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2021 Vol. 51, No. 1 Published: 2021-01-25   1 Correlation between 12 loci of single nucleotide polymorphisms and childhood acute lymphoblastic leukemia MO Yixiao, WANG Zhen, WANG Pan, SHEN Shuhong DOI: 10.3969/j.issn.2095-9400.2021.01.001 Objective: To identify genetic susceptibility genes and the loci of single nucleotide polymorphisms (SNPs) of acute lymphoblastic leukemia (ALL) in Chinese children, expecting to provide novel strategy for the diagnosis and treatment of ALL. Methods: Integrated fluidic circuits (IFC) was used, through the analysis of the JunoTM Fluidigm system, to examine 13 SNPs loci of the genetic susceptibility genes in 191 pediatric ALL patients and 191 controls. After Hardy-Weinberg equilibrium test, the data was analyzed under case-control study using χ2 test and logistic regression. Results: The rs10849033 locus was eliminated because of inconformity with Hardy-Weinberg genetic balance. There were significant differences between ALL patients and controls in the risk allele frequencies of gene ARID5B rs10821936 (C allele: 0.489, P<0.001, OR=1.692, 95%CI:1.265-2.264), rs10994982 (A allele: 0.598, P=0.016, OR=1.425, 95%CI: 1.068-1.901); gene ZNF230 rs2191566 (A allele: 0.721, P=0.002, OR=1.599, 95%CI=1.179-2.169); gene IKZF1 rs11978267 (G allele: 0.191, P=0.001, OR=1.965, 95%CI=1.301-2.968); rs4132601 (G allele: 0.175, P=0.003, OR=1.871, 95%CI=1.225-2.857). Conclusion: We identified rs2191566 of gene ZNF230 and rs10821936, rs10994982 of gene ARID5B as the susceptibility loci of ALL in Chinese children. Rs4132601 and rs11978267 of gene IKZF1 were identified as the potential ALL susceptibility loci in Chinese children. 2021 Vol. 51 (1): 1-7 [Abstract] ( 925 ) HTML (1 KB)  PDF (1336 KB)  ( 833 ) 8 Association between insulin-like growth factor-2 in cord vein and macrosomia WANG Dan, YAN Ya’na,FAN Shuli, CAI Caiping, WANG Qiu DOI: 10.3969/j.issn.2095-9400.2021.01.002 Objective: To evaluate the serum concentration of insulin-like growth factor-2 (IGF-2) in cord vein and to assess its relationship with macrosomia. Methods: Totally 61 macrosomia (MAC) and 60 normal birth weight (NBW) term infants, born in the First Affiliated Hospital of Wenzhou Medical University from January 2018 to December 2019 were enrolled in this study. The serum IGF-2 concentration in cord vein was detected. Pearson or Spearman correlation analysis was used to assess the relationship between IGF-2 and growth parameters of the newborn. Results: The serum concentrations of IGF-2 in cord vein were (221.92±29.97)ng/mL and (169.69±14.53)ng/mL in MAC group and NBW group respectively, with significant difference (P<0.001). Newborns of gestational diabetes mellitus (GDM) versus euglycemic mothers had an elevated level of cord vein IGF-2 (GDM-MAC vs. nGDM-MAC=229.71 ng/mL vs. 214.18 ng/mL, P=0.046; GDM-NBW vs. nGDM-NBW=175.69 ng/mL vs. 161.48 ng/mL, P<0.001). Correlation analysis showed that cord vein IGF-2 was positively related to birth weight, birth length, and ponderal index (PI) (r=0.678, 0.566, 0.600, P<0.001). The concentration of IGF-2 was also related to gestational age (r=0.273, P=0.002). However, there was no significant correlation between IGF-2 level and newborn gender, maternal age, maternal higher education or primiparity (P>0.05). Conclusion: IGF-2 level in cord vein of newborn is affected by maternal gestational diabetes mellitus and gestational age. Meanwhile, IGF-2 is correlated to fetal growth and its high level may play an important role in the occurrence of macrosomia. 2021 Vol. 51 (1): 8-12，18 [Abstract] ( 665 ) HTML (1 KB)  PDF (1586 KB)  ( 731 ) 13 Cloning of the cDNA fragment coding for rat ENPP1 protein and construction of recombinant adeno-associated virus DI Feng, WU Xiujuan, SHEN Shuijuan, GUO Boyao DOI: 10.3969/j.issn.2095-9400.2021.01.003 Objective: To clone the full-length cDNA fragment coding of rat ENPP1 and construct the recombinant adeno-associated virus vector of ENPP1. Methods: Using rat vascular smooth muscle cell mRNA as templates, the large fragment of rat ENPP1 gene was obtained through RT-PCR amplification and cloned into pCDH vector. The fragment was then cloned into adeno-associated virus