G protein-coupled receptor kinase 2 (GRK2) participates in the phosphorylation and desensitization of G protein-coupled receptor (GPCR), impacting various biological processes such as inflammation and cell proliferation. Dysregulated expression and activity of GRK2 have been reported in multiple cells in rheumatoid arthritis (RA). However, whether and how GRK2 regulates synovial hyperplasia and fibroblast-like synoviocytes (FLSs) proliferation is poorly understood. In this study, we investigated the regulation of GRK2 and its biological function in RA. We found that GRK2 transmembrane activity was increased in FLSs of RA patients and collagen-induced arthritis (CIA) rats. Additionally, we noted a positive correlation between high GRK2 expression on the cell membrane and serological markers associated with RA and CIA. Immunoprecipitation-mass spectrometry and pull-down analyses revealed tumor necrosis factor receptor-associated factor 2 (TRAF2) as a novel substrate of GRK2. Furthermore, surface plasmon resonance (SPR) and molecular docking assays determined that the C-terminus of GRK2 binds to the C-terminus of TRAF2 at the Gln340 residue. GRK2 knockdown and the GRK2 inhibitor CP-25 attenuated synovial hyperplasia and FLS proliferation in CIA both in vitro and in vivo by decreasing GRK2 membrane expression and activity. Mechanistically, increased GRK2 transmembrane activity contributed to the recruitment of TRAF2 on the cell membrane, promoting GRK2-TRAF2 interactions that facilitate the recruitment of the E3 ubiquitin ligase TRIM47 to TRAF2. This enhanced TRAF2 Lys63 polyubiquitylation and induced nuclear factor (NF)-κB activation, leading to synovial hyperplasia and abnormal proliferation of FLSs. Our study provides a mechanistic and preclinical rationale for further evaluation of GRK2 as a therapeutic target for RA.

Personality development theory starts from the physiological needs of individuals,proposes the"two-part personality structure"with"subconscious"and"consciousness"as the subject,and then deduces the"three-part personality structure"with"id,ego,and superego"as the subject.Based on this framework,an instinctive personality dynamics schema was constructed to explore the causes of self-imbalance.Taking these as a foothold,the paper deduced the formation of moral injury from the personality dynamic schema,found the moral anxiety from the self-contradiction of ego,id,and superego,as well as deduced the representation form of moral injury based on Freud's"anxiety".Furthermore,the differences in individual personality development are introduced into the study of moral injury to distinguish between anxiety and guilt in the symptoms of moral injury,in order to better understand moral trauma.

Moral resilience is the ability to positively respond to moral dilemmas,consisting of firm beliefs,tenacious willpower,harmonious relationships,and a peaceful mind.The Chinese nation is a resilient nation,its excellent traditional Chinese culture is vast and profound,containing extremely rich ideas about moral resilience.For example,"being erudite and determined,eager to ask and thinking closely,and benevolence lies in it"and"if one aspires to pursue benevolence,they will not do evil things".The ancients believed that human resilience could correct moral motives and enable people to overcome difficulties and forge ahead in adversity.The intellectual wisdom of the ancients can provide advice and consultation for cultivating moral resilience today.Therefore,need drawing on traditional culture to strengthen moral resilience.

To analyze the infection of chlamydia (CT) and gonorrhea (NG) in female infertility and male infertility population, and to explore the correlation between CT and NG infection and infertility. A case-control study was conducted to retrospectively analyze the specimens submitted by patients from the Third Xiangya Hospital of Central South University from January 2021 to December 2022. The results showed that a total of 32 184 specimens were collected, and the positive rates of CT were 4.41% (1 419/32 184), and positive rats of NG were 1.42% (457/32 184). In the infertility group ( n=3 366), 2 987 were females and 379 were males. In the control group ( n=3 366), 2 509 were females and 857 were males. The CT positive rate of the infertility group was 13.61% (458/3 366), which was significantly higher than that of the control group 3.30% (111/3 366), and the difference was statistically significant ( χ2=4.245, P<0.05), and the NG positive rate of the infertility group was 6.36% (214/3 366), which was significantly higher than that of the control group 0.89% (30/3 366), and the difference was statistically significant ( χ2=4.011, P<0.05). A total of 23 992 female genital tract swab specimens were collected, including 2 987 in the infertility group and 2 509 in the control group, and the positive rate of CT in the female infertility subgroup was 10.41% (311/2 987), which was significantly higher than that in the control group 3.75% (94/2 509), the difference was statistically significant ( χ2=4.132, P<0.05), and the NG positive rate of 8.73% (261/2 987) in the female infertility subgroup was significantly higher than that in the control group 0.40% (10/2 509), and the difference was statistically significant ( χ2=4.242, P<0.05). A total of 8 192 male urine samples were collected, including 379 in the infertility group and 857 in the control group, and the CT positive rate of the male infertility subgroup was 13.72% (52/379), which was significantly higher than that of the control group 3.38% (29/857), and the difference was statistically significant ( χ2=5.267, P<0.05), and the positive rate of NG in the male infertility subgroup was 12.66% (48/379), which was significantly higher than that of the control group 0.93% (8/857), and the difference was statistically significant ( χ2=4.166, P<0.05). Among the 2 987 female specimens in the infertility group, 1 034 were in the primary infertility subgroup and 1 953 were in the secondary infertility subgroup, and the positive rates of CT were 7.93% (82/1 034) and 15.72% (307/1 953), respectively, and the positive rates of NG were 3.87% (40/1 034) and 8.65% (169/1 953) respectively, and the difference was not statistically significant ( χ2=0.185, P>0.05) and ( χ2=0.002, P>0.05). In conclusion, the infection rate of genital tract CT and NG is high in the infertility population, CT and NG are recommended as routine examination indicators for eugenics and infertility screening.

