OBJECTIVES: To explore the molecular mechanism of Jiangzhi Huaban Decoction (JZHBD) for improving atherosclerosis through the "qi meridian-blood channels" pathway. METHODS: ApoE^-/- mouse models of atherosclerosis were established by high-fat diet feeding for 8 weeks, with C57BL/6 mice on a normal diet as the controls. Forty ApoE^-/- mouse models were randomized into model group, low-, medium-, and high-dose JZHBD treatment groups, and atorvastatin treatment group (n=8) for their respective treatments for 8 weeks. The changes in body weight and overall condition of the mice were monitored weekly. After the treatments, serum levels of TC, TG, HDL-C, LDL-C, TBA, ALT, and AST of the mice were measured, pathological changes in the liver and aortic root plaques were examined with HE staining, and lipid accumulation in the liver and aortic wall was assessed using Oil Red O staining. The core molecular mechanism was studied through transcriptomics, and the expressions of the key pathway proteins were confirmed using Western blotting and immunohistochemistry. RESULTS: Treatment with JZHBD significantly reduced blood lipid and total bile acid levels, improved liver function and hepatic steatosis, and decreased aortic lipid deposition and plaque area in the mouse models of atherosclerosis. Transcriptomic analysis suggested that the therapeutic mechanism of JZHBD involved reverse cholesterol transport, PPAR signaling, and the inflammatory pathways. In atherosclerotic mice, JZHBD treatment obviously up-regulated hepatic expressions of PPARγ, LXRα, ABCA1, ABCG1, and CYP7A1, down-regulated hepatic expressions of p-p65/p65, IL-6, IL1β in the liver, increased ABCG5 and ABCG8 expressions in the intestines, and decreased ICAM-1 and VCAM-1 expressions in the aortic plaques. CONCLUSIONS JZHBD improves atherosclerotic vascular damage and plaque formation possibly by regulating hepatic reverse cholesterol transport and inflammation via modulating the hepatic PPARγ/LXRα/NF-κB signaling pathway.
Animals ; Drugs, Chinese Herbal/therapeutic use* ; Mice, Inbred C57BL ; Plaque, Atherosclerotic/metabolism* ; Liver/metabolism* ; Mice ; Atherosclerosis/metabolism* ; Cholesterol/metabolism* ; PPAR gamma/metabolism* ; Male ; Diet, High-Fat ; Biological Transport

