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.

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 combined orthodontic-orthognathic treatment, the maxillary palatine suture is closed in most patients with insufficient maxillary width, and bony expansion of the maxilla cannot be achieved by dental expansion or rapid palatal expansion (RPE) which causes buccal inclination of the maxillary posterior teeth leading to unstable results. Therefore, segmental LeFort Ⅰ osteotomy and surgically assisted RPE are often used in clinical practice. In recent years, with the application of implant anchorage technology, implant anchorage assisted RPE has been gradually applied in orthognathic treatment. This article reviewed the indications, contraindications, complications, efficacy and long-term stability in different treatment approaches including segmental LeFort Ⅰ osteotomy, surgically assisted RPE and implant-supported maxillary skeletal expansion.

The nomenclature of congenital auricular malformations is complex and confusing, which brings great inconvenience to clinical work and scientific research. This study comprehensively summarized and sorted out the existing Chinese and English names of congenital auricular malformations, providing reference basis for the scientific classification and clinical treatment of congenital auricular malformations. Sixty-seven papers on the topic of congenital auricle malformations were collected from databases such as CNKI, Wanfang Database, VIP Database, PubMed, Web of Science, and ScienceDirect by using the PRISMA method. The two core issues of morphological and anatomical naming and classification of congenital auricle malformations were sorted and summarized based on these articles. In this study, the existing Chinese and English names of congenital auricular malformations were summarized, and a preliminary classification was made at the morphological and anatomical levels, and the embryological origin of the external auricle and its substructures was elaborated. Based on the result of this study, a comprehensive analysis can be conducted on the malformations of the ear structures and their embryological origins in order to explore the etiology of congenital ear malformations. It is expected that scientific and standardized naming and classification method can be developed, and reasonable and effective standardized treatment can be formulated for congenital auricular malformations based on this study.

The nomenclature of congenital auricular malformations is complex and confusing, which brings great inconvenience to clinical work and scientific research. This study comprehensively summarized and sorted out the existing Chinese and English names of congenital auricular malformations, providing reference basis for the scientific classification and clinical treatment of congenital auricular malformations. Sixty-seven papers on the topic of congenital auricle malformations were collected from databases such as CNKI, Wanfang Database, VIP Database, PubMed, Web of Science, and ScienceDirect by using the PRISMA method. The two core issues of morphological and anatomical naming and classification of congenital auricle malformations were sorted and summarized based on these articles. In this study, the existing Chinese and English names of congenital auricular malformations were summarized, and a preliminary classification was made at the morphological and anatomical levels, and the embryological origin of the external auricle and its substructures was elaborated. Based on the result of this study, a comprehensive analysis can be conducted on the malformations of the ear structures and their embryological origins in order to explore the etiology of congenital ear malformations. It is expected that scientific and standardized naming and classification method can be developed, and reasonable and effective standardized treatment can be formulated for congenital auricular malformations based on this study.

Objective To investigate serum levels of soluble apoptosis-related factor ligand(sFasL)and stromal cell derived factor 1(SDF-1)in patients with aplastic anemia(AA)and their correlation with immunosuppressive treatment.Methods Forty-three AA patients who received immunosuppressive therapy for half a year were selected as the observation group,and another 43 healthy subjects at the same period were selected as the control group.Patients in the observation group received immunosuppressive therapy and hematopoietic propoietic therapy,and patients were divided into the ineffective group and the effective group according to the efficacy.Clinical data of patients were collected,and serum levels of sFasL,SDF-1,tumor necrosis factor-α(TNF-α)and interleukin-8(IL-8)were detected by enzyme-linked immunosorbent assay.The correlation between serum sFasL and SDF-1 levels was analyzed.Multivariate Logistic regression analysis was conducted to analyze influencing factors of immunosuppressive treatment in AA patients.The predictive values of serum sFasL and SDF-1 on immunosuppressive treatment were analyzed by receiver operating characteristic(ROC)curve.Results Serum levels of sFasL and SDF-1 were higher in the observation group than those in the control group(P＜0.05).Serum levels of TNF-α,IL-8,sFasL and SDF-1 were higher in the ineffective group than those in the effective group(P＜0.05).Serum sFasL level was positively correlated with SDF-1 level in AA patients(r=0.534,P＜0.001).Increased serum sFasL and SDF-1 levels were independent risk factors for ineffective immunosuppressive therapy in AA patients(P＜0.05).The area under the curve(AUC)of serum sFasL and SDF-1 combined to predict the immunosuppressive treatment effect of AA patients was 0.973(95%CI:0.872-0.999),which was better than that of single diagnosis(P＜0.05),and its sensitivity and specificity were 94.44%and 96.00%,respectively.Conclusion Serum levels of sFasL and SDF-1 are significantly increased in patients with AA.The combined detection of sFasL and SDF-1 has high predictive value for the immunosuppressive treatment effect of AA.

