1.Differences in deltamethrin resistance and kdr gene mutation in Culex tritaeniorhynchus population in and outside the Yellow Sea wetland
Xiao-er ZHANG ; Zhi-ming WU ; Ye TIAN ; Qian CUI ; Yu-qian JI ; Huan WANG ; Shu-juan YANG ; Yi-chao ZHAO ; Yu WANG ; Hua-yu YIN ; Yu DING ; Guo-jin YAN ; Min-sen ZHAO ; Shou-gang ZHANG ; Bing-dong SONG ; Hong-na CHEN ; Jian GAO ; Wei-fang YANG ; Yu-fu ZHANG ; Hui LIU ; Hong-liang CHU
Acta Parasitologica et Medica Entomologica Sinica 2026;33(2):101-107
Objective To gain insights into the biological characteristics of different populations of Culex tritaeniorhynchus within and around the Yellow Sea wetland from the perspective of the occurrence of resistance, we investigated the levels of resistance to deltamethrin and kdr gene mutation in the wetland and its peripheral areas. Methods Specimens were collected from Cx. tritaeniorhynchus populations at two monitoring sites in the Rare Bird National Nature Reserve and Tiaozi Ni Wetland Scenic Area, and also from two populations in Yancheng City and the Liuhe District of Nanjing, and the resistance of these mosquitoes to deltamethrin was determined using the CDC biotest bottle method. For each concentration of deltamethrin assessed, a random subset of exposed specimens was selected for amplification of the kdr gene fragment, followed by Sanger sequencing to identify and analyze resistance-associated mutations. Results The LC50 levels of deltamethrin among mosquitoes from the four populations in Luhe, Yancheng, the Rare Bird National Nature Reserve and the Tiaozi Ni Wetland Scenic Area were 2.048 5, 7.798 2, 3.473 3, and 17.695 5 mg/mL, respectively, with corresponding concentrations of deltamethrin ranging from 0.005 to 5.000,0.050 to 50.000,0.050 to 25.000 and 0.050 to 50.000 mg/mL, respectively. Furthermore, the ranges of the KT50 values were 11.76-107.43, 67.05-216.30,29.77-107.43 and 28.40-329.51 min; the 1-h knockdown rates were 34.58%-99.15%, 9.52%-43.80%, 55.09%-73.01%, and 10.09%-68.07%; and the 24-h mortality rates were 12.15%-67.52%,9.52%-79.56%,13.17%-82.21%, and 11.01%-78.99%, respectively. With respect to kdr gene mutation, we assayed a total of 63,70,59, and 57 mosquitoes for the four populations, for which we detected L1014F mutation frequencies of 14.29%, 35.00%, 20.34%, and 31.58%, respectively, with a majority of these mutations being heterozygous for resistance. In addition, five adult mosquitoes were identified has having synonymous mutations at site 1011[i. e. , AAT(asparagine)mutation to AAC(asparagine)]. Conclusions Our findings revealed the clear resistance of Cx. tritaeniorhynchus to deltamethrin in the Yancheng region of the Yellow Sea wetland, and the resistance phenotype and kdr frequency of Cx. tritaeniorhynchus in the wetland environment were comparable to those of Cx. tritaeniorhynchus in the wetland environment, thereby indicating that the resistance of different populations of Cx. tritaeniorhynchus was homogeneous under the pressure of different insecticide selection within and around the wetland. However, the underlying mechanisms need to be further studied.
