Objective:To perform RHD gene detection on a blood sample with serological weak D phenotype.Methods:A specimen received by the People's Hospital of Zhijin County was serologically identified by the microcolumn gel method and saline method.RHD gene detection was conducted by the PCR-SSP method,and the full sequence determination of the 10 exons amplified was performed.The sequencing results were compared with the ISBT database to determine the genotype.Bioinformatics tool was used to predict the functional damage of mutant proteins,and Alphafold-3 was used for tertiary structural modeling of wild-type and mutant RhD proteins,and the structures of the two proteins were compared and analyzed to explore the reasons why mutations lead to weak serological manifestations.Results:The patient's genotype was identified as RHD*DV.5/RHD*01N.01 heterozygote,with the complete deletion of RHD genes on one chromosome,unable to express the D antigen.On the other chromosome,a G＞A mutation occurred at the 697th base of the 5th exon,resulting in a partial D phenotype.This mutation causes internal hydrogen bond changes at the 233 position of RhD protein,resulting in a change in the conformation of the protein,affecting binding to the corresponding antibody.Conclusion:The patient is a heterozygous mutant individual with RHD*DV.5/RHD*01N.01,exhibiting a partial D phenotype serologically.This variation is extremely rare and has been scarcely reported globally.

Objective:To investigate the feasibility of deep learning reconstruction (DLR) algorithm combined with a dual-low protocol (low radiation dose and low contrast medium dose) for thoracoabdominal aortic CT angiography (CTA).Methods:This cross-sectional study prospectively enrolled 56 patients suspected of aortic diseases who underwent aortic CTA at China-Japan Friendship Hospital from June 2023 to June 2024. All patients were randomly divided into two groups: Group A (28 cases) underwent CTA with a tube voltage of 100 kVp, automatic tube current modulation (noise index=10), and a contrast agent dose of 80 ml (flow rate 5 ml/s), with images reconstructed using the three-dimensional adaptive iterative dose reduction algorithm (AIDR). Group B (28 cases) underwent CTA with a tube voltage of 80 kVp, automatic tube current modulation (noise index=25), and a contrast agent dose of 40 ml (flow rate 3.5 ml/s), with images reconstructed using either the deep learning reconstruction algorithm-Advanced intelligent Clear-IQ Engine (AiCE subgroup) or the AIDR (AIDR subgroup). Two physicians evaluated the image quality of the three groups subjectively and objectively. Objective evaluation metrics included CT values, image noise (SD), signal-to-noise ratio (SNR), and contrast-to-noise ratio (CNR) at the ascending aorta, carina-level descending aorta, celiac trunk-origin abdominal aorta, and common iliac bifurcation abdominal aorta carina. Subjective evaluation metrics included image quality and noise scores. Comparisons among the three datasets (Group A, AiCE subgroup, AIDR subgroup) were performed using one-way ANOVA or the Kruskal-Wallis test, with appropriate post-hoc tests for pairwise comparisons.Results:No significant differences were observed in CT values of the ascending aorta, descending aorta, and abdominal aorta between Group A and the AiCE subgroup or the AIDR subgroup ( P0.05). However, significant overall differences were found in SD, SNR, and CNR values for the ascending aorta, descending aorta, and abdominal aorta ( P0.05). Pairwise comparisons revealed that, except for no significant differences in SD, SNR, and CNR values of the ascending and descending aorta between Group A and the AiCE subgroup, and no significant difference in SNR values of the ascending and abdominal aorta between Group A and the AIDR subgroup ( P0.05), all other intergroup comparisons showed statistically significant differences ( P0.05). Significant overall differences were also observed in image quality and noise scores between Group A and the AiCE and AIDR subgroups ( P0.05). Except for no significant differences in image quality and noise scores between Group A and the AiCE subgroup ( P0.05), all other pairwise comparisons showed statistically significant differences ( P0.05). Conclusions:The application of deep learning reconstruction algorithm combined with a dual-low protocol in thoracoabdominal aortic CTA can reduce radiation dose and contrast agent dose while maintaining diagnostic image quality, demonstrating significant clinical value for widespread adoption.

Natural products are an important source for innovative drugs,but unclear molecular targets and mechanisms limit their further development and application.The authors proposed a new method for the target identification of natural products based on proteolysis-targeting chimera(PROTAC)technology and quantitative proteomics,and established the targeted degradomics(TGDO) technology for the identification of weak-affinity tar-gets.This article summarizes the standardized workflow and the application of TGDO for target identification of natural products.

