Objective To observe the effects and safety of adrenocorticotropic hormone (ACTH) combined with Huaiqihuang on frequent relapse nephrotic syndrome (FRNS) in children. Methods Fifty-five child patients with FRNS were divided into control group, which was given glucocorticoid (GC) to maintain the treatment (group A, n=10), Huaiqihuang group (group B, n=17), ACTH group (group C, n=14) and ACTH combined with Huaiqihuang group (combined treatment group, group D, n=14). Continuous treatment was for 12 months. The GC treatment doses, the levels of basal secretion of adrenal cortex and adrenal cortex reserve were recorded at 6-month and 12-month respectively. And the recurrence rate and adverse reactions were observed in four groups. Results After 6-month treatment, the doses of GC were significantly lower in group C and group D than those in group A and group B (P<0.05). The levels of basal secretion of adrenal cortex were increased in turn in group A~D (P<0.05). After 12-month treatment, the doses of GC were significantly decreased in group C and group D than those in group A and group B, while the level of basal secretion of adrenal cortex and adrenal cortex reserve were increased (P<0.05). There were no significant differences in the doses of GC between group C and group D (P>0.05). After treatment for 6 months and 12 months, the recurrence rates of nephrotic syndrome were significantly lower in group C and group D than those of group A and group B (P<0.05). Conclusion The simple application of ACTH and the combination of Huaiqihuang can relieve the inhibition of long-term using GC on hypothalamic pituitary adrenal axis in FRNS patients.

Objective To evaluate effect of dexmedetomidine on mitochondrion-dependent apoptosis during hypoxia-reoxygenation (H/R) injury to hippocampal neurons of rats.Methods The primarily cultured hippocampal neurons of Sprague-Dawley rats were divided into 4 groups (n =40 each) using a random number table method:control group (C group),vehicle group (V group),H/R group and dexmedetomidine group (D group).Hippocampal neurons were subjected to oxygen-glucose deprivation followed by restoration of oxygen supply to establish the model of H/R injury.Dexmedetomidine 1 μmol/L was added at 6 h of reoxygenation in D group.The viability of neurons was measured by methyl thiazolyl tetrazolium assay at 20 h of reoxygenation.The ultrastructure of mitochondria was observed by transmission electron microscopy.The expression of cytochrome c (Cyt c),caspase-3,Fis1 and Drp1 was detected by Western blot.The neuronal apoptosis was detected by flow cytometry,and apoptosis rate was calculated.Results Compared with C group,no significant change was found in the viability of neurons in group V (P＞0.05),and the viability of neurons was significantly decreased,the apoptosis rate was increased,the expression of Cyt c,caspase-3,Fis1 and Drp1 was up-regulated (P＜0.05),and the damage to mitochondrial ultrastructure was accentuated in H/R and D groups.Compared with H/R group,the viability of neurons was significantly increased,the apoptosis rate was decreased,the expression of Cyt c,caspase-3,Fis1 and Drp1 was down-regulated (P＜0.05),and the damage to mitochondrial ultrastructure was significantly attenuated in D group.Conclusion The nechanism by which dexmedetomidine reduces the H/R injury to hippocampal neurons is related to inhibiting mitochondrion-dependent apoptosis in rats.

Objective:To investigate the role of receptor-interacting protein kinse3 (RIPK3)-mediated necroptosis in diabetic mellitus-caused abolition of cardioprotection induced by sevoflurane postconditioning in rats.Methods:Eighty rats with diabetes mellitus, aged 4-5 weeks, weighing 90-100 g, were divided into 4 groups ( n=20 each) using a random number table method: sham operation group (group Sham), myocardial ischemia-reperfusion (I/R) group (group I/R), sevoflurane postconditioning group (group SP) and sevoflurane postconditiong plus RIPK3 inhibitor GSK-872 group (group GSK). Myocardial I/R was induced by 40 min occlusion of the left anterior descending branch of the coronary artery followed by 120 min reperfusion.In group SP, 2.4% sevoflurane was inhaled for 15 min at the beginning of reperfusion.In group GSK, GSK-872 3.3 mg/kg (dissolved in normal saline) was intraperitoneally injected at 24 and 2 h before surgery, and the other treatments were similar to those previously described in group SP.After 120 min of reperfusion, blood samples from the abdominal aorta were collected for determination of concentrations of serum lactate dehydrogenase (LDH) and creatine kinase-MB (CK-MB). Myocardial tissues were taken for determination of percentage of myocardial infarct size (by TTC staining) and expression of RIPK3, phospho-Ca 2+ -calmodulin-dependent protein kinase Ⅱ (p-CaMKⅡ) and phospho-mixed lineage kinase domain-like protein (p-MLKL) (by Western blot), and the ultrastructure of myocardium was observed by transmission electron microscopy. Results:Compared with group Sham, the serum LDH and CK-MB concentrations and percentage of myocardial infarct size were significantly increased, the expression of RIPK3, p-MLKL and p-CaMKⅡ in myocardial tissues was up-regulated ( P<0.05), and the damage to cardiomyocytes was severe in group I/R.Compared with group I/R, no significant change was found in the parameters mentioned above in group SP ( P>0.05). Compared with group SP, the serum LDH and CK-MB concentrations and percentage of myocardial infarct size were significantly decreased, the expression of RIPK3, p-MLKL and p-CaMKⅡ in myocardial tissues was down-regulated ( P<0.05), and the damage to cardiomyocytes was reduced in group GSK. Conclusion:The mechanism of diabetic mellitus-caused abolition of cardioprotection induced by sevoflurane postconditioning is related to excessive activation of RIPK3-mediated necroptosis in rats.

