at 12 cm and the bottom layer presented a "C" distribution. The ρ(NH4+) in the sediment interstitial waters of the Yangtze River mainstream and Xiangxi Bay exhibited an increasing trend with depth, while that of Xiaowan Reservoir was slightly higher in the bottom layer than in the surface layer, although the change with depth was not significant. Overall, the ρ(NH4+) in the sediment interstitial waters of the Yangtze River mainstream and Xiangxi Bay was higher than that of Xiaowan Reservoir, and the concentration ranges were as follows:0.512-8.289 mg·L-1, 0.968-9.307 mg·L-1, and 0.950-1.450 mg·L-1. The vertical distribution of the ρ(NO3-) in the sediment interstitial waters of all three waterbodies were opposite to that of ρ(NH4+). Moreover, the ρ(NO3-) in the sediment interstitial waters of Xiangxi Bay and the Yangtze River mainstream was higher than that of Xiaowan Reservoir. The concentration ranges were as follows:0.143-0.674 mg·L-1, 0.107-0.647 mg·L-1, and 0.050-0.051 mg·L-1. ② There were also significant differences in the vertical distribution of physical and chemical indices in the three water bodies. There was no significant change in the vertical distribution of the water temperature in the Yangtze River mainstream and the N2 value was <5×10-5 s-2; hence, the water was well mixed, and the vertical range of the dissolved oxygen content was 6.180-6.318 mg·L-1. The water temperature in the upper and middle reaches of Xiangxi Bay decreased vertically, while the water temperature in the lower reach presented a ladder-like distribution and the N2 values were all>5×10-5 s-2; thus, the water was in a stable stratified state and the dissolved oxygen content presented a "C" distribution. There was obvious stratification at the depths of 5-15 m and 54-70 m in Xiaowan Reservoir. The dissolved oxygen content decreased significantly at higher water temperature gradients, and there was no significant change along the water depth below 80 m. ③ The main reasons for the variation in the vertical distribution of nitrogen in the sediment interstitial waters of the three waterbodies were the differences in the overlying water hydrodynamics, dissolved oxygen distribution, and sediment environment. The ρ(NH4+) and ρ(NO3-) were higher in Xiangxi Bay, which may have increased the denitrification rate and subsequently have helped to remove nitrogen and reduce the nitrogen load in these waters.

[Analysis of Total Organic Carbon Source Differences Between New and Old Cascade Reservoirs using Carbon and Nitrogen Isotopes].

The ecological problems due to reservoir construction are causing unprecedented concern. To reveal the differences in organic carbon distribution characteristics and sediment sources of total organic carbon (TOC) between the old and new reservoirs, water samples, and sediment samples from reservoirs constructed in the three different periods of Miaowei, Gongguoqiao, and Dachaoshan were collected in November 2017. The temperature (T), dissolved oxygen (DO), TOC, redox potential (ORP), total nitrogen (TN), and total phosphorus (TP) of the water samples were measured. The isotopes 15N and 13C were used as indicators with IsoSource software to analyze the contributions of TOC sources and their source materials to the corresponding reservoir sediments, in order to explore the carbon cycle mechanism and evolution mode of reservoir. The results showed that the average concentrations of organic carbon in the waters of the Miaowei, Gongguoqiao, and Dachaoshan Reservoirs were 0.95 mg·L-1, 1.97 mg·L-1, and 4.64 mg·L-1, respectively. The range of organic carbon content in the corresponding sediments was 4.41-81.63 g·kg-1, 18.30-28.42 g·kg-1, and 9.16-14.46 g·kg-1, respectively. The cascade construction of the reservoirs resulted in a difference between the sediment sources of the new and old reservoirs and the surrounding recharge area, meaning that the TOC of the new and old reservoirs were significantly different. For the TOC of waterbodies, the difference between the thermodynamic state of water and dissolved oxygen indirectly affects the distribution trend of TOC. The sediments mainly reflect the influence of source elements, that is, the ability of the sedimentary environment to preserve organic matter was the main cause of the vertical distribution of DCS, MV, and GGQ sediments. In the evolution mode of cascade reservoir, the research shows that it can be preliminarily set as three stages. Firstly, due to the short age of MV, it is in the first stage and mainly accumulates the TOC from the upstream. GGQ is longer than the age of MV, and it is mainly used to decompose the upstream TOC, so it is defined as in the second stage. Finally, as an old reservoir, DCS mainly accumulates TOC sources around the reservoir, which can be regarded as the third stage.

Glutamate dehydrogenase inhibits tumor growth in gastric cancer through the Notch signaling pathway.

