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28 August 2021, Volume 41 Issue 8 Previous Issue   
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Innovative research team achievement column
Basic research
Clinical research
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Innovative research team achievement column

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Invitro screening and validation of novel targeted therapeutic strategy against diffuse intrinsic pontine glioma Rui LI, Yu-jie HAN, Lei ZHANG, Yu-jie TANG 2021, 41 (8):  987-998.  doi: 10.3969/j.issn.1674-8115.2021.08.001 Abstract ( 608 )   HTML ( 29 )   PDF (13331KB) ( 300 )   ·To find and identify new targeted therapeutic compounds and combinations for diffuse intrinsic pontine glioma (DIPG) from the perspective of epigenetics. ·Selection of small molecule compounds was based on the previously published transcriptome analysis of 8 cases of DIPG and 6 cases of normal brain tissues. New inhibitory compounds of DIPG were identified by single agent screening in DIPG primary tumor cells. The changes of target genes at mRNA and protein expression level were detected by real-time PCR and Western blotting after drug treatment. The effects of drug treatment on the proliferation and apoptosis of DIPG primary tumor cells were detected by FACS analyses after EdU and Annexin V/propidium iodide staining, respectively. The combinatory screening of small molecular compounds was performed with bromodomain and extra terminal protein (BET) inhibitor JQ1 or histone deacetylase (HDAC) inhibitor panobinostat, and the drug combination with inhibitory effect on DIPG was verified in vitro. ·Sixty-six small molecules were chosen to be applied to screening. Single agent screening identified that YM155 could significantly inhibit DIPG primary tumor cell growth, and BIRC5 (baculoviral IAP repeat containing 5; gene encoding survivin), a target gene of YM155, was significantly upregulated in DIPG tumor tissues (P=0.018). YM155 could reduce the expression of BICR5 at both mRNA and protein levels. YM155 could repress proliferation and induce apoptosis of DIPG. CX4945 and ABT-737 from the targeted small molecular library were combined with JQ1 (BET inhibitor) and panobinostat (HDAC inhibitor), respectively,which could synergistically inhibit the activity of DIPG cells in vitro. ·Novel targeted therapeutic strategies for DIPG has been identified through single drug and combination drug screening, providing basis for furthervalidation in vivo and therapeutic mechanism exploration. Figures and Tables | References | Related Articles | Metrics

Basic research

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Haptoglobin suppresses hepatocyte ferroptosis via inhibition of the ERK1/2 signaling pathway Qian WEI, Ying-ting ZHANG, Long-shuai LIN, En-jun HE, Yong-yuan HE, Ying-hong SU, Cheng-cheng DUAN, Si-yuan WANG, Qing-hua ZHAO, Qian ZHAO, Ming HE 2021, 41 (8):  999-1008.  doi: 10.3969/j.issn.1674-8115.2021.08.002 Abstract ( 1085 )   HTML ( 96 )   PDF (4390KB) ( 911 )   ·To explore the role and mechanism of haptoglobin (Hp) in hepatocyte ferroptosis. ·Thirteen 8-week-old C57BL/6J male mice were randomly divided into two groups: one group of mice were fed with normal iron diet (NID group, n=5) and the other were fed with high iron diet (HID group, n=8). After 12 weeks, the murine serums and livers were collected from both groups. Liver injury, histopathological changes, hepatic fibrosis, iron deposition and peroxidation in murine liver tissues were determined by serum index test, hematoxylin-eosin (H-E) staining, Sirius red staining, Prussian blue staining and 4-hydroxynonenal (4-HNE) staining, respectively. RNA-sequencing (RNA-seq) and bioinformatics were used to analyze the effect of high iron diet on the expression of transcriptome in the murine livers and to screen new candidate genes that might regulate ferroptosis. After Hp was overexpressed or knocked down in mouse liver cells (AML-12), RAS-selective lethal small molecule 3 (RSL3) or/and extracellular regulated protein kinases 1/2 (ERK1/2) inhibitor SCH772984 was/were added to cells. The role and mechanism of Hp in hepatocyte ferroptosis were determined by cell counting, real-time PCR, C11-BODIPY581/591 immunofluorescence and flow cytometry. ·After 12 weeks of feeding, the levels of glutamic-pyruvic transaminase (GPT) and glutamic-oxalacetic transaminase (GOT) in the HID group significantly increased compared with the NID group. The high iron diet induced hepatocytes death, hepatic fibrosis, iron deposition and lipid peroxidation products accumulation. RNA-seq analysis and bioinformatics results showed that acute phase response, catalytic activity response, antioxidant activity and hemoglobin binding signaling pathways were significantly regulated in the livers of HID mice, and Hp mRNA decreased significantly. Furthermore, the mRNA and protein expression levels of Hp in liver tissues of HID mice and in AML-12 hepatocytes treated with RSL3 obviously decreased. Importantly, Hp significantly alleviated lipid peroxidation and ferroptosis induced by RSL3 in AML-12 hepatocytes. Meanwhile, Hp suppressed RSL3-induced phosphorylation of ERK1/2 in AML-12 hepatocytes. Moreover, ERK1/2 inhibitor SCH772984 totally reversed the aggravation of hepatocyte ferroptosis induced by Hp knockdown. ·Hp is a novel inhibitory molecule in hepatocytes ferroptosis, which alleviates hepatocyte ferroptosis and liver injury by inhibiting ERK1/2 signaling pathway. Figures and Tables | References | Related Articles | Metrics

