1.Mechanisms of Jiangtang No. 3 Prescription in Alleviating Adipose Tissue Insulin Resistance in Diabetic Rats via TLR4/NF-κB/NLRP3 Signaling Pathway-mediated Inflammation
Tongxun WANG ; Lantian LIU ; Runqi LI ; Haoxiang LI ; Yi ZHAO ; Tian TIAN ; Rufeng MA ; Sihua GAO ; Dandan ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(4):109-117
ObjectiveTo observe the effects of Jiangtang No. 3 prescription on inflammatory pathways and insulin resistance-related indicators in rats with type 2 diabetes mellitus (T2DM), and to elucidate its molecular mechanism in combating diabetes. MethodsA T2DM rat model was established using a high-fat diet combined with intraperitoneal injection of streptozotocin (STZ). Successfully modeled rats were randomly assigned to the model group, metformin group, and low-, medium-, and high-dose Jiangtang No. 3 prescription groups, and a normal group was also set. Daily gavage was administered for 8 weeks as follows: metformin at 0.1 g·kg-1·d-1, Jiangtang No. 3 prescription granules at 1.62, 3.24, 6.48 g·kg-1·d-1 for the respective dose groups, and sterile water for the normal and model groups. Rat body weight, fasting blood glucose (FBG), oral glucose tolerance test (OGTT), and insulin tolerance test (ITT) were measured. After drug intervention, enzyme-linked immunosorbent assay (ELISA) was used to determine serum levels of total cholesterol (TC), triglycerides (TG), low-density lipoprotein cholesterol (LDL), high-density lipoprotein cholesterol (HDL), non-esterified fatty acids (NEFA), interleukin (IL)-1β, IL-18, and insulin (INS). Hematoxylin-eosin (HE) staining was used to observe morphological changes in adipose tissue. Real-time quantitative PCR was used to detect the mRNA expression of Toll-like receptor 4 (TLR4), nuclear factor-κB (NF-κB), NOD-like receptor protein 3 (NLRP3), Caspase-1, IL-1β, IL-18, and gasdermin D (GSDMD) in adipose tissue. Western blot was used to measure the corresponding protein expression levels. ResultsCompared with the model group, Jiangtang No. 3 prescription groups exhibited significantly increased body weight (P<0.05, P<0.01), significantly reduced FBG (P<0.05, P<0.01), significant reductions in TC, TG, NEFA, and LDL (P<0.05, P<0.01), and a significant increase in HDL (P<0.01). Serum levels of inflammatory mediators IL-1β and IL-18 were significantly decreased (P<0.01), the homeostatic model assessment of insulin resistance (HOMA-IR) index was significantly reduced (P<0.05, P<0.01), and adipose tissue pathology was improved. The protein expression levels of TLR4, NF-κB, NLRP3, Caspase-1, IL-1β, IL-18, and GSDMD were markedly decreased (P<0.05, P<0.01), and the mRNA expression levels of these indicators were also significantly downregulated (P<0.05, P<0.01). Some effects were superior to those of the positive control drug metformin, and certain indicators exhibited dose-dependent improvements. ConclusionT2DM rats display significant inflammatory responses, disordered glucose and lipid metabolism, and insulin resistance. Jiangtang No. 3 prescription effectively suppresses inflammatory mediators, improves glucose and lipid metabolism and insulin resistance, and ameliorates pathological changes in adipose tissue. Its mechanism may be related to the regulation of the TLR4/NF-κB/NLRP3 signaling pathway in visceral adipose tissue, thereby influencing downstream inflammatory mediators.
