Objective:To develop a self-locked DNAzyme nanoprobe-based fluorescence amplification strategy for wash-free and ultrasensitive detection of breast cancer-derived small extracellular vesicles (sEV).Method:A DNAzyme self-locked probe was designed to recognize the epithelial cell adhesion molecule (EpCAM) specifically expressed on breast cancer-derived sEVs. Upon binding to EpCAM, the DNAzyme-lock structure was opened, restoring the DNAzyme cleavage activity. The activated DNAzyme then cyclically cleaved the RNA site on the substrate strand. Fluorescently labeled substrate strands were used to detect sEVs at varying concentrations, and the detection limit and linear range were determined.Results:The DNAzyme self-locked probe successfully identified breast cancer-derived sEVs and generated a fluorescent signal through cyclic cleavage. The proposed method achieved wash-free detection of sEVs, with the fluorescence intensity showing a strong linear correlation with sEV concentration ( R2=0.98). The linear detection range was 1.0×10 2-1.0×1.0 7 particles/μl, with a detection limit of 59 particles/μl. Conclusion:This study established a wash-free and highly sensitive strategy for quantifying breast cancer-derived sEVs, which provides a promising technical approach for the early diagnosis of cancer.

OBJECTIVE To explore the effect of Astragali Radix-Curcuma Zedoaria-Paridis Rhizoma(Qi-Zhu-Zao)combina-tion on inhibiting the growth and metastasis of colon cancer based on the PINK1/Parkin/EMT signaling pathway.METHODS Thirty male BALB/c mice were randomly assigned to five groups:sham operation group,model group,positive control group,high-dose Qi-Zhu-Zao group(5.85 g·kg-1),and low-dose Qi-Zhu-Zao group(2.925 g·kg-1),with six mice in each group.An orthotopic colon cancer model was established in the mice using CT26.WT cells.After 15 days of treatment,tumor and liver tissues were collected from each group.Hematoxylin and eosin(HE)staining was performed to assess tumor metastasis,and transmission electron microscopy was used to observe mitochondrial autophagy in tumor tissues.The expression of mitochondrial autophagy-related proteins PINK1,Parkin,p62,and LC3-Ⅱ/LC3-Ⅰ was analyzed using Western blot and immunohistochemistry(IHC).Additionally,the expression levels of epithelial-mesenchymal transition(EMT)-related proteins and mRNA,including E-cadherin,N-cadherin,Vimentin,and Snail,were detected using Western blot,qPCR,and IHC staining.RESULTS Compared to the model group,mice in the treatment groups exhibited significantly reduced tumor volumes and fewer metastatic foci.Additionally,liver tissues showed pathological changes,and the overall growth condition of the mice was markedly improved;the tumor tissues in the treatment groups displayed selective mitochon-drial autophagy,accompanied by the formation of autophagosomes.The treatment influenced the PINK1/Parkin pathway-mediated mi-tochondrial autophagy biological process,with PINK1,Parkin,p62,and LC3-Ⅱ/LC3-Ⅰ levels being significantly upregulated(P<0.05,P<0.01),the high-dose group exhibited a more significant impact than the low-dose group(P<0.05,P<0.01).Furthermore,the treatment groups also showed significant reductions in the protein and mRNA levels of N-cadherin,Vimentin,and Snail(P<0.05,P<0.01),along with significant increases in the protein and mRNA levels of E-cadherin(P<0.05,P<0.01),these effects were more pronounced in the high-dose group compared to the low-dose group(P<0.05,P<0.01).CONCLUSION The herbal combination of Qi-Zhu-Zao inhibits tumor growth and metastasis to a certain extent in a mouse model of orthotopic transplantation of colon cancer.The underlying mechanism may involve the restoration of mitochondrial function through the PINK1/Parkin signaling pathway and the inhibition of the epithelial-mesenchymal transition(EMT)process,thereby achieving a therapeutic effect on colon cancer.

