1.Rapid culture and identification of mouse primary thyroid cells
Qiuchan TAN ; Jiawei LIN ; Xiaoya YANG ; Li PAN ; Dandan YAO ; Liwei WANG ; Lixin CHEN ; Zhuhua WU
Chinese Journal of Pathophysiology 2024;40(3):572-576
AIM:The paper is to explore a rapid and simple method for the culture of mouse primary thyroid cells.METHODS:Mouse thyroid cells were isolated by enzyme digestion and cultured with improved medium,and their morphology,characteristics and secretory function were observed within 14 d.RESULTS:In the cultures,the active pri-mary cells were obtained from the thyroid tissue after digestion for 25 min;adherent growth was observed on the 2nd day.And secondary follicles appeared from the 5th to 7th day.Over 95%cells were detected with thyroglobulin.The secretion of total triiodothyronine and total thyroxine maintains over 60%in 7 d.The expression levels of specific genes can still maintain more than 50%in 10 d.CONCLUSION:Mouse thyroid primary cells can be rapidly cultured by this method,and the cells can be used for studying thyroid endocrine secretion within 7 d and studying thyroid genes within 10 d.
2.Prevalence and genetic characteristics of Cryptosporidium infections among HIV-positive individuals in Jiangxi Province
Zhuhua HU ; Liang LU ; Yingfang YU ; Lin LI ; Wei WANG ; Guoyin FAN ; Changhua FENG ; Yangyun ZHENG ; Guohua PENG
Chinese Journal of Schistosomiasis Control 2024;36(6):637-642
Objective To investigate the prevalence of Cryptosporidium infection and the distribution of parasite species and genotypes among HIV-positive individuals in Jiangxi Province. Methods HIV-positive individuals' sociodemographic and clinical data were collected from three AIDS designated hospitals in Jiangxi Province from January 2022 to March 2023. Subjects' stool samples were collected, and genomic DNA was extracted from stool samples. Nested PCR assay was performed based on the small subunit ribosomal RNA (SSU rRNA) gene of Cryptosporidium, and Cryptosporidium gp60 gene was amplified in stool samples positive for the SSU rRNA gene. The second-round PCR amplification product was checked with 1.5% agarose gel electrophoresis, and the products of suspected positive amplifications were sequenced, followed by sequence alignment. The phylogenetic tree was created using the Neighbor-Joining method with the software MEGA 11.0, to characterize the species, genotypes and sub-genotypes of Cryptosporidium. Results A total of 382 HIV-positive individuals were enrolled, with two cases identified with Cryptosporidium infection (0.52% prevalence), and both cases had no abdominal pain or diarrhea. Following sequencing and sequence alignment, the gene sequences of these two Cryptosporidium isolates shared 99.76% and 99.88% similarity with the gene sequence of C. meleagridis isolates. Phylogenetic analysis based on the Cryptosporidium SSU rRNA gene sequence identified the species of these two Cryptosporidium-positive stool samples as C. meleagridis. Following nested PCR amplification of the Cryptosporidium gp60 gene, sequencing and sequence alignment, the two C. meleagridis isolates were characterized as III eA17G2R1 and III bA25G1R1a sub-genotypes, and the sub-genotype III bA25G1R1a was firstly described in humans. Conclusion The prevalence of Cryptosporidium is low among HIV-positive individuals in Jiangxi Province. The likelihood of Cryptosporidium infection cannot be neglected among HIV-positive individuals without diarrhea.
