1.GRK2 inhibits Flt-1+ macrophage infiltration and its proangiogenic properties in rheumatoid arthritis.
Xuezhi YANG ; Yingjie ZHAO ; Qi WEI ; Xuemin ZHU ; Luping WANG ; Wankang ZHANG ; Xiaoyi LIU ; Jiajie KUAI ; Fengling WANG ; Wei WEI
Acta Pharmaceutica Sinica B 2024;14(1):241-255
Rheumatoid arthritis (RA) is an autoimmune disease with a complex etiology. Monocyte-derived macrophages (MDMs) infiltration are associated with RA severity. We have reported the deletion of G-protein-coupled receptor kinase 2 (GRK2) reprograms macrophages toward an anti-inflammatory phenotype by recovering G-protein-coupled receptor signaling. However, as more GRK2-interacting proteins were discovered, the GRK2 interactome mechanisms in RA have been understudied. Thus, in the collagen-induced arthritis mouse model, we performed genetic GRK2 deletion using GRK2f/fLyz2-Cre+/- mice. Synovial inflammation and M1 polarization were improved in GRK2f/fLyz2-Cre+/- mice. Supporting experiments with RNA-seq and dual-luciferase reporter assays identified peroxisome proliferator-activated receptor γ (PPARγ) as a new GRK2-interacting protein. We further confirmed that fms-related tyrosine kinase 1 (Flt-1), which promoted macrophage migration to induce angiogenesis, was inhibited by GRK2-PPARγ signaling. Mechanistically, excess GRK2 membrane recruitment in CIA MDMs reduced the activation of PPARγ ligand-binding domain and enhanced Flt-1 transcription. Furthermore, the treatment of mice with GRK2 activity inhibitor resulted in significantly diminished CIA pathology, Flt-1+ macrophages induced-synovial inflammation, and angiogenesis. Altogether, we anticipate to facilitate the elucidation of previously unappreciated details of GRK2-specific intracellular signaling. Targeting GRK2 activity is a viable strategy to inhibit MDMs infiltration, affording a distinct way to control joint inflammation and angiogenesis of RA.
2.Preliminary Construction of the "Eye-Brain" Sleep Regulation Theory Based on The Inner Canon of YellowEmperor (《黄帝内经》) and Modern Medicine
Lihong LI ; Fengling LIANG ; Zhaoshun LYU ; Wei JIN ; Conghua JI
Journal of Traditional Chinese Medicine 2024;65(20):2092-2096
We analysed the similarities between theory of The Inner Canon of Yellow Emperor (《黄帝内经》) and modern sleep medicine on sleep and wakefulness, and initially constructed the "eye-brain" sleep regulation system. The similarities between the sleep theories in The Inner Canon of Yellow Emperor and modern sleep medicine research are described from three perspectives: the anatomical basis of sleep physiology, the pathogenesis of sleep disorder pathology, and the mechanism of sleep disorder treatment. It is found that the regulation of sleep and wakefulness in both medicine theories is based on the "eye" and "brain" as the main control centres, and there exists a "eye-brain" pathway to regulate sleep and wakefulness in the human body. Based on this, the "eye-brain" sleep regulation theory is constructed, in order to find an effective way to solve the pathogenesis and treatment strategy of sleep disorders.
3.Research progress on rehabilitation motivation assessment tools for stroke patients
Tao XIONG ; Xuemei TAN ; Jing LUO ; Yang LI ; Yuxi ZHENG ; Fengling LI ; Xuemei WEI ; Lijun CUI ; Lanjun LUO
Chinese Journal of Nursing 2024;59(7):890-896
The rehabilitation compliance of stroke patients is generally low.Evaluating the rehabilitation motivation of patients is helpful to promote the rehabilitation management of patients,enhance the rehabilitation enthusiasm and compliance of patients,and improve the rehabilitation outcome.This paper reviews the existing stroke patients rehabilitation motivation assessment tools,and expounds the main contents,application status,characteristics and limitations of stroke patients rehabilitation motivation assessment tools,in order to provide references for the appropriate selection of clinical assessment tools,the rehabilitation management of stroke patients and the development of domestic localized stroke rehabilitation motivation assessment tools.
4.Eukaryotic expression,purification and activity identification of human His-GRK2 recombinant protein
Liping Jiang ; Luying Chen ; Jiajie Kuai ; Fengling Wang ; Hao Li ; Yanling Guan ; Yang Ma ; Chenchen Han ; Wei Wei
Acta Universitatis Medicinalis Anhui 2023;58(2):179-184
Objective:
To construct a human G protein-coupled receptor kinase 2 ( GRK2) eukaryotic expression system.
