Objective To explore the regulatory effect of miR-181a on PTEN-induced kinase 1 (PINK1)/Parkin-related genes (Parkin) pathway, and on mitochondrial autophagy of osteoclasts in osteoporosis (OP) rats. Methods Twenty healthy female SD rats were randomly divided into osteoporosis (OP) model group and control group (10 each). Rats in OP model group were employed to prepare OP model. Osteoclasts were extracted from OP rats, and set them as: OP group (not transfected), si-miR-181a group (transfected with si-miR-181a vector plasmid), si-NC group (transfected with negative control plasmid), ad-miR-181a group (transfected with ad-miR-181a vector plasmid), and ad-NC group (transfected with negative ad-NC control plasmid),and the osteoclasts of rats in control group were cultured normally and set as normal control group. The expression of miR-181a was detected by RT-PCR, and MTT and flow cytometry were performed to detect the survival and apoptosis of osteoclasts, the mitochondria autophagy was observed with transmission electron microscope, the expression of Parkin in mitochondria was detected by immunofluorescence co-localization, Western blotting was performed to detect the expression of PINK1/Parkin and autophagy,as well as apoptosis related proteins. Knockdown of miR-181a in osteoclasts of OP rats to down regulate the expression of Parkin and verify the reversal effect of Parkin on miR-181a. Double Luciferase Report experiment verified the targeted regulation between miR-181a and Parkin. Results Compared with normal control group, miR-181a (1.59±0.15 vs. 1.02±0.11), Parkin⁺TOMM2⁺(2.02±0.20 vs. 0.13±0.10), Parkin (1.83±0.18 vs. 1.13±0.10) in osteoclasts of OP rats, and PINK1 (1.93±0.19 vs. 1.03±0.10)expression and survival rate (157.06%±12.32% vs. 100.09%±0.05%), autophagy marker protein-LC3-Ⅱ/Ⅰ ratio (1.89±0.18 vs. 1.15±0.10) increased (P<0.05). Compared with OP group, after up-regulating the expression of miR-181a in osteoclasts of OP rats, the cell survival rate (222.96%±22.15%) increased, Parkin⁺TOMM2⁺ (1.01±0.11), LC3-Ⅱ/Ⅰ ratio (1.36±0.12) and apoptosis rate (3.28%±0.35%) decreased (P<0.05). While after down-regulating the expression of miR-181a in osteoclasts of OP rats, the cell survival rate (106.96%±10.15%) decreased, Parkin⁺TOMM2⁺ (2.97±0.29), LC3-Ⅱ/Ⅰ ratio (2.47±0.24) and apoptosis rate (19.71%±1.83%) increased (P<0.05). The dual luciferase reporter test showed that Parkin is the target gene of miR-181a. Down-regulating Parkin expression can reverse the effects of miR-181a low expression in promoting mitochondrial autophagy and inhibiting cell survival (P<0.05). Conclusion Up-regulation of miR-181a expression in OP rat's osteoclasts can target down-regulation of Parkin expression, inhibit activation of mitochondrial autophagy, and promote the survival of osteoclasts.

Functional metabolomics is an extension of metabolomics, through detecting changes in endogenous metabolites that are stimulated and disturbed by a particular organism, after obtaining the biomarker, experiments were conducted to verify the functions of the biomarker and the associated enzymes, genes and proteins. The molecular mechanism of pathophysiology can be revealed systematically from the perspective of "metabolite-enzyme-gene-protein", solving the upstream and downstream biological mechanisms of metabolite association. This article reviews and analyzes the current common methods and research strategies of functional metabolomics, the exploration of disease targets and pathogenesis, and the analysis of drug action mechanisms in the field of biomedicine. At the same time, the extended research on the combination of functional metabolomics with network pharmacology and reprogramming metabolomics is summarized, and finally the application prospects of functional metabolomics are analyzed and prospected in the field of biomedicine.