3.IDI2-AS1 influences the development of acute myocardial infarction by regulating NR4A2 through microRNA-33b-5p
Shuxing WU ; Zhihua PANG ; Ru WANG ; Jian CUI ; Wenting LI ; Xiaoyu YANG ; Zhuhua YAO
Chinese Critical Care Medicine 2024;36(9):972-979
Objective:To explore the effect and correlation of long non-coding RNA (lncRNA) IDI2-AS1/microRNA-33b-5p (miR-33b-5p)/nuclear receptor-associated protein NR4A2 competitive endogenous RNA (ceRNA) regulatory network on acute myocardial infarction (AMI), and to verify whether IDI2-AS1 regulates NR4A2 through miR-33b-5p to affect the occurrence and development of myocardial infarction.Methods:The miRNA and mRNA expression chips related to myocardial infarction were obtained from gene expression omnibus (GEO), and the differential expression was analyzed. The upstream regulatory mechanism of NR4A2 was predicted using TargetScan database. Thirty-two male C57/BL6 mice were divided into Sham group, AMI model group, miR-33b-5p mimic group [miR-33b-5p mimic lentivirus (5×10 7 TU) was injected locally into the heart tissue during ligation] and miR-33b-5p inhibitor group [miR-33b-5p inhibitor lentivirus (5×10 7 TU) was injected locally into the heart tissue during ligation] according to random number table method, with 8 mice per group. Left ventricular end-diastolic diameter (LVEDD) and left ventricular end-systolic diameter (LVESD) were asseessed by echocardiography, left ventricular fractional shortening (LVFS) and left ventricular ejection fraction (LVEF) were calculated. After the last weighing, the anesthetized mice were sacrificed and the heart tissues were taken. Masson staining of the heart tissues was observed under light microscope, myocardial collagen volume fraction (CVF) and infarct size were calculated. Cardiomyocytes of SPF grade SD rats were collected. They were divided into normal control group (control group), ischemia-hypoxia model group, miR-33b-5p mimic transfection group (miR-33b-5p mimic transfection group before ischemia and hypoxia treatment) and miR-33b-5p inhibitor transfection group (miR-33b-5p inhibitor transfection group before ischemia and hypoxia treatment). The activity of caspase-3/7 in cardiomyocytes was measured. The levels of interleukins (IL-1β, IL-6) and tumor necrosis factor-α (TNF-α) were detected by enzyme-linked immunosorbent assay (ELISA). The levels of malondialdehyde (MDA), superoxide dismutase (SOD), creatine kinase (CK), MB isoenzyme of creatine kinase (CK-MB) and lactate dehydrogenase (LDH) were detected by colorimetry. Real-time quantitative polymerase chain reaction (RT-qPCR) was used to detect the expression of apoptosis-related proteins Bax and Bcl-2, cytochrome C (Cyt C) and IDI2-AS1/miR-33b-5p/NR4A2 regulatory axis genes. Results:The myocardial infarction microarray analysis showed that NR4A2 expression was significantly up-regulated in myocardial infarction, with predicted upstream regulatory mechanisms indicating its possible influence through the IDI2-AS1/miR-33b-5p/NR4A2 regulatory axis. Echocardiographic detection showed that compared with AMI model group and miR-33b-5p inhibitor group, LVEF and LVFS in the heart tissue of mice in miR-33b-5p mimic group were significantly increased, while the levels of LVEDD, LVESD, CK, CK-MB and LDH were significantly decreased, with statistical significance. Light microscope showed myocardial fibrosis and myocardial infarction in AMI model group and miR-33b-5p inhibitor group. In the miR-33b-5p mimic group, the degree of myocardial fibrosis was decreased and the myocardial infarction size was significantly reduced. Compared with AMI model group and miR-33b-5p inhibitor group, the levels of MDA, IL-1β, IL-6, TNF-α and the expressions of Bax and Cyt C in the heart tissue of mice in miR-33b-5p mimic group were significantly decreased, while the levels of SOD and Bcl-2 expression were significantly increased, and the differences were statistically significant. The expressions of IDI2-AS1 and NR4A2 in the heart tissue of mice in miR-33b-5p mimic group were significantly lower than those in AMI model group and miR-33b-5p inhibitor group [IDI2-AS1 (2 -ΔΔCt): 1.96±0.08 vs. 2.73±0.08, 3.10±0.05, NR4A2 (2 -ΔΔCt): 2.36±0.07 vs. 3.16±0.08, 3.80±0.08, all P < 0.01]. The expression of miR-33b-5p was significantly higher than that of AMI model group and miR-33b-5p inhibitor group (2 -ΔΔCt: 0.88±0.07 vs. 0.57±0.07, 0.23±0.01, both P < 0.01). The cell experiment results showed that the caspase-3/7 activity of rat neonatal cardiomyocytes in the miR-33b-5p mimic transfection group was significantly lower than that in the ischemia-hypoxia model group and the miR-33b-5p inhibitor transfection group, suggesting that miR-33b-5p can significantly reduce the apoptosis level of the ischemia-hypoxia model. The levels of peroxidation and inflammation indexes, important genes of apoptosis pathway and the expression of IDI2-AS1/miR-33b-5p/NR4A2 regulatory axis of rat neonatal cardiomyocytes in all groups were consistent with the above. Conclusion:IDI2-AS1 can regulate NR4A2 through miR-33b-5p, thus affecting the occurrence and development of AMI.