Methods:
The primers were designed ,and the His-GRK2 target gene was amplified by PCR using the Pans-EGFP-GrK2 (full-length) gene as the template.The His-GRK2 target gene was connected to the pcDNA3.1EGFP eukaryotic expression vector. The pcDNA3. 1-EGFP-His-GRK2 plasmid was transfected into HEK 293T cells.48 h later,the expression of GRK2 protein was detected by Western blot,and the GRK2 protein was purified by nickel chelated magnetic bead method.The purification of GRK2 protein was detected by Coomassie bright blue staining and Western blot,and the activity of GRK2 protein was detected by His pull down.
Results :
The results of double enzyme digestion and sequencing showed that pcDNA3. 1-EGFP-His-GRK2 eukaryotic expression plasmid was successfully constructed.Western blot analysis showed that the molecular weight of GRK2 protein was about 80 ku,indicating that GRK2 protein was successfully expressed in HEK 293T cells (t = 6. 433,P = 0. 003) .GRK2 protein was purified by nickel chelated magnetic beads.His pull down experiment results showed that GRK2 was bound to prostaglandin E2 receptor 4 (EP4) ,suggesting that GRK2 protein had biological activity (t = 13. 5,P = 0. 000 2) .
Conclusion
The pcDNA3.1-EGFP-His-GRK2 eukaryotic expression plasmid was correctly sequenced and the GRK2 recombinant plasmid was successfully constructed.The GRK2 recombinant plasmid was successfully expressed in eukaryotic cells HEK 293T and the protein expressed was biologically active.
5.Explanation of Diagnosis for occupational radiation thyroid disease
Wei GUO ; Fengling ZHAO ; Quanfu SUN ; Yumin LYU
Chinese Journal of Radiological Medicine and Protection 2023;43(12):1022-1026
In order to accurately master and understand the national occupational health standard Diagnosis for Occupational Radiation Thyroid Disease(GBZ 101-2020), the significance of the publication of this standard, the relevant background, the basic principles of the revision, the main basis for technical content revision, and the application of the standard are described. To provide guidance for t the physcians for occupational disease diagnosiss to understand the diagnostic principles, make correct diagnosis and avoid confusion in using the standard, so as to better protect the occupational health rights of radiation workers.
6.Review of lactation management in women following perinatal loss
Fengling LI ; Zhaoxia CHEN ; DUAN S DIANA ; Xiuhua ZHANG ; Lijun CUI ; Xuemei WEI ; Hongbo QI ; Qi SHI
Chinese Journal of Nursing 2023;58(22):2806-2811
Perinatal loss is one of the most serious childbirth trauma,which seriously affects maternal physical and mental health.At present,the management strategy of maternal lactation can be divided into two aspects:milk return and milk donation.Breast milk donation may build new values of perinatal loss,which can promote maternal physical and mental recovery.This article reviewed the conception,status,motivations,facilitators,barriers and interventions of lactation management in women following perinatal loss,so as to provide bases for improving the awareness of obstetric nursing staff,quality of obstetric care and future research on lactation management in women following perinatal loss in China.
7.Epidemiological investigation of SARS-CoV-2 infection in maintenance hemodialysis patients in Jiangsu province during the outbreak of SARS-CoV-2
Guang YANG ; Yifei GE ; Yaoyu HUANG ; Jizhuang LOU ; Chunming JIANG ; Guoyuan LU ; Fengling CHEN ; Jiansong SHEN ; Xiaolan CHEN ; Houyong DAI ; Changhua LIU ; Min YANG ; Xiurong LI ; Zhuxing SUN ; Liang WANG ; Bin LIU ; Donghui ZHENG ; Yong XU ; Maojie CHEN ; Ling WANG ; Yilai ZHANG ; Xu ZHANG ; Jianqiang HE ; Liyuan ZHANG ; Huiting WAN ; Honglei GUO ; Jiahui YANG ; Wei XU ; Changying XING ; Huijuan MAO
Chinese Journal of Nephrology 2023;39(12):895-902
Objective:To investigate the epidemiology of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection in patients with maintenance hemodialysis (MHD) in Jiangsu province during SARS-CoV-2 pandemic in China from December 7, 2022 to January 27, 2023, and to analyze the influencing factors of all-cause death.Methods:It was a multi-center cross-sectional investigation. Structured questionnaire was used to collect patient information by medical staff of each hemodialysis center (room) as investigators. Part of the demography data and laboratory examination data came from the Jiangsu Province Hemodialysis Data Information System. MHD patients from hemodialysis centers (rooms) at all levels of medical institutions and independent hemodialysis institutions in Jiangsu province during