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Expression of protein arginine methyltransferase 5 in lung cancer and its mechanism of promoting lung cancer Bing-qian ZHOU, Li HAN, Zhe-yi CHEN, Shi-yu CHEN, Ying-xia ZHENG 2021, 41 (8):  1009-1016.  doi: 10.3969/j.issn.1674-8115.2021.08.003 Abstract ( 611 )   HTML ( 87 )   PDF (3871KB) ( 503 )   ·To investigate the expression of protein arginine methyltransferase 5 (PRMT5) in lung cancer and its effect on proliferation, migration and invasion of lung cancer cells. · Immunohistochemistry was used to detect the expression of PRMT5 in 73 pairs of lung cancer tissues and adjacent tissues, and its correlation with clinical characteristics of patients was analyzed. After PRMT5 was down-regulated in lung cancer cell line NCI-H1299, the effects of PRMT5 on cell proliferation, migration and invasion were observed by CCK8 assay and Transwell chamber, and the expressions of E-cadherin, vimentin and transcription factor SNAI1 were detected by Western blotting. The expression of PRMT5 in tumor cell lines and lung cancer tissues was analyzed by using the Gene Expression Omnibus (GEO) and The Cancer Genome Atlas (TCGA) database, respectively, and the relationship between the expression of PRMT5 in tissues and the prognosis of lung cancer patients was further analyzed. ·Immunohistochemistry showed that the expression of PRMT5 in lung cancer tissues was significantly higher than that in adjacent tissues (P=0.000), and the expression of PRMT5 in stage Ⅲ?Ⅳ was higher than that in stage Ⅰ?Ⅱ (P=0.033). After PRMT5 down-regulation, the proliferation, migration and invasion ability of NCI-H1299 cells decreased, the expression of E-cadherin increased and the expression of vimentin and SNAI1 decreased. The lung cancer sequencing data of TCGA database showed that the expression of PRMT5 in lung cancer tissues was significantly higher than that in normal lung tissues, and the lung adenocarcinoma patients and the lung squamous cell carcinoma patients with higher expression of PRMT5 had lower overall survival rate (P=0.005) and relapse-free survival rate (P=0.028), respectively. · PRMT5 is highly expressed in lung cancer tissues, which can promote the proliferation, migration and invasion of lung cancer cells. High expression of PRMT5 is associated with poor prognosis of lung cancer. Figures and Tables | References | Related Articles | Metrics

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Establishment and optimization of co-culture technology for breast cancer organoids Tian-hao ZHOU, Zhao-chen XIN, Shao-qian DU, Yuan CAO, Jing-xuan XU, Zeng-hong LAO, Hong-xia WANG 2021, 41 (8):  1017-1024.  doi: 10.3969/j.issn.1674-8115.2021.08.004 Abstract ( 1611 )   HTML ( 234 )   PDF (4693KB) ( 1178 )   ·To improve the cultivating and passaging method of breast cancer organoids, and establish a co-culture system enriching cancer associated fibroblasts (CAFs). ·Different types of collagenases (type Ⅰ, type Ⅲ and type Ⅳ) were used to digest fresh tissues from 5 breast cancer patients. The number of cells after tissue digestion was counted by cell counting method, and cell viability was analyzed by cell flow cytometry. Three-dimensional culture of primary breast cancer single cells was carried out by using culture system containing different contents of fibroblast growth factor 7 (FGF7), FGF10 and epidermal growth factor (EGF). The success rate of cell culture and the growing status of organoids were observed and compared. Different centrifugation speeds were used to compare the advantages and disadvantages of passaging methods and simplify the passaging steps. CCK8 assay was used to study the effect of CAFs on the growth of organoids in the co-culture system of primary CAFs and organoids, and the morphological changes of organoids were observed under optical microscope. ·Compared with type Ⅰ and type Ⅲ collagenase, type Ⅳ collagenase got the highest cell yields (P=0.045, P=0.017), and maintained the highest cell viability (P=0.005, P=0.048). By optimizing the composition of organoid medium (omitting FGF7 and FGF10, reducing EGF concentration) and passaging process (improving centrifugal velocity to 900×g), a more economical, effective and rapid method of organoid culture was obtained. Compared with organoids cultured alone, the growth rate (P<0.05) and heterogeneity of organoids increased when organoids were co-cultured with CAFs. ·The optimized culture system can significantly increase the success rate of organoids, simplify the culture steps and reduce the culture cost. The establishment of primary CAFs and organoids co-culture system provides a good in vitro model for the study of breast cancer heterogeneity and tumor microenvironment. Figures and Tables | References | Related Articles | Metrics

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Inhibition of CDDO-ME on ubiquitin-specific protease 2a activity and cell proliferation in triple negative breast cancer cells Yan-jie JI, Hao LUO, Hai-yan CAI, Xin-yu LIU, Shi-jia JIN, Shen-yue SU, Han-zhang XU, Hu LEI, Ying-li WU 2021, 41 (8):  1025-1032.  doi: 10.3969/j.issn.1674-8115.2021.08.005 Abstract ( 640 )   HTML ( 38 )   PDF (2591KB) ( 378 )   ·To explore the effect of bardoxolone methyl (CDDO-Me), an inhibitor of ubiquitin-specific protease 2a (USP2a) screened in vitro, on USP2a activity and cell proliferation in the triple negative breast cancer (TNBC) cells. ·Ubiquitin-specific protease inhibitor screening system was used to screen USP2a inhibitors and CDDO-Me was obtained. Molecular docking technology was used to analyze the interaction of CDDO-Me and USP2a. Cellular thermal shift assay (CETSA) was used to detect the interaction between CDDO-Me and USP2a protein in three TNBC cell lines. Western blotting was used to detect the changes of USP2a substrates including β-catenin and tumor necrosis factor receptor-associated factor 6 (TRAF6) protein levels and apoptosis-related proteins including caspase3 and poly (ADP-ribose) polymerase 1 (PARP1). Cell counting kit-8 (CCK8) was used to detect the effect of CDDO-Me on the proliferation of TNBC cells. MDA-MB-468 cells were transiently transfected with pLVX (pLVX group) or pLVX-USP2a (pLVX-USP2a group) plasmids. After CDDO-Me treatment, the protein levels of β-catenin and TRAF6 were detected by Western blotting, the cell cycle was detected by flow cytometry, and the number of viable cells was detected by trypan blue exclusion method. ·CDDO-Me inhibited the activity of USP2a in vitro, and half-maximal inhibitory concentration was 3.84 μmol/L. The results of molecular docking analysis showed that CDDO-Me formed a hydrogen bond with His456 residue of USP2a, and had hydrophobic interactions with Phe409 and Tyr514 residues. CETSA results showed that CDDO-Me binded to the USP2a protein in the three TNBC cells. The results of Western blotting showed that CDDO-Me down-regulated the protein levels of β-catenin and TRAF6, while the two USP2a substrates did not decrease in the USP2a-overexpressed MDA-MB-468 cells treated by the same concentration of CDDO-Me. CDDO-Me inhibited the proliferation of TNBC cells in a dose-dependent manner, caused caspase3 activation and PARP1 cleavage, and led to S phase and G2/M phase arrest. Compared with the pLVX group, there were more viable cells in the pLVX-USP2a group and the cells also did not undergo cycle arrest. ·CDDO-Me can inhibit the activity of USP2a in TNBC cells, inhibit the proliferation of TNBC cells and induce apoptosis. Figures and Tables | References | Related Articles | Metrics

