温州医科大学学报
 
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2022 Vol. 52, No. 1
Published: 2022-01-25
 
 
1 Baicalin inhibits hypoxia-induced pulmonary hypertension via the LncRNA-MALAT1/miR-361-3p/TGF-β pathway
WU Peiliang, CHEN Mayun, WANG Liangxing, HUANG Xiaoying
Objective: To explore the therapeutic effect of baicalin on hypoxic pulmonary hypertension and its potential mechanism. Methods: A total of eighteen male C57BL/6 mice were randomly divided into three groups: the normoxia group, hypoxia-saline group and hypoxia-baicalin group (n=6 in each group). After 21 days, right ventricular pressure (RVSP) and mean carotid artery pressure (mCAP) were detected by cardiac catheter. Hematoxylin-eosin (HE) staining was applied to visualize pulmonary artery remodeling and immunohistochemistry was used to detect the expression of TGF-β signaling pathway. Murine pulmonary artery smooth muscle cells (MPASMCs) were cultured for 48 h under hypoxic and normoxic conditions. CCK-8 assay was used to detect MPASMCs viability. The migration ability of MPASMCs was determined by wound healing assay and transwell assay. TGF-β pathway proteins expression was measured with Western blot analysis.Bioanalysis and dual luciferase reporter gene detection verify the regulatory relationship of LncRNA-MALAT1/miR-361-3p/TGF-β axis. Results: Compared with the hypoxia group, the baicalin group has significantly decreased proliferation and migration ability of MPASMCs (P<0.05), RVSP of mice was significantly decreased (P<0.05), and the pulmonary vascular remodeling was improved. Compared to the normoxic group, the hypoxiagroup had significantly increased expression of TGF-β signaling pathway related proteins TGF-β, p-smad2 and p-smad3, while the TGF-β signaling pathway was significantly inhibited after baicalin treatment. Mechanism exploration suggests that baicalin regulates the expression of TGF-β by regulating the LncRNA-MALAT1/miR-361-3p axis. Conclusion: Baicalin may regulate the expression of TGF-β by regulating the LncRNA-MALAT1/miR-361-3p axis, which can improve the abnormal proliferation and migration of hypoxic MPASMCs and attenuates pulmonary vascular remodeling in hypoxic pulmonary hypertension mice, thereby decreasing pulmonary arterial pressure.
2022 Vol. 52 (1): 1-8 [Abstract] ( 753 ) HTML (1 KB)  PDF (2251 KB)  ( 595 )
9 EPCs-Exo inhibit paclitaxel-induced pyrolysis of vascular endothelial cells
WANG Ping, MENG Liping,LIU Longbin, PENG Fang
DOI: 10.3969/j.issn.2095-9400.2022.01.002
Objective: To explore the effect of endothelial progenitor cell exosomes (EPCs-Exo) on paclitaxel-induced endothelial cell pyrolysis and its possible mechanism. Methods: The primary bone marrow mesenchymal stem cells (BMSCs) and ECPs were cultured and identified. The exosomes in the cultured medium of BMSCs and ECPs were extracted, and the morphology of the exosomes was observed by transmission electron microscope. The expression of CD63 and CD9 was detected by western blot to identify the exosomes. BMSCs-Exo and EPCs-Exo were used to intervene the endothelial cell pyrolysis model induced by paclitaxel, and the cell morphology was observed by microscope; Western blot was used to detect the exosomal marker proteins CD63,CD9 and pyrolysis-related proteins NLRP-3, IL-18, GSDMD-N. The ultrastructure change of the cells were observed under electron microscope. RNA sequencing was used to detect the difference of non-coding RNA in the two groups of exosomes, and qRT-PCR was performed to detect the expression of LncRNA FGD5-AS1 in each group. Results: BMSCs and EPCs were isolated, cultured and differentiated from rat bone marrow cells. The positiveness of CD90, CD44 in BMSCs and CD34, VEGFR2 in EPCs were confirmed by immunofluorescence.Compared with the control group, the cell membrane of the paclitaxel group was destroyed, the distance between mitochondria in the cell increased, the arrangement of intracellular fibers was disordered, and the expression of pyrooptosis proteins such as NLRP-3, IL-18, GSDMD-N was increased (P<0.05). Compared with the paclitaxel group, the cell membrane of the paclitaxel+EPCs-Exo group was more complete, the cell line damage was mild, and the expression of NLRP-3, IL-18, and GSDMD-N was reduced (P<0.05). The microarray and RT-PCR results showed that compared with BMSCs-Exo, the expression of LncRNA FGD5-AS1 in EPCs-Exo increased (P<0.05). Conclusion: EPCs-Exo inhibited paclitaxel-induced pyrolysis of vascular endothelial cells.