4.Analysis of chest imaging features of novel coronavirus pneumonia, bacterial pneumonia and viral pneumonia
Yufang CAO ; Xiaozhi WANG ; Xiaohong XIE ; Jinghui LI ; Chao DENG ; Xiangying LI ; Zhuhua ZHU ; Zhidian WU ; Chao JI ; Yi NIU ; Fan LIU ; Yanmei YU ; Wei SONG
Chinese Critical Care Medicine 2023;35(1):28-31
Objective:To investigate and summarize the chest CT imaging features of patients with novel coronavirus pneumonia (COVID-19), bacterial pneumonia and other viral pneumonia.Methods:Chest CT data of 102 patients with pulmonary infection due to different etiologies were retrospectively analyzed, including 36 patients with COVID-19 admitted to Hainan Provincial People's Hospital and the Second Affiliated Hospital of Hainan Medical University from December 2019 to March 2020, 16 patients with other viral pneumonia admitted to Hainan Provincial People's Hospital from January 2018 to February 2020, and 50 patients with bacterial pneumonia admitted to Haikou Affiliated Hospital of Central South University Xiangya School of Medicine from April 2018 to May 2020. Two senior radiologists and two senior intensive care physicians were participated to evaluated the extent of lesions involvement and imaging features of the first chest CT after the onset of the disease.Results:Bilateral pulmonary lesions were more common in patients with COVID-19 and other viral pneumonia, and the incidence was significantly higher than that of bacterial pneumonia (91.6%, 75.0% vs. 26.0%, P < 0.05). Compared with other viral pneumonia and COVID-19, bacterial pneumonia was mainly characterized by single-lung and multi-lobed lesion (62.0% vs. 18.8%, 5.6%, P < 0.05), accompanied by pleural effusion and lymph node enlargement. The proportion of ground-glass opacity in the lung tissues of patients with COVID-19 was 97.2%, that of patients with other viral pneumonia was 56.2%, and that of patients with bacterial pneumonia was only 2.0% ( P < 0.05). The incidence rate of lung tissue consolidation (25.0%, 12.5%), air bronchial sign (13.9%, 6.2%) and pleural effusion (16.7%, 37.5%) in patients with COVID-19 and other viral pneumonia were significantly lower than those in patients with bacterial pneumonia (62.0%, 32.0%, 60.0%, all P < 0.05), paving stone sign (22.2%, 37.5%), fine mesh sign (38.9%, 31.2%), halo sign(11.1%, 25.0%), ground-glass opacity with interlobular septal thickening (30.6%, 37.5%), bilateral patchy pattern/rope shadow (80.6%, 50.0%) etc. were significantly higher than those of bacterial pneumonia (2.0%, 4.0%, 2.0%, 0%, 22.0%, all P < 0.05). The incidence of local patchy shadow in patients with COVID-19 was only 8.3%, significantly lower than that in patients with other viral pneumonia and bacterial pneumonia (8.3% vs. 68.8%, 50.0%, P < 0.05). There was no significant difference in the incidence of peripheral vascular shadow thickening in patients with COVID-19, other viral pneumonia and bacterial pneumonia (27.8%, 12.5%, 30.0%, P > 0.05). Conclusions:The probability of ground-glass opacity, paving stone and grid shadow in chest CT of patients with COVID-19 was significantly higher than those of bacterial pneumonia, and it was more common in the lower lungs and lateral dorsal segment. In other patients with viral pneumonia, ground-glass opacity was distributed in both upper and lower lungs. Bacterial pneumonia is usually characterized by single lung consolidation, distributed in lobules or large lobes and accompanied by pleural effusion.
