1.Effect of Modified Gegen Qinlian Decoction (加味葛根芩连汤) on the Intestinal Mucus Barrier and Intestinal Stem Cell Proliferation and Differentiation in Ulcerative Colitis Model Mice
Jinke HUANG ; Jiaqi ZHANG ; Fengyun WANG ; Xudong TANG
Journal of Traditional Chinese Medicine 2025;66(9):941-947
ObjectiveTo explore the possible mechanism of Modified Gegen Qinlian Decoction (加味葛根芩连汤, MGQD) in the treatment of ulcerative colitis (UC) based on intestinal mucus barrier. MethodsThirty C57BL/6 mice were randomly divided into a control group, a model group and a MGQD group with 10 mice in each. Dextran Sulfate Sodium Salt (DSS) was used to construct the UC model in all groups except for the control group. Meanwhile, mice in the MGQD group were given 20 g/kg of MGQD decoction by gavage according to their body weight, while those in the control group and model group were given 0.2 ml/20 g of pure water by gavage, once a day for 7 consecutive days. On the day following the last gavage, the body weight, disease activity index (DAI) score, spleen weight, and colon length were compared. The pathological changes of the intestinal mucosal tissues were observed by HE staining; the protein expression levels of mucin 2 (MUC2) and leucine-rich repeat G protein-coupled receptor 5 (Lgr5) in the intestinal mucosal tissues were detected by immunofluorescence; the cuprocytes in the intestinal mucosal tissues were detected by AB/PAS staining; and the expression level of Ki67 in the intestinal mucosal tissues was detected by immunohistochemistry. ResultsHE staining showed that the colon mucosal tissue of the mice in the control group was intact. In the model group, the colon mucosal epithelial structure was severely damaged, with a large amount of inflammatory cell infiltration in the mucosal propria. In the MGQD group, the mucosal tissue structure was partially lost, with a small amount of inflammatory cell infiltration.The body weight and colon length of mice in the model group decreased significantly compared to those in the control group, while DAI scores and spleen weight increased, and the levels of MUC2, Ki67, Lgr5 proteins, and the number of goblet cells were significantly reduced (P<0.01). Compared to the model group, the MGQD group had increased body weight of mice, colon length, and decreased DAI scores and spleen weight; the levels of MUC2, Ki67, Lgr5 proteins, and the number of goblet cells were increased (P<0.05 or P<0.01). ConclusionMGQD has a favorable ameliorative effect on UC-related symptoms and pathological tissue damage, and its mechanism of action may be related to the restoration of the prolife-ration and differentiation of intestinal stem cells into goblet cells, thereby promoting the repair of the intestinal mucus barrier.
2.Cell-of-origin for heterotopic ossification induced by bone morphogenetic protein 4 in skeletal muscle
Yangyi YU ; Qiang LIAN ; Jianqun WU ; Xuan ZHANG ; Jinke REN ; Guangheng LI
Chinese Journal of Tissue Engineering Research 2024;28(25):4034-4040
BACKGROUND:Heterotopic ossification of skeletal muscle is a clinically serious complication.For heterotopic ossification of skeletal muscles,the cells involved in the process of heterotopic ossification remain unclear. OBJECTIVE:To investigate the involvement of myocytes,fascia cells,and endothelial cells in the process of heterotopic ossification in skeletal muscle and to observe the cell origin of heterotopic ossification in skeletal muscle induced by bone morphogenetic protein 4. METHODS:Both C2C12 cells and the myotubes formed by the C2C12 cells in the induction medium were cultured,and 500 ng/mL bone morphogenetic protein 4 was added to the medium respectively,and whether the C2C12 cells and myotubes continued to proliferate within 10 days under the treatment were observed under a microscope.Myogenic cells(L6,derived from rats)and fibroblast-derived cells(derived from human)were co-cultured.After treatment with 500 ng/mL bone morphogenetic protein 4 and 10 ng/mL transforming growth factor-β,osteogenic and chondrogenic differentiation potential within 21 days were observed using Safranine O staining and Alcian