1.Treatment Principles and Paradigm of Diabetic Microvascular Complications Responding Specifically to Traditional Chinese Medicine
Anzhu WANG ; Xing HANG ; Lili ZHANG ; Xiaorong ZHU ; Dantao PENG ; Ying FAN ; Min ZHANG ; Wenliang LYU ; Guoliang ZHANG ; Xiai WU ; Jia MI ; Jiaxing TIAN ; Wei ZHANG ; Han WANG ; Yuan XU ; .LI PINGPING ; Zhenyu WANG ; Ying ZHANG ; Dongmei SUN ; Yi HE ; Mei MO ; Xiaoxiao ZHANG ; Linhua ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(5):272-279
To explore the advantages of traditional Chinese medicine (TCM) and integrative TCM-Western medicine approaches in the treatment of diabetic microvascular complications (DMC), refine key pathophysiological insights and treatment principles, and promote academic innovation and strategic research planning in the prevention and treatment of DMC. The 38th session of the Expert Salon on Diseases Responding Specifically to Traditional Chinese Medicine, hosted by the China Association of Chinese Medicine, was held in Beijing, 2024. Experts in TCM, Western medicine, and interdisciplinary fields convened to conduct a systematic discussion on the pathogenesis, diagnostic and treatment challenges, and mechanism research related to DMC, ultimately forming a consensus on key directions. Four major research recommendations were proposed. The first is addressing clinical bottlenecks in the prevention and control of DMC by optimizing TCM-based evidence evaluation systems. The second is refining TCM core pathogenesis across DMC stages and establishing corresponding "disease-pattern-time" framework. The third is innovating mechanism research strategies to facilitate a shift from holistic regulation to targeted intervention in TCM. The fourth is advancing interdisciplinary collaboration to enhance the role of TCM in new drug development, research prioritization, and guideline formulation. TCM and integrative approaches offer distinct advantages in managing DMC. With a focus on the diseases responding specifically to TCM, strengthening evidence-based support and mechanism interpretation and promoting the integration of clinical care and research innovation will provide strong momentum for the modernization of TCM and the advancement of national health strategies.
2.Establishment of a mouse model of melasma using simple ultraviolet irradiation
Liying LI ; Jinghan SUN ; Cheng HUA ; Zhenyu CHEN ; Lele LYU ; Xian DU
Chinese Journal of Plastic Surgery 2024;40(4):444-454
Objective:To determine the optimal irradiation energy and frequency for the establishment of melasma mouse model using simple ultraviolet irradiation, and to provide guidance on animal strains and irradiation protocols for the successful establishment of melasma model.Methods:Animal models of melasma were established using BALB/c female mice and C57BL/6JNifdc female mice. BALB/c female mice were divided into 4 groups using a simple randomization method: A, B, C and G, with 5 mice in each group. C57BL/6JNifdc female mice were divided into 4 groups: D, E, F and H, with 5 mice in each group. All mice were irradiated with 8.428 mW/cm 2 of ultraviolet light. The irradiation time was 15 s (single irradiation energy of 0.13 J/cm 2) in groups A and D, 15 min (single irradiation energy of 7.59 J/cm 2) in groups B and E, and 30 min (single irradiation energy of 15.17 J/cm 2) in groups C and F. Each cycle consisted of 5 consecutive days of irradiation followed by 2 days of cessation, totaling 4 cycles of irradiation. Groups G and H were not irradiated. At the end of irradiation, all mice were kept under normal conditions. One week later, 3 mice from each group were selected for HE, Masson-Fontana, Masson, and immunohistochemical staining. Quantitative analysis was performed to measure the thickness of the acanthocyte layer, melanin granules, collagen percentage, and interleukin-1 (IL-1) levels. The remaining mice were kept for an additional week, depilated and photographed to observe the changes in coloration. Data were analyzed using SPSS 27.0 software, measurement data that did not conform to normal distribution were represented by M( Q1, Q3) and comparisons between groups were made using the Kruskal-Wallis rank sum test. Results:During the entire irradiation process, no visible discoloration was observed in the BALB/c female mice in all groups. In contrast, varying sizes of discoloration appeared in the C57BL/6JNifdc female mice in groups D, E, and F after irradiation in the second week. However, by the third week, the discoloration in group D gradually disappeared, while the discoloration in group E was more obvious than before. At the same time, group F exhibited significant discoloration, with some mice exhibited signs of skin peeling, burning and breakage on their backs. After the 4th week of irradiation, no new discoloration was formed in