OBJECTIVE To explore the effect of Astragali Radix-Curcuma Zedoaria-Paridis Rhizoma(Qi-Zhu-Zao)combina-tion on inhibiting the growth and metastasis of colon cancer based on the PINK1/Parkin/EMT signaling pathway.METHODS Thirty male BALB/c mice were randomly assigned to five groups:sham operation group,model group,positive control group,high-dose Qi-Zhu-Zao group(5.85 g·kg-1),and low-dose Qi-Zhu-Zao group(2.925 g·kg-1),with six mice in each group.An orthotopic colon cancer model was established in the mice using CT26.WT cells.After 15 days of treatment,tumor and liver tissues were collected from each group.Hematoxylin and eosin(HE)staining was performed to assess tumor metastasis,and transmission electron microscopy was used to observe mitochondrial autophagy in tumor tissues.The expression of mitochondrial autophagy-related proteins PINK1,Parkin,p62,and LC3-Ⅱ/LC3-Ⅰ was analyzed using Western blot and immunohistochemistry(IHC).Additionally,the expression levels of epithelial-mesenchymal transition(EMT)-related proteins and mRNA,including E-cadherin,N-cadherin,Vimentin,and Snail,were detected using Western blot,qPCR,and IHC staining.RESULTS Compared to the model group,mice in the treatment groups exhibited significantly reduced tumor volumes and fewer metastatic foci.Additionally,liver tissues showed pathological changes,and the overall growth condition of the mice was markedly improved;the tumor tissues in the treatment groups displayed selective mitochon-drial autophagy,accompanied by the formation of autophagosomes.The treatment influenced the PINK1/Parkin pathway-mediated mi-tochondrial autophagy biological process,with PINK1,Parkin,p62,and LC3-Ⅱ/LC3-Ⅰ levels being significantly upregulated(P<0.05,P<0.01),the high-dose group exhibited a more significant impact than the low-dose group(P<0.05,P<0.01).Furthermore,the treatment groups also showed significant reductions in the protein and mRNA levels of N-cadherin,Vimentin,and Snail(P<0.05,P<0.01),along with significant increases in the protein and mRNA levels of E-cadherin(P<0.05,P<0.01),these effects were more pronounced in the high-dose group compared to the low-dose group(P<0.05,P<0.01).CONCLUSION The herbal combination of Qi-Zhu-Zao inhibits tumor growth and metastasis to a certain extent in a mouse model of orthotopic transplantation of colon cancer.The underlying mechanism may involve the restoration of mitochondrial function through the PINK1/Parkin signaling pathway and the inhibition of the epithelial-mesenchymal transition(EMT)process,thereby achieving a therapeutic effect on colon cancer.

Objective:To develop a self-locked DNAzyme nanoprobe-based fluorescence amplification strategy for wash-free and ultrasensitive detection of breast cancer-derived small extracellular vesicles (sEV).Method:A DNAzyme self-locked probe was designed to recognize the epithelial cell adhesion molecule (EpCAM) specifically expressed on breast cancer-derived sEVs. Upon binding to EpCAM, the DNAzyme-lock structure was opened, restoring the DNAzyme cleavage activity. The activated DNAzyme then cyclically cleaved the RNA site on the substrate strand. Fluorescently labeled substrate strands were used to detect sEVs at varying concentrations, and the detection limit and linear range were determined.Results:The DNAzyme self-locked probe successfully identified breast cancer-derived sEVs and generated a fluorescent signal through cyclic cleavage. The proposed method achieved wash-free detection of sEVs, with the fluorescence intensity showing a strong linear correlation with sEV concentration ( R2=0.98). The linear detection range was 1.0×10 2-1.0×1.0 7 particles/μl, with a detection limit of 59 particles/μl. Conclusion:This study established a wash-free and highly sensitive strategy for quantifying breast cancer-derived sEVs, which provides a promising technical approach for the early diagnosis of cancer.

