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2023 Vol. 53, No. 1 Published: 2023-01-25   1 Ectopic study of calcium phosphate cement seeded with antagomir-30d transfected BMSCs CHEN Wei, WANG Yan, JIN Qiong, HUANG Hui DOI: 10.3969/j.issn.2095-9400.2023.01.001 Objective: To constructed a compound of antagomir-30d/bone marrow mesenchymal stem cells (BMSCs)/calcium phosphate cement (CPC) and implant s.c. in BALB/c-nu mice to demonstrate that antagomir-30d could promote ectopic osteogenesis. Methods: The BMSCs transfected with the optimum concentration of antagomir-30d/NC/blank was loaded on calcium phosphate cement scaffolds and s.c. was implanted into BALB/c-nu mice. Implants were removed after 2, 4, 8 week (wk) respectively. 2-wk implants were subjected to RNA extraction, and then gene expression of osteoblast marker genes alkaline phosphatase (ALP), osteocalcin (OC) and Runt-related transcription factor 2 (RUNX2) was analyzed. 4-wk and 8-wk implants were subjected to histological staining and histomorphometric analysis was made to observe new bone formation. Results: Gene expression of osteoblast marker genes was analyzed after 2 wk implantation. RT-PCR analysis revealed up-regulation of ALP, OC and RUNX2 in the antagomir-30d-treated implants compared with implants transfected with miR negative control and the un-transfected group. The histological staining revealed that the antagomir-30d-treated implants had a small amount of new bone formation at 4 wk. Implants transfected with miR negative control and the un-transfected group showed new bone formation barely. Histomorphometric analysis showed that the percentage of new bone area of antagomir-30d-treated implants was 1.28%±0.19%. At 8 wk, new bone formation was clearly visible in all groups, while the new bone formation of antagomir-30d-treated group was more significant than NC and un-transfected groups (P<0.05). New bone area percentage was 17.79%±1.15%, 1.82%±0.53%, 2.33%±0.52% respectively. Conclusion: Antagomir-30d accelerates ectopic bone formation in vivo, and a compound of antagomir-30d/BMSCs/CPC is promising as tissue engineering complexes in repairing jaw bone defects. 2023 Vol. 53 (1): 1-6 [Abstract] ( 290 ) HTML (1 KB)  PDF (2069 KB)  ( 520 ) 7 Expression characteristics of interleukin-10 and its impact on psoriasis ZHENG Yi, PAN Lina, GAO Yu DOI: 10.3969/j.issn.2095-9400.2023.01.002 Objective: To analyze the characteristics of interleukin-10 (IL-10) in the skin lesions of psoriasis patients and psoriasis mouse models and its correlation with the severity of psoriasis, and to explore the effect of IL-10 expression on psoriasis. Methods: Samples of the skin lesions with psoriasis were collected from 40 patient between October 2018 and December 2019 in the Second Affiliated Hospital & Yuying Children’s Hospital of Wenzhou Medical University, while psoriasis-causing models were induced by imiquimod (IMQ) in C57BL/6 (WT) mice and IL-10 knockout (IL-10-/-) mice. PASI score was used to observe the dynamic changes of skin lesions. Morphological changes in tissues, epidermal thickness and degree of inflammatory cell infiltration were observed by HE staining. Immunohistochemical staining to analyze protein expressions of IL-10 and CD19 in skin lesions. Immunofluorescence staining was used to analyze the changes of CD4 localization in skin lesions. Results: IMQ model could have skin manifestations similar to clinical psoriasis. The erythema score, scale score, skin infiltration thickness score, PASI total score and ear thickness of mice were significantly higher in the IL-10-/- model group compared with WT modeling group and IL-10-/- control group (P<0.05). Immunohistochemical results showed that the positive expression rate of IL-10 in skin lesions of the WT model group was lower than that of the WT control group, with significant difference (P<0.05). The positive expression rate of CD19 in mice in IL-10-/- model group was slightly higher than that in IL-10-/- control group and WT model group, with significant difference (P<0.05). Immunofluorescence results showed that CD4 positive expression in WT model mice spread from the epidermal-dermal junction to the whole epidermis, similar