Objective: To constructCSF1R+/-mice and to analyze their genotypes, so as to provide animal model basis for disease pathological mechanism and drug target. Methods : A linearized targeting vector was designed according to Cre/Loxp system. A Loxp site was inserted upstream of the 5th exon of theCSF1Rgene, and a neomycin resistance box with Loxp sites on both sides was inserted downstream of the 5th exon. The linearized targeting vector was electroporated into embryonic stem cells. The correctly targeted embryonic stem cells were injected into the blastocysts of C57BL/6J mice to obtain chimeric mice, which were bred with Zp3-Cre mice. The newborn mice were numbered 9-14 days after birth and their tails were cut. The DNA of the mice was extracted, and the genotype of the mice was identified by polymerase chain reaction and agarose gel electrophoresis. The expression of CSF1R in mouse macrophages was detected by flow cytometry. The expression of CSF1R in mouse tissues was detected by Western blot. Results: The results of agarose gel electrophoresis showed that 453 bp bands were amplified in wild type mice, and 453 bp and 650 bp bands were amplified in heterozygous mice. The results of flow cytometry showed that the expression of CSF1R in peritoneal macrophages and bone marrow-derived macrophages of CSF1R heterozygous mice was lower than that of WT group(P<0.05). The results of Western blot showed that the expression of CSF1R in spleen, kidney and brain tissue of CSF1R heterozygous group was lower than that of WT group(P<0.05). Conclusion CSF1R+/-mice are successfully constructed, reproduced and identified, which provides an animal model basis for further revealing the potential mechanism of CSF1R in immune regulation.

Objective : To construct triggering receptor expressed on myeloid cells 2(TREM2) gene knockout(TREM2-/-) mice using CRISPR/Cas9 technology, to breed TREM2-/- mice and to analyze the genotype of TREM2-/- mice. Methods : CRISPR/Cas9 technology was used to selectively knock out exon 2-3 regions of TREM2 gene to construct a TREM2-/- mouse model, and the genetic background of all mice was C57BL/6J. Polymerase chain reaction(PCR) was used to identify the genotype of mice. Quantitative real-time PCR(qPCR) and Western blot were used to detect the expression level of TREM2 in major tissues of mice, and the authenticity and scientific nature of PCR identification results were verified from mRNA level and protein level. According to the sgRNA sequence, the possible off-target sites were predicted on the CCTop website, and the tail DNA of mice was extracted and amplified by PCR and then Sanger sequencing was performed to detect whether there was off-target effect in TREM2-/- mice. Results : TREM2-/- mice were successfully constructed by CRISPR/Cas9 technology, and the mice were genotyped. The results of agarose gel electrophoresis showed that the mouse genotype with only 415 bp band amplified was wild type(WT), the mouse genotype of the 449 bp band amplified only was TREM2-/-, and the mouse genotypes amplified with 415 bp and 449 bp double bands were heterozygous. qPCR results showed that compared with WT mice, the mRNA expression of TREM2 in heart and brain tissues of TREM2-/- mice was down-regulated(P-/- mice was reduced(P-/- mice. Conclusion TREM2-/- mice are successfully constructed and bred, a reliable genotype identification method is established, the genetic stability of the mouse model is verified, which will provide an important genetic animal model for the study of TREM2 gene function.

Objective To retrieve the anticoagulation management related evidences in postoperative patients with heart valve operation to provide an evidence-based basis for medical professionals conducting the antithrombotic therapy in postoperative patients with heart valve operation.Methods The related evidences of anticoagulation management after heart valve surgery were retrieved from major databases and websites at home and abroad based on the"6S"evidence resource pyramid modeling system.The retrieval time was from January 1,2017 to February 29,2024.Two researchers trained by evidence-based care evaluated the quality of the literatures and extracted the relevant evidences meeting the criteria by combining with clinical situation.Results A total of 12 literatures were included,including 5 guidelines,4 clinical practices and 3 expert consen-sus.Thirty pieces of best evidences were summarized in eight aspects:contraindications,postoperative an-tithrombotic regimens,warfarin use method,vitamin K use precautions,daily life precautions,anticoagulation complications,long term perioperative anticoagulation bridging and complicating pregnancy.Conclusion The evidences summarized in this study could provide a reference for standardizing anticoagulation management in postoperative pa-tients with heart valve operation and for clinical healthcare workers in order to standardize the anticoagulation therapy after heart valve replacement and reduce the occurrence of anticoagulation-related complications.