2.Analysis of the correlation between splenic changes and hepatic encephalopathy after transjugular intrahepatic portosystemic shunt
Jun’an LIN ; Jingqin MA ; Rufeng CHEN ; Zhiping YAN
Chinese Journal of Clinical Medicine 2026;33(1):45-51
Objective To explore the correlation between changes in spleen volume and stiffness before and after transjugular intrahepatic portosystemic shunt (TIPS) and the occurrence of hepatic encephalopathy. Methods A retrospective study was conducted on 104 patients who underwent TIPS in the Department of Interventional Therapy, Zhongshan Hospital, Fudan University, from January 1, 2023 to July 30, 2024. Spleen volume and stiffness were measured before and after surgery. Patients were divided into the overt hepatic encephalopathy (OHE) group and the non-OHE group. Univariate and multivariate logistic regression analyses were used to identify factors correlated with OHE after TIPS. The predictive values of the related factors of OHE were evaluated using receiver operating characteristic (ROC) curves. Results OHE occurred in 34 patients (32.69%). Compared with the non-OHE group, the OHE group had a smaller percentage reduction in spleen volume after TIPS (P<0.001), but there was no statistically significant difference in the percentage reduction in spleen stiffness between the two groups. Multivariate logistic regression analysis revealed that the percentage reduction in spleen volume was correlated with postoperative OHE (OR=0.90, P<0.001). ROC analysis showed that the area under the curve (AUC) for spleen volume reduction percentage in predicting postoperative OHE was 0.773, superior to spontaneous portosystemic shunt (SPSS; Z=2.088, P=0.037). Conclusions Reduction in spleen volume after TIPS may be a protective factor against hepatic encephalopathy, while changes in spleen stiffness were not found to be correlated with its occurrence.
3.The effect of reducing platelet reactive oxygen species on decreasing of the refrigerated platelets being phagocyted
Rufeng Xie ; Zhicheng Wang ; Jie Sun ; Yiming Yang ; Jie Yang
Acta Universitatis Medicinalis Anhui 2025;60(1):123-129
Objective :
To explore the effect of the addition of N-acetylcysteine ( NAC) on the abnormal increase of reactive oxygen species ( ROS) during platelet cold storage,and to clarify its function of preventing the platelets from being recognized and cleared by macrophages and hepatocytes.
Methods :
Platelets Concentrates were divided into room temperature group ( 22C) ,cold storage group ( 4C) and NAC addition group.In NAC addition group, the platelets were supplemented with 5 mmol / L ( N5) and 25 mmol / L ( N25) NAC.After 5-day storage,the levels of ROS,activation markers and other index of platelets in each group were detected by flow cytometry. Platelet phagocytosis was detected by PMA-activated THP-1 cells or by primary cultured HepG2 cells.
Results :
After 5 days of storage,ROS increased significantly in 4C group than those in 22C group ( P<0. 05) ,and after NAC addi- tion,ROS level reduced significantly ( P<0. 05) .The expression of CD62P and PS and the exposure of β-GlcNAc on platelets in N5 group significantly decreased compared with those in 4C group ( P<0. 05) .The platelet phago- cytosis by THP-1 and HepG2 cells were also significantly lower in N5 group than those in 4C group ( P<0. 05) . However,the addition of 5 mmol / L NAC did not significantly affect the count,pH,CD42b expression,β-Gal ex- posure and coagulation function of cold-stored platelet after 5-day storage.
Conclusion
The addition of NAC to re- frigerated platelets can significantly decrease the level of platelet ROS and effectively reduce their phagocytosis by cells,suggesting that NAC addition may protect refrigerated platelets from being cleared by phagocytosis after trans- fusion.
