Objective: To provide the mechanism-based pharmacotherapy of the Huatan Qushi formula (HTQS formula), for the health management and treatment of non-alcoholic fatty liver disease (NAFLD). Methods: A rat model of NAFLD was employed to examine the efficacy and safety of the HTQS formula. In vivo active components and potential mechanisms of the HTQS formula were identified using UPLC‒MS/MS combined with network pharmacology. The influence of the HTQS formula on the dominating proteins in PI3K/Akt pathway was validated in vivo using western blot. Finally, 16S rRNA sequencing of the gut microbiome was conducted followed by targeted metabolomics detecting fecal short-chain fatty acids (SCFAs) and bile acids to determine the impact of the HTQS formula on gut microbiota. Results: The HTQS formula reduced weight gain and hepatic steatosis in NAFLD rats and decreased serum total cholesterol (TC), triglycerides, blood glucose, and insulin resistance (IR) without causing liver or kidney injury. We detected 28 components using UPLC‒MS/MS and identified 439 shared targets between NAFLD and the HTQS formula. Primarily, we focused on the PI3K/Akt signaling pathway based on protein‒protein interaction network analysis. We validated that the HTQS formula inhibited liver steatosis and inflammation by increasing the phosphorylation levels of PI3K, AKT, P27, GSK3β in the PI3K/Akt signaling pathway. 16S rRNA sequencing revealed that the HTQS formula reduced the abundance of the genus Family_XIII_AD3011_group, which was positively correlated with IR and taurodeoxycholic acid. In addition, Lachnospiraceae_UCG_010 inversely correlated with TC and five bile acids, which could be essential to the therapeutic effect of the HTQS formula against NAFLD. Conclusions The HTQS formula proved to be an effective pharmacotherapy for NAFLD without causing liver or kidney injury. Multiple potent components of the HTQS formula could alleviate liver steatosis and lipid metabolism disorder by modulating the PI3K/Akt signaling pathway and restoring gut microbiota composition.

Objective:To explore the evaluation value of serum levels of positive pentameric protein 3 (PTX3) and creatine kinase isoenzyme MB (CK-MB) on volume load in patients with chronic decompensated heart failure (CDHF).Methods:A total of 300 CDHF patients who visited the Xingtai Central Hospital from July 2019 to July 2022 were selected and divided into a capacity overload group ( n=182) and a non capacity overload group ( n=118) based on their capacity balance level. Two clinical data sets were compared and analyzed. The receiver operating characteristic (ROC) curve was used to analyze the evaluation value of serum PTX3 and CK-MB levels on the volume load of CDHF patients. The clinical disease characteristics of the two groups of patients were analyzed using univariate analysis, and the influencing factors of volume load of CDHF patients were analyzed using logistic regression. A column chart model was constructed and validated. Results:The body mass index (BMI), waist circumference, systolic blood pressure, diastolic blood pressure, fasting blood glucose (FBG), glycosylated hemoglobin (HbA 1c), C-reactive protein (CRP), uric acid (UA), homeostasis model assessment of insulin resistance (HOMA-IR) of patients in the capacity overload group were higher than those in the non-capacity overload group, and the differences were statistically significant (all P<0.05). The PTX3, CK-MB, pulmonary capillary wedge pressure (PCWP), and CVP levels of patients in the capacity overload group were higher than those in the non-capacity overload group, while albumin, hemoglobin, and hematocrit were lower than those in the non-capacity overload group, and the differences were statistically significant (all P<0.05). The ROC curve showed that the area under the curve (AUC) of PTX3 and CK-MB for predicting capacity overload in CDHF patients are 0.795 and 0.718, with sensitivity of 86.2% and 83.7%, specificity of 65.4% and 68.6%, respectively, indicating high predictive accuracy; The AUC of the two joint predictions is 0.817, the sensitivity was 92.5%, and the specificity was 70.6%. The prediction accuracy was higher than PTX3 ( Z=3.812, P<0.05) and CK-MB ( Z=3.365, P<0.05). PTX3, CK-MB, albumin, hemoglobin, hematocrit, PCWP, and central venous pressure (CVP) were all influencing factors of volume load status in CDHF patients (all P<0.05). The column chart risk prediction model established based on these factors had high accuracy and strong applicability in clinical treatment. Conclusions:Serum PTX3 and CK-MB levels are influencing factors for volume overload in CDHF patients. A column chart model constructed in combination with indicators such as albumin, hemoglobin, hematocrit, PCWP, and CVP has high predictive value for the volume overload status of CDHF.