Objective:To construct an integrated early rehabilitation program for healthcare and rehabilitation system based on the information-motivation-behavioral skills (IMB) model and evaluate its application effect in patients treated with total hip arthroplasty (THA).Methods:Construction of the rehabilitation program: An integrated research team was established, composed of head burse of orthopedics, orthopedic nursing specialists, orthopedic surgeons, anesthesiologists, and rehabilitation therapists. Considering the key points of perioperative early rehabilitation of THA patients, an integrated early rehabilitation program was constructed based on the IMB model through literature review and expert panel method. Clinical application of the rehabilitation program: A retrospective cohort study was conducted to analyze the clinical data of 100 THA patients admitted to Qingdao Municipal Hospital from March to December 2023, including 47 males and 53 females, aged 60-85 years [(69.8±5.5)years]. Patients were divided into two groups according to their admission time: 50 patients admitted from March to July 2023, receiving routine care (routine care group) and 50 admitted from August to December 2023, receiving intervention through an early rehabilitation program of an integrated healthcare and rehabilitation system based on routine care (integrated care group). The first postoperative ambulation time and length of hospital stay were compared between the two groups. The Harris hip function score was used to assess hip function in both groups at 3 days, 1, 3, and 6 months postoperatively; the Barthel index was used to assess the daily living self-care capacity in both groups preoperatively, at 1, 3, and 6 months postoperatively. The incidence of complications within 6 months after surgery was compared between the two groups.Results:An early integrated rehabilitation program based on the IMB model comprised 25 specific measures of three domains: information support, motivational intervention, and behavioral skills. All the patients were followed up for 6 months. The first postoperative ambulation time and length of hospital stay in the integrated care group were (1.3±0.5)days and (7.4±2.3)days, shorter than (1.5±0.5)days and (8.5±2.3)days in the routine care group ( P<0.05). There were no statistically significant differences in the Harris hip function scores at 3 days postoperatively or preoperative Barthel index between the two groups ( P>0.05). At 1, 3, and 6 months postoperatively, the Harris hip function scores in the integrated care group were (80.3±6.0)points, (88.6±5.2)points, and (92.5±4.1)points, respectively, higher than (75.1±6.3)points, (84.2±5.7)points, and (88.0±5.2)points in the routine care group ( P<0.01); the Barthel index in the integrated care group were (79.2±8.7)points, (87.7±5.7)points, and (92.3±4.9)points, respectively, higher than (72.1±9.0)points, (83.5±6.6)points, and (88.6±5.0)points in the routine care group ( P<0.01). At 6 months postoperatively, the incidence of complications in the integrated care group was 4% (2/50), lower than 16% (8/50) in the routine care group ( P<0.05). Conclusion:Compared with the routine care, an integrated early rehabilitation program for medical care and health based on the information-motivation-behavioral skills model for THA patients can shorten the first postoperative ambulation time and length of hospital stay, restore hip joint function, improve daily living self-care capacity and reduce the incidence of complications.

This article reports the diagnosis and treatment of a patient with diffuse large B-cell lymphoma (DLBCL) initially manifested as acquired von Willebrand syndrome (AvWS), along with a literature review. The patient, a 22-year-old male, was diagnosed with hereditary von Willebrand disease at the initial stage due to repeated mucosal bleeding, and was later diagnosed with DLBCL (non germinal center type, Ann Arbor stage Ⅳ) due to chest wall mass. Through chemotherapy combined with autologous hematopoietic stem cell transplantation and zebutinib maintenance therapy, the patient achieved sustained complete remission, and the coagulation function returned to normal. This case provides an important reference for the diagnosis and treatment of lymphoma associated AvWS, and highlights the importance of early recognition of basic diseases.