vector (AVV) vector pAAV-MCS by EcoR I/Xho I enzyme digestion, ligation and transformation, after which the positive clone was identified by enzyme digestion and DNA sequencing. Recombinant AAV-ENPP1 was packaged and introduced to the targeted vascular smooth muscle cells. Western blot was used to detect the expression of target ENPP1 protein. Results: The 2 721 bp large fragment of rat ENPP1 gene was successfully cloned and constructed using RT-PCR technique, and finally obtained pAAV/ENPP1 clone with correct sequencing. The results of Western blot testing revealed that the expression of the target protein ENPP1 was increased. Conclusion: The successful construction of recombinant adeno-associated virus vector of rat ENPP1 lays a foundation for the treatment of vascular calcification via ENPP1 gene transfection. 2021 Vol. 51 (1): 13-18 [Abstract] ( 714 ) HTML (1 KB)  PDF (2191 KB)  ( 1067 ) 19 Clinical value of SNP array in prenatal diagnosis of fetus with increased nuchal translucency ZHOU Lili, WEN Zhengjing, XU Chenyang, LI Huanzheng, XU Xueqin, TANG Shaohua DOI: 10.3969/j.issn.2095-9400.2021.01.004 Objective: To investigate the clinical value of SNP array analysis in prenatal diagnosis of fetuses with increased nuchal translucency. Methods: A total of 220 fetus with increased NT (NT≥2.5 mm) were tested by SNP array and conventional karyotyping. Results: Chromosomal abnormality was identified in 14.5% (32/220) fetus, with aneuploidy accounting for 87.5% and structural alterations accounting for 12.5%. Among them trisomy 21 was the most common (43.8%), followed by trisomy 18 (21.9%). Infetuses with normal karyotype, SNP array detected another nine cases with abnormal CNVs, of which five cases were pathogenic CNVs and four cases were likely pathogenic CNVs. The fetuses were classified into three groups according to NT thickness 2.5-2.9 mm, 3.0-3.4 mm and ≥3.5 mm, with the abnormal karyotype rate being 5.6%, 7.9% and 23.3% respectively and the abnormal SNP array rate being 5.6%, 11.1% and 30.1% respectively. The chromosome abnormality rate increased significantly with the increase of NT thickness. The application of SNP array analysis for fetuses with increased NT (≥3.5 mm) could detect another seven abnormal CNVs, improving the genetic etiology rate by 6.8%, which had statistical difference compared with chromosomal karyotype (P<0.05). However, there was no statistical difference when NT thickness was 3.0-3.4 mm. Conclusion: NT thickness is correlated with chromosome abnormality in that fetal chromosomal abnormal rate increases significantly with the increase of NT thickness. The application of SNP array analysis for fetuses with increased NT (≥3.5 mm) in prenatal diagnosis could improve the genetic etiology rate by 6.8%, which is of great value to clinical genetic counseling.   2021 Vol. 51 (1): 19-24 [Abstract] ( 687 ) HTML (1 KB)  PDF (1575 KB)  ( 848 ) 25 Epidemiological characteristics of human adenovirus in hospitalized children with lower respiratory tract infection and its correlation with meteorological factors in Wenzhou WEN Shunhang, LIN Zupan, ZHOU Bigao, CHEN Xiaofang, ZHENG Jingwei, ZHANG Hailin. DOI: 10.3969/j.issn.2095-9400.2021.01.005 Objective: To study the epidemiological characteristics of human adenovirus (HAdV) infection in lower respiratory tract among hospitalized children in Wenzhou and its correlation with meteorological factors. Methods: A retrospective analysis was made on respiratory virus antigen detection data of children admitted to the Second Affiliated Hospital & Yuying Children’s Hospital of Wenzhou Medical University between Jan 2008 and Dec 2017 due to acute lower respiratory tract infection, and the epidemiological characteristics of HAdV infection were analyzed. Meanwhile, meteorological data of Wenzhou from Jan 2008 to Dec 2017 were collected, including average temperature per ten-day period, average humidity, total rainfall, rainfall days and average wind speed. Generalized additive models (GAM) were used to conduct univariate and multivariate regression analyses to explore the correlation between HAdV prevalence and meteorological factors. Results: Of 89 898 nasopharyngeal aspirates and/or sputum specimens tested for