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Expression and prognosis analysis of MRPL12 in lung adenocarcinoma Lei XIONG, Qian YI, Ming-fang XU, Jian CHEN 2021, 41 (8):  1033-1040.  doi: 10.3969/j.issn.1674-8115.2021.08.006 Abstract ( 992 )   HTML ( 109 )   PDF (4968KB) ( 671 )   ·To analyze the difference of expression of mitochondrial ribosomal protein L12 (MRPL12) in lung adenocarcinoma (LUAD) and normal tissues and its influence on the prognosis of patients, and predict its biological function. ·The expressions of MRPL12 in different cancer tissues were analyzed by The Cancer Genome Atlas (TCGA) using the tumor-infiltrating immune cell analysis database TIMER and GEPIA (gene expression profiling and interactive analyses) server. The expressions of MRPL12 in different types of cancer were analyzed by Sangerbox bioinformatics platform. Cox regression and Kaplan-Meier survival analysis were used and the hazard ratios (HR), the 95% confidence intervals (CI) and the Log-rank P values were evaluated. The LinkedOmics database was used to identify the genes associated with MRPL12 expression. The DAVID database was used for the enrichment analysis of Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway. The transcription levels of MRPL12 in cells were verified by quantitative PCR (qPCR). ·MRPL12 was highly expressed in many cancer types. But only the LUAD patients with high MRPL12 expression had lower overall survival (OS) and disease-specific survival (DSS). The expression of MRPL12 protein increased with tumor stage and individual tumor grade. The genes negatively related to MRPL12 were enriched into several cancer-related pathways in LUAD gene expression data set, including pathways in cancer (P=0.000), proteoglycans in cancer (P=0.000) and non-small cell lung cancer (P=0.000). The mRNA expression levels of MRPL12 in the LUAD cells (H1395, H1975 and HCC827) were higher than that in the normal cell (Beas-2B), and the difference was statistically significant (P<0.05). ·The LUAD patients with high expression of MRPL12 may have poor prognosis, and thus MRPL12 can be a potential biomarker and potential therapeutic target for LUAD. Figures and Tables | References | Related Articles | Metrics

Clinical research

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Comparison of mismatch negativity in patients with schizophrenia and depression Meng-ting SHEN, Xuan-hong ZHANG, Zhen-ying QIAN, Hui LI, Jian-hua SHENG, Ji-jun WANG, Ying-ying TANG 2021, 41 (8):  1041-1045.  doi: 10.3969/j.issn.1674-8115.2021.08.007 Abstract ( 812 )   HTML ( 14 )   PDF (1250KB) ( 478 )   ·To compare the difference of mismatch negativity (MMN) between first-episode schizophrenia and depression and its correlation with clinical characteristics, and explore the role of pre-attention processing in the mechanisms of schizophrenia and depression. ·Twenty patients with schizophrenia and 19 patients with depression were selected in the outpatient department of Shanghai Mental Health Center, Shanghai Jiao Tong University School of Medicine from January 2014 to December 2016. Healthy controls were also recruited. The Positive and Negative Syndrome Scale (PANSS) was used to measure the clinical symptoms of schizophrenia. Hamilton Depression Scale (HAMD) and Hamilton Anxiety Scale (HAMA) were used to measure the clinical symptoms of depression. All patients and healthy controls completed MMN detection and clinical scale assessments. The amplitudes and latencies of duration MMN (DMMN) and frequency MMN (FMMN) were analyzed by repeated measures analysis of variance with group as the between-group factors. The midline electrodes (Fz, Fcz) were used as the with-in group factor; meanwhile, region (F, Fc) and laterality (1 for left, 2 for right) were used as the within-group factors for lateral electrodes and the factor sex was used as covariate. Partial correlations was performed to find the correlation between MMN and clinical characteristics. ·① The amplitude of DMMN in patients with schizophrenia 2 for [(-2.70±2.46) μV)] was lower than that in patients with depression [(-5.06±0.46) μV] and healthy controls [(-5.15±0.43) μV] (both P≤0.001). No significant group differences of DMMN latency were observed at midline or lateral electrodes (All P>0.05). ② There was no significant group difference of FMMN amplitudes at midline electrodes (P>0.05) but a significant between-group difference at lateral electrodes (P=0.040). No significant group differences of FMMN latency were observed at midline or lateral electrodes (All P>0.05). ③ There was a significant correlation between DMMN amplitudes at F2 (P=0.042) or Fz (P=0.032) and general scores of PANSS in schizophrenia patients. There was no correlation in depression patients. ·Abnormality of DMMN exists in the patients with first episode schizophrenia but not in the patients with depression, suggesting that DMMN amplitudes may work as a biological marker to distinguish schizophrenia and depression. Figures and Tables | References | Related Articles | Metrics

