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 investigate the effect of respiratory training based on core stability training on feedforward control in pa-tients with chronic nonspecific low back pain(CNLBP). Methods A total of 60 patients with CNLBP in Jiaxing Second Hospital from January,2022 to March,2023 were ran-domly divided into control group(n=30)and experimental group(n=30).Both groups received health educa-tion,physical factor therapy and core stability training,while the experimental group received respiratory training in addition,for four weeks.Visual Analogue Scale(VAS)score,Japanese Orthopaedic Association low back pain(JOA)score and Oswestry Dysfunction Index(ODI)were compared between two groups before and after treat-ment,while surface electromyography was used to detect the root mean square(RMS)and integrated electromy-ography(iEMG)of transversus abdominis,multifidus and triceps(movement muscles),and the activation se-quence and relative activation time of transversus abdominis,multifidus and triceps were calculated. Results After treatment,the scores of VAS,JOA and ODI improved significantly in both groups(|t|>8.515,P<0.001),and the scores were better in the experimental group than in the control group(|t|>2.089,P<0.05).RMS and iEMG of transversus abdominis and multifidus improved significantly after treatment in both groups(|t|>18.831,P<0.001),and were significantly better in the experimental group(|t|>3.481,P<0.05).The transversus abdominis and multifidus in both groups were activated before the movement muscles,and the relative activation time of transversus abdominis and multifidus increased in negative(|t|>48.115,P<0.001),the experimental group being better(|t|>3.229,P<0.05). Conclusion Combination of core stability training and respiratory training is beneficial in reducing the pain of patients with CNLBP,reducing the lumbar dysfunction,improving the order of muscle activation,and strengthening feed-forward control.

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.

Objective:To develop and evaluate metal artifact removal systems (MARSs) based on deep learning to assess their effectiveness in removing artifacts caused by different thicknesses of metals in cone-beam CT (CBCT) images.Methods:A full-mouth standard model (60 mm×75 mm×110 mm) was three-dimensional (3D) printed using photosensitive resin. The model included a removable and replaceable target tooth position where cobalt-chromium alloy crowns with varying thicknesses were inserted to generate matched CBCT images. The artifacts resulting from cobalt-chromium alloys with different thicknesses were evaluated using the structural similarity index measure (SSIM) and peak signal-to-noise ratio (PSNR). CNN-MARS and U-net-MARS were developed using a convolutional neural network and U-net architecture, respectively. The effectiveness of both MARSs were assessed through visualization and by measuring SSIM and PSNR values. The SSIM and PSNR values were statistically analyzed using one-way analysis of variance (α=0.05).Results:Significant differences were observed in the range of artifacts produced by different thicknesses of cobalt-chromium alloys (all P<0.05), with 1 mm resulting in the least artifacts. The SSIM values for specimens with thicknesses of 1.0 mm, 1.5 mm, and 2.0 mm were 0.916±0.019, 0.873±0.010, and 0.833±0.010, respectively ( F=447.89, P<0.001). The corresponding PSNR values were 20.834±1.176, 17.002±0.427, and 14.673±0.429, respectively ( F=796.51, P<0.001). After applying CNN-MARS and U-net-MARS to artifact removal, the SSIM and PSNR values significantly increased for images with the same thickness of metal (both P<0.05). When using the CNN-MARS for artifact removal, the SSIM values for 1.0, 1.5 and 2.0 mm were 0.938±0.023, 0.930±0.029, and 0.928±0.020 ( F=2.22, P=0.112), while the PSNR values were 30.938±1.495, 30.578±2.154 and 30.553±2.355 ( F=0.54, P=0.585). When using the U-net-MARS for artifact removal, the SSIM values for 1.0, 1.5 and 2.0 mm were 0.930±0.024, 0.932±0.017 and 0.930±0.012 ( F=0.24, P=0.788), and the PSNR values were 30.291±0.934, 30.351±1.002 and 30.271±1.143 ( F=0.07, P=0.929). No significant differences were found in SSIM and PSNR values after artifact removal using CNN-MARS and U-net-MARS for different thicknesses of cobalt-chromium alloys (all P>0.05). Visualization demonstrated a high degree of similarity between the images before and after artifact removal using both MARSs. However, CNN-MARS displayed clearer metal edges and preserved more tissue details when compared with U-net-MARS. Conclusions:Both the CNN-MARS and U-net-MARS models developed in this study effectively remove the metal artifacts and enhance the image quality. CNN-MARS exhibited an advantage in restoring tissue structure information around the artifacts compared to U-net-MARS.

