ObjectiveTo systematically identify the chemical constituents of Xuefu Zhuyutang（XFZY） and quantitatively determine its main components， aiming to elucidate its pharmacodynamic material basis and provide a scientific foundation for improving its quality control standards. MethodsUltra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry（UPLC-Q-TOF-MS/MS） was employed for qualitative analysis of XFZY， and the identification of compounds was accomplished by comparing their retention times， secondary MS fragment ion information， 52 reference standards and relevant databases， followed by attribution of their herbal sources. A total of 22 representative compounds were screened out， and UPLC-quadrupole-linear ion trap mass spectrometry（UPLC-Q-TRAP-MS/MS） was applied for quantitative analysis of the compounds in the formula. ResultsA total of 77 compounds were identified in XFZY， including 31 flavonoids mainly derived from Aurantii Fructus， Glycyrrhizae Radix et Rhizoma， Persicae Semen， Carthami Flos， Bupleuri Radix， Paeoniae Radix Rubra and Achyranthis Bidentatae Radix， 24 terpenoids mainly derived from Platycodonis Radix， Glycyrrhizae Radix et Rhizoma， Paeoniae Radix Rubra and Rehmanniae Radix， 9 phenylpropanoids and their derivatives mainly derived from Chuanxiong Rhizoma， Angelicae Sinensis Radix and Rehmanniae Radix， 4 phenolic acids mainly derived from Chuanxiong Rhizoma， Angelicae Sinensis Radix and Paeoniae Radix Rubra， 3 saccharides mainly derived from Rehmanniae Radix and Achyranthis Bidentatae Radix， and 6 other compounds mainly derived from Persicae Semen， Rehmanniae Radix and Angelicae Sinensis Radix. The results of quantitative analysis showed that the contents of protocatechuic acid， hydroxypaeoniflorin， amygdalin， vanillic acid， paeoniflorin， liquiritin apioside， liquiritin， isoquercitrin， naringin， cosmosiin， hesperidin， neohesperidin， isoliquiritin， liquiritigenin， naringenin， benzoylpaeoniflorin， hesperetin， isoliquiritigenin， formononetin， glycyrrhizic acid， nobiletin and ligustilide in XFZY were determined to be 0.12， 1.57， 54.53， 0.29， 36.17， 4.29， 4.84， 0.09， 46.67， 0.04， 3.44， 31.95， 0.82， 0.10， 0.11， 0.43， 0.07， 0.03， 0.01， 8.24， 0.13， 1.81 mg·g^-1. ConclusionThe qualitative method established in this study enables rapid and sensitive analysis of the chemical constituents in XFZY. Among the identified compounds， 52 are confirmed by reference standards， ensuring the accuracy of identification. The quantitative analysis of 22 key components provides a reliable experimental basis for the pharmacodynamic material basis research and quality control standard improvement of XFZY.

Objective To compare the differences in intraoperative radiation exposure between leadless and transvenous pacemaker implantation. Methods Cumulative dose (CD), dose area product (DAP), and fluoroscopy time during procedure were recorded and analyzed in 21 patients with leadless pacemaker implantation (Micra group), 55 patients with transvenous single-chamber pacemaker implantation (VVI group), and 216 patients with transvenous dual-chamber pacemaker implantation (DDD group). Results The fluoroscopy times of the Micra group, VVI group, and DDD group were 5.0 ± 1.9, 4.8 ± 1.4, and 7.6 ± 1.9 min, respectively (P < 0.001). Their CD values were 203.5 ± 76.1, 147.0 ± 41.0, and 249.6 ± 58.2 mGy, respectively (P < 0.001). Their DAP values were 18.6 ± 7.1, 13.4 ± 3.9, and 22.6 ± 5.6 Gy·cm², respectively (P < 0.001). Compared with the VVI group, the Micra group had similar fluoroscopy time (P=0.813) but higher CD (P=0.010) and DAP values (P = 0.012). Compared with the DDD group, the Micra group had reduced fluoroscopy time (P < 0.001), CD value (P = 0.033), and DAP value (P = 0.047). Conclusion Leadless pacemaker implantation is associated with increased radiation exposure compared to transvenous single-chamber pacemaker implantation. However, it offers a significant advantage in reducing radiation exposure for both medical staff and patients compared to transvenous dual-chamber pacemaker implantation.

