1.Current status and influencing factors of knowledge-attitude-practice in myopia prevention and control among children and adolescents in Ningbo
Jue WANG ; Xiaotian LIU ; Xia JIN ; Yanli ZHANG ; Hongjun LI ; Honger SUN ; Aiai CHEN ; Yuan TANG
International Eye Science 2026;26(3):518-522
AIM:To investigate the current status and influencing factors of knowledge-attitude-practice in myopia prevention and control among children and adolescents in Ningbo City, thereby providing a scientific basis for formulating targeted prevention strategies.METHODS: Children and adolescents aged 6-12 years old were selected from the medical-school collaborative myopia prevention network in Ningbo City between August 2024 and May 2025 using stratified cluster sampling. Information on myopia prevention knowledge(15 items)and practice(9 items)was collected through questionnaire surveys. Logistic regression models were used to analyze factors influencing myopia occurrence in children and adolescents.RESULTS: A total of 664 children and adolescents aged 6-12 years were enrolled in this study. Participants were divided by age into three groups: 6-7 years old(n=221), 8-9 years old(n=221), and 10-12 years old(n=222). Of the 664 questionnaires distributed, 637 valid questionnaires were returned(201 from the 6-7 age group, 235 from the 8-9 age group, and 201 from the 10-12 age group), yielding an effective response rate of 95.9%. Based on myopia screening results, the non-myopic group comprised 203 participants(31.9%), including 100 males and 103 females, with a mean age of 8.82±1.98 years old. The myopic group comprised 434 participants(68.1%), including 213 males and 221 females, with a mean age of 9.10±1.95 years old. The myopia prevalence rates in the 6-7, 8-9, and 10-12 age groups were 37.8%(76/201), 71.9%(169/235), and 94.0%(189/201), respectively(P<0.001). Regarding the knowledge and practice of myopia prevention, the overall awareness rate in the non-myopic group(59.7%±9.7%)was significantly higher than that in the myopic group(48.7%±8.5%; P<0.001). Additionally, the non-myopic group scored higher on the key practice of “regular eye examinations”(4.27±0.96)compared to the myopic group(4.10±1.05; P<0.05). Logistic regression analysis indicated that age was the primary risk factor for myopia occurrence.CONCLUSION: Age is the dominant factor in the onset of myopia, and there is a phenomenon of “knowledge-practice gap”; the traditional health education model has limitations, and a precise prevention and control system based on developmental patterns should be established.
2.Expert Consensus on Clinical Application of Qinbaohong Zhike Oral Liquid in Treatment of Acute Bronchitis and Acute Attack of Chronic Bronchitis
Jian LIU ; Hongchun ZHANG ; Chengxiang WANG ; Hongsheng CUI ; Xia CUI ; Shunan ZHANG ; Daowen YANG ; Cuiling FENG ; Yubo GUO ; Zengtao SUN ; Huiyong ZHANG ; Guangxi LI ; Qing MIAO ; Sumei WANG ; Liqing SHI ; Hongjun YANG ; Ting LIU ; Fangbo ZHANG ; Sheng CHEN ; Wei CHEN ; Hai WANG ; Lin LIN ; Nini QU ; Lei WU ; Dengshan WU ; Yafeng LIU ; Wenyan ZHANG ; Yueying ZHANG ; Yongfen FAN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(4):182-188
The Expert Consensus on Clinical Application of Qinbaohong Zhike Oral Liquid in Treatment of Acute Bronchitis and Acute Attack of Chronic Bronchitis (GS/CACM 337-2023) was released by the China Association of Chinese Medicine on December 13th, 2023. This expert consensus was developed by experts in methodology, pharmacy, and Chinese medicine in strict accordance with the development requirements of the China Association of Chinese Medicine (CACM) and based on the latest medical evidence and the clinical medication experience of well-known experts in the fields of respiratory medicine (pulmonary diseases) and pediatrics. This expert consensus defines the application of Qinbaohong Zhike oral liquid in the treatment of cough and excessive sputum caused by phlegm-heat obstructing lung, acute bronchitis, and acute attack of chronic bronchitis from the aspects of applicable populations, efficacy evaluation, usage, dosage, drug combination, and safety. It is expected to guide the rational drug use in medical and health institutions, give full play to the unique value of Qinbaohong Zhike oral liquid, and vigorously promote the inheritance and innovation of Chinese patent medicines.
