Objective To prepare rabbit antibody against mouse AD-004 by AD-004 expressed in the prokaryotic expression system and to identify its distribution in the testis and adrenal. Methods The full-length cDNA of mouse AD-004 was cloned into PET28 plasmid, and the protein was induced in E. coli BL21 bacteria by adding IPTC and then purified by Ni2+ -NTA column. The purified protein was used as an immunogen to prepare polyclonal antibody ( pAb) of AD-004. The specificity of the antibody was detected by Western blotting. Immunohistochemical staining was performed in the mouse adrenal and testis via pAb of AD-004. Results Hisfused AD-004 was expressed efficiently in the prokaryotic system. Western blot analysis showed that the polyclonal antibody was duly bound to purified AD-004 with high specificity and sensitivity. AD-004 could be abundantly identified in the adrenal medulla and mainly expressed in the Leydig cells of testis. Conclusion The mouse protein of AD-004 is obtained from the prokaryotic expression system. The rabbit anti-AD-004 antibody has been prepared successfully. AD-004 protein is mainly localized in the interstitium of testis, suggesting that AD-004 may play a role in the synthesis of sex-steroid hormone.

Objective To investigate the situation of sexual partners and related factors among men who have sex with men (MSM) in college students.Methods Snowball sampling and Convenience sampling were both used to recruit MSM from colleges in Tsingtao in 2016.Questionnaire-based interviews were conducted to collect data of socio-demographic and situation of sexual partners.Sample Size was estimated based on cross-sectional study,and theoretical 267.SPSS 17.0 software was used for statistical analysis.Results A total of 300 MSM,average aged 20.7,were analyzed.Both first sex partner and the last same-sex sexual partner were mct instantly,with proportions as 58.7％ (176/300) and 62.3％ (187/300) respectively.Among all the MSM,88.3％ (265/300) preferred selecting men as sex partners and 42.7％ (128/300) enjoyed finding sex partners in college,while 86.0％ (258/300) preferred finding their sex partners through intemet.Conclusions Intemet had been the major way of looking for sex partner among MSM in college students,the male sexual partner were met instantly.We should focus on men who have sex with men and their sexual health among college students to prevent and control HIV/AIDS.

Objective To study the differences in the blood cell analysis of male officers and soldiers be-tween in the high altitude area station and low altitude area station in summer.Methods A total of 239 male officers and soldiers in the high altitude area(Amdo Xizang,average altitude 4 800 m)and 336 male officers and soldiers in the low altitude area(Nanjing,Jiangsu,average altitude 30 m)from July 18 to 24,2022 were selected as the study subjects and the differences in blood cell analysis parameters of male officers and soldiers stationed between at high altitude and low altitude areas were retrospectively analyzed.Results The eosino-phils percentage(EDS％),eosinophils count(EOS)in the high altitude group were significantly lower than thosein the low altitude group(P＜0.05),and the basophillic granulocyte percentage(BASO％),basophillic granulo-cyte count(BASO)and monocyte percentage(MONO％)were significantly higher than those in the low alti-tude group,and the differences were statistically significant(P＜0.05),but which in the both groups were in the normal reference ranges.The red blood cell count(RBC)hemoglobin(Hb)and hematocrit(HCT)in the high altitude group were significantly higher than those in the low altitude group(P＜0.05),moreover Hb and HCT in the high altitude group were in the upper limit of the medical reference range.The mean corpus-cular volume(MCV),mean corpuscular hemoglobin(MCH),mean corpuscular hemoglobin concentration(MCHC)and red blood cell distribution width-standard diviation(RDW-SD)in the high altitude group were lower than those in the low-altitude group(P＜0.05),but the both groups were in the normal reference ran-ges;there was no statistically significant difference in the erythrocyte distribution width coefficient of variation(RDW-CV)between the two groups(P＞0.05).The platelet(PLT)and thrombocytocrit(PCT)in the high altitude group were higher than those in the low altitude group,the platelet distribution width(PDW),mean platelet volume(MPV)and platelet large cell ratio(P-LCR)were lower than those in the low altitude group,and the differences were statistically significant(P＜0.05);PDW in the low-altitude group was at the upper limit of the medical reference range,and the other platelet-related indexes were in the normal range.Conclusion There are obvious differences in the blood cell analysis indicators of male officers and soldiers be-tween the high altitude area and low altitude area.

