1.Effect of targeted silencing of DNMT3A on collagen deposition, proliferation and migration activity of mouse lung fibroblasts
Xianchen Wang ; Junbo You ; Hui Ling ; Jiahao Fan ; Qi Chen ; Hui Tao ; Jiming Sha
Acta Universitatis Medicinalis Anhui 2025;60(1):66-72
		                        		
		                        			Objective:
		                        			To investigate the effect of targeted silencing of DNA methyltransferase 3A(DNMT3A) on collagen deposition, proliferation and migration activity of mouse lung fibroblasts(PFs). 
		                        		
		                        			Methods:
		                        			In order to ensure the proliferation and migration activity of primary fibroblasts, the lung tissues of neonatal C57 suckling mice were taken, PFs were extracted after being sheared, and the morphology was observed and identified under the microscope. PFs cells were activated by 5 ng/ml TGF-β1for 24 h after cell attachment, and DNMT3A silencing model was constructed by small interfering RNA; The experiment was divided into control group, TGF-β1group, TGF-β1+ siRNA-NC group and TGF-β1+ siRNA-DNMT3A group. The protein expressions of DNMT3A, α-smooth muscle actin(α-SMA) and Collagen Ⅰ were detected by Western blot; Real time quantitative reverse transcription polymerase chain reaction(RT-qPCR) was used to detect the mRNA expression changes ofDNMT3A,α-SMAandCollagenⅠ. The proliferation ability of PFs was detected by CCK-8 and EdU staining; the migration ability of PFs was detected by scratch test and Transwell migration test. 
		                        		
		                        			Results:
		                        			Compared with the control group, TGF-β1induced the increase of DNMT3A in the activated PFs cell group(P<0.01), the protein and mRNA levels of fibrosis and proliferation related indicators α-SMA and Collagen Ⅰ also increased(allP<0.05), and the proliferation and migration ability of PFs increased(allP<0.000 1). Compared with the siRNA-NC group, the protein expression levels of DNMT3A(P<0.000 1) and related indicators α-SMA(P<0.01) and Collagen Ⅰ(P<0.01) significantly decreased in the DNMT3A silencing group by Western blot, and the mRNA levels ofDNMT3A,α-SMAandCollagenⅠby RT-qPCR also decreased(allP<0.001), and the proliferation(P<0.01) and migration ability(P<0.05) of PFs cells decreased compared with the control group.
		                        		
		                        			Conclusion
		                        			Silencing DNMT3A can inhibit the deposition of collagen and the proliferation of PFs. DNMT3A can promote the proliferation and migration of PFs, and then promote the activation of PFs and the development of pulmonary fibrosis. This process may be regulated by DNA methylation modification.
		                        		
		                        		
		                        		
		                        	
2.Establishment of high-throughput liquid chromatography tandem mass spectrometry method for determination of 53 per- and polyfluoroalkyl substances in serum
Zheng WANG ; Boya ZHANG ; Jiming ZHANG ; Chao FENG ; Yuanjie LIN ; Chunhua WU ; Dasheng LU ; Zhijun ZHOU
Journal of Environmental and Occupational Medicine 2024;41(4):375-383
		                        		