2."Compatibility" Relationship of Active Components and Heat-clearing and Blood-cooling Effect of Rehmannia glutinosa Roots
Yaman CHEN ; Jinpeng CUI ; Juan ZHANG ; Qingpu LIU ; Haiyan GONG ; Jingwei LEI ; Fengqing WANG ; Caixia XIE
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(12):193-201
ObjectiveTo analyze the "compatibility" relationship of sugars and glycosides and the heat-clearing and blood-cooling effect of the roots of four varieties of Rehmannia glutinosa and provide a basis for research on the pharmacodynamic material basis and quality control of R. glutinosa. MethodsThe content of sugars and glycosides in the roots of four varieties of R. glutinosa was determined during the growth period. The principal component analysis (PCA), orthogonal partial least squares-discriminant analysis (OPLS-DA), and the "compatibility" relationship of active components were employed to screen out the differential samples. A rat model of bleeding due to blood heat was used to verify the pharmacodynamic differences and the potential active components of differential samples. ResultsThe content and proportion characteristics of various components in roots of the four varieties of R. glutinosa during the expansion stage and the maturity stage had obvious differences. The proportion of phenylethanoid glycosides at the maturity stage was higher than that at the expansion stage. The R. glutinosa variety 85-5 had special quality characteristics among the tested varieties. All the samples alleviated the symptoms in the rat model. The effect of clearing heat and cooling blood was different between the maturity stage and the expansion stage, as well as between 85-5 samples at the maturity stage and other samples. The effect of clearing heat and cooling blood of R. glutinosa roots was the result of the combined action of multiple components in R. glutinosa roots and might be related to the high proportions of polysaccharides, iridoid glycosides, and phenylethanoid glycosides. ConclusionThe growth stage and variety affect the quality of R. glutinosa roots. The effect of clearing heat and cooling blood of R. glutinosa roots was related to the content and proportions of various components. The study can provide a basis for the basic research on the active components and quality control of R. glutinosa.
3."Compatibility" Relationship of Active Components and Heat-clearing and Blood-cooling Effect of Rehmannia glutinosa Roots
Yaman CHEN ; Jinpeng CUI ; Juan ZHANG ; Qingpu LIU ; Haiyan GONG ; Jingwei LEI ; Fengqing WANG ; Caixia XIE
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(12):193-201
ObjectiveTo analyze the "compatibility" relationship of sugars and glycosides and the heat-clearing and blood-cooling effect of the roots of four varieties of Rehmannia glutinosa and provide a basis for research on the pharmacodynamic material basis and quality control of R. glutinosa. MethodsThe content of sugars and glycosides in the roots of four varieties of R. glutinosa was determined during the growth period. The principal component analysis (PCA), orthogonal partial least squares-discriminant analysis (OPLS-DA), and the "compatibility" relationship of active components were employed to screen out the differential samples. A rat model of bleeding due to blood heat was used to verify the pharmacodynamic differences and the potential active components of differential samples. ResultsThe content and proportion characteristics of various components in roots of the four varieties of R. glutinosa during the expansion stage and the maturity stage had obvious differences. The proportion of phenylethanoid glycosides at the maturity stage was higher than that at the expansion stage. The R. glutinosa variety 85-5 had special quality characteristics among the tested varieties. All the samples alleviated the symptoms in the rat model. The effect of clearing heat and cooling blood was different between the maturity stage and the expansion stage, as well as between 85-5 samples at the maturity stage and other samples. The effect of clearing heat and cooling blood of R. glutinosa roots was the result of the combined action of multiple components in R. glutinosa roots and might be related to the high proportions of polysaccharides, iridoid glycosides, and phenylethanoid glycosides. ConclusionThe growth stage and variety affect the quality of R. glutinosa roots. The effect of clearing heat and cooling blood of R. glutinosa roots was related to the content and proportions of various components. The study can provide a basis for the basic research on the active components and quality control of R. glutinosa.
4.The Relationship Between Atherogenic Index of Plasma and Rapid Progression of Coronary Non-target Lesions
Wei WANG ; Haobo XU ; Juan WANG ; Jiansong YUAN ; Weixian YANG ; Rong LIU ; Shubin QIAO ; Jingang CUI
Chinese Circulation Journal 2025;40(11):1076-1080
Objectives:The study assessed the relationship between atherogenic index of plasma(AIP)and the rapid progression of coronary non-target lesions.Methods:A total of 1 247 patients with coronary artery disease who underwent two coronary angiography examinations at Fuwai Hospital,Chinese Academy of Medical Sciences between January 2010 and September 2014 were enrolled in this retrospective study.The AIP is defined as the base 10 logarithm of the ratio of the concentrations of triglyceride to high-density lipoprotein cholesterol.Patients were divided into the high AIP group(n=623)and the low AIP group(n=624)based on the median value of AIP.Lesion rapid progression is defined as an increase of more than 10%in the lumen stenosis of the lesion with a stenosis rate of more than 50%,or an increase of more than 30%in the lumen stenosis rate of the lesion with a stenosis rate of less than 50%,or a progression to total occlusion within 2 years.Results:Median AIP was 0.39(0.23-0.56)in this patient cohort.Rapid progression of non-target lesions occurred in 65(5.21%),including 42(6.74%)in the high AIP group.The Kaplan-Meier curve showed that the cumulative incidence of rapid progression of non-target lesions was higher in the high AIP group than in the low AIP group(HR=1.751,95%CI:1.053-2.912,log-rank P=0.028).In univariate cox analysis,the AIP and high AIP correlated with rapid progression of non-target lesions.After multivariate adjustment,AIP was an independent risk factor for rapid progression of non-target lesions(adjusted HR=2.731,95%CI:1.090-6.844,P=0.032).Conclusions:AIP is an independent risk factor for rapid progression of non-target lesions.AIP should be considered as a biomarker for estimating the risk of cardiovascular disease,along with other traditional risk factors.
