GPR126, also known as ADGRG6, is one of the most deeply studied aGPCRs. Initially, GPR126 was thought to be a receptor associated with muscle development and was primarily expressed in the muscular and skeletal systems. With the deepening of research, it was found that GPR126 is expressed in multiple mammalian tissues and organs, and is involved in many biological processes such as embryonic development, nervous system development, and extracellular matrix interactions. Compared with other aGPCRs proteins, GPR126 has a longer N-terminal domain, which can bind to ligands one-to-one and one-to-many. Its N-terminus contains five domains, a CUB (complement C1r/C1s, Uegf, Bmp1) domain, a PTX (Pentraxin) domain, a SEA (Sperm protein, Enterokinase, and Agrin) domain, a hormone binding (HormR) domain, and a conserved GAIN domain. The GAIN domain has a self-shearing function, which is essential for the maturation, stability, transport and function of aGPCRs. Different SEA domains constitute different GPR126 isomers, which can regulate the activation and closure of downstream signaling pathways through conformational changes. GPR126 has a typical aGPCRs seven-transmembrane helical structure, which can be coupled to Gs and Gi, causing cAMP to up- or down-regulation, mediating transmembrane signaling and participating in the regulation of cell proliferation, differentiation and migration. GPR126 is activated in a tethered-stalk peptide agonism or orthosteric agonism, which is mainly manifested by self-proteolysis or conformational changes in the GAIN domain, which mediates the rapid activation or closure of downstream pathways by tethered agonists. In addition to the tethered short stem peptide activation mode, GPR126 also has another allosteric agonism or tunable agonism mode, which is specifically expressed as the GAIN domain does not have self-shearing function in the physiological state, NTF and CTF always maintain the binding state, and the NTF binds to the ligand to cause conformational changes of the receptor, which somehow transmits signals to the GAIN domain in a spatial structure. The GAIN domain can cause the 7TM domain to produce an activated or inhibited signal for signal transduction, For example, type IV collagen interacts with the CUB and PTX domains of GPR126 to activate GPR126 downstream signal transduction. GPR126 has homology of 51.6%-86.9% among different species, with 10 conserved regions between different species, which can be traced back to the oldest metazoans as well as unicellular animals.In terms of diseases, GPR126 dysfunction involves the pathological process of bone, myelin, embryo and other related diseases, and is also closely related to the occurrence and development of malignant tumors such as breast cancer and colon cancer. However, the biological function of GPR126 in various diseases and its potential as a therapeutic target still needs further research. This paper focuses on the structure, interspecies differences and conservatism, signal transduction and biological functions of GPR126, which provides ideas and references for future research on GPR126.

Objective To explore feasibility of 3D metal printing technology combined with virtual design proximal clavicle anatomical plate.Methods A 52-year-old male healthy volunteer was retrospectively selected to design proximal clavicle anatomical plate system by using Mimics15.01,NX12.0 and other software.STL data were input into 3D printer to print 1∶1 clavicle model and proximal clavicle anatomical plate.The fit of the plate was tested in vitro and the accuracy of screw position was evaluated by imaging.Printing time of model,nail path design and fabrication time of the anatomical plate at proximal clavicle were recorded.Results The 3D metal printing proximal clavicle anatomical plate fitted well to clavicle model,orienta-tion of proximal clavicle locking screw was accurate,and X-ray and CT scan showed the screw position was good.Printing time of model,the time of nail path design,and the time of making anatomical plate of proximal clavicle were 120,15 and 300 min respectively.Conclusion The proximal clavicular anatomical plate system based on 3D metal printing technology could achieve good lamination of proximal clavicular fracture plate and precise screw placement,providing a new and accurate surgical method for the treatment of the proximal clavicular fracture.

This study was aimed at analyzing the molecular epidemiological characteristics of all 14 cases of human infection with avian influenza A(H7N9)virus in Xinjiang from 2014 to 2018,to provide a scientific basis for prevention,control,and treatment.The genomic sequence was obtained through high-throughput gene sequencing after nucleic acid extraction.Homolo-gy analysis,evolution analysis,mutation locus analysis,and homology modeling were performed in bioinformatics analysis software.The nucleotide homology and amino acid homology of the HA gene in 14 human infected H7N9 viruses were(97.39％-100％)and(98.38％-100％),respectively.The nucleotide homology of the NA gene and the amino acid homology ranged from 97.73％to 100％.All viruses were low pathogenic avian influenza viruses belonging to the Yangtze River Delta lin-eage and were divided into two subclades,which were most similar to the A/Hunan/02650/2016 vaccine strain.All HA pro-teins G186V and T160A were mutated;13 strains of Q226L were mutated;and none of the four key neuraminidase inhibitor resistance sites of NA protein were mutated.All sites of M2 protein S31N and V27A were mutated,all sites of PB1 protein T368V were mutated,and all sites of PA protein K356R were mutated.Xinjiang H7N9 virus exhibited double receptor bind-ing,and was resistant to amantadine drugs and sensitive to neuraminidase inhibitors,which may be used in early disease sta-ges.Strengthened monitoring and timely detection of avian in-fluenza virus genome changes will be critical for prevention and control,and formulation of countermeasures.