		                        			
		                        			Background Per- and polyfluoroalkyl substances (PFAS) are a class of persistent organic pollutants that possess potential toxicity to the human body. The production and utilization of diverse emerging PFAS have resulted in widespread human exposure. Therefore, it is imperative to establish a quantitative methodology encompassing a wide range of PFAS for a comprehensive assessment of human exposure to these compounds. Objective To establish a high-throughput quantitative method for the simultaneous determination of 53 PFAS in human serum based on ultra-high-performance liquid chromatography-Q Exactive high resolution mass spectrometry (UPLC-Q Exactive HRMS). Methods The extraction recoveries of hydrophilic-lipophilic balance (HLB) column, weak anionexchange (WAX) column, and 96-well WAX μElution plate were compared to select the SPE column with the highest recovery. The retention time and peak shape of the target compounds were compared between ACQUITY UPLC BEH C18 column and Accucore aQ column, and the more cost-effective column was chosen. The effects of adding different levels of ammonium formate (0, 2, 5 and 10 mmol·L−1) in mobile phase on peak shape and target response were compared to determine the optimal buffer salt concentration. The optimal spray voltage was obtained by comparing −2 kV and −4 kV. The proposed method was validated from the aspects of selectivity, standard curve, limits of detection, precision, accuracy, and matrix effect. The method was applied to 142 umbilical serum samples. Results The best recovery rate (64%-118%) was achieved by using 96-well WAX μElution plate. The optimal separation and peak shape were obtained by utilizing Accucore aQ column with H2O-methanol (containing 5 mmol·L−1 ammonium formate) as the mobile phase. Less in-source collision and better target response were observed when the spray voltage was set to −2 kV. All target analytes had a good linearity, with R2 > 0.99. The limits of detection ranged from 0.01 to 0.50 μg·L−1, and the recovery ranged from 69% to 127% with the precision less than 26%. A total of 31 PFAS were detected in the 142 actual samples, among which 14 PFAS had a detection frequency over 50%. Perfluorooctanoic acid showed the highest median concentration of 4.16 μg·L−1, followed by 6:2 chlorinated polyfluorinated ether sulfonate and perfluorooctane sulfonates (3.50 μg·L−1 and 1.59 μg·L−1, respectively). Conclusion In this study, we establish a UPLC-Q Excative HRMS method for simutanious determination of 53 PFAS concentrations in serum. This method has the advantages of wide coverage of PFAS, good selectivity, and easy operation, and is suitable for biological detection with a large sample size.
		                        		
		                        		
		                        		
		                        	
3.Effectss of persistent obesity on lung function in school age children
Chinese Journal of School Health 2024;45(4):549-553
		                        		
		                        			Objective:
		                        			To analyze the impact of persistent obesity on their lung function, so as to offer insights for implementing intervention measures to increase lung function in obese school age children.
		                        		
		                        			Methods:
		                        			A total of 335 children from the Sheyang Mini Birth Cohort established in 2009 in Yancheng City, Jiangsu Province, who participated in the follow up at the ages of 7 years (2016) and 10 years (2019), were selected as the study participants. Physical measurements including height, weight, and lung function were recorded. According to the World Health Organization standard, that is, gender  and age specific to correct the body mass index to calculate the body mass index  Z  score, was used to evaluate the obesity status of children at the age of 7 and 10. Children were divided into four groups, including sustained non obesity group, restored obesity group, newly classified obesity group, and persistent obesity group. Meanwhile, the lung function prediction equations recommended by the Global Lung Function Initiative were used to standardize the lung function indexes of children. Pulmonary function differences among these groups were examined, and the relationship between childhood obesity and pulmonary function was longitudinally analyzed using generalized estimating equations.
		                        		
		                        			Results:
		                        			The prevalence of obesity were 9.0% and 16.1% at the age of 7 and 10 years, respectively. The proportion of both newly classified and persistent obesity group were 8.1%, respectively. The forced expiratory volume in one second (FEV 1) and forced vital capacity (FVC) were (1 269.90±202.70) and (1 415.70±230.00) mL, respectively, at the age of 7 years. FEV 1 and FVC at the age of 10 years were (1 440.80±403.20) and (1 555.60±517.60) mL, respectively. Cross sectional analysis at age 7 showed that forced expiratory flow at 75% vital capacity (FEF 75 ) ( β=-0.52, 95%CI =-0.96--0.07) and maximal mid expiratary flow (MMEF) ( β=-0.45, 95%CI =-0.89--0.00) were significantly lower in obese children compared to their non obese peers ( P < 0.05). Longitudinal analysis indicated that obese children had lower levels of lung pulmonary function, with a statistically significant difference in FEV 1 ( β=-0.44, 95%CI=-0.85--0.02, P <0.05). There was no significant difference among the various obesity groups ( P >0.05), while gender stratified results revealed significant reductions in FEV 1/FVC in newly classified obese girls at age 10 years ( β=-1.76, 95%CI =-3.13--0.38) and in MMEF in persistently obese girls at age 10 years ( β=-1.44, 95%CI = -2.79- -0.09) ( P <0.05).
		                        		