Background Per- and polyfluoroalkyl substances (PFAS) are a class of persistent organic pollutants widely used in various products, leading to population exposure and long-term accumulation. At present, there is a lack of research on the relationships between pre-pregnancy PFAS and menstrual characteristics among women undergoing assisted reproductive technology (ART) in China. Objective To explore the relationships between pre-pregnancy PFAS exposure among women undergoing ART and menstrual characteristics prior to assisted reproductive treatment. Methods This study employed a cross-sectional research design, recruiting women undergoing ART treatment at the Reproductive Clinic of the International Peace Maternity & Child Health Hospital, School of Medicine, Shanghai Jiao Tong University, from 2017 to 2020 as study participants. Ultra-high-performance liquid chromatography-tandem mass spectrometry was used to detect 42 types of PFAS in pre-pregnancy serum samples. Questionnaires were administered to collect information on demographic characteristics, lifestyle habits, and menstrual characteristics (average menstrual cycle length, average menstrual period length, menstrual irregularities, and menstrual bleeding volume) of women undergoing ART. Multiple linear regression, binary logistic regression, and multinomial logistic regression analyses were performed to investigate the relationships between individual PFAS exposure before pregnancy and menstrual characteristics among ART women. Additionally, weighted quantile sum (WQS) model was applied to analyze the association between PFAS mixtures and menstrual characteristics. Results In the pre-pregnancy serum samples of the study population, 15 PFAS were detected in more than 60% of the samples, including perfluoroheptanoic acid (PFHpA), perfluorooctanoic acid (PFOA), perfluorononanoic acid (PFNA), perfluorodecanoic acid (PFDA), perfluoroundecanoic acid (PFUnDA), perfluorododecanoic acid (PFDoDA), perfluorobutanesulfonic acid (PFBS), perfluorohexanesulfonic acid (PFHxS), perfluoroheptanesulfonic acid (PFHpS), perfluorooctanesulfonic acid (PFOS), 6:2 chlorinated polyfluorinated ether sulfonate (6:2 Cl-PFESA), 8:2 chlorinated polyfluorinated ether sulfonate (8:2 Cl-PFESA), perfluoro-2-propoxypropanoic acid (HFPO-DA), perfluoro-2-methoxyacetic acid (PFMOAA), and perfluoro-(3,5,7,9,11-pentaoxadodecanoic) acid (PFO5DoDA). Among them, PFOA had the highest median concentration of 9.160 ng·mL⁻¹. The single PFAS exposure analysis revealed a positive correlation between PFAS and irregular menstrual cycles. Specifically, for every natural-log unit (e) increase in PFOA, PFBS, or PFHxS level, the incidence of irregular menstrual cycles increased by 57%, 42%, or 39%, respectively. Most PFAS were positively correlated with the average number of menstrual cycle days, such as PFHpA (b=1.08, 95%CI: 0.11, 2.05), PFOA (b=1.69, 95%CI: 0.39, 3.00), PFBS (b=1.23, 95%CI: 0.25, 2.22), PFHxS (b=1.47, 95%CI: 0.61, 2.32), PFHpS (b=1.48, 95%CI: 0.35, 2.61), and 6:2 Cl-PFESA (b=0.90, 95%CI: 0.08, 1.72). Furthermore, levels of PFHpA (OR=1.39, 95%CI: 1.06, 1.82), PFOA (OR=1.58, 95%CI: 1.09, 2.30), PFBS (OR=1.37, 95%CI: 1.04, 1.80), PFHxS (OR=1.34, 95%CI: 1.05, 1.71), PFHpS (OR=1.53, 95%CI: 1.10, 2.14), and 6:2 Cl-PFESA (OR=1.34, 95%CI: 1.06, 1.70) were positively correlated with low menstrual blood volume, while PFOA (OR=0.40, 95%CI: 0.23, 0.71), PFHpS (OR=0.45, 95%CI: 0.29, 0.71), and HFPO-DA (OR=0.68, 95%CI: 0.48, 0.97) were negatively correlated with high menstrual blood volume. The mixed exposure model showed that PFAS mixtures were positively correlated with the average number of menstrual cycle days (b=1.60, 95%CI: 0.49, 2.71), irregular menstrual cycles (OR=1.77, 95%CI: 1.19, 2.63), and low menstrual blood volume (OR=1.59, 95%CI: 1.08, 2.35), but negatively correlated with high menstrual blood volume (OR=0.40, 95%CI: 0.22, 0.73). Conclusion Women undergoing ART in Shanghai are widely exposed to PFAS prior to conception. Exposure to PFAS before pregnancy may be related to menstrual characteristics among women seeking ART before undergoing fertility treatments, but additional data from larger populations are required to validate the findings of this study.

Antibody (Ab) humanization is critical to reduce immunogenicity and enhance efficacy in the preclinical phase of the development of therapeutic Abs originated from animal models. Computational suggestions have long been desired, but available tools focused on immunogenicity calculation of whole Ab sequences and sequence segments, missing the individual residue sites. This study introduces Site-specific Immunogenicity for Therapeutic Antibody (SITA), a novel computational framework that predicts B-cell immunogenicity score for not only the overall antibody, but also individual residues, based on a comprehensive set of amino acid descriptors characterizing physicochemical and spatial features for antibody structures. A transfer-learning-inspired framework was purposely adopted to overcome the scarcity of Ab-Ab structural complexes. On an independent testing dataset derived from 13 Ab-Ab structural complexes, SITA successfully predicted the epitope sites for Ab-Ab structures with a receiver operating characteristic (ROC)-area unver the ROC curve (AUC) of 0.85 and a precision-recall (PR)-AUC of 0.305 at the residue level. Furthermore, the SITA score can significantly distinguish immunogenicity levels of whole human Abs, therapeutic Abs and non-human-derived Abs. More importantly, analysis of an additional 25 therapeutic Abs revealed that over 70% of them were detected with decreased immunogenicity after modification compared to their parent variants. Among these, nearly 66% Abs successfully identified actual modification sites from the top five sites with the highest SITA scores, suggesting the ability of SITA scores for guide the humanization of antibody. Overall, these findings highlight the potential of SITA in optimizing immunogenicity assessments during the process of therapeutic antibody design.