4.Stability of 5-FU in whole blood and a clinical sampling and delivery procedures for TDM
Yongqing WEN ; Wenjuan WANG ; Yu BAI ; Rufeng LIU ; Xu MA
China Pharmacy 2025;36(23):2963-2968
OBJECTIVE To investigate the stability of 5-fluorouracil (5-FU) in human blood and to establish a standardized clinical sampling and delivery procedure for therapeutic drug monitoring (TDM) of 5-FU. METHODS The EDTA-anticoagulated whole blood was used as the matrix to prepare stability assessment samples of 5-FU at both low (200 ng/mL) and high (5 000 ng/mL) concentrations (with groups without stabilizer and with 1% volume ratio of stabilizer). The stability assessment samples were placed under room temperature ([ 25±2) ℃] and refrigerated (2-8 ℃) conditions, with sampling at 0, 0.5, 1, 2, 4, 7, and 24 h. After vortexing and centrifugation, the upper plasma layer was collected; proteins were precipitated using methanol, and the concentration of 5-FU in plasma was determined by liquid chromatography-tandem mass spectrometry. Based on the whole blood stability results, clinical sampling and delivery procedures were established. RESULTS The concentration of 5-FU in blank whole blood samples without stabilizers was significantly lower than that in samples with stabilizers (P<0.05). However, varying volumes (10, 25, 50 μL) of stabilizers had no significant effect on the measured concentrations of 5-FU in stability assessment samples with low and high concentrations (P>0.05). Without the addition of a stabilizer, low- and high-concentration 5-FU whole blood samples remained stable at room temperature for 0.5 h and 1 h, respectively, and under refrigeration for 2 h and 7 h, respectively. After the addition of a 1% stabilizer, the whole blood samples remained stable for up to 24 h under both room temperature and refrigerated conditions. Based on these findings, the following procedure was established: after collection, whole blood samples could be temporarily stored at room temperature (≤0.5 h) or at 4 ℃ (≤2 h), and transported at 2-8 ℃. Upon delivery to the laboratory, a 1% volume ratio of stabilizer must be added immediately, followed by centrifugation within 24 h. The resulting plasma should be stored at -20 ℃ . CONCLUSIONS 5-FU in whole blood exhibits poor stability at room temperature. Refrigeration at 2-8 ℃ slightly improves stability , but degradation still occurs rapidly. Adding a stabilizer at a 1% volume ratio significantly prolongs the refrigerated storage time. The established sampling and transport procedure for 5-FU TDM innovatively introduces the stabilizer addition step at the laboratory sample reception stage (rather than immediately after blood draw). This approach ensures analytical quality while offering greater adaptability to real-world clinical sampling conditions, significantly improving practical feasibility.
5.Short-term efficacy of rituximab in children with calcineurin inhibitor resistant steroid resistant nephrotic syndrome
Sicheng YU ; Jialu LIU ; Jiaojiao LIU ; Xiaoyan FANG ; Jing CHEN ; Qianfan MIAO ; Xiaoshan TANG ; Zhiqing ZHANG ; Chunyan WANG ; Rufeng DAI ; Xinli HAN ; Yihui ZHAI ; Hong XU ; Qian SHEN
Chinese Journal of Pediatrics 2025;63(2):185-189
Objective:To investigate the short-term efficacy and safety of rituximab (RTX) in children with calcineurin inhibitor (CNI) resistant steroid resistant nephrotic syndrome (SRNS).Methods:A retrospective case analysis was conducted. Thirteen children with CNI resistant SRNS who were regularly treated with RTX (375 mg/m 2 per dose (maximum dose 500 mg), 1 dose per week, a total of 4 doses) in Department of Nephrology, Children′s Hospital of Fudan University from January 2016 to December 2023 were enrolled. The general data, disease related information, urinary protein/creatinine, serum albumin, blood creatinine before RTX treatment, immunosuppressants, adverse events, and monthly urinary protein/creatinine, serum albumin, and blood creatinine indexes within 6 months after RTX treatment were collected. The changes of urinary protein/creatinine, serum albumin and estimated glomerular filtration rate (eGFR) before and after RTX at 3 and 6 months were analyzed by using paired sample t test and Wilcoxon signed-rank test. Results:Among the 13 patients, 8 were male and 5 were female. The age of disease onset was 4.0 (2.9, 6.8) years and the age of RTX treatment was 9.8 (5.9, 13.6) years. There were 8 cases of focal segmental glomerulosclerosis, 3 cases of minimal change disease and 2 cases of mesangial proliferative glomerulonephritis. No clinically significant gene variation was detected in 12 cases and the other one did not receive gene test. Before RTX treatment, 11 cases were in chronic kidney disease stage G1, and 1 case each was in stage G2 and stage G3. Ten children completed 4 doses of RTX treatment, 1 patient completed 3 doses, and 2 patients completed 2 doses. Urinary protein/creatinine in 13 children at 3 and 6 months after RTX treatment was significantly lower than baseline (0.60 (0.13, 2.04), 0.49 (0.28, 1.10) vs. 1.44 (0.76, 4.11) mg/mg, Z=-2.34, -2.34, both P<0.05), and serum albumin was significantly higher than baseline ((35±8), (34±7) vs. (30±6) g/L, t=2.30, 2.60, both P<0.05). The eGFR at 6 months after RTX treatment was not significantly different from the baseline ((110±32) vs. (113±35) ml/(min·1.73 m 2), t=-0.76, P>0.05)). No serious adverse reactions occurred in this study. Conclusion:RTX could reduce urinary protein and increase serum albumin in short-term treatment in children with CNI resistant SRNS without significant side effects.