5.Effects of irbesartan regulating SDF-1/CXCR4 pathway on proliferation,migration and radiosensitivity of lung cancer cells
Dong-juan WANG ; Xiang-yao LIAN ; Cui-min ZHU ; Xi-ying LYU ; Ping-ping LIN
Journal of Regional Anatomy and Operative Surgery 2025;34(3):192-198
Objective To analyze the effects of irbesartan(IBN)regulating the stromal cell-derived factor-1(SDF-1)/CXC chemokine receptor 4(CXCR4)pathway on proliferation,migration,and radiosensitivity of lung cancer cells.Methods Human lung cancer A549 cells were randomly divided into the A549 group(without treatment),radiation group(4 Gy X-ray radiation),IBN group(1 μmol/L IBN treatment for 24 hours),IBN+radiation group(1 μmol/L IBN treatment for 24 hours+4 Gy X-ray radiation),pcDNA3.1 group(transfected with pcDNA3.1+1 μmol/L IBN treatment for 24 hours+4 Gy X-ray radiation),and SDF-1 group(transfected with pcDNA3.1 SDF-1+1 μmol/L IBN treatment for 24 hours+4 Gy X-ray radiation).The cell viability,colony formation,apoptosis,and migration of each group were observed.The leakage rate of lactate dehydrogenase(LDH)and Ang Ⅱ levels in cells were detected.Immunofluorescence method was applied to analyze the number of γH2AX focal points of cells in each group.Western blot was applied to detect the expression of proliferation and apoptosis related proteins and SDF-1/CXCR4 pathway related proteins.Results Both radiation and IBN treatment inhibited the proliferation and migration of A549 cells,promoted cell apoptosis,upregulated the number of γH2AX focal points and LDH leakage rate,upregulated the expression of Caspase-3,Bax,and Caspase-7,and downregulated the level of Ang Ⅱand expression of SDF-1,CXCR4,Bcl-2 and PCNA(P<0.05).The combined treatment of radiation and IBN further enhanced the changes of the above indicators(P<0.05).And SDF-1 treat-ment effectively reversed the effects of radiation and IBN treatment on the changes of the above indicators(P<0.05).Conclusion IBN can limit proliferation and migration of lung cancer cells,and increase radiosensitivity by inhibiting the SDF-1/CXCR4 pathway.
