1.Shaoyaotang Containing Serum Mediates Fas/FasL Pathway to Inhibit Lipopolysaccharide Induced Inflammation and Apoptosis of Caco-2 Cells
Yuting YANG ; Dongsheng WU ; Hui CAO ; Yu ZHANG ; Nianjia XIE ; Bo ZOU ; Daguang CHEN ; Erle LIU ; Yi LU ; Zhaowen LYU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(13):62-69
ObjectiveTo investigate the effects of different concentrations of Shaoyaotang-containing serum on lipopolysaccharide (LPS)-induced inflammation of human colorectal adenocarcinoma (Caco-2) cells by inhibiting apoptosis via activating the tumor necrosis factor (TNF) receptor superfamily member 6 (Fas)/Fas ligand (FasL) pathway. MethodsCaco-2 cells were allocated into blank, model (LPS, 10 mg·L-1), Shaoyaotang-containing serum (5%, 10%, 15%, 20%), and Fas inhibitor (KR-33493, 20 mmol·L-1) groups. Except the blank group, the other groups were stimulated with 10 mg·L-1 LPS for 24 h for the modeling of inflammation. After successful modeling, the blank, Fas inhibitor, and model groups were treated with blank serum, and the Shaoyaotang-containing serum groups were treated with the serum samples at corresponding concentrations for 24 h. The Fas inhibitor group was subjected to KR-33493 pretreatment for 1 h. Cell proliferation and viability were examined by the cell-counting kit-8 (CCK-8) method. The levels of interleukin (IL)-6, IL-1β, and TNF-α were measured by enzyme-linked immunosorbent assay. Apoptosis was detected by flow cytometry. The protein and mRNA levels of Fas, FasL, cysteinyl aspartate-specific proteinase (Caspase)-3, Caspase-9, B-cell lymphoma 2 (Bcl-2), and Bcl-2-associated X protein (Bax) were determined by Western blot and Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR), respectively. ResultsCompared with the blank group, the model group presented a decrease in cell survival rate (P<0.01). Compared with that in the model group, the cell survival rate showed no significant change in the 5% Shaoyaotang-containing serum group but increased in the 10%, 15%, and 20% Shaoyaotang-containing serum groups (P<0.01). Since there was no statistical difference between the 5% Shaoyaotang-containing serum group and the model group, 10%, 15%, and 20% Shaoyaotang-containing sera were selected for the follow-up study. Compared with the blank group, the model group showed risen levels of IL-6, IL-1β, and TNF-α (P<0.01), an increased apoptosis rate (P<0.01), up-regulated protein and mRNA levels of Fas, FasL, Caspase-3, Caspase-9, and Bax (P<0.01), and down-regulated protein and mRNA levels of Bcl-2 (P<0.01). Compared with the model group, the Fas inhibitor group and the 10%, 15%, and 20% Shaoyaotang-containing serum groups showed declined levels of IL-6, IL-1β, and TNF-α (P<0.01), decreased apoptosis rates (P<0.01), down-regulated protein and mRNA levels of Fas, FasL, Caspase-3, Caspase-9, and Bax (P<0.05, P<0.01), and up-regulated protein and mRNA levels of Bcl-2 (P<0.05, P<0.01). In addition, the 15% and 20% Shaoyaotang-containing serum groups had lower levels of IL-6, IL-1β, and TNF-α (P<0.05, P<0.01), lower apoptosis rates (P<0.05, P<0.01), lower protein and mRNA levels of Fas, FasL, Caspase-3, Caspase-9, and Bax (P<0.05, P<0.01), and higher protein and mRNA levels of Bcl-2 (P<0.05, P<0.01) than the 10% Shaoyaotang-containing serum group. ConclusionThe Shaoyaotang-containing serum can reduce the content of inflammatory factors in Caco-2 cells, down-regulate the protein and mRNA levels of Fas, FasL, Caspase-3, Caspase-9, and Bax, and up-regulate the protein and mRNA levels of Bcl-2 under the intervention of LPS by regulating the Fas/FasL pathway and inhibiting the apoptosis of intestinal epithelial cells in ulcerative colitis.