the outbreak of SARS-CoV-2 infection were included, and the clinical characteristics and all-cause mortality of confirmed and suspected cases of SARS-CoV-2 infection were analyzed.Results:Questionnaire surveys and data analysis on 57 278 patients in 407 hemodialysis centers (rooms) were completed, accounting for 90.41% of the total number of MHD patients (63 357 cases) in Jiangsu province during the same period. There were 24 038 cases (41.97%) of SARS-CoV-2 infection and 14 805 cases (25.85%) of suspected infection, which were widely distributed in all dialysis centers in Jiangsu province. After clinical classification of 38 843 confirmed and suspected SARS-CoV-2 infection cases, 3 662 cases were severe and critical cases, accounting for 9.43% of the infected and suspected cases. Among the patients who had completed the questionnaires, there were 1 812 all-cause deaths, with an all-cause mortality rate of 3.16%. Multivariate logistic regression analysis showed that elderly (taking ≤50 years as a reference, 51-59 years: OR=1.583, 95% CI 1.279-1.933, P=0.001; 60-69 years: OR=3.972, 95% CI 3.271-4.858, P<0.001; 70-79 years: OR=7.236, 95% CI 5.917-8.698, P<0.001; ≥80 years: OR=11.738, 95% CI 9.459-14.663, P<0.001), male ( OR=1.371, 95% CI 1.229-1.529, P<0.001), and co-infection with hepatitis B virus (HBV) (positive serum HBV surface antigen, OR=0.629, 95% CI 0.484-0.817, P<0.001) were independent influencing factors for all cause mortality. Receiver-operating characteristic curve analysis showed that the area under the curve for male, age and current HBV infection prediction of all-cause death was 0.529 ( P<0.001), 0.724 ( P<0.001) and 0.514 ( P=0.042), respectively, and the cut-off value for age prediction of all-cause death was 65.5 years old. Compared with patients without HBV infection, MHD patients with HBV infection significantly reduced the proportion of severe and critically ill patients, all-cause hospitalizations and all cause deaths when infected with SARS-CoV-2 (4.99% vs. 6.41%, χ2=6.136, P=0.013; 8.90% vs. 11.44%, χ2=11.662, P<0.001; 2.01% vs. 3.37%, χ2=10.713, P=0.001, respectively). Conclusion:The MHD patients in Jiangsu province are susceptible to SARS-CoV-2. Elderly age and male gender are independent risk factors for death in MHD patients during the epidemic, while the HBV infection may be a protective factor for death of MHD patients infected with SARS-CoV-2.
8.Prokaryotic expression and polyclonal antibody preparation of LRRC15 protein, an excretory-secretory antigen of Cysticercus cellulosae
Qianqian WANG ; Shimin WANG ; Qianfei WANG ; Fengling YUAN ; Wei HE ; Lizhu LI ; Biying ZHOU
Chinese Journal of Endemiology 2023;42(9):704-709
Objective:To construct a recombinant plasmid pET30a-leucine-rich repeat (LRR) containing 15 (LRRC15) of Taenia solium, prokaryotically express and purify the LRRC15 recombinant protein, and prepare a rabbit polyclonal antibody. Methods:The LRRC15 protein encoding gene of Taenia solium was obtained by whole gene synthesis; it was cloned into pET30a vector, and the recombinant plasmid pET30a-LRRC15 was constructed and identified by double-enzyme PCR; the recombinant plasmid was transformed into competent cells of Escherichia coli BL21 (DE3), and the recombinant protein LRRC15 was induced to express by isopropyl-beta-D-thiogalactopyranoside (IPTG), the expression product was analyzed and identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE); the LRRC15 recombinant protein was purified by Ni-IDA affinity columns, the purified recombinant protein was analyzed and identified by SDS-PAGE, and the specificity of the purified recombinant protein was identified by Western blot (WB); the New Zealand rabbits were immunized with purified LRRC15 recombinant protein to prepare polyclonal antibodies against LRRC15, and the potency of the purified polyclonal antibody was determined by indirect enzyme-linked immunosorbent assay (ELISA). Results:After PCR identification, a band with a length of 1 506 bp was amplified, which was consistent with the LRRC15 gene; after SDS-PAGE and WB identification, the LRRC15 target protein with a relative molecular mass ( Mr) of about 55.36 × 10 3 was obtained; after immunizing New Zealand rabbits with purified LRRC15 recombinant protein, a polyclonal antibody against LRRC15 was obtained, and its potency was 1∶1 587 200. Conclusion:The recombinant plasmid pET30a-LRRC15 is successfully constructed, the LRRC15 recombinant protein of Taenia solium is prepared, and a high purity and high potency rabbit anti polyclonal antibody against LRRC15 recombinant protein is obtained.