blue staining.Using transgenic animal FVB/N-TgN(TIE2-LacZ)182Sato mice,15 μL of adeno-associated virus-bone morphogenetic protein 4(5×1010 PFU/mL)were implanted in the thigh muscle space of genetic mice for 10 and 14 days.X-gal staining was used to observe the formation of new blood vessel endothelium in the differentiated bone. RESULTS AND CONCLUSION:(1)Bone morphogenetic protein 4 caused myotube breakdown and increased C2C12 cell proliferation.Compared with other groups,the pure fibroblast-derived cell group had a higher area of positive alcian blue and safarin O staining(P<0.05)and a lower area of alkaline phosphatase staining(P<0.05),while the pure L6 group had a bigger area of alkaline phosphatase staining(P<0.05)but a smaller area of positive alcian blue and safarin O staining(P<0.05).(2)Transplantation of adeno-associated virus-bone morphogenetic protein 4-adsorbed gelatin sponge into FVB/N-TgN(TIE2-LacZ)182Sato mice resulted in heterotopic ossification.(3)X-gal staining results demonstrated that there was no obvious staining in chondrocytes and differentiated bones and Tie2+ endothelial cells did not participate in the formation of the alienated bone.(4)These findings verify that fibroblasts are the primary source of osteoblasts during the adeno-associated virus-bone morphogenetic protein 4-induced ectopic endochondral ossification in skeletal muscle,but myogenic cells are the main source of osteoblasts.Tie2+ endothelial cells might not be the cell source for cartilage and bone.
3.Effects of Modified Gegen Qinlian Decoction (葛根芩连汤) on Intestinal Mucosal Barrier and Epithelial Mesenchymal Transition in Ulcerative Colitis Model Mice
Lihui FANG ; Jiaqi ZHANG ; Lanshuo HU ; Xintong WANG ; Shan LIU ; Yuedan WANG ; Jinke HUANG ; Xudong TANG
Journal of Traditional Chinese Medicine 2024;65(24):2580-2588
ObjectiveTo investigate the possible mechanism of modified Gegen Qinlian Decoction (葛根芩连汤) in treatment of ulcerative colitis (UC) from the view of intestinal mucosal epithelial barrier damage and epithelial mesenchymal transition. MethodsSixty male C57BL/6J mice were divided into blank group, model group, western medicine control group, and low-, medium-, and high-dose modified Gegen Qinlian Decoction groups, with 10 mice in each group. Except for the blank group, 3% dextran sodium sulfate (DSS) was used to induce colitis model by free drinking for 7 days, and on the first day of modelling, 6, 12, and 24 g/(kg·d) of modified Gegen Qinlian Decoction were given to the low-, medium-, and high-dose groups respectively, 5-aminosalicylic acid (5-ASA) 100 mg/(kg·d) given by gavage to western medicine control group, and 10 ml/kg distilled water were given to blank and model group by gavage, once a day for 7 days. Body mass of mice was recorded and disease activity index (DAI) scores were performed daily. The mice were anesthetized after 24h of the last administration and the colon was taken to observe the length of colon, HE staining was applied to observe the damage of colonic mucosa and score pathological states, Masson staining to detect the deposition of colonic collagen fibers, immunofluorescence to observe the distribution of F-actin in colonic mucosal epithelium, and immunohistochemistry to detect the expression of tight junction protein ZO-1, Occludin, E-cadherin and Vimentin. ResultsCompared with the blank group at the same time, the percentage of body mass of mice in the model group on day 7 of modelling significantly reduced and the DAI score was significantly increased (P<0.01); compared with the model group at the same time, the body mass of mice in the western medicine control group and all of modified Gegen Qinlian Decoction groups decreased, and the DAI scores of mice in the western medicine control group and the high-dose modified Gegen Qinlian Decoction group decreased (P<0.05 or P<0.01); compared with the same time of mice in the low-dos Gegen Qinlian Decoction group, the body mass of mice in the high-dose Gegen Qinlian Decoction group and the western medicine control group significantly elevated (P<0.05). Compared with the blank group, the length of the colon