2022 Vol. 52 (1): 9-14 [Abstract] ( 467 ) HTML (1 KB)  PDF (1792 KB)  ( 660 )
15 Changes of kidney disease spectrum in Wenzhou area in recent 10 years: a retrospective study of 5 114 biopsy-proven cases
Lin Sishi, Chen Bo, Zhou Ying, You Xiaohan, Zhang Jianna, ChenXinxin, Ding Xiaokai, Zhang Ji, Chen Chaosheng
DOI: 10.3969/j.issn.2095-9400.2022.01.003
Objective: To analyze the composition and epidemiological trend of kidney disease spectrum in Wenzhou area in the past decade. Methods: A total of 5 114 biopsy-proven cases diagnosed in the Division of Kidney Pathology the First Affiliated Hospital of Wenzhou Medical University, from October 2009 to September 2019, were included in this study. They were divided on a 5-year basis as group 1 (October 2009 to September 2014) and group 2 (October 2014 to September 2019), with an analysis of the composition and evolution of kidney diseases. Results: The patients’ mean age at biopsy increased with time, and the proportion of the elderly patients
who received renal biopsy increased from 7.70% to 20.36%. Compared with group 1, group 2 had significantly increased the proportion of membranous nephropathy (MN) (24.3% vs. 36.8%, P<0.001), especially in the 45-59 age category. The secondary glomerular disease (SGD) mainly composed of immune-mediated diseases (55.0%),tumor-associated or metabolic diseases (33.8%) and infection-associated diseases (3.7%). Compared with group1, group 2 had nearly doubled proportion of tumor-associated or metabolic diseases (24.0% vs. 44.4%, P<0.001).Conclusion: In the past 10 years, the kidney disease spectrum in Wenzhou area has changed significantly. The proportion of MN was significantly increased and has become the most common PGD among middle-aged and elderly patients. In addition, the proportion of tumor-associated or metabolic diseases such as diabetic nephropathy and renal amyloidosis increased significantly.
2022 Vol. 52 (1): 15-22 [Abstract] ( 495 ) HTML (1 KB)  PDF (1681 KB)  ( 636 )
23 The association between dopamine D4 receptor 2-repeat allele and visual function network in children with attention deficit hyperactivity disorder
QIAN Andan, TAO Jiejie, CHEN Shuangli, ZHOU Ronghui, LOU
DOI: 10.3969/j.issn.2095-9400.2022.01.004
Objective: To investigate the effect of 48 bp 2-repeat allele (2R) in exon 3 of dopamine D4 receptor (DRD4) gene on visual function network in children with attention deficit hyperactivity disorder (ADHD) and its association with clinical symptoms. Methods: According to the DRD4 genotypes, 62 cases of noncarriers of 2R genotype and 41 cases of carriers of 2R genotype were divided into two groups. The visual level of continuous attention and response control were assessed by integrated visual and audio continuous performance test, and the severity of symptoms assessed by SNAP-IV. Three brain networks related to visual function were isolated by data-driven whole brain analysis, an independent component analysis, with further group analysis and inter-group comparison made thereafter. Results: The right temporal-occipital lobe of the lateral visual network of 2R carriers was prone to enhanced functional connectivity, while the right anterior central gyrus showed decreased connectivity and right anterior cingulate gyrus prone to decreased connectivity; the right superior occipital gyrus of the medial visual network of 2R carriers was prone to enhanced connectivity, while the left superior cerebellar gyrus prone to decreased connectivity. In the right temporal-occipital lobe, SNAP inattention scores were positively correlated with the intensity of functional connectivity in 2R carriers (r=0.339, P=0.037) and negatively correlated with the intensity of functional connectivity in 2R non-carriers (r=-0.308, P=0.016).Conclusion: DRD4 2R gene may promote the development of ADHD inattention symptom by mediating the pattern of visual network changes.