Objective To investigate the role and mechanism of myotubularin-related protein 7 (Mtmr7) in proliferation and migration of mouse vascular smooth muscle cells (VSMCs). Methods The mouse aortic smooth muscle cell line (MOVAS)was cultured. 30 ng/ml of platelet-derived growth factor-BB (PDGF-BB) was used to induce proliferation and migration of VSMCs in vitro. The changes of mRNA and protein expression levels after PDGF-BB intervention in Mtmr7 was assessed by qRT-PCR and Western blotting. To explore the role of Mtmr7 in proliferation and migration of mouse VSMCs, the adenovirus carrying Mtmr7(Ad-Mtmr7) was used to infect VSMCs for overexpression of Mtmr7. MOVAS was divided into control group, Ad-Mtmr7 group,PDGF-BB group and Ad-Mtmr7+PDGF-BB group. The proliferation capacity of VSMCs was analyzed by Ki-67 immunofluorescence staining and cell counting kit-8 (CCK-8) assay. The migration capacity was assessed by scratch assay. The downstream target protein levels of mammalian rapamycin target protein complex 1 (mTORC1) were determined by Western blotting. Insulin (5 mg/L)was used to restore the activity of mTORC1. MOVAS were divided into PDGF-BB group, Ad-Mtmr7+PDGF-BB group and Ad-Mtmr7+PDGF-BB+insulin group. The proliferation, migration and protein levels were measured by methods as the same mentioned above. A model of carotid endothelial injury was established. At 28 days after operation, the protein level of Mtmr7 was determined by Western blotting. The mice were randomly divided into 4 groups (10 each): sham group, sham+Ad-Mtmr7 group,carotid endothelial injury group and carotid injury+Ad-Mtmr7 group. To overexpress Mtmr7, Ad-Mtmr7 (5×10¹⁰ pfu/ml) was injected into the carotid artery immediately after operation and then partly incubated for 30 min. At 7th, 14th and 21st day after operation, the mice were injected the adenovirus via tail vein. Twenty-eight days after modeling, the morphology of carotid artery and the degree of intimal hyperplasia were analyzed by HE staining. Results Compared with control group, the mRNA and protein levels of Mtmr7 were obviously reduced (P<0.001 and P<0.05), the rate of Ki-67 positive cells and the relative number of VSMCs increased (P<0.01 and P<0.001), the rate of wound healing and the protein expression levels of p-S6^Ser235/236 and p-4EBP1^Thr37/46 increased (P<0.001 and P<0.05) in PDGF-BB group. Compared with the PDGF-BB group, the rate of Ki-67 positive cells, the relative number of VSMCs (P<0.01 or P<0.001), the rate of wound healing (P<0.001) and the protein levels of p-S6^Ser235/236 and p-4EBP1^Thr37/46 (P<0.05) were decreased in Ad-Mtmr7+PDGF-BB group. Compared with the Ad-Mtmr7+PDGF-BB group, the protein levels of p-S6^Ser235/236 and p-4EBP1^Thr37/46 (P<0.01), the rate of Ki-67 positive cells, the relative number of VSMCs (P<0.01 or P<0.001) and the rate of wound healing (P<0.01) were increased in Ad-Mtmr7+PDGF-BB+insulin group. Compared with the sham group, the protein expression level of Mtmr7 decreased significantly (P<0.01) and the ratio of intima/media area increased (P<0.001)in carotid endothelial injury group. Compared with the carotid endothelial injury group, after overexpression of Mtmr7, the ratio of intima/media area decreased significantly (P<0.01) in carotid endothelial injury+Ad-Mtmr7 group. Conclusion Overexpression of Mtmr7 may inhibit the proliferation and migration of VSMCs induced by PDGF-BB in mice, alleviating intimal hyperplasia after vascular injury, which is closely related to the mTORC1 activity reduced by Mtmr7.

