1.Mechanisms of Gegen Qinlian Tang-containing Serum in Improving 5-FU Sensitivity by Inhibiting Glycolysis in Colorectal Cancer Cells Based on CDK16/MYC Pathway
Rong CAI ; Shang WANG ; Fuqing CHENG ; Yanping ZHOU ; Zuowei HU ; Yunhai LI
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(8):1-9
ObjectiveTo explore the molecular mechanisms by which serum containing Gegen Qinlian Tang (GQT) inhibits glycolysis and enhances chemotherapy sensitivity in 5-fluorouracil (5-FU)-resistant colorectal cancer (CRC) cells based on the cyclin-dependent kinase 16 (CDK16)/MYC proto-oncogene (MYC) pathway. MethodsHCT-116/5-FU cells were treated with different concentrations (5%, 10%, 20%, 30%) of GQT-containing serum. Cell viability and 5-FU sensitivity were assessed using the cell counting kit-8 (CCK-8) assay, and the experimental concentrations of 5-FU and GQT for subsequent experiments were determined. Cell proliferation and apoptosis under individual 5-FU, GQT, and combined 5-FU + GQT treatments were evaluated using 5-ethynyl-2′-deoxyuridine (EDU) staining and annexin V-FITC/PI double staining, respectively. Glucose consumption, adenosine triphosphate (ATP) production, and lactate levels were measured by colorimetric assays. Expression levels of glycolysis-related proteins, CDK16, MYC, and phosphorylated MYC were detected by Western blot. Co-immunoprecipitation (CoIP) was used to examine the protein interaction between CDK16 and MYC, and cycloheximide (CHX) treatment was applied to assess the effect of CDK16 overexpression on MYC protein stability. ResultsCCK-8 assays showed that 2.5 mg·L-1 5-FU significantly inhibited HCT-116 cell viability in a dose-dependent manner. In HCT-116/5-FU cells, significant inhibition was observed only at 5 mg·L-1 5-FU (P<0.05), which was used for model establishment. Compared with 5-FU alone, addition of 5% GQT-containing serum significantly suppressed HCT-116/5-FU cell viability (P<0.05), with stronger inhibition at higher serum concentrations. Thus, 5% GQT-containing serum was used in subsequent experiments. Compared with the control group, 5-FU, GQT, and 5-FU + GQT treatments all significantly reduced cell proliferation (P<0.05) and increased apoptosis (P<0.01). The 5-FU + GQT combination showed superior inhibition of proliferation compared with 5-FU or GQT alone (P<0.01), accompanied by more pronounced reductions in glucose consumption, ATP production, and lactate generation (P<0.01). Additionally, compared with control, 5-FU, and GQT groups, the 5-FU + GQT group exhibited stronger suppression of MYC and its phosphorylated forms (P<0.01) and greater inhibition of glycolytic enzymes, including hexokinase 2 (HK2), 3-phosphoinositide-dependent protein kinase 1 (PDK1), lactate dehydrogenase A (LDHA), and pyruvate kinase M2 (PKM2) (P<0.01). CDK16, MYC, and MYC phosphorylation expression levels were significantly downregulated in the 5-FU + GQT group compared with the 5-FU group (all P<0.01). MYC protein stability decreased in a time-dependent manner in the 5-FU + GQT group (P<0.05), which was rescued by CDK16 overexpression (P<0.05). ConclusionGQT significantly enhances the sensitivity of HCT-116/5-FU cells to 5-FU, potentially by inhibiting CDK16 and thereby reducing MYC-mediated glycolysis.