This study established the mouse podocyte clone-5 (MPC5) with Smad3 knockout and studied the effect of transforming growth factor-beta 1 (TGF-β1) on the dedifferentiation of the MPC5 cells with Smad3 knockout, aiming to provide a cell tool for studying the role of Smad3 in mouse podocytes. The single-guide RNA (sgRNA) sequence targeting Smad3 was designed according to the principles of CRISPR/Cas9 design. The pX458-Smad3 vector was constructed and introduced into competent cells, and then the vector was extracted and used to transfect MPC5 cells. The successfully transfected cells were sorted by a flow cytometer. After single-cell clone expansion, PCR amplification of sequences adjacent to the edition site of Smad3 and sequencing were performed to identify potential cells with gene knockout. Western blotting was employed to verify the knockout efficiency of Smad3. Finally, the effect of Smad3 knockout on TGF-β1-induced dedifferentiation of MPC5 cells was analyzed by reverse transcription-polymerase chain reacting (RT-PCR), Western blotting, and the immunofluorescence method. The sgRNA was designed to target the fifth exon of Smad3. EGFP expression was observed 24 h after transfection of the pX458-Smad3 plasmid into MPC5 cells, with the transfection efficiency of 0.1% as determined by flow cytometry. From the transfected cells, 21 cell clones were obtained through flow cytometric sorting and single-cell clone expansion. PCR amplification and sequencing of the region around the sgRNA target site in Smad3 identified two cell clones with biallelic frameshift mutations. Western blotting results confirmed the absence of Smad3 expression in these clones, indicating successful establishment of the MPC5 cell line with Smad3 knockout. In normal MPC5 cells, TGF-β1 stimulation promoted the expression of fibrosis-related genes fibronectin and Col1a1 (collagen I) and inhibited the expression of the podocyte marker proteins synaptopodin and podocin, which suggested epithelial-mesenchymal transition and podocyte injury. However, in the two MPC5 cell lines with Smad3 knockout, TGF-β1-induced expression of epithelial-mesenchymal transition markers was significantly suppressed. The MPC5 cell lines with Smad3 knockout that were constructed by CRISPR/Cas9 provide a valuable cell model for functional studies of Smad3 protein and highlight the critical role of Smad3 in cell dedifferentiation.
Animals ; Smad3 Protein/genetics* ; CRISPR-Cas Systems/genetics* ; Mice ; Podocytes/metabolism* ; Transforming Growth Factor beta1/pharmacology* ; Cell Line ; Gene Knockout Techniques ; RNA, Guide, CRISPR-Cas Systems/genetics*

Objective To investigate the correlation between serum Angiopoietin-like protein 4(ANGPTL4)and soluble bone marrow cell-like transcription factor-1(sTLT-1)levels and cognitive function and prognosis in elderly patients with vascular dementia(VD).Methods A total of 92 elderly patients with vascular dementia admitted to Hebei Yanda Hospital from June 2020 to June 2022 were selected as the study group.They were divided into mild(n=30),moderate(n=36)and severe groups(n=26)according to the minimum mental state examination(MMSE)score,and 92 patients with healthy physical examination during the same period were selected as the control group.According to the prognosis of patients,the cognitive dysfunction was divided into grade Ⅰ～Ⅱ and grade Ⅲ.ELISA method was used to detect the levels of serum ANGPTL4 and sTLT-1.Pearson and Spearman methods were used to analyze the correlation between serum ANGPTL4 and sTLT-1 and MOCA scores.Logistic regression analysis was used to analyze the influencing factors of the prognostic grade Ⅲ of elderly patients with vascular dementia.The receiver operating characteristic(ROC)curve was plotted to analyze the predictive value of serum ANGPTL4 and sTLT-1 levels in the prognostic grade Ⅲ in elderly patients with vascular dementia.Results The serum ANGPTL4 level(987.57±53.25 pg/ml)in the study group was lower than that in the control group(1 108.35±62.13 pg/ml),and the serum sTLT-1(68.01±5.15 pg/ml)level was higher than that in the control group(50.12±4.57 pg/ml),with significant differences(t=14.158,24.922,all P＜0.05).Serum ANGPTL4 levels and MOCA scores were decreased sequentially in the mild,moderate and severe groups(F=33.495,66.617),while serum sTLT-1 level was increased sequentially(F=66.718),and the differences were statistically significant(all P＜0.05),respective.Pearson analysis showed that serum ANGPTL4 was negatively correlated with sTLT-1(r=-0.621,P＜0.05).Spearman analysis showed serum levels of ANGPTL4 and sTLT-1 were positively and negatively correlated with MoCA scores,respectively(r=0.545,-0.557,all P＜0.05).The serum ANGPTL4 level(953.45±51.16 pg/ml)in the prognostic patients of grade Ⅲ was lower than that of grade Ⅰ～Ⅱ(1 005.76±54.27 pg/ml),and the serum level of sTLT-1(73.14±5.40 pg/ml)was higher than that of grade Ⅰ～Ⅱ(65.28±5.02 pg/ml),with significant differences(t=4.490,6.967,all P＜0.05).Logistic regression analysis showed low expression of ANGPTL4[OR(95％CI):5.089(1.833～14.129)]and high expression of sTLT-1[OR(95％CI):4.258(1.739～10.428)]were risk factors affecting the prognosis of elderly patients with vascular dementia(P＜0.05).According to the ROC curve,the combined prediction of two for the grade Ⅲ prognosis of elderly vascular dementia patients was better than the individual prediction of ANGPTL4 and sTLT-1(Z=2.135,3.268,all P＜0.05).Conclusion Serum ANGPTL4 level was decreased but sTLT-1 level was increased in elderly patients with vascular dementia,and ANGPTL4 and sTLT-1 were closely related to cognitive function and prognosis.