Objective To investigate the effects and mechanism of short chain fatty acids (SCFAs) on lipopolysaccharide(LPS) induced acute respiratory distress syndrome (ARDS) in rats. Methods Twenty male SD rats were randomly divided into control group, model group, low-dose SCFAs group and high-dose SCFAs group (5 each). Rats in control and model groups were given normal saline, in low-dose SCFAs group were given sodium acetate 300 mg/kg, sodium propionate 100 mg/kg and sodium butyrate 100 mg/kg, and in high-dose SCFAs were given sodium acetate 600 mg/kg, sodium propionate 200 mg/kg and sodium butyrate 200 mg/kg by gavage for 7 days in advance. And then the rats in control group were perfused with normal saline, and in the other 3 groups were perfused with LPS (5 mg/kg) into trachea to establish ARDS model. The rats were sacrificed 12 hours after modeling, and the arterial oxygen partial pressure (PaO₂) and lung wet/dry weight ratio (W/D) were measured, and HE staining was performed to observe the pathological changes of lung tissues. The concentrations of tumour necrosis factor (TNF)-α, interleukin(IL)-6 and IL-10 in serum and bronchoalveolar lavage fluid (BALF) were detected by ELISA. Western blotting was used to detect c-Jun N-terminal kinase (JNK), p-JNK, extracellular regulated protein kinases (ERK), p-ERK, p38 mitogen-activated protein kinase(MAPK), p-p38 MAPK, nuclear factor kappa-B (NF-κB) p65 and p-NF-κB p65 protein expression in lung tissues. The expression of tight junction protein ZO-1 and Occludin in colonic tissue were detected by immunohistochemistry. Results Compared with control group, PaO₂ decreased, lung W/D and lung pathological injury score increased, the concentration of TNF-α and IL-6 in serum and BALF significantly increased, and the concentration of IL-10 increased, the relative protein expressions of p-JNK/JNK,p-ERK/ERK, p-p38 MAPK/p38 MAPK and p-NF-κB p65/NF-κB p65 in lung tissues significantly increased, while the relative expression of ZO-1 and Occludin protein significantly decreased in colonic tissues in model group (P<0.05). Compared with model group, PaO₂ increased, lung W/D and lung pathological injury score decreased, and TNF-α, IL-6 in serum and BALF decreased,the concentration of IL-10 increased further, while the relative expressions of p-JNK/JNK, p-ERK/ERK, p-p38 MAPK/p38 MAPK and p-NF-κB p65/NF-κB p65 protein in lung tissue decreased, and the relative expression of tight junction protein ZO-1 and Occludin in colon tissue increased in low and high dose SCFAs groups (P<0.05). Compared with low-dose SCFAs group, the indexes mentioned above in the high-dose SCFAs group improved more significantly (P<0.05). Conclusion SCFAs could regulate immunity and inflammation, and effectively improved LPS-induced ARDS in rats, the mechanism might be related to enhancement of intestinal barrier and inhibition of MAPK and NF-κB signaling pathway activation, thereby reducing the release of inflammatory factors and increasing the production of anti-inflammatory factors.

Acute hepatitis of unknown etiology in children has been reported in many countries all over the world since April 2022. As of May 27,there have been at least 650 cases of such patients reported by World Health Organization in 33 countries. The etiology of the hepatitis is still unknown.Adenovirus has been detected in samples of some cases, but it is also suspected that the immune response triggered by SARS-CoV-2 infection maybe contribute to the mechanism of the disease. In this paper, we overview the research and reports about the acute hepatitis of unknown etiology in children about its epidemiology,clinical features and possible etiology, then for the aim to make our country doing well in the monitoring and controlling of the acute hepatitis of unknown etiology in children. Here, we present some points of view from the perspective of infectious diseases and liver diseases in children.

Objective To establish an improved method for estimating the mean blood glucose by glycosylated hemoglobin(HbA_1c) in diabetic patients with renal insufficiency. Methods The clinical data were retrospectively analyzed of 329 patients with type 2 diabetes mellitus who were hospitalized in the Department of Hypertension and Endocrinology of Daping Hospital of Army Medical Center of PLA from January 2018 to December 2021. All patients were divided into control group [estimated glomerular fraction rate, eGFR≥60 ml/(min.1.73 m²), n=165] and renal insufficiency group [eGFR<60 ml/(min.1.73 m²), n=164]based on their eGFR levels. The basic demographic data, oral glucose tolerance test (OGTT) 2 hours post plasma glucose, and other laboratory test results of the patients were collected. The HbA_1c and OGTT models were adopted separately to estimate the 24-hour average blood glucose of the enrolled patients, and the gap glucose level of the two methods was calculated. The influencing factors of estimating blood glucose deviation with HbA_1c in patients with renal insufficiency were analyzed, and a correction model of estimating average blood glucose with HbA_1c was established. A total of 29 patients with type 2 diabetes mellitus and renal insufficiency hospitalized in the Department of Hypertension and Endocrinology of Daping Hospital of Army Medical Center of PLA from January to March 2022 were collected for external verification. Results Compared with control group, diabetic patients with renal insufficiency were relatively older (P=0.001), with longer course of diabetes (P<0.001), and had lower HbA_1c (P=0.034),eGFR (P<0.001), Hb (P<0.001), and ALB (P<0.001) levels. The mean blood glucose of the two groups was estimated by HbA_1c. The gap of estimated blood glucose was significantly higher in renal insufficiency group than in control group [(0.88±1.64) mmol/L vs.(–0.09±2.10) mmol/L, P<0.001], and was negatively correlated with the Hb level (r=–0.377, P<0.001), while with no relation with ALB level (P=0.551). A linear regression model was used to include HbA_1c, Hb, age, diabetes course, eGFR and ALB into the analysis,and to establish an estimation model for estimating the daily mean blood glucose of diabetic patients with renal insufficiency: mean blood glucose (mmol/L) =4.539+0.95×HbA_1c-0.016×Hb (adjusted R²=0.829, P<0.001). Using this model, the gap glucose level between the estimated mean blood glucose and the detected blood glucose were remarkably reduced in renal insufficiency group [(0.02±1.05) mmol/L vs. (0.88±1.64) mmol/L, P<0.001]. The goodness-of-fit index RNL=0.830 was obtained by external validation of 29 patients, indicating that the model obtained is highly feasible. Conclusions A correction model of estimating mean blood glucose by HbA_1c for diabetic patients with renal insufficiency has been successfully established. It is of great significance to further develop individualized HbA_1c control and improve the prognosis of diabetes patients.

