1.Protective Effect of Tongluo Baoshen Prescription-containing Serum on Lipopolysaccharide-induced Podocyte Injury of Rats
Yongfang LIU ; Tiantian YIN ; Huiyang LIU ; Rui HUANG ; Zhiying FENG ; Li ZHOU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(10):139-148
ObjectiveTo observe the effects of Tongluo Baoshen prescription (TLBS)-containing serum on the rat podocyte injury induced by lipopolysaccharide (LPS) and explore the potential mechanisms. MethodsSD rats were used to prepare the blank serum, losartan potassium-containing serum, and low-, medium-, and high-dose TLBS-containing sera. Rat podocytes were cultured in vitro, and the effects of drug-containing sera on podocyte viability were detected by the cell counting kit-8 (CKK-8) method. The optimal intervention volume fraction of drug-containing sera and the optimal concentration of LPS for inducing the podocyte injury were determined. Rat podocytes were grouped as follows: normal control (NC, 10% blank serum), model control (MC, 20.00 mg·L-1 LPS+10% black serum), losartan potassium (LP, 20.00 mg·L-1 LPS+10% losartan potassium-containing serum), low-dose TLBS (TLBS-L, 20.00 mg·L-1 LPS+10% low-dose TLBS-containing serum), medium-dose TLBS (TLBS-M, 20.00 mg·L-1 LPS+10% medium-dose TLBS-containing serum), and high-dose TLBS (TLBS-H, 20.00 mg·L-1 LPS+10% high-dose TLBS-containing serum), and the interventions lasted for 48 h. The ultrastructure of podocytes was observed under a transmission electron microscope. The podocyte apoptosis was detected by the terminal deoxynucleoitidyl transferase mediated nick-end labeling (TUNEL) kit. Immunofluorescence was used to detect the expression of gasdermin D N-terminal fragment (GSDMD-NT) in podocytes. The mRNA and protein levels of G protein-coupled receptor family C group 5 member B (GPRC5B), nuclear factor-κB (NF-κB) p50, NF-κB p52, NF-κB p65, Rel B, c-Rel, NOD-like receptor protein 3 (NLRP3), cysteinyl aspartate-specific protease-1 (Caspase-1), GSDMD-NT, interleukin (IL)-1β, IL-18, nephrin, integrin α3, and integrin β1 in podocytes were determined by real-time quaritiative polymerase chain reaction (Real-time PCR) and Western blot, respectively. ResultsCompared with the NC group, the MC group showed reduced podocyte protrusions and organelles, segmental missing of cell membranes, increased and swollen mitochondria, irregular nuclear membranes, light chromatin, increased TUNEL fluorescence-positive nuclei (P<0.01), obviously enhanced fluorescence intensity of GSDMD-NT, up-regulated mRNA and protein levels of GPRC5B, NF-κB p50, NF-κB p52, NF-κB p65, Rel B, c-Rel, NLRP3, caspase-1, GSDMD-NT, IL-1β, and IL-18 (P<0.01), and down-regulated mRNA and protein levels of nephrin, integrin α3, and integrin β1 (P<0.01) in podocytes. Compared with the MC group, the LP, TLBS-M, and TLBS-H groups showed improved ultrastructure of podocytes with increased protrusions, intact cell membranes, reduced organelles, and alleviated mitochondrial swelling, decreased TUNEL fluorescence-positive nuclei (P<0.01), weakened fluorescence intensity of GSDMD-NT, down-regulated mRNA and protein levels of GPRC5B, NF-κB p50, NF-κB p52, NF-κB p65, Rel B, c-Rel, NLRP3, caspase-1, GSDMD-NT, IL-1β, and IL-18 (P<0.01), and up-regulated mRNA and protein levels of nephrin, integrin α3, and integrin β1 (P<0.05, P<0.01). Moreover, the changes above were the most obvious in the TLBS-H group. ConclusionThe TLBS-containing serum can regulate the GPRC5B/NF-κB/NLRP3 pathway to inhibit pyroptosis, thereby ameliorating the podocyte injury induced by LPS.