2.Shaoyaotang Containing Serum Mediates Fas/FasL Pathway to Inhibit Lipopolysaccharide Induced Inflammation and Apoptosis of Caco-2 Cells
Yuting YANG ; Dongsheng WU ; Hui CAO ; Yu ZHANG ; Nianjia XIE ; Bo ZOU ; Daguang CHEN ; Erle LIU ; Yi LU ; Zhaowen LYU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(13):62-69
ObjectiveTo investigate the effects of different concentrations of Shaoyaotang-containing serum on lipopolysaccharide (LPS)-induced inflammation of human colorectal adenocarcinoma (Caco-2) cells by inhibiting apoptosis via activating the tumor necrosis factor (TNF) receptor superfamily member 6 (Fas)/Fas ligand (FasL) pathway. MethodsCaco-2 cells were allocated into blank, model (LPS, 10 mg·L-1), Shaoyaotang-containing serum (5%, 10%, 15%, 20%), and Fas inhibitor (KR-33493, 20 mmol·L-1) groups. Except the blank group, the other groups were stimulated with 10 mg·L-1 LPS for 24 h for the modeling of inflammation. After successful modeling, the blank, Fas inhibitor, and model groups were treated with blank serum, and the Shaoyaotang-containing serum groups were treated with the serum samples at corresponding concentrations for 24 h. The Fas inhibitor group was subjected to KR-33493 pretreatment for 1 h. Cell proliferation and viability were examined by the cell-counting kit-8 (CCK-8) method. The levels of interleukin (IL)-6, IL-1β, and TNF-α were measured by enzyme-linked immunosorbent assay. Apoptosis was detected by flow cytometry. The protein and mRNA levels of Fas, FasL, cysteinyl aspartate-specific proteinase (Caspase)-3, Caspase-9, B-cell lymphoma 2 (Bcl-2), and Bcl-2-associated X protein (Bax) were determined by Western blot and Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR), respectively. ResultsCompared with the blank group, the model group presented a decrease in cell survival rate (P<0.01). Compared with that in the model group, the cell survival rate showed no significant change in the 5% Shaoyaotang-containing serum group but increased in the 10%, 15%, and 20% Shaoyaotang-containing serum groups (P<0.01). Since there was no statistical difference between the 5% Shaoyaotang-containing serum group and the model group, 10%, 15%, and 20% Shaoyaotang-containing sera were selected for the follow-up study. Compared with the blank group, the model group showed risen levels of IL-6, IL-1β, and TNF-α (P<0.01), an increased apoptosis rate (P<0.01), up-regulated protein and mRNA levels of Fas, FasL, Caspase-3, Caspase-9, and Bax (P<0.01), and down-regulated protein and mRNA levels of Bcl-2 (P<0.01). Compared with the model group, the Fas inhibitor group and the 10%, 15%, and 20% Shaoyaotang-containing serum groups showed declined levels of IL-6, IL-1β, and TNF-α (P<0.01), decreased apoptosis rates (P<0.01), down-regulated protein and mRNA levels of Fas, FasL, Caspase-3, Caspase-9, and Bax (P<0.05, P<0.01), and up-regulated protein and mRNA levels of Bcl-2 (P<0.05, P<0.01). In addition, the 15% and 20% Shaoyaotang-containing serum groups had lower levels of IL-6, IL-1β, and TNF-α (P<0.05, P<0.01), lower apoptosis rates (P<0.05, P<0.01), lower protein and mRNA levels of Fas, FasL, Caspase-3, Caspase-9, and Bax (P<0.05, P<0.01), and higher protein and mRNA levels of Bcl-2 (P<0.05, P<0.01) than the 10% Shaoyaotang-containing serum group. ConclusionThe Shaoyaotang-containing serum can reduce the content of inflammatory factors in Caco-2 cells, down-regulate the protein and mRNA levels of Fas, FasL, Caspase-3, Caspase-9, and Bax, and up-regulate the protein and mRNA levels of Bcl-2 under the intervention of LPS by regulating the Fas/FasL pathway and inhibiting the apoptosis of intestinal epithelial cells in ulcerative colitis.