2.Mechanisms of Gegen Qinlian Tang-containing Serum in Improving 5-FU Sensitivity by Inhibiting Glycolysis in Colorectal Cancer Cells Based on CDK16/MYC Pathway
Rong CAI ; Shang WANG ; Fuqing CHENG ; Yanping ZHOU ; Zuowei HU ; Yunhai LI
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(8):1-9
ObjectiveTo explore the molecular mechanisms by which serum containing Gegen Qinlian Tang (GQT) inhibits glycolysis and enhances chemotherapy sensitivity in 5-fluorouracil (5-FU)-resistant colorectal cancer (CRC) cells based on the cyclin-dependent kinase 16 (CDK16)/MYC proto-oncogene (MYC) pathway. MethodsHCT-116/5-FU cells were treated with different concentrations (5%, 10%, 20%, 30%) of GQT-containing serum. Cell viability and 5-FU sensitivity were assessed using the cell counting kit-8 (CCK-8) assay, and the experimental concentrations of 5-FU and GQT for subsequent experiments were determined. Cell proliferation and apoptosis under individual 5-FU, GQT, and combined 5-FU + GQT treatments were evaluated using 5-ethynyl-2′-deoxyuridine (EDU) staining and annexin V-FITC/PI double staining, respectively. Glucose consumption, adenosine triphosphate (ATP) production, and lactate levels were measured by colorimetric assays. Expression levels of glycolysis-related proteins, CDK16, MYC, and phosphorylated MYC were detected by Western blot. Co-immunoprecipitation (CoIP) was used to examine the protein interaction between CDK16 and MYC, and cycloheximide (CHX) treatment was applied to assess the effect of CDK16 overexpression on MYC protein stability. ResultsCCK-8 assays showed that 2.5 mg·L-1 5-FU significantly inhibited HCT-116 cell viability in a dose-dependent manner. In HCT-116/5-FU cells, significant inhibition was observed only at 5 mg·L-1 5-FU (P<0.05), which was used for model establishment. Compared with 5-FU alone, addition of 5% GQT-containing serum significantly suppressed HCT-116/5-FU cell viability (P<0.05), with stronger inhibition at higher serum concentrations. Thus, 5% GQT-containing serum was used in subsequent experiments. Compared with the control group, 5-FU, GQT, and 5-FU + GQT treatments all significantly reduced cell proliferation (P<0.05) and increased apoptosis (P<0.01). The 5-FU + GQT combination showed superior inhibition of proliferation compared with 5-FU or GQT alone (P<0.01), accompanied by more pronounced reductions in glucose consumption, ATP production, and lactate generation (P<0.01). Additionally, compared with control, 5-FU, and GQT groups, the 5-FU + GQT group exhibited stronger suppression of MYC and its phosphorylated forms (P<0.01) and greater inhibition of glycolytic enzymes, including hexokinase 2 (HK2), 3-phosphoinositide-dependent protein kinase 1 (PDK1), lactate dehydrogenase A (LDHA), and pyruvate kinase M2 (PKM2) (P<0.01). CDK16, MYC, and MYC phosphorylation expression levels were significantly downregulated in the 5-FU + GQT group compared with the 5-FU group (all P<0.01). MYC protein stability decreased in a time-dependent manner in the 5-FU + GQT group (P<0.05), which was rescued by CDK16 overexpression (P<0.05). ConclusionGQT significantly enhances the sensitivity of HCT-116/5-FU cells to 5-FU, potentially by inhibiting CDK16 and thereby reducing MYC-mediated glycolysis.
3.Study on Correlation Between Traditional Chinese Medicine Syndrome Elements and Risk Factors in Patients with Type Ⅱ Cardio-renal Syndrome
Rong YU ; Hanchao HUANG ; Huan DENG ; Wenli DING ; Yanping ZENG
Journal of Guangzhou University of Traditional Chinese Medicine 2025;42(7):1568-1573
Objective To investigate the correlation between traditional Chinese medicine(TCM)syndrome elements and risk factors in patients with type Ⅱ cardio-renal syndrome(CRS).Methods A retrospective analysis was conducted on the medical records of 116 patients with type Ⅱ CRS.The distribution of TCM syndrome elements was analyzed,and the correlation between the syndrome elements and risk factors of gender,age,smoking history,alcohol consumption history,coronary heart disease,hypertension,diabetes,and anemia were evaluated.Results(1)The analysis of syndrome elements of 116 patients with type Ⅱ CRS showed that the main disease-location elements were heart(96 cases,82.76%),lung(89 cases,76.72%),kidney(81 cases,69.83%),spleen(71 cases,61.21%),and liver(25 cases,21.56%);the main disease-nature elements were qi deficiency(113 cases,97.41%),blood deficiency(96 cases,82.76%),yang deficiency(95 cases,81.89%),phlegm(94 cases,81.03%),yin deficiency(90 cases,77.59%),dampness(61 cases,52.59%),water retention(55 cases,47.41%),and blood stasis(47 cases,40.52%).