2.Study on the Mechanism of Tongluo Baoshen Decoction Intervening IgA Nephropathy Rats
Yongfang LIU ; Rui HUANG ; Huiyang LIU ; Jianfeng DAI ; Yinghua LIU ; Xingyu LIU ; Zhiying FENG ; Tiantian YIN ; Li ZHOU
Chinese Journal of Information on Traditional Chinese Medicine 2025;32(8):105-113
Objective To observe the therapeutic effects of Tongluo Baoshen Prescription on IgA nephropathy rats;To explore its mechanism.Methods Totally 130 SD rats were randomly divided into a normal group(20 rats)and a modeling group(110 rats).After establishing the IgA nephropathy model,the modeling group was further randomly divided into the model group,losartan potassium group and Tongluo Baoshen Prescription low-,medium-and high-dosage groups,with 20 rats in each group.Tongluo Baoshen Prescription low-,medium-and high-dosage groups were administered at dosages of 5.3,10.6 and 21.2 g/kg respectively,the losartan potassium group was given with the dosage of 5 mg/kg,the normal group and model group were given an equivalent amount of distilled water by gavage once daily.Blood,urine and kidney tissue samples were collected after 4 and 8 weeks of administration.The hemoglobin content was detected,urinary IL-1β,integrin α3β1 and serum albumin contents were detected by ELISA,and the mRNA and protein expressions of NF-κB p52,NF-κB p65,Rel B,c-Rel and Desmin in renal tissue were detected by RT-qPCR and Western blot,the apoptosis of renal tissue at 8 weeks of administration was detected by TUNEL staining,the expression of C3c in renal tissue was detected by immunofluorescence,the co-expression of Gprc5b,Caspase-1,NLRP3 and Nephrin were detected by immunofluorescence double labeling,the expression of Gprc5b and Nephrin in renal tissue were detected by immunohistochemistry.Results Compared with the normal group,the model group rats showed significantly increased contents of urinary IL-1β and integrin α3β1(P<0.01),with further increases in urinary integrin α3β1 over time(P<0.05);the mRNA and protein expressions of NF-κB p52,NF-κB p65,Rel B,c-Rel and Desmin in renal tissue significantly increased(P<0.01),and the proportion of apoptosis-positive cells in renal tissue significantly increased(P<0.01),immunofluorescence results revealed that C3c exhibited granular and clumpy high-intensity deposition in the mesangial area and capillary loops of the glomeruli(P<0.01),immunofluorescence double labeling showed that the expression of Gprc5b,Caspase-1 and NLRP3 in renal tissue increased(P<0.01),while Nephrin expression significantly decreased(P<0.01),immunohistochemistry analysis showed that the positive expression of Gprc5b in renal tissue significantly increased(P<0.01),while Nephrin significantly decreased(P<0.01).Compared with the model group,the urinary IL-1β and integrin α3β1 contents in the losartan potassium group and Tongluo Baoshen Prescription groups reduced after 4 and 8 weeks of treatment(P<0.05,P<0.01);the mRNA and protein expressions of NF-κB p52,NF-κB p65,Rel B,c-Rel and Desmin in renal tissue decreased(P<0.05,P<0.01),and the proportion of apoptosis-positive cells in renal tissue decreased(P<0.05),the C3c deposition in renal tissue was diminished(P<0.05,P<0.01),immunofluorescence double labeling showed that the expression of Gprc5b,Caspase-1 and NLRP3 in renal tissue were decreased(P<0.05,P<0.01),while Nephrin expression increased(P<0.05,P<0.01),immunohistochemistry analysis showed that the positive expression of Gprc5b in renal tissue decreased(P<0.05,P<0.01),while Nephrin expression increased(P<0.05,P<0.01).Conclusion Tongluo Baoshen Prescription has a protective effect on the kidney of IgA nephropathy rats,and it can improve podocyte injury.The mechanism may be related to the inhibition of the Gprc5b/NF-κB/NLRP3 pathway.
