Objective: To investigate the effect of sirtuin 2(SIRT2) on the proliferation and migration of cardiac fibroblasts(CFs)in C57BL/6 mice under angiotensin II(Ang Ⅱ) stimulation. Methods : The hearts were taken from 1 to 2 days C57BL/6 milk mice. After cutting and digesting, CFs were extracted by different adhesion centrifugation. After CFs attachment, the cells were cultured under control medium and Ang Ⅱ(100 nmol/L) medium and treated using OE-SIRT2 plasmid to overexpression the SIRT2 gene. RT-qPCR was used to detect mRNA expression of SIRT2 proliferating cell nuclear antigen(PCNA), periostin(POSTN)and type Ⅰ collagen procollagen A1(Col1A1), Western blot assay was used to measure the protein expression levels of SIRT2, PCNA, POSTN and Col1A1, CCK-8 assay and EdU assay were used to evaluate CFs proliferation rate, Transwell experiment was used to assess CFs migration activity. Results: Compared with control group, Ang Ⅱ stimulation led to down-regulation of SIRT2 expression in CFs, increased collagen expression, and promoted CFs proliferation and migration. The expression of SIRT2 was up regulated in CFs treated with OE-SIRT2 plasmid under Ang Ⅱ stimulation, Col1A1, POSTN and PCNA expression was down regulated, and CFs proliferation and migration ability decreased. Conclusion Overexpression of SIRT2 can inhibit the proliferation and migration of CFs under Ang Ⅱ stimulation, indicating that SIRT2 may be a key regulatory point in the onset and progression of cardiac fibrosis.

Objective To construct curcumin pyrazole derivative by the reaction of diketone of curcumin and benzylhydrazine based on the above structure-activity relationship,and to explore its antioxidant activity to provide experimental basis for the development of curcumin antioxidant derivative.Methods Curcumin-N-substituted pyrazole derivative was synthesized from curcumin and benzylhydrazine.The structures of the derivative were confirmed by infrared spectroscopy(IR),nuclear magnetic resonance spectroscopy(1H-NMR,13C-NMR)and LC-MS.The antioxidant activity in vitro of the derivative was evaluated by determination of curcumin and its pyrazole derivative scavenging ability for 2,2-diphenyl-1-picrylhydrazyl(DPPH)free radical and 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid(ABTS)free radical.Results Curcumin pyrazole derivative was successfully synthesized.Curcumin and its pyrazole derivative showed good free radical scavenging effects in the range of 4.6-73.6,6.25-100 μg·mL-1,respectively,with a significant dose-effect relationship.The half-maximal inhibition(IC50)values of curcumin and its pyrazole derivatives determined by DPPH method were 14.24,40.37 μg·mL-1,respectively,while the IC50 values of curcumin and its pyrazole derivatives determined by ABTS method were 36.65,19.26 μg·mL-1,respectively.Conclusion The antioxidant activity of β-dione of curcumin was retained through the substitution of the pyrazole ring,and the curcumin pyrazole derivative deserves further investigation as a potential antioxidant.

Objective Based on the theory of five circuits and six qi,the characteristics of circuit-qi indicators of the date of birth of hospitalized patients with pneumonia of Shaoyin-disease syndrome were explored.Methods The data collection was conducted in 422 hospitalized patients with pneumonia of shaoyin-disease syndrome admitted to the Department of Classical Chinese Medicine,Fangcun Hospital of Guangdong Provincinal Hospital of Chinese Medicine from January 20,2012 to June 30,2022.And then statistical analysis was performed for circuit-qi indicators of the date of birth of the patients.Results The goodness of fit by chi-square test showed that there were statistically significant differences in the distribution of the heavenly-stem year,earthly-branch year,yearly circuit,recombinant yearly circuit,predominant qi,and sitian-zaiquan(circuit qi of the first and second half of a year)at birthdate of hospitalized patients with pneumonia of shaoyin-disease syndrome(P＜0.05 or P＜0.001).And the results indicated that a higher risk of suffering from pneumonia of shaoyin-disease syndrome existed in the population born in the heavenly-stem ji year and earthly-branch chou year,in the yearly circuit being deficient earth circuit,in the recombinant yearly circuit being wind,in the predominant qi being taiyin damp earth,and in the sitian-zaiquan being taiyin damp earth and taiyang cold-water.Conclusion There is a correlation between the incidence of pneumonia of shaoyin-disease syndrome and the circuit-qi indicators of the date of birth of the patients,and the pathogenesis of circuits and qi at birth date is probably related with yang deficiency of spleen and kidney,and cold interweaved with dampness.

