1.Effects of wogonin on joint inflammation in collagen-induced arthritis rats via the endoplasmic reticulum stress pathway
Yuru WANG ; Siyuan LI ; Ye XU ; Yumeng ZHANG ; Yang LIU ; Huiqin HAO
Chinese Journal of Tissue Engineering Research 2025;29(5):1026-1035
BACKGROUND:Rheumatoid arthritis is an inflammatory disease.Many studies have shown that wogonin has a good anti-inflammatory effect on rheumatoid arthritis,but its exact efficacy and specific mechanism of action remain to be clarified. OBJECTIVE:To investigate the mechanism of wogonin ameliorating joint inflammation by regulating endoplasmic reticulum stress pathway in rats with collagen-induced arthritis. METHODS:(1)At the animal level:Female Wistar rats were divided into healthy control group,arthritis model group and wogonin treatment group.Rat models of arthritis in the latter two groups were established by subcutaneous injection of bovine type Ⅱ collagen and adjuvant.In the wogonin group,wogonin was given by gavage for 28 consecutive days after modeling.During this period,the rats in each group were weighed,and arthritis score and ankle swelling were measured every 7 days.After the experiment,the pathological changes of the joint were observed,the mRNA and protein levels of endoplasmic reticulum stress pathway GRP78 and CHOP were detected by qRT-PCR,western blot,and immunohistochemistry.(2)At the cellular level,cell counting kit-8 was used to detect the cytotoxic effect of wogonin on fibroblast-like synoviocytes from rats with collagen-induced arthritis.The fibroblast-like synoviocytes induced by thapsigargin were treated with different concentrations of wogonin.The levels of interleukin-1β and tumor necrosis factor-α in the cell supernatant were detected by ELISA,and the intracellular reactive oxygen species in each group were determined by DCFH-DA probe method.The mRNA and protein levels of GRP78,IRE1α,XBP1s and CHOP were detected by qRT-PCR and western blot,respectively. RESULTS AND CONCLUSION:Compared with the healthy control group,arthritis index score and ankle swelling degree in the arthritis model group were increased(P<0.01),synovial hyperplasia,inflammatory cell infiltration,cartilage destruction and bone erosion were observed in pathological sections,and the mRNA and protein expressions of GRP78 and CHOP in the ankle were significantly increased(P<0.01),which were mainly located in synovial tissue and articular surface.Compared with the arthritis model group,the arthritis index score and ankle swelling degree in the wogonin treatment group were decreased(P<0.05),synovial hyperplasia and the number of inflammatory cells were decreased,cartilage destruction and bone erosion were alleviated,the mRNA and protein expression levels of GRP78 and CHOP in the ankle were decreased(P<0.05),particularly in synovial tissue and on the articular surface.There was no significant difference in body mass among the three groups(P>0.05).In the cell experiment,200 μmol/L wogonin significantly reduced the survival rate of fibroblast-like synoviocytes(P<0.01).Compared with the blank control group,the levels of interleukin-1β,tumor necrosis factor-α,content of reactive oxygen species,and mRNA and protein expression of GRP78,IRE1α,XBP1s,and CHOP in the thapsigargin group were significantly increased(P<0.05);compared with the thapsigargin group,50 and 100 μmol/L wogonin significantly reduced the levels of interleukin-1β and tumor necrosis factor-α in the cell supernatant(P<0.05,P<0.01),and 100 μmol/L wogonin significantly reduced the content of reactive oxygen species(P<0.01)and down-regulated the mRNA and protein expression levels of GRP78,IRE1α,XBP1s and CHOP(all P<0.05).These results suggest that wogonin can effectively alleviate joint inflammatory responses in rats with collagen-induced arthritis,and the endoplasmic reticulum stress pathway may be the key target of its intervention.