to the CD4 positive localization in psoriasis patient samples. CD4 positive expression was present in both epidermis and dermis of IL-10-/- model mice. Conclusion: IL-10 deficiency leads to sustained immune activation and exacerbates the onset and progression of psoriasis, which may aggravate psoriasis-like lesion changes by increasing scaling occurrence and promoting the proliferation and differentiation of keratin-forming cells in the epidermal layer of the skin. 2023 Vol. 53 (1): 7-14 [Abstract] ( 397 ) HTML (1 KB)  PDF (2477 KB)  ( 618 ) 15 Establishment of the radiologic tumor invasion index based on radiomics splenic features and clinical factors to predict serous invasion of gastric cancer SHEN Ningzhe, ZHENG Jingwei, PAN Bujian, ZHANG Weiteng, CHEN Xiaodong DOI: 10.3969/j.issn.2095-9400.2023.01.003 Objective: To construct a model that can identify serosa infiltration in gastric cancer before surgery by combining clinical data and radiological features. Methods: A total 656 patients with pathologically confirmed gastric cancer were selected from the First Affiliated Hospital of Wenzhou Medical University between January 2015 and December 2019, who were randomly divided into the validation group (262 cases) and the modeling group (394 cases). The spleen imaging data of the patients in the modeling group were collected and analyzed by lasso regression, and the serosa infiltration prediction model was constructed on the selected significant features. Patients were divided into high-risk group (238 cases) and low-risk group (418 cases) by tumor infiltration risk score according to the cut-off value under the largest Yoden index, and then univariate and multivariate Logistic regression analysis was made with other infiltration-related factors to establish a visual prediction nomogram. Results: Patients with tumor infiltration score ≤-0.335 and > -0.335 were divided respectively into low-risk group and high-risk group. The diagnostic accuracy of the modeling and validation groups was consistent (P<0.001) when verified by the validation group (P<0.001). Univariate and multivariate Logistic analysis of infiltration risk factors showed that tumor radiomic infiltration score (OR=2.9, 95%CI=2.1-4.2, P<0.001), preoperative albumin (OR=1.3, 95%CI=1.2-3.1, P=0.003), platelet-lymphocyte ratio (OR=1.8, 95%CI=1.2-2.7, P=0.004) and tumor differentiation (OR=2.6, 95%CI=1.8-3.7, P<0.001) were independent influential factors for serosa invasion. The prediction model based on these four indicators accurately predicted the risk of serosa invasion, and its AUC value was 0.733. Conclusion: Tumor infiltration score based on spleen imaging combined with other clinical factors can accurately predict the serosal invasion of gastric cancer and improve the diagnostic precision. 2023 Vol. 53 (1): 15-21，28 [Abstract] ( 437 ) HTML (1 KB)  PDF (1667 KB)  ( 603 ) 22 A new AKT inhibitor induces apoptosisand autophagyin nasopharyngeal carcinoma cells ZHANG Jie, YANG Yufeng, HU Tingting, WU Cong, YANG Zifei, WU Xianmin DOI: 10.3969/j.issn.2095-9400.2023.01.004 Objective: To investigate the antitumor effect of E20, a novel AKT inhibitor, on human CNE-2 cells and the underlying regulatory mechanism. Methods: CCK8 assay was used to detect the activity of CNE-2 cells after E20 treatment. Cell apoptosis was detected by flow cytometry. Mito-sox were analyzed to detect the changes of mitochondrial ROS levels. Western blot was used to detect the expression of apoptosis-related proteins (AIF and Bcl-2) and autophagy-related proteins (LC3B-I, LC3B-II and P62) after E20 treatment. Transmission electron microscopy was used to observe the ultrastructure of mitochondria and the change of the autophagosomes. Results: E20 significantly inhibited the proliferation and promoted apoptosis of CNE-2 cells in a time-dose-dependent manner (P<0.05). After E20 treatment, the mitochondrial ROS level of CNE-2 cells was significantly increased, apoptosis protein AIF was significantly increased, and anti-apoptotic protein Bcl-2 was significantly decrease (all P<0.05). Autophagy-related protein LC3B-II was significantly increased, and P62 was significantly decreased (P<0.05). Transmission electron microscopy (TEM) showed that mitochondria were damaged and autophagosomes increased in E20 treated cells. Conclusion: E20 can inhibit the proliferation of NPC cells by inducing apoptosis through mitochondrial pathway and possibly activating autophagy, which provides a potential chemotherapy strategy for the treatment of NPC. 2023 Vol. 53 (1): 22-28 [Abstract] ( 305 ) HTML (1 KB)  PDF (2565 KB)  ( 573 ) 29 The role of miRNA-1 in cardiomyocyte pyroptosis and injury after cardiac arrest  cardiopulmonary resuscitation in rats YIN Jiana, LI Bingcan, ZHOU Peisen, LEI Yuanli, WANG Dongsheng, XU Huaqing, SONG Wenxing, HE Aiwen, LI Zhangping DOI: 10.3969/j.issn.2095-9400.2023.01.005 Objective: To investigate the role of miR-1 in myocardial pyroptosis and injury after cardiac arrest/cardiopulmonary resuscitation (CA/CPR) in rats. Methods: Cardiac arrest/cardiopulmonary resuscitation model was establishment and Spragu-Dawley (SD) rats were divided as cardiac arrest resuscitation (CA) group and sham operation control (Sham) group according to random number table method. The former was subdivided into 5 groups: CA group, CA+antago miR-1 group, CA+antago miR NC group, CA+ago miR-1 group, CA+ago miR NC group (n=6). All cardiac apical myocardial tissue and blood samples of all 6 groups were tested 6 hours after ROSC. The mRNA level of miRNA-1 in each group was detected using Real-time PCR. The expression of pyroptosis-related proteins including NLRP3, Caspase-1, IL-1β and IL-18 in each group was detected by Western blot. The pyroptosis rate of cardiomyocytes was detected by Tunel stain assay.The concentration of CK-MB and cTnI in serum was detected by ELISA. Results: There were no significant difference in body weight and basic MAP among the groups (P>0.05). Compared with Sham group, CA group had an increased expression of miR-1, NLRP3, Caspase-1, IL-1β, IL-18, pyroptosis rate, serum CK-MB and cTnI concentration (P<0.05). Compared with CA group, CA+antago miR-1 group had decreased expression of miR-1, NLRP3, Caspase-1, IL-1β, IL-18, pyroptosis rate, serum CK-MB and cTnI concentration (P<0.05); the expression of miR-1 and the expression of myocardial pyroptosis related proteins NLRP3, Caspase-1, IL-1β, IL-18, pyroptosis rate, serum CK-MB and cTnI concentration increased in CA+ago miR-1 group (P<0.05); there was no significant change in the expression of miR-1, NLRP3, Caspase-1, IL-1β, IL-18, pyroptosis rate, serum CK-MB and cTnI concentration in CA+antago miR NC group and CA+ago miR NC group (P>0.05). Conclusion: There was obvious myocardial pyroptosis and injury in CA/CPR rats after ROSC. Intravenous administration of ago miR-1 and antago miR-1 could regulate the expression of miR-1 in rat cardiomyocytes after CA/CPR, and miR-1 promoted myocardial pyroptosis and injury in rats after CA/CPR. The mechanism of miR-1 regulating myocardial injury after CA/CPR might be related to its promotion of myocardial pyroptosis. 2023 Vol. 53 (1): 29-35，41 [Abstract] ( 321 ) HTML (1 KB)  PDF (2283 KB)  ( 631 ) 36 The sensitizing effect of targeting liver X receptor on PD-1 monoclonal antibody in the treatment of primary liver cancer ZHOU Yu, ZHANG Weiwei, GAO Youting DOI: 10.3969/j.issn.2095-9400.2023.01.006 Objective: To investigate the correlation between liver X receptor (LXR) and programmed death-ligand 1 (PD-L1) expression and the sensitizing effect of the agonist of LXR (T09) on the therapeutic effect PD1 monoclonal antibody against primary hepatocellular carcinoma (HCC). Methods: Bioinformatics analysis was applied in the present study to identify the correlation between the expression of LXR associated with its downstream target ABCA1 and the expression of PD-L1 in HCC. In addition, the protein level correlation between LXR and PD-L1 in liver cancer tissue samples was examined by immunohistochemical staining. The impact of hepatocellular carcinoma cells-derived LXR on PD-L1 expression was identify by knockdown of LXR using lentivirus-mediated siRNA. In vivo study, the orthotopic liver cancer transplantation mice model (Hepa1-6) mice were treated with T09 and (or) PD1 monoclonal antibody, and the subcutaneous tumor volume was measured after 3-week- intervention. Results: Bioinformatic analysis showed that the expression of LXR and ABCA1 was significantly correlated with the expression of PD-L1 (correlation co-efficient r=0.12, P=0.021; Correlation co-efficient between LXR expression and PD-L1 expression, r=0.20, P<0.001). Immunohistochemical staining showed that both the expression levels of LXR and PD-L1 were significantly increased in tumor tissues. Patients with high expression of LXR were often associated with high expression of PD-L1. Immunohistochemical Chi-square test analysis showed that the expression of PD-L1 and LXR was correlated, with significant difference (P<0.05). Additionally, knockdown of LXR expression in HCC cell lines was associated with the decreased PD-L1 expression. In vivo study, The co-treatment strategy significantly enhanced the anti-tumor effect of single PD1 monotherapy treatment strategy. Conclusion: The expression of LXR was significantly correlated with PD-L1, and the agonist of LXR could significantly increase the sensitivity of PD1 in the treatment of liver cancer. 2023 Vol. 53 (1): 36-41 [Abstract] ( 302 ) HTML (1 KB)  PDF (1877 KB)  ( 736 ) 42 Protective effect of butyrate on renal injury in diabetic nephropathy mice and its mechanism YE Kaili, HUANG Shiqin, HU Ting, ZHAO Yanling DOI: 10.3969/j.issn.2095-9400.2023.01.007 Objective: To investigate the protective effect of butyrate on renal injury in mice with type 2 diabetic nephropathy and to explore its mechanism. Methods: Male db/db mice were randomly divided into model group (DN), sodium butyrate 500 mg·kg-1·d-1 group (NaB1) and sodium butyrate 1 000 mg·kg-1·d-1 group (NaB2). The db/m mice with the same weeks served as the control group (NC). NaB1 and NaB2 groups were given sodium butyrate solution every day, while NC group and DN group were perfused with equal volume of physiological saline. After 8 weeks for intragastric administration, the mice were sacrificed. Kidney tissue was preserved for staining of hematoxylin eosin (HE) and periodic acid-Schiff (PAS); meanwhile extracting the mRNA and protein for real-time quantitative polymerase chain reaction (RT-qPCR) and western blotting; using relevant kits to detect urine creatinine, urine microalbumin (mALB), interleukin-6 (IL-6) of renal tissue, serum creatinine (SCr) and other related indicators. Results: Compared with the NC group, the DN group presented listlessness, more polydipsia and polyuria, expressed significantly higher body weight and blood glucose (P<0.05), and increased the ratio of urinary protein and creatinine (UACR) (P<0.05). After the treatment of sodium butyrate for 8 weeks, the mice had better mental conditions and diet, lower blood glucose and UACR (P<0.05) and lower IL-6 levels of renal tissue, but higher expressions of GLP-1R, AMPK, PGC-1α, MFN2 and OPA1 mRNA (P<0.05). Conclusion: Sodium butyrate exerts reno-protective effect by regulating AMPK and GLP-1R in diabetic nephropathy mice. 2023 Vol. 53 (1): 42-48 [Abstract] ( 538 ) HTML (1 KB)  PDF (2254 KB)  ( 677 ) 49 The value of two-dimensional speckle tracking imaging for predicting MACE in AMI patients one year after PCI WENG Wenchao, YANG Daoling, CHEN Jingwan DOI: 10.3969/j.issn.2095-9400.2023.01.008 Objective: To evaluate two-dimensional ultrasound speckle tracking imaging (STI) in predicting major adverse cardiac events (MACE) for acute myocardial infarction (AMI) patients with one-year follow-up after percutaneous coronary intervention (PCI). Methods: From January 2019 to July 2021, 91 patients with AMI for PCI into Jinhua Municipal Central hospital were included in the study. They all received routine ultrasound and two-dimensional STI one week after surgery and were all followed up for 1 year, with MACE recorded (mainly including recurrent angina pectoris, recurrent myocardial infarction, new heart failure, malignant arrhythmia, rehospitalization and sudden cardiac death). Results: A total of 18 cases (19.78%, 18/91) with MACE were followed up, the time of MACE being 12-46 weeks after discharge, with median time of 37.5 weeks. Patients were divided into MACE group and non-MACE group, with no differences (P>0.05) found in gender, age, AMI type, PCI mode, target lesion stenosis rate, number and