2022 Vol. 52 (1): 23-27,34 [Abstract] ( 592 ) HTML (1 KB)  PDF (1476 KB)  ( 9227 )
28 The inhibiting effect of osthole on proliferation and invasion of hepatocellular carcinoma cells
BAO Xiaodong, FANG Jie, JIN Junhua, ZHENG Guiru.
DOI: 10.3969/j.issn.2095-9400.2022.01.005

Objective: To investigate the effect of osthole on the proliferation and invasion of hepatocellular carcinoma cells HepG2, SMMC-7721, Bel-7402. Methods: Human liver cancer cells were taken for resuscitation,passage and culture. The experimental setting was divided into a blank group and an administration group. The administration group was treated with different concentrations of osthol liquid (50, 100, 150, 200 μmol/L). MTT method and cell colony experiment were used to determine the inhibitory effect of different concentrations of osthole on the proliferation of human liver cancer cells in vitro. Wound healing assay and transwell assay were used to investigate the inhibition of migration and invasion on HCC. Western blot was used to detect the influence of E-cadherin expression, a protein related to transformation (EMT) and MMP-2, a protein related to cell migration were detected by Western blot. Results: Osthole could inhibit the proliferation of HepG2, SMMC-77721 and Bel-7402 in vitro, with IC50 values of 147.62, 141.57 and 179.67 μmol/L, respectively, showing a certain concentrationdependent. Compared with the blank group, the treatment group could significantly inhibit the proliferation of human hepatocellular carcinoma cells under the conditions of 50, 75, and 100 μmol/L (P<0.05). It was found that osthole could effectively inhibit the migration and invasion of HCC. Compared with the blank group, the administration group showed statistically significant difference (P<0.05) The protein expression of MMP in the cells of the administration group decreased (P<0.05), while the expression of E-cadherin protein increased (P<0.05). Conclusion: Osthole can inhibit the proliferation and invasion in HCC and its mechanism may be related to protein E-cadherin and MMP-2.
2022 Vol. 52 (1): 28-34 [Abstract] ( 494 ) HTML (1 KB)  PDF (2115 KB)  ( 716 )
35 Establishment of risk prediction nomogram model for postpartum myofascial pelvic pain
CHEN Cong,NI Leyi, NI Feifei, CHEN Yumei.
DOI: 10.3969/j.issn.2095-9400.2022.01.006
Objective: To establish a risk prediction nomogram model for predicting the occurrence of postpartum pelvic myofascial pain. Methods: From July 2020 to September 2021, women who came to our hospital for follow-up visits 6-12 weeks after delivery, was assigned as MFPP group and non-MFPP group according to the diagnostic criteria of pelvic myofascial pain. General demographic data variables and pelvic floor EMG assessment data variables were screened by Lasso regression, and single-factor analysis was made for pelvic floor EMG assessment data. The screening variables were combined with the pelvic floor EMG assessment data commonly used in clinical diagnosis to construct the nomogram of the postpartum pelvic myofascial pain risk prediction model, and the calibration diagram was draw and adopt the consistency index (C-index) adopted to evaluate the stability and predictive power of the nomogram model. Then the clinical decision curve was drawn to analyze the quantification of the net benefit rate under different threshold probabilities in the postpartum MFPP cohort to determine the clinical applicability of the model. Results: A total of 300 postpartum followup women were collected (150 in the MFPP group and 150 in the non-MFPP group). Pregnancy, parity, delivery method, and newborn weight were related to the occurrence of postpartum MFPP. Postpartum MFPP risk prediction model nomogram C index was 0.706, with certain predictive ability. When the patient received a diagnosis threshold probability of 23%-72%, the net benefit rate of the model could be increased. Conclusion: The nomogram of the postpartum MFPP risk prediction model can effectively predict the postpartum MFPP and provide guidance for clinical evaluation.
2022 Vol. 52 (1): 35-40,46 [Abstract] ( 480 ) HTML (1 KB)  PDF (1555 KB)  ( 762 )
41 Observation of the effect of finger blocking exercises on the rehabilitation of adhesions after rupture of 2-5 finger flexor tendon in zone II
LIN Xiaoke, LIN Jingjing, NI Xiao, LI Haiyan.