Objective To investigate the expression of T cell subsets in the spleen of BTBR T+Itpr3tf autistic mouse at 4,8,and 12 weeks of age,and to determine the optimal age for studying the relationship between immune abnormalities and autism in BTBR autistic mice.Methods It randomly selected 5～6 male BTBR mouse at 4 weeks,8 weeks,and 12 weeks of age and C57BL/6J mouse of the same gender at corresponding ages for the three-box social interaction test,the self-grooming test,and the marble-burying test;Single cell suspensions were prepared from the spleens of mouse at 8 and 12 weeks of age,and flow cytometry was used to detect 8 subsets of T cells(TH 1,TH2,TH17,TC1,TC2,TC17,TFH,and Treg).Results Compared with C57BL/6J mouse of the same age,BTBR mouse at 4 weeks,8 weeks,and 12 weeks of age showed a decrease in social time(P＜0.001),an increase in grooming time(P＜0.01,P＜0.001),and an increase in the number of marbles buried(P＜0.01,P＜0.001)in BTBR mouse at 8 weeks and 12 weeks of age.As well,the expression of TH 1(P＜0.001),TH2(P＜0.01),TC 1(P＜0.05),TC2(P＜0.001),and TFH(P＜0.01)cells in 8-week-old BTBR mouse were significantly increased,while the expression of Treg(P＜0.001)cells were significantly decreased;The expression of TH 1(P＜0.01),TH2(P＜0.01),TH 17(P＜0.05),TC1(P＜0.01),TC2(P＜0.001),TC 17(P＜0.01),and TFH(P＜0.001)increased in 12-week-old BTBR mouse,while the expression of Treg(P＜0.05)cells decreased.At different age stages(P＜0.050)the ratio of TH 1/Treg and TC 1/Treg in 8-week-old BTBR mouse were significantly higher than those in 12 week old mouse,while the TC 17/Treg ratio decreased.Conclusions BTBR mouse at different developmental stages exhibit varying degrees of abnormal increase in Teff/Treg ratio.Based on result of behavioral test,it is recommended to use 8-week-old BTBR mice for research on autism and immune abnormalities.

Objective To investigate the expression of T cell subsets in the spleen of BTBR T+Itpr3tf autistic mouse at 4,8,and 12 weeks of age,and to determine the optimal age for studying the relationship between immune abnormalities and autism in BTBR autistic mice.Methods It randomly selected 5～6 male BTBR mouse at 4 weeks,8 weeks,and 12 weeks of age and C57BL/6J mouse of the same gender at corresponding ages for the three-box social interaction test,the self-grooming test,and the marble-burying test;Single cell suspensions were prepared from the spleens of mouse at 8 and 12 weeks of age,and flow cytometry was used to detect 8 subsets of T cells(TH 1,TH2,TH17,TC1,TC2,TC17,TFH,and Treg).Results Compared with C57BL/6J mouse of the same age,BTBR mouse at 4 weeks,8 weeks,and 12 weeks of age showed a decrease in social time(P＜0.001),an increase in grooming time(P＜0.01,P＜0.001),and an increase in the number of marbles buried(P＜0.01,P＜0.001)in BTBR mouse at 8 weeks and 12 weeks of age.As well,the expression of TH 1(P＜0.001),TH2(P＜0.01),TC 1(P＜0.05),TC2(P＜0.001),and TFH(P＜0.01)cells in 8-week-old BTBR mouse were significantly increased,while the expression of Treg(P＜0.001)cells were significantly decreased;The expression of TH 1(P＜0.01),TH2(P＜0.01),TH 17(P＜0.05),TC1(P＜0.01),TC2(P＜0.001),TC 17(P＜0.01),and TFH(P＜0.001)increased in 12-week-old BTBR mouse,while the expression of Treg(P＜0.05)cells decreased.At different age stages(P＜0.050)the ratio of TH 1/Treg and TC 1/Treg in 8-week-old BTBR mouse were significantly higher than those in 12 week old mouse,while the TC 17/Treg ratio decreased.Conclusions BTBR mouse at different developmental stages exhibit varying degrees of abnormal increase in Teff/Treg ratio.Based on result of behavioral test,it is recommended to use 8-week-old BTBR mice for research on autism and immune abnormalities.