Aim To explore the mechanism of Jiawei Shaofu Zhuyu decoction in antagonizing endometriosis fibrosis by regulating mitophagy.Methods After the animal model was constructed,the syndrome was evalu-ated by general condition,organ water content and ther-mal imaging.The curative effect was evaluated by the weight of ectopic focus and the degree of adhesion.The pathological changes were compared using HE stai-ning,transmission electron microscopy,Masson and Sir-ius red staining.The expression of PINK1 and Parkin was detected by immunohistochemistry.The expression of mRNA and protein was determined by qPCR and Western blot,and the level of serum ROS was detected by ELISA.Results The autonomic activity of model mice was weakened,the water content of organs rose,and the temperature of limbs and lower abdomen was reduced by thermal imaging.HE staining showed obvi-ous hyperplasia of ectopic epithelium and glands.Transmission electron microscopy showed mitochondrial and endoplasmic reticulum structure damage,and nor-mal autophagy structure disappeared.Masson and Siri-us red staining showed increased collagen deposition;immunohistochemistry showed decreased expression of PINK1 and Parkin in ectopic foci.qPCR and Western blot showed that the expression of PINK1,Parkin,Bec-lin1,LC3 mRNA and protein in ectopic foci of model mice decreased,the expression of p62 mRNA and pro-tein increased,and serum ROS increased.The syn-drome performance of model mice was improved after the intervention of Jiawei Shaofu Zhuyu decoction;the inflammatory infiltration of ectopic foci was relieved,the morphology of mitochondria and endoplasmic retic-ulum was restored,and normal autophagy structure ap-peared.The degree of collagen deposition and fibrosis was reduced;the mRNA and protein expression of PINK1,Parkin,Beclin1 and LC3 increased.The ex-pression of p62 mRNA and protein decreased,and the level of ROS decreased.Conclusions Jiawei Shaofu Zhuyu decoction can improve the fibrosis of ectopic le-sions in mice with endometriosis of cold-dampness sta-sis syndrome,which may be related to the regulation of mitophagy.

Natural products are an important source for innovative drugs,but unclear molecular targets and mechanisms limit their further development and application.The authors proposed a new method for the target identification of natural products based on proteolysis-targeting chimera(PROTAC)technology and quantitative proteomics,and established the targeted degradomics(TGDO) technology for the identification of weak-affinity tar-gets.This article summarizes the standardized workflow and the application of TGDO for target identification of natural products.

Aim To explore the mechanism of Jiawei Shaofu Zhuyu decoction in antagonizing endometriosis fibrosis by regulating mitophagy.Methods After the animal model was constructed,the syndrome was evalu-ated by general condition,organ water content and ther-mal imaging.The curative effect was evaluated by the weight of ectopic focus and the degree of adhesion.The pathological changes were compared using HE stai-ning,transmission electron microscopy,Masson and Sir-ius red staining.The expression of PINK1 and Parkin was detected by immunohistochemistry.The expression of mRNA and protein was determined by qPCR and Western blot,and the level of serum ROS was detected by ELISA.Results The autonomic activity of model mice was weakened,the water content of organs rose,and the temperature of limbs and lower abdomen was reduced by thermal imaging.HE staining showed obvi-ous hyperplasia of ectopic epithelium and glands.Transmission electron microscopy showed mitochondrial and endoplasmic reticulum structure damage,and nor-mal autophagy structure disappeared.Masson and Siri-us red staining showed increased collagen deposition;immunohistochemistry showed decreased expression of PINK1 and Parkin in ectopic foci.qPCR and Western blot showed that the expression of PINK1,Parkin,Bec-lin1,LC3 mRNA and protein in ectopic foci of model mice decreased,the expression of p62 mRNA and pro-tein increased,and serum ROS increased.The syn-drome performance of model mice was improved after the intervention of Jiawei Shaofu Zhuyu decoction;the inflammatory infiltration of ectopic foci was relieved,the morphology of mitochondria and endoplasmic retic-ulum was restored,and normal autophagy structure ap-peared.The degree of collagen deposition and fibrosis was reduced;the mRNA and protein expression of PINK1,Parkin,Beclin1 and LC3 increased.The ex-pression of p62 mRNA and protein decreased,and the level of ROS decreased.Conclusions Jiawei Shaofu Zhuyu decoction can improve the fibrosis of ectopic le-sions in mice with endometriosis of cold-dampness sta-sis syndrome,which may be related to the regulation of mitophagy.