6.Short-term efficacy of rituximab in children with calcineurin inhibitor resistant steroid resistant nephrotic syndrome
Sicheng YU ; Jialu LIU ; Jiaojiao LIU ; Xiaoyan FANG ; Jing CHEN ; Qianfan MIAO ; Xiaoshan TANG ; Zhiqing ZHANG ; Chunyan WANG ; Rufeng DAI ; Xinli HAN ; Yihui ZHAI ; Hong XU ; Qian SHEN
Chinese Journal of Pediatrics 2025;63(2):185-189
Objective:To investigate the short-term efficacy and safety of rituximab (RTX) in children with calcineurin inhibitor (CNI) resistant steroid resistant nephrotic syndrome (SRNS).Methods:A retrospective case analysis was conducted. Thirteen children with CNI resistant SRNS who were regularly treated with RTX (375 mg/m 2 per dose (maximum dose 500 mg), 1 dose per week, a total of 4 doses) in Department of Nephrology, Children′s Hospital of Fudan University from January 2016 to December 2023 were enrolled. The general data, disease related information, urinary protein/creatinine, serum albumin, blood creatinine before RTX treatment, immunosuppressants, adverse events, and monthly urinary protein/creatinine, serum albumin, and blood creatinine indexes within 6 months after RTX treatment were collected. The changes of urinary protein/creatinine, serum albumin and estimated glomerular filtration rate (eGFR) before and after RTX at 3 and 6 months were analyzed by using paired sample t test and Wilcoxon signed-rank test. Results:Among the 13 patients, 8 were male and 5 were female. The age of disease onset was 4.0 (2.9, 6.8) years and the age of RTX treatment was 9.8 (5.9, 13.6) years. There were 8 cases of focal segmental glomerulosclerosis, 3 cases of minimal change disease and 2 cases of mesangial proliferative glomerulonephritis. No clinically significant gene variation was detected in 12 cases and the other one did not receive gene test. Before RTX treatment, 11 cases were in chronic kidney disease stage G1, and 1 case each was in stage G2 and stage G3. Ten children completed 4 doses of RTX treatment, 1 patient completed 3 doses, and 2 patients completed 2 doses. Urinary protein/creatinine in 13 children at 3 and 6 months after RTX treatment was significantly lower than baseline (0.60 (0.13, 2.04), 0.49 (0.28, 1.10) vs. 1.44 (0.76, 4.11) mg/mg, Z=-2.34, -2.34, both P<0.05), and serum albumin was significantly higher than baseline ((35±8), (34±7) vs. (30±6) g/L, t=2.30, 2.60, both P<0.05). The eGFR at 6 months after RTX treatment was not significantly different from the baseline ((110±32) vs. (113±35) ml/(min·1.73 m 2), t=-0.76, P>0.05)). No serious adverse reactions occurred in this study. Conclusion:RTX could reduce urinary protein and increase serum albumin in short-term treatment in children with CNI resistant SRNS without significant side effects.