Objective:To study the antitumor activity of zeylenone in colorectal cancer and its related regulatory mechanism.Methods:Human colorectal cancer cells(DLD-1 and HCT116)and control cell(HcoEpic)were treated with different concentrations of zeylenone(0,2.5,5,10 and 20 μmol/L)for 48 h.Cell viability of DLD-1,HCT116 and HcoEpic were determined by CCK-8 kit.TUNEL staining was used to evaluate cell apoptosis.Caspase-3 activity and LDH level were measured by ELISA.Relative levels of M1 and M2 polarization markers,p-PI3K/PI3K and p-AKT/AKT were detected by Western blot and qRT-PCR.DLD-1 cells were subcuta-neously injected into the armpit of nude mice to establish a mouse xenograft tumor model.Intraperitoneal dose of zeylenone given to nude mice was 30 mg/kg,administered once every two days.After 2 weeks,the effect of zeylenone on growth of colorectal cancer tumors was detected.Results:Zeylenone inhibited cell activity and promoted cell apoptosis in colorectal cancer cells.Additionally,zeylenone stimulated M1-polarization and inhibited M2-polarization of tumor-associated macrophages(P<0.05).PI3K/AKT signaling pathway was inhibited by zeylenone in colorectal cancer cells(P<0.05).PI3K/AKT signaling pathway activator(740 Y-P)attenuated the effect of zeylenone on colorectal cancer cells.In mouse xenograft model,zeylenone inhibited the growth of colorectal cancer tumors(P<0.05).Conclusion:Zeylenone can inhibit colorectal cancer cell activity,promote colorectal cancer cell apoptosis,and promote M1 po-larization of tumor-associated macrophages by regulating PI3K/AKT signaling pathway,ultimately inhibiting the progression of colorec-tal cancer.

Objective: This study was to investigate the effects of MEHP on isolated rat heart and explore its mechanism. Methods: The experiments were performed with Langendorff-perfused rat heart with a Langendorff apparatus. 35 SD rats were used in the experiment and there were 5 rats per group. MEHP at doses of 3.125, 6.250, 12.500 and 25.000 μmol/L were given to the hearts for 25 minutes. Effects of NAC at concentration of 5 mmol/L were evaluated by co-treatment with 12.500 or 25.000 μmol/L MEHP. Data was collected per 5 minutes for 25 minutes. The heart rate, LVDP, LVEDP, dp/dtmax, and dp/dtmin were measured and analyzed using a PL3508 Data Acquisition and Analysis System. 200 waves at least were required each time. LDH contents in heart lavage fluid were determined by photometric assays using the automated biochemical analyzer. A section of the heart tissue was used for histopathological examination. DCFH-DA method was used to detect the levels of reactive oxygen species in different groups of heart tissues. Results: There was a concentration dependent decrease of heart rate (P<0.05) . At concentrations of 6.250, 12.500 and 25.000 μmol/L, MEHP significantly decreased the LVDP, dp/dtmax and dp/dtmin (P<0.05) , and this decrease is more pronounced with perfusion time. As the MEHP was given up to 6.250, 12.500 and 25.000 μmol/L, a statistical significance was found in the increase of LVEDP (P<0.05) . For dp/dtmin, a significant increase was observed at the concentration of 3.125 μmol/L when perfused with 10 and 15 min (P<0.05) , but this increase disappeared over time. LDH in cardiac perfusate increased as the MEHP given a higher concentration (P<0.05) . Compared with the control group, Histopathological analysis showed edema of myocardial tissue and cells, and inflammatory cells infiltration and myocardial cells necrosis were obvious in the MEHP perfusion groups. Myocardial ROS levels of the four MEHP treatment groups were all significantly higher than that of control group (P<0.05) . These heart damage induced by MEHP could be attenuated by NAC in different degrees. Conclusion MEHP can induce damage to myocardial tissue of isolated rat heart and one possible mechanism is the oxidative stress