Objective To explore the mechanism of TubA in the treatment of ulcerative colitis in mice,and to lay a foundation for the treatment strategy of ulcerative colitis.Methods Twenty C57BL/6 mice aged 6-8 weeks were randomly divided into 4 groups(n=5):the control group drank pure water every day,the model group and the treatment groups drank 3％dextran sulfate sodium(DSS)every day,and the treatment groups were injected with 10 mg/kg TubA and 20 mg/kg TubA every day from the third day,respectively.The weight changes of mice in all groups were recorded.Nine days later,the serum of mice was collected,and the expression levels of inflammatory factors IL-1β,IL-6 and IL-10 in serum were detected by ELISA.HE staining was used to observe the pathological changes of the mouse colon.The expression of myeloperoxidase(MPO)was detected by immunohistochemistry,the mRNA levels of inflammatory factors IL-1β,IL-6 and IL-10 were detected by RT-qPCR,and the expressions of GPX4 and FTH were detected by immunohistochemistry.The mRNA and protein expression levels of GPX4,GCLM,FTH,Nrf2,Keap1 and HO-1 in colon tissues were detected by RT-qPCR and Western blotting.Results Compared with the control group,the body weight and colon length of the model group decreased significantly.HE staining showed that inflammatory cells infiltrated the mucosa and submucosa of colon tissue,goblet cells were lost and crypt structure disordered and disappeared.Immunohistochemistry showed that the expression of MPO and FTH proteins were significantly increased,while the expression of GPX4 protein was significantly decreased(P＜0.05).The mRNA and protein expression levels of IL-1β and IL-6 were significantly increased,while the mRNA and protein expression levels of IL-10 were significantly decreased(P＜0.05).The mRNA and protein expression levels of FTH,Nrf2 and HO-1 were significantly increased,while the mRNA and protein expression levels of GPX4,GCLM and Keap1 were significantly decreased(P＜0.05).After TubA treatment,compared with the model group,all these changes mentioned above suppressed(P＜0.05).Conclusion TubA may reduce ulcerative colitis symptoms by inhibiting ferroptosis,providing new ideas for the treatment of ulcerative colitis.

Objective To investigate the biomechanical characteristics of the mandibular anterior teeth intrusion assisted by buccal mini-implants,so as to provide theoretical guidance for the clinical treatment of deep overbite and other conditions requiring intrusion of the mandibular anterior teeth.Methods A finite element model of implant screws,clear aligners(CAs)and mandibular complex including the mandibular dentition,periodontal ligament(PDL)and alveolar bone was constructed.The model was designed with 0.2 mm intrusion in the mandibular anterior teeth.Seven groups were set:without traction,30 g,50 g,100 g,150 g,200 g,and 250 g force groups.The stress and displacement of the teeth as well as the related stress distributions of PDL and CAs in each group after loading were analyzed.Results Without traction and under different traction forces,the mandibular anterior teeth tended to be tipped and intruded.With the increase of traction force,the increasing trend was manifested in the vertical displacement and labial displacement of the mandibular anterior teeth.When the traction force reached 200 g,the vertical displacement showed a significant increase,the subsequent increase then showed a tendency to flatten.The ratio of the labial tipping displacement to the intrusive displacement of the mandibular anterior teeth showed a decreasing trend with the increase of traction force.Within a reasonable force range,increasing traction force led to greater bodily intrusion of the anterior teeth.When the traction force was less than 150 g,the mandibular posterior teeth exhibited a tendency of extrusion.When the traction force of more than 150 g was applied,it showed a tendency of intrusion.The PDL stress was concentrated on the labial and cervical surface of the anterior teeth.Conclusions Intrusion efficiency of the mandibular anterior teeth has been effectively improved with CAs assisted by buccal mini-implants.Moreover,the traction force of the mini-implants has a significant impact on the movement of the mandibular anterior teeth.Based on a comprehensive analysis of factors such as the displacement trend of the mandibular dentition and the PDL stress,the optimal force value for buccal traction of the mini-implants to intrude the mandibular anterior teeth can be selected as 150-200 g force.