group D. The discoloration was more obvious in group E, and most mice in group F showed skin burn breakage. Two weeks after the completion of irradiation, there was no obvious discoloration on the dorsal skin of BALB/c female mice in all groups. In C57BL/6JNifdc female mice, group D showed no obvious discoloration, group E exhibited lighter discoloration compared to the 4th week post-irradiation, and group F had crusted skin at the burn sites with lighter discoloration than before. However, the discoloration in groups E and F was still obviously visible to the naked eye. HE staining showed that the difference in the thickness of the echinocyte layer was not statistically significant in groups A, B, C, and G ( H=1.08, P=0.782); whereas the difference was statistically significant in groups D, E, F and H ( H=12.85, P=0.005). The thickness of the echinocyte layer decreased gradually with the extension of the irradiation time. Additionally, there was a disruption in the arrangement of epidermal spindles in group F, and this situation was not observed in groups D and E. Masson-Fontana staining revealed no significant pigmentation in any of the BALB/c female mice. The difference in melanin granule counts between groups A, B, C, and G was not statistically significant ( H=7.77, P=0.051). In contrast, C57BL/6JNifdc female mice exhibited more noticeable pigmentation in the epidermis and dermis in groups E and F. The difference in melanin particle counts among groups D, E, F and H was statistically significant ( H=17.61, P<0.001), with melanin deposition increasing gradually with the duration of irradiation. Masson staining showed that the difference in collagen percentage between groups A, B, C, and G was not statistically significant ( H=7.26, P=0.064). However, significant disorganization of fibers and a loose structure were observed in groups E and F. The difference in collagen percentage between groups D, E, F, and H was statistically significant ( H=8.65, P=0.034). Immunohistochemical results showed that the difference in IL-1 expression levels between groups A, B, C, and G was statistically significant ( H=17.86, P<0.001); also between groups D, E, F, and H was statistically significant ( H=14.19, P=0.003), suggesting that ultraviolet irradiation stimulated an inflammatory response in the skin of mice. Conclusion:BALB/c female mice are not suitable for melasma models under the frequency and duration of irradiation in this experiment. C57BL/6JNifdc female mice are irradiated with a single irradiation energy dose of 7.59 J/cm 2 five days a week for 4 weeks, which can establish stable animal models of melasma with a specific level of pigmentation that persisted for at least 2 weeks.
3.Establishment of a mouse model of melasma using simple ultraviolet irradiation
Liying LI ; Jinghan SUN ; Cheng HUA ; Zhenyu CHEN ; Lele LYU ; Xian DU
Chinese Journal of Plastic Surgery 2024;40(4):444-454
Objective:To determine the optimal irradiation energy and frequency for the establishment of melasma mouse model using simple ultraviolet irradiation, and to provide guidance on animal strains and irradiation protocols for the successful establishment of melasma model.Methods:Animal models of melasma were established using BALB/c female mice and C57BL/6JNifdc female mice. BALB/c female mice were divided into 4 groups using a simple randomization method: A, B, C and G, with 5 mice in each group. C57BL/6JNifdc female mice were divided into 4 groups: D, E, F and H, with 5 mice in each group. All mice were irradiated with 8.428 mW/cm 2 of ultraviolet light. The irradiation time was 15 s (single irradiation energy of 0.13 J/cm 2) in groups A and D, 15 min (single irradiation energy of 7.59 J/cm 2) in groups B and E, and 30 min (single irradiation energy of 15.17 J/cm 2) in groups C and F. Each cycle consisted of 5 consecutive days of irradiation followed by 2 days of cessation, totaling 4 cycles of irradiation. Groups G and H were not irradiated. At the end of irradiation, all mice were kept under normal conditions. One week later, 3 mice from each group were selected for HE, Masson-Fontana, Masson, and immunohistochemical staining. Quantitative analysis was performed to measure the thickness of the acanthocyte layer, melanin granules, collagen percentage, and interleukin-1 (IL-1) levels. The remaining mice were kept for an additional week, depilated and photographed to observe the changes in coloration. Data were analyzed using SPSS 27.0 software, measurement data that did not conform to normal distribution were represented by M( Q1, Q3) and comparisons between groups were made using the Kruskal-Wallis rank sum test. Results:During the entire irradiation process, no visible discoloration was observed in the BALB/c female mice in all groups. In contrast, varying sizes of discoloration