The study aimed to construct the second and third generation chimeric antigen receptor T cells (CAR-T) targeting the c-mesenchymal-epithelial transition factor (c-Met) protein, and observe their killing effect on human serous ovarian cancer cell SKOV-3. The expression of MET gene in ovarian serous cystadenocarcinoma, the correlation between MET gene expression and the abundance of immune cell infiltration, and the effect of MET gene expression on the tissue function of ovarian serous cystadenocarcinoma were analyzed by bioinformatics. The expression of c-Met in ovarian cancer tissues and adjacent tissues was detected by immunohistochemical staining. The second and third generation c-Met CAR-T cells, namely c-Met CAR-T(2G/3G), were prepared by lentivirus infection, and the cell subsets and infection efficiency were detected by flow cytometry. Using CD19 CAR-T and activated T cells as control groups and A2780 cells with c-Met negative expression as Non target groups, the kill efficiency on SKOV-3 cells with c-Met positive expression, cytokine release and cell proliferation of c-Met CAR-T(2G/3G) were explored by lactate dehydrogenase (LDH) release, ELISA and CCK-8 respectively. The results showed that MET gene expression was significantly up-regulated in ovarian cancer tissues compared with normal tissues, which was consistent with the immunohistochemistry results. However, in all pathological stages, there was no obvious difference in MET expression and no correlation between MET gene expression and the race and age of ovarian cancer patients. The second generation and third generation c-Met CAR-T cells were successfully constructed. After lentivirus infection, the proportion of CD8⁺ T cells in c-Met CAR-T(2G) was upregulated, while there was no significant change in the cell subsets of c-Met CAR-T(3G). The LDH release experiment showed that the kill efficiency of c-Met CAR-T(2G/3G) on SKOV-3 increased with the increase of effect-target ratio. When the effect-target ratio was 20:1, the kill efficiency of c-Met CAR-T(2G) reached (42.02 ± 5.17)% (P < 0.05), and the kill efficiency of c-Met CAR-T(3G) reached (51.40 ± 2.71)% (P < 0.05). ELISA results showed that c-Met CAR-T released more cytokine compared to CD19 CAR-T and activated T cells (P < 0.05). Moreover, the cytokine release of c-Met CAR-T(3G) was higher than c-Met CAR-T(2G) (P < 0.01). The CCK-8 results showed that after 48 h, the cell number of c-Met CAR-T(2G) was higher than that of c-Met CAR-T(3G) (P < 0.01). In conclusion, both the second and third generation c-Met CAR-T can target and kill c-Met-positive SKOV-3 cells, with no significant difference. c-Met CAR-T(2G) has stronger proliferative ability, and c-Met CAR-T(3G) releases more cytokines.
Humans ; Female ; Ovarian Neoplasms/immunology* ; Proto-Oncogene Proteins c-met/metabolism* ; Receptors, Chimeric Antigen/immunology* ; Cell Line, Tumor ; Cystadenocarcinoma, Serous/immunology* ; T-Lymphocytes/immunology*

In modern society, sugary foods have become an integral part of many people′s lives. However, excessive sugar consumption has adverse effects on both overall health and oral health, serving as a contributing factor to the global increasing incidence in oral diseases, cardiovascular diseases, cancers, obesity, and diabetes. In response to the health risks related to high-sugar diets, the World Health Organization (WHO) and World Dental Federation (FDI) have proposed initiatives and recommendations, with various governments implementing different policies and strategies to reduce sugar intake. Chinese government has also taken proactive measures. The "Healthy China Action (2019-2030)" initiative introduced by the State Council in 2019 established a crucial benchmark in limiting the average daily intake of added sugar to 25 g per person forward to 2030. Experts from Chinese Center for Disease Control and Prevention and the field of oral health have meticulously examined the impacts of sugar reduction on oral health, as well as strategies, methods, and practical considerations related to reducing sugar intake through several meeting and wrote the "Expert consensus: reducing free-sugar for caries prevention", which was subsequently reviewed and revised based on the feedback from multiple stakeholders. They have conducted thorough analyses of global trends in sugar reduction and best practices to provide valuable insights to China for crafting effective policies and strategies on sugar reduction. This consensus mainly includes the classification of free sugars, the latest scientific evidence on dental caries, recommendations from WHO on sugar-sweetened beverage taxes, nutrition labeling, advertising, food reform, adjusting supply systems, education, and promotion strategies, as well as sugar reduction actions taken by various governments around the world. Combining the actual situation in China, policy recommendations and authoritative popular science knowledge on sugar reduction for caries prevention to public are proposed to advocate for experts in multiple fields to focus on sugar reduction for caries prevention, promote the work process, and provide the scientific basis for oral health educators.