3.Empirical and modified hemostatic resuscitation for liver blast injury combined with seawater immersion: A preliminary study.
Haoyang YANG ; Chenglin DAI ; Dongzhaoyang ZHANG ; Can CHEN ; Zhao YE ; Xin ZHONG ; Yijun JIA ; Renqing JIANG ; Wenqiong DU ; Zhaowen ZONG
Chinese Journal of Traumatology 2025;28(3):220-225
PURPOSE:
To compare the effects of empirical and modified hemostatic resuscitation for liver blast injury combined with seawater immersion.
METHODS:
Thirty rabbits were subjected to liver blast injury combined with seawater immersion, and were then divided into 3 groups randomly (n = 10 each): group A (no treatment after immersion), group B (empirical resuscitation with 20 mL hydroxyethyl starch, 50 mg tranexamic acid, 25 IU prothrombin complex concentrate and 50 mg/kg body weight fibrinogen concentrate), and group C (modified resuscitation with additional 10 IU prothrombin complex concentrate and 20 mg/kg body weight fibrinogen concentrate based on group B). Blood samples were gathered at specified moments for assessment of thromboelastography, routine coagulation test, and biochemistry. Mean arterial pressure, heart rate, and survival rate were also documented at each time point. The Kolmogorov-Smirnov test was used to examine the normality of data distribution. Multigroup comparisons were conducted with one-way ANOVA.
RESULTS:
Liver blast injury combined with seawater immersion resulted in severe coagulo-fibrinolytic derangement as indicated by prolonged prothrombin time (s) (11.53 ± 0.98 vs. 7.61 ± 0.28, p<0.001), activated partial thromboplastin time (APTT) (s) (33.48 ± 6.66 vs. 18.23 ± 0.89, p<0.001), reaction time (R) (min) (5.85 ± 0.96 vs. 2.47 ± 0.53, p<0.001), decreased maximum amplitude (MA) (mm) (53.20 ± 5.99 vs. 74.92 ± 5.76, p<0.001) and fibrinogen concentration (g/L) (1.19 ± 0.29 vs. 1.89 ± 0.32, p = 0.003), and increased D-dimer concentration (mg/L) (0.38 ± 0.32 vs. 0.05 ± 0.03, p = 0.005). Both empirical and modified hemostatic resuscitation could improve the coagulo-fibrinolytic states and organ function, as indicated by shortened APTT and R values, decreased D-dimer concentration, increased fibrinogen concentration and MA values, lower concentration of blood urea nitrogen and creatine kinase-MB in group B and group C rabbits in comparison to that observed in group A. Further analysis found that the R values (min) (4.67 ± 0.84 vs. 3.66 ± 0.98, p = 0.038), APTT (s) (23.16 ± 2.75 vs. 18.94 ± 1.05, p = 0.001), MA (mm) (60.10 ± 4.74 vs. 70.21 ± 3.01, p < 0.001), and fibrinogen concentration (g/L) (1.68 ± 0.21 vs. 1.94 ± 0.16, p = 0.013) were remarkably improved in group C than in group B at 2 h and 4 h after injury. In addition, the concentration of blood urea nitrogen (mmol/L) (24.11 ± 1.96 vs. 21.00 ± 3.78, p = 0.047) and creatine kinase-MB (U/L) (85.50 ± 13.60 vs. 69.74 ± 8.56, p = 0.013) were lower in group C than in group B at 6 h after injury. The survival rates in group B and group C were significantly higher than those in group A at 4 h and 6 h after injury (p < 0.001), however, there were no statistical differences in survival rates between group B and group C at each time point.