Objective:To investigate the role of dopamine receptors in the central amygdala (CeA) in reduction of the anxiety level by propofol in a mouse model of post-traumatic stress disorder (PTSD).Methods:Fifty-six SPF healthy adult male C57BL/6 mice, aged 10 weeks, weighing 20-25 g, were divided into 7 groups ( n=8 each) using a random number table method: control group (C group), PTSD group (P group), PTSD+ propofol group (PP group), PTSD+ fat emulsion group (PF group), PTSD+ propofol+ normal saline group (PPN group), PTSD+ propofol+ dopamine receptor D1 (DRD1) antagonist group (PP+ DRD1-Ant group), and PTSD+ propofol+ DRD2 antagonist group (PP+ DRD2-Ant group). The PTSD model was developed by continuous plantar electric shock for 3 days. Propofol 120 mg/kg was intraperitoneally injected after successful establishment of the model in PP group, and the equal volume of fat emulsion was intraperitoneally injected in PF group. In PPN group, PP+ DRD1-Ant group and PP+ DRD2-Ant group, the equal volume of normal saline, DRD1 antagonist hydrochloride and DRD2 antagonist eticlopride hydrochloride were injected in bilateral CeA regions, respectively, 30 min later the efficacy of drugs reached the peak, and then propofol 120 mg/kg was intraperitoneally injected. The anxiety levels were measured at 4 h (T 1) and day 3 after propofol injection (T 2) by the open field test and elevated cross maze test. Results:Compared with C group, the time spent entering the open and central areas was significantly shortened at T 1, 2, and the number of entering the open and central areas was decreased at T 1, 2 in P group ( P<0.001). Compared with P group, the time spent entering the open and central areas was significantly prolonged at T 1, the number of entering the open and central areas was increased at T 1 ( P<0.001), and no significant change was found at T 2 in PP group ( P>0.05), and no significant change was found in the aforementioned parameters at T 1, 2 in PF group ( P>0.05). Compared with PPN group, the time spent entering the open and central areas was significantly shortened at T 1, and the number of entering the open and central areas was decreased at T 1 in PP+ DRD2-Ant group ( P<0.001), and no significant change was found at T 1 in PP+ DRD1-Ant group ( P>0.05). Conclusions:Activation of DRD2 in the CeA is involved in the process by which propofol reduces the anxiety level of mice with PTSD.

Aim To investigate the regulatory effect of morphine postconditioning in the LSG on remodeling after myocardial infarction. Methods SD rats were randomly divided into four groups: sham operation group (Sham), myocardial infarction group (MI), myocardial infarction + saline group (Control) and myocardial infarction + morphine postconditioning group (MI + Morphine) . The rat MI model was constructed by ligating the left anterior descending coronary artery, and then morphine was given to the LSG by percutaneous posterior approach. After four weeks, the changes of cardiac function in rats were detected by ultrasound. Masson staining was used to detect fibrosis changes; the expression of Collagen I and Collagen III protein was detected by Western blot. The mRNA expression of ANP and BNP was detected by RT-qPCR. The expression of JJLOR in LSG was detected by immunofluorescence. The concentration of catecholamine in plasma and myocardial tissue was detected by ELISA. Results Compared with the sham group, the cardiac function of the MI group was significantly impaired, the myocardial tissue showed significant fibrosis changes, and the concentration of catecholamine in plasma and myocardial tissue significantly increased. Compared with the control group, the MI + Morphine group reduced myocardial fibrosis collagen deposition in rats after MI, inhibited the expression of ANP and BNP in myocardial tissue, reduced the concentration of catecholamine, and improved the cardiac function of MI rats. Immunofluorescence results showed that JJLOR was expressed in LSG after MI and increased after morphine postconditioning. Conclusions This study shows that morphine postconditioning in the LSG has a protective effect on myocardial remodeling after myocardial infarction. The mechanism may be related to the activation of JJLOR in the LSG by morphine and the reduction of catecholamine release from sympathetic nerve endings.