		                        			Conclusion
		                        			Obesity may contribute to reduced lung function levels in school aged children, with newly classified and persistent obesity having more pronounced effects on lung function in girls.
		                        		
		                        		
		                        		
		                        	
4.The experience on the construction of the cluster prevention and control system for COVID-19 infection in designated hospitals during the period of "Category B infectious disease treated as Category A"
Wanjie YANG ; Xianduo LIU ; Ximo WANG ; Weiguo XU ; Lei ZHANG ; Qiang FU ; Jiming YANG ; Jing QIAN ; Fuyu ZHANG ; Li TIAN ; Wenlong ZHANG ; Yu ZHANG ; Zheng CHEN ; Shifeng SHAO ; Xiang WANG ; Li GENG ; Yi REN ; Ying WANG ; Lixia SHI ; Zhen WAN ; Yi XIE ; Yuanyuan LIU ; Weili YU ; Jing HAN ; Li LIU ; Huan ZHU ; Zijiang YU ; Hongyang LIU ; Shimei WANG
Chinese Critical Care Medicine 2024;36(2):195-201
		                        		
		                        			
		                        			The COVID-19 epidemic has spread to the whole world for three years and has had a serious impact on human life, health and economic activities. China's epidemic prevention and control has gone through the following stages: emergency unconventional stage, emergency normalization stage, and the transitional stage from the emergency normalization to the "Category B infectious disease treated as Category B" normalization, and achieved a major and decisive victory. The designated hospitals for prevention and control of COVID-19 epidemic in Tianjin has successfully completed its tasks in all stages of epidemic prevention and control, and has accumulated valuable experience. This article summarizes the experience of constructing a hospital infection prevention and control system during the "Category B infectious disease treated as Category A" period in designated hospital. The experience is summarized as the "Cluster" hospital infection prevention and control system, namely "three rings" outside, middle and inside, "three districts" of green, orange and red, "three things" before, during and after the event, "two-day pre-purification" and "two-director system", and "one zone" management. In emergency situations, we adopt a simplified version of the cluster hospital infection prevention and control system. In emergency situations, a simplified version of the "Cluster" hospital infection prevention and control system can be adopted. This system has the following characteristics: firstly, the system emphasizes the characteristics of "cluster" and the overall management of key measures to avoid any shortcomings. The second, it emphasizes the transformation of infection control concepts to maximize the safety of medical services through infection control. The third, it emphasizes the optimization of the process. The prevention and control measures should be comprehensive and focused, while also preventing excessive use. The measures emphasize the use of the least resources to achieve the best infection control effect. The fourth, it emphasizes the quality control work of infection control, pays attention to the importance of the process, and advocates the concept of "system slimming, process fattening". Fifthly, it emphasizes that the future development depends on artificial intelligence, in order to improve the quality and efficiency of prevention and control to the greatest extent. Sixth, hospitals need to strengthen continuous training and retraining. We utilize diverse training methods, including artificial intelligence, to ensure that infection control policies and procedures are simple. We have established an evaluation and feedback mechanism to ensure that medical personnel are in an emergency state at all times.
		                        		
		                        		
		                        		
		                        	
5.Levels and influencing factors of perfluorinated and polyfluoroalkyl substances in umbilical cord serum from Sheyang Mini Birth Cohort Study, Jiangsu Province
Ruonan TAN ; Zheng WANG ; Jiming ZHANG ; Yiming DAI ; Jianqiu GUO ; Xiaojuan QI ; Dasheng LU ; Xiuli CHANG ; Chunhua WU ; Zhijun ZHOU
Journal of Environmental and Occupational Medicine 2024;41(8):841-848
		                        		