3.Systematic review on the extracellular vesicles in reproductive medicine and gamete union.
Yutao WANG ; Honghao SUN ; Fangdie YE ; Zhiwei LI ; Zhongru FAN ; Xun FU ; Yi LU ; Jianbin BI ; Hongjun LI
Journal of Pharmaceutical Analysis 2025;15(10):101261-101261
In this comprehensive review, we delve into the evolution of drug delivery systems in reproductive medicine with a focus on the emerging role of exosomes, a class of extracellular vesicles. Exosomes offer unique advantages in overcoming these challenges due to their inherent biocompatibility, stability, and ability to facilitate targeted delivery. This review provides a detailed examination of exosome biogenesis and their function in cellular communication, setting the stage for understanding their potential as drug delivery vehicles. We explore the mechanisms through which exosomes can be loaded with small molecule drugs and the benefits they offer over synthetic nanoparticles. The review highlights groundbreaking case studies that illustrate the successful application of exosome-mediated drug delivery in reproductive health, including enhancing fertility treatments, supporting gamete and embryo development, and facilitating maternal-fetal communication. This study aims to provide a precise understanding of how exosomal drug delivery can revolutionize treatments for reproductive health disorders, paving the way for future therapeutic applications. Lastly, we touch upon the promising therapeutic implications of exosomal delivery for proteins and genes, offering a window into future treatments for reproductive health disorders.
4.Dimeric sesquiterpenoids with anti-inflammatory activities from Inula britannica.
Juan ZHANG ; Jiankun YAN ; Hongjun DONG ; Rui ZHANG ; Jing CHANG ; Yanli FENG ; Xinrong XU ; Wei LI ; Feng QIU ; Chengpeng SUN
Chinese Journal of Natural Medicines (English Ed.) 2025;23(8):961-971
In continuation of research aimed at identifying anti-inflammatory agents from natural sesquiterpenoids, an activity-guided fractionation approach utilizing lipopolysaccharide (LPS)-mediated RAW264.7 cells was employed to investigate chemical constituents from Inula Britannica (I. britannica). Seven novel sesquiterpenoid dimers inulabritanoids A-G (1-7) and two novel sesquiterpenoid monomers inulabritanoids H (8) and I (9) were isolated from I. britannica together with eighteen known compounds (10-27). The structural elucidation was accomplished through comprehensive analysis of 1D and 2D nuclear magnetic resonance (NMR), high-resolution mass spectrometry (HR-MS), and electronic circular dichroism (ECD) spectra, complemented by quantum chemical calculations. Compounds 1, 2, 12, 16, 19, and 26 demonstrated inhibitory effects on NO production, with IC50 values of 3.65, 5.48, 3.29, 6.91, 3.12, and 5.67 μmol·L-1, respectively. Mechanistic studies revealed that compound 1 inhibited IκB kinase β (IKKβ) phosphorylation, thereby blocking nuclear factor κB (NF-κB) nuclear translocation, and activated the kelch-like ECH-associated protein 1 (Keap1)/nuclear factor erythroid 2-related factor 2 (Nrf2) signal pathway, leading to decreased expression of NADPH oxidase 2 (NOX-2), inducible nitric oxide synthase (iNOS), tumor necrosis factor α (TNF-α), interleukin-6 (IL-6), monocyte chemotactic protein-1 (MCP-1), IL-1β, and IL-1α and increased expression of NAD(P)H: quinone oxidoreductase 1 (NQO-1) and heme oxygenase-1 (HO-1), thus exhibiting anti-inflammatory effects in vitro. These results indicate that dimeric sesquiterpenoids may serve as promising candidates for anti-inflammatory drug development.