length of implanted stents, and postoperative medicine treatments. The left ventricular ejection fraction (LVEF) in MACE group was significantly lower and serum N-terminal pro-brain natriuretic peptide (pro-BNP) was significantly higher than that in non-MACE group (P<0.05); however, there were no differences in left ventricular end diastolic diameter and volume, end systolic diameter and volume, serum troponin I (cTnI) and creatine kinase isoenzyme (CK-MB) between the two groups (P>0.05). STI parameters including global longitudinal strain (GLS) and global circumferential strain (GCS) in MACE group were significantly higher than those in non-MACE group and global radial strain (GRS) was less too (P<0.05). Multivariate logistic regression model found that age≥70-years-old (OR=1.652, 95%CI=1.234-1.896, P=0.003), LVEF≤40% (OR=3.021, 95%CI=2.456-3.859, P<0.001), pro-BNP≥500 ng/mL (OR=2.235, 95%CI=1.652-3.002, P<0.001), GLS≥-22% (OR=1.958, 95%CI=1.452-2.639, P<0.001), GRS≤44% (OR=1.325, 95%CI= 1.084-1.996, P=0.005) and GCS≥-16% (OR=1.421, 95%CI=1.162-2.032, P=0.003) were all the independent risk factors of MACE. The receiver operating curve (ROC) showed that the area under the curve (AUC) of GLS, GRS and GCS for predicting MACE were 0.842, 0.803 and 0.756, respectively. Kaplan-Meier survival curve showed that the cumulative MACE incidence of patients with GLS≥-22% was significantly higher than patients with GLS < -22%, patients with GRS≤44% was significantly higher than patients with GRS > 44%, and patients with GCS≥-16% was significantly higher than patients with GCS < -16% (P<0.05). Conclusion: In addition to the conventional indicators such as age, LVEF and pro-BNP, two-dimensional ultrasound STI parameters GLS, GRS and GCS is also of great value for predicting the occurrence of MACE for AMI patients one year after PCI. 2023 Vol. 53 (1): 49-54 [Abstract] ( 332 ) HTML (1 KB)  PDF (1502 KB)  ( 499 ) 55 A comparative study between the laparoscopic intraperitoneal onlay mesh and the totally visceral sac separation approach in ventral hernia ZHUANG Bo, YU Kai, ZHANG Zhiwei, GONG Daojun, YU Shi’an DOI: 10.3969/j.issn.2095-9400.2023.01.009 Objective: To investigate the difference between the technique of totally visceral sac separation (TVS) and the laparoscopic intraperitoneal onlay mesh (IPOM) in ventral hernia from the perspective of the clinical application of the TVS procedure and the evaluation of its safety, efficacy and cost-effectiveness. Methods: Data of 56 patients who underwent ventral hernia in Department of General Surgery Jinhua Hospital Zhejiang University School of Medicine from October 2019 to October 2021 were retrospectively analyzed, among whom 27 patients received TVS procedures (TVS group) and 29 patients IPOM operations (IPOM group). Baseline characteristics, operative records and perioperative data were compared between two groups. Follow-up was conducted by outpatient examination and telephone interview to detect patients’ survival and recurrence of ventral hernia. Results: There were no differences in age, sex, BMI,EHS- classification and defect area between two groups (P>0.05). Operative time in TVS procedure was significantly longer with mean time of (216.93± 52.80) min and (187.17±40.73) min in IPOM patients (P=0.021). The visual analogue score (VAS) on first postoperative day and hospitalization cost of the TVS group was lower than that of IPOM group (P=0.003, P<0.001). There was no significant difference in postoperative hospital stay (P=0.067). All procedures were successful, but two TVS group patients underwent open conversion, and one IPOM group patient had a surgical site infection that was cured conservatively. There was one case of recurrence after 6 months in TVS group and no perioperative mortality was found. The patients had been followed-up until March 2022, with the mean postoperative follow-up time being 14 months and the follow-up rate 94.6%. Conclusion: The TVS procedure is time-consuming but safe and feasible. It is a low-cost supplementary technique for surgical ventral hernias. 