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Application of adjusted global antiphospholipid syndrome score to risk assessment of new thromboembolic events in patients with systemic lupus erythematosus Jia LI, Ran WANG, Chun-de BAO, Nan SHEN, Qiong FU 2021, 41 (8):  1046-1050.  doi: 10.3969/j.issn.1674-8115.2021.08.008 Abstract ( 576 )   HTML ( 7 )   PDF (1099KB) ( 444 )   ·To verify the efficacy of adjusted global antiphospholipid syndrome score (aGAPSS) in the risk assessment of new thromboembolic events in the patients with systemic lupus erythematosus (SLE). ·A total of 330 SLE patients admitted to Renji Hospital, Shanghai Jiao Tong University School of Medicine from December 2012 to March 2016 were retrospectively analyzed. The patients were divided into thrombosis group and control group according to whether there were new thromboembolic events during hospitalization. Demographic data and clinical data of the two groups were collected, and aGAPSS was calculated according to the five risk indicators. Receiver operator characteristic curve (ROC curve) was used to evaluate the diagnostic efficacy of aGAPSS in the patients with new thromboembolic events. ·A total of 60 patients (18.2%) had new thromboembolic events. Compared with the control group, aGAPSS significantly increased in the thrombosis group (P=0.000). The results of ROC curve showed that when the aGAPSS>7 scores, the area under the curve for the diagnosis of new thromboembolic events was 0.752 (95%CI 0.702-0.797), and the sensitivity and specificity were 48.33% and 94.07%, respectively. ·aGAPSS can be used to assess the risk of thromboembolic events in SLE patients and provide reference for clinical treatment decision-making. Figures and Tables | References | Related Articles | Metrics

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Analysis of prognostic factors of radiotherapy in children with Ⅲ-Ⅳ neuroblastoma Jun-jun ZHOU, Jie ZHAO, Jing-yan TANG, Ma-wei JIANG, Xiu-mei MA, Yong-rui BAI 2021, 41 (8):  1051-1055.  doi: 10.3969/j.issn.1674-8115.2021.08.009 Abstract ( 739 )   HTML ( 90 )   PDF (991KB) ( 482 )   ·To analyze the factors affecting the survival and prognosis of the children with medium-, high-, and very high-risk neuroblastoma (NB) in stage Ⅲ?Ⅳ after radiation therapy. ·A total of 132 cases of neuroblastoma patients from 2008 to 2018 who received local radiotherapy in the Department of Hematology and Oncology, Shanghai International Children′s Medical Center, Shanghai Jiao Tong University School of Medicine, and the Department of Radiation Oncology, Renji Hospital and Xinhua Hospital were included. Treatment options included chemotherapy, surgery, autologous bone marrow transplantation (ABMT), radiotherapy, and later 13-cis-RA maintenance therapy. SPSS 19.0 software was used for the calculation of event-free survival (EFS) and overall survival (OS) rates, as well as univariate and multivariate survival analysis. ·By the end of the follow-up, the EFS rates for the all patients were 87.41% in 1 year, 55.25% in 2 years, 45.02% in 3 years, and 38.67% in 5 years, and the OS rates were 96.02% in 1 year, 83.54% in 2 years, 72.15% in 3 years, and 57.79% in 5 years, respectively. The medium follow-up time was 40.3 months. Univariate survival analysis suggested that the staging, risk grouping, brain metastasis or not, blood lactic acid dehydrogenase and serum ferritin levels were associated with EFS and OS (P<0.05). The presence or absence of bone marrow infiltration was a related factor of EFS (P=0.007), but not OS-related. Factors such as the child′s gender, whether the child was treated with ABMT, the amplification of MYCN proto-oncogene, and the presence of liver metastasis had no significant correlation with EFS or OS. Multivariate survival concluded that brain metastasis was an independent prognosis factor for EFS and OS. ·The amplification of MYCN gene and ABMT treatment may not significantly affect the radiotherapy efficacy and overall survival of stage Ⅲ?Ⅳ NB children. However, the presence of brain metastasis is an independent prognostic factor related to survival and disease progression after radiotherapy. Figures and Tables | References | Related Articles | Metrics

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Expression and clinical significance of HIF-1α/SHH signaling pathway in neuroblastoma Sheng CHEN, Zhi-long YAN, Ye-ming WU, Min XU, Song GU, Jing MA 2021, 41 (8):  1056-1061.  doi: 10.3969/j.issn.1674-8115.2021.08.010 Abstract ( 739 )   HTML ( 5 )   PDF (2806KB) ( 371 )   ·To investigate the expression and clinical significance of hypoxia inducible factor-1α (HIF-1α)/sonic hedgehog (SHH) signaling pathway in neuroblastoma (NB). ·The expressions of HIF-1α and SHH in paraffin sections of 101 NB cases and 20 ganglioneuroma (GN) cases were analyzed by immunohistochemistry. The differences in the expression levels of these two proteins were compared between NB and GN. And correlations between HIF-1α/SHH expression levels and NB patients′ clinicopathological characteristics or prognosis were analyzed. ·The expression levels of HIF-1α and SHH in NBs were significantly higher than those in GNs (HIF-1α: 26.7% vs 5.0%, P=0.035; SHH: 32.7% vs 10.0%, P=0.041). Compared with low-stage (stage 1 and 2) NBs or NBs with favorable histology, HIF-1α and SHH expression levels were significantly higher in high-stage (stage 3 and 4) NBs or NBs with unfavorable histology (HIF-1α: P=0.002 and 0.012, respectively; SHH: P=0.021 and 0.007, respectively). The expression levels of these two proteins were also higher in NBs with local lymph node metastasis (HIF-1α: P=0.012; SHH: P=0.001). A significant positive correlation was found between HIF-1α and SHH expression levels in NB specimens (r=0.41, P=0.000). The 5-year overall survival (OS) rate and event-free survival (EFS) rate of the HIF-1α high-expression group were significantly lower than those of the HIF-1α low-expression group(OS rate: 40.9% vs 75.7%, P=0.001; EFS rate: 40.9% vs 70.0%, P=0.008). The 5-year OS rate and EFS rate of the SHH high-expression group were also significantly lower than those of the SHH low-expression group (OS rate: 42.9% vs 78.1%, P=0.000; EFS rate: 39.3% vs 73.4%, P=0.001). ·The increased expressions of HIF-1α and SHH in NB are closely related to clinical features of NB, such as stage, local lymph node metastasis and unfavorable histology, and indicate the poor prognosis. Hence, HIF-1α/SHH signal pathway might be a potential biomarker and therapeutic target for NB. Figures and Tables | References | Related Articles | Metrics