Objective:To investigate personal exposures to nitrogen oxides(NOX)and nitrogen di-oxide(NO2)and the influence of baseline personal characteristics,living environment and daily activity patterns of the participants on the exposures among adults over 35 in Tianjin and Shanghai.Methods:In this panel study,91 healthy nonsmoking adults aged over 35 from Tianjin and Shanghai participated in our study.The study was conducted in summer and winter.The participants were followed for three times with an interval of at least two weeks.Only participants in Shanghai were followed once in winter because of the COVID-19 pandemic.Twenty-seven participants completed follow-up visits in both seasons.We measured their 24 h personal exposures to NOX and NO2and collected their baseline and time-activity in-formation through questionnaire/diary.The linear mixed model was used to analyze the associations be-tween potential influencing factors and personal NOX and NO2 exposure levels.Results:There were 349 follow-up visits with valid 24 h personal NO2 and NOX exposure measurements in the two cities.The ave-rage 24 h personal exposures to NO2 and NOX(volume fraction)in Tianjin participants were 18.0 x 10-9 and 26.2 × 10-9 in summer,and 31.0 x 10-9 and 54.9 x 10-9in winter,respectively;and the average 24 h personal exposures to NO2 and NOX in Shanghai participants were 38.7 x 10-9and 100.0x10-9in summer,and 45.5 x10-9 and 139.2 x 10-9 in winter,respectively.The results of univariate regression analysis showed that their personal NOX exposure levels were significantly associated with city,season,gender,average daily cooking times,and ambient NO2 concentrations measured at fixed-site monitoring stations.In addition to the above factors,the personal NOX exposure levels were also significantly associ-ated with educational level and the personal NO2 exposure levels were also significantly associated with passive smoking,average daily home time,cooking energy type,residential distance from main traffic road,and use of kitchen ventilators.Multivariate regression analysis showed that the personal exposure levels of NO2 and NOX were significantly lower in Tianjin than that in Shanghai,were significantly lower in summer than that in winter,and were significantly and positively associated with ambient NO2 concen-trations measured at fixed-site monitoring stations.In addition,personal NOX exposure levels were signifi-cantly lower in females than in males,and personal NO2 exposure levels were significantly positively asso-ciated with average daily cooking times and significantly inversely associated with average daily home time.For every interquartile range(IQR)increase(12.7 × 10-9)in ambient NO2,the personal NO2 exposure levels increased by 27.5％(95％CI:17.0％-38.9％),and personal NOX exposure levels in-creased by 16.1％(95％CI:7.1％-25.8％).Conclusion:Season,city and ambient NO2 concentra-tions are significant influencing factors of personal exposure levels of NO2and NOX At the same time,the personal exposures levels of NO2 are also affected by lifestyle factors.Our study provides scientific evi-dence for making precise air pollution control decisions and reducing the exposure levels of NOX in the population.

Background Regional differences in economic development, natural environment, health care level, and social structure may lead to differences in the provincial distribution of the health status of the elderly population. Objective To explore the provincial distribution characteristics, regional differences, and influencing factors of the self-assessed health of the elderly population, with the aim of providing a policy basis for improving the health of the elderly population and promoting healthy aging according to local conditions. Methods Using 31 provinces (municipalities and autonomous regions) in China as the basicstudy unit and based on the method of Wagstaff, the self-rated health data of the elderly population (aged 60 years and above) in each province from the 2010 and 2020 national censuses and the 2015 1% National Population Sample Survey were converted into ill-health scores as a measure of self-assessed health, and higher scores represented worse health status perception. Global Moran's I was used to evaluate spatial autocorrelation, range [−1, 1], with a value of 1 as a perfect clustered pattern. Local Moran's I was used to evaluate the tendency of local autocorrelation, and high-high aggregation/low-low aggregation indicated that both target province and its neighboring provinces showed higher/lower ill-health scores. Spatial econometric models were selected by Lagrange multiplier test and Hausman test to explore influencing factors of the self-assessed health of the elderly population. Results In 2010, 2015, and 2020, the national ill-health scores of the elderly population were 1.831, 1.873, and 1.547, respectively, and the corresponding Global Moran's I statistics were 0.347, 0.482, and 0.511, respectively (P<0.01), indicating that the ill-health scores of the elderly population showed a significant spatial positive autocorrelation, and the degree of spatial aggregation was increasing gradually. From 2010 to 2020, the high-high aggregation of ill-health scores among the elderly population was concentrated in the inland northwest, while the low-low aggregation was concentrated in the southeast coast, gradually showing a "southeast-central-northwest" stepped incremental pattern of differentiation. The Lagrange multiplier test and Hausman test suggested that the fixed-effects spatial lagged model was a better choice, and the regression model showed a spatial autocorrelation in the ill-health scores of the elderly population, with an autocorrelation coefficient of 0.3969 (P<0.001); the ill-health scores of the elderly population were negatively correlated with the natural logarithms of gross regional product per capita, and the number of beds in health care facilities per 1000 population, with regression coefficients of −0.8297 and −0.0454 (P<0.05) respectively, and positively correlated with the annual average concentration of PM_2.5, illiteracy rate, and the number of health technicians per 1000 population, with regression coefficients of 0.0033, 0.0297, and 0.0765 (P<0.05), respectively. Conclusion From 2010 to 2020, the overall self-assessed health level of China's elderly population showed an upward trend and a spatial positive autocorrelation, with better self-assessed health in the southeast coast and poorer ratings in the northwestern inland. Additionally, there was a gradual decline in self-assessed health of the elderly population from the southeast to the central regions and further to the northwest in terms of spatial distribution. Economic development level, environmental pollution, health resource allocation, and education level are important factors influencing the self-assessed health of the elderly population.