respiratory virus antigen between Jan 2008 and Dec 2017, a total of 1 974 (2.2%) cases tested positive for HAdV infection, of which 339 cases were <1 year old, 615 cases were 1-3 years old, 597 cases were 3-5 years old, and 423 cases were aged 5-18 years. The number and rate of HAdV detection in children peaked between May and July. Based on GAM moder, HAdV detection rate increased with the temperature getting higher (P=0.015) and the wind speed getting slower (P=0.003). With the increase of wind speed, the decline scale of detection rate reduced gradually. Conclusion: HAdV infection peaked in late spring and early summer among hospitalized children with lower respiratory tract infection in Wenzhou. Temperature and wind speed are the main meteorological factors related to the prevalence of HAdV. 2021 Vol. 51 (1): 25-29 [Abstract] ( 729 ) HTML (1 KB)  PDF (1688 KB)  ( 1031 ) 30 Diagnostic value of the third generation Force dual energy CT in contusion and occult fracture of sacral pterygoid bone XIA Haihong, GAO Yang, XIAO Yangrui, LIU Weiwen, YE Yongjun, CHEN Tao, ZHANG Wenwei, JI Jiansong. DOI: 10.3969/j.issn.2095-9400.2021.01.006 Objective: To explore the value of the third generation Force dual energy CT virtual non-calcium (DECT-VNCa) map in the diagnosis of contusion and occult fracture of sacral alar under the condition of different relative contrast material ratio (RCMR). Methods: Totally 35 patients with definite history of pelvic trauma or lumbosacral pain in the past 3 years were prospectively collected for dual source CT dual energy scan and MRI examination. The RCMR values were 1.59, 1.79, 1.89, respectively. Three groups of DE-VNCa images were reconstructed. Taking MRI diagnosis as the gold standard, the efficacy of three groups of images was calculated in the diagnosis of sacral pterygoid bone contusion and occult fracture, and the optimal RCMR value was obtained. Under this optimal condition, the CT values of sacral alar bone marrow edema area and contralateral normal area were measured and compared, and then the diagnostic efficacy of CT value in sacral alar occult fracture was evaluated by ROC curve. Results: When the RCMR values was 1.59, 1.79 and 1.89, the diagnostic efficacy of DE-VNCa was the highest, when Yoden index for the diagnosis of sacral alar bone marrow edema was 0.40, 0.81 and 0.71 respectively. Using RCMR=1.79 to reconstruct DE-VNCa map, the average CT value of bone marrow edema area was (-20.21±18.81) Hu, and the average CT value of contralateral normal bone marrow area was (-74.19±27.94) Hu. There was significant difference between sacral alar bone marrow edema area and contralateral normal area (P<0.001). Taking CT=-43.45 Hu as the critical value, the AUC for diagnosis of sacral alar bone marrow edema was 0.947 (P<0.001). Conclusion: Dual source CT DE-VNCa is of high clinical value in the diagnosis of sacral pterygoid bone contusion and occult fracture when RCMR is 1.79, and CT value is helpful in the diagnosis of sacral pterygoid bone contusion and bone marrow edema of occult fracture. 2021 Vol. 51 (1): 30-34 [Abstract] ( 829 ) HTML (1 KB)  PDF (2270 KB)  ( 860 ) 35 Expression of SOAT1 and its clinical significance in hepatocellular carcinoma CHEN Yao, YANG Xunjun, CHEN Hanbin, CHEN Guorong DOI: 10.3969/j.issn.2095-9400.2021.01.007 Objective: To investigate the expression of sterol O-acyltransferase 1 (SOAT1) in hepatocellular carcinoma (HCC) tissues and tissues adjacent to the cancer, and to explore its correlation with patient’s prognosis and clinicopathological features. Methods: The HCC dataset was collected from the Cancer Genome Atlas (TCGA) database, and download the expression of SOAT1 mRNA and prognostic data. The expression of SOAT1 mRNA in 50 normal liver tissues and 374 HCC tissues was compared to analyze the correlation between expression and prognosis. Collected paraffin tissue specimens of 53 HCC patients from the First Affiliated Hospital of Wenzhou Medical University from May 2018 to October 2019, using immunohistochemical (IHC) methods to detect the protein expression level of SOAT1 in cancer and adjacent tissues, and its correlation with clinical pathology features was analyzed. Results: Data analysis of TCGA database