1. MW-9, a chalcones derivative bearing heterocyclic moieties, ameliorates ulcerative colitis via regulating MAPK signaling pathway
Zhao WU ; Nan-Ting ZOU ; Chun-Fei ZHANG ; Hao-Hong ZHANG ; Qing-Yan MO ; Ze-Wei MAO ; Chun-Ping WAN ; Ming-Qian JU ; Chun-Ping WAN ; Xing-Cai XU
Chinese Pharmacological Bulletin 2024;40(3):514-520
		                        		
		                        			
		                        			 Aim To investigate the therapeutic effect of the MW-9 on ulcerative colitis(UC)and reveal the underlying mechanism, so as to provide a scientific guidance for the MW-9 treatment of UC. Methods The model of lipopolysaccharide(LPS)-stimulated RAW264.7 macrophage cells was established. The effect of MW-9 on RAW264.7 cells viability was detected by MTT assay. The levels of nitric oxide(NO)in RAW264.7 macrophages were measured by Griess assay. Cell supernatants and serum levels of inflammatory cytokines containing IL-6, TNF-α and IL-1β were determined by ELISA kits. Dextran sulfate sodium(DSS)-induced UC model in mice was established and body weight of mice in each group was measured. The histopathological damage degree of colonic tissue was assessed by HE staining. The protein expression of p-p38, p-ERK1/2 and p-JNK was detected by Western blot. Results MW-9 intervention significantly inhibited NO release in RAW264.7 macrophages with IC50 of 20.47 mg·L-1 and decreased the overproduction of inflammatory factors IL-6, IL-1β and TNF-α(P<0.05). MW-9 had no cytotoxicity at the concentrations below 6 mg·L-1. After MW-9 treatment, mouse body weight was gradually reduced, and the serum IL-6, IL-1β and TNF-α levels were significantly down-regulated. Compared with the model group, MW-9 significantly decreased the expression of p-p38 and p-ERK1/2 protein. Conclusions MW-9 has significant anti-inflammatory activities both in vitro and in vivo, and its underlying mechanism for the treatment of UC may be associated with the inhibition of MAPK signaling pathway. 
		                        		
		                        		
		                        		
		                        	
2.Exploring Scientific Connotation of "Fried Charcoal Survivability" of Lonicerae Japonicae Flos Based on Color-composition Correlation
Ting ZOU ; Jing WANG ; Xu WU ; Kai YANG ; Ming DANG ; Xiuchu GUO ; Lin WANG ; Chenxi LUO ; Juan PEI ; Chongbo ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(4):175-182
		                        		
		                        			
		                        			ObjectiveTo explore the scientific connotation of fried charcoal survivability of Lonicerae Japonicae Flos(LJF) by analyzing the correlation between the color change and the intrinsic components during the processing of LJF Carbonisata(LJFC), and taking pH, charcoal adsorption and microscopic characteristics as indexes. MethodLJFC samples with different degrees of processing were prepared according to the stir-frying time of 0.0, 1.5, 3.0, 4.5, 6.0, 7.5, 9.0, 10.5 min(numbered S1-S8), and the contents of gallic acid, chlorogenic acid, cryptochlorogenic acid, rutin, luteoloside, isochlorogenic acid A and isochlorogenic acid C were determined by high performance liquid chromatography(HPLC), and the L*(brightness), a*(red-greenness) and b*(yellow-blueness) of LJFC samples with different degrees of processing were determined by spectrophotometer, and the correlation analysis and principal component analysis(PCA) between the contents of seven representative components and the color of the samples were carried out by SPSS 26. 0 and SIMCA-P 14.1. Then pH, adsorption force and characteristic structure of different samples of LJFC were detected and the processing pattern of LJFC was analyzed. ResultThe results of quantitative analysis revealed that the contents of luteoloside, rutin, chlorogenic acid and isochlorogenic acid A gradually decreased, and the contents of cryptochlorogenic acid, isochlorogenic acid C and gallic acid firstly increased and then decreased. The L* and b* of the sample powders decreased, and a* showed a trend of increasing and then decreasing. The L* and b* were positively correlated with the contents of chlorogenic acid, rutin, luteoloside, isochlorogenic acid A, b* was positively correlated with the content of gallic acid, and a* was positively correlated with the contents of cryptochlorogenic acid and isochlorogenic acid C. PCA revealed that samples could be clearly divided into 3 groups, S1-S2 as one group, S3-S5 as one group, and S6-S8 as one group, with S3 having the highest score. The results of regression analysis showed that only isochlorogenic acid C could be used to predict the contents of components by colorimetric values combined with regression equations. Physicochemical analysis showed that pH of LJFC increased with the increase of degree of charcoal stir-frying, while adsorption force showed a tendency of increasing and then decreasing, with the highest adsorption force in the S5 sample, and the non-glandular hairs, calcium oxalate clusters and pollen grains had a varying degree of decreasing with the deepening of processing degree, and the microstructures of S6-S8 samples were obviously charred with pollen grains almost invisible. ConclusionThe changes in chemical composition and color characteristics of LJFC during the processing have certain correlations, combined with the changes in physicochemical properties, S5 sample is found to be the optimal processed products, which can provide a reference for the processing standardization and quality evaluation of LJFC, and enrich the scientific connotation of fried charcoal survivability of LJF. 
		                        		