appeared in the C57BL/6JNifdc female mice in groups D, E, and F after irradiation in the second week. However, by the third week, the discoloration in group D gradually disappeared, while the discoloration in group E was more obvious than before. At the same time, group F exhibited significant discoloration, with some mice exhibited signs of skin peeling, burning and breakage on their backs. After the 4th week of irradiation, no new discoloration was formed in group D. The discoloration was more obvious in group E, and most mice in group F showed skin burn breakage. Two weeks after the completion of irradiation, there was no obvious discoloration on the dorsal skin of BALB/c female mice in all groups. In C57BL/6JNifdc female mice, group D showed no obvious discoloration, group E exhibited lighter discoloration compared to the 4th week post-irradiation, and group F had crusted skin at the burn sites with lighter discoloration than before. However, the discoloration in groups E and F was still obviously visible to the naked eye. HE staining showed that the difference in the thickness of the echinocyte layer was not statistically significant in groups A, B, C, and G ( H=1.08, P=0.782); whereas the difference was statistically significant in groups D, E, F and H ( H=12.85, P=0.005). The thickness of the echinocyte layer decreased gradually with the extension of the irradiation time. Additionally, there was a disruption in the arrangement of epidermal spindles in group F, and this situation was not observed in groups D and E. Masson-Fontana staining revealed no significant pigmentation in any of the BALB/c female mice. The difference in melanin granule counts between groups A, B, C, and G was not statistically significant ( H=7.77, P=0.051). In contrast, C57BL/6JNifdc female mice exhibited more noticeable pigmentation in the epidermis and dermis in groups E and F. The difference in melanin particle counts among groups D, E, F and H was statistically significant ( H=17.61, P<0.001), with melanin deposition increasing gradually with the duration of irradiation. Masson staining showed that the difference in collagen percentage between groups A, B, C, and G was not statistically significant ( H=7.26, P=0.064). However, significant disorganization of fibers and a loose structure were observed in groups E and F. The difference in collagen percentage between groups D, E, F, and H was statistically significant ( H=8.65, P=0.034). Immunohistochemical results showed that the difference in IL-1 expression levels between groups A, B, C, and G was statistically significant ( H=17.86, P<0.001); also between groups D, E, F, and H was statistically significant ( H=14.19, P=0.003), suggesting that ultraviolet irradiation stimulated an inflammatory response in the skin of mice. Conclusion:BALB/c female mice are not suitable for melasma models under the frequency and duration of irradiation in this experiment. C57BL/6JNifdc female mice are irradiated with a single irradiation energy dose of 7.59 J/cm 2 five days a week for 4 weeks, which can establish stable animal models of melasma with a specific level of pigmentation that persisted for at least 2 weeks.
4.Construction of a prediction model for postoperative prognosis in patients with resectable cholangiocarcinoma based on silence information regulator 2 expression
Wei WANG ; Wenbin JI ; Zhenyu LYU ; Wanliang SUN ; Yu SHAO ; Jing LIU ; Yan YANG
Journal of Zhejiang University. Medical sciences 2024;53(1):98-107
Objective:To develop a prediction model for postoperative prognosis in patients with cholangiocarcinoma(CCA)based on the expression of silence information regulator 2(SIRT2).Methods:The differential expression of SIRT2 between CCA and normal tissues was analyzed using TCGA and GEO databases.Gene set enrichment analysis(GSEA)was used to explore potential mechanisms of SIRT2 in CCA.The expression of SIRT2 protein in CCA tissues and normal tissues(including 44 pairs of specimens)was also detected by immunohistochemistry(IHC)in 89 resectable CCA patients who underwent surgical treatment in the First Affiliated Hospital of Bengbu Medical College between January 2016 and December 2021.The relationship between SIRT2 expression and clinicopathological characteristics and prognosis of CCA patients was analyzed.A survival prediction model for patients with resectable CCA was constructed with COX regression results,the calibration curve and the time-dependent receiver operating characteristic curve(ROC)were used to evaluate the performance of the constructed model,and the predictive power between this model and the American Joint Committee on Cancer(AJCC)/TNM staging system(8th edition)was compared.Results:SIRT2 mRNA was overexpressed in CCA tissues as shown in TCGA and GEO databases.IHC staining showed that SIRT2 protein