BACKGROUND:Buqi Huoxue Compounds have shown significant clinical effects on the treatment of ischemic stroke due to qi deficiency and phlegm stasis,but its specific mechanism of action needs to be further clarified.OBJECTIVE:To observe the effects of Buqi Huoxue Compounds on the neurological function,p-Akt and serum exosome expression in a rat model of cerebral ischemia-reperfusion injury.METHODS:Forty adult male SPF Sprague-Dawley rats,6 weeks old,were randomly divided into sham operation group,model group,Buqi Huoxue Compounds group,Buqi Huoxue Compounds+GW4869 group,with 10 rats in each group.In the latter three groups,a rat model of cerebral ischemia-reperfusion injury was established using thread technique.The sham operation group was given incision and separation without inserting a suture.The Buqi Huoxue Compounds group was intragastrically given Buqi Huoxue Compounds(6.49 g/kg,administered three times a day)2 hours after reperfusion;the GW4869+Buqi Huoxue Compounds group was intragastrically given Buqi Huoxue Compounds(6.49 g/kg,administered three times a day)2 hours after reperfusion and intraperitoneally given GW4869[2.5 mg/(kg·d)]2 hours before gavage with 3 days after modeling.Both the sham operation group and model group received equal amounts of saline via gavage,three times a day,for 7 consecutive days.Neurological function,cerebral infarct volume,brain tissue morphology,characterization of serum exosome,p-Akt in the cortical area and CD63 and TSG101 in serum exosome were detected after 7 days of administration.RESULTS AND CONCLUSION:(1)After modeling,compared with the sham operation group,the neurological function scores of the model group,Buqi Huoxue Compounds group,and Buqi Huoxue Compounds+GW4869 group were significantly increased(P＜0.01 or P＜0.05).After 7 days of intervention,the neurological function scores of the Buqi Huoxue Compounds group were significantly lower than those of the model group and Buqi Huoxue Compounds+GW4869 group(all P＜0.01).(2)The results of 2,3,5-triphenyltetrazolium chloride staining showed that cerebral infarct foci were observed in the model group,Buqi Huoxue Compounds group,and Buqi Huoxue Compounds+GW4869 group,and the cerebral infarct volume in the Buqi Huoxue Compounds group was smaller than that in the model group and the Buqi Huoxue Compounds+GW4869 group(P＜0.01).(3)The results of hematoxylin-eosin staining showed that the morphological and structural abnormalities of nerve cells in the Buqi Huoxue Compounds group and Buqi Huoxue Compounds+GW4869 group were less severe than those in the model group,but some cells still exhibited cytoplasmic agglutination and pyknosis in the Buqi Huoxue Compounds group and Buqi Huoxue Compounds+GW4869 group.(4)NanoSight analysis showed that the diameters of the isolated particles ranged from 60 nm to 300 nm,consistent with the characteristic size of exosomes.(5)Western blot results showed that the expression of p-Akt in the infarct zone and expression of CD63 and TSG101 in serum exosomes were significantly lower in the model group compared with the sham operation group(P＜0.05 or P＜0.01).Compared with the model group,the expression of p-Akt in the infarct zone and expression of CD63 and TSG101 in serum exosomes were significantly higher in the Buqi Huoxue Compounds group than in the model group(P＜0.05 or P＜0.01).However,the p-Akt expression in the infarct zone and CD63 expression in serum exosomes decreased significantly in the Buqi Huoxue Compounds+GW4869 group(P＜0.05),while TSG101 expression decreased without significant difference after the addition of GW4869.(6)To conclude,Buqi Huoxue Compounds attenuate cerebral ischemia-reperfusion injury and increase the expression of p-Akt in rats by promoting exosome secretion.

Isoflavones which mainly distributed in leguminous plants have plenty of health benefits.Isoflavone synthase(IFS)is a membrane-associated cytochrome P450 enzyme(CYP450)which carries out the unique aryl-ring migration and hydroxylation.So far,few crystal structures of plant P450s have been obtained.We determined the crystal structure of IFS from Medicago truncatula at 1.9 ? by MAD method using a selenomethionine substituted crystal and conducted molecular docking and mutagenesis study.The structure of IFS complexed with imidazole exhibits the helix Ⅰa-loop-helix Ⅰβ motif which cor-responds to helix Ⅰ of other P450s.Compared with structures of common P450s,IFS/imidazole structure contains an extra domain,i.e.,the γ-domain.The structure reveals a homodimer in which the γ-domain of one molecule interacts with the β-domain of another.The plane of heme group makes an angle of ap-proximately 40° with the helix Ⅰa-loop-helix Ⅰβ motif.Molecular docking combined with mutagenesis study suggested that Trp-128 and Asp-300 might play important roles in substrate binding and recogni-tion.Phe-301,Ser-303 and Gly-305 from the helix Ⅰa-loop-helix Ⅰβ motif may play important roles in the aryl-ring migration.These novel structural features reveal insights into the unique reaction mechanism of IFS and provide a basis for engineering IFS in leguminous crops for health purpose.