Objective To explore the relationship between plasma microRNA 126 ( miR-126 ) level and coronary collateral circulation ( CCC) formation and to determine whether the miR-126 in plasma could serve as a blood-based biomarker for CCC in patients with severely narrowed coronary arteries ( CAD ).Methods In this prospective study , a total of 120 consecutive CAD patients with ≥95% stenosis in one epicardial coronary artery were enrolled.Thirty healthy people served as normal control.They were divided into two groups according to Rentrop grades:patients with grade 2 and 3 collateral development ( good CCC group, n=64) and patients with grade 0 and 1 collateral development (poor CCC group, n=56).Plasma miR-126 was measured by RT-PCR and serum VEGF was evaluated by ELISA method.Results Fasting plasma glucose ( FPG) was significantly lower in patients with good CCC than in patients with poor CCC ((5.99 ±1.48) mmol/L vs.(6.40 ±2.50) mmol/L).Plasma miR-126 levels and VEGF levels were significantly lower in CAD patients than in healthy people (0.04 ±0.01 vs.0.07 ±0.02, P=0.023 and (2 110 ±455) ng/L vs.(2 574 ±450) ng/L, P=0.011, respectively).miR-126 and VEGF levels were significantly higher in good CCC group than in poor CCC group ( miR-126:0.06 ±0.02 vs.0.03 ±0.01, P=0.021;VEGF:(2 549 ±614) ng/L vs.(1 759 ±452) ng/L, P=0.008).In CAD patients with good CCC, the miR-126 level was positively correlated to the VEGF expression (r =0.712,P=0.005) while there was no correlation between miR-126 level VEGF in CAD patients with poor CCC ( r =0.342, P =0.483).Multivariate analysis revealed that plasma miR-126 ( OR=2.145,95%CI 1.691 -2.988, P =0.001) and VEGF ( OR =1.279, 95%CI 1.068 -2.295, P =0.013 ) were independent predictors of collateral formation in patients with severely narrowed coronary arteries.In CAD patients , the area under the miR-126 ROC curve is 0.951 ( P=0.002).Conclusion Plasma miR-126 level is positively correlated to the CCC formation and is an independent predictor of CCC development in patients with severely narrowed coronary arteries , suggesting that plasma miR-126 might be a useful new , stable blood biomarker for predicting CCC formation in patients with severely narrowed coronary arteries.