Objective:To develop and evaluate metal artifact removal systems (MARS) based on deep learning to assess their effectiveness in removing artifacts caused by different thicknesses of metals in cone-beam CT (CBCT) images.Methods:A full-mouth standard model (60 mm×75 mm×110 mm) was three-dimensional (3D) printed using photosensitive resin. The model included a removable and replaceable target tooth position where cobalt-chromium alloy crowns with varying thicknesses were inserted to generate matched CBCT images. The artifacts resulting from cobalt-chromium alloys with different thicknesses were evaluated using the structural similarity index measure (SSIM) and peak signal-to-noise ratio (PSNR). CNN-MARS and U-net-MARS were developed using a convolutional neural network and U-net architecture, respectively. The effectiveness of both MARSs were assessed through visualization and by measuring SSIM and PSNR values. The SSIM and PSNR values were statistically analyzed using one-way analysis of variance (α=0.05).Results:Significant differences were observed in the range of artifacts produced by different thicknesses of cobalt-chromium alloys (all P<0.05), with 1 mm resulting in the least artifacts. The SSIM values for specimens with thicknesses of 1.0, 1.5, and 2.0 mm were 0.916±0.019, 0.873±0.010, and 0.833±0.010, respectively ( F=447.89, P<0.001). The corresponding PSNR values were 20.834±1.176, 17.002±0.427, and 14.673±0.429, respectively ( F=796.51, P<0.001). After applying CNN-MARS and U-net-MARS to artifact removal, the SSIM and PSNR values significantly increased for images with the same thickness of metal (both P<0.05). When using the CNN-MARS for artifact removal, the SSIM values for 1.0, 1.5 and 2.0 mm were 0.938±0.023, 0.930±0.029, and 0.928±0.020 ( F=2.22, P=0.112), while the PSNR values were 30.938±1.495, 30.578±2.154 and 30.553±2.355 ( F=0.54, P=0.585). When using the U-net-MARS for artifact removal, the SSIM values for 1.0, 1.5 and 2.0 mm were 0.930±0.024, 0.932±0.017 and 0.930±0.012 ( F=0.24, P=0.788), and the PSNR values were 30.291±0.934, 30.351±1.002 and 30.271±1.143 ( F=0.07, P=0.929). No significant differences were found in SSIM and PSNR values after artifact removal using CNN-MARS and U-net-MARS for different thicknesses of cobalt-chromium alloys (all P>0.05). Visualization demonstrated a high degree of similarity between the images before and after artifact removal using both MARS. However, CNN-MARS displayed clearer metal edges and preserved more tissue details when compared with U-net-MARS. Conclusions:Both the CNN-MARS and U-net-MARS models developed in this study effectively remove the metal artifacts and enhance the image quality. CNN-MARS exhibited an advantage in restoring tissue structure information around the artifacts compared to U-net-MARS.

CRISPR/Cas9 has been widely used in engineering Saccharomyces cerevisiae for gene insertion, replacement and deletion due to its simplicity and high efficiency. The selectable markers of CRISPR/Cas9 systems are particularly useful for genome editing and Cas9-plasmids removing in yeast. In our previous research, GAL80 gene has been deleted by the plasmid pML104-mediated CRISPR/Cas9 system in an engineered yeast, in order to eliminate the requirement of galactose supplementation for induction. The maximum artemisinic acid production by engineered S. cerevisiae 1211-2 (740 mg/L) was comparable to that of the parental strain 1211 without galactose induction. Unfortunately, S. cerevisiae 1211-2 was inefficient in the utilization of the carbon source ethanol in the subsequent 50 L pilot fermentation experiment. The artemisinic acid yield in the engineered S. cerevisiae 1211-2 was only 20%-25% compared with that of S. cerevisiae 1211. The mutation of the selection marker URA3 was supposed to affect the growth and artemisinic acid production. A ura3 mutant was successfully restored by a recombinant plasmid pML104-KanMx4-u along with a 90 bp donor DNA, resulting in S. cerevisiae 1211-3. This mutant could grow normally in a fed-batch fermentor with mixed glucose and ethanol feeding, and the final artemisinic acid yield (> 20 g/L) was comparable to that of the parental strain S. cerevisiae 1211. In this study, an engineered yeast strain producing artemisinic acid without galactose induction was obtained. More importantly, it was the first report showing that the auxotrophic marker URA3 significantly affected artemisinic acid production in a pilot-scale fermentation with ethanol feeding, which provides a reference for the production of other natural products in yeast chassis.
Artemisinins ; Fermentation ; Saccharomyces cerevisiae/metabolism* ; Saccharomyces cerevisiae Proteins/metabolism*