Objective: To analyze the ear, nose, and throat exam of some freshmen in the military college entrance examination in Shandong Province in 2020 and to facilitate adolescent targeted health promotion. Methods: The 1 411 freshmen participating in the military college entrance examination in Jinan, Zibo and Weifang of Shandong Province were included. The ear, nose, and throat exam were performed by professionals using electric otoscope, 5 meter whispering test, and front rhinoscope. Results: Nasal septal deviation and hypertrophy of inferior turbinate accounted for the highest proportion. Among 489 cases of nasal septum deviation, the detection rate of Jinan (15.97%) was significantly lower than that of Weifang (43.60%) and Zibo (46.53%) ( χ 2=63.32， P <0.05). For deviation of nasal septum, the detection rate in students with urban residence (31.53%) was significantly lower than that of rural students (39.03%) ( χ 2=4.11， P <0.05). Seventy two cases of inferior turbinate hyperplasia were detected, and the detection rate in Jinan (2.99%) was significantly lower than that in Weifang (6.51%) and Zibo (6.04%) ( χ 2=6.63， P <0.05). The detection rate of tonsil hypertrophy was significantly lower in boys (4.63%), students from urban area (3.94%), compared with that of girls(9.56%) and rural students (6.70%) ( χ 2=5.35，4.86， P <0.05). In pharyngeal examination, tonsil hyperplasia was the most common condition of enlarged tonsils ( n =214), which was significantly higher in Jinan(22.36%) than that of Weifang (11.71 %) and Zibo (10.74%) ( χ 2=22.39， P <0.05), and was significantly lower in boys (14.38%) and rural students (12.40%) than that in girls (22.79%) and urban students (17.24%) ( χ 2=4.70，4.65， P <0.05). Conclusion Nasal septum deviation and tonsil hypertrophy are the most prevalent upper airway diseases among freshmen participating in the military college entrance examination. Prevention and treatment of nasopharynx diseases should be emphasized.

BackgroundDue to the COVID-19 pandemic， both teenagers' studies and personal life are critically affected， which has resulted in a variety of mental health problems. In this regard， scholars at home and abroad have carried out a large number of research concerning adolescent mental health， of which there still exists a lack of systematic combing and review. ObjectiveTo understand the status and development trend of research on adolescent mental health during the COVID-19 pandemic at home and abroad， and to grasp the current research hotspots and trends in this field， so as to provide references for relevant research and practice in the post-epidemic era. MethodsOn October 30， 2022， we searched through China Knowledge Network Infrastructure （CNKI） and Web of Science database， and the publishing time of articles to be retrieved was limited between December 1， 2019 and October 30， 2022. Excel and CiteSpace were used to perform visual analysis on these articles in terms of number， author， institution， country and keywords of the articles. ResultsA total of 7 608 articles were included. At home and abroad， the number of papers related to adolescent mental health generally increased at first and then decreased under the pandemic situation. Compared with foreign countries， the connection and cooperation among domestic scholars and institutions was not close enough. The top three countries in the number of English literature published were the United States， Britain and China， and those in intermediary center were Tunis， Cameroon and Anguilla. The parent-child relationship and mental health of teenagers during were much concerned by scholars both at home and abroad. With the passage of time， researchers at home and abroad had shifted their focus from only negative factors to positive factors. ConclusionChinese scholars or institutions need to strengthen more domestic and international exchanges and cooperation. Scholars from different countries can carry out cross-cultural study on research topics of common concern， and continue to explore the positive psychological changes of teenagers in the post-epidemic era.［Funded by National Social Science Foundation 2020 Education Youth Project of 13th Five-Year Plan （number， CHA200259）］