Objective To analyze the expression levels of serum microRNA(miR)-211 and miR-202 in patients with Alzheimer's disease and their correlation with cognitive function,anxiety and depression.Methods A total of 90 patients with Alzheimer's disease admitted to Hebei Yanda Hospital from March 2019 to March 2022 were selected as the research group.According to the Clinical Dementia Rating(CDR)score,the patients were grouped into mild group(n=24),moderate group(n=48)and severe group(n=18).Another 90 healthy individuals who underwent physical examination were collected as the control group.The expression levels of miR-211 and miR-202 in serum were compared.Pearson method and Spearman method were used to analyze serum miR-211 and miR-202 and their correlation with cognitive function,anxiety and depression.Logistic regression analysis was used to analyze the influencing factors of Alzheimer's disease.Results The expression levels of serum miR-211(0.59±0.16,1.01±0.31)and miR-202(0.35±0.10,1.00±0.32)were significantly reduced in the research group and control group,with significant differences(t=11.422,18.393,all P<0.05).Serum miR-211(0.73±0.21,0.62±0.17,0.32±0.08)expression levels,miR-202(0.51±0.15,0.33±0.10,0.19±0.04)expression levels,mini-mental state examination(MMSE)score(22.54±1.41 score,19.35±1.01 score,16.23±1.00 score)and Montreal cognitive assessment(MoCA)score(25.35±2.60 score,18.59±1.32 score,16.59±1.24 score)in the mild,moderate and severe groups gradually decreased,and the differences were statistically significant(F=32.006,46.917,163.048,163.703,all P<0.05).Compared with mild group,the serum miR-211,miR-202,MMSE and MoCA scores of severe group and moderate group were reduced,and the differences were statistically significant(t=3.685～25.375,all P<0.05).The mild,moderate and severe groups had a gradual increase in Hamilton anxiety scale(HAMA)score(12.34±1.27 score,20.59±2.09 score and 31.29±2.19 score)and Hamilton depression scale(HAMD)score(14.35±2.13 score,23.89±2.20 score and 35.35±1.21 score),and the differences were statistically significant(F=496.059,553.939,all P<0.05).According to Pearson correlation analysis,miR-211 was positively correlated with miR-202(r=0.651,P<0.05).According to Spearman correlation analysis,miR-211 and miR-202 were positively correlated with MMSE and MoCA(r=0.539～0.585,all P<0.05)and negatively correlated with HAMA and HAMD(r=-0.651～-0.539,all P<0.05).Logistic regression analysis showed that the low expression of miR-211[OR(95%CI):5.321(1.648～17.180)]and miR-202[OR(95%CI):3.158(1.989～5.012)]were risk factors for Alzheimer's disease(P<0.05).Conclusion The serum expression levels of miR-211 and miR-202 in patients with Alzheimer's disease were reduced,indicating miR-211 and miR-202 were closely related to cognitive function,anxiety and depression.

Objective To investigate the current situation of achievement transformation in tertiary medical hospitals in Shanghai and propose countermeasures for the existing problems to enhance the effectiveness of achievement transformation.Methods A questionnaire survey was done on the transformation and management of scientific research achievements in 47 terti-ary hospitals in Shanghai.Meanwhile,interviews were carried out among the managers and researchers from these hospitals.Re-sults In the past three years,the rate of transformation achievements in the hospitals was only 2.8%.In the achievement trans-formation existed such problems as weak awareness of scientific researchers,low patent quality,lack of full-time managers,and inflexible management mode.It was also believed that there is a need to improve main responsibilities,achievement management,system establishment,personnel training,resource sharing,department coordination and other related aspects.Conclusion The rate of achievement transformation in tertiary medical institutions in Shanghai is at a lower level.There are numerous problems and difficulties in the transformation.Therefore,urgent efficient countermeasures are needed to promote the transformation of a-chievement.