DOI: 10.3969/j.issn.2095-9400.2022.01.007
Objective: To investigate the effect of blocking exercises on the rehabilitation of adhesions after rupture of 2-5 finger flexor tendon in zone II. Methods: Forty-three patients with adhesions after repairing the rupture of the flexor tendon zone II of the hand 2-5 fingers were selected and divided into 21 cases (33 fingers) in the observation group and 22 cases (36 fingers) in the control group according to the random number method. On the basis of conventional rehabilitation treatment, the observation group received finger blocking exercises, and the control group received conventional active finger flexion and extension training for 7 weeks. The total active motion (TAM) of the finger joints, the disability of arm shoulder and hand (DASH), and the pinch force of the affected finger to the thumb were evaluated and the comparison was calculated. Results: There was no significant difference between the two groups of patients in TAM and DASH before treatment. After treatment, the excellent and good rate of the observation group was 84.8% (28/33), and the excellent and good rate of the control group was 52.8% (19/36), the difference was statistically significant (χ2=8.757, P<0.05). Observation group TAM is 232.73°±28.87°, PIP is 74.36°±13.57°, DIP is 69.39°±15.20°, control group TAM is 192.75°±35.47°, PIP is 55.75°±17.13°, DIP is 47.78°±18.51°, the difference was statistically significant (P<0.05), the MP of the observation group was 88.97°±3.26°, and the MP of the control group was 89.22°±2.84°, the difference was not statistically significant (P>0.05). The DASH values of the observation group and the control group were lower than before treatment, and both were statistically significant (P<0.05). The DASH value of the observation group was lower than that of the control group, and the difference was statistically significant (P<0.05). The finger pinch force of the observation group was (6.85±2.12) Lbs, and the finger pinch force of the control group was (5.40±1.70) Lbs, and the difference was statistically significant (P<0.05). The percentage of finger pinch force in the observation group was (82.07±7.43)%, and the percentage of finger pinch force in the control group was (70.22±7.92)%. There were statistical differences between the two groups (t=6.400, P=0.001). Conclusion: Finger blocking exercises can significantly improve the active range of motion and finger pinching force of patients with adhesions after 2-5 finger flexor tendon rupture in zone II, and effectively improve the upper limb function of patients. The treatment effect is good.
2022 Vol. 52 (1): 41-46 [Abstract] ( 533 ) HTML (1 KB)  PDF (1469 KB)  ( 2257 )
47 The effect of anthracycline-induced cardiotoxity on left ventricular function in breast cancer patients: an evaluation with Tei index measured by tissue Doppler imaging
LUO Yinli, TAO Fangyi, CHEN Xiaowei,CHEN Shunping
DOI: 10.3969/j.issn.2095-9400.2022.01.008
Objective: To evaluate the effect of anthracycline-induced cardiotoxity on left ventricular function in breast cancer patients with Tei index measured by tissue Doppler imaging. Methods: Fifty-one cases of breast cancer treated with anthracycline-based chemotherapy were included in this study. According to the different chemotherapy cycles, the process consisted of 5 stages: the basic status before chemotherapy (T0), the end of the first chemotherapy cycle (T1), the end of the second chemotherapy cycle (T2), the end of the forth chemotherapy cycle (T3), and the end of the sixth chemotherapy cycle (T4). The conventional echocardiographic parameters and the Tei index were measured and compared. Results: There was no statistical difference in conventional echocardiographic parameters among the 5 stages (P>0.05), or in Tei index values between T0, T1 and T2 phases (P>0.05). However, Tei index value in T3 and T4 was significantly higher than in T0. Conclusion:Tei index is an earlier and more accurate assessment of cardiac toxicity caused by anthracycline-based chemotherapy in breast cancer patients, which is of high application value.