(2)The analysis of age groups of 116 patients with type Ⅱ CRS showed that the majority of the patients were middle-aged and elderly individuals over 60 years old,and the age groups covered<60 years old(4 cases,3.45%),61-70 years old(10 cases,8.62%),71-80 years old(28 cases,24.14%),81-90 years old(64 cases,55.17%),and>90 years old(10 cases,8.62%).(3)The analysis of gender of 116 patients with type Ⅱ CRS showed that 46 cases(39.66%)were male,and 70 cases(60.34%)were female,the female outnumbering the male.(4)The exploration of correlation between TCM syndrome elements and risk factors with Logistic regression analysis showed that blood deficiency was positively correlated with alcohol consumption,hypertension,and anemia(P<0.05 or P<0.01),yang deficiency was positively correlated with anemia(P<0.05),yin deficiency was positively correlated with smoking and diabetes(P<0.05),dampness was positively correlated with smoking and anemia(P<0.01),water retention was positively correlated with gender and protein abnormalities(P<0.05 or P<0.01),and blood stasis was positively correlated with hypertension(P<0.05).Conclusion The illness of type Ⅱ CRS is located in the heart,involving the five organs,and is closely related to the lungs and kidneys.The general pathogenesis of type Ⅱ CRS is characterized by being deficiency of qi and yang in the root cause,and having the symptom manifestations of phlegm and blood stasis.In type Ⅱ CRS patients,men are more likely to suffer water retention,smokers are more likely to suffer yin deficiency and dampness,drinkers are more likely to suffer blood deficiency,individuals with hypertension are more likely to suffer blood deficiency and blood stasis,individuals with diabetes are more likely to suffer yin deficiency,individuals with anemia are more likely to suffer blood deficiency,yang deficiency and dampness,and individuals with protein abnormalities are more likely to suffer water retention.
4.miR-21-5p Targetes STAT3 Reduce the OGD/R-induced Neuronal Injury
Yanping LI ; Qingyun LI ; Rong HUO ; Xiaolin DONG ; Hongmei LI ; Huan WEI ; Yi ZENG
Journal of Kunming Medical University 2025;46(5):12-20
Objective To investigate the potential mechanism of miR-21-5p in alleviating cerebral ischemia-reperfusion injury by targeting STAT3.Methods The HT22 cells were induced by OGD/R to construct a cell model of cerebral ischemia reperfusion injury.The expression of miR-21-5p was detected by RT-qPCR.The CCK-8 assay,TUNEL staining and flow cytometry were respectively used to detect the cell viability and apoptosis.ELISA assay was used to determine the contents of inflammatory factors IL-6,IL-10 and TNF-α in the cell supernatant.Western blot was used to detect the expression levels of p-STAT3/STAT3,Cleaved-Caspase-3,Bax and Bcl-2 proteins.The TargetScan database was used to predict the binding sites of miR-21-5p and STAT3.The dual-luciferase reporter gene assay was used to verify the targeting relationship between miR-21-5p and STAT3.Results The relative expression level of miR-21-5p was down-regulated in HT22 cells which induced by OGD/R(P<0.001).The cell viability(P<0.0001)was decreased and the apoptosis rate(P<0.001)was increased in OGD/R induced-HT22 cells.The contents of pro-inflammatory factors IL-6(P<0.001)and TNF-α(P<0.001)was increased,while the content of anti-inflammatory factor IL-10(P<0.001)decreased.After transfection with miR-21-5p mimic,cell viability was enhanced,apoptosis rate was reduced and neuroinflammation was inhibited.MiR-21-5p could target and bind to STAT3.After miR-21-5p inhibitor transfection,cell viability decreased,apoptosis was promoted,and neuroinflammation occurred;STAT3 inhibitor Stattic could reverse the effect of miR-21-5p inhibitor.Conclusion MiR-21-5p could specifically bind to STAT3 and reduce the neuroinflammation and apoptosis of OGD/R induced-HT22 cells.
