1.Mechanism of Qiju Dihuang Wan in Treatment of Diabetic Retinopathy Based on NLRP3/Caspase-1/GSDMD Signaling Pathway
Ge ZHANG ; Chunlin ZHAO ; Li PAN ; Ying ZHANG ; Zehai CHEN ; Siya ZHANG ; Yiming FANG ; Wei SHI
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(20):11-21
ObjectiveTo investigate the protective effect of Qiju Dihuang Wan on diabetic retinopathy (DR) and its regulatory mechanism on the NOD-like receptor protein 3/Cysteinyl aspartate-specific proteinase-1/Gasdermin D (NLRP3/Caspase-1/GSDMD) signaling pathway. MethodsHuman retinal microvascular endothelial cells (HRMEC) were divided into the blank group, model group, and Qiju Dihuang Wan low- and high-dose groups. Except for the blank group, all groups were induced with 40 mmol·L-1 high glucose, and the treatment groups were supplemented with 6% and 12% Qiju Dihuang Wan-containing serum, respectively, based on the induction. After 24 h of culture, the levels of interleukin (IL)-1β and IL-18 were measured by enzyme-linked immunosorbent assay (ELISA), pyroptosis-related protein expression was detected by immunofluorescence and Western blot, and the mRNA levels of NLRP3/Caspase-1/GSDMD signaling pathway-related molecules were determined by real-time polymerase chain reaction (Real-time PCR). The mice were randomly divided into the blank group, model group, Qiju Dihuang Wan low-, medium-, and high-dose groups (11, 22, 44 g·kg-1·d-1), and calcium dobesilate group (0.13 g·kg-1·d-1). Except for the blank group, all groups were intraperitoneally injected with streptozotocin to establish the DR model. After successful modeling, the treatment groups received corresponding doses of Qiju Dihuang Wan crude drug or calcium dobesilate suspension by gavage once daily, while the blank group and model group received an equal volume of normal saline, for 8 consecutive weeks. Fundus changes were examined by optical coherence tomography, optical coherence tomography angiography, and fundus fluorescein angiography. Retinal pathological morphology was observed by hematoxylin-eosin staining. Real-time PCR was used to detect the mRNA levels of NLRP3, Caspase‑1, GSDMD, IL‑1β, and IL‑18 in retinal tissue, and Western blot was used to detect the protein expression of NLRP3, ASC, cleaved Caspase‑1, and GSDMD‑N. ResultsCell experiments showed that compared with the blank group, the model group showed significantly decreased HRMEC viability (P<0.01), and significantly increased levels of IL-1β, IL-18, as well as increased protein expression of NLRP3, cleaved Caspase-1, and GSDMD-N (P<0.01). Compared with the model group, the above indicators were significantly reduced in the high-dose Qiju Dihuang Wan groups (P<0.05, P<0.01). Animal experiments showed that compared with the blank group, the model group had elevated blood glucose (P<0.01), decreased retinal thickness and vascular density (P<0.01), up‑regulated mRNA expression of NLRP3, Caspase‑1, GSDMD, IL‑1β and IL‑18 in retinal tissue (P<0.01), and up‑regulated protein expression of NLRP3, ASC, cleaved Caspase‑1 and GSDMD‑N (P<0.01). Compared with the model group, the Qiju Dihuang Wan medium- and high‑dose groups showed decreased blood glucose (P<0.01), increased retinal thickness and vascular density (P<0.01), alleviated retinal pathological damage, and down‑regulated mRNA expression of NLRP3, Caspase‑1, GSDMD, IL‑1β, IL‑18, as well as protein expression of NLRP3, ASC, cleaved Caspase‑1, and GSDMD‑N (P<0.05, P<0.01). ConclusionQiju Dihuang Wan may exert protective effects against diabetic retinopathy by inhibiting the activation of the NLRP3/Caspase-1/GSDMD signaling pathway and alleviating high glucose-induced retinal pyroptosis.