BACKGROUND:Tyrosine kinase inhibitors,as well as other types of small-molecule cancer drugs,can cause severe cardiotoxicity. OBJECTIVE:To perform a heart safety re-evaluation by observing the effects of antitumor drugs on isolated heart electrocardiograph,cardiac action potential and associated ion channels and cytotoxicity. METHODS:Extracorporeal cardiac perfusion was given to the isolated rabbit heart using Langendorff perfusion:Sunitinib(0.3,3,10 μmol/L),Crizotinib(0.3,1,3 μmol/L),and Doxorubicin(1,30 μmol/L)were perfused sequentially for 120 minutes to record electrocardiograph and left ventricular pressure.A blank control group was set for comparison.Manual patch clamp was used to record the effects of Crizotinib,Sunitinib,Doxorubicin on hERG,Cav1.2,Nav1.5 channel currents and action potential in human induced pluripotent stem cell derived cardiomyocytes.Adenosine triphosphate level in human induced pluripotent stem cell derived cardiomyocytes was detected by CellTiter-Glo luminescent cell viability assay. RESULTS AND CONCLUSION:Isolated rabbit heart using Langendorff perfusion:Compared with the blank ontrol group,Sunitinib and Crizotinib at≥3 μmol/L decreased heart rate(P<0.01)and prolonged QT/QTc interval(P<0.01),and reduced left ventricular pressure to different extents.Manual patch clamp recording:Compared with the blank control group,Sunitinib and Crizotinib at 3 μmol/L inhibited the activities of hERG,Nav1.5 and Cav1.2 channels and significantly prolonged the duration of action potential(P<0.01).According to the analysis of the test article,the difference between the labeled concentration and the measured concentration of the recovered solution was not significant.Cell viability assays:Compared with the blank control group,adenosine triphosphate content in human induced pluripotent stem cell derived cardiomyocytes significantly decreased after treatment with Sunitinib(IC50=4.64 μmol/L),Doxorubicin(IC50=4.21 μmol/L)and Crizotinib(IC50=2.87 μmol/L),indicating that cell viability significantly decreased(P<0.01).To conclude,this study successfully established an early cardiac safety evaluation method for antitumor drugs,which provides good support and help for the subsequent development of antitumor drugs.

The continuing challenges that limit effectiveness of tumor therapeutic vaccines were high heterogeneity of tumor immunogenicity, low bioactivity of antigens, as well as insufficient lymph nodes (LNs) drainage of antigens and adjuvants. Transportation of in situ neoantigens and adjuvants to LNs may be an effective approach to solve the abovementioned problems. Therefore, an FA-TSL/AuNCs/SV nanoplatform was constructed by integrating simvastatin (SV) adjuvant loaded Au nanocages (AuNCs) as cores (AuNCs/SV) and folic acid modified thermal-sensitive liposomes (FA-TSL) as shells to enhance de novo antitumor immunity. After accumulation in tumor guided by FA, AuNCs mediated photothermal therapy (PTT) induced the release of tumor-derived protein antigens (TDPAs) and the shedding of FA-TSL. Exposed AuNCs/SV soon captured TDPAs to form in situ recombinant vaccine (AuNCs/SV/TDPAs). Subsequently, AuNCs/SV/TDPAs could efficiently transport to draining LNs owing to the hyperthermia induced vasodilation effect and small particle size, achieving co-delivery of antigens and adjuvant for initiation of specific T cell response. In melanoma bearing mice, FA-TSL/AuNCs/SV and laser irradiation effectively ablated primary tumor, against metastatic tumors and induced immunological memory. This approach served a hyperthermia enhanced platform drainage to enable robust personalized cancer vaccination.

Activating humoral and cellular immunity in lymph nodes (LNs) of nanoparticle-based vaccines is critical to controlling tumors. However, how the physical properties of nanovaccine carriers orchestrate antigen capture, lymphatic delivery, antigen presentation and immune response in LNs is largely unclear. Here, we manufactured gold nanoparticles (AuNPs) with the same size but different shapes (cages, rods, and stars), and loaded tumor antigen as nanovaccines to explore their disparate characters on above four areas. Results revealed that star-shaped AuNPs captured and retained more repetitive antigen epitopes. On lymphatic delivery, both rods and star-shaped nanovaccines mainly drain into the LN follicles region while cage-shaped showed stronger paracortex retention. A surprising finding is that the star-shaped nanovaccines elicited potent humoral immunity, which is mediated by CD4⁺ T helper cell and follicle B cell cooperation significantly preventing tumor growth in the prophylactic study. Interestingly, cage-shaped nanovaccines preferentially presented peptide-MHC I complexes to evoke robust CD8⁺ T cell immunity and showed the strongest therapeutic efficacy when combined with the PD-1 checkpoint inhibitor in established tumor study. These results highlight the importance of nanoparticle shape on antigen delivery and presentation for immune response in LNs, and our findings support the notion that different design strategies are required for prophylactic and therapeutic vaccines.