Mice
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Animals
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Sesquiterpenes/isolation & purification*
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Anti-Inflammatory Agents/isolation & purification*
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Inula/chemistry*
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RAW 264.7 Cells
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Nitric Oxide
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Molecular Structure
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NF-kappa B/immunology*
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NF-E2-Related Factor 2/immunology*
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Macrophages/immunology*
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Nitric Oxide Synthase Type II/immunology*
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Plant Extracts/pharmacology*
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Lipopolysaccharides
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Tumor Necrosis Factor-alpha/immunology*
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I-kappa B Kinase/genetics*
5.Levels of peripheral blood lung cancer biomarkers in apparently healthy individuals in Beijing and surrounding areas and establishment and validation of reference intervals
Xinmiao LIU ; Ping SUN ; Mingyang HE ; Yan KANG ; Xiaoying LOU ; Yue WU ; Yulin SUN ; Hongjun GAO
Cancer Research and Clinic 2025;37(6):412-421
Objective:To explore the features of levels of lung cancer biomarkers in peripheral blood of adults in Beijing and surrounding areas, and establish personalized reference intervals for these biomarkers.Methods:A cross sectional study was conducted. The lung cancer biomarker data, including carcinoembryonic antigen (CEA), cytokeratin 19 fragment antigen 21-1 (CYFRA21-1), neuron specific enolase (NSE), progastrin-releasing peptide (ProGRP), and squamous cell carcinoma antigen (SCC-Ag), collected from adults who underwent cancer prevention examinations at the Cancer Hospital of the Chinese Academy of Medical Sciences from July 2021 to July 2022 were retrospectively analyzed. The interquartile range method was used to eliminate outliers, and the P95 value was calculated. Upper limit of 5 lung cancer biomarkers in different gender and age groups were obtained by referring to the reference intervals of quantitative analytes in the clinical laboratory (WS/T 402-2024). By analyzing the data of 208 adults who underwent cancer prevention physical examinations at the same center in June 2021 and 140 patients with benign lung masses confirmed by surgical resection pathology from January 2016 to June 2022, the established reference intervals for biomarkers were validated. Results:Two thousand six hundred and twenty-six cases of apparently healthy physical examiners were included for constructing reference intervals, including 1 456 males (55.4%) and 1 170 females (44.6%); the age range was 20-88 years old. The serum levels [ M ( Q1, Q3)] of CEA, NSE, ProGRP, SCC-Ag and CYFRA21-1 in 2 626 cases were 1.63 (1.07, 2.43) ng/ml, 13.08 (11.44, 14.77) ng/ml, 34.93 (29.02, 42.19) pg/ml, 0.80 (0.60, 1.00) ng/ml and 1.96 (1.48, 2.63) ng/ml, respectively. The serum levels of CEA [1.88 (1.22, 2.76) ng/ml vs. 1.41 (0.93, 2.02) ng/ml], NSE [13.31 (11.87, 15.00) ng/ml vs. 12.69 (10.96, 14.53) ng/ml], SCC-Ag [0.9 (0.7, 1.1) ng/ml vs. 0.7 (0.6, 0.9) ng/ml], and CYFRA21-1 [2.02 (1.53, 2.71) ng/ml vs. 1.87 (1.40, 2.51) ng/ml] in males were higher than those in females, and ProGRP [34.00 (28.25, 41.55) pg/ml vs. 36.12 (29.97, 42.98) pg/ml] was lower than that in females, and the differences were statistically significant (all P < 0.001). There were statistically significant differences in serum CEA levels between the groups of ≤ 40 years old (458 cases), >40-50 years old (827 cases), >50-60 years old (783 cases), >60-70 years old (412 cases), and >70 years old (146 cases) in pairwise comparison (all P < 0.05). Except for the age groups of ≤ 40 years old and >40-50 years old and the age groups of >60-70 years old and >70 years old, there were statistically significant differences in serum NSE levels among the other age groups in pairwise comparison (all P < 0.05). There were statistically significant differences in serum ProGRP levels between the 5 age groups (all P < 0.05). There were statistically significant differences when comparing the serum SCC-Ag level in the >40-50 age group, >50-60 age group and >60-70 age group with that in the ≤40 age group and >70 age group, respectively (all P < 0.05). However, there was no statistically significant difference between the other age groups in pairwise comparison (all P > 0.05). There were statistically significant differences in serum CYFRA21-1 levels between the 5 age groups (all P < 0.05). When gender and age were not distinguished, the P95 values of serum CEA, NSE, ProGRP, SCC-Ag and CYFRA21-1 levels were 4.44 ng/ml, 16.61 ng/ml, 57.65 pg/ml, 1.50 ng/ml, and 4.21 ng/ml, respectively. Considering gender and age, except for the >70 age group with no statistically significant difference in the P95 value of serum CEA level between