5.Construction of evaluation index system of peritoneal dialysis nursing quality based on Delphi method and analytic hierarchy process
Zhuhua ZHAO ; Faxian DING ; Xiuping YIN ; Zhihui WANG ; Weihua ZHAO ; Lixia WAN ; Li MA ; Tingxin WAN
Chinese Journal of Modern Nursing 2021;27(15):2054-2060
Objective:To construct the evaluation index system of peritoneal dialysis nursing quality, so as to provide a basis for management and evaluation of peritoneal dialysis nursing quality.Methods:Through literature review and expert interviews, the peritoneal dialysis nursing quality evaluation index system (draft) was initially determined on the evidence-based basis. The expert letter questionnaire was designed, and indexes of nursing quality evaluation were screened and revised by Delphi expert letter questionnaire and boundary value method. The analytic hierarchy process was used to determine the weight of evaluation indexes, and the evaluation system of the peritoneal dialysis nursing quality index was established.Results:The expert response rates of 2 rounds of questionnaires were 91.43% and 85.71%, and the expert authority coefficients of the 2 rounds were respectively 0.838 0 and 0.862 2, and the coordination coefficients of the first, second and third levels of indexes of the second round were respectively 0.643, 0.431 and 0.283 ( P<0.01) . The final evaluation index system included 3 first-level indicators, 18 second-level indicators and 84 third-level indicators. Conclusions:The evaluation index system of peritoneal dialysis nursing quality has good reliability and applicability. After further verification, it can be used as an evaluation tool of peritoneal dialysis nursing quality management.
6. The relationship between openness and social adaptation among offenders in community: the mediating role of interpersonal sensitivity and the moderating role of peer trust
Zhuhua LIU ; Jie XU ; Yi LI ; Wei GUO
Chinese Journal of Behavioral Medicine and Brain Science 2019;28(12):1131-1135
Objective:
To explore the mediating effect of interpersonal sensitivity between social adaptation and openness, as well as the moderating role of peer attachment among offenders in community.
Methods:
Totally 155 offenders from 5 communities in Yinchuan city were investigated by symptom self-rating scale, NEO five-factor inventory(NEO-FFI), social adaptability scale and the inventory of parent and peer attachment-revised. Using the macro program PROCESS of SSPS to test the mediating effect of interpersonal sensitivity and the moderating role of peer trust.
Results:
The scores of interpersonal sensitive subscale of the symptom checklist 90, the openness subscale of NEO five-factor inventory, the inventory of parent and peer attachment-revised were (1.52±0.48), (3.05±0.30), (0.72±0.60) and (83.40±12.17), respectively. Openness, peer attachment were positively correlated with social adaptation (
7.Third-generation Dual-source CT for Head and Neck CT Angiography with 70 kV Tube Voltage and 20-25 ml Contrast Medium in Patients With Body Weight Lower than 75 kg.
Yu CHEN ; Yuanli ZHU ; Huadan XUE ; Yun WANG ; Yumei LI ; Zhuhua ZHANG ; Zhengyu JIN
Acta Academiae Medicinae Sinicae 2017;39(1):4-8
Objective To investigate the image quality of head and neck CT angiography (CTA)using the third-generation dual-source CT combined with 70 kV tube voltage and 20-25 ml contrast medium (CM),and evaluate the effects of venous artifacts arising from the CM on the ipsilateral side of injection. Methods Totally 40 consecutive patients with suspected vascular diseases and body weight lower than 75 kg prospectively underwent head and neck CTA examination using the third-generation dual-source CT. CTA was performed with a third-generation dual-source CT system. Patients were randomly divived into 70 kV group (n=20)and 100 kV group (n=20). The 70 kV group used 20-25 ml CM and advanced modeled iterative reconstruction technique,and the 100 kV group used 40 ml CM and filtered back projection. Venous artifacts and CM residues were evaluated by a 3-point scale (1=excellent,3=poor),respectively. Results The effective dose of 70 kV group decreased 58% compared to 100 kV group (t=-18.14,P<0.001).In the 70 kV group,16 patients (80.0%)presented with venous artifacts and six of them (37.5%,6/16)affected the adjacent arteries. In the 100 kV group,19 patients (95.0%)presented with venous artifacts,and seven of them (36.8%,7/19)affected the adjacent arteries (Z=-0.878,P=0.380). In the 70 kV group,13 patients (65.0%)presented with obvious CM residues and two of them (15.3%,2/13)prolonged into the superior vena cava (SVC). In the 100 kV group,19 patients(95.0%)presented with obvious CM residues,and thirteen of them(68.4%,13/19)prolonged into the SVC (Z=-3.654,P<0.001). Conclusion Compared with the 100 kV,the third-generation dual-source CT for head and neck CTA,combined with 70 kV and 20-25 ml CM,can remarkably decrease the radiation dose,along with reduced CM residues and comparable venous artifacts.