The spleen is the largest lymphoid organ of the body,which participates in the regulation of metabolic balance.High altitude hypoxia environment can affect lipid metabolism in spleen tissue,but the key mechanism of lipid metabolism is still unclear.We aimed to use lipidomic analysis to study the effect of high altitude hypoxia on lipid metabolism in mouse spleen tissue.C57BL/6 mice were placed at an altitude of 4 200 m and 400 m,respectively,and after 30 days the spleen tissues were harvested and lipidomic analysis was performed using an ultra-high performance liquid chromatography-Orbitrap mass spectrometry system.Under the high altitude hypoxia environment,the spleen index of mice and the white pulp decreased,and the germinal center expanded with other pathological changes.The results of lipidomic analysis showed that a total of 41 lipid subclasses and 2 473 lipid molecules were identified,and triacylglycerides(TGs)and phosphatidylcholines(PCs)were the two most identified lipid mole-cules.Using univariate and multivariate analysis,44 differentially expressed lipid molecules were identi-fied,which were mainly concentrated in phospholipid metabolism.Subsequently,RT-qPCR was per-formed on the key enzymes in the phospholipid metabolic pathway,and it was found that the mRNA ex-pression levels were different(P＜0.05).It suggested that high altitude hypoxia environment mainly af-fects the phospholipid metabolism of mouse spleen tissue via reducing the contents of PC and phosphatidic acid(PA),promoting their conversion to phosphatidylethanolamine(PE)and cardiolipin(CL)and fa-cilitating the PE production via the CDP-Etn pathway.This study provides a new experimental basis for the abnormal metabolism of phospholipids in spleen tissue under high altitude hypoxia environment.

Objective:To investigate the toxic effect of chlorambucil combined with ibrutinib on mantle cell lymphoma(MCL)cell line Jeko-1 and its related mechanism.Methods:The MCL cell line Jeko-1 was incubated with different concentrations of chlorambucil or ibrutinib or the combination of the two drugs,respectively.CCK-8 assay was used to detect the proliferation of the cells,and Western blot was used to measure the protein expression levels of BCL-2,caspase-3,PI3K,AKT and P-AKT.Results:After Jeko-1 cells were treated with chlorambucil(3.125,6.25,12.5,25,50 μmol/L)and ibrutinib(3.125,6.25,12.5,25,50 μmol/L)alone for 24,48,72h respectively,the cell proliferation was inhibited in a time-and dose-dependent manner.Moreover,the two drugs were applied in combination at low doses(single drug inhibition rate＜50％),and the results showed that the combination of two drugs had a more significant inhibitory effect(all P＜0.05).Compared with the control group,the apoptosis rate of the single drug group of chlorambucil(3.125,6.25,12.5,25,50 μmol/L)and ibutinib(3.125,6.25,12.5,25,50 μmol/L)was increased in a dose-dependent manner.The combination of the two drugs at low concentrations(3.125,6.25,12.5 μmol/L)could significantly increase the apoptosis rate compared with the corresponding concentration of single drug groups(all P＜0.05).Compared with control group,the protein expression levels of caspase-3 in Jeko-l cells were upregulated,while the protein expression levels of BCL-2,PI3K,and p-AKT/AKT were downregulated after treatment with chlorambucil or ibrutinib alone.The combination of the two drugs could produce a synergistic effect on the expressions of the above-mentioned proteins,and the differences between the combination group and the single drug groups were statistically significant(all P＜0.05).Conclusion:Chlorambucil and ibrutinib can promote the apoptosis of MCL cell line Jeko-1,and combined application of the two drugs shows a synergistic effect,the mechanism may be associated with the AKT-related signaling pathways.