3.Clinical analysis of a novel stent assisted ileal bypass for preventing anastomotic leakage in Da Vinci robot anus-preserving surgery of low rectal cancer
Jianing YAN ; Yongfang YIN ; Jiabin YANG ; Zhilong YAN ; Jianming XIE
Chinese Journal of General Surgery 2025;40(2):114-118
Objective:To explore the safety and effectiveness of ileal bypass with stent to prevent anastomotic leakage in Da Vinci robot anus-preserving surgery for low rectal cancer.Methods:From Dec 2021 to Sep 2024, the clinical data of 55 patients undergoing robot anus-preserving surgery and stent ileal bypass for low rectal cancer in the First Affiliated Hospital of Ningbo University was analyzed retrospectively.Results:All the 55 patients successfully completed the operation. Among them, 47 patients underwent low anterior resection and 8 patients did sphincter resection. The total operation time was (207±37)min, the operation time of ileal bypass was (24±3)min, and the intraoperative blood loss was (32±18)ml. After operation, the time of stent disintegration was (23.93± 2.87) days, and the time of catheter removal was (29.55 ± 3.21) days. By postoperative TNM staging, there were 12 cases in T1 stage, 26 cases in T2 and 17 cases in T3. The number of lymph node dissection was (16.84±4.50), 15 cases being positive for lymph node metastasis, 40 cases being negtive for lymph node metastasis, 36 cases were stage Ⅰ, 4 cases were stage Ⅱ and 15 cases were stage Ⅲ. The median follow-up time was 20 (2-35) months. Delayed anastomotic leakage occurred in 1 patient and anastomotic stenosis occurred in 2 patients.Conclusion:Stent ileal bypass is safe and effective to prevent anastomotic leakage in Da Vinci robot anus-preserving surgery for low rectal cancer.
4.A comparative study of asymmetrical prominent veins sign in stroke based on susceptibility maps and phase maps
Yongfang YIN ; Yan LIANG ; Yuhuan WANG ; Hao FANG ; Haibo LIU ; Yanling ZHAO
Journal of Practical Radiology 2025;41(10):1600-1603
Objective To explore the value of the minimum intensity projection(minIP)images generated by post-processing of susceptibility weighted imaging(SWI)and corrected phase image(CPI)in evaluating the asymmetrical prominent veins sign(APVS)in acute ischemic stroke.Methods A retrospective analysis was conducted on 86 patients with acute ischemic stroke.Group A underwent conventional SWI reconstruction to generate minIP images,while group B used CPI for re-reconstruction to produce minIP images.Both groups used the same scanning method but different post-processing techniques to generate two sets of images,with each group consisted of 86 patients.Two deputy chief physicians of imaging diagnostics scored subjectively with a double-blind 5-point method to compare the ability of the two groups to display APVS and analyze the display rate of APVS.Results The subjective scores of group B were significantly higher than those of group A,with a statistically significant difference(P<0.05).The display rates of APVS in groups A and B were 67.44%and 73.26%respectively.Group B had a higher display rate of APVS below the tentorium cerebelli than above it.Conclusion The minIP images generated by CPI post-processing can achieve the effects similar to phase difference enhanced imaging(PADRE),and is superior to SWI reconstruction method in displaying APVS.It can be used as a supplementary post-processing method when acute stroke shows poor APVS,which has practical clinical application value and can provide more imaging basis for clinical practice.
5.A comparative study of asymmetrical prominent veins sign in stroke based on susceptibility maps and phase maps
Yongfang YIN ; Yan LIANG ; Yuhuan WANG ; Hao FANG ; Haibo LIU ; Yanling ZHAO
Journal of Practical Radiology 2025;41(10):1600-1603
Objective To explore the value of the minimum intensity projection(minIP)images generated by post-processing of susceptibility weighted imaging(SWI)and corrected phase image(CPI)in evaluating the asymmetrical prominent veins sign(APVS)in acute ischemic stroke.Methods A retrospective analysis was conducted on 86 patients with acute ischemic stroke.Group A underwent conventional SWI reconstruction to generate minIP images,while group B used CPI for re-reconstruction to produce minIP images.Both groups used the same scanning method but different post-processing techniques to generate two sets of images,with each group consisted of 86 patients.Two deputy chief physicians of imaging diagnostics scored subjectively with a double-blind 5-point method to compare the ability of the two groups to display APVS and analyze the display rate of APVS.Results The subjective scores of group B were significantly higher than those of group A,with a statistically significant difference(P<0.05).The display rates of APVS in groups A and B were 67.44%and 73.26%respectively.Group B had a higher display rate of APVS below the tentorium cerebelli than above it.Conclusion The minIP images generated by CPI post-processing can achieve the effects similar to phase difference enhanced imaging(PADRE),and is superior to SWI reconstruction method in displaying APVS.It can be used as a supplementary post-processing method when acute stroke shows poor APVS,which has practical clinical application value and can provide more imaging basis for clinical practice.