showed that compared with normal tissues adjacent to cancer, SOAT1 expression was significantly increased in HCC (P<0.001). The prognostic survival analysis found that 5-year survival rate of high expression of SOAT1 was lower (P<0.05). IHC results showed that compared with adjacent tissues, the expression of SOAT1 protein in HCC tissues was significantly increased (P<0.05). The expression of SOAT1 protein in cancer tissues was correlated with the clinical stage of the tumor and microvascular tumor thrombosis (P<0.05). Conclusion: The expression of SOAT1 is increased in HCC tissues, and closely related to the clinicopathological characteristics of HCC cancer patients, with its expression highly correlated with the malignancy degree of the tumor. 2021 Vol. 51 (1): 35-39 [Abstract] ( 815 ) HTML (1 KB)  PDF (1484 KB)  ( 1298 ) 40 The mechanism of epithelial-mesenchymal transition of human intrahepatic biliary epithelial cells induced by hypoxia/hypoxic inducible factor-1α through tgf-β1 YANG Zaixing, LIANG Yan, LIU Donghong, ZHANG Zhencheng, LUO Wanwan, MAO Panying. DOI: 10.3969/j.issn.2095-9400.2021.01.008 Objective: To investigate the role of hypoxia/hypoxic inducible factor-1α (HIF-1α) in inducing epithelial-mesenchymal transition of human intrahepatic biliary epithelial cells (HIBEC) as well as its molecular mechanisms. Methods: Under the condition of hypoxia (1% O2), HIBECs were cultured in vitro and analyzed for mobility, invasiveness and expression levels of EMT-related markers including E-cadherin and S100A4. The above-mentioned experiments were repeated with HIF-1α silenced by siRNA and transforming growth factor β1 (TGF-β1) inhibited by inhibitor LY364947. Results: Under the condition of hypoxia, in contrast to normoxia, HIBECs showed markedly enhanced mobility and invasiveness as well as significant downregulation of E-cadherin and upregulation of S100A4. After HIF-1α silence or TGF-β1 inhibition, above-mentioned biological changes of HIBECs were significantly inhibited under hypoxia. Conclusion: Hypoxia can induce EMT of HIBECs by activating HIF-1α. TGF-β1 plays an important role in hypoxia/HIF-1α-induced EMT of HIBECs.   2021 Vol. 51 (1): 40-44 [Abstract] ( 727 ) HTML (1 KB)  PDF (3499 KB)  ( 742 ) 45 The expression of EGFL6 in gastric cancer and its relationship with prognosis LI Liyi, TU Fuyang, YOU Sunwu, SHEN Leibin, YOU Tao, ZHENG Zhiqiang. DOI: 10.3969/j.issn.2095-9400.2021.01.009 Objective: To explore the expression of epidermal growth factor-like (EGFL) 6 in gastric cancer and its relationship with clinicopathological factors and prognosis. Methods: Eighty cases of primary gastric cancer specimens and adjacent tissues were collected from the Second Affiliated Hospital & Yuying Children’s Hospital of Wenzhou Medical University from March 2016 to February 2018. The expression of EFGL6 in tissues was determined by immunohistochemical staining. Statistical analysis was performed on the staining results and related clinical pathological factors. Results: The expression of EGFL6 was correlated with tumor diameter, depth of invasion, lymph node metastasis and TNM stage (P<0.05). The expression of EGFL6, patient age, tumor diameter, cell differentiation, depth of invasion, lymph node metastasis, and TNM stage were associated with the prognosis of patients with gastric cancer (P<0.05). Conclusion: The expression of EGFL6 is correlated with clinicopathological factors of gastric cancer, suggesting that EGFL6 plays an important role in the occurrence and development of gastric cancer. 2021 Vol. 51 (1): 45-48 [Abstract] ( 697 ) HTML (1 KB)  PDF (1423 KB)  ( 1012 ) 49 The effect of IGFBP-4 treatment on immunogenic cell death in A549 cells and its mechanism LAN Xiu, LI Weiwen, SUN Lei, LI Yuan, ZHAO Jialu. DOI: 10.3969/j.issn.2095-9400.2021.01.010 Objective: To investigate the effect of IGFBP-4 treatment on immunogenic cell death in A549 cells and its molecular mechanism. Methods: A549 cells were cultured in vitro and assigned to the control group, IGFBP-4 group, IGFBP-4+NAC group and IGFBP4+si-p38 group, respectively. After 48 hours, MTT was used to detect the survival rate of cells; flow cytometry was used to detect ROS content and calreticulin (CRT); ELISA