expression in CCA tissues was significantly higher than that in adjacent non-tumor tissues.GSEA results showed that elevated SIRT2 expression may be involved in multiple metabolism-related signaling pathway,such as fatty acid metabolism,oxidative phosphorylation and amino acid metabolism.SIRT2 expression was related to serum triglycerides level,tumor size and lymph node metastasis(all P<0.05).The survival analysis results showed that patients with higher SIRT2 expression had a significantly lower overall survival(OS)than patients with lower SIRT2 expression(P<0.05).Univariate COX regression analysis suggested that pathological differentiation,clinical stage,postoperative treatment and SIRT2 expression level were associated with the prognosis of CCA patients(all P<0.05).Multivariate regression analysis confirmed that clinical stage and SIRT2 expression level were independent predictors of OS in postoperative CCA patients(both P<0.05).A nomogram based on SIRT2 for prediction of survival in postoperative CCA patients was constructed.The C-index of the model was 0.675,and the area under the time-dependent ROC curve(AUC)for predicting survival in the first,second,and third years was 0.879,0.778,and 0.953,respectively,which were superior to those of AJCC/TNM staging system(8th Edition).Conclusions:SIRT2 is highly expressed in CCA tissues,which is associated with poor prognosis in patients with resectable CCA.The nomogram developed based on SIRT2 may have better predictive power than the AJCC/TNM staging system(8th edition)in prediction of survival of postoperative CCA patients.
5.Construction of a prediction model for postoperative prognosis in patients with resectable cholangiocarcinoma based on silence information regulator 2 expression
Wei WANG ; Wenbin JI ; Zhenyu LYU ; Wanliang SUN ; Yu SHAO ; Jing LIU ; Yan YANG
Journal of Zhejiang University. Medical sciences 2024;53(1):98-107
Objective:To develop a prediction model for postoperative prognosis in patients with cholangiocarcinoma(CCA)based on the expression of silence information regulator 2(SIRT2).Methods:The differential expression of SIRT2 between CCA and normal tissues was analyzed using TCGA and GEO databases.Gene set enrichment analysis(GSEA)was used to explore potential mechanisms of SIRT2 in CCA.The expression of SIRT2 protein in CCA tissues and normal tissues(including 44 pairs of specimens)was also detected by immunohistochemistry(IHC)in 89 resectable CCA patients who underwent surgical treatment in the First Affiliated Hospital of Bengbu Medical College between January 2016 and December 2021.The relationship between SIRT2 expression and clinicopathological characteristics and prognosis of CCA patients was analyzed.A survival prediction model for patients with resectable CCA was constructed with COX regression results,the calibration curve and the time-dependent receiver operating characteristic curve(ROC)were used to evaluate the performance of the constructed model,and the predictive power between this model and the American Joint Committee on Cancer(AJCC)/TNM staging system(8th edition)was compared.Results:SIRT2 mRNA was overexpressed in CCA tissues as shown in TCGA and GEO databases.IHC staining showed that SIRT2 protein expression in CCA tissues was significantly higher than that in adjacent non-tumor tissues.GSEA results showed that elevated SIRT2 expression may be involved in multiple metabolism-related signaling pathway,such as fatty acid metabolism,oxidative phosphorylation and amino acid metabolism.SIRT2 expression was related to serum triglycerides level,tumor size and lymph node metastasis(all P<0.05).The survival analysis results showed that patients with higher SIRT2 expression had a significantly lower overall survival(OS)than patients with lower SIRT2 expression(P<0.05).Univariate COX regression analysis suggested that pathological differentiation,clinical stage,postoperative treatment and SIRT2 expression level were associated with the prognosis of CCA patients(all P<0.05).Multivariate regression analysis confirmed that clinical stage and SIRT2 expression level were independent predictors of OS in postoperative CCA patients(both P<0.05).A nomogram based on SIRT2 for prediction of survival in postoperative CCA patients was constructed.The C-index of the model was 0.675,and the area under the time-dependent ROC curve(AUC)for predicting survival in the first,second,and third years was 0.879,0.778,and 0.953,respectively,which were superior to those of AJCC/TNM staging system(8th Edition).Conclusions:SIRT2 is highly expressed in CCA tissues,which is associated with poor prognosis in patients with resectable CCA.The nomogram developed based on SIRT2 may have better predictive power than the AJCC/TNM staging system(8th edition)in prediction of survival of postoperative CCA patients.