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Analgesic effect of multiple spinal nerve block on video-assisted thoracoscopic segmentectomy or lobectomy Zi-han WANG, Jie SHEN, Yan LUO 2021, 41 (8):  1062-1067.  doi: 10.3969/j.issn.1674-8115.2021.08.011 Abstract ( 405 )   HTML ( 4 )   PDF (1752KB) ( 330 )   ·To investigate the analgesic effect and clinical safety of multiple spinal nerve block in incision pain and visceral pain in patients undergoing video-assisted thoracoscopic segmentectomy or lobectomy. ·From May 2020 to October 2020, 121 consecutive general anaesthesia patients scheduled for video-assisted thoracoscopic segmentectomy or lobectomy in Ruijin Hospital, Shanghai Jiao Tong University School of Medicine were allocated to thoracic paravertebral block (TPVB) group (group T, n=60) or multiple spinal nerve block group (group M, n=61). Group T received ultrasound-guided TPVB on the T5 level with 40 mL 0.3% ropivacaine. Group M received ultrasound-guided multiple spinal nerve block including serratus anterior plane block on the 5th intercostal space with 10 mL 0.3% ropivacaine, erector spinae plane block on the T5 level with 10 mL 0.3% ropivacaine, retrolaminar block on the T5 level with 10 mL 0.3% ropivacaine, and retrolaminar block on the T3 level with 10 mL 0.3% ropivacaine. Perioperative hemodynamic characters, postoperative Numeric Rating Scale (NRS) scores of incision pain and visceral pain, and major adverse events were compared between the two groups. ·There was no significant difference in clinical characteristics, perioperative hemodynamic characters, and the use and dose of intraoperative analgesics or vasoactive drugs between the two groups (P>0.05). The NRS scores of incision pain and visceral pain in group M were lower than those in group T during postoperative 24 h (Calm: incision pain, P=0.001; visceral pain, P=0.000. Coughing: incision pain, P=0.000; visceral pain, P=0.000). NRS scores of visceral during pain postoperative 48 h in group M were significantly lower than those of group T (P=0.006) when coughing. Compared with group T, the pressing times of patient-controlled intravenous analgesia pump and the usage of flurbiprofen decreased in group M within 48 h after operation (both P=0.000), and the time of first postoperative pain was delayed (P=0.002). The postoperative activity time was advanced (P=0.000), and the incidence of vomiting decreased within 48 h after operation (P=0.025). ·Multiple spinal nerve block has better analgesic effect on video-assisted thoracoscopic segmentectomy or lobectomy than TPVB, which relieves both incision pain and visceral pain. There is no difference in clinical safety between the multiple spinal nerve block and TPVB. Figures and Tables | References | Related Articles | Metrics

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Preliminary analysis of retinal microvasculature and thickness in constant exotropia adults Li-ying LUO, Min TANG, Xiao-qiong XIANG, Yang FU 2021, 41 (8):  1068-1073.  doi: 10.3969/j.issn.1674-8115.2021.08.012 Abstract ( 806 )   HTML ( 13 )   PDF (3330KB) ( 320 )   ·To investigate the retinal microvascular and thickness abnormalities in adult patients with constant exotropia using OCTA (optical coherence tomography angiography) device, and explore the clinical application value of OCTA in constant exotropic adults. ·A total of 29 adult patients with constant exotropia who visited the Department of Ophthalmology, Shanghai General Hospital, Shanghai Jiao Tong University from January to December 2019 were recruited as the experimental group. Twenty-seven eyes of 27 healthy volunteers who visited the hospital during the same period were included as the control group. All participants underwent a comprehensive ophthalmologic examination at the first visit, which included subjective refraction, intraocular pressure measurement, ocular motility and position test, cover test at distance and near, axial length, and slit-lamp examination. Pearson's chi-squared test was used to compare categorical variants. The vessel and thickness in 3 mm×3 mm macular area were detected by OCTA device by a single experienced technician. The average vessel density of superficial capillary plexus (SCP), deep capillary plexus (DCP), foveal avascular zone (FAZ) area and inner and full retina thickness were automatically calculated by software. The differences in OCTA parameters between the dominant eyes, non-dominant eyes and control eyes were compared by using t-test or Mann-Whitney U test. The OCTA data was corrected for axial length and refractive error using Littmann's formula. A value of P<0.05 was considered statistically significant. ·There was no significant difference in age and gender between the experimental group and the control group. All participants had normal eye movements and normal anterior and posterior segments of the ocular. There were no statistically significant differences in SE, BCVA, intraocular pressure, and axial length of the dominant, non-dominant and normal eyes. In the 3 mm×3 mm macular region, there was no significant difference in the average blood flow density of SCP, DCP and FAZ area in the dominant, non-dominant and control eyes. The fovea and temporal regions of full macular thickness of the non-dominant eyes were (248.61±19.84) μm and (320.44±17.05) μm respectively, significantly (P=0.018, P=0.018) thinner when compared with the dominant eyes [(251.61±19.37) μm, (323.44±15.82) μm]. ·OCTA is an effective tool to quantitatively analyze the blood flow density, FAZ area and macular thickness of patients with constant exotropia. There are no significant differences in the average blood flow density of SCP, DCP and FAZ area in adult patients with constant exotropia; there are specific changes in the macular microstructure of the non-dominant eyes. The fovea and temporal regions of full macular thickness of the non-dominant eyes are significantly decreased. Figures and Tables | References | Related Articles | Metrics

