1.Expression of MYPT1, CPI17, and PP1C in bile duct epithelium of children with pancreaticobiliary maljunction
Jungang ZHAO ; Chenxiang HAO ; Wanliang GUO ; Shungen HUANG ; Jian WANG
Chinese Journal of Hepatobiliary Surgery 2025;31(11):851-855
Objective:To analyze the expression of myosin light chain phosphatase target subunit-1 (MYPT1), protein kinase C-activated phosphatase inhibitor 17 (CPI17), and protein phosphatase 1 catalytic subunit (PP1C) in the biliary epithelium of children with pancreaticobiliary maljunction (PBM).Methods:A total of 22 children with PBM and biliary dilatation from the Children's Hospital of Soochow University from March 2015 to December 2017 were collected as the PBM group ( n=22), including 8 males and 14 females, with an age of 4.2 (2.8, 6.3) years. 21 patients who underwent cholecystectomy for gallstones at the First Affiliated Hospital of Soochow University were collected as the control group ( n=21), including 9 males and 12 females, with an age of 32.2 (23.8, 48.3) years. The expression of tight junction-related proteins pMLC, CPI17, PP1C, and MYPT1 in the PBM group and the control group was analyzed by immunohistochemical staining combined score. Western blot was used to detect the protein expression of MYPT1, PP1C, MLC, pCPI17, CPI17, and pMYPT1 in the PBM group and the control group. Results:The immunohistochemical staining combined score showed that the expression of pMLC protein in the PBM group was higher than that in the control group, while the expression of MYPT1, CPI17, and PP1C proteins was lower than that in the control group, and the differences were statistically significant (all P<0.05). The Western blot detection results showed that the expression values of pMYPT1 (Thr696), pCPI17 (Thr38), MLC, CPI17, MYPT1, and PP1C proteins in the PBM group were (1.187±0.140), (0.455±0.038), (0.181±0.013), (0.246±0.016), (0.459±0.054), and (0.202±0.032), respectively, while those in the control group were (0.238±0.048), (0.144±0.025), (3.268±0.278), (1.462±0.214), (1.138±0.189), and (0.853±0.120), respectively. The expression of MLC, CPI17, MYPT1, and PP1C proteins in the PBM group was lower than that in the control group, while the expression of pMYPT1 and pCPI17 proteins was higher than that in the control group, and the differences were statistically significant (all P<0.05). Conclusion:The increased expression of pMYPT1 and pCPI17 and the decreased expression of PP1C in children with PBM may be related to the disruption of tight junctions in the biliary epithelium of these children.
2.Expression of MYPT1, CPI17, and PP1C in bile duct epithelium of children with pancreaticobiliary maljunction
Jungang ZHAO ; Chenxiang HAO ; Wanliang GUO ; Shungen HUANG ; Jian WANG
Chinese Journal of Hepatobiliary Surgery 2025;31(11):851-855
Objective:To analyze the expression of myosin light chain phosphatase target subunit-1 (MYPT1), protein kinase C-activated phosphatase inhibitor 17 (CPI17), and protein phosphatase 1 catalytic subunit (PP1C) in the biliary epithelium of children with pancreaticobiliary maljunction (PBM).Methods:A total of 22 children with PBM and biliary dilatation from the Children's Hospital of Soochow University from March 2015 to December 2017 were collected as the PBM group ( n=22), including 8 males and 14 females, with an age of 4.2 (2.8, 6.3) years. 21 patients who underwent cholecystectomy for gallstones at the First Affiliated Hospital of Soochow University were collected as the control group ( n=21), including 9 males and 12 females, with an age of 32.2 (23.8, 48.3) years. The expression of tight junction-related proteins pMLC, CPI17, PP1C, and MYPT1 in the PBM group and the control group was analyzed by immunohistochemical staining combined score. Western blot was used to detect the protein expression of MYPT1, PP1C, MLC, pCPI17, CPI17, and pMYPT1 in the PBM group and the control group. Results:The immunohistochemical staining combined score showed that the expression of pMLC protein in the PBM group was higher than that in the control group, while the expression of MYPT1, CPI17, and PP1C proteins was lower than that in the control group, and the differences were statistically significant (all P<0.05). The Western blot detection results showed that the expression values of pMYPT1 (Thr696), pCPI17 (Thr38), MLC, CPI17, MYPT1, and PP1C proteins in the PBM group were (1.187±0.140), (0.455±0.038), (0.181±0.013), (0.246±0.016), (0.459±0.054), and (0.202±0.032), respectively, while those in the control group were (0.238±0.048), (0.144±0.025), (3.268±0.278), (1.462±0.214), (1.138±0.189), and (0.853±0.120), respectively. The expression of MLC, CPI17, MYPT1, and PP1C proteins in the PBM group was lower than that in the control group, while the expression of pMYPT1 and pCPI17 proteins was higher than that in the control group, and the differences were statistically significant (all P<0.05). Conclusion:The increased expression of pMYPT1 and pCPI17 and the decreased expression of PP1C in children with PBM may be related to the disruption of tight junctions in the biliary epithelium of these children.
3.Comparison of Effect of Different Microbial Agents on Quality of Lycii Fructus Based on Multiple Index Components
Jia HE ; Youyuan LU ; Mingxia XIE ; Haixia SUN ; Tao WANG ; Jiahui XUE ; Wanliang HAO ; Hanqing WANG
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(24):177-184
ObjectiveTo reveal the effects of different microbial agents on quality of Lycii Fructus by comparing the differences in the contents of multiple types of chemical components in Lycii Fructus after the application of different microbial agents. MethodTaking Ningqi No. 7 as experimental material, four microbial agents, namely Peiyuan combined with Xinterui(TP group), Trichoderma harzianum combined with Bacillus subtilis(BW group), Genwuyou(MT group) and Junyiduo(JYD group), were applied, and no microbial agents was used as the blank group(CK group). Then the contents of total phenolics, total flavonoids, saccharides, amino acids, nucleosides and bases, betaine and other components in Lycii Fructus were determined by ultraviolet spectrophotometry(UV), high performance liquid chromatography(HPLC) and ultra-high performance liquid chromatography-mass spectrometry(UHPLC-MS/MS), and the methods such as multiple comparisons, principal component analysis(PCA) and partial least squares-discriminant analysis(PLS-DA) were used to analyze the effect of different microbial agents on the quality of Lycii Fructus. ResultMicrobial agents had different effects on chemical components of Lycii Fructus. The content of total phenolics was the highest in the TP group, and it varied significantly from the CK group(P<0.05). The total flavonoid content was the highest in the BW group, followed by the TP group. Both polysaccharide and alduronic acid contents were the highest in the JYD group. Betaine content in the TP and BW groups were significantly higher than that in the CK group(P<0.05). For the determined 23 kinds of amino acids, most of them were the lowest in the JYD group, and the highest in the MT group, while the nucleoside bases were higher in the MT and BW groups. It indicated that Lycii Fructus from different treatment groups could be distinguished clearly based on the determined 45 chemical components. The result of PLS-DA showed that the major differential components in each group were polysaccharides, glucose, fructose, betaine, alduronic acid, asparagine, sucrose, threonine, total flavonoids, alanine and total phenolics. The results of PCA composite scores based on the main differential components showed that composite scores of chemical components in each group were BW group>TP group>MT group>CK group>JYD group. ConclusionThe application of microbial agents of BW, TP and MT can promote the quality improvement of Lycii Fructus, and the application of JYD can promote the accumulation of polysaccharides and alduronic acid to a certain extent, but the overall effect on the quality of Lycii Fructus is not clear. This study lays the foundation for the green and healthy development of Lycii Fructus industry.

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