Objective:To investigate the clinical efficacy and safety of ixazomib-containing regimens in the treatment of patients with multiple myeloma(MM).Methods:A retrospective analysis was performed on the clinical efficacy and adverse reactions of 32 MM patients treated with a combined regimen containing ixazomib in the Hematology Department of the First People's Hospital of Lianyungang from January 2020 to February 2022.Among the 32 patients,15 patients were relapsed and refractory multiple myeloma(R/RMM)(R/RMM group),17 patients who responded to bortezomib induction therapy but converted to ixazomib-containing regimen due to adverse events(AE)or other reasons(conversion treatment group).The treatment included IPD regimen(ixazomib+pomalidomide+dexamethasone),IRD regimen(ixazomib+lenalidomide+dexamethasone),ICD regimen(ixazomib+cyclophosphamide+dexamethasone),ID regimen(ixazomib+dexamethasone).Results:Of 15 R/RMM patients,overall response rate(ORR)was 53.3％(8/15),among them,1 achieved complete response(CR),2 achieved very good partial response(VGPR)and 5 achieved partial response(PR).The ORR of the IPD,IRD,ICD and ID regimen group were 100％(3/3),42.9％(3/7),33.3％(1/3),50％(1/2),respectively,there was no statistically significant difference in ORR between four groups(x2=3.375,P=0.452).The ORR of patients was 50％after first-line therapy,42.9％after second line therapy,60％after third line therapy or more,with no statistically significant difference among them(x2=2.164,P=0.730).In conversion treatment group,ORR was 88.2％(15/17),among them,6 patients achieved CR,5 patients achieved VGPR and 4 patients achieved PR.There was no statistically significant difference in ORR between the IPD(100％,3/3),IRD(100％,6/6),ICD(100％,3/3)and ID(60％,3/5)regimen groups(x2=3.737,P=0.184).The median progression-free survival(PFS)time of R/RMM patients was 9 months(95％CI:6.6-11.4 months),the median overall survival(OS)time was 18 months(95％CI:11.8-24.4 months).The median PFS time of conversion treatment group was 15 months(95％CI:7.3-22.7 months),the median OS time not reached.A total of 10 patients suffered grade 3-4 adverse event(AE).The common hematological toxicities were leukocytopenia,anemia,thrombocytopenia.The common non-hematological toxicities were gastrointestinal symptoms(diarrhea,nausea and vomit),peripheral neuropathy,fatigue and infections.Grade 1-2 peripheral neurotoxicity occurred in 7 patients.Conclusion:The ixazomib-based chemotherapy regimens are safe and effective in R/RMM therapy,particularly for conversion patients who are effective for bortezomib therapy.The AE was manageable and safe.

Objective:To screen interleukin(IL)-1β secretion-related membrane transporters by macrophage experiment in vitro and conventional knockout mice.Methods:THP-1 cell line was differentiated to obtain human THP-1-derived macrophages,and the primary macrophages were obtained from human peripheral blood.FVB wild-type mice with the same sex and age were used as the controls of MRP1 knockout mice.The macrophages in abdominal cavity and bone marrow of mice were cultivated.The cells were treated with ABCC1/MRP1,ABCG2/BCRP,ABCB1/P-gp,OATP1B1,and MATE transporter inhibitors,then stimulated by lipopolysaccharide and adenosine triphosphate.The secretion level of IL-iβ was detected by ELISA,Western blot,and immunofluorescence.Results:After inhibiting ABCC1/MRP1 transporter,the secretion of IL-1β decreased significantly,while inhibition of the other 4 transporters had no effect.In animal experiment,the level of IL-1 β secreted by macrophages in bone marrow of MRP1 knockout mice was significantly lower than control group(P＜0.05).Conclusion:ABCC1/MRP1 transporter is a newly discovered IL-1β secretion pathway,which is expected to become a new target for solving clinical problems such as cytokine release syndrome.