7.Analysis on the homogeneity of clinical basic skills teaching based on OSCE exam scores
Jia XU ; Guoli WANG ; Rufeng ZHANG ; Jing WANG ; Geng WANG ; Yu YANG
Journal of Shenyang Medical College 2024;26(2):217-220
Objective:To identify and improve the weakness in different clinical practice teaching hospital,and enhance the quality of practical teaching.Methods:A total of 291 trainees majoring in clinical medicine in grade 2016 in a medical college in Shenyang who practiced in different clinical teaching hospitals and participated in the objective structured clinical examination(OSCE)were enrolled.The OSCE scores was analyzed with one way ANOVA and Kruskal-Wallis statistic methods to identify the weakness in clinical practice teaching and improve the teaching quality.Results:In the standardized patient consultation and physical examination results,the passing rate and average score of H hospital was the lowest.The average score of trainees in H hospital was statistically significant compared to that in A,B,F,G,K,and L hospitals(P<0.01).In terms of skill operation scores,H hospital had the lowest pass rate and average score.The average score of interns in H hospital was statistically significant compared to that in A,D,E,F,G,I,J,K hospitals(P<0.01).In the interpretation of auxiliary examination results,the passing rate and average score of H hospital was the lowest.The average score of interns in A and H hospital was significantly different from that of B and J hospitals(P<0.01).Conclusions:There are great differences in the practice effect of students in different clinical teaching hospitals.Medical colleges and universities should strengthen the management of clinical teaching hospitals to ensure the homogeneity of clinical practice teaching quality.
8.Clinical efficacy on the treatment of thoracolumbar fracture using a homemade bone graft funnel for injured vertebral bone graft combined with percutaneous pedicle screw internal fixation
Long JIA ; Xiangqian LI ; Rufeng GAO
Chinese Journal of Spine and Spinal Cord 2024;34(5):476-483
Objectives:To investigate the clinical efficacy of using a homemade minimally invasive percuta-neous bone graft funnel for the treatment of thoracolumbar vertebral fractures with injured vertebral bone graft in combination with percutaneous pedicle screw intemal fixation.Methods:Clinical data from 26 patients with thoracolumbar spine fractures without nerve injury admitted and treated between January 2020 and June 2021 were retrospectively analyzed.There were 13 males and 13 females,aged 51.23±9.56 years(33-65 years old).All the patients were single-segment injury,and there were 2 cases of T12,8 cases of L1,8 cases of L2,5 cases of L3,and 3 cases of L4.Internal fixation was performed using percutaneous pedicle screw fixa-tion across the injured vertebrae,followed by intemal implantation of autologous iliac bone into the injured vertebrae via a homemade minimally invasive bone graft funnel.The operative time,intraoperative bleeding and complications were recorded;The visual analogue scale(VAS)of pain,Oswestry disability index(ODI),com-pression ratios of the anterior and middle heights of the injured vertebrae,and sagittal Cobb angle were ob-served and statistically analyzed before operation and at 1 week after operation and final follow-up.Results:All the patients were successfully operated with a follow-up of 12-22 months(15.19±2.51 months).The opera-tive time was 84.62±12.88min,blood loss was 55.58±12.44mL.There were no complications such as wound infection and spinal cord injury,and one patient presented with caudal adventitial loosening of a screwed pedicle.Compared with the preoperative period,the postoperative 1 week and final follow-up anterior height compression rate[(6.89±7.25)%,(10.28±7.50)%]and mid-height compression rate[(7.11±5.75)%,(10.63±6.24)%]of the injured vertebrae,Cobb angle(4.38°±7.77°,2.14°±7.78°),VAS score(3.35±0.56,1.73±0.45)and ODI[(41.96±3.82)%,(13.77±2.42)%]were significantly improved(P<0.05).Compared with 1 week postoperatively,at the final follow-up,the patients had a slight increase in the compression rate of the anterior[(10.28±7.50)%]and middle[(10.63±6.24)%]heights of the injured vertebral body,a slight decrease in the Cobb angle(2.14°±7.78°)and a significant improvement in VAS scores(1.73±0.45)and ODI[(13.77±2.42)%](P<0.05).Conclusions:For patients with thoracolumbar fractures without nerve injury,using a homemade minimally invasive bone graft funnel for transforaminal bone grafting in combination with percutaneous pedicle screws can effectively restore the vertebral body height and correct the kyphosis deformity.
