Dendrobium moniliforme (D. moniliforme) is a traditional medicinal herb widely cultivated in Asia. Flavonoids, one of the largest groups of secondary metabolites in plants, are significant medicinal components in Dendrobium species. Several subgroups of R2R3-MYB proteins have been validated to directly regulate flavonoid biosynthesis. Using PacBio sequencing technology, we assembled a high-quality chromosome-level D. moniliforme genome with a total length of 1.20 Gb and a contig N50 of 3.97 Mb. The BUSCO assessment of genome annotation was 91.4%. By integrating the genome and transcriptome, we identified biosynthesis pathway enzyme genes related to flavonoids, polysaccharides, carotenoids, and alkaloids. A total of 90 R2R3-MYBs were identified in D. moniliforme and classified into 21 subgroups. Studies on the functions of R2R3-MYB transcription factors revealed that R2R3-MYB in SG6 can up-regulate flavonoid biosynthesis. Various validation experiments, including subcellular localization, transient overexpression, UPLC-MS/MS, HPLC, yeast one-hybrid, and dual-luciferase assays, demonstrated that DMYB69 directly up-regulates the expression of enzyme genes involved in flavonoid biosynthesis, increasing the content of flavonoids such as anthocyanin, flavone, and flavonol. Additionally, DMYB44 was shown to directly up-regulate the expression of carotenoid biosynthesis enzyme genes, thereby increasing carotenoid content. This study provides an essential genome resource and theoretical basis for molecular breeding research in D. moniliforme.

This study investigated the role of the nuclear factor of activated T cells c3 (NFATc3) in vascular smooth muscle cells (VSMCs) during aortic aneurysm and dissection (AAD) progression and the underlying molecular mechanisms. Cytoplasmic and nuclear NFATc3 levels were elevated in human and mouse AAD. VSMC-NFATc3 deletion reduced thoracic AAD (TAAD) and abdominal aortic aneurysm (AAA) progression in mice, contrary to VSMC-NFATc3 overexpression. VSMC-NFATc3 deletion reduced extracellular matrix (ECM) degradation and maintained the VSMC contractile phenotype. Nuclear NFATc3 targeted and transcriptionally upregulated matrix metalloproteinase 9 (MMP9) and MMP2, promoting ECM degradation and AAD development. NFATc3 promoted VSMC phenotypic switching by binding to eukaryotic elongation factor 2 (eEF2) and inhibiting its phosphorylation in the VSMC cytoplasm. Restoring eEF2 reversed the beneficial effects in VSMC-specific NFATc3-knockout mice. Cabamiquine-targets eEF2 and inhibits protein synthesis-inhibited AAD development and progression in VSMC-NFATc3-overexpressing mice. VSMC-NFATc3 promoted VSMC switch and ECM degradation while exacerbating AAD development, making it a novel potential therapeutic target for preventing and treating AAD.