Objective:To evaluate the accuracy of the optical surface imaging system (OSI) using stereotactic radiosurgery (SRS) algorithm in single-center non-coplanar treatment of multiple brain metastases.Methods:Data of phantom and 15 patients with multiple brain metastases who underwent single-center non-coplanar radiotherapy in West China Hospital of Sichuan University from February to April 2022 were retrospectively analyzed. kV/MV and OSI imaging were used for imaging of the patients and phantoms under the same non-coplanar couch angle, respectively. The accuracy of OSI imaging of the phantoms and patients was evaluated using kV/MV imaging as reference image. The difference between the OSI and kV/MV systems is defined as accuracy, and the percentage of the absolute difference ≤1.00 mm in the translational direction or ≤0.50° in the rotational direction is defined as the threshold pass rate. Origin software was used to draw radar maps and Bland-Altman plots for statistical analysis.Results:When OSI images were used for the phantom imaging, the average differences in six-dimensional directions of lateral, long, vertical, rotational, roll and pitch were 0.03 mm, -0.09 mm, -0.27 mm, 0.04°, 0.17° and -0.19°, respectively. The maximum values were -2.20 mm, -2.30 mm, -1.20 mm, 0.60°, -1.00°, and -1.00°, respectively. When OSI system was utilized for the imaging of 15 patients, the average differences in six-dimensional directions were 0.44 mm, 0.16 mm, -0.20 mm, -0.11°, 0.10°, and -0.12°, respectively. The maximum values were -1.80 mm, 2.00 mm, 0.90 mm, -0.90°, -0.70°, and 0.80°, respectively. The translational errors mainly occurred in the lateral and long directions. The qualified rates of the threshold values of the phantoms and patients were 77% and 75% in the lateral direction, 82% and 89% in the long direction, respectively. In addition, 57% and 56% of patients met the threshold conditions of ±1.00 mm and ±0.50° in the six-dimensional directions, respectively.Conclusions:The OSI system using new SRS algorithm cannot meet the high accuracy requirements of single-center non-coplanar radiotherapy for multiple brain metastasis, especially in the lateral and long directions. It is not recommended for non-coplanar image guidance.

Background::Bladder cancer, characterized by a high potential of tumor recurrence, has high lifelong monitoring and treatment costs. To date, tumor cells with intrinsic softness have been identified to function as cancer stem cells in several cancer types. Nonetheless, the existence of soft tumor cells in bladder tumors remains elusive. Thus, our study aimed to develop a microbarrier microfluidic chip to efficiently isolate deformable tumor cells from distinct types of bladder cancer cells.Methods::The stiffness of bladder cancer cells was determined by atomic force microscopy (AFM). The modified microfluidic chip was utilized to separate soft cells, and the 3D Matrigel culture system was to maintain the softness of tumor cells. Expression patterns of integrin β8 (ITGB8), protein kinase B (AKT), and mammalian target of rapamycin (mTOR) were determined by Western blotting. Double immunostaining was conducted to examine the interaction between F-actin and tripartite motif containing 59 (TRIM59). The stem-cell-like characteristics of soft cells were explored by colony formation assay and in vivo studies upon xenografted tumor models. Results::Using our newly designed microfluidic approach, we identified a small fraction of soft tumor cells in bladder cancer cells. More importantly, the existence of soft tumor cells was confirmed in clinical human bladder cancer specimens, in which the number of soft tumor cells was associated with tumor relapse. Furthermore, we demonstrated that the biomechanical stimuli arising from 3D Matrigel activated the F-actin/ITGB8/TRIM59/AKT/mTOR/glycolysis pathways to enhance the softness and tumorigenic capacity of tumor cells. Simultaneously, we detected a remarkable up-regulation in ITGB8, TRIM59, and phospho-AKT in clinical bladder recurrent tumors compared with their non-recurrent counterparts.Conclusions::The ITGB8/TRIM59/AKT/mTOR/glycolysis axis plays a crucial role in modulating tumor softness and stemness. Meanwhile, the soft tumor cells become more sensitive to chemotherapy after stiffening, that offers new insights for hampering tumor progression and recurrence.