Objective A quality control(QC)system based on the electronic portal imaging device(EPID)system was used to realize the Multi-Leaf Collimator(MLC)position verification and dose verification functions on Primus and VenusX accelerators.Methods The MLC positions were calculated by the maximum gradient method of gray values to evaluate the deviation.The dose of images acquired by EPID were reconstructed using the algorithm combining dose calibration and dose calculation.The dose data obtained by EPID and two-dimensional matrix(MapCheck/PTW)were compared with the dose calculated by Pinnacle/TiGRT TPS for γ passing rate analysis.Results The position error of VenusX MLC was less than 1 mm.The position error of Primus MLC was significantly reduced after being recalibrated under the instructions of EPID.For the dose reconstructed by EPID,the average γ passing rates of Primus were 98.86%and 91.39%under the criteria of 3%/3 mm,10%threshold and 2%/2 mm,10%threshold,respectively.The average γ passing rates of VenusX were 98.49%and 91.11%,respectively.Conclusion The EPID-based accelerator quality control system can improve the efficiency of accelerator quality control and reduce the workload of physicists.

A dose reconstruction algorithm for electrionic portal imaging device(EPID)based on calibration and calculation is developed.The raw data of EPID in continuous acquisition mode are corrected for dark field and gain,and the gray level features of bright field are used to determine the field boundary.Subsequently,MU calibration,off-axis calibration and field size calibration are performed on the EPID data,and dose reconstruction is carried out based on the calibrated superimposed flux and the Monte Carlo model of the linac head.Nine cases of IMRT plans are selected for verification and measurement using EPID and MapCheck separately,and the passing rates between the two tools are compared under different gamma criteria(3%/3 mm and 2%/2 mm).For a planned case,the average passing rates of multiple cases verified by MapCheck under the two criteria were 99.02%±1.28%and 90.84%±4.49%,and the average passing rates of the EPID reconstruction models were 98.86%±1.19%and 91.39%±4.80%.Compared with MapCheck,the EPID reconstruction algorithm based on calibration and calculation has no significant difference in the passing rate of IMRT plan verification(P＞0.05),which meets the clinical requirements of dose verification.

Objective:The effect of bone marrow soluble B cell maturation antigen (sBCMA) expression on the efficacy and side effects of chimeric antigen receptor (CAR) -modified T-cell-targeting B cell maturation antigen (BCMA) in patients with multiple myeloma (MM) .Methods:This study involved 29 patients with relapsed or refractory MM (RRMM) who received humanized anti-BCMA CAR-T cell clinical trials from January 2018 to December 2021. The expression of sBCMA in bone marrow before and after anti-BCMA CAR-T cell treatment was detected by flow cytometry and compared.Results:①Two months after BCMA CAR-T cell treatment, 20 patients (68.97%) achieved an overall response (OR), whereas nine patients had stable disease (SD) or miner emission (MR). ②The expression of sBCMA in the bone marrow of 20 patients with OR was higher before treatment than after [26 926 (18 215, 32 488) ng/L vs 9 968 (6 634, 11 459) ng/L; P<0.001]; no significant difference was observed in patients with MR and SD [41 187 (33 816, 47 046) ng/L vs. 33 954 (31 569, 36 256) ng/L; P=0.145]; sBCMA expression in patients with OR before CAR-T cell treatment was lower than in patients with MR and SD ( P=0.005). ③No significant linear correlation was found between the peak value of CAR-T cells and sBCMA expression in the bone marrow of all 29 patients with RRMM ( R2=0.035, P=0.330). ④No significant difference in sBCMA expression was found between grades 0-1 CRS group (13 patients) and grades 2-4 CRS group [16 patients; 32 045 (18 742, 40 801) ng/L vs 29 102 (24 679, 38 776) ng/L, P=0.879], nor between grade 0 ICANS group (22 patients) and grade 1-3 ICANS group [seven patients; 30 073 (19 375, 40 065) ng/L vs 33 816 (22 933, 43 459) ng/L, P=0.763]. Conclusion:sBCMA expression in the bone marrow is related to the efficacy of BCMA CAR-T cell therapy in patients with RRMM, but is not significantly correlated with the severity of adverse events. It may serve as a predictive biomarker for the efficacy of BCMA CAR-T cell therapy in these patients.