males and females ( P > 0.05), the P95 value of serum CEA level in males was higher than that in females in all other age groups (all P < 0.001); the P95 values of serum CEA level in both males and females increased with age, but showed a decreasing trend in males over the age of 70. The P95 value of serum NSE level in males was higher than that in females in the age groups of ≤ 40 years and >40-50 years (both P < 0.05), while there was no statistically significant difference in the P95 value of serum NSE level between males and females in other age groups (all P > 0.05). The P95 values of serum NSE level in both males and females decreased firstly and increased later with age, reaching their highest levels at the age of >70. The P95 values of serum ProGRP level in females aged ≤ 40 and >50-60 were higher than those in males (both P < 0.05), while there was no statistically significant difference in the P95 value of serum ProGRP level between genders in other age groups (all P > 0.05); the P95 values of serum ProGRP level in both males and females increased with age. There was no statistically significant difference in the P95 value of serum SCC-Ag level between males and females in the ≤ 40 age group ( P > 0.05), while the P95 value of serum SCC-Ag level in males was higher than that in females in all other age groups (all P < 0.05). The P95 values of serum SCC-Ag level in males increased with age, while they were stable in females. There was no statistically significant difference in the P95 value of serum CYFRA21-1 level between males and females in the >60-70 age group ( P > 0.05), while the P95 value of serum CYFRA21-1 level in males was higher than those in females in all other age groups (all P < 0.05); the P95 values of serum CYFRA21-1 level in both males and females increased with age. Based on data from 2 626 apparently healthy physical examiners, reference intervals for the levels of 5 lung cancer biomarkers were constructed in different age groups of different genders. Validation was conducted on 208 physical examiners and 140 patients with benign lung lesions, and it was found that the compliance rate of using newly created reference intervals for different gender and age groups to interpret detection results was >90%, and the validation was passed. Conclusions:There are gender and age differences in the reference intervals of CEA, CYFRA21-1, NSE, ProGRP, and SCC-Ag in peripheral blood of adults in Beijing and surrounding areas. The constructed reference intervals of gender and age for biomarkers have been validated and shown good results, providing reference for optimizing the clinical application of lung cancer-related biomarkers.
6.Preparation and immunogenicity evaluation of rotavirus VP8-mRNA vaccine
Qingmei LENG ; Xianqiong TANG ; Rong CHEN ; Xiaoqing HU ; Xiaopeng SONG ; Yan LI ; Jinmei LI ; Lida YAO ; Xiaochen LIN ; Jinyuan WU ; Maosheng SUN ; Hongjun LI ; Yan ZHOU
Chinese Journal of Microbiology and Immunology 2025;45(9):727-732
Objective:To construct a VP8-mRNA vaccine using human rotavirus spike protein VP8 domain as the immunogen and analyze its immunogenicity in mice.Methods:The VP8-mRNA sequence was designed, optimized, and synthesized. The VP8 gene of rotavirus G1P[8] type was used to construct the plasmid pUC57-VP8-Kan-SapⅠ, which was then sequenced. The plasmid confirmed by sequencing was subjected to large-scale amplification and extraction, followed by linearization, in vitro transcription, and capping. The purified capped products were encapsulated with lipid nanoparticles using a microfluidic control apparatus. The encapsulated VP8-mRNA vaccine was administered intramuscularly to mice at 10, 15, and 20 μg. Serum samples were collected for antibody detection by ELISA. Cellular immune responses were detected by flow cytometry and ELISPOT. Statistical analysis was performed using one-way or two-way analysis of variance and Tukey-Kramer test. Results:The encapsulated VP8-mRNA vaccine was rounded and spherical, with a particle size of about 100 nm, a polymer dispersion index of 0.088, and an encapsulation rate of 92.3%. Two doses of VP8-mRNA vaccine immunization could induce a good immune response in mice. The level of IgG antibody induced after immunization in the 15 μg group was comparable to that of the 20 μg group, and there was no statistical difference ( P>0.05), but the antibody levels in the two groups were significantly higher than that in the 10 μg group ( P<0.000 1). VP8-mRNA vaccine could induce neutralizing antibodies against rotavirus G1 and G9 types. The highest level of neutralizing antibodies against rotavirus type G1 was observed in the 15 μg group, which was significantly higher than that in the 10 μg group ( P<0.05). All immunization groups exhibited good neutralizing ability against rotavirus G9 type. The results of ELISPOT showed that lymphocytes from mice in each vaccine group were able to secrete IFN-γ when stimulated with VP8 peptide. Flow cytometry showed that the proportions of CD8 + T cell subsets in the vaccine groups were higher than that in the control group. Conclusion:The VP8-mRNA vaccine has good immunogenicity in mice and can induce good humoral and T-cell immune responses.