6.Establishment of UPLC characteristic chromatogram of Pulsatilla chinensis and its application in origin differentiation and counterfeit identification
Guangming HE ; Rui LUO ; Heping ZENG ; Xiaoying LU ; Xiaolong YANG ; Weisheng LYU ; Yueyi LIANG ; Zhenyu LI ; Dongmei SUN ; Xiangdong CHEN
International Journal of Traditional Chinese Medicine 2024;46(6):743-749
Objective:To establish ultra performance liquid chromatography (UPLC) characteristic chromatogram of Pulsatilla chinensis; To provide reference for the origin identification and quality control of Pulsatilla chinensis. Methods:UPLC Method was adopted. The determination was performed on a column of Agilent SB C18 (2.1 mm×100 mm, 1.8 μm) . The mobile phase was acetonitrile-methanol (2:1) -0.1% phosphoric acid solution by fradient elution at a flow rate of 0.30ml/min. The column temperature was 30 ℃. The detection wavelength was 215 nm. The injection volume was 2 μl. The common counterfeit products and medicinal herbs of Pulsatilla chinensis from different areas were evaluated by comparison of characteristic chromatogram, principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA). Results:There were 9 characteristic peaks in the characteristic chromatogram of Pulsatilla chinensis, and 8 common peaks were identified by high resolution mass spectrometry and comparison of reference materials. Through PCA analysis, it was possible to clearly distinguish the medicinal herbs of Pulsatilla chinensis from different areas. Combined with OPLS-DA analysis, it was found that peak 2, peak 3, peak 6 were the main markers of Pulsatilla chinensis from different producing areas. Conclusion:The established method has good specificity, repeatability and durability, and it can effectively distinguish the common counterfeits of Pulsatilla chinensis, and provide the basis of quality control and selection of origin for Pulsatilla chinensis.
7.Study on UPLC fingerprint of Mume flos at different flowering stages based on chemometrics analysis
Shuang HUANG ; Yueyi LIANG ; Jie YANG ; Weisheng LYU ; Xiaoying LU ; Guangming HE ; Zhipeng CHEN ; Xuxuan HOU ; Tianrui XIA ; Zhenyu LI ; Congyou DENG ; Xiangdong CHEN ; Dongmei SUN
International Journal of Traditional Chinese Medicine 2024;46(7):898-904
Objective:To establish the ultra high performance liquid chromatography (UPLC) fingerprints of Mume flos at different flowering stages; To provide reference for the quality research of Mume flos.Methods:The fingerprints of Mume flos were established by UPLC method, and the common peaks were identified by high performance liquid chromatography high resolution mass spectrometry (LC-MS). Chemometrics analysis was carried out with the fingerprints' common peak area of plum blossom at different flowering stages as a variable. Semiquantitative analysis of changes in flavonoids and phenolic acids in Mume flos at different flowering stages was conduct using peak area calculation method.Results:Totally 31 common peaks were identified in the fingerprints of plum blossom medicinal materials at different flowering stages and 9 components were identified. Clustering analysis (HCA) and principal component analysis (PCA) both classified plum blossom medicinal herbs at different flowering stages into three categories. Among them, there were significant differences between the groups at the bud stage, blooming period, and final flowering period, while the differences between the groups at blooming period and final flowering period were relatively small. The orthogonal partial least squares discriminant analysis (OPLS-DA) screened 16 different components with VIP>1.0. The contents of phenolic acids in different flowering stages were as follows: bud stage>blooming period>final flowering period, while the contents of flavonoids were as follows: blooming period>final flowering period>bud stage.Conclusions:This method is simple and reliable, and can provide reference for the quality evaluation of plum blossom medicinal materials at different flowering stages.