of mice in the model group was significantly shortened, the pathological score and the percentage of collagen area were significantly increased, the average fluorescence intensity of F-actin was reduced, the protein levels of ZO-1, Occludin and E-cadherin in the colon tissue decreased, and the protein level of Vimentin elevated (P<0.01). Compared with the model group, the length of colon significantly increased, patholo-gical score, collagen area percentage decreased, ZO-1, Occludin, E-cadherin protein levels increased and Vimentin levels decreased in all medicated groups; the average fluorescence intensity of F-actin increased in the western medicine control group and the middle- and high-dose Gegen Qinlian Decoction groups (P<0.05 or P<0.01). Compared with the low-dose Gegen Qinlian Decoction group, the proportion of collagen fibre area in the middle-dose Gegen Qinlian Decoction group and the western medicine control group reduced; the mean fluorescence intensity of F-actin increased in the middle-dose Gegen Qinlian Decoction group; the protein levels of ZO-1 and E-cadherin increased in the western medicine control group, and the protein levels of ZO-1 increased in the high-dose Gegen Qinlian Decoction group (P<0.05). Compared with the medium-dose Gegen Qinlian Decoction group, the protein levels of ZO-1 elevated in the high-dose Gegen Qinlian Decoction group (P<0.05). Comapred with the high-dose Gegen Qinlian Decoction group, level of E-cadherin and Vimentin protein of the western medicine control group increased (P<0.05). ConclusionModified Gegen Qinlian Decoction was able to reduce colonic inflammation and mucosal barrier damage and inhibit the process of epithelial mesenchymal transition in mice models of ulcerative colitis, which may be one of its action mechanisms .
4.Integrating Network Pharmacology Based on UPLC-Q-Exactive/MS Technology to Explore the Mechanism of Chaihu Guizhi Decoction in the Treatment of Secondary Bacterial Pneumonia Caused by Influenza
Yuxiu HAN ; Jing ZHANG ; Junyu LUO ; Yanting JIA ; Jinke XU ; Qihui SUN ; Xu WANG ; Yong YANG ; Rong RONG
World Science and Technology-Modernization of Traditional Chinese Medicine 2023;25(6):2111-2121
Objective To study the mechanism of Chaihu Guizhi Decoction(CGD)in the treatment of influenza and staphylococcus aureus co-infection.Methods The co-infection model of influenza and staphylococcus aureus was established and CGD was used to intervene.The chemical components of CGD were qualitatively analyzed by UPLC-Q-Exactive/MS technology.The potential action targets of chemical components in CGD and the related targets of influenza Staphylococcus aureus co-infection were mined by network pharmacology method.The"component target disease"network was constructed.Core targets were selected according to degree ranking.Core action pathways were enriched by KEGG analysis and GO annotation analysis.The core target was verified by RT-qPCR,and the interaction between the core component and the key target was verified by molecular docking.Results CGD could significantly improve the decrease of body weight and thymus index(P<0.05)caused by co-infection.The lung index(P<0.05),relative amount of MmRNA expression(P<0.05)and bacterial load(P<0.05)were decreased,and the survival rate was improved.51 chemical constituents were identified from CGD.Through network pharmacological analysis,107 related targets corresponding to CGD treatment of bacterial pneumonia secondary to influenza were excavated.TNF,AKT1,ALB,VEGFA,MAPK3,PTGS2,STAT3,EGFR and other targets with strong correlation,mainly involved Fc epsilon RI signal pathway,GnRH signal pathway,NF-κB signal path,etc.Molecular docking study showed that the main active component of CGD,including oroxyloside,baicalein and wogonin have strong affinity with TNF,PTGS2 and EGFR targets.Compared with co-infection model group,in CGD group TNF-α、EGFR and PTGS2 increased significantly(P<0.05).Conclusion The main active ingredient of CGD is oroxyloside,baicalein and wogonin.TNF-α,PTGS2,EGFR and other targets to played a role in the treatment of influenza staphylococcus aureus co-infection.