2022 Vol. 52 (1): 47-50,56 [Abstract] ( 535 ) HTML (1 KB)  PDF (1605 KB)  ( 680 )
51 Protective effect of Hirsutine on H2O2-induced oxidative stress injury in rat cardiomyocytes and its mechanisms
DING Lu, MA Luoping, MENG Mingxing, WANG Lei, JIANG Sheng, GUAN Zhimin,ZHANG Tingcai, LI Na
DOI: 10.3969/j.issn.2095-9400.2022.01.009
Objective: To investigate whether Hirsutine has protective effect on H2O2-induced oxidative stress injury in rat cardiomyocytes and its possible mechanisms. Methods: Primary neonatal rat cardiomyocytes,extracted from 1-2 days old neonatal rats, were stimulated with 400 μmol/L H2O2 to replicate the oxidative stress cell model. Cells were divided into 4 groups: Control, HS, H2O2, H2O2+HS. Cell activity, cell apoptosis and cell morphology were respectively detected by CCK8 assay, Tunel assay and immunofluorescence. Protein level of Bcl-2, Bax, Caspase-3 and Cleaved- caspase-3 was detected by Western blot. Results: It was found that the cell viability was significantly decreased after 6, 12, 24 hours of H2O2 treatment (P<0.01), compared with Control group, but it was significantly increased by 1 μmol/L and 10 μmol/L Hirsutine (P<0.01) compared with H2O2 group; Compared with H2O2 group, cell viability did not change and was markedly decreased by 100 μmol/L Hirsutine (P<0.01). Compared with H2O2 group, H2O2+HS group had significant increase in the apoptosis of cells (P<0.05) and the sarcomere structure was relatively intact. Compared with H2O2 group, the level of Bax/Bcl-2 and Cleaved-caspase 3/Caspase 3 was decreased by Hirsutine treatment, and Nrf2 and HO-1 protein increased by Hirsutine treatment (P<0.05). Conclusion: Hirsutine had protective effect on H2O2-induced mitochondrial apoptosis of rat cardiomyocytes, which may be mediated by Nrf2-HO-1 signaling pathway.
2022 Vol. 52 (1): 51-56 [Abstract] ( 589 ) HTML (1 KB)  PDF (1880 KB)  ( 668 )
57 The value of EBV DNA load in infectious mononucleosis
ZHANG Tingting, JIN Susu, CHEN Huale, LI Xiangyang
DOI: 10.3969/j.issn.2095-9400.2022.01.010
Objective: The present study aimed to evaluate the association between prognosis of disease of infectious mononucleosis (IM) with Epstein-Barr virus (EBV) DNA load in Wenzhou. Methods: A total of 1,155 children with IM were selected from the Second Affiliated Hospital & Yuying Children’s Hospital of Wenzhou Medical University from January 2018 to December 2020. Pertinent clinical data including patient age, gender,Length of hospitalization, percentage of abnormal lymphocytes and biochemical indexes included alanine aminotransferase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), EBV DNA and lymphocyte subsets were collected. The EBV DNA levels was divided into 4 groups (500-104, 104-105, 105-106, and >106 copies/mL). The Pearson correlation coefficient was used to show correlation. Results: Compare with the control group, ALT, LDH, Length of hospitalization, CD3+ T cells and CD8+ T cells in the EBV DNApositive group were higher (P<0.05), while CD4+ T cells and the ratio of CD4+/CD8+ T cells decreased (P<0.05).The ALT, LDH, Length of hospitalization, CD3+, CD8+ T cells in medium and high load groups were significantly higher than those in low load groups (P<0.05). There was a positive correlation between EBV DNA viral load and CD8+ T lymphocytes in patients with IM (r=0.156, P<0.05) and a negatively correlated with CD4+ T lymphocytes and the ratio of CD4+/CD8+ (r=-0.224, -0.213, P<0.05). Conclusion: EBV DNA load is related to the disease development and immune dysfunction in children with IM. Patients with higher EBV DNA load have more serious clinical symptoms and longer hospital stay, which should be paid more attention.
2022 Vol. 52 (1): 57-60 [Abstract] ( 512 ) HTML (1 KB)  PDF (1376 KB)  ( 653 )
61 Clinical evaluation of McNamara analysis for orthodontic and orthognathic combined surgical treatment in skeletal class III malocclusion
CHEN Hongyu, LAI Wenli
DOI: 10.3969/j.issn.2095-9400.2022.01.011
Objective: To evaluate the effect of McNamara analysis for orthodontic and orthognathic combined surgical treatment in skeletal class III malocclusion on adult patients. Methods: Twenty-one adult patients with skeletal class III malocclusion treated by orthodontic and orthognathic combined surgical treatment were randomly selected. Sagittal split ramus osteotomy and Le Fort I osteotomy were used in all orthognathic surgical treatment. Lateral cephalogram before and after treatment was evaluated through McNamara analysis by t test. Results: Maxillary length and maxillary protrusion increased, but mandibular length, mandibular protrusion,mandibular incisor protrusion and anterior lower facial height decreased after the treatment (P<0.05). Conclusion:After orthodontic and orthognathic combined surgical treatment, serious skeletal class III malocclusion was corrected in sagittal and vertical directions, and patient’s facial form was improved.