9.Evaluation and analysis on diagnostic criteria for common occupational radiation-induced diseases among radiation workers in some provinces and cities of China
Wei GUO ; Fengling ZHAO ; Zhiwei XING ; Ling HE ; Wei LIU ; Yulong LIU ; Shouzheng WANG ; Zaiqing HE ; Wei CHEN ; Guizhi CHEN
Chinese Journal of Radiological Medicine and Protection 2023;43(8):620-626
Objective:To investigate the application of diagnostic criteria for common occupational radiation-induced diseases to radiation workers, in order to provide a basis for the revision, publicity and standardization of the standards.Methods:Radiation workers were selected from 1 city, 7 provinces and 1 corporation by using cluster random sampling method from January 2021 to May 2021. Awareness of the criteria and the effects of ionizing radiation, and the suggestions for diagnostic works were investigated and analyzed.Results:A total of 2 839 radiation workers were investigated. There were differences in the awareness of different diagnostic criteria, the inclusions in complex diagnostic criteria, the materials required for applying for diagnosis, and the ways of knowing the diagnostic criteria( χ2=416.06, 2 924.14, 83.45, 895.67, 815.94, P<0.001). The correct understanding rates of deterministic effects and stochastic effects were 80.63% and 43.64%, respectively. The acceptance rates in applicable materials were 96.79% for occupational exposure history, 94.72% for occupational health monitoring records and 93.55% for individual monitoring of occupational exposure, respectively. Pre-employment training rate was 80.20%, on-job training rate was 81.19%, and untrained rate was 3.77%. The suggestions to the diagnosis of occupational radiation-induced diseases are to strengthen training, pay attention to individual monitoring, occupational health examination, and strengthen health supervision and law enforcement. Conclusions:Radiation workers have a low awareness rate of certain diagnostic standards and a high awareness rate of diagnostic procedures. Publicity and training of health effects of ionizing radiation and diagnostic criteria of occupational radiation-induced diseases should be strengthened. Diagnostic procedure should be optimized.
10.Anatomic study and clinical application of iliac crest chimeric tissue flap.
Ziyang ZHANG ; Ling YUAN ; Wenhu JIN ; Fengling ZHANG ; Zairong WEI
Chinese Journal of Reparative and Reconstructive Surgery 2023;37(11):1418-1422
OBJECTIVE:
To conduct anatomical study on the iliac crest chimeric tissue flap and summarize its effectiveness of clinical application in repairing limb wounds.
METHODS:
Latex perfusion and anatomical study were performed on 6 fresh adult cadaver specimens with 12 sides, to observe the initial location, distribution, quantity, and direction of the common circumflexa iliac artery, the deep circumflexa iliac artery, and the superficial circumflexa iliac artery, and to measure their initial external diameter. Between December 2020 and September 2022, the iliac crest chimeric tissue flap repair was performed on 5 patients with soft tissue of limbs and bone defects. There were 3 males and 2 females, with an average age of 46 years (range, 23-60 years). Among them, there were 3 cases of radii and skin soft tissue defects and 2 cases of tibia and skin soft tissue defects. The length of bone defects was 4-8 cm and the area of skin soft tissue defects ranged from 9 cm×5 cm to 15 cm×6 cm. The length of the iliac flap was 4-8 cm and the area of skin flap ranged from 12.0 cm×5.5 cm to 16.0 cm×8.0 cm. The donor sites were directly sutured.
RESULTS:
Anatomical studies showed that there were 10 common circumflex iliac arteries in 5 specimens, which originated from the lateral or posterolateral side of the transition between the external iliac artery and the femoral artery, with a length of 1.2-1.6 cm and an initial external diameter of 0.8-1.4 mm. In 1 specimen without common circumflexa iliac artery, the superficial and deep circumflex iliac arteries originated from the external iliac artery and the femoral artery, respectively, while the rest originated from the common circumflex iliac artery. The length of superficial circumflex iliac artery was 4.6-6.7 cm, and the initial external diameter was 0.4-0.8 mm. There were 3-6 perforator vessels along the way. The length of deep circumflex iliac artery was 7.8-9.2 cm, and the initial external diameter was 0.5-0.7 mm. There were 3-5 muscular branches, 4-6 periosteal branches, and 2-3 musculocutaneous branches along the way. Based on the anatomical observation results, all iliac crest chimeric tissue flaps were successfully resected and survived after operation. The wounds at recipient and donor sites healed by first intention. All patients were followed up 8-24 months, with an average of 12 months. The tissue flap has good appearance and soft texture. X-ray film reexamination showed that all the osteotomy healed, and no obvious bone resorption was observed during follow-up.
CONCLUSION
The common circumflex iliac artery, deep circumflex iliac artery, and superficial circumflex iliac artery were anatomically constant, and it was safe and reliable to use iliac crest chimeric tissue flap in repairing the soft tissue and bone defects of limbs.
Adult
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Male
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Female
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Humans
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Middle Aged
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Plastic Surgery Procedures
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Ilium/surgery*
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Perforator Flap/blood supply*
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Skin Transplantation/methods*
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Lower Extremity/surgery*
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Soft Tissue Injuries/surgery*
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Treatment Outcome


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