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Correlation between genotype and phenotype of inherited factor Ⅴ deficiency Ke-ke LI, Zhao-lin CHEN, Ying FENG, Yang XIAO 2021, 41 (8):  1074-1080.  doi: 10.3969/j.issn.1674-8115.2021.08.013 Abstract ( 631 )   HTML ( 5 )   PDF (1695KB) ( 445 )   ·To perform clinical phenotype diagnosis,thrombin generation assay (TGA) and F5 analysis in FⅤD patients, evaluate clinical bleeding risk and discuss the correlation between genotype and phenotype. ·Five patients with FⅤD who visited the Department of Hematology of The Second Affiliated Hospital of Guangzhou Medical University from January to November 2020 were selected. PT and APTT were detected by routine coagulation screening. Plasma FⅤ∶C and plasma FⅧ∶C were detected by one-stage coagulation. Plasma FⅤ∶Ag, free and total TFPI was measured by ELISA. The risk of bleeding was graded according to the bleeding score recommended by ISTH.TGA was used to evaluate the risk of clinical bleeding in the patients with FⅤD. All exons and flanking sequences of F5 were analyzed by Sanger sequencing, and the mutation sites were verified by reverse sequencing. AccuCopy multiple gene copy number detection technique was used to detect F5 copy number variation (CNV) in patients with FⅤD. ·All 5 probands belonged to type Ⅰ FⅤD, including 2 cases of severe FⅤD (FⅤ∶C <1%). The levels of free and total TFPI decreased in varying degrees. The lagtime and time to peak of TGA were significantly prolonged, but the levels of peak height and endogenous thrombin potential varied. Mutation analysis revealed 8 kinds of mutations, including missense, nonsense, frameshift mutations and CNV, of which missense mutations accounted for 75% of the total mutations. p.Cys603Ser, p.Leu949Trpfs*, p.Leu1262_Gln1657del were novel mutations. Large fragment deletions in exon 13?14 of proband 2 were detected by AccuCopy (c.3784_4971del, p.Leu1262_Gln1657del). Further analysis of mRNA level showed that the large fragment deletion mutation led to abnormal mRNA splicing, resulting in novel splice site with three kinds of abnormal transcripts (c.3577_4971del, c.3577_4456del, c.3331_4456del). ·The level of FⅤ∶C in patients with severe FⅤD is not related to the severity of bleeding, but TGA and bleeding score can well correlate the bleeding risk. The bleeding severity of FⅤD may be related to the type of F5 mutation. Figures and Tables | References | Related Articles | Metrics

Public health

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Development and analysis of reliability and validity of Operating Room Nurses' Job Stressor Scale Wen-jing QIAN, Ya-qing ZHANG 2021, 41 (8):  1081-1088.  doi: 10.3969/j.issn.1674-8115.2021.08.014 Abstract ( 771 )   HTML ( 17 )   PDF (986KB) ( 807 )   ·To develop a scale for measurement of operating room nurses′ job stressor in China and analyze its reliability and validity. ·According to the literature review, combined with the working characteristics of the nurses in operating rooms, the first draft with 47 items was formulated by two groups of focus interviews among the operating room nursing managers and the operating room nurses. And the items were demonstrated and modified by Delphi expert consultation method to formulate the test version of Operating Room Nurses′ Job Stressor Scale. A test was conducted among the 355 nurses in operating rooms in three tertiary level A hospitals in Shanghai. The items were further screened and the scale was optimized according to the results of item analysis and exploratory factor analysis, and the reliability and the validity of the scale were analyzed. ·Twelve items were deleted and 8 items were modified by the first round of expert consultation. The test version of Operating Room Nurses′ Job Stressor Scale with 35 items was formulated by the second round of expert consultation. Item-level content validity indexes (I-CVI) were all ≥0.78 and scale-level average content validity index (S-CVI/Ave) was 0.91. A total of 355 questionnaires were distributed and the effective recovery rate was 100%. Thirty-five items were retained after item analysis. After 7 times of exploratory factor analyses, 6 items were deleted and 29 items were remained. A total of four factors were extracted ( ??problems in surgical nursing and patient nursing'' ??problems in working environment, workload and time allocation'' ??problems in nursing specialty and development'' and ??problems in management and interpersonal relationship''). The cumulative variance explanation rate of the four factors was 72.162%. The correlation coefficient between each dimension and the total score was higher than that between dimensions. S-CVI/Ave was 0.92. The internal consistency Cronbach′s α of the scale was 0.969, and those of the four dimensions were 0.936, 0.938, 0.872, and 0.943, respectively. The split-half reliability Spearman-Brown coefficient was 0.906. The retest reliability coefficient was 0.946. ·Operating Room Nurses′ Job Stressor Scale has good reliability and validity, which can be used to measure the source and degree of working pressure for the nurses in operating rooms. Figures and Tables | References | Related Articles | Metrics