Artifacts
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Body Weight
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Computed Tomography Angiography
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methods
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Contrast Media
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Head
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diagnostic imaging
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Humans
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diagnostic imaging
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Radiation Dosage
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Radiographic Image Interpretation, Computer-Assisted
8. Laryngeal endoscopic cricopharyngeal myotomy for cricopharyngeal achalasia post stroke
Jian WANG ; Wuyi LI ; Yongjin LI ; Dahai YANG ; Hong HUO ; Xiaofeng JIN ; Yanyan NIU ; Xu TIAN ; Zhuhua ZHANG ; Yu CHEN ; Zhiqiang GAO
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2017;52(10):729-732
Objective:
To explore the safety and validity of endoscopic cricopharyngeal myotomy in patients with cricopharyngeal achalasia.
Methods:
A total of 19 patients with cricopharyngeal achalasia suffered from sustained dysphagia were enrolled in this study. The patients were divided into transcervical cricopharyngeal myotomy(CPM) group and endoscopic CPM (ECPM) group. Swallowing function and complications were evaluated.SPSS7.0 software was used to analyze the data.
Results:
The swallowing function improved significantly in seven patients in ECPM group, and 9 patients improved in CPM group.The video fluoroscopic swallowing study(VFSS)-swallowing score, VFSS-aspiration score and drinking test score were (3.1±1.1), (3.4±0.8) and (2.0±0.6)in post-ECPM, (3.4±1.4), (3.0±0.9) and (2.2±0.6)in post-CPM. No statistical difference was found in validity between CPM group and ECPM group(
9.Feasibility study of guiding catheter passing through spasmodic vess els during percutaneous coronary intervention via radial artery access by the aid of PCI guiding wire and balloon
Zhuhua NI ; Lefeng WANG ; Xinchun YANG ; Hongshi WANG ; Li XU ; Weiming LI ; Kun XIA ; Yu LIU ; Jifang HE ; Yonghui CHI ; Dapeng ZHANG ; Junping DENG ; Yimin WANG ; Guangjun LIU ; Xiaoliang ZHANG ; Jianhong ZHAO ; Jiqiang ZHANG ; Jiasheng LIU ; Shuying QI
Chinese Journal of Interventional Cardiology 2016;24(6):320-325
Objective To explore the safety and feasibility of guiding catheter passing through spasmodic vessels in patients undergoing percutaneous coronary intervention (PCI) via radial artery access by the aid of PCI guiding wire and balloon .Methods The clinical data of 33 coronary artery disease (CAD) patients undergoing PCI via radial artery access with radial artery or (and) brachial artery spasm ( group A ) were retrospectively analyzed .Among all these patients , guiding catheters were delivered through the spasmodic vessels successfully by the aid of PCI guiding wires and balloons .The clinical data of other 38 CAD patients having PCI during the same period performed by other operators via radial artery or ( and ) brachial artery approach and experienced vessel spasm were anlysed as the control ( group B ) .All patients in group B received conventional anti-spasm management during PCI .All vessel spasm was identified by angiography.For patients in group A , a diameter of 0.014 inch guiding wire was chosen to pass through the spasmodic vessel segment carefully and gently .The diameter of balloon should be chosen according to the diameter of guiding catheter .A balloon diameter of 2.0 mm and 2.5 mm was corresponded to 6F and 7F guiding catheter respectively .The balloon was advanced to the tip of guiding catheter , keeping a half in catheter and a half in vessel followed by inflating the balloon with a pressure of 8 atm.The balloon was kept inflated the guiding catheter was pushed in vitro carefully and slowly until the catheter passed through the spasmodic vessel segment .Then