Objective:To explore the rapid evaluation of the early pathogen of severe Chlamydophila psittaci pneumonia by bedside diagnostic bronchoscopy, so as to start effective anti-infection treatment before the results of macrogenome next generation sequencing (mNGS) test. Methods:The clinical data of three patients with severe Chlamydophila psittaci pneumonia who were successfully treated in the First Affiliated Hospital of Xinjiang Medical University, the First People's Hospital of Aksu District, and the First Division Hospital of Xinjiang Production and Construction Corps from October 2020 to June 2021 were retrospectively analyzed, including the rapid assessment of early pathogens by bedside diagnostic bronchoscopy and the use of antibiotics to start anti-infection treatment. These patients were successfully treated. Results:The three patients were male, aged 63, 45 and 58 years old, respectively. Before the onset of the penumonia, they had a clear medical history of bird exposure. The clinical manifestations mainly included fever, dry cough, shortness of breath and dyspnea. One case had abdominal pain and lethargy. The results of laboratory examination indicated that the peripheral blood white blood cell count (WBC) of two patients were high [(10.2-11.9)×10 9/L], the percentage of neutrophils increased (85.2%-94.6%) and the percentage of lymphocytes decreased (3.2%-7.7%) in all 3 patients after admission to hospital and entering into intensive care unit (ICU). The procalcitonin (PCT) of 3 patients increased after admission, and still increased when entering ICU (0.3-4.8 ng/L), so did C-reactive protein (CRP, 58.0-162.0 mg/L) and erythrocyte sedimentation rate (ESR, 36.0-90.0 mm/1 h). After admission, serum alanine transaminase (ALT) increased in 2 cases (136.7 U/L, 220.5 U/L), so did aspartate transaminase (AST) in 2 cases (249.6 U/L, 164.2 U/L). ALT (162.2-267.9 U/L) and AST (189.8-223.2 U/L) increased in 3 patients when they entered ICU. The level of serum creatinine (SCr) of 3 patients were normal after admission and entering ICU. The chest computed tomography (CT) findings of 3 patients were acute interstitial pneumonia, bronchopneumonia and lung consolidation, of which 2 cases were accompanied by a small amount of pleural effusion, and 1 case was accompanied by more regular small air sacs. Multiple lung lobes were involved, but mainly one lung lobe. The oxygenation index (PaO 2/FiO 2) of the 3 patients admitting to ICU were 100.0, 57.5 and 105.4 mmHg (1 mmHg ≈ 0.133 kPa), respectively, which met with the diagnostic criteria of moderate and severe acute respiratory distress syndrome (ARDS). All three patients received endotracheal intubation and mechanical ventilation. Under the bedside bronchoscope, the bronchial mucosa of 3 patients were obviously congested and edematous, without purulent secretion, and there was 1 case with mucosal hemorrhage. Three patients underwent bedside diagnostic bronchoscopy, and the evaluation result of the pathogen was that it might be atypical pathogen infection, so they were given moxifloxacin, cisromet and doxycycline intravenously, respectively, and combined with carbapenem antibiotics intravenously. After 3 days, the detection results of mNGS in bronchoalveolar lavage fluid (BALF) showed that only Chlamydia psittaci was infected. At this time, the condition was significantly improved, and PaO 2/FiO 2 was significantly increased. Therefore, the antibiotic treatment scheme remained unchanged, and mNGS only served to verify the initial diagnosis. Two patients were extubated on the 7th and 12th day of admission to the ICU, respectively, while one patient was extubated on the 16th day of admission to the ICU due to nosocomial infection. All 3 patients were transferred to the respiratory ward after the condition was stable. Conclusion:The bedside diagnostic bronchoscopy based on clinical characteristics is conducive to not only the rapid assessment of the early pathogens of severe Chlamydia psittaci pneumonia, but also effective anti-infection treatment before the returning of mNGS test results, which can make up for the lag and uncertainty of the mNGS test results.

ObjectiveTo investigate the protective effect of Geju Hugan tablets on the liver of mice with alcohol-induced liver injury， and explore the underlying mechanism based on nuclear factor-κB p65 （NF-κB p65） and B-cell lymphoma-2 （Bcl-2）/Bcl-2-associated X protein （Bax） signaling pathways. MethodAccording to the body weight， 60 SPF-grade male ICR mice were randomized into normal， model， Compound Yiganling tablets （0.16 g·kg^-1）， and low-， medium-， and high-dose （0.2， 0.4， 0.8 g·kg^-1， respectively） Geju Hugan tablets groups. The drugs were administrated at the corresponding doses by gavage， and the normal and model groups with equal volume of pure water once a day for 28 consecutive days. On day 29， the mice in other groups except the normal group were administrated with liquor （53% Vol） by gavage twice a day at the doses of 20， 10 mL·kg^-1 and with the interval of 6 h. Samples were harvested on day 30. The histopathological changes in the liver were observed by hematoxylin-eosin （HE） staining， and the ultrastructural changes in hepatocytes were observed by transmission electron microscopy. The enzyme-linked immunosorbent assay was employed to measure the levels of malonaldehyde （MDA）， reduced glutathione （GSH）， and triglycerides （TG） in the liver tissue and the levels of alanine aminotransferase （ALT） and aspartate aminotransferase （AST） in the serum. Western blotting was employed to determine the protein levels of NF-κB p65， phosphorylated p-inhibitor kappa B alpha （p-IκBα）， Bcl-2， and Bax in the liver tissue. ResultCompared with the normal group， the model group showed increases in the ALT， AST， MDA， and TG levels， a decrease in the GSH level， and increases in the liver injury scores evaluated based on the HE， oil red O， and transmission electron microscopy （P<0.01）. Moreover， the model group showed up-regulated expression of NF-κB， p-IκBα， and Bax （P<0.05， P<0.01） and down-regulated expression of Bcl-2 （P<0.05） in the liver tissue. Compared with the model group， Geju Hugan tablets of all the doses lowered the ALT， AST， MDA， and TG levels and elevated the GSH level （P<0.01）. The liver injury scores assessed based on HE staining and transmission electron microscopy in the medium- and high-dose Geju Hugan tablets groups were lower than those in the model group （P<0.01）. Compared with the model group， medium- and high-dose Geju Hugan tablets down-regulated the protein levels of NF-κB， p-IκBα， and Bax （P<0.01） and all doses of Geju Hugan tablets up-regulated the protein level of Bcl-2 （P<0.01）. ConclusionGeju Hugan tablets protect mice from alcohol-induced liver injury by down-regulating NF-κB signaling pathway to alleviate inflammation in the liver tissue and down-regulating the expression of Bax and up-regulating the expression of Bcl-2 to inhibit hepatocyte apoptosis.