5.Development, reliability, and validity of a treatment-related quality of life scale for Chinese patients with multiple myeloma
Chunyan SUN ; Zhen CAI ; Bing CHEN ; Lijuan CHEN ; Wenming CHEN ; Kaiyang DING ; Juan DU ; Rong FU ; Chengcheng FU ; Da GAO ; Guangxun GAO ; Yanjuan HE ; Jian HOU ; Ming JIANG ; Fei LI ; Jian LI ; Juan LI ; Zhenyu LI ; Aijun LIAO ; Jing LIU ; Jun LUO ; Jianmin LUO ; Yanping MA ; Jianqing MI ; Ting NIU ; Hongling PENG ; Yongping SONG ; Luqun WANG ; Rong ZHAN ; Xi ZHANG ; Yu HU
Chinese Journal of Hematology 2025;46(8):713-721
Objective:To develop a treatment-related quality of life scale for Chinese patients with multiple myeloma (MM) and to test its reliability and validity.Methods:The initial scale was constructed through a literature search, Delphi expert correspondence, and cognitive testing. This study conducted a preliminary survey of 379 patients with MM and a formal survey of 865 patients from the hematology departments of 155 hospitals nationwide from February 2024 to March 2024. The final scale was obtained after conducting item analysis and reliability and validity tests on the initial scale.Results:The constructed scale contains 36 items covering six domains: physiological, psychological, social, treatment side effects, general health, and others. In the preliminary survey, the Cronbach’s alpha coefficient of each item ranged from 0.597 to 0.939, and the test-retest reliability was 0.747 ( P<0.001). Exploratory factor analysis extracted eight common factors with a cumulative variance contribution of 60.058%. In the formal survey, the Cronbach’s alpha coefficient of each item ranged from 0.484 to 0.930, and the test-retest reliability was 0.835 ( P<0.001). Confirmatory factor analysis revealed a comparative fit index of 0.750, a root-mean-square error of approximation of 0.090, and a root-mean-square residual of 0.067. Conclusion:The treatment-related quality of life scale for Chinese patients with MM designed in this study exhibited good reliability and validity, reflecting the impact of treatment on the quality of life of patients. This scale can provide a reference to clinicians for assessing the disease status of patients.
6.A Study on the Risk of Cardiovascular Health and Pelvic Inflammatory Disease Based on the 2013-2018 National Health and Nutrition Examination Survey Data
Shuangshuang XIE ; Yangxue YIN ; Liming YUAN ; Lingyun LIAO ; Qin XU ; Yanping ZHANG ; Rong ZHOU
Journal of Practical Obstetrics and Gynecology 2025;41(5):400-405
Objective:To analyze the relationship between Cardiovascular Health(CVH)and Pelvic Inflamma-tory Disease(PID)using data from the National Health and Nutrition Examination Surveys(NHANES).Meth-ods:Participants from the NHANES database were extracted based on the inclusion of PID,LE8 scores,and mul-tiple potential confounding factors,excluding those with missing variables.Multivariate Logistic regression analysis was conducted using R to assess the relationship between the CVH and the risk of PID.Additionally,subgroup a-nalysis and interaction tests were performed for age,race,poverty,marital status,education,BMI,menstrual regu-larity,and pregnancy history.Results:A total of 3934 female participants were included,with an average age of 39.95±11.42 years.Multivariable Logistic regression analysis showed that after adjusting for age,race,BMI,edu-cation level,marital status,household income,menstrual cycle,and pregnancy history,the risk of PID gradually decreased with an increase in CVH assessment metric LE8 scores(OR 0.98,95%CI 0.97-0.99,P<0.001).Stratified analysis indicated significant interactions between age,race,and marital status in the association be-tween CVH and PID(interaction P<0.05 for all),while household income level,education level,BMI,menstrual regularity,and pregnancy history showed no significant interactions with the CVH and PID association(P>0.05).Conclusions:There is a significant negative relationship between CVH and PID in adult women in the United states,suggesting that better cardiovascular health may help reduce the risk of PID.