2.Material basis and action mechanism of drug-containing serum of Modified Erxian Pill inhibiting macrophage pyroptosis
Siyuan LI ; Yuru WANG ; Ye XU ; Di GUO ; Nan NAN ; Yang LIU ; Jie ZHAO ; Huiqin HAO
Chinese Journal of Tissue Engineering Research 2025;29(19):4029-4037
BACKGROUND:Our previous study found that Modified Erxian Pill could alleviate inflammation in collagen-induced arthritis rats,but its mechanism needs to be further verified. OBJECTIVE:To analyze the components absorbed in the blood of Modified Erxian Pill,and observe the effect of the drug-containing serum of Modified Erxian Pill on pyroptosis of J774A.1 macrophages. METHODS:(1)Analysis of components absorbed in the blood of Modified Erxian Pill:Ultra-high performance liquid chromatography-high resolution mass spectrometry was used to detect and identify Modified Erxian Pill and its components absorbed in the blood.(2)Effect of the drug-containing serum of Modified Erxian Pill on pyroptosis of J774A.1 macrophages:Molecular docking technology was used to initially verify the sesquiterpenoids and NLRP3 in components absorbed in the blood of Modified Erxian Pill.J774A.1 macrophages were randomly divided into blank control group,lipopolysaccharide+adenosine triphosphate group,and lipopolysaccharide+adenosine triphosphate+Modified Erxian Pill with low(2.5%),medium(5%),and high(10%)dose groups.The release of lactate dehydrogenase in the cell supernatant of each group was detected according to the kit instructions.The levels of interleukin-1β and interleukin-18 in cell supernatant were detected in each group by ELISA.The cell membrane damage was detected by Hoechst/PI staining.The expression levels of NLRP3,Caspase-1,GSDMD,and GSDMD-N protein in the cells of each group were detected by western blot assay. RESULTS AND CONCLUSION:(1)A total of 32 active components of Modified Erxian Pill were identified,and 21 components entered the blood.The main components into blood included a variety of sesquiterpenoids.(2)Molecular docking results showed that 3-O-Acetyl-13-deoxyphomenone,Incensol oxide,Atractylenolide III,Rupestonic acid,and 3,7-Dihydroxy-9,11-eremophiladien-8-one had good binding activity with NLRP3.(3)Compared with the blank control group,lactate dehydrogenase activity and the expression levels of interleukin-1β and interleukin-18 were significantly increased in cell supernatant of lipopolysaccharide+adenosine triphosphate group(P<0.001).Hoechst/PI staining showed that the number of PI-positive cells was significantly increased.After the intervention of lipopolysaccharide+adenosine triphosphate+Modified Erxian Pill group,all of them showed different degrees of reduction.(4)Compared with the blank control group,NLRP3,Caspase-1,GSDMD,and GSDMD-N protein expression levels were significantly increased in the lipopolysaccharide+adenosine triphosphate group(P<0.05).Compared with lipopolysaccharide+adenosine triphosphate group,the protein expressions of NLRP3,Caspase-1,GSDMD,and GSDMD-N were significantly decreased in the lipopolysaccharide+adenosine triphosphate+Modified Erxian Pill group(P<0.05),and had a certain dose dependence.These findings verify that the drug-containing serum of Modified Erxian Pill may inhibit the pyroptosis of J774A.1 macrophages by regulating the NLRP3/Caspase-1/GSDMD pathway.
3.Experience in Staged Differentiation and Treatment of Cirrhotic Ascites Based on the Theory of Tonification and Purging
Haihang DONG ; Yujie CAI ; Huiqin ZHANG ; Yan CHEN ; Yuying TU ; Dongling WANG ; Yinqiang ZHANG
Journal of Traditional Chinese Medicine 2025;66(5):532-536
It is believed that patients with cirrhotic ascites exhibit a pathological mechanism characterized by the decline of healthy qi and the accumulation of pathogenic factors. Clinically, treatment should be based on the theory of tonification and purging, with a staged approach distinguishing between the active phase and the remission phase. The balance between tonification and purging should be adjusted according to the progression of pathogenic and healthy actors. In the acute phase, purging should take precedence over tonification, using purging as a means of tonification to facilitate the flow of water and qi through the triple energizer. The severity of water retention, dampness, blood stasis, and heat should be carefully assessed to ensure thorough elimination of pathogenic factors while avoiding harm to healthy qi. Medication adjustments should be made once the pathogenic factors are significantly weakened. In the remission phase, an integrated approach combining both tonification and purging should be adopted, incorporating purging within tonification to clear residual pathogens and prevent recurrence. Concurrently, proactive treatment of the underlying disease is essential to achieve complete recovery and prevent the recurrence of ascites.