CONCLUSIONS
Modified hemostatic resuscitation could improve the coagulation parameters and organ function better than empirical hemostatic resuscitation.
Animals
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Rabbits
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Resuscitation/methods*
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Liver/injuries*
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Seawater
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Blast Injuries/therapy*
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Fibrinogen/administration & dosage*
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Male
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Tranexamic Acid/administration & dosage*
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Immersion
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Hydroxyethyl Starch Derivatives/administration & dosage*
4.Resuscitation effect of novel lyophilized platelets combined with lactated Ringer's solution in rabbits with hemorrhagic shock and seawater immersion
Chenglin DAI ; Wenqiong DU ; Ting TANG ; Haoyang YANG ; Liyuan CHEN ; Dongzhaoyang ZHANG ; Xin ZHONG ; Renqing JIANG ; Can CHEN ; Yijun JIA ; Zhaowen ZONG ; Bo HU
Journal of Army Medical University 2025;47(8):756-765
Objective To evaluate the resuscitation efficacy of novel lyophilized platelets(LP,thrombin-stimulated platelets)combined with lactated Ringer's(LR)solution in rabbits with hemorrhagic shock and seawater immersion.Methods Fifty rabbits were randomly assigned to 5 groups(Groups A,B,C,D and E,n=10).After all rabbits were anesthetized with 3%pentobarbital sodium at a dose of 1 mL/kg,soft tissue injury was inflicted in the left lower limb.The blood loss from the soft tissue injury was quantified after gauze hemostasis.The right lower limb was subjected to femoral artery catheterization,followed by blood withdrawal equivalent to 26%of the total blood volume of the rabbit.The rabbits were then vertically immersed in 3%artificial seawater,with the water level reaching above the xiphoid process,and were retrieved in 15 min later.Resuscitation strategies were applied to the rabbits from different group:Group A(no resuscitation),Group B(resuscitation with LR solution),Group C(resuscitation with LR solution and fresh platelets),Group D(resuscitation with LR solution and LP),and Group E(resuscitation with LR solution and novel LP).Coagulation function test,routine blood test,blood gas analysis,and thromboelastography were conducted at baseline and at 1,2 and 4 h after injury.Results The LP and rabbit model of hemorrhagic shock and seawater immersion were successfully prepared.At 1 h after injury,the mean arterial pressure(MAP)of Groups C,D and E(infused with platelet preparations)was significantly higher than that of Group A(without resuscitation,P<0.05);the lactate(Lac)content of Group C was obviously lower than that of Groups A and B(P<0.05);the base excess(BE)and blood urea nitrogen(BUN)levels of Groups C,D and E were notably lower than those of Groups A and B(P<0.05);and the prothrombin time(PT)of Group A was significantly longer than that of before injury(P<0.05).At 2 h after injury,the MAP of Groups C and D was significantly higher than that of Groups A and B,and that of Group E was notably higher than that of Group A(P<0.05);the Lac content of Groups C and E was obviously lower than that of Groups A and B,while that of Group D was also lower than that of Group A(P<0.05);the BE and BUN levels of Groups C,D and E were remarkably lower than those of Groups A and B(P<0.05);the maximum amplitude(MA)value of Group C was higher than that of Group A,while the value of Groups A and D at this time was significantly lower than the corresponding value before injury(P<0.05);and the activated partial clotting time(APTT)of Groups A and D was statistically longer than the corresponding baseline time(P<0.05).At 4 h after injury,the MAP of Groups C,D,and E was higher than that of Groups A and B,and that of Group B was higher than that of Group A(P<0.05);the Lac and BUN levels of Groups C,D,and E were lower than those of Groups A and B(P<0.05);the BE level of Groups C and D were lower than those of Groups A and B(P<0.05);the MA value of Groups B,C,and E were higher than those of Group A(P<0.05),while the MA value and APTT value of Groups A and D were significantly lower than their corresponding baseline values(P<0.05).Conclusion For rabbits with hemorrhagic shock and seawater immersion,the novel LP combined with LR solution can significantly increase the MAP level,reduce Lac content,and sustainably maintain blood clot firmness and coagulation function.