Objective To investigate the predictive value of lactate (Lac) and complex model for the occurrence of prolonging mechanical ventilation (PMV) after off-pump coronary bypass grafting (OPCABG) based on dose-response analysis and decision curve analysis. Methods A retrospective analysis was conducted on 683 patients who underwent OPCABG from January to December 2019 in the Department of Cardiovascular Surgery of the General Hospital of the Northern Theater Command. These patients were divided into PMV group (n=107) and non-PMV group (n=576) based on whether duration of prolonged mechanical ventilation was longer than 24 h. The arterial blood lactate at 0 h and 6 h after admission to the ICU of cardiovascular surgery,baseline data and other clinical indicators of the patients were recorded. Preoperative, intraoperative and postoperative risk factors affecting PMV time were analyzed by univariate analysis, and a prediction model was established by indicators selected by logistic regression. Restricted cubic spline model, decision curve analysis (DCA) and receiver operating characteristic (ROC) curve were used to evaluate the predictive value of arterial blood lactate and complex model. Results There were no significant differences between PMV group and non-PMV group in gender, age, BMI, NYHA cardiac function classification, history of myocardial infarction, history of PCI, smoking history, hypertension, diabetes, hypercholesterolemia, >50% stenosis with of 3 coronary artery branches and left aortic stenosis >50%, number of blood vessel bridge, arterial blood lactate at 0 h after admission to ICU,preoperative serum creatinine, hemoglobin after operation, preoperative total bilirubin, preoperative direct bilirubin (P>0.05). There were statistically significant differences in the use of IABP, left ventricular ejection fraction (LVEF), pulmonary arterial pressure,preoperative red blood cell distribution width (RDW), arterial blood lactate at 6 h after admission to ICU, preoperative hemoglobin,preoperative hypersensitive CRP (hs-CRP), postoperative hs-CRP, preoperative troponin T (TNT) and preoperative amino-terminal pro-brain natriuretic peptide (NT-proBNP) (P<0.05). Multivariate logistic regression analysis showed that preoperative RDW, arterial blood lactate 6 h after admission to ICU, pulmonary arterial pressure, preoperative NT-proBNP were risk predictors of PMV, and use of IABP was protective predictor of PMV (P<0.05), OR of five factors were 1.242 (95%CI 1.001-1.539), 1.370 (95%CI 1.171-1.604), 1.043 (95%CI 1.002-1.087), 2.065 (95%CI 1.333-3.200), 0.146 (95%CI 0.071-0.301), respectively (P<0.05). The area under the receiver operating characteristic curve of arterial blood lactate 6 h after admission to ICU and complex model were 0.582 (95%CI 0.518-0.646), 0.727 (95%CI 0.674-0.781), respectively. Intensity of association between Lac 6 h and the development of PMV exhibited a non-linear dose response relationship (P<0.01). Decision curve analysis showed that compared with Lac 6 h, the complex model had a higher net benefit when the threshold probability was between 0.05 and 0.75. Conclusions Compared with Lac 6 h, complex model has a higher predictive value for the occurrence of PMV after OPCABG.

With the change of people's lifestyle and diet structure, hypertriglyceridemia (HTG) has gradually become an important factor causing or aggravating acute pancreatitis (AP). Some studies have pointed out that HTG has become the second major factor of AP following biliary diseases. The concept of hypertriglyceridemic-related acute pancreatitis (HTG-AP) has been put forward, and a large number of studies have been carried out on its mechanism. However, the mechanism of hypertriglyceridemia involved in the progression of AP is still unclear, leading to the lack of specific targets for treatment. This article reviews the pathogenesis of HTG-AP, including excessive free fatty acids (FFA)-induced oxidative stress, Ca²⁺ overload, endoplasmic reticulum stress, microcirculatory disturbance, etc.