		                        		
		                        		
		                        	
3.The Quantitative Evaluation of Automatic Segmentation in Lumbar Magnetic Resonance Images
Yao-Wen LIANG ; Yu-Ting FANG ; Ting-Chun LIN ; Cheng-Ru YANG ; Chih-Chang CHANG ; Hsuan-Kan CHANG ; Chin-Chu KO ; Tsung-Hsi TU ; Li-Yu FAY ; Jau-Ching WU ; Wen-Cheng HUANG ; Hsiang-Wei HU ; You-Yin CHEN ; Chao-Hung KUO
Neurospine 2024;21(2):665-675
		                        		
		                        			 Objective:
		                        			This study aims to overcome challenges in lumbar spine imaging, particularly lumbar spinal stenosis, by developing an automated segmentation model using advanced techniques. Traditional manual measurement and lesion detection methods are limited by subjectivity and inefficiency. The objective is to create an accurate and automated segmentation model that identifies anatomical structures in lumbar spine magnetic resonance imaging scans. 
		                        		
		                        			Methods:
		                        			Leveraging a dataset of 539 lumbar spinal stenosis patients, the study utilizes the residual U-Net for semantic segmentation in sagittal and axial lumbar spine magnetic resonance images. The model, trained to recognize specific tissue categories, employs a geometry algorithm for anatomical structure quantification. Validation metrics, like Intersection over Union (IOU) and Dice coefficients, validate the residual U-Net’s segmentation accuracy. A novel rotation matrix approach is introduced for detecting bulging discs, assessing dural sac compression, and measuring yellow ligament thickness. 
		                        		
		                        			Results:
		                        			The residual U-Net achieves high precision in segmenting lumbar spine structures, with mean IOU values ranging from 0.82 to 0.93 across various tissue categories and views. The automated quantification system provides measurements for intervertebral disc dimensions, dural sac diameter, yellow ligament thickness, and disc hydration. Consistency between training and testing datasets assures the robustness of automated measurements. 
		                        		
		                        			Conclusion
		                        			Automated lumbar spine segmentation with residual U-Net and deep learning exhibits high precision in identifying anatomical structures, facilitating efficient quantification in lumbar spinal stenosis cases. The introduction of a rotation matrix enhances lesion detection, promising improved diagnostic accuracy, and supporting treatment decisions for lumbar spinal stenosis patients. 
		                        		
		                        		
		                        		
		                        	
4.The Quantitative Evaluation of Automatic Segmentation in Lumbar Magnetic Resonance Images
Yao-Wen LIANG ; Yu-Ting FANG ; Ting-Chun LIN ; Cheng-Ru YANG ; Chih-Chang CHANG ; Hsuan-Kan CHANG ; Chin-Chu KO ; Tsung-Hsi TU ; Li-Yu FAY ; Jau-Ching WU ; Wen-Cheng HUANG ; Hsiang-Wei HU ; You-Yin CHEN ; Chao-Hung KUO
Neurospine 2024;21(2):665-675
		                        		
		                        			 Objective:
		                        			This study aims to overcome challenges in lumbar spine imaging, particularly lumbar spinal stenosis, by developing an automated segmentation model using advanced techniques. Traditional manual measurement and lesion detection methods are limited by subjectivity and inefficiency. The objective is to create an accurate and automated segmentation model that identifies anatomical structures in lumbar spine magnetic resonance imaging scans. 
		                        		
		                        			Methods:
		                        			Leveraging a dataset of 539 lumbar spinal stenosis patients, the study utilizes the residual U-Net for semantic segmentation in sagittal and axial lumbar spine magnetic resonance images. The model, trained to recognize specific tissue categories, employs a geometry algorithm for anatomical structure quantification. Validation metrics, like Intersection over Union (IOU) and Dice coefficients, validate the residual U-Net’s segmentation accuracy. A novel rotation matrix approach is introduced for detecting bulging discs, assessing dural sac compression, and measuring yellow ligament thickness. 
		                        		