6.Changing trends of isolation rates of multidrug-resistant organisms in Shaanxi Province from 2014 to 2023 and comparison with national average levels
Qiaodi GUI ; Yanyan GONG ; Cui WANG ; Juan MA
Chinese Journal of Nosocomiology 2025;35(21):3304-3309
OBJECTIVE To investigate the changes of isolation rates of multidrug-resistant organisms(MDROs)in Shaanxi Province from 2014 to 2023 so as to provide theoretical bases for prevention,control and surveillance of drug-resistant organisms.METHODS The data of MDROs that were detected by the networked units of Shaanxi Antimicrobial Resistance Surveillance System in 10 years from 2014 to 2023 were analyzed by WHONET 5.6 soft-ware and were compared with the national average levels.RESULTS The isolation rates of MDROs generally showed downward trends from 2014 to 2023(all P<0.05).The isolation rate of methicillin-resistant Staphylo-coccus aureus(MRSA)dropped from 45.7%to 35.8%,and that of methicillin-resistant coagulase-negative Staph-ylococcus(MRCNS)decreased from 81.8%to 75.8%,higher than the national average levels.The isolation rate of vancomycin-resistant Enterococcus faecium(VREfm)dropped from 3.1%to 0.5%,and that of vancomycin-re-sistant Enterococcus faecalis(VRE fa)declined from 1.7%to 0,equal to or slightly lower than the national aver-age levels.Among the Streptococcus pneumoniae strains,the isolation rate of erythromycin-resistant S.pneumon-iae(ERSP)was higher than that of the penicillin-resistant S.pneumoniae(PRSP),the isolation rate of PRSP showed a downward trend(from 8.0%to 0.8%),while the isolation rate of ERSP rose slightly(from 96.2%to 98%),slightly higher than the national average levels.The isolation rate of carbapenem-resistant Escherichia coli(CR-ECO)showed a downward trend(from 2.3%to 1.0%),the isolation rate of carbapenem-resistant Klebsiella pneumoniae(CR-KPN)fluctuated and showed slightly rising trend(from 6.1%to 8.2%),slightly lower than the national average levels.The isolation rates of carbapenem-resistant Acinetobacter baumannii(CR-ABA)and carbapenem-resistant Pseudomonas aeruginosa(CR-PAE)strains showed downward trends(with CR-ABA de-creasing from 66.2%to 60.3%,CR-PAE declining from 32.6%to 19.7%),slightly higher than the national aver-age levels.CONCLUSIONS The isolation rates of the MDROs showed downward trends in Shaanxi Province from 2014 to 2023 except for ERSP and CRKP.The isolation rates of VRE and CRE are lower than the national average levels,while the isolation rates of other MDROs are slightly higher than the national average levels.
7.Pathological diagnosis of solid pancreatic lesions with endoscopic ultrasound-guided fine needle aspiration: a series study of 311 cases
Xiaoxiao WEN ; Xiaoyi LIU ; Jinfeng CUI ; Lichao ZHANG ; Wenxuan LIU ; Haiyan YANG ; Yuan WANG ; Li YI ; Lei LOU ; Juan WANG ; Yuehong LI ; Wenxin WU ; Xianghong ZHANG
Chinese Journal of Pathology 2025;54(1):52-58
Objective:To investigate the combined application of cytology, cell block histology and immunohistochemistry to improve the diagnostic accuracy of solid pancreatic lesions in endoscopic ultrasound-guided fine needle aspiration (EUS-FNA) samples.Methods:The pathological data of EUS-FNA in 311 cases of solid pancreatic lesions submitted to the Second Hospital of Hebei Medical University, Shijiazhuang, China from May 2019 to September 2023 were retrospectively analyzed. The cases included pancreatic ductal adenocarcinoma (PDAC, 172 cases), solid pseudopapillary neoplasm (SPN, 12 cases), neuroendocrine tumors (PNET, 14 cases) and chronic pancreatitis (113 cases). The cytological features of smears, the histology of cell block sections and the diagnostic markers in PDAC, SPN and PNET were analyzed. The diagnostic accuracies of cytology, cell block histology/immunohistochemistry and combination of the two methods for classifying these pancreatic solid lesions were evaluated.Results:Irregular arrangement of atypical (cancer) cells, anisonucleosis and nuclear atypia were the typical cytological features of PDAC, while presence of pseudopapillae with a myxoid/hyalinized fibrovascular core and low adhesion/salt-and-pepper chromatin were diagnostic features of SPN and NET, respectively. Immunohistochemical results showed that CK7 and CK19 were the most sensitive markers of pancreatic ductal epithelia, and the diffuse strong expression of S-100P (102/111, 91.9%) and aberrant expression of p53 (80/111, 72.1%) were important immunophenotypic markers of PDAC. Various degrees of CDX2 expression could be found in 66.4% PDAC. The expression of CD10, PR, vimentin, CD99 and cyclinD1 and the aberrant expression of β-catenin were the immunophenotypic features of SPN, while the expression of CgA, Syn and CD56 were indispensable immunemarkers for the diagnosis of PNET. Overall, cytology had higher sensitivity than cell block histology (93.9% versus 82.8%) and lower specificity (92.9% versus 99.1%), while the combination of the two methods significantly improved the sensitivity to 96.9% in solid pancreatic lesions. The combination of cytology and cell block histology could significantly improve the diagnostic efficacy of EUS-FNA in PDAC.Conclusions:Integrated diagnosis based on cytology (including rapid on-site evaluation), cell block histology and immunohistochemical findings could significantly improve the diagnostic yield of EUS-FNA in classifying solid pancreatic lesions.