		                        			
		                        			Background Perfluorinated and polyfluoroalkyl substances (PFAS), a large group of emerging pollutants, are ubiquitous in the ecological environment. Their multiple organ toxic effects on human body are reported. Understanding the exposure level of PFAS in cord serum and associated influencing factors can provide scientific evidence for studying maternal and newborn health effects and risk regulation. Objective To explore the exposure levels of PFAS in cord serum and potential impact factors. Methods This study was based on the maternal and infant database and the cord serum sample bank of the Sheyang Mini Birth Cohort Study (SMBCS) established in 2009. A self-designed questionnaire was used to collect information on sociodemographic characteristics, living environment, and lifestyle of mothers during pregnancy. A total of 
		                        		
		                        	
6.Complete androgen insensitivity syndrome with gender transition in adulthood: A case report
Meicen PU ; Dan WANG ; Meinan HE ; Xinzhao FAN ; Mengchen ZOU ; Yijuan HUANG ; Jiming LI ; Shanchao ZHAO ; Yunjun LIAO ; Yaoming XUE ; Ying CAO
Chinese Journal of Endocrinology and Metabolism 2024;40(7):602-607
		                        		
		                        			
		                        			Complete androgen insensitivity syndrome(CAIS) is characterized by lack of androgen response in target organs due to androgen receptor dysfunction, resulting in feminized external genitalia. Individuals with CAIS are typically advised to live as females. This article reports a patient diagnosed with CAIS and gender dysphoria in adulthood. Following the removal of a left pelvic mass, pathology indicated cryptorchidism with a concurrent Leydig cell tumor. Genetic testing revealed a deletion mutation in exon 3 of androgen receptor gene. During follow-up, the patient underwent gender reassignment, transitioning socially from female to male. This case provides new insights into gender allocation for CAIS patients.
		                        		
		                        		
		                        		
		                        	
7.Analysis of the changes in management and allocation planning of large medical equipment in China
Wei XIONG ; Jiming WANG ; Tao BAO ; Jing SUN ; Ge CHEN ; Jingyi FENG
Chinese Journal of Hospital Administration 2024;40(7):547-553
		                        		
		                        			
		                        			Objective:To analyze the changes in the management methods of large medical equipment in China, evaluate the equity of large medical equipment allocation planning from the 13th Five-Year Plan to the 14th Five-Year Plan.Methods:The literature research was used to sort out the characteristics and changes of the management of large medical equipment in China. The database was established to quantitatively analyze the existing number of large medical equipment at the end of the 13th Five-Year Plan and the planned number of large medical equipment at the end of the 14th Five-Year Plan, and then the fairness evaluation was carried out based on the Lorenz curve and Gini coefficient.Results:Since the 1980s, the management measures of large-scale medical equipment in China have been introduced and updated iteratively to meet the needs of social development and medical services. In addition to conventional radiotherapy equipment, the number of large medical equipment per million population in China at the end of the 14th Five-Year Plan at least doubled compared with the end of the 13th Five-Year Plan, among which the heavy ion radiotherapy system and the laparoscopic surgery system increased by more than two times. Lorenz curve analysis showed that the fairness of large medical equipment based on population distribution at the end of the 14th Five-Year Plan was better than that at the end of the 13th Five-Year Plan, especially for heavy ion proton radiotherapy system, high-end radiotherapy equipment and PET/MR. The Gini coefficient of large medical equipment planning based on population distribution during the 14th Five-Year Plan was generally smaller than that of the 13th Five-Year Plan. Except for the heavy ion proton radiotherapy system and PET/MR in North China, the Gini coefficient of the planned number of all kinds of equipment in the six regions of the country during the 14th Five-Year Plan was less than 0.4 based on population distribution, which was in a relatively fair state.Conclusions:The management of large medical equipment in China keeps pace with the times and constantly adapts to the new development stage of health care. The fairness of large medical equipment allocation planning based on population distribution in six regions of China has been steadily improved, and it is suggested that the fairness based on population distribution and the clinical application of existing equipment should be taken into account in the allocation planning.
		                        		