Glutamate dehydrogenase (GDH) is a key enzyme in glutaminolysis and can regulate allosteric functions. Immunohistochemical study found that GDH expressed in gastric cancer cell cytoplasm and membrane, and a few located in the nucleus, ranging from light yellow to tan to sepia. According to the analysis by Kaplan Meier survival curve and the Log-Rank test, the median survival of GDH high expression in patients was 51.7 months with 95% confidence intervals (CI) was 41.138-55.262. The expression level of GDH was significantly reduced after silencing GDH gene in gastric cancer cells and tissues. Further, after silencing GDH gene, gastric cancer cell migration and invasion ability were decreased significantly. Protein expression of. In addition, tumor growth was significantly reduced after silencing GDH gene. In vivo and in vitro experiments suggest that GDH can decrease gastric cancer cell migration and invasion, thus inhibiting tumor growth.

[Speciation and Transformation of Phosphorus in Sediments During the Redox Cycle].

To study the mechanism of phosphorus cycling in sediment during the redox cycle, changes in physicochemical properties of overlying water and various forms of phosphorus in sediments were investigated as a way to quantify the redistribution of phosphorus. Additionally, the effect of the release flux of phosphate from sediments under controlled redox conditions was analyzed. The results showed that the redox potential Eh and the pH system, sulfur system, carbon system, and iron-related changes exhibited periodicity and played an important role in explaining the migration and transformation mechanism in the interface phosphorus of the sediment-water phase. During the redox cycle, the phosphorus content of each species varied with the redox conditions and time. Because of this, quantitative analysis based on changes in water-sediment phosphorus could be obtained. Reducible phosphorus (BD-P) and iron-aluminum-bound phosphorus (NaOH-rP) were reversibly redistributed into weakly adsorbed phosphorus (NH4Cl-P), polyphosphorus/organophosphorous (NaOH-nrP), residual phosphorus (Rest-P), and interstitial water-soluble active phosphorus (SRP). Additionally, 93.7% of phosphorus in the sediment was not released into the water phase during the reduction reaction. The 92% of change in the overlying water total phosphorus (TP) was the SRP of overlying water, which showed that the exchange of the sediment-water phase were mainly soluble active phosphorus in this cycle. According to Fick's First Law, the maximum phosphorus flux was 0.58 mg·(m2·d)-1 during reduction and 0.16-0.22 mg·(m2·d)-1 on day seven of the oxidation phase. In the oxidation stage, the diffusion flux decreased with time, while the opposite trend occurred in the reduction reaction. This indicated that the anaerobic state accelerated the diffusion of phosphorus in sediments, and that oxygen exposure decreased the phosphorus flux in sediments.

Ultrasound-guided percutaneous microwave ablation vs. surgical resection for thoracoabdominal wall implants from hepatocellular carcinoma: intermediate-term results.

OBJECTIVE: The aims of this study were to compare the clinical outcomes between ultrasound (US)-guided percutaneous microwave ablation (MWA) and surgical resection (SR) in patients with thoracoabdominal wall implants from hepatocellular carcinom (HCC) and to identify the prognostic factors associated with the two treatment methods. MATERIALS AND METHODS: A total of 47 patients (mean age, 56.7 ± 15.9 years, range, 18-78 years; 34 men and 13 women) with 61 thoracoabdominal wall HCC seeding were included from April 2007 to May 2017. Twenty-five patients underwent US-guided MWA and 22 patients underwent SR. Survival, recurrence and liver function were compared between the two groups. Effect of changes in key parameters (i.e. overall survival (OS), disease-free survival (DFS) and local tumour reoccurrence-free (LTRF)) was statistically analysed with the log-rank test. Univariate and multivariate analyses were performed on several clinicopathological variables to identify factors affecting long-term outcome and recurrence. RESULTS: The OS, DFS and LTRF after MWA were comparable to those of SR (p =0.493, p = 0.578 and p =0.270, respectively). Estimated 5-year overall survival rates were 63% after MWA and 48.1% after SR; for disease-free survival, estimated 5-year rates were 67.5% after MWA and 48.8% after SR; estimated 24-month LTRF rates were 71.3% after MWA and 87.8% after SR. The MWA group had less surgical time (p = <0.001), estimated blood loss (p = <0.001) and post-operative hospitalisation (p = 0.032) and cost (p = 0.015). Multivariate analysis showed remnant intrahepatic tumour (p =0.007), Child Pugh grade (p = 0.009) and metastasis (p= <0.001), were predictors for survival rate. CONCLUSIONS: Ultrasound-guided percutaneous MWA is a safe and effective treatment method for metastatic HCC on the thoracoabdominal wall with similar outcomes to SR. Residual intrahepatic HCC, Child Pugh grade and distant metastasis are predictors for survival.

Metabolic alterations in the rat cerebellum following acute middle cerebral artery occlusion, as determined by 1H NMR spectroscopy.