Objective To explore the effect and mechanism of phosphatase and tensin homolog (PTEN) on renal interstitial fibrosis in diabetic nephropathy. Methods We culture NRK52E cells in vitro, cells were randomly assigned to three groups: normal glucose group (NG), high glucose group (HG) and mannitol group (MA), Western blotting was used to detect the protein levels of PTEN, LC3, P62, collagen-Ⅰ and Ⅲ in vitro. To detect the effect of PTEN on autophagy and collagen, the NRK52E were randomly assigned to three groups: NG group, HG group and HG+pPTEN group, the autophagy changes were observed by laser confocal microscopy, and the changes of collagen were observed by fluorescence microscopy. Results Western blotting in vivo showed that, compared with NG group, the expression levels of PTEN and LC3 were decreased, but of P62, collagen-Ⅰ and Ⅲ obviously increased in HG group (P<0.01), while no obvious change of the expression of PTEN and LC3, P62, collagen-Ⅰ and Ⅲ in MA group. The confocal microscope showed that, the number of red spots and yellow spots were decreased and significant autophagy inhibition in HG group, there were no significant autophagy inhibition in other two groups. The cellular immunofluorescence staining showed that, the red fluorescence of PTEN and protein expression were decreased, the collagen fluorescence staining and protein expression were increased in HG group, there were no significant change in other two groups. Conclusions In NRK52E cells under high glucose, PTEN expression reduced significantly,autophagy was inhibited, and then aggravate renal fibrosis. Overexpression of PTEN can restore autophagy and alleviate the progression of renal interstitial fibrosis.

Objective To investigate the effect of dexmedetomidine on perioperative anxiety of patients under general anesthesia in thoracic surgery. Methods One hundred and twenty patients undergoing elective surgery under general anesthesia in the Department of Thoracic Surgery, the Sixth Medical Center of Chinese PLA General Hospital, from June 2021 to September 2021, were enrolled and randomly divided into control group (no dexmedetomidine perioperative), D1 group (dexmedetomidine 0.6 μg/kg intraoperatively), and D2 group (dexmedetomidine 0.6 μg/kg intraoperative, dexmedetomidine 2.4 μg/kg postoperative analgesia pump), 40 patients in each group. Perioperative anesthetic dosage, anesthetic time, operation time, time from operation completion to extubation, time from extubation to out-room and time from postoperative to discharge were compared among the three groups. The patients' average arterial pressure (MAP) and heart rate (HR) at entry (T₀), after intubation (T₁), immediately after dexmedetomidine pump injection (T₂), before catheterization (T₃), at exit (T₄), and 24, 48, 72 h after operation were recorded.Self-rating anxiety scale (SAS), visual analogue scale-anxiety (VAS-a), visual analogue scale (VAS), and sedation score (Ramsay)were measured before and 24, 48, 72 h postoperatively. Results No significant differences were observed in operation time,anesthesia time, time from operation completion to extubation, time from extubation to out-room and anesthesia dosage among the three groups (P>0.05), but the time from postoperative to discharge was significantly shorter in D2 group than in control group[(7.13±3.83) d vs. (8.93±3.67) d, P=0.027]. Compared with control group, the MAP of patients in D1 group and D2 group decreased at T₃ (P<0.05). No significant differences were observed in MAP, HR and Ramsay scores among the three groups at 24,48 and 72 h postoperatively (P>0.05). The SAS scores of patients in the three groups decreased significantly at 48 and 72 h after operation, and the decrease was most obvious in D2 group (P<0.05); The VAS-a score was significantly lower in D2 group than in control group and D1 group at 24, 48 and 72 h after surgery (P<0.05). The VAS scores of patients in the three groups decreased gradually at 48 and 72 h after surgery than that at 24 h after surgery (P<0.05). Multivariate linear regression analysis showed that age, sex and length of preoperative hospital stay were significantly correlated with preoperative anxiety in thoracic surgery patients(P<0.05). Conclusion Intra- and post-operative application of dexmedetomidine can significantly improve perioperative anxiety,synergistic analgesia and shorten postoperative hospital stay in patients of thoracic surgery.