4.Efficiency and safety of haematopoietic stem cell collection in healthy donors
Rui HE ; Bangqiang ZHU ; Huiqin WEN ; Haijing WANG ; Maohong BIAN ; Yujie DIAO
Chinese Journal of Blood Transfusion 2025;38(2):209-213
[Objective] To explore the key factors affecting the efficiency and safety of hematopoietic stem cell apheresis. [Methods] The clinical data of 59 healthy donors who underwent allogeneic hematopoietic stem cell donation in the First Affiliated Hospital of Anhui Medical University from January 2021 to June 2024 were retrospectively analyzed. The number of CD34+ cells was used to evaluate the eligibility of stem cell collection. The effects of donor gender, age, patient weight, as well as the number of WBC, MNC, RBC, Hb, HCT, PLT, CD34+ cells, CD34+ percentage and instrument operating parameters on collection efficiency were analyzed. [Results] A total of 59 donors were enrolled, and 68 occasions of stem cell apheresis were performed, with a qualified collection rate of 56%. Donor gender, age, patient weight, total blood circulation volume, anticoagulant dosage, collection time, calcium gluconate dosage and RBC, Hb, HCT levels were not significantly correlated with the collection effect (P>0.05). Multivariate logistic regression analysis showed that the number of MNC cells, CD34+ cells and stem cell product volume were the key factors affecting the efficiency and safety. A total of 12 donors had mild adverse reactions during the collection process, and all of them were improved after treatment. [Conclusion] Optimizing apheresis strategy based on the three factors of MNC, WBC count and stem cell product volume on the day of collection will help to achieve high-quality collection and improve the success rate of transplantation.
5.Genetic detection for hereditary cancer syndrome among general population
Xinning CHEN ; Li ZHANG ; Li YU ; Huiqin JIANG ; Fei HUANG ; Chunyan ZHANG ; Baishen PAN ; Beili WANG ; Wei GUO
Chinese Journal of Clinical Medicine 2025;32(4):627-633
Objective To examine the significance of susceptible gene detection for hereditary cancer syndrome (HCS) among general population. Methods A total of 2 928 individuals undergoing routine health examinations in Healthcare Center of Zhongshan Hospital, Fudan University, from September 2021 to April 2024 were enrolled retrospectively. Next generation sequencing was employed to identify susceptible genes for HCS. American College of Medical Genetics and Genomics (ACMG) guideline was used to analyze the pathogenicity of variants. Clinical data, imagings, follow-up data were also collected. Results The overall mutation rate of HCS panel was 3.59% (105/2 928), with 0.61% (18/2 928) for MutY DNA glycosylase (MUTYH), 0.27% (8/2 928) for breast cancer susceptibility gene 1/2 (BRCA1/2) and 0.23% (7/2 928) for mismatch repair (MMR) genes. Conclusions Healthy individuals carrying tumor susceptible genes usually lack the relevant clinical phenotypes. Whether comprehensive testing needs to be carried out among healthy people remains to be further explored.
6.Relationship between eNOS gene polymorphism and main complications in premature infants
Xiaoyan Li ; Bing Li ; Jia' ; an Wang ; Xian Dong ; Huiqin Wang ; Haijuan Zhu ; Bin Zhang
Acta Universitatis Medicinalis Anhui 2025;60(4):719-724
Objective :
To explore the polymorphism of endothelial nitric oxide synthase(eNOS) gene in umbilical cord blood of preterm infants and its relationship with major complications in preterm infants.
Methods :
A total of 254 preterm infants(<37 weeks) who were hospitalized were selected as the study subjects. Umbilical cord blood was collected at delivery to determine the genotypes and alleles of eNOS gene at three loci: rs61722009, rs2070744,and rs1799983. Clinical data of the preterm infants were recorded, and the relationship between eNOS gene polymorphism and major complications in preterm infants was analyzed.
Results:
(1) The TC+CC genotype at locus rs2070744 was an independent risk factor for bronchopulmonary dysplasia(BPD) in preterm infants, with an OR(95%CI) of 1.266(1.017-1.577).(2) The GT+TT genotype at locus rs1799983 was an independent risk factor for retinopathy prematurity(ROP), with an OR(95%CI) of 1.184(1.008-1.391).(3) The AB+AA genotype at locus rs61722009 was also an independent risk factor for ROP,with an OR(95%CI) of 1.335(1.033-1. 726).(4) There was no significant relationship between gene polymorphism and the occurrence of respiratory distress syndrome( RDS) and periventricular-intraventricular hemorrhage( PIVH).