8.Construction of a prediction model for postoperative prognosis in patients with resectable cholangiocarcinoma based on silence information regulator 2 expression
Wei WANG ; Wenbin JI ; Zhenyu LYU ; Wanliang SUN ; Yu SHAO ; Jing LIU ; Yan YANG
Journal of Zhejiang University. Medical sciences 2024;53(1):98-107
Objective:To develop a prediction model for postoperative prognosis in patients with cholangiocarcinoma(CCA)based on the expression of silence information regulator 2(SIRT2).Methods:The differential expression of SIRT2 between CCA and normal tissues was analyzed using TCGA and GEO databases.Gene set enrichment analysis(GSEA)was used to explore potential mechanisms of SIRT2 in CCA.The expression of SIRT2 protein in CCA tissues and normal tissues(including 44 pairs of specimens)was also detected by immunohistochemistry(IHC)in 89 resectable CCA patients who underwent surgical treatment in the First Affiliated Hospital of Bengbu Medical College between January 2016 and December 2021.The relationship between SIRT2 expression and clinicopathological characteristics and prognosis of CCA patients was analyzed.A survival prediction model for patients with resectable CCA was constructed with COX regression results,the calibration curve and the time-dependent receiver operating characteristic curve(ROC)were used to evaluate the performance of the constructed model,and the predictive power between this model and the American Joint Committee on Cancer(AJCC)/TNM staging system(8th edition)was compared.Results:SIRT2 mRNA was overexpressed in CCA tissues as shown in TCGA and GEO databases.IHC staining showed that SIRT2 protein expression in CCA tissues was significantly higher than that in adjacent non-tumor tissues.GSEA results showed that elevated SIRT2 expression may be involved in multiple metabolism-related signaling pathway,such as fatty acid metabolism,oxidative phosphorylation and amino acid metabolism.SIRT2 expression was related to serum triglycerides level,tumor size and lymph node metastasis(all P<0.05).The survival analysis results showed that patients with higher SIRT2 expression had a significantly lower overall survival(OS)than patients with lower SIRT2 expression(P<0.05).Univariate COX regression analysis suggested that pathological differentiation,clinical stage,postoperative treatment and SIRT2 expression level were associated with the prognosis of CCA patients(all P<0.05).Multivariate regression analysis confirmed that clinical stage and SIRT2 expression level were independent predictors of OS in postoperative CCA patients(both P<0.05).A nomogram based on SIRT2 for prediction of survival in postoperative CCA patients was constructed.The C-index of the model was 0.675,and the area under the time-dependent ROC curve(AUC)for predicting survival in the first,second,and third years was 0.879,0.778,and 0.953,respectively,which were superior to those of AJCC/TNM staging system(8th Edition).Conclusions:SIRT2 is highly expressed in CCA tissues,which is associated with poor prognosis in patients with resectable CCA.The nomogram developed based on SIRT2 may have better predictive power than the AJCC/TNM staging system(8th edition)in prediction of survival of postoperative CCA patients.
9.Construction of a prediction model for postoperative prognosis in patients with resectable cholangiocarcinoma based on silence information regulator 2 expression
Wei WANG ; Wenbin JI ; Zhenyu LYU ; Wanliang SUN ; Yu SHAO ; Jing LIU ; Yan YANG
Journal of Zhejiang University. Medical sciences 2024;53(1):98-107
Objective:To develop a prediction model for postoperative prognosis in patients with cholangiocarcinoma(CCA)based on the expression of silence information regulator 2(SIRT2).Methods:The differential expression of SIRT2 between CCA and normal tissues was analyzed using TCGA and GEO databases.Gene set enrichment analysis(GSEA)was used to explore potential mechanisms of SIRT2 in CCA.The expression of SIRT2 protein in CCA tissues and normal tissues(including 44 pairs of specimens)was also detected by immunohistochemistry(IHC)in 89 resectable CCA patients who underwent surgical treatment in the First Affiliated Hospital of Bengbu Medical College between January 2016 and December 2021.The relationship between SIRT2 expression and clinicopathological characteristics and prognosis of CCA patients was analyzed.A survival prediction model for patients with resectable CCA was constructed with COX regression results,the calibration curve and the time-dependent receiver operating characteristic curve(ROC)were used to evaluate the performance of the constructed model,and the predictive power between this model and the American Joint Committee on Cancer(AJCC)/TNM staging system(8th edition)was compared.Results:SIRT2 mRNA was overexpressed in CCA tissues as shown in TCGA and GEO databases.IHC staining showed that SIRT2 protein