Bladder cancer remains the 10th most common cancer worldwide. In recent years, metformin has been found to have potential anti-bladder cancer activ-ity while high concentration of IC50 at millimolar level is needed, which could not be reached by regular oral administration route. Thus, higher efficient agent is urgently demanded for clinically treating bladder cancer. Here, by conjugating artesunate to metformin, a novel artesunate-metformin dimer triazine derivative AM2 was designed and synthesized. The inhibitory effect of AM2 on bladder cancer cell line T24 and the mechanism underlying was determined. Anti-tumor activity of AM2 was assessed by MTT, cloning formation and wound healing assays. Decreasing effect of AM2 on lipogenesis was determined by oil red O staining. The protein expressions of Clusterin, SREBP1 and FASN in T24 cells were evaluated by Western blotting. The results show that AM2 significantly inhibited cell proliferation and migration at micromolar level, much higher than parental metformin. AM2 reduced lipogenesis and down-regulated the expressions of Clusterin, SREBP1 and FASN. These results suggest that AM2 inhibits the growth of bladder cancer cells T24 by inhibiting cellular lipogenesis associated with the Clusterin/SREBP1/FASN signaling pathway.

Functionalized liposomes can improve the in vivo process of drugs to achieve high-efficiency delivery by enhancing drug absorption， changing drug distribution and reducing the elimination， which is one of the hotspots in nanomedicine research with broad application prospects. However， the drug information published by official websites of National Medical Products Administration （NMPA）， the United States Food and Drug Administration （FDA）， and the European Medicines Agency （EMA） shows that there are few liposomal products on the market， and the domestic varieties are mainly generic drugs. Excepting for polyethylene glycolized （PEGylated） liposomes， no other functionalized liposomes have been approved for marketing， which indicates that the clinical translation of functionalized liposomes remains at a low level. Therefore， the relevant reports of functionalized liposomes in recent years were reviewed in this paper， their application advantages and main challenges in preparation research and development were discussed based on the in vivo process， and their low clinical translation mainly because of the insufficient clinical thinking， safety and efficacy of functional materials， inaccurate in vitro and in vivo analysis methods and difficulty in scaling up production. Meanwhile， the possible strategies such as introducing the concept of clinical multi-function to improve clinical acuity， focusing on examining the modification density of functional materials and the interaction between the modified materials， evaluating the drug delivery performance of functionalized liposomes from multiple perspectives and scenarios， and conducting cost and convenience-oriented formulation composition and preparation process optimization were proposed in order to provide a reference for the development of functionalized liposomes and other carrier-based nanomedicines.

Male ; Humans ; Aged ; Sleep Duration ; Aging ; Testosterone ; China ; Sleep

Objective:To explore the chain mediating effect of core self-evaluation and social withdrawal on the relationship between peer relationship and depression symptoms of adolescents.Methods:From September 2020 to October 2020, a sample of 1 936 students from grade 4 to grade 9 of different schools completed a cross-section questionnaire survey including the inventory of parent and peer attachment, core self-evaluation scale, child social preference scale-R and depression self-rating scale for children.SPSS 21.0 and SPSS PROCESS macro program were used for data statistics.The statistical methods included analysis of variance, correlation analysis and intermediary effect test.Results:(1) Peer relationship (94.78±17.27) was positively correlated with core self-evaluation (34.14±7.52) ( r=0.50, P<0.01), and negatively correlated with depression (12.21±6.02) and social withdrawal (32.34±11.45) ( r=-0.55, -0.58, both P<0.01). Core self-evaluation was negatively associated with social withdrawal and depression symptoms ( r=-0.48, -0.67, both P<0.01), while social withdrawal and depression was positively correlated ( r=0.54, P<0.01). (2) Peer relationship had a significant direct effect on depression symptoms (the effect value=-0.205, P<0.01). Core self-evaluation and social withdrawal respectively separate mediated the effect of peer relationship on depression symptoms (the effect value=-0.231, -0.088, 95% CI=-0.261--0.202, -0.110--0.068), while the chain mediating effect of the two was significant, and the effect value was -0.025(95% CI=-0.033--0.019). Conclusion:Good peer relationship can lead to higher core self-evaluation and less social withdrawal behaviors in adolescents that serves as buffer from depression.