Conclusion
eNOS gene polymorphism is associated with the occurrence of BPD and ROP in preterm infants. The evaluation of e NOS gene polymorphism by umbilical cord blood measurement is helpful for the prevention and correct management of some serious complications.
7.Changes and clinical significance of SDF-1,MCP-1 and sCD44 levels in aqueous humor of patients with diabetic cataract
Xiaoyu QU ; Hongna ZHU ; Anle SU ; Huiqin LU ; Bing WANG
International Journal of Laboratory Medicine 2025;46(6):694-697,703
Objective To analyze the changes and clinical significance of aqueous humor stromal cell-de-rived factor-1(SDF-1),macrophage chemoattractant protein-1(MCP-1)and soluble adhesion molecule CD44(sCD44)in patients with diabetic cataract(DC).Methods A total of 80 patients with DC admitted to the hos-pital from January 2021 to January 2023 were selected as the DC group,and 40 patients with simple cataract during the same period were selected as the age-related cataract group.According to the stage of cataract,DC patients were divided into group A(incipient stage,32 cases),group B(intumescent stage,26 cases)and group C(mature stage and over mature stage,22 cases).According to the presence or absence of macular ede-ma after treatment,the patients were divided into occurrence group(20 cases)and non-occurrence group(60 cases).The levels of SDF-1,MCP-1 and sCD44 in each group were detected by enzyme-linked immunosorbent assay.The receiver operating characteristic curve and area under the curve(AUC)were used to analyze the value of SDF-1,MCP-1 and sCD44 levels in the diagnosis of DC.Results The levels of SDF-1,MCP-1 and sCD44 in the DC group were higher than those in the age-related cataract group(P<0.05),and the levels of SDF-1,MCP-1 and sCD44 in the A,B and C groups increased sequentially(P<0.05).The level of MCP-1 in the occurrence group was higher than that in the non-occurrence group(P<0.05).The AUC of MCP-1,sCD44 and SDF-1 in the diagnosis of DC was 0.869,and the diagnostic efficiency was better.Conclusion The changes of aqueous SDF-1,MCP-1 and sCD44 levels are related to the stage of cataract in DC patients.Dynam-ic monitoring of these three indexes,especially MCP-1,is helpful to judge the condition and prognosis of DC patients.
8.Machine vision and learning for evaluating different rancidity grades of Prunus mandshurica (Maxim.) Koehne
Yashun Wang ; Huirong Chen ; Jianting Gong ; Yang Cui ; Huiqin Zou ; Yonghong Yan
Journal of Traditional Chinese Medical Sciences 2025;2025(2):287-296
ObjectiveTo explore a rapid and accurate method for evaluating the quality of Prunus mandshurica (Maxim.) Koehne (P. mandshurica, Ku Xing Ren) during rancidity using machine vision and learning.MethodsSensory evaluation and chemometrics were used to classify P. mandshurica quality grades after rancidity. Chemical indicators of the P. mandshurica quality change were determined to verify the obtained grades and support the subsequent modeling. The International Commission on Illumination color space was used to extract the color features of the P. mandshurica. Discrimination and prediction models based on color features combined with multiple machine learning algorithms were established using 10-fold cross-validation and external test set validation.ResultsThe P. mandshurica rancidity samples were allocated to three quality grades. The Bayes net model based on powder color successfully identified the P. mandshurica at different grades with an accuracy of 88.89% and 100% using two validations, and the naive Bayes model based on section color achieved the same accuracy with an receiver operating characteristic area of 0.979. The instance-based k-nearest neighbors model based on powder color performed best in predicting the amygdalin content [R2 = 0.9801, mean absolute error (MAE) = 0.2071, root mean squared error (RMSE) = 0.4170], followed by the random committee model in predicting the acid value (R2 = 0.9580, MAE = 1.5121, RMSE = 1.9099) and the random forest model in predicting the peroxide value (R2 = 0.8857, MAE = 0.0027, RMSE = 0.0035).ConclusionThis study demonstrates that color digitization analysis is a potential method for rapidly evaluating the quality of P. mandshurica across the rancidity process, providing a new reference for the quality assessment of traditional Chinese medicines.
