Table of Content

05 July 2021, Volume 41 Issue 7

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Original Articles

Ghrelin improves endothelial function and plaque stability after myocardial infarction in ApoE^-/- mice

WANG Li, PANG Jun, CHEN Qing-wei, LI Gui-qiong, KE Da-zhi, LI Xing-sheng

2021, 41(7):  935-940. 

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Objective To investigate the effects and mechanisms of ghrelin on plaque stability and endothelial funtion in ApoE^-/- mice after myocardial infarction. Methods The ApoE^-/- mice were divided into control group, double model group and ghrelin-treated group. All ApoE^-/- mice were fed with a high-fat diet for 12 weeks to induce atherosclerotic vulnerable plaques. The atherosclerotic vulnerable plaque model was the control group.At the 8th week of this study, the double model group and the ghrelin group were subjected to acute myocardial infarction model(AMI). After modeling of AMI, the ghrelin group was administered with ghrelin (100 μg/kg,bid) until the end of the 12th week. Body weight and blood lipids were detected. Left ventricular ejection fraction (LVEF) and left ventricular fractional shortening (LVFS) were recorded by echocardiography; NO level was measured by Griess method; the percentage of aortic sinus plaque area was evaluated by HE microscopy. The plaque content of lipid and collagen was observed by Oil Red 0 and Sirius red staining; The distribution of macrophages and smooth muscle cells in plaques were determined by RAM-11 and α-actin immunohistochemical staining and microscopy; And then the vulnerability index was calculated; Myocardial infarct size was measured by Masson staining and microscopy; The vascular endothelial growth factor (VEGF) was detected by quantitative real-time PCR and Western blot. Results Compared with the control group, the level of LVEF and LVFS and NO₂^-/NO₃^- concentration in the double model group were all decreased(P＜0.05), and the expression of VEGF was increased(P＜0.05), significantly. However, the ghrelin group had significantly reduced plasma TG level and also the myocardial infarct size(P＜0.05), LVEF, LVFS, NO₂^-/NO₃^- level, the expression of VEGF, and the plaque content of collagen and smooth muscle cells were all increased(P＜0.05), and reduced the percentage of aortic sinus plaque area, the distribution of lipid and macrophages(P＜0.05), and the vulnerability index as compared with the double model group(P＜0.05). Conclusions 1)Ghrelin may improve the endothelial function and improve heart function in ApoE^-/- mice after myocardial infarction. 2)Ghrelin may stabilize vulnerable plaque and reduce the occurrence of reinfarction.

Bletilla striata polysaccharide up-regulates the expression of tight junction protein occludin in intestinal mucosa of mice with ulcerative colitis

LI Yu-xin, YU Xue-yan, HUANG Xue

2021, 41(7):  941-945. 

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Objective To investigate the effect of Bletila striata polysaccharide(BSP) on ulcerative colitis (UC) mice induced by dextran sodium sulfate (DSS) and its effect on tight junction occludin protein in intestinal mucosa. Methods Mice were randomly divided into control group, DSS model group (freely drinking 2.5% DSS solution for 7 days), salazosulfopyridine(300 mg/kg) positive control group, BSP low-dose (125 mg/kg) and high-dose groups (250 mg/kg). From the beginning of modeling, mice were given intragastric administration once a day for consecutive 7 days. The change of body weight, disease activity index (DAI) score, histopathology score and colon length were recorded to evaluate the inflammatory reaction. Expression of occludin protein in mice colon tissue was detected by Western blot. Results Compared with control group, the body weight of the model group mice was significantly reduced, the colon was shortened, the DAI and histopathology scores were significantly increased(P＜0.01), and the expression of occludin protein in colon tissue was significantly reduced (P＜0.01). Compared with DSS model group, BSP significantly alleviated body weight loss and colon shortening, and decrease DAI and histological scores, and up-regulated the expression of occludin protein in colon tissue of UC mice. Conclusions BSP alleviates inflammatory reactions of colonic mucosa in UC mice. The mechanism may be the improvement of intestinal epithelial barrier function mediated by increased expression of colonic mucosa occludin protein.

Knockdown of Ube2w increases the susceptibility to dextran sodium sulfate-induced ulcerative colitis in mice

WANG Shao-xin, CUI Li-hong, LI Xiao-wei, LIU Xin-yao, LUO Zhe, YAN Zhi-hui

2021, 41(7):  946-950. 