2.Mechanism of Qiju Dihuang Wan in Treatment of Diabetic Retinopathy Based on NLRP3/Caspase-1/GSDMD Signaling Pathway
Ge ZHANG ; Chunlin ZHAO ; Li PAN ; Ying ZHANG ; Zehai CHEN ; Siya ZHANG ; Yiming FANG ; Wei SHI
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(20):11-21
ObjectiveTo investigate the protective effect of Qiju Dihuang Wan on diabetic retinopathy (DR) and its regulatory mechanism on the NOD-like receptor protein 3/Cysteinyl aspartate-specific proteinase-1/Gasdermin D (NLRP3/Caspase-1/GSDMD) signaling pathway. MethodsHuman retinal microvascular endothelial cells (HRMEC) were divided into the blank group, model group, and Qiju Dihuang Wan low- and high-dose groups. Except for the blank group, all groups were induced with 40 mmol·L-1 high glucose, and the treatment groups were supplemented with 6% and 12% Qiju Dihuang Wan-containing serum, respectively, based on the induction. After 24 h of culture, the levels of interleukin (IL)-1β and IL-18 were measured by enzyme-linked immunosorbent assay (ELISA), pyroptosis-related protein expression was detected by immunofluorescence and Western blot, and the mRNA levels of NLRP3/Caspase-1/GSDMD signaling pathway-related molecules were determined by real-time polymerase chain reaction (Real-time PCR). The mice were randomly divided into the blank group, model group, Qiju Dihuang Wan low-, medium-, and high-dose groups (11, 22, 44 g·kg-1·d-1), and calcium dobesilate group (0.13 g·kg-1·d-1). Except for the blank group, all groups were intraperitoneally injected with streptozotocin to establish the DR model. After successful modeling, the treatment groups received corresponding doses of Qiju Dihuang Wan crude drug or calcium dobesilate suspension by gavage once daily, while the blank group and model group received an equal volume of normal saline, for 8 consecutive weeks. Fundus changes were examined by optical coherence tomography, optical coherence tomography angiography, and fundus fluorescein angiography. Retinal pathological morphology was observed by hematoxylin-eosin staining. Real-time PCR was used to detect the mRNA levels of NLRP3, Caspase‑1, GSDMD, IL‑1β, and IL‑18 in retinal tissue, and Western blot was used to detect the protein expression of NLRP3, ASC, cleaved Caspase‑1, and GSDMD‑N. ResultsCell experiments showed that compared with the blank group, the model group showed significantly decreased HRMEC viability (P<0.01), and significantly increased levels of IL-1β, IL-18, as well as increased protein expression of NLRP3, cleaved Caspase-1, and GSDMD-N (P<0.01). Compared with the model group, the above indicators were significantly reduced in the high-dose Qiju Dihuang Wan groups (P<0.05, P<0.01). Animal experiments showed that compared with the blank group, the model group had elevated blood glucose (P<0.01), decreased retinal thickness and vascular density (P<0.01), up‑regulated mRNA expression of NLRP3, Caspase‑1, GSDMD, IL‑1β and IL‑18 in retinal tissue (P<0.01), and up‑regulated protein expression of NLRP3, ASC, cleaved Caspase‑1 and GSDMD‑N (P<0.01). Compared with the model group, the Qiju Dihuang Wan medium- and high‑dose groups showed decreased blood glucose (P<0.01), increased retinal thickness and vascular density (P<0.01), alleviated retinal pathological damage, and down‑regulated mRNA expression of NLRP3, Caspase‑1, GSDMD, IL‑1β, IL‑18, as well as protein expression of NLRP3, ASC, cleaved Caspase‑1, and GSDMD‑N (P<0.05, P<0.01). ConclusionQiju Dihuang Wan may exert protective effects against diabetic retinopathy by inhibiting the activation of the NLRP3/Caspase-1/GSDMD signaling pathway and alleviating high glucose-induced retinal pyroptosis.
3.Amlodipine promotes autophagy by inhibiting
Wei Zhao ; Deping Xu ; Kainan Liao ; Chunlin Cai ; Dandan Zang ; Haisheng Zhou
Acta Universitatis Medicinalis Anhui 2025;60(7):1179-1186
Objective :
To explore the effects of the antihypertensive drug Amlodipine on calcium influx and autoph- agy in human podocytes ( HPC) .