expression in CCA tissues was significantly higher than that in adjacent non-tumor tissues.GSEA results showed that elevated SIRT2 expression may be involved in multiple metabolism-related signaling pathway,such as fatty acid metabolism,oxidative phosphorylation and amino acid metabolism.SIRT2 expression was related to serum triglycerides level,tumor size and lymph node metastasis(all P<0.05).The survival analysis results showed that patients with higher SIRT2 expression had a significantly lower overall survival(OS)than patients with lower SIRT2 expression(P<0.05).Univariate COX regression analysis suggested that pathological differentiation,clinical stage,postoperative treatment and SIRT2 expression level were associated with the prognosis of CCA patients(all P<0.05).Multivariate regression analysis confirmed that clinical stage and SIRT2 expression level were independent predictors of OS in postoperative CCA patients(both P<0.05).A nomogram based on SIRT2 for prediction of survival in postoperative CCA patients was constructed.The C-index of the model was 0.675,and the area under the time-dependent ROC curve(AUC)for predicting survival in the first,second,and third years was 0.879,0.778,and 0.953,respectively,which were superior to those of AJCC/TNM staging system(8th Edition).Conclusions:SIRT2 is highly expressed in CCA tissues,which is associated with poor prognosis in patients with resectable CCA.The nomogram developed based on SIRT2 may have better predictive power than the AJCC/TNM staging system(8th edition)in prediction of survival of postoperative CCA patients.
10.Construction of a prediction model for postoperative prognosis in patients with resectable cholangiocarcinoma based on silence information regulator 2 expression
Wei WANG ; Wenbin JI ; Zhenyu LYU ; Wanliang SUN ; Yu SHAO ; Jing LIU ; Yan YANG
Journal of Zhejiang University. Medical sciences 2024;53(1):98-107
Objective:To develop a prediction model for postoperative prognosis in patients with cholangiocarcinoma(CCA)based on the expression of silence information regulator 2(SIRT2).Methods:The differential expression of SIRT2 between CCA and normal tissues was analyzed using TCGA and GEO databases.Gene set enrichment analysis(GSEA)was used to explore potential mechanisms of SIRT2 in CCA.The expression of SIRT2 protein in CCA tissues and normal tissues(including 44 pairs of specimens)was also detected by immunohistochemistry(IHC)in 89 resectable CCA patients who underwent surgical treatment in the First Affiliated Hospital of Bengbu Medical College between January 2016 and December 2021.The relationship between SIRT2 expression and clinicopathological characteristics and prognosis of CCA patients was analyzed.A survival prediction model for patients with resectable CCA was constructed with COX regression results,the calibration curve and the time-dependent receiver operating characteristic curve(ROC)were used to evaluate the performance of the constructed model,and the predictive power between this model and the American Joint Committee on Cancer(AJCC)/TNM staging system(8th edition)was compared.Results:SIRT2 mRNA was overexpressed in CCA tissues as shown in TCGA and GEO databases.IHC staining showed that SIRT2 protein expression in CCA tissues was significantly higher than that in adjacent non-tumor tissues.GSEA results showed that elevated SIRT2 expression may be involved in multiple metabolism-related signaling pathway,such as fatty acid metabolism,oxidative phosphorylation and amino acid metabolism.SIRT2 expression was related to serum triglycerides level,tumor size and lymph node metastasis(all P<0.05).The survival analysis results showed that patients with higher SIRT2 expression had a significantly lower overall survival(OS)than patients with lower SIRT2 expression(P<0.05).Univariate COX regression analysis suggested that pathological differentiation,clinical stage,postoperative treatment and SIRT2 expression level were associated with the prognosis of CCA patients(all P<0.05).Multivariate regression analysis confirmed that clinical stage and SIRT2 expression level were independent predictors of OS in postoperative CCA patients(both P<0.05).A nomogram based on SIRT2 for prediction of survival in postoperative CCA patients was constructed.The C-index of the model was 0.675,and the area under the time-dependent ROC curve(AUC)for predicting survival in the first,second,and third years was 0.879,0.778,and 0.953,respectively,which were superior to those of AJCC/TNM staging system(8th Edition).Conclusions:SIRT2 is highly expressed in CCA tissues,which is associated with poor prognosis in patients with resectable CCA.The nomogram developed based on SIRT2 may have better predictive power than the AJCC/TNM staging system(8th edition)in prediction of survival of postoperative CCA patients.

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