Objective　To investigate the relationship between the degree of cervical lesions and the status of paired box-1 (PAX1) and tumor protein 63(TP63) gene promoter methylation in human papillomavirus (HPV)-positive patients with cervical lesions, as well as analyze their clinical significance. Methods Cervical tissue specimens were collected from 128 patients who were suspected of cervical lesions and HPV infection, and admitted to Qionghai People's Hospital between December 2021 and December 2022. According to pathological examination results, the patients were divided into the low-grade squamous intraepithelial lesion (LSIL) group (n=43), high-grade squamous intraepithelial lesion (HSIL) group (n=51) and cervical cancer group (n=34). The second-generation hybrid capture method was used for viral load. The degree of PAX1 and TP63 gene promoter methylation in each group was detected by bisulfite sequencing, and mRNA expression of PAX1 and TP63 was detected by real-time quantitative PCR. The diagnostic performance of the degree of PAX1 and TP63 methylation for cervical intraepithelial neoplasia (CIN2+) was evaluated. Results There were statistically significant differences in HR-HPV viral load between the groups (P>0.05). A total of 49 (38.28%) patients with PAX1 gene promoter methylation, and 55 (42.97%) patients with TP63 gene promoter methylation were detected among the 128 patients. The percentages of PAX1 and TP63 gene promoter methylation in the cervical cancer group, HSIL group and LSIL group were (67.65% and 73.53%), (43.14% and 49.02%) and (9.30% and 11.63%), with statistically significant differences between groups (P<0.05). The mRNA expression levels of PAX1 and TP63 in the cervical cancer group, HSIL group and LSIL group were [(0.34±0.08) and (0.45±0.13)], [(0.72±0.11) and (0.63±0.09)], [(1.04±0.09) and (0.87±0.11)], with statistically significant differences between groups (P<0.05). Receiver operating characteristic (ROC) curve analysis showed that the area under the curve (AUC) values of PAX1, TP63 gene promoter methylation, and their combination for diagnosing CIN2+ were 0.793, 0.842, and 0.857, respectively. The sensitivity values were 77.78%, 83.33%, and 77.78%. The specificity values were 80.85%, 85.11%, and 93.62%. The combined detection can improve the specificity of diagnosis of CIN2+ lesions. Conclusions The degree of PAX1 and TP63 gene promoter methylation is closely related to cervical lesions in patients with HPV infection, which indicates that it can be used as potential auxiliary indicators for the clinical diagnosis of CIN2+lesions.

Military pilot selection is an effective way to lower the training cost, ensure the safety of pilots and aircraft, and improve the military efficiency, which involves a comprehensive evaluation of medical fitness, psychology and flying aptitude of a pilot candidate.The concept and classification of comprehensive evaluation methods were introduced in this paper.Comprehensive evaluation methods used in domestic and overseas military pilot selection were reviewed.Finally, suggestions on the research and application of the military pilot selection system in China were raised.

BACKGROUND:Bone relies on the close connection between blood vessels and bone cells to maintain its integrity.Bones are in a physiologically hypoxic environment.Therefore,the study of angiogenesis and osteogenesis in hypoxic environment is closer to the microenvironment in vivo. OBJECTIVE:To explore the influence of Wenshen Tongluo Zhitong(WSTLZT)formula on H-type bone microvascular endothelial cell/bone marrow mesenchymal stem cell co-culture system in hypoxia environment and its related mechanism. METHODS:Enzyme digestion method and flow sorting technique were used to isolate and identify H-type bone microvascular endothelial cells.Mouse bone marrow mesenchymal stem cells were isolated and obtained by bone marrow adhesion method.H-type bone microvascular endothelial cell/bone marrow mesenchymal stem cell hypoxic co-culture system was established using Transwell chamber and anoxic culture workstation.WSTLZT formula powder was used to intervene in each group at a mass concentration of 50 and 100 μg/mL.The angiogenic function of H-type bone microvascular endothelial cells in the co-culture system was evaluated by scratch migration test and tube formation test.The osteogenic differentiation ability of bone marrow mesenchymal stem cells in the co-cultured system was evaluated by alkaline phosphatase staining and alizarin red staining.The protein and mRNA expression changes of PDGF/PI3K/AKT signal axis related molecules in H-type bone microvascular endothelial cells in the co-cultured system were detected by Western Blotting and q-PCR,respectively. RESULTS AND CONCLUSION:(1)Compared with the normal oxygen group,the scratch mobility and new blood vessel length of H-type bone microvascular endothelial cells were significantly higher(P<0.05);the osteogenic differentiation capacity of bone marrow mesenchymal stem cells was higher(P<0.05);the expression of PDGF/PI3K/AKT axis-related molecular protein and mRNA increased(P<0.05)in the hypoxia group.(2)Compared with the hypoxia group,scratch mobility and new blood vessel length were significantly increased in the H-type bone microvascular endothelial cells(P<0.05);bone marrow mesenchymal stem cells had stronger osteogenic function(P<0.05);the expression of PDGF/PI3K/AKT axis-related molecular proteins and mRNA further increased(P<0.05)after treatment with different dose concentrations of WSTLZT formula.These findings conclude that H-type angiogenesis and osteogenesis under hypoxia may be related to the PDGF/PI3K/AKT signaling axis,and WSTLZT formula may promote H-type vasculo-dependent bone formation by activating the PDGF/PI3K/AKT signaling axis,thereby preventing and treating osteoporosis.