6.Study on the Mechanism of Tongluo Baoshen Decoction Intervening IgA Nephropathy Rats
Yongfang LIU ; Rui HUANG ; Huiyang LIU ; Jianfeng DAI ; Yinghua LIU ; Xingyu LIU ; Zhiying FENG ; Tiantian YIN ; Li ZHOU
Chinese Journal of Information on Traditional Chinese Medicine 2025;32(8):105-113
Objective To observe the therapeutic effects of Tongluo Baoshen Prescription on IgA nephropathy rats;To explore its mechanism.Methods Totally 130 SD rats were randomly divided into a normal group(20 rats)and a modeling group(110 rats).After establishing the IgA nephropathy model,the modeling group was further randomly divided into the model group,losartan potassium group and Tongluo Baoshen Prescription low-,medium-and high-dosage groups,with 20 rats in each group.Tongluo Baoshen Prescription low-,medium-and high-dosage groups were administered at dosages of 5.3,10.6 and 21.2 g/kg respectively,the losartan potassium group was given with the dosage of 5 mg/kg,the normal group and model group were given an equivalent amount of distilled water by gavage once daily.Blood,urine and kidney tissue samples were collected after 4 and 8 weeks of administration.The hemoglobin content was detected,urinary IL-1β,integrin α3β1 and serum albumin contents were detected by ELISA,and the mRNA and protein expressions of NF-κB p52,NF-κB p65,Rel B,c-Rel and Desmin in renal tissue were detected by RT-qPCR and Western blot,the apoptosis of renal tissue at 8 weeks of administration was detected by TUNEL staining,the expression of C3c in renal tissue was detected by immunofluorescence,the co-expression of Gprc5b,Caspase-1,NLRP3 and Nephrin were detected by immunofluorescence double labeling,the expression of Gprc5b and Nephrin in renal tissue were detected by immunohistochemistry.Results Compared with the normal group,the model group rats showed significantly increased contents of urinary IL-1β and integrin α3β1(P<0.01),with further increases in urinary integrin α3β1 over time(P<0.05);the mRNA and protein expressions of NF-κB p52,NF-κB p65,Rel B,c-Rel and Desmin in renal tissue significantly increased(P<0.01),and the proportion of apoptosis-positive cells in renal tissue significantly increased(P<0.01),immunofluorescence results revealed that C3c exhibited granular and clumpy high-intensity deposition in the mesangial area and capillary loops of the glomeruli(P<0.01),immunofluorescence double labeling showed that the expression of Gprc5b,Caspase-1 and NLRP3 in renal tissue increased(P<0.01),while Nephrin expression significantly decreased(P<0.01),immunohistochemistry analysis showed that the positive expression of Gprc5b in renal tissue significantly increased(P<0.01),while Nephrin significantly decreased(P<0.01).Compared with the model group,the urinary IL-1β and integrin α3β1 contents in the losartan potassium group and Tongluo Baoshen Prescription groups reduced after 4 and 8 weeks of treatment(P<0.05,P<0.01);the mRNA and protein expressions of NF-κB p52,NF-κB p65,Rel B,c-Rel and Desmin in renal tissue decreased(P<0.05,P<0.01),and the proportion of apoptosis-positive cells in renal tissue decreased(P<0.05),the C3c deposition in renal tissue was diminished(P<0.05,P<0.01),immunofluorescence double labeling showed that the expression of Gprc5b,Caspase-1 and NLRP3 in renal tissue were decreased(P<0.05,P<0.01),while Nephrin expression increased(P<0.05,P<0.01),immunohistochemistry analysis showed that the positive expression of Gprc5b in renal tissue decreased(P<0.05,P<0.01),while Nephrin expression increased(P<0.05,P<0.01).Conclusion Tongluo Baoshen Prescription has a protective effect on the kidney of IgA nephropathy rats,and it can improve podocyte injury.The mechanism may be related to the inhibition of the Gprc5b/NF-κB/NLRP3 pathway.