There is a large gap between production and demand of plant oil in China, which leads to the heavy reliance on imports. Diacylglycerol acyltransferase (DGAT) and phospholipid: diacylglycerol acyltransferase (PDAT) are two key enzymes responsible for the synthesis of triacylglycerol, thereby affecting the yield and quality of plant oil. This paper comprehensively reviews the research progress in DGAT and PDAT in terms of their biological functions in plant oil synthesis, the molecular mechanisms of regulating plant lipid metabolism, growth, and development under stress, and their roles in driving oil synthesis under the background of synthetic biology. Furthermore, future research and application of DGAT and PDAT are prospected. This review aims to provide a basis for deeply understanding the molecular mechanism of plant oil synthesis and improving the quality and productivity of oil crops by the utilization of DGAT and PDAT genes.
Diacylglycerol O-Acyltransferase/physiology* ; Plant Oils/metabolism* ; Acyltransferases/metabolism* ; Lipid Metabolism/genetics* ; Gene Expression Regulation, Plant ; Triglycerides/biosynthesis*

Objective To explore the mechanism of malt alcohol extract improving depression-like behavior induced by CUMS in rats by regulating gut microbiota.Methods The depression model of rats was established using an 8-weeks CUMS procedure,and the administration group was given low(59.6 mg·kg-1)and high(178.8 mg·kg-1)doses of malt alcohol extract,respectively.The depression-like behavior of rats was evaluated by classic behavioral test.The composition of intestinal microbiota of rats was analyzed by 16S rRNA sequencing.The morphological changes of colon were observed by hematoxylin and eosin(HE),the expression of ZO-1 and Occludin in colon was detected by immunofluorescence(IF),and the expression of IL-10,IL-1βand 5-HT were detected by ELISA.Results The low dose of malt alcohol extract attenuated the depressive behavior and restored the expression of 5-HT in the brain of CUMS rats.16S rRNA sequencing results showed that the diversity and relative abundance of gut microbiota changed after treatment with the low dose of malt alcohol extract.ELISA results showed that the low dose of malt alcohol extract significantly reversed the CUMS-induced reduction of IL-10 and elevation of IL-1 β.HE results showed that the low dose of malt alcohol extract significantly ameliorated CUMS-induced structural damage in colon.IF results showed increased protain expression of intestinal epithelial barrier tight junction proteins ZO-1 and Occludin by the low dose of malt alcohol extract.Conclusion The low dose of malt alcohol extract can ameliorate CUMS-induced depressive-like behavior in rats by modulating intestinal flora,restoring 5-HT expression in the brain,inhibiting inflammation,and repairing the intestinal barrier.

OBJECTIVE To investigate the protective effect and mechanism of saikosaponin C(SSC)on acute liver injury(ALI)in mice induced by carbon tetrachloride(CCl4)based on serum metabolomics.METHODS Forty mice were divided into blank group(water),model group(water),positive control drug group(Biphenyl diester drop pills,150 mg/kg),and SSC low-and high-dose groups(2.5,10 mg/kg)using the random number table method,with 8 mice in each group.They were given water/relevant drugs,once a day,for 7 consecutive days.One hour after the last administration,all mice were intraperitoneally injected with 0.2％CCl4 olive oil to induce ALI model,except for the blank group.After 17 hours of the modeling,the liver index of mice was calculated.The levels of aspartate aminotransferase(AST),alanine aminotransferase(ALT),lactate dehydrogenase(LDH),tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),and IL-1β in serum of mice were detected.The histopathological changes of liver tissue were observed.Meanwhile,the serum metabolomics of mice were analyzed by liquid chromatography-mass spectrometry.RESULTS Compared with the blank group,the levels of liver index,ALT,AST,LDH,TNF-α,IL-6,and IL-1β in the model group were significantly increased(P＜0.01).Hepatocytes were edema,vacuolar degeneration,more necrosis,and a large number of inflammatory cells were infiltrated.Compared with the model group,liver index and serum index levels of mice were significantly decreased(P＜0.05 or P＜0.01),accompanied by marked improvement in histopathological damage to the liver tissue.The metabolomics results showed that compared with the model group,there were 63 up-regulated and 256 down-regulated differential metabolites in the serum of mice in the SSC high-dose group,including prostaglandin B2,20-hydroxy-leukotriene B4,5-hydroxy-L-tryptophan,7α-hydroxycholesterol,etc.;these metabolites were primarily involved in metabolic pathways such as arachidonic acid metabolism,5-hydroxytryptamine synapse,primary bile acid biosynthesis.CONCLUSIONS SSC exerts a protective effect against CCl4-induced ALI by down-regulating the level of key metabolites such as prostaglandin B2 and 20-hydroxy-leukotriene B4,and then ruducing metabolic pathways such as arachidonic acid metabolism,5-hydroxytryptamine synapse,and primary bile acid biosynthesis.

