Nature killer(NK)cells are lymphocytes of the innate immune that play a significant role in aging through direct cell killing and secretion of cytokines.NK cells can delay the occurrence of age-related diseases and prolong lifespan by eliminating senescencent cells.Significant progress has been achieved in the research utilizing NK cells to improve aging.This review aims to summarize that the recent research on the mechanism of NK cells in the context of senescence, as well as the progress made in anti-senescence interventions through animal and clinical studies.Nevertheless, substantial challenges remain for the clinical application of NK cell-based anti-senescence therapies.

Objective To investigate the changes and therapeutic effects of Ginkgo biloba extract(GBE),Donepezil and combination therapy on gray matter volume and cortical morphology in patients with cognitive dysfunction.Methods According to different treatment methods,59 patients with mild cognitive impairment and Alzheimer's disease were divided into GBE group(n=20),Donepezil group(n=20)and combined treatment group(n=19).Before and after treatment,the patients were evaluated by MMSE,Alzheimer's disease assessment scale-cognitive subscale(ADAS-cog),instrumental activities of daily living scale(IADL),geriatric depression scale(GDS),neuropsychiatric inventory(NPI),and quality of life-Alzheimer's disease scale(QOL-AD).Before and after treatment,T1-weighted structural sequence images were acquired using cranial MRI,and voxel-based morphometry(VBM)and cortical morphological analyses were performed.Results There was no significant difference in the scores of MMSE,ADAS-cog,GDS,NPI,IADL and QOL-AD among the GBE group,Donepezil group and combined treatment group before and after treatment(all P>0.05).Compared with those before treatment,there were significant differences in MMSE and ADAS-cog scores in the GBE group,MMSE,ADAS-cog,IADL and NPI scores in Donepezil group,and MMSE,ADAS-cog and IADL scores in combined treatment group after treatment(P＜0.05-0.01).The volume of gray matter in the cerebellar region of GBE group was significantly increased(voxel P＜0.005,cluster P＜0.05,GRF corrected,one-tailed test).In the left putamen region,the difference of gray matter volume in the combined treatment group was significantly higher than that in the GBE group and Donepezil group(all P＜0.01).In the right putamen area,there was no significant difference among the three groups(all P>0.05).In the right hippocampus,the difference in gray matter volume between combined treatment group and GBE group was significantly higher than that in Donepezil group(all P＜0.05).In the left hippocampus,the difference in gray matter volume between GBE group and combined treatment group was significantly higher than that in Donepezil group(all P＜0.001).Correlation analysis showed that the volume of left and right hippocampal gray matter was positively correlated with MMSE before treatment(r=0.352,P=0.008;r=0.424,P=0.001),and negatively correlated with ADAS-cog(r=-0.336,P=0.012;r=-0.362,P=0.007)and IADL scores(r=-0.345,P=0.01;r=-0.312,P=0.02);after treatment,the gray matter volume of the left and right hippocampus was strong positively correlated with MMSE(r=0.582,P＜0.001;r=0.560,P＜0.001),strong negatively correlated with ADAS-cog(r=-0.512,P＜0.001;r=-0.567,P＜0.001)and IADL(r=-0.454,P＜0.001;r=-0.435,P＜0.001).The difference of gray matter volume in the right hippocampus before and after treatment was only weakly negatively correlated with GDS score(r=-0.269,P=0.047),the gray matter volume of right hippocampus was positively correlated with GDS score before treatment(r=0.451,P＜0.001),and this correlation disappeared after intervention(r=0.131,P=0.340).There was no correlation between the difference of gray matter volume before and after treatment in the left hippocampus and the neuropsychological test scores(all P>0.05).The cortex of the superior parietal lobule and inferior parietal lobe in the combined treatment group was significantly thickened(all P＜0.05,FWE corrected).Conclusions GBE,Donepezil and the combined therapy have significant effects on the reduction of cognitive function,and the changes of brain gray matter and cortical morphology are different.GBE improve the volume of the posterior superior cerebellum and other regions,and the combined therapy improve the cortical thickness of the superior parietal lobule and the inferior parietal lobule.