Objective:To construct a questionnaire to measure parents′ behavioral and social drivers (BeSD) in administering rotavirus vaccine to their children, and to test the reliability and validity of the questionnaire.Methods:A questionnaire was constructed to investigate the BeSD of rotavirus vaccination among parents based on the WHO BeSD questionnaire. A survey was conducted among parents of children aged 0-3 years old in vaccination clinics from 15 community health service centers in Nanjing City of Jiangsu Province, Yuhuan and Kaihua Counties of Zhejiang Province, Anqing City of Anhui Province, and Yangpu District of Shanghai City from April to May in 2024. Exploratory factor analysis (EFA) and confirmatory factor analysis (CFA) were used to analyze the reliability (internal consistency reliability, combination reliability and split half reliability) and validity (structural validity, convergence validity and discriminative validity) of the questionnaire.Results:A total of 850 questionnaires were collected, of which 614 were valid, with an effective response rate of 72.20%. The EFA resulted in the refinement of the rotavirus vaccine BeSD questionnaire from 37 to 35 items (including 24 core items), and suggested a five-factor structure including the perceptions of vaccine, confidence in vaccine, social process of vaccination, motivation for vaccination, and practical problems of vaccination. The cumulative variance contribution rate reached 56.27%. The CFA confirmed an ideal five-factor model fit (GFI=0.82,CFI=0.86,PNFI=0.67,RMSEA=0.07). The AVE of each dimension was greater than 0.50. The AVE square root of each dimension of the questionnaire was greater than its correlation coefficient. The Cronbach′s α coefficient of the total questionnaire was 0.79.Conclusion:The developed BeSD questionnaire of rotavirus vaccine has good reliability and validity, and can be used to measure parents′ behavioral and social factors in administering rotavirus vaccine to their children.

[Purpose]To analyze the incidence and mortality of thyroid cancer in Zhejiang cancer registration areas in 2021 and trends from 2000 to 2021.[Methods]Based on cancer data from 22 registries across Zhejiang Province between 2000 and 2021,the crude incidence/mortality rates,age-standardized incidence/mortality rates of thyroid cancer by Chinese and world standard population(ASIRC/ASMRC,ASIRW/ASMRW),cumulative rate(0～74 years old)and truncated rate(35～64 years old)were analyzed.Joinpoint regression model was employed to calculate the annual percentage change(APC)and average annual percentage change(AAPC)with 95％confidence in-terval(CI)for analyzing trends of thyroid cancer incidence and mortality rates from 2000 to 2021.[Results]The crude incidence rate of thyroid cancer increased from 3.62/105 in 2000 to 79.66/105 in 2021,and ASIRC rose from 3.11/105 to 69.49/105(AAPC=15.55％,95％CI:14.65％～16.61％).The AAPC for ASIRC was slightly higher in female(15.54％,95％CI:14.71％～16.49％)than that in male(15.02％,95％CI:13.53％～16.82％).The increase was significantly more pronounced in rural areas(AAPC=23.34％,95％CI:21.48％～25.33％)compared to urban areas(AAPC=14.12％,95％CI:13.15％～15.43％).Among age groups,the age group of 15～44 years old showed the fastest increase in crude incidence rate(AAPC=20.37％,95％CI:18.46％～22.31％),followed by the age group of 45～64 years old(AAPC=18.41％,95％CI:16.65％～20.19％).Between 2000 and 2021,the crude mortality rate of thyroid cancer rose from 0.34/105 to 0.56/105(AAPC=5.52％,95％CI:3.45％～7.70％),the trend of ASMRC was relatively stable.[Conclusion]From 2000 to 2021,the incidence of thyroid cancer in Zhejiang Province increased markedly,especially among females and younger to middle-aged adults,while the age-standardized mortality rate remained stable.