Review

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Research progress in the development of osteoarthritis mediated by pericellular matrix Wen-cheng HU, Hong-yi ZHU, Jun-qing LIN, Xian-you ZHENG 2021, 41 (8):  1089-1093.  doi: 10.3969/j.issn.1674-8115.2021.08.015 Abstract ( 694 )   HTML ( 9 )   PDF (810KB) ( 374 )   Osteoarthritis (OA) is one of the most common degenerative diseases in clinic. It causes joint pain and dyskinesia, and seriously affects the quality life of the patients′. However, the pathogenesis of OA has not been fully understood. Pericellular matrix (PCM) is a narrow matrix area around chondrocytes. Numerous evidences have revealed the promotion of PCM in the pathogenesis of osteoarthritis. The degradation of PCM components, the release of growth factor and the alteration of mechanical signal transduction accelerate the occurrence and development of OA. This paper summarizes the structure and functions of PCM, and reviews the latest progresses in the involvement of PCM in OA pathogenesis. References | Related Articles | Metrics

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Research progress in epigenetics in methamphetamine use and addiction Ying-jie CHENG, Qian-qian SUN, Min ZHAO 2021, 41 (8):  1094-1098.  doi: 10.3969/j.issn.1674-8115.2021.08.016 Abstract ( 961 )   HTML ( 27 )   PDF (818KB) ( 948 )   Methamphetamine is a central nerve psychostimulant with strong addictive effects and neurotoxicity. However, the mechanism of methamphetamine addiction is still unknown. Epigenetics regulates gene expression without influencing DNA sequence and is a research hotspot in recent years. Increasing researches indicate that epigenetics may participate in methamphetamine-induced brain structure and function alterations. This may provide new insights for exploring the pathogenesis of methamphetamine addiction. This article reviews the advances of epigenetic researches related to methamphetamine use and addiction. References | Related Articles | Metrics

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Biological and angiogenic effects of enamel matrix derivative on periodontal regeneration-related cells Chi-hsiang CHUANG, Jia-chen DONG, Rong SHU 2021, 41 (8):  1099-1102.  doi: 10.3969/j.issn.1674-8115.2021.08.017 Abstract ( 668 )   HTML ( 11 )   PDF (755KB) ( 490 )   Enamel matrix protein has been proved to promote periodontal regeneration, and later developed into enamel matrix derivative (EMD). At present, EMD has been used in clinic for more than 20 years, and a number of clinical studies have shown that it also has a prominent role in periodontal regeneration. Angiogenesis also plays an essential role in tissue regeneration. This process allows the transportation of numerous cytokines to tissue defect areas via newly formed blood vessel, which can then stimulate periodontal regeneration. Several researches have indicated that EMD can stimulate angiogenesis. In this paper, the effects of EMD on periodontal regeneration-related cells and angiogenesis are reviewed. References | Related Articles | Metrics

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Review of immunosuppressive tumor microenvironment of pancreatic cancer Jing-wei LI, Li-wen WANG, Ling-xi JIANG, Qian ZHAN, Hao CHEN, Bai-yong SHEN 2021, 41 (8):  1103-1108.  doi: 10.3969/j.issn.1674-8115.2021.08.018 Abstract ( 2098 )   HTML ( 142 )   PDF (908KB) ( 1483 )   Pancreatic cancer is a highly malignant tumor. The difficulty of early diagnosis and scarcity of effective clinical treatment strategies lead to poor prognosis. Tumor microenvironment (TME) of pancreatic cancer is composed of tumor cells, immune cells, stromal cells, extracellular matrix and soluble factors. TME plays an important role in the development, progression, invasion and metastasis of tumors. The pancreatic cancer microenvironment has significant immune cell infiltration, which is highly immunosuppressive. On the one hand, tumor cells edit the immune system so that cancer cells cannot be recognized by the immune system; on the other hand, they can recruit and activate various immunosuppressive cells such as pancreatic stellate cells, myeloid-derived inhibitory cells, tumor-associated macrophages, regulatory T cells and so on. These immunosuppressive cells can secrete immunosuppressive molecules, affect the function of anti-tumor immune cells, inhibit the host′s anti-tumor immune response, lead to tumor immune escape, and promote tumor development and metastasis. In this review, the mechanisms and effects of these immunosuppressive components are discussed and the updated results of immunotherapy on pancreatic cancer are studied, which may provide novel insights on TME and immunotherapy of pancreatic cancer. References | Related Articles | Metrics

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Application of OCT and OCTA to chronic ocular ischemic diseases caused by carotid stenosis Gong CHEN, Xi SHEN 2021, 41 (8):  1109-1113.  doi: 10.3969/j.issn.1674-8115.2021.08.019 Abstract ( 692 )   HTML ( 106 )   PDF (932KB) ( 451 )   Carotid stenosis is one of the major causes of chronic ocular ischemic diseases, which are often manifested as venous stasis retinopathy and ocular ischemic syndrome, and the latter can lead to neovascular glaucoma. At present, fundus fluorescein angiography, indocyanine green angiography, color Doppler flow imaging and carotid artery imaging help to establish the diagnosis. Recently, optical coherence tomography (OCT) and optical coherence tomography angiography (OCTA) have been gradually applied to make diagnosis and follow-up in chronic ocular ischemic diseases caused by carotid stenosis as novel, non-invasive, rapid and high-resolution techniques. This review summarizes recent applications of OCT and OCTA to chronic ocular ischemic diseases caused by carotid stenosis. References | Related Articles | Metrics