7.Expression and significance of 25-hydroxyvitamin D and immune cytokine in patients with unexplained recurrent spontaneous abortion
Xiaoli MU ; Zhaoliang SUN ; Guangsong LIU ; Peng YU ; Hongjun YANG
Journal of Clinical Medicine in Practice 2025;29(15):79-82
Objective To investigate the expression and significance of 25-hydroxyvitamin D[25(OH)D],immune cell subsets,and helper T cell(Th)1 and Th2 in patients with unexplained recurrent spontaneous abortion(URSA).Methods A total of 307 URSA patients were enrolled as URSA group,which was further divided into non-maintenance of pregnancy group(n=127)and ma-intenance of pregnancy group(n=180)based on pregnancy maintenance status.Another 307 healthy pregnant women in the same period were selected as control group.The levels of 25(OH)D,immune cell subsets,and Th1/Th2 expression were compared among these groups.Pearson correlation analy-sis was used to explore the correlations of 25(OH)D and immune cell subsets with Th1/Th2 expres-sion in URSA patients.Results Compared with the control group,the URSA group had significantly lower levels of 25(OH)D and interleukin-10(IL-10),and higher levels of CD3+,CD3+CD4+,CD3+CD8+,CD4+/CD8+,CD16+CD56+,CD19+,interleukin-2(IL-2),and tumor necrosis fac-tor-α(TNF-α)(P<0.001).Compared with the maintenance of pregnancy group,the non-maintenance of pregnancy group had significantly lower levels of 25(OH)D and IL-10,and higher levels of CD3+,CD3+CD4+,CD3+CD8+,CD4+/CD8+,CD16+CD56+,CD19+,IL-2,and TNF-α(P<0.001).Pearson analysis revealed that 25(OH)D in URSA patients was significantly negatively correlated with IL-2 and TNF-α(P<0.05),while positively correlated with IL-10(P<0.05).Immune cell sub-sets,including CD3+,CD3+CD4+,CD3+CD8+,CD4+/CD8+,CD16+CD56+,and CD19+were significantly positively correlated with IL-2 and TNF-α(P<0.05),while negatively correlated with IL-10 in URSA patients(P<0.05).Conclusion There are abnormalities in 25(OH)D,im-mune cell subsets,and Th1/Th2 cytokine expression in URSA patients,and these expressions differ in URSA patients with different pregnancy outcomes.The underlying mechanism may be related to the regulation of Th1/Th2 cytokine balance.