Objective:To investigate the value of T helper 17 cells（Th17）/regulatory T cells（Treg）imbalance in the evaluation of intravenous immunoglobulin（IVIG）resistance in children with Kawasaki disease and Kobayashi score≤4.Methods:A total of 78 children with Kawasaki disease and Kobayashi score ≤ 4 admitted to Hunan Children's Hospital from January 2020 to December 2023 were prospectively selected as the study subjects，all of whom received IVIG treatment.In the acute phase，the proportion of Th17 cells and Treg cells was detected.Children were divided into IVIG sensitive group and IVIG resistance group based on their responsiveness to IVIG treatment.Baseline data of children with different IVIG treatment responsiveness，acute Th17 cell inflammatory factors [interleukin（IL）-17，IL-21，tumor necrosis factor-α（TNF-α）]，Treg cell inflammatory factors [IL-10，IL-35，transforming growth factor-β（TGF-β）] levels，and Th17/Treg values were compared.The correlation between Th17/Treg values and IVIG resistance in children with Kawasaki disease was analyzed using a restricted cubic spline model（RCS）.According to the threshold of correlation between Th17/Treg values obtained from RCS analysis and drug resistance in children，Th17/Treg was grouped，with a focus on analyzing the predictive value and clinical benefits of Th17/Treg values for IVIG resistance in children with Kawasaki disease.Results:Among the 78 children with Kawasaki disease，16 were resistant to IVIG treatment，accounting for 20.51%.The levels of C-reactive protein（CRP）,IL-17,and Th17/Treg in the acute phase of children in the IVIG resistance group were higher than those in the IVIG sensitive group，while the levels of IL-10 were lower than those in the IVIG sensitive group（ P<0.05）.RCS analysis showed that there was a non-linear dose-response relationship between IVIG resistance and acute Th17/Treg values in children with Kawasaki disease（ P<0.05）.When the acute Th17/Treg value was greater than 1.05，the risk of IVIG resistance in children with Kawasaki disease increased with the increase in indicator levels.The levels of CRP and IL-17 in the acute phase of children with Th17/Treg>1.05 were higher than those in the Th17/Treg < 1.05 group，while IL-10 levels were lower than those in the Th17/Treg<1.05 group.The proportion of children resistant to IVIG treatment was higher than that in the Th17/Treg<1.05 group（ P<0.05）.Multivariate Logistic regression analysis showed that CRP，IL-17，IL-10，and Th17/Treg were the influencing factors of IVIG resistance in children with Kawasaki disease（ P<0.05）.It was found through a nomogram that the C-index of the acute phase Th17/Treg values and their secretion of inflammatory factors in children with Kawasaki disease and Kobayashi score ≤ 4，as well as other major indicators，predicted the risk of IVIG resistance.The C-index was 0.975（95% CI 0.944-1.000），indicating good discrimination.When drawing the decision curve，it was found that compared to using each indicator separately，the Th17/Treg value and its secreted inflammatory factors in the acute phase assisted other major indicators in drawing the decision curve with a higher net benefit rate，with a maximum net benefit rate of 0.205. Conclusion:IVIG resistance in children with Kawasaki disease and Kobayashi score≤4 is related to Th17/Treg imbalance.When the Th17/Treg value in the acute phase of the disease is greater than 1.05，the risk of IVIG resistance is higher.The inflammatory factors IL-17 and IL-10 secreted by the two can assist other known indicators related to IVIG resistance in Kawasaki disease patients，improving the accuracy of predicting resistance risk.