		                        		
		                        		
		                        	
8.Analysis on the characteristics and liver function of occupational hepatotoxicants exposed population
Jie WANG ; Zhiping DUAN ; Yubo LIU ; Jiming ZHANG ; Zhijun ZHOU ; Xuetao ZHANG
China Occupational Medicine 2023;50(3):312-316
		                        		
		                        			
		                        			Objective  To investigate the characteristics and liver function of the population with occupational exposure to hepatotoxicants. Methods  A total of 17 093 workers with occupational hepatotoxicants exposure who underwent occupational medical examination during their employment in a occupational medical examination institution of Shanghai in 2021 were selected as the research subjects by judgement sampling method. Occupational medical examination data were collected, and the prevalence of abnormal liver function and fatty liver were analyzed. The association between hepatotoxicants exposure and abnormal liver function were analyzed. Results  The median and the 0th-100th percentiles of the duration of exposure to hepatotoxicants was 6.5(1.0-42.0) years. The prevalence of fatty liver was 48.4% and the incidence of abnormal liver function was 23.7%. Among the workers with fatty liver, the prevalence of abnormal liver function was higher in workers exposed to metals, metalloids and their compounds than in unexposed workers (33.9% vs 30.0%, P<0.01). The results of multivariate logistic regression analysis demonstrated that the risk of abnormal liver function increased with the number of different hepatotoxicants mixed exposures (all P<0.01), after correcting for confounding factors including gender, age, years of exposure, marital status, drinking, hypertension, fatty liver and blood sugar. Conclusion  Exposure to hepatotoxicants is a risk factor for abnormal liver function. The more diverse types of hepatotoxicants an individual is exposed to, the stronger the association with this risk. 
		                        		
		                        		
		                        		
		                        	
9.Epidemiological characteristics of foodborne disease outbreaks in Shaoxing City from 2012 to 2022
XING Chao ; WANG Qimei ; REN Jianglei ; CHEN Jiming ; HE Qinfen ; JIANG Zhuojing
Journal of Preventive Medicine 2023;35(6):506-508,513
		                        		
		                        			Objective:
		                        			To investigate the epidemiological characteristics of foodborne disease outbreaks in Shaoxing City, Zhejiang Province, from 2012 to 2022, so as to provide the evidence for improving the foodborne disease control strategy. 
		                        		
		                        			Methods:
		                        			Foodborne disease outbreaks in Shaoxing City from 2012 to 2022 were collected from National Foodborne Disease Outbreak Monitoring System in China, including populations, places of outbreak, pathogenic factors and suspected foods. The temporal distribution, regional distribution, distribution of outbreak places and pathogenic factors of foodborne disease outbreaks were descriptively analyzed.
		                        		
		                        			Results:
		                        			A total of 89 foodborne disease outbreaks were reported in Shaoxing City from 2012 to 2022, covering totally 699 patients, with an average annual attack rate of 6.35%. The outbreak peaked during the period between June and October (73 outbreaks, 82.02%), and family was the predominant place of outbreak (41 outbreaks, 46.07%). There were 83 outbreaks with known pathogenic factors, including 51 outbreaks caused by microbial factors, with Vibrio parahaemolyticus, Salmonella and norovirus as predominant pathogens, and 29 outbreaks caused by fungi and their toxins, which were all poisonous mushrooms poisoning, resulting in 2 deaths. In addition, there were 3 outbreaks caused by chemical factors. 
		                        		
		                        			Conclusions
		                        			The outbreak of foodborne diseases predominantly occurred in summer and autumn in Shaoxing City from 2012 to 2022. Family was the predominant place of outbreak, and toxic mushroom poisoning was the most lethal pathogenic factor.
		                        		