In modern society, sugary foods have become an integral part of many people′s lives. However, excessive sugar consumption has adverse effects on both overall health and oral health, serving as a contributing factor to the global increasing incidence in oral diseases, cardiovascular diseases, cancers, obesity, and diabetes. In response to the health risks related to high-sugar diets, the World Health Organization (WHO) and World Dental Federation (FDI) have proposed initiatives and recommendations, with various governments implementing different policies and strategies to reduce sugar intake. Chinese government has also taken proactive measures. The "Healthy China Action (2019-2030)" initiative introduced by the State Council in 2019 established a crucial benchmark in limiting the average daily intake of added sugar to 25 g per person forward to 2030. Experts from Chinese Center for Disease Control and Prevention and the field of oral health have meticulously examined the impacts of sugar reduction on oral health, as well as strategies, methods, and practical considerations related to reducing sugar intake through several meeting and wrote the "Expert consensus: reducing free-sugar for caries prevention", which was subsequently reviewed and revised based on the feedback from multiple stakeholders. They have conducted thorough analyses of global trends in sugar reduction and best practices to provide valuable insights to China for crafting effective policies and strategies on sugar reduction. This consensus mainly includes the classification of free sugars, the latest scientific evidence on dental caries, recommendations from WHO on sugar-sweetened beverage taxes, nutrition labeling, advertising, food reform, adjusting supply systems, education, and promotion strategies, as well as sugar reduction actions taken by various governments around the world. Combining the actual situation in China, policy recommendations and authoritative popular science knowledge on sugar reduction for caries prevention to public are proposed to advocate for experts in multiple fields to focus on sugar reduction for caries prevention, promote the work process, and provide the scientific basis for oral health educators.

[Purpose]To analyze the incidence and mortality of thyroid cancer in Zhejiang cancer registration areas in 2021 and trends from 2000 to 2021.[Methods]Based on cancer data from 22 registries across Zhejiang Province between 2000 and 2021,the crude incidence/mortality rates,age-standardized incidence/mortality rates of thyroid cancer by Chinese and world standard population(ASIRC/ASMRC,ASIRW/ASMRW),cumulative rate(0～74 years old)and truncated rate(35～64 years old)were analyzed.Joinpoint regression model was employed to calculate the annual percentage change(APC)and average annual percentage change(AAPC)with 95％confidence in-terval(CI)for analyzing trends of thyroid cancer incidence and mortality rates from 2000 to 2021.[Results]The crude incidence rate of thyroid cancer increased from 3.62/105 in 2000 to 79.66/105 in 2021,and ASIRC rose from 3.11/105 to 69.49/105(AAPC=15.55％,95％CI:14.65％～16.61％).The AAPC for ASIRC was slightly higher in female(15.54％,95％CI:14.71％～16.49％)than that in male(15.02％,95％CI:13.53％～16.82％).The increase was significantly more pronounced in rural areas(AAPC=23.34％,95％CI:21.48％～25.33％)compared to urban areas(AAPC=14.12％,95％CI:13.15％～15.43％).Among age groups,the age group of 15～44 years old showed the fastest increase in crude incidence rate(AAPC=20.37％,95％CI:18.46％～22.31％),followed by the age group of 45～64 years old(AAPC=18.41％,95％CI:16.65％～20.19％).Between 2000 and 2021,the crude mortality rate of thyroid cancer rose from 0.34/105 to 0.56/105(AAPC=5.52％,95％CI:3.45％～7.70％),the trend of ASMRC was relatively stable.[Conclusion]From 2000 to 2021,the incidence of thyroid cancer in Zhejiang Province increased markedly,especially among females and younger to middle-aged adults,while the age-standardized mortality rate remained stable.