ObjectiveTo investigate the potential mechanism of Liangxue Tuizi Formula （凉血退紫方， LXTZF） in treating Henoch-Schönlein Purpura （HSP） by examining its regulatory effect on neutrophil extracellular trap （NETs） dysregulation via the rapidly accelerated fibrosarcoma kinase （RAF）/mitogen-activated protein kinase （MEK）/extracellular signal-regulated kinase （ERK） signaling pathway. MethodsSeventy Wistar rats were randomly allocated into a blank control group （n=14） and a modeling group （n=56）. Rats in the modelling group underwent an eight-week modelling period to establish HSP rat models with blood-heat syndrome via modified ovalbumin （OVA） induction method combined with oral administration of heat-property Chinese herbal medicine. Fifty successfully modeled rats were subsequently randomly divided into five groups （n=10 per group）， model group， compound glycyrrhizin group， LXTZF group， RAF inhibitor group， and LXTZF + RAF agonist group. Additionally， 10 rats were selected from the original blank control group for the final experiment. From the 11th week of modelling， rats in the blank control group and the model group received 1 ml/（100 g·d） ultrapure water via oral administration， in addition to 0.5 ml/（kg·d） 0.9% sodium chloride solution via intraperitoneal injection. The LXTZF group and the compound glycyrrhizin group received 7.5 g/（kg·d） LXTZF granule suspension via gavage， 13.5 mg/（kg·d） compound glycyrrhizin suspension via gavage， respectively. The RAF inhibitor group received 1 mg/（kg·d） GW5074 suspension via intraperitoneal injection and ultrapure water via oral administration； the LXTZF + RAF agonist group received 7.5 g/（kg·d） LXTZF granule suspension via gavage and 1 mg/（kg·d） paclitaxel suspension via intraperitoneal injection. All administrations were performed once daily for 4 weeks. After intervention， skin tissue histopathology was examined by hematoxylin and eosin （H&E） staining， immunoglobulin A （IgA） deposition was assessed via immunofluorescence， serum levels of neutrophil elastase （NE）， tumor necrosis factor-α （TNF-α）， and vascular cell adhesion molecule-1 （VCAM-1） were measured using enzyme-linked immunosorbent assay （ELISA）， serum myeloperoxidase （MPO） level was determined by a colorimetric assay； the mRNA expression levels of RAF， MEK， and ERK in skin tissue were detected by real-time quantitative polymerase chain reaction （RT-qPCR）； and the protein expression of RAF， MEK， ERK， as well as phosphorylated MEK （p-MEK） and phosphorylated ERK （p-ERK）， were analyzed by Western Blot. ResultsSkin tissue in the blank control group rats remained normal， whereas the model group exhibited neutrophil infiltration and haemorrhage with red blood cell rupture. In all drug intervention groups， neutrophil infiltration and haemorrhagic exudation reduced markedly， with LXTZF group demonstrating the most pronounced improvement. Compared with the blank control group， rats in the model group exhibited enhanced IgA fluorescence intensity in skin tissue， elevated serum levels of NE， MPO， TNF-α and VCAM-1， increased mRNA expression of RAF， MEK， ERK1 and ERK2， as well as heightened RAF protein levels and p-MEK/MEK and p-ERK/ERK ratios （P＜0.05）. Compared with the model group， the drug intervention groups exhibited reduced IgA fluorescence intensity in skin tissue， along with decreased serum levels of NE， MPO， TNF-α， and VCAM-1 （P＜0.05）. In LXTZF group and RAF inhibition groups， reduced mRNA expression of RAF， MEK， ERK1， and ERK2 was observed in rat skin tissue， alongside decreased RAF protein levels and reduced p-MEK/MEK and p-ERK/ERK ratios （P＜0.05）. Compared with LXTZF + RAF agonist group， the compound glycyrrhizin group， LXTZF group， and RAF inhibitior group exhibited reduced IgA fluorescence intensity in skin tissue， decreased serum NE， MPO， TNF-α， and VCAM-1 levels， and decreased MEK mRNA expression and p-MEK/MEK ratio （P＜0.05）. ConclusionThe potential mechanism by which LXTZF treats Henoch-Schönlein purpura with blood heat syndrome may involve blocking the RAF/MEK/ERK signaling pathway in skin tissue， and suppressing excessive formation of NETs， thereby reducing IgA deposition in dermal microvessels and attenuating systemic inflammatory responses.

Objective To make transfusion management strategies for patients with history of blood transfusion and/or pregnancy by following up a patient with delayed hemolytic transfusion reactions(DHTR)caused by unexpected antibody produced after blood transfusion.Methods ABO,Rh,MN and Kidd blood group test,direct antiglobulin test,unexpected antibody screening,antibody identification,antibody titer detection,and cross-matching test were performed on a patient with DHTR.Meanwhile,suitable red blood cells were screened for subsequent treatment.Results The patient′s blood group was B,RhD(+)and CCDee,the antibody screening test and cross-matching test were negative before the first trans-fusion.After eight days,hemoglobin of the patient decreased to 57 g/L and the laboratory results indicated delayed hemoly-sis,the antibody screening was positive,and the antibody identification result was anti-M,as RBCs of the patient received typed as M+N+.After the patient received M antigen negative RBCs,the laboratory test results still indicated delayed sero-logic transfusion reaction.A new antibody arose and was identified as anti-Jka while RBCs transfused were M-N+and Jk(a+b-).Afterwards,it was effective for the patient to receive B,RhD(+),M-N+and Jk(a-b+)RBCs.Conclusion Most of the homologous antibodies produced by patients after blood transfusion will disappear within a few years.When patients undergo another transfusion,DHTR may occur because of anamnestic reaction.Establishing a transfusion management docu-ment and creating a card for patients who have already produced RBC alloantibodies can greatly reduce the occurrence of DHTR by informing doctors and staff when the next transfusion is needed.