Supratentorial focal ischemia may reduce cerebral blood volume and cerebellar glucose metabolic rate contralateral to the region of ischemia. The present study investigated the effects of middle cerebral artery occlusion (MCAO) on cerebral metabolism in the ischemic cerebral hemisphere and the non­ischemic cerebellum in rats 1, 3, 9 and 24 h following ischemia using ex vivo proton nuclear magnetic resonance (1H NMR) spectroscopy. The results demonstrated that focal ischemia induced increases in the levels of lactate and alanine, and a decrease in succinate, as early as 1 h following ischemia in the left cerebral hemisphere and the right cerebellum. A continuous increase in lactate levels and decrease in creatine levels were detected in both cerebral areas 3 and 24 h post­MCAO. The most obvious difference between the two cerebral areas was that there was no statistically significant difference in N­acetyl aspartate (NAA) levels in the right cerebellum at all time points; however, the amino acid levels of NAA in the left cerebral hemisphere were markedly decreased 3, 9 and 24 h post­MCAO. In addition, an obvious increase in glutamine was observed in the right and left cerebellum at 3, 9 and 24 h post­MCAO. Furthermore, the present study demonstrated that γ­aminobutyric acid levels were decreased at 1 h in the left and right cerebellum and were evidently increased at 24 h in the right cerebellum post­MCAO. In conclusion, supratentorial ischemia has been indicated to affect the activities of the non­ischemic contralateral cerebellum. Therefore, these results suggested that an NMR­based metabonomic approach may be used as a potential means to elucidate cerebral and cerebellar metabolism following MCAO, which may help improve understanding regarding cerebral infarction at a molecular level. Ex vivo 1H NMR analysis may be useful for the assessment of clinical biopsies.

Metabolite changes in the ipsilateral and contralateral cerebral hemispheres in rats with middle cerebral artery occlusion.

Cerebral ischemia not only causes pathological changes in the ischemic areas but also induces a series of secondary changes in more distal brain regions (such as the contralateral cerebral hemisphere). The impact of supratentorial lesions, which are the most common type of lesion, on the contralateral cerebellum has been studied in patients by positron emission tomography, single photon emission computed tomography, magnetic resonance imaging and diffusion tensor imaging. In the present study, we investigated metabolite changes in the contralateral cerebral hemisphere after supratentorial unilateral ischemia using nuclear magnetic resonance spectroscopy-based metabonomics. The permanent middle cerebral artery occlusion model of ischemic stroke was established in rats. Rats were randomly divided into the middle cerebral artery occlusion 1-, 3-, 9- and 24-hour groups and the sham group. 1H nuclear magnetic resonance spectroscopy was used to detect metabolites in the left and right cerebral hemispheres. Compared with the sham group, the concentrations of lactate, alanine, γ-aminobutyric acid, choline and glycine in the ischemic cerebral hemisphere were increased in the acute stage, while the concentrations of N-acetyl aspartate, creatinine, glutamate and aspartate were decreased. This demonstrates that there is an upregulation of anaerobic glycolysis (shown by the increase in lactate), a perturbation of choline metabolism (suggested by the increase in choline), neuronal cell damage (shown by the decrease in N-acetyl aspartate) and neurotransmitter imbalance (evidenced by the increase in γ-aminobutyric acid and glycine and by the decrease in glutamate and aspartate) in the acute stage of cerebral ischemia. In the contralateral hemisphere, the concentrations of lactate, alanine, glycine, choline and aspartate were increased, while the concentrations of γ-aminobutyric acid, glutamate and creatinine were decreased. This suggests that there is a difference in the metabolite changes induced by ischemic injury in the contralateral and ipsilateral cerebral hemispheres. Our findings demonstrate the presence of characteristic changes in metabolites in the contralateral hemisphere and suggest that they are most likely caused by metabolic changes in the ischemic hemisphere.

Alteration of interaction between astrocytes and neurons in different stages of diabetes: a nuclear magnetic resonance study using [1-(13)C]glucose and [2-(13)C]acetate.

Increasing evidence has shown that the brain is a site of diabetic end-organ damage. This study investigates cerebral metabolism and the interactions between astrocytes and neurons at different stages of diabetes to identify the potential pathogenesis of diabetic encephalopathy. [1-(13)C]glucose or [2-(13)C]acetate is infused into 1- and 15-week diabetic rats, the brain extracts of which are analyzed by using (1)H and (13)C magnetic resonance spectroscopy. The (13)C-labeling pattern and enrichment of cerebral metabolites are also investigated. The increased (13)C incorporation in the glutamine, glutamate, and γ-aminobutyric acid carbons from [2-(13)C]acetate suggests that the astrocytic mitochondrial metabolism is enhanced in 1-week diabetic rats. By contrast, the decreased labeling from [1-(13)C]glucose reflected that the neuronal mitochondrial metabolism is impaired. As diabetes developed to 15 weeks, glutamine and glutamate concentrations significantly decreased. The increased labeling of glutamine C4 but unchanged labeling of glutamate C4 from [2-(13)C]acetate suggests decreased astrocyte supply to the neurons. In addition, the enhanced pyruvate recycling pathway manifested by the increased lactate C2 enrichment in 1-week diabetic rats is weakened in 15-week diabetic rats. Our study demonstrates the overall metabolism disturbances, changes in specific metabolic pathways, and interaction between astrocytes and neurons during the onset and development of diabetes. These results contribute to the mechanistic understanding of diabetes pathogenesis and evolution.