Objective:To compare the pharmacokinetics and safety of single intravenous injection of the generic bevacizumab injection WBP264 and the original bevacizumab injection Avastin ? in healthy male volunteers. Methods:The study was designed as a randomized, double-blind, single dose, parallel, and controlled phase I clinical trial. Healthy male volunteers who were recruited publicly were randomized into the trial group (intravenous infusion of WBP264) and the control group (intravenous infusion of Avastin ?), and the dose was 3 mg/kg. Peripheral venous blood was collected within 30 minutes before administration, 45 minutes after onset of the administration, immediately after finishing the administration, 2.5, 3.5, 5.5, 9.5, 13.5, 24, 48 hours and on the 5th, 8th, 15th, 22nd, 29th, 36th, 43rd, 57th, 71st, 85th, and 99th days after the administration. The plasma concentration was measured by enzyme-linked immunosorbent assay, the plasma concentration-time curve and its semilogarithmic plot were plotted, and the pharmacokinetic parameters such as the area under the plasma concentration-time curve [including AUC from time zero to the time of the last quantifiable concentration (AUC 0-t) and AUC from time zero to infinity (AUC 0-∞)], peak concentration ( Cmax), time to peak ( Tmax), plasma elimination half-life ( t1/2), clearance rate (CL), and apparent volume of distribution (Vd) were calculated. When the 90% confidence intervals ( CI) of the geometric mean ratio of AUC 0-t, AUC 0-∞, and Cmax between the trial group and the control group were between 0.80-1.25, it indicated that pharmacokinetics of WBP264 and Avastin ? were similar. The physical examination, vital signs detection, electrocardiogram, and laboratory tests were performed on the subjects, the occurrence of adverse events (AEs) and the severity classification were recorded, and correlation between the AEs and the trial drug was evaluated. The anti-drug antibody and its neutralizing antibody were detected before administration and on the 8th, 15th, 29th, 43rd, 71st, and 99th days after administration to evaluate the immunogenicity of the drug. Results:A total of 78 subjects were recruited, 39 in the trial group and 39 in the control group. In the trial group, 2 cases withdrew from the trial (1 case did not take the drug and 1 case withdrew for personal reason after taking the drug). Seventy-seven cases were in the safety analysis set and 76 cases in the pharmacokinetic analysis set. The differences in age, height, weight, and body mass index between the 2 groups were not significant (all P>0.05). The plasma concentration-time curves of bevacizumab between the trial group and the control group were similar. The geometric mean ratios (90% CI) of AUC 0-t, AUC 0-∞, and Cmax were 1.04 (0.98-1.10), 1.03 (0.98-1.10), and 1.09 (1.03-1.14), respectively. The differences in the incidence of overall AEs [89.5% (34/38) vs. 87.2% (34/39)] and the incidence of AEs possibly related to the trial drug [86.8% (33/38) vs. 79.5% (31/39)] between the trial group and the control group were not significant (all P>0.05). Only one case of AE in the trial group was grade 3 in severity and was assessed as being not related to the drug, and the rest were grade 1-2, with grade 1 AEs accounting for the vast majority. The difference in the positive rate of anti-drug antibody between the trial group and the control group was not significant [10.5% (4/38) vs. 10.3% (4/39), P>0.05]. The neutralizing antibody test was negative in the patients with positive anti-drug antibody. Conclusion:The pharmacokinetics and safety of WBP264 and Avastin ? are similar.

Objective: To investigate the reversal of drug resistance of drug-resistant hepatoma cell line HepG2/DDP by emetine and its mechanism. Methods : The drug-resistant hepatoma cell line HepG2/DDP was established by high-dose cisplatin(DDP) shock combined with low-dose continuous induction. Reverse virtual screening, molecular docking, surface plasmon resonance(SPR) and Western blot assays were employed to explore the target of emetine. The target was validated by transfection assayin vivo. The inhibitory effect of emetine combined with DDP on the proliferation of HepG2/DDP cells was detected by cytotoxicity assay. The sensitization effect of emetine was verified by flow cytometry, and the expression of the apoptosis related proteins BCL2, Bax and Cleaved-caspase-3 were analyzed by Western blot. Results : Emetine enhanced the sensitivity of HepG2/DDP cells to DDP and reduced the resistance index(RI) from 3.69 to 0.93. Reverse virtual screening, molecule docking and SPR results showed that emetine can stably bind to PARP-1. Western blot assays showed that emetine had a potent enzymatic inhibitory activity against PARP-1in vivo. Furthermore, emetine's potentiation of DDP in HEPG2/DDP cells nearly disappeared when the cells were transfected with siRNAs against PARP-1. Flow cytometry showed that emetine could enhance the proapoptotic effect of DDP on HepG2/DDP cells. Conclusion Emetine can enhance the sensitivity of HepG2/DDP cells to DDP, and its mechanism may be related to its inhibition of PARP-1.