		                        			Results:
		                        			The residual U-Net achieves high precision in segmenting lumbar spine structures, with mean IOU values ranging from 0.82 to 0.93 across various tissue categories and views. The automated quantification system provides measurements for intervertebral disc dimensions, dural sac diameter, yellow ligament thickness, and disc hydration. Consistency between training and testing datasets assures the robustness of automated measurements. 
		                        		
		                        			Conclusion
		                        			Automated lumbar spine segmentation with residual U-Net and deep learning exhibits high precision in identifying anatomical structures, facilitating efficient quantification in lumbar spinal stenosis cases. The introduction of a rotation matrix enhances lesion detection, promising improved diagnostic accuracy, and supporting treatment decisions for lumbar spinal stenosis patients. 
		                        		
		                        		
		                        		
		                        	
5.Downregulation of MUC1 Inhibits Proliferation and Promotes Apoptosis by Inactivating NF-κB Signaling Pathway in Human Nasopharyngeal Carcinoma
Shou-Wu WU ; Shao-Kun LIN ; Zhong-Zhu NIAN ; Xin-Wen WANG ; Wei-Nian LIN ; Li-Ming ZHUANG ; Zhi-Sheng WU ; Zhi-Wei HUANG ; A-Min WANG ; Ni-Li GAO ; Jia-Wen CHEN ; Wen-Ting YUAN ; Kai-Xian LU ; Jun LIAO
Progress in Biochemistry and Biophysics 2024;51(9):2182-2193
		                        		
		                        			
		                        			ObjectiveTo investigate the effect of mucin 1 (MUC1) on the proliferation and apoptosis of nasopharyngeal carcinoma (NPC) and its regulatory mechanism. MethodsThe 60 NPC and paired para-cancer normal tissues were collected from October 2020 to July 2021 in Quanzhou First Hospital. The expression of MUC1 was measured by real-time quantitative PCR (qPCR) in the patients with PNC. The 5-8F and HNE1 cells were transfected with siRNA control (si-control) or siRNA targeting MUC1 (si-MUC1). Cell proliferation was analyzed by cell counting kit-8 and colony formation assay, and apoptosis was analyzed by flow cytometry analysis in the 5-8F and HNE1 cells. The qPCR and ELISA were executed to analyze the levels of TNF-α and IL-6. Western blot was performed to measure the expression of MUC1, NF-кB and apoptosis-related proteins (Bax and Bcl-2). ResultsThe expression of MUC1 was up-regulated in the NPC tissues, and NPC patients with the high MUC1 expression were inclined to EBV infection, growth and metastasis of NPC. Loss of MUC1 restrained malignant features, including the proliferation and apoptosis, downregulated the expression of p-IкB、p-P65 and Bcl-2 and upregulated the expression of Bax in the NPC cells. ConclusionDownregulation of MUC1 restrained biological characteristics of malignancy, including cell proliferation and apoptosis, by inactivating NF-κB signaling pathway in NPC. 
		                        		
		                        		
		                        		
		                        	
6.Development and Application of Detection Methods for Capture and Transcription Elongation Rate of Bacterial Nascent RNA
Yuan-Yuan LI ; Yu-Ting WANG ; Zi-Chun WU ; Hao-Xuan LI ; Ming-Yue FEI ; Dong-Chang SUN ; O. Claudio GUALERZI ; Attilio FABBRETTI ; Anna Maria GIULIODORI ; Hong-Xia MA ; Cheng-Guang HE
Progress in Biochemistry and Biophysics 2024;51(9):2249-2260
		                        		
		                        			
		                        			ObjectiveDetection and quantification of RNA synthesis in cells is a widely used technique for monitoring cell viability, health, and metabolic rate.After exposure to environmental stimuli, both the internal reference gene and target gene would be degraded. As a result, it is imperative to consider the accurate capture of nascent RNA and the detection of transcriptional levels of RNA following environmental stimulation. This study aims to create a Click Chemistry method that utilizes its property to capture nascent RNA from total RNA that was stimulated by the environment. MethodsThe new RNA was labeled with 5-ethyluridine (5-EU) instead of uracil, and the azido-biotin medium ligand was connected to the magnetic sphere using a combination of “Click Chemistry” and magnetic bead screening. Then the new RNA was captured and the transcription rate of 16S rRNA was detected by fluorescence molecular beacon (M.B.) and quantitative reverse transcription PCR (qRT-PCR). ResultsThe bacterial nascent RNA captured by “Click Chemistry” screening can be used as a reverse transcription template to form cDNA. Combined with the fluorescent molecular beacon M.B.1, the synthesis rate of rRNA at 37℃ is 1.2 times higher than that at 15℃. The 16S rRNA gene and cspI gene can be detected by fluorescent quantitative PCR,it was found that the measured relative gene expression changes were significantly enhanced at 25℃ and 16℃ when analyzed with nascent RNA rather than total RNA, enabling accurate detection of RNA transcription rates. ConclusionCompared to other article reported experimental methods that utilize screening magnetic columns, the technical scheme employed in this study is more suitable for bacteria, and the operation steps are simple and easy to implement, making it an effective RNA capture method for researchers. 
		                        		