7.A Study on the Risk of Cardiovascular Health and Pelvic Inflammatory Disease Based on the 2013-2018 National Health and Nutrition Examination Survey Data
Shuangshuang XIE ; Yangxue YIN ; Liming YUAN ; Lingyun LIAO ; Qin XU ; Yanping ZHANG ; Rong ZHOU
Journal of Practical Obstetrics and Gynecology 2025;41(5):400-405
Objective:To analyze the relationship between Cardiovascular Health(CVH)and Pelvic Inflamma-tory Disease(PID)using data from the National Health and Nutrition Examination Surveys(NHANES).Meth-ods:Participants from the NHANES database were extracted based on the inclusion of PID,LE8 scores,and mul-tiple potential confounding factors,excluding those with missing variables.Multivariate Logistic regression analysis was conducted using R to assess the relationship between the CVH and the risk of PID.Additionally,subgroup a-nalysis and interaction tests were performed for age,race,poverty,marital status,education,BMI,menstrual regu-larity,and pregnancy history.Results:A total of 3934 female participants were included,with an average age of 39.95±11.42 years.Multivariable Logistic regression analysis showed that after adjusting for age,race,BMI,edu-cation level,marital status,household income,menstrual cycle,and pregnancy history,the risk of PID gradually decreased with an increase in CVH assessment metric LE8 scores(OR 0.98,95%CI 0.97-0.99,P<0.001).Stratified analysis indicated significant interactions between age,race,and marital status in the association be-tween CVH and PID(interaction P<0.05 for all),while household income level,education level,BMI,menstrual regularity,and pregnancy history showed no significant interactions with the CVH and PID association(P>0.05).Conclusions:There is a significant negative relationship between CVH and PID in adult women in the United states,suggesting that better cardiovascular health may help reduce the risk of PID.
8.Development, reliability, and validity of a treatment-related quality of life scale for Chinese patients with multiple myeloma
Chunyan SUN ; Zhen CAI ; Bing CHEN ; Lijuan CHEN ; Wenming CHEN ; Kaiyang DING ; Juan DU ; Rong FU ; Chengcheng FU ; Da GAO ; Guangxun GAO ; Yanjuan HE ; Jian HOU ; Ming JIANG ; Fei LI ; Jian LI ; Juan LI ; Zhenyu LI ; Aijun LIAO ; Jing LIU ; Jun LUO ; Jianmin LUO ; Yanping MA ; Jianqing MI ; Ting NIU ; Hongling PENG ; Yongping SONG ; Luqun WANG ; Rong ZHAN ; Xi ZHANG ; Yu HU
Chinese Journal of Hematology 2025;46(8):713-721
Objective:To develop a treatment-related quality of life scale for Chinese patients with multiple myeloma (MM) and to test its reliability and validity.Methods:The initial scale was constructed through a literature search, Delphi expert correspondence, and cognitive testing. This study conducted a preliminary survey of 379 patients with MM and a formal survey of 865 patients from the hematology departments of 155 hospitals nationwide from February 2024 to March 2024. The final scale was obtained after conducting item analysis and reliability and validity tests on the initial scale.Results:The constructed scale contains 36 items covering six domains: physiological, psychological, social, treatment side effects, general health, and others. In the preliminary survey, the Cronbach’s alpha coefficient of each item ranged from 0.597 to 0.939, and the test-retest reliability was 0.747 ( P<0.001). Exploratory factor analysis extracted eight common factors with a cumulative variance contribution of 60.058%. In the formal survey, the Cronbach’s alpha coefficient of each item ranged from 0.484 to 0.930, and the test-retest reliability was 0.835 ( P<0.001). Confirmatory factor analysis revealed a comparative fit index of 0.750, a root-mean-square error of approximation of 0.090, and a root-mean-square residual of 0.067. Conclusion:The treatment-related quality of life scale for Chinese patients with MM designed in this study exhibited good reliability and validity, reflecting the impact of treatment on the quality of life of patients. This scale can provide a reference to clinicians for assessing the disease status of patients.
9.Long-term effectiveness evaluation of the construction of “mosquito-free village” in Pujiang County
GUO Song ; HUANG Wenzhong ; SUN Jimin ; WU Hongzhao ; LIU Ying ; ZHANG Yanping ; REN Jiangping ; ZHANG Rong ; SHI Xuguang ; CHEN Enfu
Journal of Preventive Medicine 2024;36(5):374-377,382
Objective:
To evaluate the effectiveness for the construction of "mosquito-free village" in Xuejia Village, Pujiang County, Zhejiang Province, so as to provide the guidance for the construction of "mosquito-free village" in other rural areas.