7.Clinical analysis of a novel stent assisted ileal bypass for preventing anastomotic leakage in Da Vinci robot anus-preserving surgery of low rectal cancer
Jianing YAN ; Yongfang YIN ; Jiabin YANG ; Zhilong YAN ; Jianming XIE
Chinese Journal of General Surgery 2025;40(2):114-118
Objective:To explore the safety and effectiveness of ileal bypass with stent to prevent anastomotic leakage in Da Vinci robot anus-preserving surgery for low rectal cancer.Methods:From Dec 2021 to Sep 2024, the clinical data of 55 patients undergoing robot anus-preserving surgery and stent ileal bypass for low rectal cancer in the First Affiliated Hospital of Ningbo University was analyzed retrospectively.Results:All the 55 patients successfully completed the operation. Among them, 47 patients underwent low anterior resection and 8 patients did sphincter resection. The total operation time was (207±37)min, the operation time of ileal bypass was (24±3)min, and the intraoperative blood loss was (32±18)ml. After operation, the time of stent disintegration was (23.93± 2.87) days, and the time of catheter removal was (29.55 ± 3.21) days. By postoperative TNM staging, there were 12 cases in T1 stage, 26 cases in T2 and 17 cases in T3. The number of lymph node dissection was (16.84±4.50), 15 cases being positive for lymph node metastasis, 40 cases being negtive for lymph node metastasis, 36 cases were stage Ⅰ, 4 cases were stage Ⅱ and 15 cases were stage Ⅲ. The median follow-up time was 20 (2-35) months. Delayed anastomotic leakage occurred in 1 patient and anastomotic stenosis occurred in 2 patients.Conclusion:Stent ileal bypass is safe and effective to prevent anastomotic leakage in Da Vinci robot anus-preserving surgery for low rectal cancer.
8.Role of NLRP3 Inflammasome in IgA Nephropathy and Chinese Medicine Intervention: A Review
Yongfang LIU ; Li ZHOU ; Huiyang LIU ; Rui HUANG ; Zhiying FENG ; Tiantian YIN
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(6):269-279
IgA nephropathy is recognized as the most common primary glomerular disease, with up to 20%-40% of patients developing end-stage kidney disease within 20 years of onset. The deposition of IgA1-containing immune complexes targeting glycosylation defects in the mesangial region and the subsequent inflammation caused by T lymphocyte activation are considered as the main causes of IgA nephropathy, and innate immunity is also involved in the pathogenesis. Nucleotide-binding oligomerization domain (NOD)-like receptor protein 3 (NLRP3) is a newly discovered pattern recognition receptor expressed in renal intrinsic cells such as renal tubular epithelial cells, mesangial cells, and podocytes. Activated by external stimuli, NLRP3 can form NLRP3 inflammasomes with apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC). The NLRP3 inflammasome can activate cysteine aspartate-specific protease-1 (Caspase-1), causing the maturation and release of interleukin-18 (IL-18) and interleukin-1β (IL-1β) involved in inflammation. Increasing evidence has suggested that NLRP3 inflammasomes are involved in the pathogenesis and progression of IgA nephropathy and associated with the damage of renal intrinsic cells such as podocytes, mesangial cells, endothelial cells, and renal tubular epithelial cells. Chinese medicine can regulate inflammatory cytokines and their signaling pathways by acting on NLRP3 inflammasomes and related molecules, exerting therapeutic effects on IgA nephropathy. This article introduces the role of NLRP3 inflammasomes in IgA nephropathy and reviews the clinical and experimental research progress of Chinese medicine intervention in IgA nephropathy via NLRP3 inflammasomes, aiming to provide a reference for further research and application of Chinese medicine intervention in the NLRP3 inflammasome as a new therapeutic target.