8.Preparation and immunogenicity evaluation of rotavirus VP8-mRNA vaccine
Qingmei LENG ; Xianqiong TANG ; Rong CHEN ; Xiaoqing HU ; Xiaopeng SONG ; Yan LI ; Jinmei LI ; Lida YAO ; Xiaochen LIN ; Jinyuan WU ; Maosheng SUN ; Hongjun LI ; Yan ZHOU
Chinese Journal of Microbiology and Immunology 2025;45(9):727-732
Objective:To construct a VP8-mRNA vaccine using human rotavirus spike protein VP8 domain as the immunogen and analyze its immunogenicity in mice.Methods:The VP8-mRNA sequence was designed, optimized, and synthesized. The VP8 gene of rotavirus G1P[8] type was used to construct the plasmid pUC57-VP8-Kan-SapⅠ, which was then sequenced. The plasmid confirmed by sequencing was subjected to large-scale amplification and extraction, followed by linearization, in vitro transcription, and capping. The purified capped products were encapsulated with lipid nanoparticles using a microfluidic control apparatus. The encapsulated VP8-mRNA vaccine was administered intramuscularly to mice at 10, 15, and 20 μg. Serum samples were collected for antibody detection by ELISA. Cellular immune responses were detected by flow cytometry and ELISPOT. Statistical analysis was performed using one-way or two-way analysis of variance and Tukey-Kramer test. Results:The encapsulated VP8-mRNA vaccine was rounded and spherical, with a particle size of about 100 nm, a polymer dispersion index of 0.088, and an encapsulation rate of 92.3%. Two doses of VP8-mRNA vaccine immunization could induce a good immune response in mice. The level of IgG antibody induced after immunization in the 15 μg group was comparable to that of the 20 μg group, and there was no statistical difference ( P>0.05), but the antibody levels in the two groups were significantly higher than that in the 10 μg group ( P<0.000 1). VP8-mRNA vaccine could induce neutralizing antibodies against rotavirus G1 and G9 types. The highest level of neutralizing antibodies against rotavirus type G1 was observed in the 15 μg group, which was significantly higher than that in the 10 μg group ( P<0.05). All immunization groups exhibited good neutralizing ability against rotavirus G9 type. The results of ELISPOT showed that lymphocytes from mice in each vaccine group were able to secrete IFN-γ when stimulated with VP8 peptide. Flow cytometry showed that the proportions of CD8 + T cell subsets in the vaccine groups were higher than that in the control group. Conclusion:The VP8-mRNA vaccine has good immunogenicity in mice and can induce good humoral and T-cell immune responses.
9.Levels of peripheral blood lung cancer biomarkers in apparently healthy individuals in Beijing and surrounding areas and establishment and validation of reference intervals
Xinmiao LIU ; Ping SUN ; Mingyang HE ; Yan KANG ; Xiaoying LOU ; Yue WU ; Yulin SUN ; Hongjun GAO
Cancer Research and Clinic 2025;37(6):412-421
Objective:To explore the features of levels of lung cancer biomarkers in peripheral blood of adults in Beijing and surrounding areas, and establish personalized reference intervals for these biomarkers.Methods:A cross sectional study was conducted. The lung cancer biomarker data, including carcinoembryonic antigen (CEA), cytokeratin 19 fragment antigen 21-1 (CYFRA21-1), neuron specific enolase (NSE), progastrin-releasing peptide (ProGRP), and squamous cell carcinoma antigen (SCC-Ag), collected from adults who underwent cancer prevention examinations at the Cancer Hospital of the Chinese Academy of Medical Sciences from July 2021 to July 2022 were retrospectively analyzed. The interquartile range method was used to eliminate outliers, and the P95 value was calculated. Upper limit of 5 lung cancer biomarkers in different gender and age groups were obtained by referring to the reference intervals of quantitative analytes in the clinical laboratory (WS/T 402-2024). By analyzing the data of 208 adults who underwent cancer prevention physical examinations at the same center in June 2021 and 140 patients with benign lung masses confirmed by surgical resection pathology from January 2016 to June 2022, the established reference intervals for biomarkers were validated. Results:Two thousand six hundred and twenty-six cases of apparently healthy physical examiners were included for constructing reference intervals, including 1 456 males (55.4%) and 1 170 females (44.6%); the age range was 20-88 years old. The serum levels [ M ( Q1, Q3)] of CEA, NSE, ProGRP, SCC-Ag and CYFRA21-1 in 2 626 cases were 1.63 (1.07, 2.43) ng/ml, 13.08 (11.44, 14.77) ng/ml, 34.93 (29.02, 42.19) pg/ml, 0.80 (0.60, 1.00) ng/ml and 1.96 (1.48, 2.63) ng/ml, respectively. The serum