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Objective To investigate the effect of UBE2W on dextran sodium sulfate (DSS)-induced ulcerative colitis in mice. Methods Different interference sequences of UBE2W siRNA(1#, 2# and 3#) were transiently transfected into mouse fibroblast cell line L929.The expression of UBE2W protein was detected by Western blot. The siRNA with the most significant inhibitory effect was selected for transfection in vivo. Ten C57BL / 6 male mice were randomly injected with siRNA (siUbe2w/si-control) via caudal vein and fed with 2.5% DSS to construct two groups of DSS ulcerative colitis models, Ube2w knockdown group and control group with 5 mice in each group. Western blot and RT-qPCR were used to detect the expression of UBE2W in colon tissue of mice. The weights of mice in the two groups were observed, and the severity of colitis was compared by histological score. Results Ube2w siRNA 2# inhibited UBE2W protein expression most significantly in L929 cells. UBE2W mRNA and protein expression in colon tissue of Ube2w knockdown group were lower than those in control group, and weight loss in Ube2w knockdown group was significantly more obvious than that in control group, and the histological score of colitis in knockdown group was 8.60±0.24, which was higher than that in control group (5.20±0.49)(P＜0.05). Conclusions UBE2W is involved in the regulation of inflammatory response and Ube2w knockdown increases the susceptibility to DSS induced ulcerative colitis in mice.

Sodium hydroxide enhances the efficacy of rapamycin to suppress proliferation of Tsc2-null mouse embryonic fibroblasts

LI Kai, WANG Ya-nan

2021, 41(7):  951-956. 

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Objective To explore the effect of sodium hydroxide(NaOH) and rapamycin(Rapa) alone or their combination in treating Tsc2-null(Tsc2^-/-) mouse embryonic fibroblasts. Methods The wild type and Tsc2^-/- cells were treated with NaOH and/or Rapa and divided into 4 groups: control, NaOH group, Rapa group, and combination (Combo) group. Cell proliferation was measured by MTT assay and colony formation. Flow cytometry was used to detect the potential influence on cell cycle. The protein expression of S6, p-S6, p-4EBP1 was detected by Western blot. Results Compared to control group, Tsc2^-/- cells in NaOH and Rapa groups both showed a reduced proliferation ability and colony counting (P＜0.05); additionally, only in Rapa group, the percentage of Tsc2^-/- cells in G1 phase increased and the protein level of p-S6 and p-4EBP1 all decreased (P＜0.05). Compared with NaOH and Rapa groups, Tsc2^-/- cells in Combo group had a further decreased in proliferation ability and colony number and an increased G1 phase percentage (P＜0.05). But the level of p-S6 and p-4EBP1 of Tsc2^-/- cells in Combo group was significantly lower than those in NaOH group (P＜0.05). Conclusions NaoH(15 μmol/L) enhances the suppressing poliferation efficacy of Rapa in Tsc2^-/- cells.

Carboxyamidotriazole-orotate inhibits the proliferation and mitochondrial energy metabolism of mouse glioma cell line GL261

HUANG Yu-qing, GAO Hong-ting, YANG Li-xing, CHEN Qiu-xia, WANG Yu-cheng, JU Rui, GUO Lei

2021, 41(7):  957-962. 

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Objective To investigate the effects of carboxyamidotriazole-orotate (CTO) on the proliferation and apoptosis of mouse glioma cell line GL261. Methods The proliferation of GL261 cells was detected by live cell counting. Cell cycle of GL261 cells was detected by PI staining and flow cytometry. Apoptosis of GL261 cells was detected by annexinⅤ/PI double staining and flow cytometry; DCFH-DA staining and flow cytometry were used to detect the content of intracellular reactive oxygen species (ROS) of GL261 cells. The oxygen consumption rate (OCR) of GL261 cells was measured in Seahorse bioenergy assay. The ATP content in GL261 cells was detected by ATP detection kit. Results Compared to the control group, the proliferation of GL261 cells in CTO treatment group was significantly inhibited in a time- and dose-dependent way, the proportion of GL261 cells in S phase was significantly increased (P＜0.05) in 20 μmol/L CTO group. Compared with the control group, more apoptosis occurred in GL261 cells of CTO group, and ROS content in GL261 cells of 20 μmol/L CTO group was also significantly increased (P＜0.01). Compared with the control group, the contents of OCR and ATP in GL261 cells was signifi- cantly reduced in the CTO group(P＜0.01). Conclusions CTO inhibits proliferation of GL261 cells and promotes their apoptosis by increased production of ROS. In addition, CTO inhibits the mitochondrial respiration of GL261 cells and the production of ATP in mitochondria, thus inhibits growth of glioma cells.

Knocking out Fcgr1 of dorsal root ganglion attenuates the activation of NF-κB/NLRP3 pathway in a rat model of rheumatoid arthritis

LIU Fan, SU Si, WANG Tao, YUAN Bo, MA Chao

2021, 41(7):  963-969. 