2023 Vol. 53 (1): 55-59 [Abstract] ( 296 ) HTML (1 KB)  PDF (1868 KB)  ( 500 ) 60 Characteristic factors of unplanned endotracheal extubation events in intensive care unit: an analysis based on association rules HAN Yongqi, HU Qingwen, CHEN Hao, LI Xueyan, XU Xiaoqun DOI: 10.3969/j.issn.2095-9400.2023.01.010 Objective: To explore the related factors for unplanned endotracheal extubation events in intensive care unit (ICU), so as to provide scientific basis for the prevention and management of unplanned endotracheal extubations in ICU patients. Methods: Based on the association rules, data of 109 cases of unplanned endotracheal extubations reported from the First Affiliated Hospital of Wenzhou Medical University between January 1st 2018 to August 31st 2021 were analyzed. Results: Through condition setting and Chi-square test, 309 rules were selected. With nursing professional knowledge taken into account, 16 strong association rules were finally obtained. Conclusion: Through data mining, we can see clearly the  association rules among factors of unplanned endotracheal extubation events, which provides guidance for the prevention and management system of unplanned endotrachea extubations in ICU, thus ensuring the safety of patients with endotracheal intubation. 2023 Vol. 53 (1): 60-66 [Abstract] ( 355 ) HTML (1 KB)  PDF (1367 KB)  ( 726 ) 79 Enhancement of teachers’ professional ethics in colleges and universities in the new era: a four dimensional analysis YE Xiaoli, LI Junhong DOI: 10.3969/j.issn.2095-9400.2023.01.015 Teachers’ professional ethics are the first and foremost criteria in evaluating teachers. Since the 18th National Congress of the Communist Party of China, General Secretary Xi Jinping has given a series of important instructions, as well as personally deployed the cultivation of teachers’ professional ethics in colleges and universities. However, while the overall situation of teachers’ moral cultivation currently shows a positive tendency, there still exist some problems to be addressed with regard to the synergy management mechanism,teachers’ educational consciousness, its effectiveness, and the evaluation or assessment system concerned. Standing at the new historical juncture, colleges and universities are supposed to focus on the new situation and further strengthen the cultivation of teachers’ ethics in the new era, thus comprehensively enhancing the teachers’ ethics in colleges and universities through building an operating system, a leading system, an assessment system, and a caring system. 2023 Vol. 53 (1): 79-83 [Abstract] ( 298 ) HTML (1 KB)  PDF (1330 KB)  ( 958 ) 84 The present situation and countermeasures of medical students returning to their homes for employment in the post epidemic period——A case study of 1 078 medical graduates from 90 counties and districts in Zhejiang province ZHENG Yanfei, WANG Zhangye, HUANG Zhifeng DOI: 10.3969/j.issn.2095-9400.2023.01.016 Altogether 1 078 medical graduates from 90 counties and districts in Zhejiang Province were interview and investigated through questionnaire to explore the current situation, influencing factors and countermeasures of medical students returning to their hometown for employment in the post-pandemic period. The results showed that the willingness of medical students to return to their hometown for employment was above the middle level, and the job intention was relatively concentrated. Male students, students from eastern Zhejiang, the only child family, and students with social practice and entrepreneurial experience were more willing to return home for employment. The influencing factors included policy orientation, family education, welfare benefits, income level, social security, and nostalgia for hometown. Accordingly, the author puts forward the countermeasures to construct the social support for returning medical students’ employment, namely: strengthening the governments’ regulation and improving education and welfare benefits for the children of returned graduates. We should change medical students in their employment concept, reconstruct their “native emotion”, and improve their services, dedication and governance capability for rural areas. 2023 Vol. 53 (1): 84-86，封三 [Abstract] ( 441 ) HTML (1 KB)  PDF (1346 KB)  ( 756 )

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