Objective Establish a standardized evaluation system for medical faults,and provide theoretical basis for medical institutions and related industries to evaluate the illegality of medical behaviors.Methods Based on a litera-ture review,the medical fault assessment system was initially constructed,and then a research group was estab-lished to use Delphi method to invite 31 experts to evaluate the importance and feasibility of each article of the medi-cal fault assessment system and put forward suggestions for modification.Results The effective recovery rates of the two rounds of expert consultation were 83.9％and 96.8％,the expert authority coefficient was 0.902 and 0.887,and the Kendall's W test of all levels differences were statistically significant(P＜0.001).The medical fault assess-ment system finally constructed includes 5 first-level items including practicing medicine according to law,informed notification,diagnosis and treatment technology,medical record documents and hospital management,as well as 10 second-level items,20 third-level items and 47 fourth-level items.The mean values of importance and feasibili-ty scores of all articles were greater than 4,standard deviations were less than 1,and coefficients of variation were less than 0.2.Conclusion The medical fault standardized evaluation system is scientific,reliable,innovative and appli-cable.

Objective Establish a standardized evaluation system for medical faults,and provide theoretical basis for medical institutions and related industries to evaluate the illegality of medical behaviors.Methods Based on a litera-ture review,the medical fault assessment system was initially constructed,and then a research group was estab-lished to use Delphi method to invite 31 experts to evaluate the importance and feasibility of each article of the medi-cal fault assessment system and put forward suggestions for modification.Results The effective recovery rates of the two rounds of expert consultation were 83.9％and 96.8％,the expert authority coefficient was 0.902 and 0.887,and the Kendall's W test of all levels differences were statistically significant(P＜0.001).The medical fault assess-ment system finally constructed includes 5 first-level items including practicing medicine according to law,informed notification,diagnosis and treatment technology,medical record documents and hospital management,as well as 10 second-level items,20 third-level items and 47 fourth-level items.The mean values of importance and feasibili-ty scores of all articles were greater than 4,standard deviations were less than 1,and coefficients of variation were less than 0.2.Conclusion The medical fault standardized evaluation system is scientific,reliable,innovative and appli-cable.

Objective Establish a standardized evaluation system for medical faults,and provide theoretical basis for medical institutions and related industries to evaluate the illegality of medical behaviors.Methods Based on a litera-ture review,the medical fault assessment system was initially constructed,and then a research group was estab-lished to use Delphi method to invite 31 experts to evaluate the importance and feasibility of each article of the medi-cal fault assessment system and put forward suggestions for modification.Results The effective recovery rates of the two rounds of expert consultation were 83.9％and 96.8％,the expert authority coefficient was 0.902 and 0.887,and the Kendall's W test of all levels differences were statistically significant(P＜0.001).The medical fault assess-ment system finally constructed includes 5 first-level items including practicing medicine according to law,informed notification,diagnosis and treatment technology,medical record documents and hospital management,as well as 10 second-level items,20 third-level items and 47 fourth-level items.The mean values of importance and feasibili-ty scores of all articles were greater than 4,standard deviations were less than 1,and coefficients of variation were less than 0.2.Conclusion The medical fault standardized evaluation system is scientific,reliable,innovative and appli-cable.

Objective Establish a standardized evaluation system for medical faults,and provide theoretical basis for medical institutions and related industries to evaluate the illegality of medical behaviors.Methods Based on a litera-ture review,the medical fault assessment system was initially constructed,and then a research group was estab-lished to use Delphi method to invite 31 experts to evaluate the importance and feasibility of each article of the medi-cal fault assessment system and put forward suggestions for modification.Results The effective recovery rates of the two rounds of expert consultation were 83.9％and 96.8％,the expert authority coefficient was 0.902 and 0.887,and the Kendall's W test of all levels differences were statistically significant(P＜0.001).The medical fault assess-ment system finally constructed includes 5 first-level items including practicing medicine according to law,informed notification,diagnosis and treatment technology,medical record documents and hospital management,as well as 10 second-level items,20 third-level items and 47 fourth-level items.The mean values of importance and feasibili-ty scores of all articles were greater than 4,standard deviations were less than 1,and coefficients of variation were less than 0.2.Conclusion The medical fault standardized evaluation system is scientific,reliable,innovative and appli-cable.