5.A calibration curve model based on sine function
Xianmin ZHUANG ; Hanlin PAN ; Xi LIU ; Ning MA ; Jinke ZHANG ; Shuai ZHANG ; Guangmin QIAO ; Xiaoxia DU ; Yan HUANG ; Yanchao LI ; Zheng WANG
Chinese Journal of Laboratory Medicine 2023;46(9):934-942
Objective:This paper is to propose a calibration model based on sine function which enables more choices to determine the functional relationship between the absorbance and the concentration of the tested substance.Methods:This paper uses Taylor series expansion and Levenberg-Marquardt to obtain the optimal parameters for the Sine model and then summarizes the characters of the Sine model. On the basis of these characters, this paper compares and evaluates the experimental data processed by the Sine model from four aspects: correctness, precision, linearity and correlation.Results:The generated sine function calibration model achieved deviations within ±3% of the national standard substance, precision ( CV%) less than 2%, and a linear correlation coefficient greater than 0.990 within the measurement range of 32-710 mg/L. The correlation coefficients between the sine model and other well-performing linear calibration models for 104 clinical samples were all greater than 0.990. Conclusions:The performance evaluation of the prealbumin assay kit using the sine function calibration model meets industry standards and shows good correlation with the results of clinical sample measurements. This indicates that the sine function calibration model can serve as a new calibration model for in vitro diagnostic research and clinical applications.
6.Rapid Identification and Determination of Polysaccharides Contents in Anoectochilus Roxburghii Based on Near Infrared Spectroscopy with Chemometrics
ZHANG Xun ; HUANG Xiaoxuan ; YIN Jinke ; CHEN Yancheng ; LIN Yu ; WANG Xiaoying ; XU Wen
Chinese Journal of Modern Applied Pharmacy 2023;40(19):2702-2712
OBJECTIVE To distinguish Anoectochilus roxburghii and relative species by near infrared(NIR) spectroscopy combined with chemometrics, and to establish a prediction model for rapid determine polysaccharides contents in Anoectochilus roxburghii. METHODS The NIR spectroscopy of Anoectochilus roxburghii, Anoectochilus formosanus Hayata and Ludisia discolor were collected. The prepossessing of original spectrum was optimization through accuracy of classification in the NIR model, and six supervised pattern recognition algorithms such as decision tree, K-nearest neighbor algorithm, random forest, partial least squares regression discriminant analysis, linear discriminant analysis and support vector machine(SVM) were applied to identify effect of the classification effect, optimum algorithm and then establish qualitative model. The content of polysaccharides in 76 batches of Anoectochilus roxburghii samples were examined by ultraviolet visible spectrophotometry combined with phenol sulfuric acid method. In order to select optimization algorithm, six quantitative stoichiometry algorithms consisted of SVM, extreme learning machines, decision trees, random forests, principal component regression and partial least squares regression(PLS) were used to connect polysaccharide content and the NIR spectroscopy in Anoectochilus roxburghii respectively. The best method for determining the content of Anoectochilus roxburghii polysaccharides was further optimized by spectra pretreatment, band selection and number of band variables based on successive projection algorithm(SPA). RESULTS The NIR discriminant analysis model was established by SVM with SNV+SG+2ndD, and the classification accuracy was best. The prediction performance was evaluated based on the radial basis kernel function algorithm combined with confusion matrix and ROC curve, and the model performance was good. In addition, the quantitative analysis model was constructed by continuous projection-partial least squares by the prepossessing of SNV+SG+2ndD and the optimal band of 7 000-4 000 cm-1 with 97 of variables number. The accuracy was 0.992, which was the highest. The root mean square error calibration set, correlation coefficient of calibration set, and the root mean square error in validation set, correlation coefficient of validation set were 0.625, 0.993, 0.767, 0.992, separately. The prediction deviation was 8.467, and relative deviation of prediction set was less than 10%. CONCLUSION The established NIR-SVM qualitative model and SPA-PLS quantitative model are accurate and reliable, which are enable to identify Anoectochilus roxburghii and determine polysaccharide content nondestructively. It is a new and promising method for rapid evaluation of Anoectochilus roxburghii quality.