Objective To investigate the biomechanical characteristics of the mandibular anterior teeth intrusion assisted by buccal mini-implants,so as to provide theoretical guidance for the clinical treatment of deep overbite and other conditions requiring intrusion of the mandibular anterior teeth.Methods A finite element model of implant screws,clear aligners(CAs)and mandibular complex including the mandibular dentition,periodontal ligament(PDL)and alveolar bone was constructed.The model was designed with 0.2 mm intrusion in the mandibular anterior teeth.Seven groups were set:without traction,30 g,50 g,100 g,150 g,200 g,and 250 g force groups.The stress and displacement of the teeth as well as the related stress distributions of PDL and CAs in each group after loading were analyzed.Results Without traction and under different traction forces,the mandibular anterior teeth tended to be tipped and intruded.With the increase of traction force,the increasing trend was manifested in the vertical displacement and labial displacement of the mandibular anterior teeth.When the traction force reached 200 g,the vertical displacement showed a significant increase,the subsequent increase then showed a tendency to flatten.The ratio of the labial tipping displacement to the intrusive displacement of the mandibular anterior teeth showed a decreasing trend with the increase of traction force.Within a reasonable force range,increasing traction force led to greater bodily intrusion of the anterior teeth.When the traction force was less than 150 g,the mandibular posterior teeth exhibited a tendency of extrusion.When the traction force of more than 150 g was applied,it showed a tendency of intrusion.The PDL stress was concentrated on the labial and cervical surface of the anterior teeth.Conclusions Intrusion efficiency of the mandibular anterior teeth has been effectively improved with CAs assisted by buccal mini-implants.Moreover,the traction force of the mini-implants has a significant impact on the movement of the mandibular anterior teeth.Based on a comprehensive analysis of factors such as the displacement trend of the mandibular dentition and the PDL stress,the optimal force value for buccal traction of the mini-implants to intrude the mandibular anterior teeth can be selected as 150-200 g force.

Objective To investigate the occurrence and characteristics of Antineutrophil cytoplasmic antibody(ANCA)associated vasculitis induced by methimazole,and to provide references for clinical safe drug use.Methods Case reports of ANCA associated vasculitis induced by methimazole published in Wanfang,CNKI,PubMed,and Web of Science were searched from the inception to October 31 st,2023.Demographic characteristics,drug use,complications,treatment and outcome were analyzed using descriptive statistical method.Results A total of 14 patients from 14literature were included.There were 3 males and 11 females with ages ranging from 8 to 79 years,with a mean age of(47.79±23.47)years.Four patients developed symptoms within 1 year,nine patients developed symptoms from 2 to 12 years,and 1 patient developed symptoms 24 years after medication.ANCA associated vasculitis affected kidney in 5 patients,lung and skin in 5patients,vision in 2 patients and heart in 2 patients.All patients discontinued methimazole,2 patients improved spontaneously without treatment,1 patient improved after anti-infection,and all others received hormonal or immunosuppressive therapy.1 patient developed death,and all others improved or were cured after treatment.Conclusions ANCA-associated vasculitis is a rare adverse reactionof methimazole.Most patients have a long latency period before the onset of disease,mainly involving multiple organs such asskin,kidney,lung,and eyesight.Clinicians should pay attention to differentiate it from primary vasculitis and discontinue the drug as soon as possible.When serious organ damage occurs,glucocorticoids and immunosuppressants should be adminstered promptly to avoid aggravation of the disease and endangerment of life.

Antibody humanization is critical to reduce immunogenicity and enhance efficacy in the preclinical phase of the development of therapeutic antibodies originated from animal models.Computational suggestions have long been desired,but available tools focused on immunogenicity calculation of whole antibody sequences and sequence segments,missing the individual residue sites.This study introduces Site-specific Immunogenicity for Therapeutic Antibody(SITA),a novel computational framework that predicts B-cell immunogenicity score for not only the overall antibody,but also individual residues,based on a comprehensive set of amino acid descriptors characterizing physicochemical and spatial features for antibody structures.A transfer-learning-inspired framework was purposely adopted to overcome the scarcity of Antibody-Antibody structural complexes.On an independent testing dataset derived from 13 Antibody-Antibody structural complexes,SITA successfully predicted the epitope sites for Antibody-Antibody structures with a receiver operating characteristic(ROC)-area unver the ROC curve(AUC)of 0.85 and a precision-recall(PR)-AUC of 0.305 at the residue level.Furthermore,the SITA score can significantly distinguish immunogenicity levels of whole human antibodies,therapeutic antibodies and non-human-derived antibodies.More importantly,analysis of an additional 25 thera-peutic antibodies revealed that over 70％of them were detected with decreased immunogenicity after modification compared to their parent variants.Among these,nearly 66％antibodies successfully iden-tified actual modification sites from the top five sites with the highest SITA scores,suggesting the ability of SITA scores for guide the humanization of antibody.Overall,these findings highlight the potential of SITA in optimizing immunogenicity assessments during the process of therapeutic antibody design.