Objective To investigate the changes and therapeutic effects of Ginkgo biloba extract(GBE),Donepezil and combination therapy on gray matter volume and cortical morphology in patients with cognitive dysfunction.Methods According to different treatment methods,59 patients with mild cognitive impairment and Alzheimer's disease were divided into GBE group(n=20),Donepezil group(n=20)and combined treatment group(n=19).Before and after treatment,the patients were evaluated by MMSE,Alzheimer's disease assessment scale-cognitive subscale(ADAS-cog),instrumental activities of daily living scale(IADL),geriatric depression scale(GDS),neuropsychiatric inventory(NPI),and quality of life-Alzheimer's disease scale(QOL-AD).Before and after treatment,T1-weighted structural sequence images were acquired using cranial MRI,and voxel-based morphometry(VBM)and cortical morphological analyses were performed.Results There was no significant difference in the scores of MMSE,ADAS-cog,GDS,NPI,IADL and QOL-AD among the GBE group,Donepezil group and combined treatment group before and after treatment(all P>0.05).Compared with those before treatment,there were significant differences in MMSE and ADAS-cog scores in the GBE group,MMSE,ADAS-cog,IADL and NPI scores in Donepezil group,and MMSE,ADAS-cog and IADL scores in combined treatment group after treatment(P＜0.05-0.01).The volume of gray matter in the cerebellar region of GBE group was significantly increased(voxel P＜0.005,cluster P＜0.05,GRF corrected,one-tailed test).In the left putamen region,the difference of gray matter volume in the combined treatment group was significantly higher than that in the GBE group and Donepezil group(all P＜0.01).In the right putamen area,there was no significant difference among the three groups(all P>0.05).In the right hippocampus,the difference in gray matter volume between combined treatment group and GBE group was significantly higher than that in Donepezil group(all P＜0.05).In the left hippocampus,the difference in gray matter volume between GBE group and combined treatment group was significantly higher than that in Donepezil group(all P＜0.001).Correlation analysis showed that the volume of left and right hippocampal gray matter was positively correlated with MMSE before treatment(r=0.352,P=0.008;r=0.424,P=0.001),and negatively correlated with ADAS-cog(r=-0.336,P=0.012;r=-0.362,P=0.007)and IADL scores(r=-0.345,P=0.01;r=-0.312,P=0.02);after treatment,the gray matter volume of the left and right hippocampus was strong positively correlated with MMSE(r=0.582,P＜0.001;r=0.560,P＜0.001),strong negatively correlated with ADAS-cog(r=-0.512,P＜0.001;r=-0.567,P＜0.001)and IADL(r=-0.454,P＜0.001;r=-0.435,P＜0.001).The difference of gray matter volume in the right hippocampus before and after treatment was only weakly negatively correlated with GDS score(r=-0.269,P=0.047),the gray matter volume of right hippocampus was positively correlated with GDS score before treatment(r=0.451,P＜0.001),and this correlation disappeared after intervention(r=0.131,P=0.340).There was no correlation between the difference of gray matter volume before and after treatment in the left hippocampus and the neuropsychological test scores(all P>0.05).The cortex of the superior parietal lobule and inferior parietal lobe in the combined treatment group was significantly thickened(all P＜0.05,FWE corrected).Conclusions GBE,Donepezil and the combined therapy have significant effects on the reduction of cognitive function,and the changes of brain gray matter and cortical morphology are different.GBE improve the volume of the posterior superior cerebellum and other regions,and the combined therapy improve the cortical thickness of the superior parietal lobule and the inferior parietal lobule.

Objective:To investigate the value of T helper 17 cells（Th17）/regulatory T cells（Treg）imbalance in the evaluation of intravenous immunoglobulin（IVIG）resistance in children with Kawasaki disease and Kobayashi score≤4.Methods:A total of 78 children with Kawasaki disease and Kobayashi score ≤ 4 admitted to Hunan Children's Hospital from January 2020 to December 2023 were prospectively selected as the study subjects，all of whom received IVIG treatment.In the acute phase，the proportion of Th17 cells and Treg cells was detected.Children were divided into IVIG sensitive group and IVIG resistance group based on their responsiveness to IVIG treatment.Baseline data of children with different IVIG treatment responsiveness，acute