9.Da Vinci robot assisted total gastrectomy plus hand-sewn esophagojejunostomy for gastric carcinaoma
Wanbo HUANG ; Jianming XIE ; Jiabin YANG ; Yongfang YIN ; Haixiang DING ; Xiuchong YU ; Zhilong YAN
Chinese Journal of General Surgery 2024;39(10):758-763
Objective:To evaluate the safety and feasibility of robot assisted total gastrectomy plus hand-sewn esophagojejunostomy.Methods:The clinical data of 72 patients diagnosed with gastric cancer and undergoing total gastrectomy at the First Affiliated Hospital of Ningbo University from Nov 2021 to May 2024 were retrospectively analyzed. They were divided into two groups: robot-assisted total gastrectomy (RATG) group, consisting of 30 patients, and laparoscopic assisted total gastrectomy (LATG) group, consisting of 42 patients . In the RATG group, the digestive tract was reconstructed by manual suture under the robot scope and Roux-Y reconstruction was performed . In LATG group, digestive tract reconstruction was performed using an in vitro stapler and Roux-Y. The clinicopathological data, perioperative indexes, and postoperative follow-up data of both groups were observed and analyzed.Results:All 72 patients successfully completed the operation without conversion to open laparotomy. The total operation time in RATG group was longer than that in LATG group [(235.2±25.8) min vs. (200.7±40.6) min, t=4.099, P<0.05)].RATG was superior to LATG group in terms of digestive tract reconstruction time, postoperative fluid intake time and hospitalization days,the difference was statistically significant [(36.9±3.0) min vs.(39.4±4.5) min, (4.2±0.5) d vs. (5.2±0.6) d、(9.5±1.6) d vs. (10.8±2.4)d, t=-2.554,-7.135,-2.595, all P<0.05]; In terms of postoperative pathology, the number of lymph node dissection in RATG group was higher than that in LATG group [(29.8±6.2) vs. (26.3±7.5), t=2.197, P<0.05]. Conclusion:The application of delayed disconnection hand-sewn esophagojejunostomy in Da Vinci robot total gastrectomy is safe and feasible for cure-intent total gastrectomy in patients of gastric carcinoma.
10.Prediction of peritoneal free cancer cells in gastric cancer patients by golden-angle radial sampling dynamic contrast-enhanced magnetic resonance imaging
YIN XUEQING ; RUAN XINZHONG ; ZHU YONGMENG ; YIN YONGFANG ; HUANG RUI ; LIANG CHAO
Journal of Zhejiang University. Science. B 2024;25(7):617-627
Objective:Peritoneal free cancer cells can negatively impact disease progression and patient outcomes in gastric cancer.This study aimed to investigate the feasibility of using golden-angle radial sampling dynamic contrast-enhanced magnetic resonance imaging(GRASP DCE-MRI)to predict the presence of peritoneal free cancer cells in gastric cancer patients.Methods:All enrolled patients were consecutively divided into analysis and validation groups.Preoperative magnetic resonance imaging(MRI)scans and perfusion were performed in patients with gastric cancer undergoing surgery,and peritoneal lavage specimens were collected for examination.Based on the peritoneal lavage cytology(PLC)results,patients were divided into negative and positive lavage fluid groups.The data collected included clinical and MR information.A nomogram prediction model was constructed to predict the positive rate of peritoneal lavage fluid,and the validity of the model was verified based on data from the verification group.Results:There was no statistical difference between the proportion of PLC-positive cases predicted by GRASP DCE-MR and the actual PLC test.MR tumor stage,tumor thickness,and perfusion parameter Tofts-Ketty model volume transfer constant(Ktrans)were independent predictors of positive peritoneal lavage fluid.The nomogram model featured a concordance index(C-index)of 0.785 and 0.742 for the modeling and validation groups,respectively.Conclusions:GRASP DCE-MR could effectively predict peritoneal free cancer cells in gastric cancer patients.The nomogram model constructed using these predictors may help clinicians to better predict the risk of peritoneal free cancer cells being present in gastric cancer patients.

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