To analyze the infection of chlamydia (CT) and gonorrhea (NG) in female infertility and male infertility population, and to explore the correlation between CT and NG infection and infertility. A case-control study was conducted to retrospectively analyze the specimens submitted by patients from the Third Xiangya Hospital of Central South University from January 2021 to December 2022. The results showed that a total of 32 184 specimens were collected, and the positive rates of CT were 4.41% (1 419/32 184), and positive rats of NG were 1.42% (457/32 184). In the infertility group ( n=3 366), 2 987 were females and 379 were males. In the control group ( n=3 366), 2 509 were females and 857 were males. The CT positive rate of the infertility group was 13.61% (458/3 366), which was significantly higher than that of the control group 3.30% (111/3 366), and the difference was statistically significant ( χ2=4.245, P<0.05), and the NG positive rate of the infertility group was 6.36% (214/3 366), which was significantly higher than that of the control group 0.89% (30/3 366), and the difference was statistically significant ( χ2=4.011, P<0.05). A total of 23 992 female genital tract swab specimens were collected, including 2 987 in the infertility group and 2 509 in the control group, and the positive rate of CT in the female infertility subgroup was 10.41% (311/2 987), which was significantly higher than that in the control group 3.75% (94/2 509), the difference was statistically significant ( χ2=4.132, P<0.05), and the NG positive rate of 8.73% (261/2 987) in the female infertility subgroup was significantly higher than that in the control group 0.40% (10/2 509), and the difference was statistically significant ( χ2=4.242, P<0.05). A total of 8 192 male urine samples were collected, including 379 in the infertility group and 857 in the control group, and the CT positive rate of the male infertility subgroup was 13.72% (52/379), which was significantly higher than that of the control group 3.38% (29/857), and the difference was statistically significant ( χ2=5.267, P<0.05), and the positive rate of NG in the male infertility subgroup was 12.66% (48/379), which was significantly higher than that of the control group 0.93% (8/857), and the difference was statistically significant ( χ2=4.166, P<0.05). Among the 2 987 female specimens in the infertility group, 1 034 were in the primary infertility subgroup and 1 953 were in the secondary infertility subgroup, and the positive rates of CT were 7.93% (82/1 034) and 15.72% (307/1 953), respectively, and the positive rates of NG were 3.87% (40/1 034) and 8.65% (169/1 953) respectively, and the difference was not statistically significant ( χ2=0.185, P>0.05) and ( χ2=0.002, P>0.05). In conclusion, the infection rate of genital tract CT and NG is high in the infertility population, CT and NG are recommended as routine examination indicators for eugenics and infertility screening.

OBJECTIVE To systematically review the pharmacoeconomic evaluation literature about proprotein convertase subtilisin/kexin type 9 （PCSK9） inhibitors for the prevention of cardiovascular disease in patients with hypercholesterolemia， and to provide a reference for clinical treatment， health decision-making and future follow-up research. METHODS Retrieved from English and Chinese databases such as PubMed and CNKI， the pharmacoeconomic literature about PCSK9 inhibitors （evolocumab and alirocumab） for the prevention of cardiovascular disease in patients with hypercholesterolemia was collected from the establishment of the database to October 8， 2023. The quality of the included literature was assessed with Consolidated Health Economic Evaluation Reporting Standards 2022 （CHEERS 2022） scale. The descriptive analysis was performed for basic information， model structure， related parameters， sensitivity analysis and the results of included studies. RESULTS & CONCLUSIONS A total of 29 literature were included， with overall good quality. The evaluation mainly adopted the Markov model from the healthcare system， payer and societal perspective. The effectiveness and utility data mainly came from the previous studies； the direct cost was mainly considered with a discount rate of 1.5%-5.0% per year， while the willingness-to-pay threshold was often set at 1-3 times the gross domestic product per capita. Most health output indicators were measured in life years and quality-adjusted life years. The sensitivity analysis of most studies demonstrated the robustness and the main influential factor was the drug cost. Most Chinese studies showed that PCSK9 inhibitor was not cost-effective for the prevention of cardiovascular disease in patients with acute coronary syndrome， myocardial infarction and atherosclerotic cardiovascular disease. It was cost-effective to use PCSK9 inhibitors for the prevention of cardiovascular disease only in specific groups， such as patients with triple vessel disease， patients with newly diagnosed acute coronary syndrome and low-density lipoprotein cholesterol≥100 mg/dL. Future research should refer to the CHEERS 2022 scale to improve the design， and strive to select large-scale， high-quality data to enhance the quality of reports and the transparency of health decisions， so as to more accurately assess the cost-effectiveness of PCSK9 inhibitors.