Th17 cell inflammatory factors [interleukin（IL）-17，IL-21，tumor necrosis factor-α（TNF-α）]，Treg cell inflammatory factors [IL-10，IL-35，transforming growth factor-β（TGF-β）] levels，and Th17/Treg values were compared.The correlation between Th17/Treg values and IVIG resistance in children with Kawasaki disease was analyzed using a restricted cubic spline model（RCS）.According to the threshold of correlation between Th17/Treg values obtained from RCS analysis and drug resistance in children，Th17/Treg was grouped，with a focus on analyzing the predictive value and clinical benefits of Th17/Treg values for IVIG resistance in children with Kawasaki disease.Results:Among the 78 children with Kawasaki disease，16 were resistant to IVIG treatment，accounting for 20.51%.The levels of C-reactive protein（CRP）,IL-17,and Th17/Treg in the acute phase of children in the IVIG resistance group were higher than those in the IVIG sensitive group，while the levels of IL-10 were lower than those in the IVIG sensitive group（ P<0.05）.RCS analysis showed that there was a non-linear dose-response relationship between IVIG resistance and acute Th17/Treg values in children with Kawasaki disease（ P<0.05）.When the acute Th17/Treg value was greater than 1.05，the risk of IVIG resistance in children with Kawasaki disease increased with the increase in indicator levels.The levels of CRP and IL-17 in the acute phase of children with Th17/Treg>1.05 were higher than those in the Th17/Treg < 1.05 group，while IL-10 levels were lower than those in the Th17/Treg<1.05 group.The proportion of children resistant to IVIG treatment was higher than that in the Th17/Treg<1.05 group（ P<0.05）.Multivariate Logistic regression analysis showed that CRP，IL-17，IL-10，and Th17/Treg were the influencing factors of IVIG resistance in children with Kawasaki disease（ P<0.05）.It was found through a nomogram that the C-index of the acute phase Th17/Treg values and their secretion of inflammatory factors in children with Kawasaki disease and Kobayashi score ≤ 4，as well as other major indicators，predicted the risk of IVIG resistance.The C-index was 0.975（95% CI 0.944-1.000），indicating good discrimination.When drawing the decision curve，it was found that compared to using each indicator separately，the Th17/Treg value and its secreted inflammatory factors in the acute phase assisted other major indicators in drawing the decision curve with a higher net benefit rate，with a maximum net benefit rate of 0.205. Conclusion:IVIG resistance in children with Kawasaki disease and Kobayashi score≤4 is related to Th17/Treg imbalance.When the Th17/Treg value in the acute phase of the disease is greater than 1.05，the risk of IVIG resistance is higher.The inflammatory factors IL-17 and IL-10 secreted by the two can assist other known indicators related to IVIG resistance in Kawasaki disease patients，improving the accuracy of predicting resistance risk.

Nature killer(NK)cells are lymphocytes of the innate immune that play a significant role in aging through direct cell killing and secretion of cytokines.NK cells can delay the occurrence of age-related diseases and prolong lifespan by eliminating senescencent cells.Significant progress has been achieved in the research utilizing NK cells to improve aging.This review aims to summarize that the recent research on the mechanism of NK cells in the context of senescence, as well as the progress made in anti-senescence interventions through animal and clinical studies.Nevertheless, substantial challenges remain for the clinical application of NK cell-based anti-senescence therapies.

Objective:To detect the expression of microRNA (miR)-211-5p, erythropoietin hepatocyte kinase receptor B2 (EphB2) and erythropoietin hepatocyte kinase ligand B2 (ephrin B2) in spinal cord tissues as well as nerve cells after spinal cord injury (SCI), and to explore their mechanisms and effects on neurological recovery in SCI rats.Methods:The study was conducted from May 2020 to June 2021 using Sprague Dawley (SD) rats and PC12 cells. SD rats were divided into sham-operated group and SCI group of 30 rats each, and Basso-Beattie-Bresnahan (BBB) score were performed at different postoperative time points (1, 3, 7, 14, 21 and 28 d), and the relative expression of miR-211-5p and Eph/ephrin B2 mRNA was measured by quantitative real-time polymerase chain reaction (qPCR); the SCI rats were divided into recombinant lentiviral vector LV-miR-211-5p group (group A), empty lentiviral vector