Objective To discuss the perioperative blood transfusion strategy of patients with hereditary coagulation fac-tor deficiency through the perioperative blood coagulation management of a patient with coagulation factor Ⅺ deficiency and hemangioma of knee joint.Methods Coagulation factor activity test and gene test were used to confirm the patients with co-agulation factor Ⅺ antecedent deficiency.Perioperative bleeding risk assessment,thromboela-stogram(TEG)and coagula-tion factors detection were performed to detect the coagulation status of the patients.400 mL fresh frozen plasma was trans-fused before surgery and first day after surgery,200 mL daily for 3 days thereafter.Results The level of APTT was con-trolled from 97.1 s before operation to 36.9 s after operation.There was no obvious bleeding during operation,and the wound healed well after operation.Conclusion In the situation of lack of commercial coagulation factors,transfusion of blood products such as fresh frozen plasma can significantly improve the coagulation function and reduce the risk of intraoperative bleeding in patients with hereditary coagulation factor deficiency during perioperative period.

To explore the method of constructing the first-class undergraduate course of Treatise on Febrile Diseases. The undergraduates majoring in traditional Chinese medicine were selected as the teaching subjects. Three combinations (combination of theory and clinics, combination of science and education, and combination of ideological/political education and professional education) were adopted as the starting point. A variety of teaching methods were used. The combination of in-class and after-class and the combination of theory and clinics were implemented. Course construction was carried out from the aspects of course objectives, course resource construction, course content, organization and implementation, and course evaluation methods. Emphasis was placed on the process evaluation and formative evaluation. Questionnaire survey and final examination were used to evaluate the teaching effect. The results showed that a total of 10 first and second prizes were awarded in the provincial and municipal innovation competitions related to the theory of cold disease in the past three years. During the same time, more than 70 medical records were collected and collated, 82 research papers and 155 learning experiences were documented, five communities were visited along with the affiliated hospital and the school hospital for providing healthcare services in rural areas, and more than 20 traditional Chinese diagnosis and treatment activities were performed in these communities. The average score of students in overall course evaluation was 87. More than 94.00% (141/150) students highly evaluated the course construction. They believed that the course construction was very helpful to the theoretical study and clinical application of Treatise on Febrile Diseases, and improved their levels and abilities in classic reading, medical case evaluation and analysis, and syndrome differentiation and treatment. Students' recognition of this course has been improved and phased results have been achieved.

Objective:To investigate the feasibility of simultaneous arteriovenous enhancement of neck CT with two-stage injection of contrast agent and its effect on image quality and radiation dose.Methods:A total of 30 patients undergoing neck CT enhancement scan due to space-occupying lesions in Beijing Tongren Hospital, Capital Medical University from February to April 2022 were prospectively included as the experimental group. The neck CT enhancement scan was performed with two-stage injection of contrast agent and arteriovenous simultaneous enhancement. The dosage of contrast agent was calculated according to the patient′s body weight, and the method of two-stage injection was adopted. The dosage of contrast agent in the first stage was 0.7 ml/kg, with normal saline in the middle stage, and the second stage (began at 35 s) was 0.3 ml/kg. A total of 30 patients with gender and age matching with the experimental group from December 2021 to January 2022 were retrospectively collected as the control group. The control group was treated with the traditional arterial phase and venous phase scanning method with the dosage of 1.0 ml/kg contrast agent. The arterial phase was scanned at the 30 s and the venous phase was scanned at the 60 s. The CT values of bilateral carotid arteries and jugular veins in the experimental group were measured, the CT values of bilateral carotid arteries in the arterial phase were measured in the control group, and the CT values of bilateral carotid arteries and jugular veins in the venous phase were measured. Carotid artery enhancement score was performed for images of experimental group and control group in arterial and venous phase, and jugular vein and lesion enhancement score was performed for images of experimental group and control group in venous phase. The effective dose was calculated for both groups. The difference of carotid artery CT values between images was compared by one-way analysis of variance, and LSD method was used for pairwise comparison. The CT values of jugular vein were compared using independent sample t test. Kruskal-Wallis test was used to compare carotid artery enhancement scores, and Nemenyi method was used for pairwise comparison. Jugular vein and lesion enhancement scores and effective dose were compared by Mann-Whitney U test. Results:The CT value of carotid artery of experimental group [left (276±24) HU, right (273±25) HU] was lower than that of control group in arterial phase [left (329±33) HU, right (327±32) HU], and higher than that in the venous phase [left (147±15) HU, right (148±16) HU]. All the differences were statistically significant ( P<0.001). The CT value of jugular vein of experimental group [left (206±18) HU, right (203±19)] was higher than that of control group in the venous phase [left (154±15) HU, right (151±15)], the difference was statistically significant ( t=11.88, 11.76, both P<0.001). There was no significant difference in carotid artery enhancement score between experimental group and control group in arterial phase ( P=0.624), but the carotid artery enhancement score of the experimental group was higher than that of the control group in the venous phase, and the difference was statistically significant ( P<0.001). The scores of jugular vein and lesion enhancement in experimental group were higher than those of control group in venous phase, and the difference was statistically significant ( Z=5.01, P<0.001). The effective dose of the experimental group [2.41(2.04, 2.72) mSv] was decreased by 52.2% compared with the control group [5.04(4.18, 5.44) mSv], and the difference was statistically significant ( Z=-6.24, P<0.001). Conclusions:The neck CT enhanced scan with two-stage injection of contrast agent and arteriovenous simultaneous enhancement method can obtain comprehensive images of arterial and venous phases, and realize simultaneous enhancement of carotid artery, jugular vein and lesions, and reduce radiation dose.