was used to detect HMGB1 in medium and SOD and MDA in cells; Western blot was used to analyzed the expression of phosphorylated p38 (p-p38) and total p38. Results: Compared with control group, the content of ROS, CRT, HMGB1, MDA, the expression of p-p38 and p38 increased, while the survival rate and content of SOD2 decreased in IGFBP-4 group (P<0.05). Compared with IGFBP-4 group, the content of ROS, CRT, HMGB1, MDA decreased, while the survival rate and content of SOD2 increased in IGFBP4+NAC group and IGFBP4+si-p38 group (P<0.05). Conclusion: IGFBP-4 promoted immunogenic cell death via activating p38 MAPK signaling and inducing oxidative stress in A549 cells. 2021 Vol. 51 (1): 49-52,57 [Abstract] ( 873 ) HTML (1 KB)  PDF (1655 KB)  ( 1030 ) 53 MR enhancement features of small combined hepatocellular-cholangiocarcinoma and its pathological basis#br# XU Xiaofei, LI Bingrong, MAO Weibo, XIAO Yangrui, WANG Zufei. DOI: 10.3969/j.issn.2095-9400.2021.01.011 Objective: To investigate the MR enhancement features of small combined hepatocellular-cholangiocarcinoma (sCHC) and its pathological basis. Methods: The clinical and imaging data of 24 patients pathologically confirmed as sCHC from April 2011 to July 2019 in Lishui Central Hospital were retrospectively analyzed. The enhancement features of the lesions were mainly observed and compared to the pathological results. Then, Fisher test was used to analyze the different enhancement modes among the different dominant tumor cells types of lesion. Results: Six cases showed obvious thick-ring enhancement in the arterial phase and continuous enhancement in the following phase, whose pathology demonstrated as a circle of diffuse and mixed distribution of two types of survival tumor cells in the periphery of the lesion, with the center of the lesion mainly coagulative necrosis. One case showed obvious thick-ring enhancement in the arterial phase and filling enhancement in the following phase, whose pathology demonstrated as a circle of diffuse and mixed distribution of two types of living tumor cells in the periphery of the lesion, with the center of the lesion fibrous matrix and little coagulative necrosis. Fourteen cases showed significant entire enhancement in the arterial phase and continuous enhancement in the following phase, whose pathology demonstrated as diffuse and mixed distribution of two types of survival tumor cells in the lesion, in which fibrous matrix was not abundant, and with little or no coagulative necrotic necrosis. One case showed nodule-in-nodule enhancement, whose pathology demonstrated as migrating distribution of two types of survival tumor cells, with little fibrous matrix and no coagulative necrotic necrosis. Two cases showed reverse enhancement, whose pathology demonstrated as migrating distribution of two types of survival tumor cells, with more fibrous matrix in the ICC district than HCC district, and no coagulative necrosis in either of them. Of the 24 cases, 12 were ICC dominant, 8 were HCC dominant and 4 were proportional proximity. There was no significant difference in the enhancement performance of the proportion of sCHC (P>0.05). Conclusion: Different MR enhancement manifestations of sCHC lesions have their corresponding pathological basis. 2021 Vol. 51 (1): 53-57 [Abstract] ( 648 ) HTML (1 KB)  PDF (1504 KB)  ( 856 ) 58 The clinical value of elastography in the diagnosis of thyroid microcarcinoma CHEN Xiaoqin, SUN Haiqin, WENG Xijin, HE Jun, CHEN Limin. DOI: 10.3969/j.issn.2095-9400.2021.01.012 Objective: To explore the value of high frequency ultrasonography combined with elastography in the diagnosis of thyroid microcarcinoma. Methods: Totally 68 thyroid microcarcinoma patients were collected after fine needle biopsy or surgical treatment. High-frequency ultrasonic features, elastography score and strain rate ratio were retrospectively analyzed and qualitative diagnosis was made. The results were compared with those of surgery or fine needle puncture. Results: There were 87 nodules in 68 thyroid microcarcinoma patients of which 55 benign nodules and 32 thyroid cancers diagnosed by high-frequency ultrasound combined with elastography. Surgical or fine-needle aspiration results showed 59 benign nodules and 28 thyroid carcinomas. The sensitivity, specificity, accuracy, positive predictive value and negative predictive value of high-frequency ultrasound combined with elastography score in the diagnosis of minimal thyroid cancer were 89.2%, 89.8%, 89.7%, 80.6% and 89.9%, respectively, in contrast to 82.1%, 84.7%, 83.9%, 71.8% and 90.1% of thyroid microcarcinoma. Conclusion: High-frequency ultrasound is an important basis for thyroid nodule evaluation. The application of elastography can significantly improve the diagnostic value of ultrasound in the benign and malignant evaluation of thyroid small nodule. 