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 determine the correlation of phosphorylated ribosomal S6 protein (P-S6) content in blood and brain tissue in mice and rats with seizure. METHODS: Seizure models were induced by intraperitoric injection of kainic acid (KA) in C57BL/mice and SD rats. Flow cytometry was used to detect the content of P-S6 in blood; Western blot was used to detect the expression of P-S6 in brain tissues. The correlation between P-S6 expression in blood and in brain tissue was examine by Pearson analysis, and the correlation between P-S6 expression in blood and the severity of seizure was also observed. RESULTS: Western blotting analysis showed that the expression of P-S6 was significantly increased in peripheral blood and brain tissue in mice 1 h after KA-induced seizure,and the expression levels increased to (1.49±0.45) times (<0.05) and (2.55±0.66) times ( <0.01) of the control group, respectively. Flow cytometry showed that the positive percentage and average fluorescence intensity of P-S6 in the blood of mice increased significantly 1 h after KA-induced seizures (<0.01), which was consistent with the expression of P-S6 in brain tissue (=0.8474, <0.01). Flow cytometry showed that the average fluorescence intensity of P-S6 in blood increased from 14.89±9.75 to 52.35±21.72 (<0.01) in rats with seizure, which was consistent with the change of P-S6 in brain tissue (=0.9385, <0.01). Rats with higher levels of seizure were of higher levels of P-S6 in peripheral blood. CONCLUSIONS Consistent correlation of P-S6 expression is demonstrated in peripheral blood and in brain tissue after KA-induced seizure, suggesting that the expression of P-S6 in blood can accurately reflect the changes of mTOR signaling pathway in brain tissue.
Animals ; Brain ; drug effects ; physiopathology ; Gene Expression Regulation ; drug effects ; Kainic Acid ; Mice ; Mice, Inbred C57BL ; Phosphorylation ; Rats ; Rats, Sprague-Dawley ; Seizures ; blood ; chemically induced ; physiopathology

Objective To investigate the values of combined detection of serum pancreas autoantibodies (PAB),anti-saccharomyces cerevisiae antibodies(ASCA),goblet cell autoantibodies(GAB) and antineutrophil cytoplasmic antibodies(PANCA) and fecal calprotectin(FC) in the diagnosis and differential diagnosis of inflammatory bowel diseases(IBD).Methods The serum and feces samples from IBD patients,including 107 with definite Crohn's disease(CD) and 98 with definite ulcerative colitis(UC),and 79 non-IBD patients as the control were collected.Serum PANCA,ASCA,GAB and PAB were detected by an indirect immunofluorescence assay,and FC concentration by double-antibody sandwich ELISA.The results from different patients were compared and analyzed.Results The positive rates of serum PANCA,GAB,PAB and ASCA in 205 IBD patients were 36.1％,29.8％,38.0％ and 4.9％,respectively.The FC concentrations in IBD,CD and UC patients were significantly higher than that in the control(P ＜ 0.01),while there was no statistical difference between CD and UC patients (P ＞ 0.05).The positive rates of PANCA in CD and UC patients were 8.4％ and 66.3％,respectively,while those of PAB in CD and UC patients were 65.4％ and 8.2％,respectively.The sensitivity and specificity of PAB,PANCA,GAB,ASCA,FC and their combination in the differential diagnosis of IBD and non-IBD were 38.0％,36.1％,29.8％,4.9％,54.1％,63.4％ and 98.7％,96.2％,94.9％,100％,68.4％,93.7％,respectively.The area under the ROC of the combination of 5 markers was 0.819 in differentially diagnosing IBD and non-IBD.The area under the ROC of PANCA for the differential diagnosis of UC was 0.816,while that of PAB for the differential diagnosis of CD was 0.823.Conclusion GAB is an autoantibody associated with IBD,which may be helpful for the auxiliary diagnosis of IBD.PAB and PANCA are the important serological markers for the diagnosis of CD and UC,respectively.The combination of FC with PAB,PANCA,GAB and ASCA may be used for the differential diagnosis of IBD and non-IBD,but has little value in distinguishing CD and UC.

A bicyclic depsipeptide, chromopeptide A (1), was isolated from a deep-sea-derived bacterium Chromobacterium sp. HS-13-94. Its structure was determined by extensive spectroscopic analysis and by comparison with a related known compound. The absolute configuration of chromopeptide A was established by X-ray diffraction analysis employing graphite monochromated Mo K α radiation (λ=0.71073 Å) with small Flack parameter 0.03. Chromopeptide A suppressed the proliferation of HL-60, K-562, and Ramos cells with average IC50 values of 7.7, 7.0, and 16.5 nmol/L, respectively.