		                        		
		                        		
		                        	
7.FHL2 interacts with LDHA to promote glioma cell proliferation
WU Guoqing ; ZHANG Ting△ ; SONG Xiaofeng ; ZHU Ting ; LI Na ; LI Ming
Chinese Journal of Cancer Biotherapy 2024;31(10):976-983
		                        		
		                        			
		                        			[摘  要]  目的:探讨四个半LIM结构域2(FHL2)蛋白对胶质瘤细胞增殖的影响及其分子机制。方法:利用TCGA和CGGA数据库分析胶质瘤组织中FHL2 mRNA表达水平与患者预后的关系。通过WB法检测人胶质瘤组织标本及人胶质瘤细胞U87、T98G、U251、SNB19、GSC23、A172、LN229、G267和星形胶质细胞NHA中的FHL2蛋白表达水平。利用慢病毒载体构建稳定敲低FHL2的U87细胞和过表达FHL2的SNB19细胞,即U87-shGFP、U87-shFHL2-1#、U87-shFHL2-4#和SNB19-3flag、SNB19-3flag-FHL2组。通过CCK-8法、克隆形成实验检测敲低和过表达FHL2对细胞增殖的影响,免疫共沉淀(Co-IP)和液相色谱-串联质谱(LC/MS)法筛选FHL2在胶质瘤细胞中的相互作用蛋白,并用Co-IP和免疫荧光法验证它们的结合作用和共定位情况。使用酶标仪检测敲低和过表达FHL2细胞内乳酸产量和乳酸脱氢酶(LDH)活性的变化,WB法分析FHL2、LDHA及p-LDHA在正常脑组织和胶质瘤组织中的蛋白表达差异及其相互关系。在过表达 FHL2的SNB19细胞中使用LDHA的小分子抑制剂AT-101,通过CCK-8实验和酶标仪比色法验证FHL2在胶质瘤乳酸代谢中的作用,验证AT-101在胶质瘤中潜在的治疗效果。结果:Co-IP和LC/MS检测发现,FHL2与LDHA在胶质瘤细胞中存在相互作用。FHL2过表达可提高LDHA活性和乳酸生成(均P < 0.001),进而促进胶质瘤细胞增殖(P < 0.001)。相反,敲低FHL2会降低LDHA活性和乳酸产量(P < 0.001或P < 0.05)并抑制细胞增殖(P < 0.001)。AT101能抑制LDHA活性,并显著抑制FHL2促进胶质瘤细胞的增殖,同时恢复磷酸化LDHA(Y10)水平(P<0.01或P < 0.001)。结论:FHL2与LDHA蛋白相互作用,FHL2通过激活p-LDHA(Y10)的表达促进LDHA活性和乳酸产生,进而促进胶质瘤细胞的增殖,靶向这种相互作用可能成为治疗胶质瘤的潜在策略。
		                        		
		                        		
		                        		
		                        	
8.Teaching Reform of “Structural Biology” Course Based on The Ability Cultivation
Ying-Jie LI ; Ting-Ting GUO ; Ming-Yu WANG ; Da-Lei WU ; Xiang GAO ; Lu-Shan WANG
Progress in Biochemistry and Biophysics 2024;51(11):3046-3053
		                        		
		                        			
		                        			As a rapidly developing frontier discipline, structural biology has penetrated into every field of life science research. The course of “Structural Biology” plays an important role in expanding the knowledge system of undergraduate students and promoting students’ scientific spirit and innovation. For the high-quality training of highly skilled talents, we aimed to promote the original innovation of students, the ability of thinking, and the ability of engineering practice. The trinity education concept, including shape of the value, passing on knowledge, and ability cultivation, was applied. During the reform, we explored a step-by-step course content and searched for factors involved in ideological and political education. Based on the problem-based learning (PBL) method, a hybrid teaching model was designed to cultivate the problem-thinking and problem-solving skills of students. Meanwhile, a number of evaluation systems for students and teachers were established, which may be generally adopted for the course of “Structural Biology”. The survey data suggested that the exploration has a good effect on teaching and training and is conducive to the cultivation of research-oriented, comprehensive, innovative talents under the background of “New Engineering”. 
		                        		