Objective To construct a stable synovial cell line MH7A from rheumatoid arthritis(RA)patients using lentiviral vectors that interfere with the expression of tumor necrosis factor receptor associated factor 2(TRAF2),and to study the role of TNF-α-TRAF2 signaling in MH7A abnormal proliferation.Methods Based on the design principles of human TRAF2 gene sequence and shRNA sequence,three pairs of TRAF2 shRNA interference se-quences were designed and synthesized.The primers were annealed by PCR,and a linear vector was obtained by double enzyme digestion PLKO.1-puro.The linearized vector was connected to the annealed primers through Solu-tion I,and the connected products were introduced into receptive cells.The plates were coated,and positive colo-nies were selected for sequencing.Three different recombinant plasmids of PLKO.1-TRAF2-shRNA lentivirus were constructed,and lentivirus packaging plasmids was used to package logarithmic growth phase HEK 293T cells.Vi-rus solution was collected to infect MH7A cells.At the same time,puromycin was used to screen MH7A stable transgenic strains with low TRAF2 expression.CCK-8 method,Western blot,and qPCR were used to detect the proliferation function of MH7A induced by TNF-α and low expression of TRAF2,as well as downstream signal TRAF2,P65 protein expression and mRNA levels.Results PLKO.1-TRAF2-shRNA(1),PLKO.1-TRAF2-shR-NA(2),and PLKO.1-TRAF2-shRNA(3)lentivirus vector plasmids and control group lentivirus vector plasmids PLKO.1-puro were successfully constructed.The three TRAF2-shRNA lentivirus vector plasmids and control group lentivirus vector plasmids PLKO.1-puro were respectively introduced into the lentivirus packaging plasmid of HEK 293T to obtain virus solution.After infecting MH7A cells with the virus solution,they were treated with puromycin(2.00 μ G/mL)screening and obtaining MH7A stable transgenic plants after 2 days.Through qPCR and Western blot results,it was found that the expression of TRAF2 mRNA and protein in PLKO.1-TRAF2-shRNA(1)MH7A stably transfected cells was significantly reduced compared to the negative control group.The results of CCK-8 and Western blot showed that after knocking down TRAF2 in MH7A,the proliferation of MH7A cells with low TRAF2 expression induced by TNF-α and the phosphorylation level of P65 were significantly reduced.Conclusion A sta-ble transgenic strain of PLKO.1-TRAF2-shRNA(1)MH7A cells was successfully constructed to investigate the role of TNF-α-TRAF2 signal activation in mediating abnormal proliferation of RA synovial cells.

In 2023，World Health Organization（WHO）announced that the pandemic of COVID-19 is over；while there was an epidemic of multi-pathogenic infections in the respiratory tract by the end of the year in China，indicating that high attention still need to be paid to infectious diseases. In the past one year，some breakthroughs for infectious diseases have been achieved.This article reviews the major events and research progress in the diagnosis，prevention and treatment of infectious diseases in 2023.

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.

There is currently a huge worldwide demand for donor kidneys for organ transplantation. Consequently, numerous marginal donor kidneys, such as kidneys with microthrombi, are used to save patients' lives. While some studies have shown an association between the presence of microthrombi in donor kidneys and an increased risk for delayed graft function (DGF) (McCall et al., 2003; Gao et al., 2019), other studies have demonstrated that microthrombi negatively impact the rate of DGF (Batra et al., 2016; Hansen et al., 2018), but not graft survival rate (McCall et al., 2003; Batra et al., 2016; Gao et al., 2019). In contrast, Hansen et al. (2018) concluded that fibrin thrombi were not only associated with reduced graft function six months post-transplantation but also with increased graft loss within the first year of transplantation. On the other hand, Batra et al. (2016) found no significant differences in the DGF rate or one-year graft function between recipients in diffuse and focal microthrombi groups. To date, however, the overall influence of donor kidney microthrombi and the degree of influence on prognosis remain controversial, necessitating further research.
Humans ; Thrombotic Microangiopathies ; Transplantation, Homologous ; Tissue Donors ; Kidney ; Allografts