8.The efficacy of plasma gasdermin D C-terminal fragment in the early diagnosis of sepsis
Yuexian LYU ; Xiu BI ; Ying LIU ; Shujing CUI ; Lixin ZHAO ; Ge GAO ; Jianxia WANG ; Juan LI ; Jun LI
The Journal of Practical Medicine 2025;41(12):1899-1906
Objective To assess the effectiveness of the Gasdermin D C-terminal fragment(GSDMD-CT)as a novel plasma biomarker for the clinical diagnosis of sepsis.Methods Between July 2021 and November 2024,245 patients from Tangshan Gongren Hospital were enrolled in this study.In accordance with the diagnostic criteria for sepsis and the systemic inflammatory response syndrome(SIRS),patient samples were classified into the sepsis group and the SIRS group.Meanwhile,healthy individuals were selected as the healthy control(HC)group.Sepsis patients were further categorized into the Gram-positive bacterial group,the Gram-negative bacterial group,and the fungal group based on the type of pathogen infection.The levels of GSDMD-CT,C-reactive protein(CRP),and procalcitonin(PCT)were measured in all subjects.Nonparametric tests were employed to compare the differences in various indices among different groups.The diagnostic value of GSDMD-CT in sepsis was evalu-ated by constructing the receiver operating characteristic(ROC)curve.Spearman's correlation analysis was used to examine the relationships among GSDMD-CT,CRP,and PCT.Results The plasma GSDMD-CT levels in the sepsis group 23.02(16.71,33.01)pg/mL and in the SIRS group 16.52(11.26,22.22)pg/mL were significantly higher than those in the healthy control group 7.02(4.42,11.43)pg/mL(U=-10.175,-7.890,P<0.001).Moreover,the plasma GSDMD-CT levels in the sepsis group were significantly higher than those in the SIRS group(U=-2.941,P<0.05).In the Gram-positive bacterial group,the Gram-negative bacterial group,and the fungal group,the GSDMD-CT levels were 23.01(17.16,27.51)pg/mL,23.41(16.78,35.50)pg/mL,and 16.29(14.53,56.27)pg/mL,respectively.When compared with the healthy control group,the GSDMD-CT levels in these three groups were all significantly higher(P<0.05).However,there were no significant differences in GSDMD-CT levels among these three groups(P>0.05).The area under the curve(AUC)of plasma GSDMD-CT for diagnosing sepsis was 0.881(95%confidence interval:0.833~0.929),with a Youden index(YI)of 0.695,a sensitivity of 85.0%,and a specificity of 84.5%.Spearman correlation analysis indicated a weak correlation between GSDMD-CT and C-reactive protein(CRP)(r=0.32,P<0.001)and a positive correlation between GSDMD-CT and procalci-tonin(PCT)(r=0.65,P<0.001).Conclusion GSDMD-CT exhibits significant clinical value in the diagnosis of sepsis and holds great potential as a biomarker in the diagnostic process of sepsis.