9.Comparison of liver and adrenal transplantation models of neuroblastoma
Hongxia CHEN ; Zhigang TAN ; Huiran LIN ; Luping FENG ; Chuya ZHENG ; Wenfeng LIAO ; Rufeng ZENG ; Jinxin LIU ; Zhenjian ZHUO
Chinese Journal of Comparative Medicine 2024;34(4):100-108
Objective A neuroblastoma(NB)liver transplantation model was established and compared with the adrenal orthotopic transplantation model to explore its characteristics.Methods 5× 105 SK-N-SH cells were implanted along the long axis of the left lobe of mouse livers with a micro-injection needle.The growth,metastasis,expression of related genes,and histopathological changes of tumors were detected after the modeling.Results The tumor formation rate in mice inoculated with tumor cells reached 100%after 21 days,and tumor growth,metastasis,related gene expression changes,and pathological characteristics were apparent.Conclusions In this study,a neuroblastoma liver transplantation model was successfully constructed via a relatively simple surgical method to provide a more suitable choice for future scientific NB experiments.
10.The molecular mechanisms of Jiang Tang San Hao Formula alleviating inflammatory responses in diabetic mice via the NLPR3 inflammasome
Yi ZHAO ; Runqi LI ; Bingrui XU ; Zimengwei YE ; Fangfang MO ; Tian TIAN ; Guangtong DONG ; Rufeng MA ; Xiaoda YANG ; Sihua GAO ; Dandan ZHAO
Journal of Beijing University of Traditional Chinese Medicine 2024;47(11):1541-1549
Objective This study aimed to observe the effect of Jiang Tang San Hao Formula(JTSHF)on systemic and intestinal inflammation,as well as on the NLRP3 inflammasome in type 2 diabetic mice(T2DM),and to elucidate its anti-diabetic molecular mechanisms.Methods Four-week-old male C57BL/6 N mice were used to establish the T2DM model using a high-fat diet combined with streptozotocin injection.The diabetic mice were randomly divided into the model,metformin,and JTSHF groups.A control group was also set to provide baseline comparisons.Each group of mice was orally administered with the corresponding medication daily.The metformin group was orally administered with 0.20 g/kg metformin,the JTSHF group was orally administered with 4.26 g/kg JTSHF,and the control group and model group were orally administered with an equal amount of sterile water continuously for 8 weeks.After an 8-week drug intervention via gavage,the lipopolysaccharide(LPS),tumor necrosis factor-alpha(TNF-α),interleukin 1 beta(IL-1β),and interleukin 6(IL-6)serum and colon levels were quantified using an enzyme-linked immunosorbent assay(ELISA).The pathological morphology of the colon was observed using hematoxylin and eosin staining.NOD-like receptor protein 3(NLRP3),apoptosis-associated speck-like protein containing a caspase recruitment domain(ASC),caspase-1,zonula occludens-1(ZO-1),occludin,and G-protein coupled receptor 43(GPR43)protein expression in the colon were assessed using immunohistochemistry.The mRNA expression levels of NLRP3,ASC,caspase-1,ZO-1,Occludin,and GPR43 in the colon were detected using Real-time PCR.Results The ELISA data revealed significant differences in inflammatory markers among the groups.Compared with the model group,the JTSHF group exhibited notably reduced LPS,TNF-α,IL-1β,and IL-6 levels(P<0.05).Moreover,compared with the model group,JTSHF treatment upregulated ZO-1,occludin,and GPR43 protein and mRNA expression in the colon and downregulated NLRP3,ASC,and Caspase-1 protein and mRNA expression(P<0.05).Conclusion The inflammatory reaction of T2DM mice is apparent.JTSHF effectively alleviates the systemic and intestinal inflammatory response of T2DM mice by inhibiting the NLRP3 inflammasome and repairing the intestinal mucosal barrier,highlighting the potential molecular mechanisms of the anti-diabetes effects of JTSHF.


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