Objective:To construct a questionnaire to measure parents′ behavioral and social drivers (BeSD) in administering rotavirus vaccine to their children, and to test the reliability and validity of the questionnaire.Methods:A questionnaire was constructed to investigate the BeSD of rotavirus vaccination among parents based on the WHO BeSD questionnaire. A survey was conducted among parents of children aged 0-3 years old in vaccination clinics from 15 community health service centers in Nanjing City of Jiangsu Province, Yuhuan and Kaihua Counties of Zhejiang Province, Anqing City of Anhui Province, and Yangpu District of Shanghai City from April to May in 2024. Exploratory factor analysis (EFA) and confirmatory factor analysis (CFA) were used to analyze the reliability (internal consistency reliability, combination reliability and split half reliability) and validity (structural validity, convergence validity and discriminative validity) of the questionnaire.Results:A total of 850 questionnaires were collected, of which 614 were valid, with an effective response rate of 72.20%. The EFA resulted in the refinement of the rotavirus vaccine BeSD questionnaire from 37 to 35 items (including 24 core items), and suggested a five-factor structure including the perceptions of vaccine, confidence in vaccine, social process of vaccination, motivation for vaccination, and practical problems of vaccination. The cumulative variance contribution rate reached 56.27%. The CFA confirmed an ideal five-factor model fit (GFI=0.82,CFI=0.86,PNFI=0.67,RMSEA=0.07). The AVE of each dimension was greater than 0.50. The AVE square root of each dimension of the questionnaire was greater than its correlation coefficient. The Cronbach′s α coefficient of the total questionnaire was 0.79.Conclusion:The developed BeSD questionnaire of rotavirus vaccine has good reliability and validity, and can be used to measure parents′ behavioral and social factors in administering rotavirus vaccine to their children.

Objective To study the role of curcumin-mediated sonodynamic therapy in the treatment of malignant melanoma,and to provide a new strategy for the treatment of malignant melanoma.Methods The ultrasonic sound and vibration method was applied to coat curcumin with mouse melanoma cell membrane,thereby forming biomimetic curcumin.The morphology of biomimetic curcumin was observed by transmission electron microscope.Flow cytometry was used to analyze the effect of biomimetic curcumin in terms of in vitro targeting,apoptosis,and intracellular reactive oxygen species(ROS)production.The in vivo experiment was divided into control group,US group,turmeric group,imitation turmeric group,and imitation turmeric+US group,with 3 mice in each group.The in vivo safety of biomimetic curcumin was evaluated by HE staining.In addition,HE,CD31,Ki67,and TUNEL stainings were performed to evaluate the in vivo anti-melanoma therapeutic effect of ultrasound combined with biomimetic curcumin.Results The biomimetic curcumin had a generally uniform morphology and possessed a core-shell structure.Flow cytometry analysis performed with FlowJo showed that the biomimetic curcumin could be effectively taken up by melanoma cells.The apoptosis rate was(10.30±0.61)％in the control group,(10.41±3.13)％in the ultrasound group,(24.97±1.38)％in the curcumin group,(31.39±3.84)％in the biomimetic curcumin group,and(40.89±0.79)％in the biomimetic curcumin and ultrasound combination group.The apoptosis rate in the biomimetic curcumin and ultrasound combination group was higher than those in the other groups(P＜0.05).The results of ROS flow cytometry showed that,compared with the control group,the ultrasound group demonstrated almost no increase in the fluorescence intensity,while the other groups showed an increase in the fluorescence intensity to varying degrees.There was no significant difference in the fluorescence intensity between the biomimetic curcumin group([1.10±0.38]％)and the curcumin group([0.73±0.26]％)(P＞0.05).The fluorescence intensity of the biomimetic curcumin and ultrasound combination group([3.35±0.04]％)was higher than those of the other groups(P＜0.05).HE staining showed no obvious abnormalities in the morphology of heart,liver,spleen,lung,and kidney tissues in any of the treatment groups.HE staining showed the most significant changes in cell morphology in the biomimetic curcumin and ultrasound combination group,followed by the biomimetic curcumin group and the curcumin group.No obvious abnormalities in tumor cell morphology were observed in the ultrasound group.According to the respective results of CD31 staining,Ki67 staining,and TUNEL staining,the biomimetic curcumin and ultrasound combination group had the largest brown area,the highest number of red fluorescence,and the highest number of green fluorescence,followed by the biomimetic curcumin group and the curcumin group.Conclusion The biomimetic curcumin displays uniform morphology,a core-shell structure,and good targeting properties.When it is used in combination with ultrasound,biomimetic curcumin demonstrates a good anti-tumor therapeutic effect both in vivo and in vitro.