LV-eGFP (group B) and saline group (group C), with 15 rats in each group, respectively. The recombinant lentiviral vector, empty lentiviral vector and saline were injected on the cephalic and caudal sides of the spinal cord injury, and the relative expression of miR-211-5p and Eph/ephrin B2 mRNA in the spinal cord tissue was measured at 1, 7 and 14 d after surgery. In addition, a PC12 injury cell line model was established with 150 μmol/L hydrogen peroxide (H 2O 2), and the apoptosis rate and apoptosis-related proteins and contents of different cell lines were detected by flow cytometry and Western blot, respectively. MiR-211-5p was verified to target EphB2 by dual luciferase reporter gene. Results:The results of the animal experiments showed that at different postoperative time points, the miR-211-5p levels in the SCI group were lower than those in the SHAM group: 0.70 ± 0.03 vs. 1.00 ± 0.10, 0.60 ± 0.04 vs. 1.00 ± 0.05, 0.45 ± 0.10 vs. 1.00 ± 0.12, 0.30 ± 0.06 vs. 1.00 ± 0.15, 0.20 ± 0.05 vs. 1.00 ± 0.13, 0.10 ± 0.02 vs. 1.00 ± 0.07. In contrast, levels of Eph/ephrin B2 were higher in the SCI group compared to the SHAM group: 1.10 ± 0.05 vs. 1.00 ± 0.01, 1.80 ± 0.01 vs. 1.00 ± 0.08, 2.30 ± 0.01 vs. 1.00 ± 0.10, 2.60 ± 0.01 vs. 1.00 ± 0.05, 2.80 ± 0.01 vs. 1.00 ± 0.06, 3.00 ± 0.01 vs. 1.00 ± 0.07 and 1.20 ± 0.05 vs. 1.00 ± 0.02, 1.60 ± 0.01 vs. 1.00 ± 0.03, 2.10 ± 0.10 vs. 1.00 ± 0.01, 2.40 ± 0.11 vs. 1.00 ± 0.09, 2.70 ± 0.13 vs. 1.00 ± 0.05, 2.90 ± 0.12 vs. 1.00 ± 0.03 ( P<0.05). At 14 d after surgery, Group A exhibited higher BBB scores than Groups B and C: (14.0 ± 1.1) points vs. (8.0 ± 1.1) and (8.2 ± 1.2) points, while miR-211-5p levels were higher than those in Groups B and C: 1.90 ± 0.10 vs. 0.40 ± 0.01 and 0.50 ± 0.02, and Eph/ephrin B2 levels were lower than those in Groups B and C: 0.70 ± 0.10 vs. 1.80 ± 0.04 and 1.90 ± 0.06, 0.60 ± 0.03 vs. 2.00 ± 0.04 and 2.10 ± 0.05 ( P<0.05). Immunofluorescence staining showed that the levels of GAP-43 and synaptophysin in group A were higher than those in groups B and C at 14 d after surgery ( P<0.05). Cellular assays showed that overexpression of miR-211-5p inhibited the apoptosis rate of H 2O 2-induced PC12 cells and the expression of the apoptosis-related gene Cleaved-caspase3 ( P<0.05). Knockdown of miR-211-5p increased the apoptosis rate of H 2O 2-induced PC12 cells and the expression of the apoptosis-related gene Cleaved-caspase3 ( P<0.05). Dual luciferase reporter gene assay confirmed that EphB2 was a target gene of miR-211-5p and overexpression of EphB2 antagonized the inhibitory apoptosis effect of miR-211-5p on H 2O 2-induced PC12 cells. Conclusions:This study showed that miR-211-5p could promote neurological repair in SCI by inhibiting the expression of Eph/ephrin B2 signaling pathway, suggesting that using miR-211-5p as a target to inhibit Eph/ephrin B2 signaling pathway may have a protective effect on SCI.

Objective To screen differentially expressed miRNAs(DEMs)by comparing the expression of miRNAs in serum exosomes between Alzheimer's disease(AD)patients and healthy controls.Methods A total of 71 AD patients admitted to Department of Geriatric Neurology of Xiangya Hospital from March 2017 to August 2018 and another 71 healthy individuals who taking physical examination in the hospital during same period were recruited and assigned into AD and HC groups,respectively.Four AD patients and four healthy subjects were selected for high-throughput second-generation sequencing of exosome miRNAs.The results were analyzed to obtain the DEMs between them,and the top 4 DEMs were finally selected.Then real-time quantitative real-time PCR was applied for all the subjects to detect the expression of the 4 DEMs.Results High-throughput second-generation sequencing detected 775 miRNAs,and 44 DEMs were found with statistical difference between the 2 groups(P＜0.05).Compared with the HC group,34 miRNAs were up-regulated and 10 were down-regulated in the AD group.The top 4 DEMs were miRNA-148a-3p,miRNA-16-5p,miRNA-19b-3p and miRNA-483-5p.MiRNA-148a-3p was significantly up-regulated in the AD group than the HC group(P＜0.01),but there were no significant differences in the expression level in the other 3 DEMs between the 2 groups.ROC curve analysis showed that the area under the curve of miRNA-148a-3p was 0.7113(95％CI:0.622～0.801),with a sensitivity of 71.6％and a specificity of 69.7％.Conclusion Serum exosome miRNA-148a-3p can be used as a biomarker for the diagnosis of AD.