7.Factors affecting the motivation for weight self-management among pregnant women
Jinke CHANG ; Junhui ZHANG ; Wenfang ZHAO ; Aiju LIU
Journal of Preventive Medicine 2022;34(9):946-950
Objective:
To investigate the motivation for weight self-management and analyze its influencing factors among pregnant women in a tertiary hospital in Hohhot City, Inner Mongolia Autonomous Region, so as to provide insights into body weight management during pregnancy.
Methods:
Pregnant women at ages of 18 years and older that underwent prenatal examinations in a tertiary hospital in Hohhot City from January to March, 2022 were sampled using a convenience sampling method. Subjects' age, height, weight, occupation, residence, monthly household income, history of gestation and childbirth and gestational period were collected, and the motivation for weight self-management was evaluated among pregnant women using a pregnancy weight management protein motivation scale. The factors affecting the motivation for weight self-management were identified among pregnant women using a multivariable linear regression model.
Results:
Totally 969 pregnant women were recruited, including 841 women at ages of <35 years (86.79%), 780 women with an educational level of diploma and above (80.50%), 794 women living in urban areas (81.94%), 729 primiparas (75.23%), 421 women in the third trimester of gestation (43.45%) and 758 women with pre-pregnancy body mass index (BMI) of <25 kg/m2 (78.22%). The mean score of motivation for weight self-management was 127.53±14.60 among the pregnant women. Multivariable linear regression analysis showed that an educational level of high school and below (β′=-0.201), unemployed/self-employed individuals (β′=-0.077), living in rural areas (β′=-0.059), monthly household income of <10 000 yuan (<5 000 yuan, β′=-0.238; 5 000 to 10 000 yuan, β′=-0.169), in the third trimester of gestation (β′=-0.135), pre-pregnancy BMI of 25 kg/m2 and higher (β′=-0.214) and reduced the motivation for weight self-management among pregnant women.
Conclusion
The motivation for weight self-management correlates with gestational period, pre-pregnancy BMI, residence, occupation, educational level and monthly household income among pregnant women.
8.Research progress on nutrition management of kidney transplantation recipients
Yan TANG ; Jinke LI ; Tao QIU ; Long ZHANG ; Mengdan SHENG ; Ling CHEN
Chinese Journal of Modern Nursing 2022;28(20):2795-2800
Kidney transplantation is the most fundamental treatment for patients with end-stage renal disease. However, due to the long-term malnutrition of the patient before transplantation, coupled with the use of stress stimulation, immunosuppressive agents, and high-dose hormones after kidney transplantation, the patient is in a hypermetabolic state, and malnutrition is aggravated. The nutrition status of patients has a significant impact on the prognosis and development of kidney transplantation. Therefore, this article reviews the current status of nutrition intervention, the assessment of malnutrition, the related factors of malnutrition, and nutrition management in kidney transplantation recipients, so as to provide a reference for the nutrition and diet management of kidney transplantation recipients.