Objective:To observe the effect of repeated, bilateral administration of high-frequency transcranial magnetic stimulation (rTMS) in treating post-stroke dysphagia.Methods:Forty-five persons with post-stroke dysphagia were randomly divided into a bilateral group ( n=14 after one dropout), an affected group ( n=15) and a healthy group ( n=15). All received 30 minutes of conventional swallowing rehabilitation training 5 times a week for 2 weeks from a speech therapist. Those in the affected group also received 5Hz rTMS applied to the motor cortex controlling the suprachyoid muscle group. The bilateral group received the same stimulation bilaterally with the same duration and treatment course. Videofluoroscopy was used to assess their swallowing before and after the 2 weeks of treatment. It was rated using the penetration-aspiration scale (PAS) and the functional swallowing disorder scale (FDS). Surface electromyography was employed to evaluate suprachyoid muscle function. Cortical excitability was assessed by measuring the resting motor threshold (RMT) of the unaffected hemisphere. Results:After the treatment, the average PAS, FDS and muscle function values had improved significantly for all three groups, but significant RMT differences were observed only between the bilateral and the unaffected group. Significant differences in the average FDS and PAS scores were observed after the treatment, as well as significant changes in FDS and muscle function between the affected group and the other two groups. The average FDS scores before and after treatment were significantly different between the unaffected and bilateral group, with the former scoring significantly better than the latter. But no significant differences in the average PAS scores were observed after the treatment.Conclusions:5Hz rTMS of either the unaffected or affected cerebral cortex (or bilateral) can effectively improve the swallowing function of persons with post-stroke dysphagia. Bilateral stimulation has the greatest therapeutic effect, followed by stimulation of the unaffected cerebral cortex.

Objective: To investigate the differential expression of serum-and-glucocorticoid-inducible-kinase-2 (SGK2) in hepatocellular carcinoma (HCC) and normal liver tissues and the related mechanism mediating signal transduction of GSK-3 β / β catenin in HCC cells. Methods: Twenty pairs of matched HCC and normal tissues were collected and the situation of expression of SGK2 mRNA was detected by real-time fluorescence quantitative PCR. Western blot was used to detect the levels of SGK2 protein in human HCC cell lines (Huh-7, SMMC-7721) and normal human liver cell line (L02). SGK2 siRNA was used to transfect human HCC cell lines (SMMC-7721 and Huh-7), and then the protein expression levels of GSK-3 β/ β - catenin was successfully detected with the above-mentioned transfected cell line by western blot. Measurement data were expressed as mean ± standard deviation (±s), and the Student t -test was used as the statistical method. Results: SGK2 mRNA expression was up-regulated in all 20 HCC samples than that of the expression of matched normal liver tissues. SGK2 protein levels were significantly higher in Huh-7 and SMMC-7721 than normal human liver cell lines (P < 0.01). The downregulation of SGK2 expression in human HCC cell lines (SMMC-7721 and Huh-7) had inhibited the expression of unphosphorylated GSK-3 β. In addition, the downregulation of SGK2 expression in HCC cell lines had decreased the dephosphorylation of β - catenin to prevent degradation of the β - catenin proteasome. Conclusion SGK2 is overexpressed in HCC and mediates GSK-3β/β- catenin signaling in HCC cells.