【Objective】 To design and simulate routine serological experiments in transfusion techniques using RhD blood group, so as to solve the problem of difficult to obtain positive specimens in experimental teaching. 【Methods】 RhD positive red blood cells, RhD negative red blood cells and anti-D reagent were used to design and simulate the enzyme treatment experiment, absorption and elution test, antibody identification experiment and cross matching experiment of polybrene technology in transfusion techniques. 【Results】 Papain treatment of red blood cells made the agglutination of RhD positive red blood cells and IgG anti-D visible. Absorption and elution test were successfully simulated with RhD positive red blood cells and IgG anti-D reagent. The antibody identification of anti-Jk^a and anti-Fy^a was successfully simulated by creating different identification panel and panel cells made by RhD positive cells and negative cells. Cross matching test of polybrene method can also be simulated using RhD negative and positive red blood cells and IgG anti-D reagent. 【Conclusion】 RhD blood group can be used to simulate most of the routine serological experiments of blood transfusion, which can be used in the serological laboratory teaching of transfusion.

Idiopathic pulmonary fibrosis (IPF) is a progressive disease with unknown etiology and limited therapeutic options. Activation of fibroblasts is a prominent feature of pulmonary fibrosis. Here we report that lncRNA DACH1 (dachshund homolog 1) is downregulated in the lungs of IPF patients and in an experimental mouse model of lung fibrosis. LncDACH1 knockout mice develop spontaneous pulmonary fibrosis, whereas overexpression of LncDACH1 attenuated TGF-β1-induced aberrant activation, collagen deposition and differentiation of mouse lung fibroblasts. Similarly, forced expression of LncDACH1 not only prevented bleomycin (BLM)-induced lung fibrosis, but also reversed established lung fibrosis in a BLM model. Mechanistically, LncDACH1 binding to the serine/arginine-rich splicing factor 1 (SRSF1) protein decreases its activity and inhibits the accumulation of Ctnnb1. Enhanced expression of SRSF1 blocked the anti-fibrotic effect of LncDACH1 in lung fibroblasts. Furthermore, loss of LncDACH1 promoted proliferation, differentiation, and extracellular matrix (ECM) deposition in mouse lung fibroblasts, whereas such effects were abolished by silencing of Ctnnb1. In addition, a conserved fragment of LncDACH1 alleviated hyperproliferation, ECM deposition and differentiation of MRC-5 cells driven by TGF-β1. Collectively, LncDACH1 inhibits lung fibrosis by interacting with SRSF1 to suppress CTNNB1 accumulation, suggesting that LncDACH1 might be a potential therapeutic target for pulmonary fibrosis.

Surgery is the mainstay of treatment for early lung cancers, but there is still a risk of recurrence and metastasis after surgery. With the advancement of molecular biology and detection methods, detecting the level of postoperative peripheral blood minimal residual disease (MRD) in patients can dynamically monitor recurrence and determine prognosis. Due to the wide variety of MRD detection methods, uneven detection power, lack of uniform standards and prospective study validation, clinical application is still controversial. The further development of MRD detection for early stage cell lung cancer still needs technical progress, standardized detection criteria and credible clinical data.