9.Treatment of large segmental humeral defects with unilateral external fixation and bone transport
Tinghui XIAO ; Yimiao LIN ; Jinke REN ; Zhaofeng JIA ; Hua WANG ; Guangheng LI ; Xiaoming ZHANG
Chinese Journal of Orthopaedic Trauma 2020;22(11):997-1000
Objective:To investigate the clinical treatment of large segmental humeral defects with unilateral external fixation and bone transport.Methods:A retrospective study was conducted of the 9 patients who had been treated at Department of Orthopedics, Shenzhen People's Hospital for large segmental humeral defects from September 2017 to June 2019. They were 5 males and 4 females with an average age of 29 years (from 21 to 41 years). Their defects were caused by trauma in 2 cases, by chronic osteomyelitis in 6 cases and by bone tumor in one case. The length of bone defect ranged from 4.2 to 9.0 cm, with an average of 5.9 cm. A unilateral external fixator was placed in operation, and adjusted regularly 7 to 10 days after operation for bone transport and bone lengthening to restore the length of humerus gradually. The external fixation bracket was removed after 3 to 4 layers of cortex were observed on X-ray films. Recorded were length and rate of humeral lengthening, fracture healing time, time for carrying external fixator and complications; the Disabilities of the Arm, Shoulder and Hand (DASH) scores were compared between preoperation and 15 months postoperation.Results:All the patients were followed up for 15 to 36 months (mean, 19 months). The length of lengthening averaged 5.9 cm (from 4.2 to 9.0 cm) with an average lengthening rate of 26%, the healing index 31 d/cm, the bone healing time 8.3 months, and the time for carrying external fixator 10.8 months(from 8.0 to 13.5 months). Their average DASH scores improved significantly from 25.0 ± 2.4 preoperation to 12.0 ± 1.8 at 15 months postoperation ( P<0.05). Good correction of large humeral defects was achieved in all but one case who reported temporary radial nerve paralysis. There were no such complications as neurovascular injury. The shoulder and elbow functions were basically normal after operation. Conclusions:In the treatment of large segmental humeral defects, unilateral external fixation plus bone transport can quickly repair the defects and recover the upper limb function of the patients.
10.The value of texture analysis based on T 2WI and apparent diffusion coefficient map in discriminating low grade from high grade prostate cancer
Jinke XIE ; Xiangde MIN ; Basen LI ; Zhaoyan FENG ; Peipei ZHANG ; Wei CAI ; Huijuan YOU ; Chanyuan FAN ; Liang WANG
Chinese Journal of Radiology 2020;54(12):1191-1196
Objective:To investigate the value of texture analysis based on T 2WI and apparent diffusion coefficient (ADC) maps in discriminating low grade from high grade prostate cancer (PCa). Methods:Retrospective analysis was performed on patients who were confirmed to be PCa by pathology after surgery and underwent MRI examination in the department of radiology,Tongji Hospital,Tongji Medical College, Huazhong University of Science and Technology before radical surgery, including routine T 1WI, T 2WI and diffusion weighted imaging (DWI) sequences. 3D data analysis module of the MaZda software was used to manually draw region of interest (ROIs) slice by slice on T 2WI and ADC images, and generate volume of interest (VOI) of the entire tumor. MaZda software was also used to extract texture features. The independent sample t test or Mann-Whitney U test were used to identify the texture features with statistically significant differences between low and high grade PCa groups. Lasso regression model was used to select the best combination of texture features for identifying low and high grade PCa, and then the model was built. The receiver operating characteristic (ROC) curve was used to evaluate the diagnostic performance of the model in both training cohort and test cohort. Results:The best texture feature combination selected by Lasso regression model were the S (1, 0, 0) correlation of T 2WI and the S (1, 0, 0) correlation, S (1, -1, 0) sum entropy and vertical-run length nonuniformity of ADC maps. The area under the ROC curve (AUC) of the model in training cohort was 0.823, and the sensitivity and specificity were 70.4% and 80.8%, respectively, which were better than the single texture feature. The AUC of the model in test cohort was 0.714, which was worse than training cohort. Conclusion:The texture analysis of T 2WI and ADC maps is valuable for the identification of low and high grade PCa.


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