OBJECTIVETo explore the relationship between plasma microRNA126 (miR-126) level and coronary collateral circulation (CCC) formation and to determine whether the miR-126 in plasma could serve as a blood-based biomarker for CCC in patients with severely narrowed coronary arteries (CAD).

METHODSIn this prospective study, a total of 120 consecutive CAD patients with ≥ 95% stenosis in one epicardial coronary artery were enrolled. Thirty healthy people served as normal control. They were divided into two groups according to Rentrop grades: patients with grade 2 and 3 collateral development (good CCC group, n = 64) and patients with grade 0 and 1 collateral development (poor CCC group, n = 56). Plasma miR-126 was measured by RT-PCR and serum VEGF was evaluated by ELISA method.

RESULTSFasting plasma glucose (FPG) was significantly lower in patients with good CCC than in patients with poor CCC ((5.99 ± 1.48) mmol/L vs. (6.40 ± 2.50) mmol/L). Plasma miR-126 levels and VEGF levels were significantly lower in CAD patients than in healthy people (0.04 ± 0.01 vs. 0.07 ± 0.02, P = 0.023 and (2 110 ± 455) ng/L vs. (2 574 ± 450) ng/L, P = 0.011, respectively). miR-126 and VEGF levels were significantly higher in good CCC group than in poor CCC group (miR-126: 0.06 ± 0.02 vs. 0.03 ± 0.01, P = 0.021;VEGF:(2 549 ± 614) ng/L vs. (1 759 ± 452) ng/L, P = 0.008) . In CAD patients with good CCC, the miR-126 level was positively correlated to the VEGF expression (r = 0.712, P = 0.005) while there was no correlation between miR-126 level VEGF in CAD patients with poor CCC (r = 0.342, P = 0.483) . Multivariate analysis revealed that plasma miR-126 (OR = 2.145, 95% CI 1.691-2.988, P = 0.001) and VEGF (OR = 1.279, 95% CI 1.068-2.295, P = 0.013) were independent predictors of collateral formation in patients with severely narrowed coronary arteries. In CAD patients, the area under the miR-126 ROC curve is 0.951 (P = 0.002).

CONCLUSIONPlasma miR-126 level is positively correlated to the CCC formation and is an independent predictor of CCC development in patients with severely narrowed coronary arteries, suggesting that plasma miR-126 might be a useful new, stable blood biomarker for predicting CCC formation in patients with severely narrowed coronary arteries.

Biomarkers ; Collateral Circulation ; Coronary Angiography ; Coronary Artery Disease ; blood ; Coronary Circulation ; Coronary Disease ; Heart ; Humans ; MicroRNAs ; blood ; Multivariate Analysis ; Plasma ; Prospective Studies ; ROC Curve

Age-dependent loss of skeletal muscle mass and function is a feature of sarcopenia, and increases the risk of many aging-related metabolic diseases. Here, we report phenotypic and single-nucleus transcriptomic analyses of non-human primate skeletal muscle aging. A higher transcriptional fluctuation was observed in myonuclei relative to other interstitial cell types, indicating a higher susceptibility of skeletal muscle fiber to aging. We found a downregulation of FOXO3 in aged primate skeletal muscle, and identified FOXO3 as a hub transcription factor maintaining skeletal muscle homeostasis. Through the establishment of a complementary experimental pipeline based on a human pluripotent stem cell-derived myotube model, we revealed that silence of FOXO3 accelerates human myotube senescence, whereas genetic activation of endogenous FOXO3 alleviates human myotube aging. Altogether, based on a combination of monkey skeletal muscle and human myotube aging research models, we unraveled the pivotal role of the FOXO3 in safeguarding primate skeletal muscle from aging, providing a comprehensive resource for the development of clinical diagnosis and targeted therapeutic interventions against human skeletal muscle aging and the onset of sarcopenia along with aging-related disorders.
Animals ; Humans ; Sarcopenia/metabolism* ; Forkhead Box Protein O3/metabolism* ; Muscle, Skeletal/metabolism* ; Aging/metabolism* ; Primates/metabolism*