the balloon was deflated and pulled out together with PCI guiding wire . Exchanged a diameter of 0.035 inch wire and completed the positioning of guiding catheter .After finishing the PCI, radial or ( and) brachial angiography was performed again to observe if spasm disappeared and to determine if there any contrast medium exudation .For patients in group B , routine approach was applied including administration of nitroglycerine , diltiazem or nitroprusside etc . to relieve vessel spasm. Results The location of vessel spasm was similar in group A and group B ( P=0.150 ) , and the incidence rate of spasm in brachial artery was higher than that in radial artery in both groups .The chance of guiding catheter crossing the spasmodic vessel segment was significantly higher in group A than in group B ( 100%vs.39.5%, P=0.00).In patients whose guiding catheter could pass through the spasmodic vessel segment successfully , time spent in group A was shorter than in group B ( P=0.000 ) .The patient number which time spent was less than five minutes , five to 15 minutes and more than 15 minutes was 30 and 2 ( 90.1%vs.13.3%) , 3 and 7 ( 9.9% vs.46.7%) and 0 and 6 ( 0% vs.40.0%) in group A and in group B respectively.The incidence of forearm hematoma was lower in group A than in group B without statistical difference [6.1%(2/33) vs.18.4%(7/38), P =0.113].Conclusions It is safe and feasible for passing guiding catheter through spasmodic vessels during PCI via radial artery access by the aid of PCI guiding wire and balloon .
10.Analysis of urinary iodine monitoring results of school children aged from 8 to 10 in Nanchang City, Jiangxi Province
Renlong FU ; Ming LI ; Guohua PENG ; Weihua HUANG ; Jun LING ; Zhuhua HU ; Xiaowu FENG
Chinese Journal of Endemiology 2016;35(12):892-895
Objective To survey the urinary iodine (UI) status of school children aged from 8 to 10 in Nanchang City, and to provide a scientific basis for preventing and controlling the iodine deficiency disorders (IDD). Methods From 2009 to 2012, Donghu, Xihu, Qingshanhu, Qingyunpu, Wanli, Nanchang, Xinjian, Jinxian and Anyi 9 counties (areas) were chosen in Nanchang City as monitoring areas, and five townships were selected according to the five directions as east, west, south, north and centre in each county (area), one school was selected in each township, 20 school children aged from 8 to 10 (10 males and 10 females) were chosen as respondents. Ammonium persulfate digestion-arsenic cerium catalytic spectrophotometry (WS/T 107-2006) was used to detect UI. The monitoring data on UI of 8 to 10 years old school-age children were collected and analyzed. Results From 2009 to 2012, a total of 3 600 urine samples were collected, the median of urinary iodine (MUI) was 257.35 μg/L. In the 4 years, the MUI of school children aged from 8 to 10 was 315.30, 314.80, 262.92 and 112.73 μg/L, respectively, the MUI decreased year by year, the difference was statistically significant (χ2=631.129, P<0.05). Compared with 2009 and 2010, the proportion [15.22% (137/900), 14.67% (132/900), 25.11% (226/900), 30.22%(272/900)] of MUI of 100 -199μg/L (moderate intake) in 2011 and 2012 increased year by year; the difference was statistically significant (χ2=93.977, P< 0.05). The MUIs between different counties (areas) were statistically significant (χ2=36.520, P<0.05). The MUI of children aged 8 (280.10 μg/L), 9 (255.11 μg/L) and 10 (249.20 μg/L) decreased with increasingage (χ2 = 7.813, P < 0.05). The MUI of male students (269.70 μg/L) was higher than that of female students (247.60μg/L), the difference was statistically significant (Z = - 3.704, P < 0.01). Conclusions Iodine nutrition status of 8 to 10 years old school-age children in Nanchang City is good, iodine intake meets the body's need. It is suggested that the monitoring for UI of school-age children should be strengthened in order to prevent the potential harmful effects of inappropriate iodine intake.

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