5.Construction of a preoperative prediction model for post-hepatectomy liver failure in patients with large hepatocellular carcinoma
Zhaowen ZHANG ; Xinyuan HU ; Zixiang CHEN ; Jiangming CHEN ; Xiaoping GENG ; Fubao LIU
Chinese Journal of General Surgery 2025;34(7):1390-1400
Background and Aims:Hepatocellular carcinoma(HCC)is the most prevalent type of liver malignancy,accounting for 80%of all primary liver cancer cases.Partial hepatectomy is widely considered to be the treatment of choice for HCC.However,post-hepatectomy liver failure(PHLF)is the most serious complication and the leading cause of perioperative death.Therefore,an accurate assessment of the risk of PHLF is particularly critical.Patients with large hepatocellular carcinoma have larger tumors(tumor diameter≥5 cm)and more resected liver tissue,and are more likely to develop PHLF.Previous studies have used various methods to assess the risk of PHLF,including liver function,Child-Pugh classification,model for end-stage liver disease,albumin-bilirubin(ALBI),and aspartate aminotransferase-to-platelet ratio index score.However,no model has been developed for data on hepatectomy for large HCC.Therefore,this study aims to analyze the risk factors of PHLF in HCC patients with large tumor and to construct a preoperative nomogram prediction model to guide and optimize clinical decision-making.Methods:The clinical data of 927 patients with large liver cancer who underwent radical hepatectomy in the First Affiliated Hospital of Anhui Medical University(721 cases,training cohort)and the Second Affiliated Hospital of Anhui Medical University(206 cases,validation cohort)from January 2018 to June 2023 were retrospectively collected.The patients'baseline data,laboratory examination,imaging data,and surgical information were collected.Univariate analysis combined with multivariate analysis was used to screen out the independent risk factors for inducing PHLF,and binary Logistic regression was used to construct a prediction model for PHLF.ROC,calibration,and clinical decision curves verified the model's performance.Results:There were no significant differences in all preoperative data between the training and validation cohorts(P>0.05).Grade B or C PHLF occurred in 192 of 927 patients(20.7%),including 8 patients with grade C PHLF.Univariate and multivariate Logistic regression analyses were used to determine the independent risk factors of PHLF,including tumor diameter,ALBI score,liver cirrhosis,vascular tumor thrombus,and intraoperative blood loss.These factors were included in the Logistic regression analysis,and a nomogram model was constructed to predict PHLF.The nomogram model was validated,and the C-index of the nomogram was 0.757.The ROC curve analysis of the prediction probability of the model showed that the AUC of the training set was 0.757(95%CI=0.703-0.811),and the AUC of the validation set was 0.779(95%CI=0.702-0.863).The validation showed that the model had good predictive ability.Conclusions:Tumor diameter,ALBI score,liver cirrhosis,vascular tumor thrombus,and intraoperative blood loss are independent risk factors for PHLF.The nomogram prediction model constructed in this study can accurately assess the risk of preoperative PHLF,which is helpful for better clinical management,reducing the occurrence of PHLF,and improving the postoperative prognosis of patients.