Objective To explore the mechanism of malt alcohol extract improving depression-like behavior induced by CUMS in rats by regulating gut microbiota.Methods The depression model of rats was established using an 8-weeks CUMS procedure,and the administration group was given low(59.6 mg·kg-1)and high(178.8 mg·kg-1)doses of malt alcohol extract,respectively.The depression-like behavior of rats was evaluated by classic behavioral test.The composition of intestinal microbiota of rats was analyzed by 16S rRNA sequencing.The morphological changes of colon were observed by hematoxylin and eosin(HE),the expression of ZO-1 and Occludin in colon was detected by immunofluorescence(IF),and the expression of IL-10,IL-1βand 5-HT were detected by ELISA.Results The low dose of malt alcohol extract attenuated the depressive behavior and restored the expression of 5-HT in the brain of CUMS rats.16S rRNA sequencing results showed that the diversity and relative abundance of gut microbiota changed after treatment with the low dose of malt alcohol extract.ELISA results showed that the low dose of malt alcohol extract significantly reversed the CUMS-induced reduction of IL-10 and elevation of IL-1 β.HE results showed that the low dose of malt alcohol extract significantly ameliorated CUMS-induced structural damage in colon.IF results showed increased protain expression of intestinal epithelial barrier tight junction proteins ZO-1 and Occludin by the low dose of malt alcohol extract.Conclusion The low dose of malt alcohol extract can ameliorate CUMS-induced depressive-like behavior in rats by modulating intestinal flora,restoring 5-HT expression in the brain,inhibiting inflammation,and repairing the intestinal barrier.

Objective:To construct an integrated early rehabilitation program for healthcare and rehabilitation system based on the information-motivation-behavioral skills (IMB) model and evaluate its application effect in patients treated with total hip arthroplasty (THA).Methods:Construction of the rehabilitation program: An integrated research team was established, composed of head burse of orthopedics, orthopedic nursing specialists, orthopedic surgeons, anesthesiologists, and rehabilitation therapists. Considering the key points of perioperative early rehabilitation of THA patients, an integrated early rehabilitation program was constructed based on the IMB model through literature review and expert panel method. Clinical application of the rehabilitation program: A retrospective cohort study was conducted to analyze the clinical data of 100 THA patients admitted to Qingdao Municipal Hospital from March to December 2023, including 47 males and 53 females, aged 60-85 years [(69.8±5.5)years]. Patients were divided into two groups according to their admission time: 50 patients admitted from March to July 2023, receiving routine care (routine care group) and 50 admitted from August to December 2023, receiving intervention through an early rehabilitation program of an integrated healthcare and rehabilitation system based on routine care (integrated care group). The first postoperative ambulation time and length of hospital stay were compared between the two groups. The Harris hip function score was used to assess hip function in both groups at 3 days, 1, 3, and 6 months postoperatively; the Barthel index was used to assess the daily living self-care capacity in both groups preoperatively, at 1, 3, and 6 months postoperatively. The incidence of complications within 6 months after surgery was compared between the two groups.Results:An early integrated rehabilitation program based on the IMB model comprised 25 specific measures of three domains: information support, motivational intervention, and behavioral skills. All the patients were followed up for 6 months. The first postoperative ambulation time and length of hospital stay in the integrated care group were (1.3±0.5)days and (7.4±2.3)days, shorter than (1.5±0.5)days and (8.5±2.3)days in the routine care group ( P<0.05). There were no statistically significant differences in the Harris hip function scores at 3 days postoperatively or preoperative Barthel index between the two groups ( P>0.05). At 1, 3, and 6 months postoperatively, the Harris hip function scores in the integrated care group were (80.3±6.0)points, (88.6±5.2)points, and (92.5±4.1)points, respectively, higher than (75.1±6.3)points, (84.2±5.7)points, and (88.0±5.2)points in the routine care group ( P<0.01); the Barthel index in the integrated care group were (79.2±8.7)points, (87.7±5.7)points, and (92.3±4.9)points, respectively, higher than (72.1±9.0)points, (83.5±6.6)points, and (88.6±5.0)points in the routine care group ( P<0.01). At 6 months postoperatively, the incidence of complications in the integrated care group was 4% (2/50), lower than 16% (8/50) in the routine care group ( P<0.05). Conclusion:Compared with the routine care, an integrated early rehabilitation program for medical care and health based on the information-motivation-behavioral skills model for THA patients can shorten the first postoperative ambulation time and length of hospital stay, restore hip joint function, improve daily living self-care capacity and reduce the incidence of complications.