Methods :
HPC cells were routinely cultured in vitro. HPC cells were treated with angiotensin Ⅱ ( Ang Ⅱ ) ,the L-type Ca2 + blocker Amlodipine alone or in combination.The Ca2 + imaging system was used to detect the transient changes in the intracellular Ca2 + flux of HPC cells in real time after drug treatment.Western blot was employed to detect the changes in the ratio of autophagy marker proteins LC3B-Ⅱ/ LC3B-Ⅰ , and the expression levels of Beclin-1,P62,as well as apoptosis-related proteins Bcl-2 and Bax.Flow cytometry was used to detect the number of Fluo-4AM positive cells at 488 nm to analyze the level of intracellular Ca2 + influx in HPC cells.Lyso-Tracker Green live cell staining was applied to analyze the fluorescence intensity of lysosomes.Flow cytometry was also used to detect the apoptosis rate of HPC cells.
Results :
Compared with the control group,in the Ang Ⅱ group,the transient Ca2 + flux and the number of Fluo-4AM positive cells increased significantly (P<0. 001) .The ratio of autophagy marker proteins LC3B-Ⅱ/ LC3B-Ⅰ (P<0. 001) and the pro- tein expression of Beclin-1 (P<0. 01) decreased significantly,while the expression of P62 increased (P<0. 01) . The fluorescence intensity of lysosomes weakened (P<0. 05) ,the apoptosis rate increased (P<0. 0001) ,the ex- pression of apoptosis-related protein Bcl-2 decreased (P <0. 01 ) ,and the protein level of Bax increased (P < 0. 001) .Compared with the control group,in the Amlodipine group,the number of Fluo-4AM positive cells de- creased significantly (P<0. 001) ,the ratio of LC3B-Ⅱ/ LC3B-Ⅰ (P<0. 001) and the protein expression of Bec- lin-1 (P<0. 001) increased,the protein expression of P62 decreased (P<0. 05) ,the fluorescence intensity of ly- sosomes enhanced (P<0. 01) ,the apoptosis rate decreased (P<0. 01) ,the protein expression of Bcl-2 increased (P<0. 001) ,and the protein level of Bax decreased (P<0. 001) .Compared with the Ang Ⅱ group,in the Ang Ⅱ + Amlodipine group,the number of Fluo-4AM positive cells decreased significantly (P<0. 001) ,the ratio of LC3B-Ⅱ/ LC3B-Ⅰ (P<0. 01) and the protein expression of Beclin-1 increased (P<0. 05) ,the protein level of P62 decreased (P<0. 01) ,the fluorescence intensity of lysosomes increased (P <0. 05) ,the apoptosis rate de- creased (P<0. 001) ,the protein expression of Bcl-2 increased (P <0. 001 ) ,and the protein level of Bax de- creased (P<0. 001) .
Conclusion
Amlodipine inhibits calcium influx,promotes autophagy and inhibits apoptosis in human podocytes,which is useful in preventing the development of hypertensive nephropathy.
4.Screening of soil biocontrol bacteria and evaluation of their control effects on Fusarium head blight of wheat.
Dongfang WANG ; Xinxin ZHAI ; Chunlin YANG ; Huilan ZHANG ; Jie WU ; Zerong SONG ; Pan ZHAO ; Yu CHI
Chinese Journal of Biotechnology 2025;41(10):3764-3773
Fusarium head blight (FHB), caused by Fusarium graminearum, not only leads to severe yield losses but also poses a threat to food safety due to the mycotoxins produced by the pathogen. Since this disease is preventable but not curable, the current control mainly relies on chemical fungicides, the long-term use of which may lead to pathogen resistance and environmental pollution. To develop green control methods, we screened 13 biocontrol strains from the rhizosphere soil of wheat, among which strain No. 12 (identified as Pythium aphanidermatum) showed significant antifungal effects. In the plate confrontation test, this strain reduced the colony diameter of the pathogen by 69.2% (1.47 mm vs. 4.78 mm in the control group), with an inhibition rate of 77% (P < 0.01). Microscopic observation revealed obvious deformations in the pathogen hyphae, suggesting a lysing effect. The coleoptile experiment further confirmed that the pre-treatment with this strain reduced the incidence rate to 0. These findings provide new candidate strains for the biocontrol of FHB and offer a scientific basis for reducing the use of chemical fungicides and promoting sustainable agricultural development.