6.Efficiency of Rituximab for the treatment of the initial episode of steroid-sensitive nephrotic syndrome in children
Tingting YUAN ; Bingbing ZHU ; Yan LI ; Qianqian PENG ; Huandan YANG ; Na CHEN ; Zhaowen ZHONG ; Ruifeng ZHANG
Chinese Journal of Applied Clinical Pediatrics 2025;40(2):125-129
Objective:To evaluate the efficacy and safety of Rituximab (RTX) combined with short-term use of glucocorticoids in the treatment of the initial episode of steroid-sensitive nephrotic syndrome (SSNS).Methods:A retrospective case summary.A total of 30 children with SSNS treated in the Department of Intrarenal Rheumatism and Immunology, Xuzhou Children′s Hospital from December 2021 to March 2023, were enrolled in this study.They were given a standard dose of RTX (375 mg/m 2) and glucocorticoids for a short term.The patients were followed up for 1 year, and the general condition, changes in CD19 + B lymphocytes expression after RTX treatment, the relapse rate, the median relapse-free survival and adverse reactions of RTX were recorded.Kaplan-Meier method was used to analyze the relapse-free survival time. Results:CD19 + B lymphocytes were depleted 2 weeks after RTX treatment, and the median time required for CD19 + B lymphocytes reconstitution was 5 months after RTX treatment.The 1-year relapse-free rate was 90.00%, and the median relapse-free survival was 11 months.The glucocorticoid discontinuation rate was 93.33% in 3 months after RTX treatment, and 100% in 6 and 12 months after RTX treatment.Adverse reactions included infusion reactions in 7 cases (23.33%), neutropenia/leukopenia in 5 cases (16.67%), hypoimmunoglobulinemia in 10 cases (33.33%), infections in 4 cases.One case was complicated with severe influenza A virus infection after CD19 + B lymphocytes reconstruction. Conclusions:A standard dose of RTX can significantly reduce the relapse frequency, maintain long-term remission of proteinuria, and reduce the dose of glucocorticoids in patients at the initial episode of SSNS.Thus, it is worthy of clinical promotion.
7.Construction of a preoperative prediction model for post-hepatectomy liver failure in patients with large hepatocellular carcinoma
Zhaowen ZHANG ; Xinyuan HU ; Zixiang CHEN ; Jiangming CHEN ; Xiaoping GENG ; Fubao LIU
Chinese Journal of General Surgery 2025;34(7):1390-1400
Background and Aims:Hepatocellular carcinoma(HCC)is the most prevalent type of liver malignancy,accounting for 80%of all primary liver cancer cases.Partial hepatectomy is widely considered to be the treatment of choice for HCC.However,post-hepatectomy liver failure(PHLF)is the most serious complication and the leading cause of perioperative death.Therefore,an accurate assessment of the risk of PHLF is particularly critical.Patients with large hepatocellular carcinoma have larger tumors(tumor diameter≥5 cm)and more resected liver tissue,and are more likely to develop PHLF.Previous studies have used various methods to assess the risk of PHLF,including liver function,Child-Pugh classification,model for end-stage liver disease,albumin-bilirubin(ALBI),and aspartate aminotransferase-to-platelet ratio index score.However,no model has been developed for data on hepatectomy for large HCC.Therefore,this study aims to analyze the risk factors of PHLF in HCC patients with large tumor and to construct a preoperative nomogram prediction model to guide and optimize clinical decision-making.Methods:The clinical data of 927 patients with large liver cancer who underwent radical hepatectomy in the First Affiliated Hospital of Anhui Medical University(721 cases,training cohort)and the Second Affiliated Hospital of Anhui Medical University(206 cases,validation cohort)from January 2018 to June 2023 were retrospectively collected.The patients'baseline data,laboratory examination,imaging data,and surgical information were collected.Univariate analysis combined with multivariate analysis was used to screen out the independent risk factors for inducing PHLF,and binary Logistic regression was used to construct a prediction model for PHLF.ROC,calibration,and clinical decision curves verified the model's performance.Results:There were no significant differences in all preoperative data between the training and validation cohorts(P>0.05).Grade B or C PHLF occurred in 192 of 927 patients(20.7%),including 8 patients with grade C PHLF.Univariate and multivariate Logistic regression analyses were used to determine the independent risk factors of PHLF,including tumor diameter,ALBI score,liver cirrhosis,vascular tumor thrombus,and intraoperative blood loss.These factors were included in the Logistic regression analysis,and a nomogram model was constructed to predict PHLF.The nomogram model was validated,and the C-index of the nomogram was 0.757.The ROC curve analysis of the prediction probability of the model showed that the AUC of the training set was 0.757(95%CI=0.703-0.811),and the AUC of the validation set was 0.779(95%CI=0.702-0.863).The validation showed that the model had good predictive ability.Conclusions:Tumor diameter,ALBI score,liver cirrhosis,vascular tumor thrombus,and intraoperative blood loss are independent risk factors for PHLF.The nomogram prediction model constructed in this study can accurately assess the risk of preoperative PHLF,which is helpful for better clinical management,reducing the occurrence of PHLF,and improving the postoperative prognosis of patients.