2021 Vol. 51 (1): 58-61,65 [Abstract] ( 650 ) HTML (1 KB)  PDF (1390 KB)  ( 762 ) 62 An investigation into the complications of warfarin anticoagulant therapy and its influencing factors in patients with cardiac mechanical valve replacement SHI Huoshun, SHAO Guofeng, ZHAO Shunying, HU Xiaoyan. DOI: 10.3969/j.issn.2095-9400.2021.01.013 Objective: To investigate the influencing factors of warfarin-related complications in mechanical valve patients, so as to understand the factors that may influence patients' anticoagulation compliance. Methods: Patients who had mechanical valve replacement in Ningbo Medical Treatment Centre of Lihuili Hospital from January to December 2012 were enrolled in this investigation. The questionnaire, which included general information of patients, anticoagulant condition, coagulation function and other indicators, was completed by all subjects mainly via telephone visits and outpatient follow-up. Chi-square test was used for categorical variables,  and logistic regression model for multivariate analysis. Results: There was statistical difference in anticoagulant compliance and atrial fibrillation (AF) between two groups of anticoagulation complications (P<0.05), while there’s no significant difference between the two groups in the type of valve replacement, patient’s age, education, history of cerebral infarction, history of tobacco and alcohol and other long-term drug history (P>0.05). Multivariate logistic regression showed that AF and anticoagulation compliance were significantly associated with anticoagulant complications, with OR values being 2.570 and 4.628 respectively. Conclusion: Poor compliance with administration of warfarin and atrial fibrillation are independent risk factors for anticoagulation complications after mechanical valve replacement.   2021 Vol. 51 (1): 62-65 [Abstract] ( 805 ) HTML (1 KB)  PDF (1286 KB)  ( 956 ) 66 Effect of integrated vascular pathway team management on intravenous fistula of artificial vascular transplantation WU Yaoyao, JIN Lingwei, YE Bairu, CHEN Dongwei, GU He. DOI: 10.3969/j.issn.2095-9400.2021.01.014 Objective: To explore the effect of integrated management of vascular access team on artificial arteriovenous graft. Methods: From January 2017 to December 2018, 106 eligible hemodialysis patients who underwent continuous successful AVG were admitted to the Second Affiliated Hospital of Wenzhou Medical University. The participants were randomly divided into the control group and the study group by using a random number table. The control group was given routine vascular access management, and the study group was included in the management of the vascular access team. The participants were followed up prospectively to observe the fistula maturity rate and the primary patency rate. Kaplan-Meier method was used to estimate the AVG patency rate, and Log-Rank tests were used to compare survival curves. Results: The primary maturation rates of AVG were 77.8% (42/54) and 92.3% (48/52) in the control group and in the study group, respectively. There was a significant difference between the two groups (χ2=4.364, P=0.037). The one-year cumulative primary patency rates were 52.7% and 63.9% in the control group and in the study group, respectively. There was a significant differences in primary patency survival curves between the two groups (χ2=4.071, P=0.044). Conclusion: Establishing a vascular access team can improve the maturation rate and the patency rate of AVG. 2021 Vol. 51 (1): 66-69 [Abstract] ( 666 ) HTML (1 KB)  PDF (1333 KB)  ( 955 )

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