Objective To explore the compensatory effect of exercise on brain activation abnormalities caused by sleep disorders.Methods Brain imaging meta-analysis was conducted using activation likelihood estimation(ALE)to investigate brain activation abnormalities caused by sleep disorders and changes in brain activation related to exercise.Results A total of 119 studies were included,and it was found that there was widespread activation in the bilateral frontal areas,hippocampus,superior and middle temporal gyri,and some regions of the thalamus in abnormal brain regions related to sleep disorders and brain regions after exercise intervention.Conclusion Exercise may play an important role in alleviating brain activation abnormalities caused by sleep disorders.

Objective:Evaluation of the clinical value of the BioPerfectus multiplex real time (BMRT)-HPV for cervical cancer screening.Methods:Physician-collected specimens of 1 495 women who were positive of Cobas 4800 HPV (Cobas-HPV), HPV genotyping based on SEQ uencing (SEQ-HPV), and (or) cytology ≥low grade squamous intraepithelial lesion (LSIL) in the primary screening of Chinese Multiple-center Screening Trial (CHIMUST), and 2 990 women selected from those who were negative of primary screening in the same project through nested control randomization with age-matching were tested for BMRT-HPV, which reported type-specific viral loads/10 000 cells in each specimen. With comparing to Cobas-HPV results and taking cervical histopathological diagnosis as the endpoint, the concordance of high-risk (HR)-HPV subtypes among the three assays was explored ,and the sensitivity and specificity of BMRT-HPV for cervical cancer screening were evaluated.Results:(1) The overall agreenment of HR-HPV subtypes between BMRT-HPV and Cobas-HPV, or SEQ-HPV test sample was 94.8%, 94.4%, with Kappa values 0.827, 0.814. (2) The sensitivity and specificity for cervical intraepithelial neoplasia (CIN) Ⅱ + of BMRT-HPV, Cobas-HPV and SEQ-HPV were 92.62%, 94.26%, 93.44% and 84.67%, 83.25%, 82.76%, respectively. There were no significant difference in sensitivity among the three HPV assays (all P>0.05), but the specificity of BMRT-HPV for CIN Ⅱ + was higher than those of Cobas-HPV and SEQ-HPV ( P<0.01). The sensitivity for CIN Ⅲ + of three HPV assays were all 100.00%, and the specificity for CIN Ⅲ + of BMRT-HPV was higher than those of Cobas-HPV and SEQ-HPV (83.40% vs 81.95%, 83.40% vs 81.50%; P<0.01). The number of pathological examinations of colposcopy for cervical biopsy detected in 1 case of CIN Ⅱ + or CIN Ⅲ + in BMRT-HPV was less than those in Cobas-HPV and SEQ-HPV ( P<0.01). When using HPV 16/18 + cytology ≥atypical squamous cell of undetermined signification (ASCUS) to triage HPV positive women among three assays, there was no different in the sensitivities of detecting CIN Ⅱ + and CIN Ⅲ + ( P>0.05). The specificity BMRT-HPV was slightly higher than those in Cobas-HPV or SEQ-HPV (all P<0.05), and the colposcopy referral rate was lower than those in Cobas-HPV and SEQ-HPV (all P<0.05). Conclusions:BMRT-HPV is as sensitive as Cobas-HPV or SEQ-HPV for primary cervical cancer screening, and has higher specificity. Therefore it could be used as a primary screening method for cervical cancer, which is worthy of clinical application.