8.Efficiency of Rituximab for the treatment of the initial episode of steroid-sensitive nephrotic syndrome in children
Tingting YUAN ; Bingbing ZHU ; Yan LI ; Qianqian PENG ; Huandan YANG ; Na CHEN ; Zhaowen ZHONG ; Ruifeng ZHANG
Chinese Journal of Applied Clinical Pediatrics 2025;40(2):125-129
Objective:To evaluate the efficacy and safety of Rituximab (RTX) combined with short-term use of glucocorticoids in the treatment of the initial episode of steroid-sensitive nephrotic syndrome (SSNS).Methods:A retrospective case summary.A total of 30 children with SSNS treated in the Department of Intrarenal Rheumatism and Immunology, Xuzhou Children′s Hospital from December 2021 to March 2023, were enrolled in this study.They were given a standard dose of RTX (375 mg/m 2) and glucocorticoids for a short term.The patients were followed up for 1 year, and the general condition, changes in CD19 + B lymphocytes expression after RTX treatment, the relapse rate, the median relapse-free survival and adverse reactions of RTX were recorded.Kaplan-Meier method was used to analyze the relapse-free survival time. Results:CD19 + B lymphocytes were depleted 2 weeks after RTX treatment, and the median time required for CD19 + B lymphocytes reconstitution was 5 months after RTX treatment.The 1-year relapse-free rate was 90.00%, and the median relapse-free survival was 11 months.The glucocorticoid discontinuation rate was 93.33% in 3 months after RTX treatment, and 100% in 6 and 12 months after RTX treatment.Adverse reactions included infusion reactions in 7 cases (23.33%), neutropenia/leukopenia in 5 cases (16.67%), hypoimmunoglobulinemia in 10 cases (33.33%), infections in 4 cases.One case was complicated with severe influenza A virus infection after CD19 + B lymphocytes reconstruction. Conclusions:A standard dose of RTX can significantly reduce the relapse frequency, maintain long-term remission of proteinuria, and reduce the dose of glucocorticoids in patients at the initial episode of SSNS.Thus, it is worthy of clinical promotion.
9.Identification of parrot roundworms and analysis of its mitochondrial genome
Na LI ; Zhaowen REN ; Yani WANG ; Pian ZHANG ; Tongbao SUN ; Ziguo YUAN ; Xi-Aohu WANG
Chinese Journal of Veterinary Science 2024;44(11):2379-2385
To identify the intestinal roundworms of Eclectus roratus and Psittacula alexandri from the Guangdong region by morphological analysis and molecular biology techniques,and to amplify the mitochondrial genome of parrot roundworms using PCR,as well as to carry out phylogenetic analyses based on the sequences of their mitochondrial genes.The results showed that the parrot roundworms identified in this identification belongs to Ascaridia.nymphii and its mitochondrial genome consists of 12 protein coding genes,22 tRNA genes,2 ribosomal RNA genes and two non-coding regions.Phylogenetic analyses showed that A.nymphii identified in this study is located in the same evolutionary branch as the previously reported Ascaridia sp.,is closely related and con-stitutes a separate clade of avian ascarids with pigeon roundworms and chicken roundworms.This study provides important data for the classification,molecular epidemiology and population genetic evolution of parrot roundworms.