Objective To explore the mechanism of glycoprotein,granulocyte macrophage-colony stimulating factor(GM-CSF)targeted inhibitor combined with interleukin(IL)-23 targeted inhibitor in alleviating spinal fibrosis in mice with ankylosing spondylitis(AS).Methods Six healthy subjects(HC group)and six AS patients(AS group)were recruited,and their peripheral venous blood were collected,the serum levels of tumor necrosis factor-α(TNF-α),IL-23,IL-17 and GM-CSF were detected.AS mice model were established.A total of 30 mice were randomly divided into the Control group,the Model group,the IL-17A Inh group(positive control group),the IL-23 Inh group,and the GM-CSF Inh+IL-23 Inh group,with 6 mice in each group.The levels of TNF-α,IL-23,IL-17 and GM-CSF in serum of mice were detected by ELISA.Western blot was used to detect the levels of epithelial mesenchymal transition(EMT)markers E-cadherin,N-cadherin,snail and Vimentin in muscle/ligament tissues around spine,as well as the levels of receptor activator of nuclear factor-κB ligand(RANKL),osteoprotegerin(OPG)and alkaline phosphatase(ALP)in spinal bone tissues.Micro-CT was used to measure the volume of new bone and the mature bone in the left hind paw and spine(L5～6 vertebrae)of mice.Results Compared with the HC group,the levels of TNF-α,IL-23,IL-17 and GM-CSF in serum of patients in the AS group were increased(P<0.05).Compared with the Control group,the levels of TNF-α,IL-23,IL-17 and GM-CSF in serum of mice in the Model group were increased(P<0.05),the relative expression level of E-cadherin was down-regulated(P<0.05),while the relative expression levels of N-cadherin,snail and Vimentin in muscle/ligament tissues around spine were up-regulated(P<0.05),the relative expression level of RANKL in spinal bone tissues was up-regulated(P<0.05),and the volume of new bone in the left hind paw and L5～6 vertebrae of mice increased(P<0.05).Compared with the Model group,the levels of the above indexes in the GM-CSF Inh+IL-23 Inh group were reversed(P<0.05),while there was no significant difference in the above indexes in the IL-23 Inh group(P>0.05).Compared with the Control group,the relative expression levels of OPG and ALP in spinal bone tissues of mice in the Model group were up-regulated(P<0.05).Compared with the Model group,there was no significant difference in the above indexes in the GM-CSF Inh+IL-23 Inh group(P>0.05).Conclusion The combination therapy of GM-CSF targeted inhibitor and IL-23 targeted inhibitor can reduce the inflammation level,alleviate the fibrosis of muscle and ligament tissues around spine,inhibit the expression of RANKL in the spinal bone tissues,reduce new bone formation and pathological bone remodeling,and protect the activity of the spine.

Aim To investigate the effect of total glu-cosides of paeony on inflammatory injury and TLR4/NF-κB/NLRP3 pathway in autoimmune thyroiditis(AIT)rats.Methods The experiment was divided into control group,model group,total glucosides of pae-ony(TGP),TLR4 inhibitor group and TGP+TLR4 ag-onist group,with 10 animals in each group.Except for the control group,the rats in other groups were subcu-taneously injected with thyroglobulin and Freund's ad-juvant to induce the AIT rat model.After six weeks of administration,thyroid histopathological changes were observed using hematoxylin-eosin(HE)staining;ser-um levels of TPOAb,TgAb,TSH,T3,T4,TNF-α,INF-γ,IL-1 β and IL-1 β were detected by enzyme-linked immunosorbent assay(ELISA);TLR4/NF-κB/NLRP3 pathway mRNAs and proteins expression in thyroid tis-sues were detected by RT-qPCR and Western blot.Re-sults Compared with the control group,the thyroid follicular epithelium of rats was significantly damaged,and the serum levels of TPOAb,TgAb,TSH,T3,T4,TNF-α,INF-γ,IL-1 β and IL-1 β increased(P＜0.01).The expression of TLR4/NF-κB/NLRP3 path-way mRNAs and proteins increased in the model group(P＜0.01).Compared with the model group,the damage of thyroid follicular epithelium was alleviated,and the serum levels of TPOAb,TgAb,TSH,T3,T4,TNF-α,INF-γ,IL-1 β and IL-1 β were reduced(P＜0.01),the expression of TLR4/NF-κB/NLRP3 path-way mRNAs and proteins were down-regulated in the TGP group and TLR4 inhibitor group(P＜0.01).Compared with TGP group,the damage of thyroid follic-ular epithelium was aggravated,and the levels of serum TPOAb,TgAb,TSH,T3,T4,TNF-α,INF-γ,IL-1 β and IL-1 β were elevated(P＜0.05 or P＜0.01),the pro-tein expressions of TLR4/NF-κB/NLRP3 pathway mR-NAs and proteins were up-regulated in TGP+TLR4 ag-onist group(P＜0.05 or P＜0.01).Conclusions TGP may play a protective role in thyroid by inhibiting the TLR4/NF-κB/NLRP3 pathway and improving the inflammatory injury of thyroid tissues.