		                        		
		                        		
		                        	
9.Relationship between Phenotypic Changes of Dendritic Cell Subsets and the Onset of Plateau Phase during Intermittent Interferon Therapy in Patients with CHB
Liu YANG ; Yu Shi WANG ; Ting Ting JIANG ; Wen DENG ; Min CHANG ; Ling Shu WU ; Hua Wei CAO ; Yao LU ; Ge SHEN ; Yu Ru LIU ; Jiao Yuan GAO ; Jiao Meng XU ; Ping Lei HU ; Lu ZHANG ; Yao XIE ; Hui Ming LI
Biomedical and Environmental Sciences 2024;37(3):303-314
		                        		
		                        			
		                        			Objective This study aimed to evaluate whether the onset of the plateau phase of slow hepatitis B surface antigen decline in patients with chronic hepatitis B treated with intermittent interferon therapy is related to the frequency of dendritic cell subsets and expression of the costimulatory molecules CD40,CD80,CD83,and CD86. Method This was a cross-sectional study in which patients were divided into a natural history group(namely NH group),a long-term oral nucleoside analogs treatment group(namely NA group),and a plateau-arriving group(namely P group).The percentage of plasmacytoid dendritic cell and myeloid dendritic cell subsets in peripheral blood lymphocytes and monocytes and the mean fluorescence intensity of their surface costimulatory molecules were detected using a flow cytometer. Results In total,143 patients were enrolled(NH group,n = 49;NA group,n = 47;P group,n = 47).The results demonstrated that CD141/CD1c double negative myeloid dendritic cell(DNmDC)/lymphocytes and monocytes(%)in P group(0.041[0.024,0.069])was significantly lower than that in NH group(0.270[0.135,0.407])and NA group(0.273[0.150,0.443]),and CD86 mean fluorescence intensity of DNmDCs in P group(1832.0[1484.0,2793.0])was significantly lower than that in NH group(4316.0[2958.0,5169.0])and NA group(3299.0[2534.0,4371.0]),Adjusted P all<0.001. Conclusion Reduced DNmDCs and impaired maturation may be associated with the onset of the plateau phase during intermittent interferon therapy in patients with chronic hepatitis B.
		                        		
		                        		
		                        		
		                        	
10.Association of Cytokines with Clinical Indicators in Patients with Drug-Induced Liver Injury
Hua Wei CAO ; Ting Ting JIANG ; Ge SHEN ; Wen DENG ; Yu Shi WANG ; Yu Zi ZHANG ; Xin Xin LI ; Yao LU ; Lu ZHANG ; Yu Ru LIU ; Min CHANG ; Ling Shu WU ; Jiao Yuan GAO ; Xiao Hong HAO ; Xue Xiao CHEN ; Ping Lei HU ; Jiao Meng XU ; Wei YI ; Yao XIE ; Hui Ming LI
Biomedical and Environmental Sciences 2024;37(5):494-502
		                        		
		                        			
		                        			Objective To explore characteristics of clinical parameters and cytokines in patients with drug-induced liver injury(DILI)caused by different drugs and their correlation with clinical indicators. Method The study was conducted on patients who were up to Review of Uncertainties in Confidence Assessment for Medical Tests(RUCAM)scoring criteria and clinically diagnosed with DILI.Based on Chinese herbal medicine,cardiovascular drugs,non-steroidal anti-inflammatory drugs(NSAIDs),anti-infective drugs,and other drugs,patients were divided into five groups.Cytokines were measured by Luminex technology.Baseline characteristics of clinical biochemical indicators and cytokines in DILI patients and their correlation were analyzed. Results 73 patients were enrolled.Age among five groups was statistically different(P=0.032).Alanine aminotransferase(ALT)(P=0.033)and aspartate aminotransferase(AST)(P=0.007)in NSAIDs group were higher than those in chinese herbal medicine group.Interleukin-6(IL-6)and tumor necrosis factor alpha(TNF-α)in patients with Chinese herbal medicine(IL-6:P<0.001;TNF-α:P<0.001)and cardiovascular medicine(IL-6:P=0.020;TNF-α:P=0.001)were lower than those in NSAIDs group.There was a positive correlation between ALT(r=0.697,P=0.025),AST(r=0.721,P=0.019),and IL-6 in NSAIDs group. Conclusion Older age may be more prone to DILI.Patients with NSAIDs have more severe liver damage in early stages of DILI,TNF-α and IL-6 may partake the inflammatory process of DILI.
		                        		
		                        		
		                        		
		                        	
            
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