10.Effects and mechanism of mechanical stretch promoting proliferation and inhibiting osteogenic and adipogenic differentiation of skeletal satellite cells in mice
Zhaowen GAO ; Hongyun LI ; Jianbin WU ; Shuxin ZHANG ; Jian YE ; Fengchun WU
Chinese Journal of Sports Medicine 2024;43(8):637-646
Objective To investigate the effect of mechanical stretch on the proliferation and differen-tiation of skeletal satellite cells in mice,and explore its mechanism.Methods The research was divid-ed into two parts.In the first part,all mice were not given osteogenic or adipogenic induction,and randomly divided into a control group,a mechanical stretch(MS)group,a solvent control group[Di-methyl sulfoxide(DMSO)+mechanical stretch],a Notch inhibitor(KY-02111)+mechanical stretch group and a Wnt inhibitor(FLI-06)+mechanical stretch group,with the aim to evaluate cell prolifera-tion and migration,and the effect of mechanical stretch on the Wnt and Notch signaling pathway.In the second part,all mice were randomly divided into 6 groups:a control group,an induced differentia-tion(ID)group,an induced differentiation+mechanical stretch(IDMS)group,an induced differentia-tion+DMSO+mechanical stretch(IDDMS)group,an induced differentiation+Notch inhibitor(KY-02111)+mechanical stretch(IDNMS)group,and an induced differentiation+Wnt inhibitor(FLI-06)+mechanical stretch(IDWMS)group.All groups except the control group were induced differenti-ation using osteoblastic or adipogenic medium,and the IDMS group was applied tensile stress to the cell basement membrane,with the IDDMS and IDNMS groups given DMSO and KY-02111 or FLI-06 with a final concentration of 5 μM into the culture medium,respectively.On the 7th and 14th days af-ter culture,cell proliferation was detected using CCK-8,while on the 14th day after culture,cell mi-gration was observed using the streak method.Alkaline phosphatase and oil red staining were per-formed on all cells.Moreover,the mRNA expression of osteogenic[alkaline phosphatase(ALP)and Runt-related transcription factor(Runx)-2]and adipogenic[fatty acid-binding protein(FABP)4 and li-poprteinlipase(LPL)]differentiation marker genes were quantitatively detected by using the real-time PCR.Meanwhile,while the expression of c-Myc and Cyclin B(CCNB)1 proteins was measured using Western blotting.Results Cell proliferation increased significantly in MS group compared with the con-trol group(P<0.01),while that of IDMMS and IDNMS groups decreased significantly compared with MS group.As to cell migration,MS group was significantly better than the control group(P<0.01),while there was a significant decrease in both IDMMS and IDNMS groups compared with MS group(P<0.01).According to ALP and oil red staining,a significant increase in ALP activity and lipid droplet secretion was observed in skeletal muscle satellite cells of ID groups compared with the control group,with the increase in IDMMS and IDNMS groups significantly greater than MS group.More-over,the results of real-time PCR showed that the expression of ALP and Runx-2mRNA in the osteo-genic induction group and the FABP4 and LPL mRNA in the adipogenic induction group increased sig-nificantly compared with the control group(P<0.05),with a significant decrease in the above measure-ments of IDMS group compared with ID group,but a significant increase in IDMMS and IDNMS groups compared with IDMS group(P<0.01).Meanwhile,according to Western blotting,the expres-sion of c-Myc and CCNB1proteins increased significantly in the skeletal muscle satellite cells of MS group compared with the control group(P<0.01),while that of IDMMS and IDNMS groups decreased significantly compared with MS group(P<0.01).Conclusion Mechanical stretch can promote the prolif-eration and migration of skeletal satellite cells in mice,but inhibit osteogenic and adipogenic differenti-ation.Its mechanism is related to the activation of Wnt and Notch signal pathways.

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