9.Evaluation of left ventricular function changes in pregnant women with different severities of hypertensive disorders of pregnancy by strain and tissue-tracking mitral annular displacement parameters
Ziyue WANG ; Xinxia WANG ; Hezhou LI ; Juan WU ; Junling ZHANG ; Yun LIU ; Yu CUI ; Manyu LI ; Yulu MIAO
Chinese Journal of Ultrasonography 2025;34(9):784-791
Objective:To evaluate the changes in left ventricular structure and function in pregnant women with different severities of hypertensive disorders of pregnancy(HDP)using conventional echocardiographic parameters,strain,and tissue-tracking mitral annulus displacement(TMAD)parameters,and to validate and compare the application value of strain and TMAD parameters.Methods:A total of 148 singleton pregnant women with HDP and 100 healthy pregnant women(HP group)who attended the Third Affiliated Hospital of Zhengzhou University from October 2023 to July 2024 were selected. Conventional echocardiographic parameters,strain,and TMAD parameters were collected. Based on the severity of the disease,HDP patients were divided into the gestational hypertension group(GH group, n=49),non-severe preeclampsia group(NSPE group, n=35),and severe preeclampsia group(SPE group, n=64). The differences in various parameters between the HP,GH,NSPE,and SPE groups were compared. The correlation between the displacement ratio of the midpoint of the mitral valve annulus in the apical 4-chamber(AP4 Midpt%),the displacement ratio of the midpoint of the mitral valve annulus in the apical 2-chamber(AP2 Midpt%)and left ventricular global longitudinal strain(LVGLS)were analyzed,and ROC curves were plotted to analyze and compare the diagnostic efficacies of LVGLS,AP4 Midpt% and AP2 Midpt% for left ventricular function changes in HDP pregnant women. Results:① The analysis revealed no statistically significant differences in maternal age and height between the HP group and the HDP subgroups(all P>0.05). In contrast,statistically significant differences were observed in gestational age,systolic blood pressure,diastolic blood pressure,body mass index,and body surface area(all P<0.05). Additionally,significant differences were noted in left ventricular mass,left ventricular interventricular septum thickness at end-diastole,left ventricular posterior wall thickness at end-diastole,left ventricular end-diastolic dimension,left ventricular end-systolic dimension,left ventricular end-diastolic volume,left ventricular end-systolic volume,and stroke volume between the HP group and the HDP subgroups(all P<0.05),while the difference in left ventricular ejection fraction was not statistically significant( P>0.05). ② Significant differences were identified in strain and TMAD parameters between the HP group and the HDP subgroups(all P<0.05),with LVGLS,AP4 Midpt% and AP2 Midpt% exhibited the largest effect sizes( η p2=0.457,0.453,0.351). A progressive decline in strain and TMAD parameters was observed as the severity of HDP increased. ③ There were strong positive correlations between AP4 Midpt%,AP2 Midpt% and LVGLS( r=0.752,0.747;all P<0.001). ④ LVGLS,AP4 Midpt% and AP2 Midpt% all demonstrated significant diagnostic efficacies for changes in left ventricular function in HDP(AUC=0.840,0.847,0.791),and the differences in AUC among the 3 curves were not statistically significant(all P>0.05).⑤ The success rate of collecting TMAD parameters was significantly higher than that of strain parameters(99.24% vs. 93.58%, P<0.001). Conclusions:As the severity of the disease worsens,the changes of left ventricular structure and function in pregnant women with HDP become more and more significant. Both strain and TMAD parameters can early and sensitively identify the subclinical damage of left ventricular systolic function in HDP pregnant women,and both of them have the same diagnostic value. TMAD parameters can be used as a reliable substitute parameter of LVGLS in HDP pregnant women.
10.Three-dimensional ultrasound combined with microvascular flow imaging for prenatal diagnosis of fetal intracranial anomalies
Yuanjie CUI ; Cuixia GUO ; Zhen LI ; Juan ZHANG ; Tiejuan ZHANG ; Keyang WANG ; Qingqing WU ; Yuting WU ; Lijuan SUN
Chinese Journal of Medical Imaging Technology 2025;41(3):368-371
Objective To observe the value of three-dimensional ultrasound combined with microvascular flow imaging for prenatal diagnosis of fetal intracranial anomalies.Methods Totally 118 fetuses with intracranial anomalies diagnosed through cranial MRI/induced labor specimen dissection who underwent prenatal ultrasound examination were retrospectively enrolled.Two-dimensional,three-dimensional ultrasound and microvascular flow imaging manifestations of fetal intracranial anomalies were observed,and the accuracy of three-dimensional ultrasound combined with microvascular flow imaging for prenatal diagnosis of fetal intracranial anomalies was analyzed.Results The accuracy of three-dimensional ultrasound combined with microvascular flow imaging for prenatal diagnosis of fetal intracranial anomalies was 93.22%(110/118),of isolated and non-isolated intracranial anomalies was 97.47%(77/79)and 84.62%(33/39),respectively.Six fetuses were missed diagnosis of malformations of cortical development(1 fetus of tuberous sclerosis,4 of abnormal morphology of the sulci gyrus and 1 of schizencephaly)and 1 fetus of intracranial softening lesion,while posterior fossa arachnoid cyst in 1 fetus was misdiagnosed as mega cisterna magna.Conclusion The accuracy of three-dimensional ultrasound combined with microvascular flow imaging for prenatal diagnosis of fetal intracranial anomalies was relatively high.


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