Methods:
Density of adult mosquitoes in Xuejia Village was investigated using light trap method and density of larval mosquitoes was investigated using larval straw method from April to November each year. Totally 30 households of villagers were randomly selected, and their awareness rates of mosquito control knowledge, mosquito control behavior forming rates and satisfaction rates were surveyed through questionnaires. Investment during construction (from August 2016 to December 2018) and maintenance period (from 2019 to 2023) of "mosquito-free village" were investigated through data review and interviews. Long-term effects of "mosquito-free village" construction in Xuejia Village were evaluated in terms of mosquito density from 2016 to 2023, effectiveness of health education for villagers, satisfaction and investment.
Results:
Compared with the year 2016, the densities of adult and larval mosquitoes in Xuejia Village were significantly decreased from 2017 to 2023. The average monthly density of adult mosquitoes in 2023 decreased by 98.34%, and the average monthly 100 household index of larval mosquitoes decreased by 98.45% compared to 2016. The average monthly density of adult mosquitoes from 2019 to 2023 was less than or equal to one mosquito per light trap in a night, and the average monthly 100 household index of larval mosquitoes was less than or equal to five places per 100 households. The awareness rate of mosquito control knowledge was 93.33%, the behavior forming rate was 86.67%, and the satisfaction rate was 90.00%. By December 2023, the total investment during construction and maintenance period was 450 thousand Yuan, with an average annual investment of 60.7 thousand Yuan and average annual investment of 206.61 Yuan per household. The average annual investment during maintenance period was 36.2 thousand Yuan, and the average annual investment per household was 109.70 Yuan.
Conclusion
The mosquito density, effectiveness of health education for villagers and satisfaction of "mosquito-free village" in Xuejia Village all meet the evaluation criteria of "mosquito-free village", and the investment is reasonable, making it suitable for promotion to other rural areas.
10.Qingshen granules regulate mitophagy through miR-4516/SIAH3/PINK1 to attenuate renal fibrosis in rats
Kejun REN ; Hua JIN ; Dong WANG ; Yanping MAO ; Rong DAI ; Meng CHENG ; Yiping WANG ; Lei ZHANG
Chinese Journal of Pathophysiology 2024;40(8):1479-1487
AIM:To explore the mechanism by which Qingshen granules(QSG)intervene in the microRNA-4516(miR-4516)targeted regulation of the SIAH3/PINK1 axis,enhancing mitophagy and inhibiting renal fibrosis.METHODS:Male SD rats were randomly divided into three groups:control,model,and QSG groups.The QSG aqueous solution was administered via gavage once daily,4 mL each time,for 8 consecutive weeks.Blood creatinine levels were measured in each group.Hematoxylin-eosin(HE)and Masson staining were utilized to assess the degree of renal patholog-ical damage.Western blot analysis was performed to determine the expression levels of β-actin,PINK1,Parkin,SIAH3,VDAC1,Mfn1,Mfn2,OPA1,LC3B,and P62 proteins in renal tissue.RT-qPCR was used to detect the mRNA expres-sion level of SIAH3 in rat kidney tissue,and transmission electron microscopy was employed to observe mitochondrial dam-age in renal tissue.QSG-containing serum and transforming growth factor-β1(TGF-β1)were used to induce an HK-2 cell fibrosis model.The cells were divided into the following groups:normal cell(NC)group,model cell(MC)group,MC+miR-4516 mimics group,MC+miR-4516 NC+QSG group,MC+miR-4516 mimics+QSG group,and MC+QSG group.Cell activity in each group was detected using the CCK-8 method,and Western blot analysis was performed to determine E-cad-herin and α-SMA protein expression levels.The regulation of SIAH3 by miR-4516 was verified using a dual luciferase re-porter assay.RT-qPCR was used to detect the mRNA expression of miR-4516,SIAH3 mRNA,and PINK1.RESULTS:The results indicated that QSG intervention reduced fibrosis in rat renal tissue and HK-2 cells,decreased SIAH3 mRNA expression,increased PINK1 expression,and activated mitophagy in renal tissue.In vitro results confirmed that QSG can elevate miR-4516 expression,inhibit SIAH3 mRNA expression,promote PINK1 expression in HK-2 cells,and reduce the expression of the fibrosis marker protein α-SMA.CONCLUSION:In summary,this study preliminarily clarified the mechanism by which QSG intervention targets miR-4516 to regulate the SIAH3/PINK1 axis,thereby enhancing mitophagy and inhibiting renal fibrosis.


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