Triticum/growth & development*
;
Fusarium/growth & development*
;
Plant Diseases/prevention & control*
;
Soil Microbiology
;
Pest Control, Biological/methods*
;
Pythium/physiology*
;
Biological Control Agents
;
Rhizosphere
;
Fungicides, Industrial
5.Expert Consensus on Classification of Hand Degloving Injures and Emergency Repair of Avulsion Skin
Jihui JU ; Gang ZHAO ; Yongjun RUI ; Xin WANG ; Weiyang GAO ; Xiaoheng DING ; Qingtang ZHU ; Xianyou ZHENG ; Yongqing XU ; Shanlin CHEN ; Juyu TANG ; Lei XU ; Jianxi HOU ; Huaqiao WANG ; Jingyi MI ; Haifeng SHI ; Shusen CUI ; Chunlin HOU ; Liqiang GU
Chinese Journal of Microsurgery 2025;48(2):121-134
Hand degloving injury represents one of the most severe forms of hand trauma, characterised by challenging treatment and a complex prognostic outcome. It is crucial to effectively utilise the degloved tissues in emergency or primary repair of a hand degloving injury. This consensus provides a comprehensive review of the existing literature on definition, classification, emergency assessment, debridement, judgment of skin viability, in situ repair of the degloved skin, and adjunctive treatment for degloving injury of hand. Based on conclusion of both domestic and international experiences, this expert consensus on the classification of hand degloving injury and the emergency repair with the avulsed skin is established, aiming to provide a guidance to surgeons on standardised treatment strategy and improve the management of hand degloving injury.
6.Construction of a new staging system for stage N3 gastric cancer based on the metastatic lymph node ratio
Hongyu ZHANG ; Guanghui LIU ; Yanwei YE ; Chunlin ZHAO ; Yang FU
Chinese Journal of General Surgery 2025;40(2):123-130
Objective:To explore the cut-off value of metastatic lymph node ratio (LNR) for stage N3 gastric cancer and construct a new TNM staging system to predict prognosis.Methods:Clinical data of 4 291 patients from Jan 2004 to Dec 2020 in the SEER database and 567 patients from Jan 2016 to Dec 2020 in the First Affiliated Hospital of Zhengzhou University with stage N3 gastric cancer were collected. A new TNrM staging system and a nomogram model were constructed based on the optimal LNR cut-off value and compared with the 8th TNM staging model in terms of prognostic discrimination, prognostic prediction accuracy, and clinical usefulness.Results:The optimal cut-off value of LNR was 0.5. A TNrM staging system was constructed by combining the Nr stage with the T stage. Univariate and multivariate COX regression analyses showed that the TNrM staging system was a significant prognostic factor (all P<0.05). Based on the 8th TNM and TNrM staging system, two nomograms were constructed in the training set and externally validated in the validation set. Compared with the TNM staging model, the TNrM staging model had a larger C-index and area of time-dependent ROC curve(AUC) (training set: 3-year AUC: 66.5 vs. 74.4, 5-year AUC: 68.9 vs. 75.3; validation set: 3-year AUC: 62.3 vs. 73.1, 5-year AUC: 62.6 vs. 75.8), its overall survival prediction curves were closer to the ideal curve in the calibration curve, and its clinical net benefit was greater in the decision curves. Conclusions:Stage N3 gastric cancer patients with a metastatic lymph node ratio >0.5 have a poor prognosis. The TNrM staging nomogram model constructed based on the lymph node ratio has better prognostic discrimination ability and prediction accuracy and more clinical net benefits compared to the 8th edition of TNM staging nomogram model.