OBJECTIVE To study the intervention effects of Wenshen Tongluo Zhitong Recipe(WTZR)on macrophage senes-cence and senile osteoporosis.METHODS The senescent macrophage model was established using hydrogen peroxide(H2 O2)and subsequently divided into four groups:control,model,low-dose drug-treated serum,and high-dose drug-treated serum.β-galactosidase staining,Western blot and qPCR were employed to evaluate the mRNA expression of senescence markers p21 and p53.ROS staining and JC-1 staining were applied to assess mitochondrial function in macrophages.The mRNA levels of Interleukin(IL)-6,IL-10,CD206,and inducible nitric oxide synthase(iNOS)were determined by qPCR analysis.Immunofluorescence was used to evaluate argi-nase(ARG1)and iNOS protein expressions for assessing the impact of drug-containing serum on macrophage polarization.qPCR analy-sis was conducted to measure osteocalcin(OCN),collagen type Ⅰ alpha 1(Col1a1),runt-related transcription factor 2(Runx2)mRNA levels as osteoblast-related markers;ALP staining along with alizarin red staining were performed to evaluate the effect of macrophage conditioned medium treated with drug-containing serum on bone marrow mesenchymal stromal cell(BMSC)osteogenic differentiation.C57BL/6J mice were randomly allocated into four groups:control group,model group,WTZR low-dose group,and WTZR high-dose group.The senile osteoporosis(SOP)mouse model was established by D-galactose.Micro-CT scanning analyzed fe-mur microstructure while HE staining detected pathological changes in femur bone tissue samples collected from each experimental con-dition.Furthermore,Western blot was used to detect the senescence-related molecules p21 and p53 and the osteogenesis-related mark-ers OCN and Runx2,qPCR analysis measured tibial expression levels of senescence-related molecules(p21,p53)as well as macro-phage polarisation-related molecules(IL-6,iNOS,CD206,and IL-10)to assess the effect of this compound on a mouse model simula-ting SOP.RESULTS Following intervention with serum containing WTZR,there was a significant decrease in the number of senes-cent positive cells compared to the model group.Additionally,there was a notable decrease in p21 and p53 mRNA and proteins expres-sion(P＜0.05,P＜0.01,P＜0.001).Furthermore,drug-containing WTZR effectively inhibited ROS production induced by H2 O2 and mitigated mitochondrial membrane potential reduction in macrophages(P＜0.05,P＜0.001).Treatment with drug-containing WTZR re-sulted in down-regulated mRNA expression of M1-related gene iNOS(P＜0.05)while up-regulating mRNA expression of M2-related genes CD163 and CD206(P＜0.05).The drug-containing WTZR significantly reduced fluorescence intensity for iNOS(P＜0.01)while increasing ARG1(P＜0.05)fluorescence intensity.Moreover,conditioned medium from macrophages treated with drug-containing ser-um increased ALP positive cell count(P＜0.01,P＜0.001),alizarin red positive area(P＜0.05),as well as Col1a1,Runx2 and OCN mRNA expression levels(P＜0.05,P＜0.01).The Tb.N,BMD,and BV/TV were significantly higher in the WTZR group compared to the model group(P＜0.05,P＜0.01,P＜0.001);meanwhile,Tb.Sp was notably lower than that observed in the model group(P＜0.05,P＜0.01);bone trabeculae were significantly improved,increased in number and widened.Additionally,the compound could significantly inhibit the D-galactose induced up-regulation of tibial senescence-related genes and proteins p21 and p53(P＜0.05,P＜0.001,P＜0.0001),promote the expression of osteogenesis-related markers OCN and Runx2 protein(P＜0.01,P＜0.0001),promote the down-regulation of M1 related genes IL-6 and iNOS(P＜0.05),and promote the expression of M2 related genes IL-10 and CD206(P＜0.05,P＜0.01).CONCLUSION Wenshen Tongluo Zhitong Recipe may play an anti-osteoporosis effect by inhibiting macrophage senescence and promoting the osteogenic differentiation of BMSC.