		                        		
		                        		
		                        	
10.Effects of flurochloridone on apoptosis and Nrf2/HO-1 and NFκB signaling pathways in mouse testis and TM4 cells
Shuqi ZHAO ; Yanna WANG ; Zhijing NI ; Fang TIAN ; Rui LI ; Jiming ZHANG ; Xiuli CHANG ; Zhijun ZHOU
Journal of Environmental and Occupational Medicine 2023;40(6):673-680
		                        		
		                        			
		                        			Background Flurochloridone (FLC) is toxic to male reproduction and can induce apoptosis of testicular tissue and supporting cells under oxidative stress. Of particular concern is whether nuclear factor-erythrocyte 2-related factor 2/heme oxygenase-1 (Nrf2/HO-1) signaling pathway and nuclear factor kappa B (NFκB) signaling pathway participate this process. Objective To observe apoptosis of testicular tissue and sertoli TM4 cells and alterations of Nrf2/HO-1 and NFκB signaling pathways in mice treated with FLC in vivo/in vitro. Methods (1) Animal experiment. Testis samples were harvested from male C57BL/6 mice after 28-day FLC (0, 3, 15, 75, and 375 mg·kg−1 per day) exposure via oral route. Malondialdehyde (MDA) and superoxide dismutase (SOD) in homogenate of testicular tissue were measured by colorimetry. Apoptosis of testicular tissue was evaluated by TUNEL staining. Expression and distribution of Nrf2 and NFκB were detected by immunohistochemistry. Protein expression levels of Nrf2, HO-1, NAD(P)H: quinone oxidoreductase 1 (NQO1), NFκB, inhibitor of nuclear factor kappa-B kinase subunit beta (IKKβ), and phosphorylated recombinant inhibitory subunit of nuclear factor kappa-B alpha (P-IκBα) in testicular tissue homogenate were determined by Western blotting. (2) Cell experiment. TM4 cell lines were treated with 40, 80, 120, 160, and 200 μmol·L−1 FLC for 6 h, and cell viability was detected by CCK-8. After 6 h exposure to 40, 80, and 160 μmol·L−1 FLC, the apoptosis rate was detected by flow cytometry, and the protein expression levels of Nrf2, HO-1, NQO1, NFκB, IKKβ, and IκBα were detected by Western blotting. Results (1) Animal experiment. Apoptosis occurred in the interstitial and basal parts of spermatogenic tubules in male C57BL/6 mice after 28 days of oral FLC exposure. Compared with the control group, the MDA level in testicular tissue of the 375 mg·kg−1 FLC-treated group was significantly increased (P<0.05), and the SOD activity was significantly decreased (P<0.05). After 375 mg·kg−1 FLC exposure, apoptosis occurred in the interstitial and basal parts of spermatogenic tubules. The results of immunohistochemistry showed the expression of Nrf2 and NFκB in the interstitium and basal part of spermatogenic tubules of the treated groups. Compared with the control group, the protein levels of Nrf2, NQO1, P-IκBα, NFκB, and IKKβ in the 15, 75, and 375 mg·kg-1 groups were significantly increased (P<0.001), and the HO-1 protein level was significantly increased in the 375 mg·kg−1 group (P<0.001). (2) Cell experiment. Compared with the control group, the TM4 cell viabilities in the 40, 80, 120, 160, and 200 μmol·L−1 FLC-treated groups significantly decreased (P<0.01). The apoptosis rates were significantly increased (P<0.05), and the apoptosis rates increased from 5.7% in the control group to 7.4%, 9.4%, and 11.7% in the 40, 80, and 160 μmol·L−1, respectively. The Nrf2 protein level in the 40 μmol·L−1 group was significantly increased (P<0.01), while the levels significantly decreased in the 80 and 160 μmol·L−1 groups (P<0.01). The HO-1 protein levels in the 40, 80, and 160 μmol·L−1 groups were significantly increased (P<0.01). The level of NQO1 protein in the 40 μmol·L−1 group was significantly increased (P<0.01). The NFκB protein levels were significantly increased in the 80 and 160 μmol·L−1 groups (P<0.001). The IκBα protein levels were significantly decreased in all treated groups (P<0.001). The IKKβ protein had no significant change. Conclusion FLC induces testicular tissue apoptosis, and the process affects Nrf2/HO-1 signaling pathway and NFκB signaling pathway. The in vitro study confirms that FLC could induce apoptosis of TM4 cells and activate Nrf2/HO-1 and NFκB signaling pathways.
		                        		
		                        		
		                        		
		                        	
            

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