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Research progress in screening non-small cell lung cancer patients who will benefit from immunotherapy Xu-xin-yi LING, Yao ZHANG, Hua ZHONG 2021, 41 (8):  1114-1119.  doi: 10.3969/j.issn.1674-8115.2021.08.020 Abstract ( 654 )   HTML ( 11 )   PDF (916KB) ( 427 )   The application of immune checkpoint inhibitors (ICIs) has brought the treatment of non-small cell lung cancer (NSCLC) into a brand-new age. However, not all patients with NSCLC can benefit from ICIs, and there is currently no good standard for screening the beneficiaries, which provides difficulties for ICIs to bring greater benefits to patients. Therefore, it is quite important to explore suitable biomarkers that can predict the level of patients′ response to ICIs. Due to the complexity of the mechanism of the action of ICIs, many links involved in the tumor immune response, including tumor-related cells, molecules or genetic characteristics, may affect the patients' response to ICIs, and may also have the ability to predict the level of NSCLC patients' response to ICIs at the same time. Currently many studies are exploring the clinical significance and limitations of existing biomarkers, struggling to explore potential emerging biomarkers and combine multiple biomarkers for a more reliable predicting model, in order to bring precise treatment to patients with NSCLC. This article is a review based on the latest research results. It summarizes the current application and research progress of biomarkers to screen NSCLC candidates who will benefit from immunotherapy. References | Related Articles | Metrics

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Research progress of DNA methylation in gestational diabetes mellitus Dan WU, Li-ping GE 2021, 41 (8):  1120-1124.  doi: 10.3969/j.issn.1674-8115.2021.08.021 Abstract ( 735 )   HTML ( 16 )   PDF (824KB) ( 742 )   Gestational diabetes mellitus refers to patients with normal glucose metabolism before pregnancy and getting abnormal glucose tolerance during pregnancy. It is characterized by decreased insulin sensitivity of adipose and muscle tissues and abnormal blood glucose levels in the body. This disease can seriously affect the maternal and fetal health, and the risk of type 2 diabetes mellitus and other metabolic diseases in the postpartum period and the offsprings significantly increased. It has been found that epigenetic modifications such as DNA methylation can be involved in the pathogenesis of gestational diabetes mellitus and the occurrence of maternal and fetal complications. The differences in gene methylation levels appear in the various clinical specimens of patients with gestational diabetes mellitus, such as placentas, umbilical cord blood, peripheral blood and adipose tissues. It can also affect all the lives of the offsprings through “fetal programming”. In this paper, the research progress of DNA methylation in gestational diabetes mellitus is reviewed. Figures and Tables | References | Related Articles | Metrics

Brief original article

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Effects of different continuous infusion rates of cisatracurium on muscle relaxation and recovery of muscle relaxation during upper abdominal laparotomy Kai HE, Yan LUO, Chun-mei XU, Wei-fang JIN 2021, 41 (8):  1125-1128.  doi: 10.3969/j.issn.1674-8115.2021.08.022 Abstract ( 725 )   HTML ( 90 )   PDF (866KB) ( 584 )   ·To investigate the effects of different continuous infusion rates of cisatracurium on muscle relaxation and recovery of muscle relaxation in upper abdominal laparotomy. ·From January 2019 to January 2020, 60 patients scheduled for upper abdominal laparotomy in Shanghai Civil Aviation Hospital/Gubei Branch of Ruijin Hospital were randomly divided into low dosage [1.0 μg/(kg·min)] group (L group), middle dosage [1.5 μg/(kg·min)] group (M group) and high dosage [2.0 μg/(kg· min)] group (H group), according to the speed of intraoperative pump injection of cisatracurium, with 20 cases in each group. The amount of cisatracurium during the induction period, the cumulative pumping dosage during the operation, intraoperative muscle relaxation satisfaction, the time from drug withdrawal to the train of four stimulation (TOF) ratio recovery to 0.25, 0.75, 0.80 and 0.90 (TOFr 0.25, TOFr 0.75, TOFr 0.80, TOFr 0.90) and extubation time were recorded and compared among the three groups. ·There was no significant difference in the dosage of cisatracurium during induction among the three groups (P=0.662). The cumulative pump the dosage in the L group was lower than those in the M and H groups (P=0.025). There was no significant difference in intraoperative muscle relaxation satisfaction among the three groups (P=1.000). TOFr 0.25, TOFr 0.75, TOFr 0.80, TOFr 0.90 and extubation time in the L group were significantly shorter than those in the M and H groups (P=0.000), but there was no significant difference between M group and H group (P>0.05). ·Continuous infusion of cisatracurium at the rate of 1.0 μg/(kg·min) during upper abdominal laparotomy can provide a good state of muscle relaxation. The increase of pump dosage can not significantly improve the effect of muscle relaxation, but can prolong the recovery time of muscle relaxation and extubation time. Figures and Tables | References | Related Articles | Metrics

Case report

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A case report of Peutz-Jeghers syndrome with jejunal intussusception Ming-wang LUO, Zhao-qi ZHANG, Yuan WEI, Wen-di YAN, Dong-wang YAN 2021, 41 (8):  1129-1132.  doi: 10.3969/j.issn.1674-8115.2021.08.023 Abstract ( 619 )   HTML ( 9 )   PDF (2472KB) ( 390 )   A female patient, 16-year-old, presented ??recurrent lower abdominal pain for one year, recurrence and aggravation for 10 days''. Physical examination revealed that the patient's lips had dot-like dark spots, 1?4 mm in diameter, with clear boundaries and without fusion and fading under pressure, and there was tenderness in the upper left abdomen. The laboratory and imaging examinations were further completed and the patient was admitted to hospital with ??1. intussusception; 2. multiple polyps in the intestine; 3. likely Peutz-Jeghers syndrome (PJS)''. After admission, laparotomy was performed for duodenal and jejunal polyp resection, and the postoperative diagnosis was PJS. PJS is a clinically rare disease with three typical characteristics: skin and mucous membrane pigmentation, multiple polyps in the gastrointestinal tract, and family genetic tendency. The clinical data of this case are introduced to improve the understanding of PJS, provide reference for the following diagnosis and treatment of similar patients, and reduce the occurrence of emergency surgery and short bowel syndrome. Figures and Tables | References | Related Articles | Metrics