2022 Vol. 52 (1): 61-64 [Abstract] ( 455 ) HTML (1 KB)  PDF (1447 KB)  ( 886 )
65 Predictive analysis of sinus CT and peripheral blood cells for postoperative recurrence of inverted papilloma of nasal cavity and paranasal sinuses
XIE Rujiao, YOU Quanjie, HUANG Qi, REN Yuan, WU Zhenhua
DOI: 10.3969/j.issn.2095-9400.2022.01.012

Objective: To explore the clinical factors that affect the recurrence of sinonasal inverted papilloma, and to analyze the predictive value of sinus CT and peripheral blood cells for postoperative recurrence.Methods: A retrospective analysis was made of the clinical data of 38 SNIPs in the Ningbo Medical Center Lihuili Hospital (Eastern hospital compound) from January 2015 to December 2020, with clinical staging performed according to Krouse method. The results of CT staging and actual surgical staging were compared to study the accuracy of CT. At the same time, all cases were divided as recurrence group and non-recurrence group. Peripheral blood neutrophil count, lymphocyte count, neutrophil count/lymphocyte count and sinus CT score were counted, and demographic characteristics and other clinical parameters were compared between the two groups to study the predictive value of related factors of recurrence. Results: The actual coincidence rate of sinus CT in judging the scope of SNIP infringement was 92.11%. Lymphocyte count, NLR and sinus CT score were related to the postoperative recurrence of SNIP, of which NLR and CT score had higher predictive value.(AUC=0.813, 0.929, P<0.01). When NLR=2.375, the sensitivity was 81.8% and the specificity 81.5%; When CT score=5.5, the sensitivity was 81.8% and the specificity 96.3%. Conclusion: The accuracy of nasal sinus CT in judging the scope of SNIP violation is high, with high clinical utility. Lymphocyte count, NLR and sinus CT score are all related factors that affect the postoperative recurrence of SNIP, but NLR and CT score have higher predictive value.
2022 Vol. 52 (1): 65-68 [Abstract] ( 457 ) HTML (1 KB)  PDF (1398 KB)  ( 534 )
77 History, focus and future trend of research on internationalization strategy of higher education in China
ZHU Xuebo, JIANG Yuxi
DOI: 10.3969/j.issn.2095-9400.2022.01.016
Strategy plays an important role in the internationalization of higher education (IoHE). Using the Citespace visualization knowledge graph as the statistical analysis tool, and taking 304 papers from CSSCI Journals and Core Journals in CNKI database as samples, this paper discussed the history evolution, current focuses and future trend of IoHE in China. Research results showed that the history revolution of IoHE in China could be divided into four phases, while the contemporary focuses in this field could be mainly categorized into four types, namely, theoretical research on IoHE, learning the internationalization experience from counterparts overseas, experience sharing with domestic universities and colleges and studying IoHE strategy under a certain topic. This paper believes that we should improve the strategic research on IoHE from five aspects, that is, making in-depth theoretical research, keeping up with policy changes, joining university strategic alliances, conducting institutional research and defining value orientation.
2022 Vol. 52 (1): 77-82 [Abstract] ( 1244 ) HTML (1 KB)  PDF (1772 KB)  ( 1338 )
83 The sharing platform construction of large instrument in local universities
ZHOU Yiyang, GAO Hongchang
DOI: 10.3969/j.issn.2095-9400.2022.01.017
The large instrument sharing platform construction is an important part of country’s innovationdriven strategy and an important mission of higher education in the new era. As the main body of higher education, local universities should actively respond to the call of the country. The author analyzed the current situation and problems of the large instrument sharing platform construction in local universities by taking Wenzhou Medical University as an example and summarized the three innovative measures, i.e. laboratory safety “integrated” management, “sharing platform 3 plus” scaffold and platform technician team construction, with an aim to benefit the local university construction of large instrument sharing platform.
2022 Vol. 52 (1): 83-86,封三 [Abstract] ( 510 ) HTML (1 KB)  PDF (1498 KB)  ( 759 )
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