7.Latent class of unmet need trajectories during chemotherapy for gastric cancer patients and their impact on economic toxicity of chemotherapy
Ruiqin LIU ; Shulan GAO ; Huanan WANG ; Chunlin ZHAO
Chinese Journal of Modern Nursing 2025;31(17):2270-2274
Objective:To explore the level of unmet need during chemotherapy for gastric cancer patients, analyze its dynamic trajectory and impact on the economic toxicity of chemotherapy.Methods:Convenience sampling was used to select 385 gastric cancer patients who received chemotherapy from June 2022 to December 2023 at the First Affiliated Hospital of Zhengzhou University. The level of unmet need for patients' first chemotherapy cycle (C 1) , second chemotherapy cycle (C 2) and third chemotherapy cycle (C 3) and economic toxicity at the end of chemotherapy were investigated separately. The latent class growth model (LCGM) was used to analyze classes of unmet need trajectories and to compare differences in patient economic toxicity across classes. Results:Finally 368 patients completed all data collection, and the effective response rate of survey respondents was 95.58%. As chemotherapy progressed, patients' unmet need scores gradually increased, and the unmet need scores of C 1, C 2, and C 3 were (35.54±5.44) , (47.32±6.45) , and (58.19±11.81) , respectively, which were categorized into three latent classes of medium-low level rapid increase group, medium level gradual increase group, and medium level stable group. There were 122 cases (33.15%) in medium-low level rapid increase group, 167 cases (45.38%) in medium level gradual increase group, and 79 cases (21.47%) in medium level stable group. The lowest economic toxicity score was found in medium-low level rapid increase group, followed by medium level gradual increase group, and the highest in medium level stable group, and the difference in economic toxicity scores among the three groups was statistically significant ( P<0.05) . The percentage of patients with positive economic toxicity in the three groups was 45.90% (56/122) , 28.14% (47/167) , and 18.99% (15/79) , respectively, with a statistically significant difference ( P<0.05) . Conclusions:Unmet needs of patients during chemotherapy for gastric cancer show a moderate to high level and gradually increase with the course of chemotherapy. The trajectory can be divided into three latent classes of medium-low level rapid increase group, medium level gradual increase group, and medium level stable group. There are differences in economic toxicity for patients with different classes of unmet need trajectories, with patients in increase or rapid increase groups facing severe economic toxicity.
8.Latent class of unmet need trajectories during chemotherapy for gastric cancer patients and their impact on economic toxicity of chemotherapy
Ruiqin LIU ; Shulan GAO ; Huanan WANG ; Chunlin ZHAO
Chinese Journal of Modern Nursing 2025;31(17):2270-2274
Objective:To explore the level of unmet need during chemotherapy for gastric cancer patients, analyze its dynamic trajectory and impact on the economic toxicity of chemotherapy.Methods:Convenience sampling was used to select 385 gastric cancer patients who received chemotherapy from June 2022 to December 2023 at the First Affiliated Hospital of Zhengzhou University. The level of unmet need for patients' first chemotherapy cycle (C 1) , second chemotherapy cycle (C 2) and third chemotherapy cycle (C 3) and economic toxicity at the end of chemotherapy were investigated separately. The latent class growth model (LCGM) was used to analyze classes of unmet need trajectories and to compare differences in patient economic toxicity across classes. Results:Finally 368 patients completed all data collection, and the effective response rate of survey respondents was 95.58%. As chemotherapy progressed, patients' unmet need scores gradually increased, and the unmet need scores of C 1, C 2, and C 3 were (35.54±5.44) , (47.32±6.45) , and (58.19±11.81) , respectively, which were categorized into three latent classes of medium-low level rapid increase group, medium level gradual increase group, and medium level stable group. There were 122 cases (33.15%) in medium-low level rapid increase group, 167 cases (45.38%) in medium level gradual increase group, and 79 cases (21.47%) in medium level stable group. The lowest economic toxicity score was found in medium-low level rapid increase group, followed by medium level gradual increase group, and the highest in medium level stable group, and the difference in economic toxicity scores among the three groups was statistically significant ( P<0.05) . The percentage of patients with positive economic toxicity in the three groups was 45.90% (56/122) , 28.14% (47/167) , and 18.99% (15/79) , respectively, with a statistically significant difference ( P<0.05) . Conclusions:Unmet needs of patients during chemotherapy for gastric cancer show a moderate to high level and gradually increase with the course of chemotherapy. The trajectory can be divided into three latent classes of medium-low level rapid increase group, medium level gradual increase group, and medium level stable group. There are differences in economic toxicity for patients with different classes of unmet need trajectories, with patients in increase or rapid increase groups facing severe economic toxicity.