levels of CEA [1.88 (1.22, 2.76) ng/ml vs. 1.41 (0.93, 2.02) ng/ml], NSE [13.31 (11.87, 15.00) ng/ml vs. 12.69 (10.96, 14.53) ng/ml], SCC-Ag [0.9 (0.7, 1.1) ng/ml vs. 0.7 (0.6, 0.9) ng/ml], and CYFRA21-1 [2.02 (1.53, 2.71) ng/ml vs. 1.87 (1.40, 2.51) ng/ml] in males were higher than those in females, and ProGRP [34.00 (28.25, 41.55) pg/ml vs. 36.12 (29.97, 42.98) pg/ml] was lower than that in females, and the differences were statistically significant (all P < 0.001). There were statistically significant differences in serum CEA levels between the groups of ≤ 40 years old (458 cases), >40-50 years old (827 cases), >50-60 years old (783 cases), >60-70 years old (412 cases), and >70 years old (146 cases) in pairwise comparison (all P < 0.05). Except for the age groups of ≤ 40 years old and >40-50 years old and the age groups of >60-70 years old and >70 years old, there were statistically significant differences in serum NSE levels among the other age groups in pairwise comparison (all P < 0.05). There were statistically significant differences in serum ProGRP levels between the 5 age groups (all P < 0.05). There were statistically significant differences when comparing the serum SCC-Ag level in the >40-50 age group, >50-60 age group and >60-70 age group with that in the ≤40 age group and >70 age group, respectively (all P < 0.05). However, there was no statistically significant difference between the other age groups in pairwise comparison (all P > 0.05). There were statistically significant differences in serum CYFRA21-1 levels between the 5 age groups (all P < 0.05). When gender and age were not distinguished, the P95 values of serum CEA, NSE, ProGRP, SCC-Ag and CYFRA21-1 levels were 4.44 ng/ml, 16.61 ng/ml, 57.65 pg/ml, 1.50 ng/ml, and 4.21 ng/ml, respectively. Considering gender and age, except for the >70 age group with no statistically significant difference in the P95 value of serum CEA level between males and females ( P > 0.05), the P95 value of serum CEA level in males was higher than that in females in all other age groups (all P < 0.001); the P95 values of serum CEA level in both males and females increased with age, but showed a decreasing trend in males over the age of 70. The P95 value of serum NSE level in males was higher than that in females in the age groups of ≤ 40 years and >40-50 years (both P < 0.05), while there was no statistically significant difference in the P95 value of serum NSE level between males and females in other age groups (all P > 0.05). The P95 values of serum NSE level in both males and females decreased firstly and increased later with age, reaching their highest levels at the age of >70. The P95 values of serum ProGRP level in females aged ≤ 40 and >50-60 were higher than those in males (both P < 0.05), while there was no statistically significant difference in the P95 value of serum ProGRP level between genders in other age groups (all P > 0.05); the P95 values of serum ProGRP level in both males and females increased with age. There was no statistically significant difference in the P95 value of serum SCC-Ag level between males and females in the ≤ 40 age group ( P > 0.05), while the P95 value of serum SCC-Ag level in males was higher than that in females in all other age groups (all P < 0.05). The P95 values of serum SCC-Ag level in males increased with age, while they were stable in females. There was no statistically significant difference in the P95 value of serum CYFRA21-1 level between males and females in the >60-70 age group ( P > 0.05), while the P95 value of serum CYFRA21-1 level in males was higher than those in females in all other age groups (all P < 0.05); the P95 values of serum CYFRA21-1 level in both males and females increased with age. Based on data from 2 626 apparently healthy physical examiners, reference intervals for the levels of 5 lung cancer biomarkers were constructed in different age groups of different genders. Validation was conducted on 208 physical examiners and 140 patients with benign lung lesions, and it was found that the compliance rate of using newly created reference intervals for different gender and age groups to interpret detection results was >90%, and the validation was passed. Conclusions:There are gender and age differences in the reference intervals of CEA, CYFRA21-1, NSE, ProGRP, and SCC-Ag in peripheral blood of adults in Beijing and surrounding areas. The constructed reference intervals of gender and age for biomarkers have been validated and shown good results, providing reference for optimizing the clinical application of lung cancer-related biomarkers.