Objective:To detect the pharmacokinetic(PK)parameters of coagulation factor Ⅷ(FⅧ)in adult patients with severe hemophilia A,identify the potential factors influencing FⅧ PK,and optimize the use of FⅧ in individual prophylaxis regimens.Methods:PK characteristics of FⅧ were studied in a total of 23 severe hemophilia A adults.The correlation of patients'characteristics including age,von Willebrand factor antigen(vWF:Ag),blood group,weight,body mass index(BMI)and FⅧ genotype,with FⅧ PK were evaluated.Individual prophylaxis regimens were given based on FⅧ PK parameters.Results:The mean terminal half-life(t1/2)of FⅧ was 20.6±9.3 h,ranged from 11.47 h to 30.12 h.The age(r=0.580)and vWF:Ag(r=0.814)were significantly positively correlated with t1/2 of FⅧ.The mean area under the plasma concentration curve(AUC)of FⅧ was 913±399(328-1 878)IU h/dl,and the AUC of FⅧ was positively correlated with age(r=0.557)and vWF:Ag(r=0.784).The mean residence time(MRT)of FⅧ was 24.7±12.4(13.2-62.2)h,and the MRT of FⅧ was positively correlated with age(r=0.664)and vWF:Ag(r=0.868).The mean in vivo recovery(IVR)of FⅧ was 2.59±0.888(1.5-4.29)IU/dl per IU/kg,the mean clearance(CL)of FⅧ was 3±1.58(0.97-7.18)ml/(kg·h),and there was no significant correlation of IVR and CL with age and vWF:Ag.According to the individual PK parameters,ultra low-dose,low-dose and moderate-dose FⅧ were applied to 15,6,2 adults patients with severe hemophilia A for prophylaxis,respectively.Conclusion:There are significant individual differences in the FⅧ half-life of adult patients with severe hemophilia A.The older the patient,the higher the vWF:Ag level,and the longer the FⅧ half-life.Individual administration is required based on the FⅧ PK parameters to optimize prophylaxis treatment.

Objective:To investigate the effect of feeder layer cells expressing interleukin(IL)-21 on the amplification of NK cells in vitro.Methods:The K562 cell line with IL-21 expression on its membrane was constructed by electroporation,and co-cultured with NK cells after inactivation.The proliferation of NK cells was observed.The killing function of the amplified NK cells in vitro was evaluated by the lactate dehydrogenase(LDH)and interferon-γ(IFN-y)release assay.A colorectal cancer xenograft model in NOD/SCID mice was established,and a blank control group,a NK cell group and an amplified NK cell group were set up to detect the tumor killing effect of amplified NK cells in vivo.Results:K562 cells expressing IL-21 on the membrane were successfully constructed by electroporation.After co-culturing with K562 cells expressing IL-21 on the membrane for 17 days,the NK cells increased to 700 times,which showed an enhanced amplification ability compared with control group(P＜0.001).In the tumor cell killing experiment in vitro,there was no significant difference in the killing activity on tumor cells between NK cells and amplified NK cells,and there was also no significant difference in mice in vivo.Conclusion:K562 cells expressing IL-21 on the membrane can significantly increase the amplification ability of NK cells in vitro,but do not affect the killing function of NK cells in vitro and in vivo.It can be used for the subsequent large-scale production of NK cells in vitro.

Objective To analyze the status of occupational hazards in key positions of the small and micro book and newspaper printing enterprises in Beijing City. Methods A total of 16 small and micro printing enterprises in Beijing City were selected as the research subjects using the judgment sampling method. The status of occupational hazard was assessed using on-site occupational health investigations. The volatile chemical components in organic solvents such as inks and cleaning agents used in the workplace, the level of chemical hazardous agents in the air, and noise intensity were detected and analyzed. Results A total of 1 105 workers from 16 small and micro printing enterprises were included. The occupational hazard exposure rate was 22.2% (245/1 105), with chemical hazardous agents and noise exposure rates of 13.5% and 22.2%, respectively. The rate of compliance with occupational health training among the head of the enterprise and the occupational health management personnel, the regular detection rate of occupational hazards in last year, and the rate for the occupational health examination in last year were both 100.0%. The rate for annual update of occupational hazard project reporting was 93.8% (15/16). The usage rates of gas mask and noise-proof earplug were 68.8% (11/16) and 50.0% (8/16), respectively. The effectiveness rates of anti-toxic and noise-reduction facilities were 87.5% (14/16) and 62.5% (10/16). The sign setting rates of chemical hazardous agents and noise warning were 93.8% (15/16). Acetone and isopropanol were found in ink and cleaning agents in 15 printing enterprises, while toluene, xylene, and ethylbenzene were found in three, two, and one enterprise, respectively. Benzene, 1,2-dichloroethane, n-hexane, and trichloroethylene were not found in all 16 enterprises. Both the exposure concentration of short term and exposure concentration of time weighted average of the above chemical hazards were lower than the lower limit of quantitation of the detection method in the workplace and work site air among the 16 printing enterprises, and none of concentration was exceeded the national standard. A total of 240 noise work sites were detected, and the national noise-exceeding rate was 11.2%(27/240). Conclusion Noise hazards are prominent in the small and micro printing enterprises in Beijing City, and attention should be paid to the prevention and protection of noise-exposure. The usage rate of personal protective equipment is not high, and the training and on-site supervision management of protective equipment wearing in workers should be strengthened.