Objective:To screen inflammatory markers that can be used for the diagnosis and intervention evaluation of patients with chronic kidney disease(CKD)with depression,and to systematically study the therapeutic effect and safety of modified Chaihu Longgu Muli decoction in patients with CKD with depression.Methods:This study was a prospective study,a total of 120 patients with CKD who were diagnosed and treated in the Department of Nephrology,Chongqing Traditional Chinese Medicine Hospital from April 2023 to October 2024 were included.They were divided into CKD with depression(CKD-D)group and CKD without depression(CKD-N)group according to the diagnostic criteria by random number table method.The results of routine laboratory tests were collected,and the severity of depressive disorder was evaluated by Hamilton Depression Rating Scale-17(HAMD-17).The levels of interleukin-18(IL-18),interferon-γ(IFN-γ),β-thromboglobulin(β-TG),platlet factor-4(PF-4),eosinophil chemotactic factor(Eotaxin),soluble tumor necrosis factor receptor-2(sTNFR-2)and CD40 ligand(CD40L)in serum were quantitatively evaluated by liquid chip technology.Pearson correlation analysis was used to analyze the correlation between HAMD-17 scores and inflammatory markers with statistical differences in CKD patients with depression at different stages.The patients in CKD-D group were randomly divided into control group and treatment group.The control group was given basic treatment of CKD,while the treatment group was treated with modified Chaihu Longgu Muli decoction on the basis of the control group.After 8 weeks of continuous intervention,the clinical effective rate,the changes of effective inflammatory markers and the occurrence of adverse reactions were compared between the two groups.Results:In the CKD-D group and the CKD-N group,the difference of HAMD-17 score,serum phosphorus(P),serum creatinine(Scr),estimated glomerular filtration rate(eGFR),albumin(ALB),serum iron(Fe3+),C-reactive protein(CRP),IL-18,IFN-γ,sTNFR-2 indicators were statistically significant(P＜0.05).Multivariate Logistic regression analysis showed that in addition to HAMD-17 score(OR=1.259,P=0.006),SCr(OR=1.748,P=0.003),eGFR(OR=1.354,P=0.005),serum IL-18(OR=0.924,P=0.011)and IFN-γ(OR=0.859,P=0.031)levels were also independent influencing factors for CKD patients with depression.Pearson correlation analysis showed that there was a significant positive correlation between HAMD-17 score and IL-18,IFN-γ,sTNFR-2 and CD40L.The total clinical effective rate of the treatment group was higher than that of the control group,and the serum levels of IL-18 and IFN-γ in the treatment group were lower than those in the control group,the differences were statistically significant(P＜0.05),and no significant difference in the incidence of adverse reactions between the two groups(P＞0.05).Conclusion:IL-18 and IFN-γ can be used as effective serum markers for the diagnosis and treatment of depressive disorders in CKD patients.At the same time,the changes of serum levels of IL-18 and IFN-γ can be used to evaluate the severity of symptoms in CKD patients with depression at different stages to a certain extent.Modified Chaihu Longgu Muli decoction may improve CKD combined with depressive symptoms and improve the quality of life of patients by down-regulating the level of inflammatory factors.

Isoflavones which mainly distributed in leguminous plants have plenty of health benefits.Isoflavone synthase(IFS)is a membrane-associated cytochrome P450 enzyme(CYP450)which carries out the unique aryl-ring migration and hydroxylation.So far,few crystal structures of plant P450s have been obtained.We determined the crystal structure of IFS from Medicago truncatula at 1.9 ? by MAD method using a selenomethionine substituted crystal and conducted molecular docking and mutagenesis study.The structure of IFS complexed with imidazole exhibits the helix Ⅰa-loop-helix Ⅰβ motif which cor-responds to helix Ⅰ of other P450s.Compared with structures of common P450s,IFS/imidazole structure contains an extra domain,i.e.,the γ-domain.The structure reveals a homodimer in which the γ-domain of one molecule interacts with the β-domain of another.The plane of heme group makes an angle of ap-proximately 40° with the helix Ⅰa-loop-helix Ⅰβ motif.Molecular docking combined with mutagenesis study suggested that Trp-128 and Asp-300 might play important roles in substrate binding and recogni-tion.Phe-301,Ser-303 and Gly-305 from the helix Ⅰa-loop-helix Ⅰβ motif may play important roles in the aryl-ring migration.These novel structural features reveal insights into the unique reaction mechanism of IFS and provide a basis for engineering IFS in leguminous crops for health purpose.