10.The role and regulatory mechanism of RNA binding protein ZFP36in hypoxia/reoxygenation injury of cardiomyocytes
Guo LÜ ; Chaofeng SUN ; Hao ZHANG ; Hongjun LI ; Fang WANG
Journal of Xi'an Jiaotong University(Medical Sciences) 2024;45(2):219-227
【Objective】 To explore the role of ZFP36 in cardiomyocyte injury and autophagy induced by hypoxia/reoxygenation (H/R) so as to clarify its molecular regulatory mechanism. 【Methods】 H9C2 rat cardiomyocytes were infected with ZFP36 overexpressing lentivirus (OE-ZFP36) or its negative control lentivirus (OE-ZFP36 NC) to construct stable cell lines, respectively. Transfection of ATG4D overexpression plasmid (OE-ATG4D) improved the expression of ATG4D. Hypoxia/reoxygenation (H/R) induced myocardial cell injury. H9C2 cells were mainly divided into control group, H/R group, OE-ZFP36 NC+H/R group, OE-ZFP36+H/R group, OE-ATG4D NC+H/R group, OE-ATG4D+H/R group, OE-ZFP36+OE-ATG4D NC+H/R group, and OE-ZFP36+OE-ATG4D+H/R group. The protein expressions of ATG4D, Beclin1, LC3 and ZFP36 in H9C2 cells were detected by Western blotting. The mRNA levels of ZFP36 and ATG4D in H9C2 cells were detected by Real-time fluorescence quantitative PCR (qPCR). The viability of H9C2 cells was detected by CCK-8 assay. The levels of interleukin (IL-6) and tumor necrosis factor (TNF-α) in H9C2 cells were detected by enzyme-linked immunosorbent assay (ELISA). Reactive oxygen species (ROS) in H9C2 cells were detected by DCFH-DA method. SOD detection kit was used to detect the SOD level in H9C2 cells. The apoptosis of H9C2 cells was detected by flow cytometry. LC3 autophagosomes in H9C2 cells were detected by cellular immunofluorescence. Dual-luciferase reporter gene assay was used to detect the binding of ZFP36 and ATG4D mRNA in H9C2 cells. 【Results】 Compared with control group, H/R group showed decreased cell viability, increased IL-6 and TNF-α levels, increased ROS levels and decreased SOD levels, increased cell apoptosis. Up-regulated ATG4D and Beclin1 protein expression, increased LC3Ⅱ/LC3Ⅰ ratio, as well as upregulated ZFP36 expression were found in H/R group (all P<0.05). Compared with OE-ZFP36 NC+H/R group, elevated cell viability, decreased IL-6 and TNF-α levels, decreased ROS levels and increased SOD levels, reduced cell apoptosis (P<0.05), and downregulated ATG4D and Beclin1 protein expression, decreased LC3Ⅱ/LC3Ⅰ ratio were shown in OE-ZFP36+H/R group (all P<0.05). Compared with infection with OE-ZFP36 NC lentivirus, infection with OE-ZFP36 lentivirus decreased the luciferase activity of ATG4D 3′-UTR reporter gene, decreased the stability of ATG4D mRNA, and downregulated the H/R-induced ATG4D mRNA expression (all P<0.05). Compared with OE-ATG4D NC+H/R group, OE-ATG4D+H/R group had upregulated ATG4D mRNA and protein expression, decreased cell viability, increased IL-6 and TNF-α levels, increased ROS levels, decreased SOD levels and elevated cell apoptosis (all P<0.05). Compared with OE-ZFP36+OE-ATG4D NC+H/R group, OE-ZFP36+OE-ATG4D+H/R group had decreased cell viability, increased IL-6 and TNF-α levels, increased ROS levels, decreased SOD levels and elevated cell apoptosis (all P<0.05). 【Conclusion】 The expression of ZFP36 is upregulated in H/R-induced cardiomyocyte injury. The overexpression of ZFP36 inhibits H/R-induced cardiomyocyte injury and autophagy by regulating ATG4D, thus resisting cardiomyocyte H/R injury. It proves that ZFP36 is an important regulatory molecule against MI/RI.

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