9.Expert Consensus on Classification of Hand Degloving Injures and Emergency Repair of Avulsion Skin
Jihui JU ; Gang ZHAO ; Yongjun RUI ; Xin WANG ; Weiyang GAO ; Xiaoheng DING ; Qingtang ZHU ; Xianyou ZHENG ; Yongqing XU ; Shanlin CHEN ; Juyu TANG ; Lei XU ; Jianxi HOU ; Huaqiao WANG ; Jingyi MI ; Haifeng SHI ; Shusen CUI ; Chunlin HOU ; Liqiang GU
Chinese Journal of Microsurgery 2025;48(2):121-134
Hand degloving injury represents one of the most severe forms of hand trauma, characterised by challenging treatment and a complex prognostic outcome. It is crucial to effectively utilise the degloved tissues in emergency or primary repair of a hand degloving injury. This consensus provides a comprehensive review of the existing literature on definition, classification, emergency assessment, debridement, judgment of skin viability, in situ repair of the degloved skin, and adjunctive treatment for degloving injury of hand. Based on conclusion of both domestic and international experiences, this expert consensus on the classification of hand degloving injury and the emergency repair with the avulsed skin is established, aiming to provide a guidance to surgeons on standardised treatment strategy and improve the management of hand degloving injury.
10.Construction of a new staging system for stage N3 gastric cancer based on the metastatic lymph node ratio
Hongyu ZHANG ; Guanghui LIU ; Yanwei YE ; Chunlin ZHAO ; Yang FU
Chinese Journal of General Surgery 2025;40(2):123-130
Objective:To explore the cut-off value of metastatic lymph node ratio (LNR) for stage N3 gastric cancer and construct a new TNM staging system to predict prognosis.Methods:Clinical data of 4 291 patients from Jan 2004 to Dec 2020 in the SEER database and 567 patients from Jan 2016 to Dec 2020 in the First Affiliated Hospital of Zhengzhou University with stage N3 gastric cancer were collected. A new TNrM staging system and a nomogram model were constructed based on the optimal LNR cut-off value and compared with the 8th TNM staging model in terms of prognostic discrimination, prognostic prediction accuracy, and clinical usefulness.Results:The optimal cut-off value of LNR was 0.5. A TNrM staging system was constructed by combining the Nr stage with the T stage. Univariate and multivariate COX regression analyses showed that the TNrM staging system was a significant prognostic factor (all P<0.05). Based on the 8th TNM and TNrM staging system, two nomograms were constructed in the training set and externally validated in the validation set. Compared with the TNM staging model, the TNrM staging model had a larger C-index and area of time-dependent ROC curve(AUC) (training set: 3-year AUC: 66.5 vs. 74.4, 5-year AUC: 68.9 vs. 75.3; validation set: 3-year AUC: 62.3 vs. 73.1, 5-year AUC: 62.6 vs. 75.8), its overall survival prediction curves were closer to the ideal curve in the calibration curve, and its clinical net benefit was greater in the decision curves. Conclusions:Stage N3 gastric cancer patients with a metastatic lymph node ratio >0.5 have a poor prognosis. The TNrM staging nomogram model constructed based on the lymph node ratio has better prognostic discrimination ability and prediction accuracy and more clinical net benefits compared to the 8th edition of TNM staging nomogram model.


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