Objective:To investigate the clinical infection status and trends in drug resistance of a rare pathogen Kluyveromyces marxianus ( Candida kefyr), and to provide experience for clinical diagnosis and treatment. Methods:Morphological and molecular biological identification tests and in vitro microdilution drug susceptibility test were conducted on a Kluyveromyces marxianus strain recently isolated from the midstream urine sample of a patient with urinary calculus in the Fungal Laboratory, Changhai Hospital. The ultrastructural damage of the strain caused by different antifungal drugs was observed by scanning electron microscopy. A retrospective analysis was conducted on clinical cases of Kluyveromyces marxianus infection in Changhai Hospital from 2009 to 2021. Results:The isolated strain formed smooth, soft, cheese-like yeast colonies on the Sabouraud′s agar medium, and ovoid or slender spores were observed under the microscope. Morphological analysis, mass spectrometry and sequencing analysis identified the strain as Kluyveromyces marxianus. The drug susceptibility test showed that minimum inhibitory concentrations (MICs) of amphotericin B, fluconazole, itraconazole, voriconazole, posaconazole, flucytosine, caspofungin, and micafungin were 0.5, 0.5, 0.03, ≤ 0.03, 0.06, 0.5, ≤ 0.016, and 0.06 μg/ml, respectively. Under the scanning electron microscope, the strain was ovoid to slender before antifungal drug treatment, with a size of (3.0 - 6.5) μm × (5.5 - 11.0) μm; after 24-hour treatment with antifungal drugs at the dose of 1 μg/ml, cell membrane shrinkage was more obvious under the treatment with posaconazole, which exhibited a stronger destructive effect on the strain compared with amphotericin B and voriconazole. From 2009 to 2021, 8 cases of Kluyveromyces marxianus infection were collected, including 6 males and 2 females; the Kluyveromyces marxianus strains were isolated from ascites in 3 cases, bronchoalveolar lavage fluid in 1 case, sputum in 2 cases, and midstream urine samples in 2 cases. Conclusion:For suspected Kluyveromyces marxianus infection, it is crucial to determine the pathogenic species as early as possible using various identification methods, and to collect strain as well as evaluate drug susceptibility, which will be beneficial for targeted clinical treatment.

Objective:To understand the effect of vitamin B1 on lymphocyte function in simulated microgravity.Methods:Splenocytes of mice were isolated,and the rotatary cell culture system was used to simulate microgravity.Lymphocytes were stimulated with mitotic agents Concanavalin A,and cells were treated with different concentrations of vitamin B1,proliferation indexes of lympho-cytes and levels of cytokines in supernatant were detected.Results:Simulated microgravity could inhibit proliferation of splenic lym-phocytes,and decrease levels of cytokines,while vitamin B1 could promote lymphocyte proliferation and cytokines production in cells cultured in simulated microgravity in a dose dependent manner.Conclusion:Vitamin B1 may attenuate the inhibitory effect of simulated microgravity on lymphocytes by regulating cell proliferation and secretion of cytokines.

Objective This study aims to predict the coronavirus disease 2019(COVID-19)epidemic in Xi'an based on SEAIQR model and Dropout-LSTM model,and to provide a scientific basis for evaluating the effectiveness of the"dynamic zero-COVID policy".Methods Considering a large number of asymptomatic infections,the changing parameters,and control procedures,we developed a time-dependent susceptible-exposed-asymptomatic-infected-quarantined-removed(SEAIQR)model with stage-specific interventions.Considering the time-series characteristics of COVID-19 data and the nonlinear relationship between them,we constructed a deep learning Dropout-LSTM model.The data of newly confirmed cases in Xi'an from December 9th,2021 to January 31st,2022 were used to fit the model,and the data from February 1st,2022 to February 7th,2022 were used to evaluate the model performance of forecasting.We then calculated the effective reproduction number(Rt)and analyzed the sensitivity of the different measurement scenarios.Results The peak of newly confirmed cases predicted by the SEAIQR model would appear on December 26th,2021,with 176 cases,and the"dynamic zero-COVID policy"may be achieved in January 24th,2022,with R2=0.849.The Dropout-LSTM model can reflect the time-series and nonlinear characteristics of the data,and the predicted newly confirmed cases were highly consistent with the actual situation,with R2=0.937.The MAE and RMSE of the Dropout-LSTM model were lower than those of the SEAIQR model,indicating that the predicted results were more ideal.At the beginning of the outbreak,R0 was 5.63.Since the implementation of comprehensive control,Rt has shown a gradual downward trend,dropping to below 1.0 on December 27th,2021.With the reduction of effective contact rate,the early implementation of control measures and the improvement of immunity threshold,the peak of newly confirmed cases will continue to decrease.Conclusion The proposed Dropout-LSTM model forecasts the epidemic well,which can provide a reference for decision-making of the"dynamic zero-COVID policy."