9.Analysis of the suspected cases of measles and rubella in Pudong New Area of Shanghai, 2013‒2022
Xiao WANG ; Aihua ZHANG ; Huiqin FU ; Yuying YANG ; Xiaoxian CUI ; Lipeng HAO ; Yanqiu ZHOU ; Lifeng PAN
Shanghai Journal of Preventive Medicine 2024;36(6):534-539
ObjectiveTo analyze the epidemic characteristics of measles and rubella in Pudong New Area of Shanghai from 2013 to 2022, and to provide data support for the elimination of measles and rubella. MethodsEnzyme linked immunosorbent assay was used to detect IgM antibodies in serum samples. The sequence of 630 nucleotides at the C-terminal of N gene of measles virus was amplified by reverse transcription-polymerase chain reaction and the phylogenic tree was constructed. ResultsA total of 1 529 suspected cases of measles were detected from 2013 to 2022, among which the positive rate of measles IgM antibody was 33.55% (513/1 529). The highest positive rate (20.73%) was from March to May , and the positive rate of rubella IgM antibody was 6.80% (104/1 529). The positive rate of both IgM was higher in males than that in females (P<0.05). The IgM against measles was mainly detected in 0‒ years old (63.16%, 96/152) and 20‒ years old (45.61%, 161/353). The IgM against rubella was mainly detected in 10‒20 years old (27.27%, 18/66). The IgM antibody could be detected more easily from 4 to 28 days after eruption, and the IgM antibody positive rate of measles/rubella from 2020 to 2022 was significantly lower than previous years (2013‒2019). There were 2 D8 genotype strains, and the rest were H1a gene subtypes. ConclusionThe positive rate of IgM antibodies against measles/rubella in Pudong New Area of Shanghai decreased significantly. People aged 0‒ years and 20‒ years old are more susceptible to measles, and rubella is concentrated in 10‒ years old. It is necessary to strengthen the vaccination of school-age children, in order to achieve the goal of eliminating measles. The age group with high risk of exposure should be checked for vaccination status to ensure the enhanced immunization, and the surveillance of imported measles cases should be strengthened.
10.Clinical evaluation of elderly patients with sarcopenia receiving Shenling baizhu powder combined with nutrition and exercise intervention
Huanhuan JIANG ; Dong CHEN ; Aishuang FU ; Yanlei GE ; Qiang WANG ; Shizi LI ; Huiqin SHEN ; Xia LI ; Yingcong WANG ; Yunfei CHEN
China Pharmacy 2024;35(21):2652-2657
OBJECTIVE To explore the safety, effectiveness, and cost-effectiveness of elderly patients with sarcopenia receiving Shenling baizhu powder combined with nutrition and exercise intervention, providing a reference for rational clinical drug use. METHODS A total of 237 elderly sarcopenia patients were randomly assigned to an observation group (118 cases) and a control group (119 cases). Both groups of patients received nutrition and exercise intervention; the observation group added Shenling baizhu powder (6 g each time, three times daily) on this basis. The safety, effectiveness, and cost-effectiveness of the two plans were compared after 3 months. RESULTS Both groups of patients completed the follow-up. Before intervention, no significant difference was observed in skeletal muscle index (SMI), grip strength, and 6-minute walk test (6MWT) speed between the two groups (P>0.05). After intervention, the grip strength of the patients in the observation group was significantly greater than that of the control group (25.05 kg vs. 23.18 kg, P<0.01); the treatment response rate of sarcopenia, SMI, and 6MWT speed were higher than those of the control group, butthe differences were not statistically significant (P>0.05). The adverse reaction/event rate of the patients in the observation group was lower than that of the control group (14.41% vs. 16.81%, P=0.611), but the difference was not statistically significant. Compared with the control group’s plan, the cost of the observation group’s plan was higher (981.25 yuan vs. 913.94 yuan), and the effect was better (effectiveness rate: 0.618 6 vs. 0.563 0), with an incremental cost-effectiveness ratio of 1 210.61 yuan. The results of the sensitivity analysis were consistent with the cost-effectiveness analysis results. CONCLUSION Elderly patients with sarcopenia who receive Shenling baizhu powder combined with nutrition and exercise intervention can significantly strengthen grip strength without increasing the incidence of adverse reactions/events. Compared with the control group plan, the observation group needs to spend an additional 1 210.61 yuan for each additional effective patient with sarcopenia.


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