Important forensic diagnostic indicators of sudden death in coronary atherosclerotic heart dis-ease,such as acute or chronic myocardial ischemic changes,sometimes make it difficult to locate the ischemic site due to the short death process,the lack of tissue reaction time.In some cases,the de-ceased died of sudden death on the first-episode,resulting in difficulty for medical examiners to make an accurate diagnosis.However,clinical studies on coronary instability plaque revealed the key role of coronary spasm and thrombosis caused by their lesions in sudden coronary death process.This paper mainly summarizes the pathological characteristics of unstable coronary plaque based on clinical medi-cal research,including plaque rupture,plaque erosion and calcified nodules,as well as the influencing factors leading to plaque instability,and briefly describes the research progress and technique of the atherosclerotic plaques,in order to improve the study on the mechanism of sudden coronary death and improve the accuracy of the forensic diagnosis of sudden coronary death by diagnosing different patho-logic states of coronary atherosclerotic plaques.

OBJECTIVES: To estimate postmortem interval (PMI) by analyzing the protein changes in skeletal muscle tissues with the protein chip technology combined with multivariate analysis methods. METHODS: Rats were sacrificed for cervical dislocation and placed at 16 ℃. Water-soluble proteins in skeletal muscles were extracted at 10 time points (0 d, 1 d, 2 d, 3 d, 4 d, 5 d, 6 d, 7 d, 8 d and 9 d) after death. Protein expression profile data with relative molecular mass of 14 000-230 000 were obtained. Principal component analysis (PCA) and orthogonal partial least squares (OPLS) were used for data analysis. Fisher discriminant model and back propagation (BP) neural network model were constructed to classify and preliminarily estimate the PMI. In addition, the protein expression profiles data of human skeletal muscles at different time points after death were collected, and the relationship between them and PMI was analyzed by heat map and cluster analysis. RESULTS: The protein peak of rat skeletal muscle changed with PMI. The result of PCA combined with OPLS discriminant analysis showed statistical significance in groups with different time points (P<0.05) except 6 d, 7 d and 8 d after death. By Fisher discriminant analysis, the accuracy of internal cross-validation was 71.4% and the accuracy of external validation was 66.7%. The BP neural network model classification and preliminary estimation results showed the accuracy of internal cross-validation was 98.2%, and the accuracy of external validation was 95.8%. There was a significant difference in protein expression between 4 d and 25 h after death by the cluster analysis of the human skeletal muscle samples. CONCLUSIONS The protein chip technology can quickly, accurately and repeatedly obtain water-soluble protein expression profiles in rats' and human skeletal muscles with the relative molecular mass of 14 000-230 000 at different time points postmortem. The establishment of multiple PMI estimation models based on multivariate analysis can provide a new idea and method for PMI estimation.
Animals ; Humans ; Rats ; Multivariate Analysis ; Postmortem Changes ; Protein Array Analysis ; Technology