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Objective To explore the mechanism of peripheral neurons FcγRⅠ regulating pain caused by rheumatoid arthritis. Methods Rheumatoid arthritis models were established in 14 wild-type SD rats and 7 Fcgr1 conditional knockout rats. Wild-type rat control group (control group), rheumatoid arthritis group of wild-type rat (RA group) and rheumatoid arthritis group of Fcgr1 conditional knockout rat (RA+CKO group) were established. Pain behavior was used to detect the changes of mechanical pain threshold and thermal pain threshold 3 and 1 days before modeling and 3, 5, 7 and 9 days after modeling. The expression of Fcgr1 mRNA in dorsal root ganglion (DRG) of Fcgr1 conditional knockout rat was detected by fluorescence in situ hybridization. Immunofluorescence staining was used to detect the changes of IL-1β and IL-18 expression in the DRG and activation of glial cells in the spinal dorsal horn (SDH). Results Compared with the control group, the threshold of mechanical and thermal pain in RA group and (RA+CKO) group were decreased (P＜0.05), the mechanical and thermal pain thresholds of the (RA+CKO) group were significantly higher than of RA group(P＜0.05). Compared with the RA group, the expression of pNF-κB, NLRP3, IL-1β and IL-18 and the protein level of GFAP and Iba1 in the SDH of (RA+CKO) rats were significantly decreased (P＜0.05). Conclusions FcγRⅠ of DRG neurons may promote the synthesis and release of inflammatory cytokines IL-1β and IL-18 through NF-κB/NLRP3 pathway, and cause glial cells activation in the SDH, which may play a role in the mechanism causing pain of rheumatoid arthritis.

Effects of Colla corii asini on lung function and pathological injury in rats with chronic obstructive pulmonary disease

Nazhakaiti·AINIWAER, HU Guang, ZHANG Tian-tian, SONG Mei-yue, ZHAO Hong-mei, JIN Hong-tao, WANG Jing

2021, 41(7):  970-974. 

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Objective To explore the effects of Colla corii asini (CCA) (common name E’jiao) on lung function and pathological injury of the lung tissue in rats with chronic obstructive pulmonary disease (COPD). Methods Twenty-five rats were randomly divided into the control group, COPD model group and treatment group with low, medium and high-doses of CCA (1, 2, 4 g/kg), with 5 rats in each group. The COPD model group and treatment groups were modeled by cigarette smoke exposure with a concentration of (1 100±10)mg/m³ once a day, 90 minutes each time, for 48 consecutive weeks. After the end of the next day, the CCA groups were given by daily oral gavage treatment for 28 days. Lung function was detected by animal lung function instrument and the pathological changes of the lung tissue were assessed by HE staining and microscopy. Results Compared with the control group, TV, EV, Te, RT, EEP and Penh were significantly increased in the COPD model group(P＜0.05), while PEF, PIF, MV, EF₅₀ and f(B) were significantly decreased (P＜0.05). Inflammation and pathological changes of emphysema were observed, and lung tissue lesions were aggravated. Compared with the COPD model group, Te, RT and Penh were significantly reduced in the high-dose CCA group (P＜0.05), while MV, PIF and EF₅₀ were significantly higher (P＜0.05). Less infiltration of inflammatory cells, less alveolar expansion and fewer pulmonary bullae were observed in high-dose CCA-treated rats. The degree of emphysema was reduced, and pathological damage to lung tissue was clearly alleviated. Conclusions CCA improves lung function and alleviates the inflammatory response of the lung tissue in COPD rats.

Isolation and identification of tetraploid trophoblast stem cells

WANG Hai-yu, XIAO Yi, HUANG Yue

2021, 41(7):  975-981. 

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Objective To establish mice tetraploid trophoblast stem cell(TTSCs) line which could be cultured in vitro, providing a new cell model for the research of placenta development. Methods Tetraploid embryos were produced by electrofusion and cultured in vitro, clones were picked up and passaged to establish cell lines. Metaphase chromosome counting was used to detect the chromosome number. RT-qPCR, Western blot and immunofluorescence were used to detect the expression of marker genes. Microinjection was used to detect the development potential in vivo. Results TTSCs that can develop in vivo and be passaged were established. The expression of differentiation genes was distinct between TTSCs diploid TSCs under differentiation condition. Conclusions Passable cell line TTSCs could be established in vitro, but the mechanisms of self-renew and differentiation were different between TTSCs and diploid TSCs.

Recognition of tumor-targeting peptide P7 related to the expression level of Hsp90

KONG De-xin, HUANG Wei, YANG Nan, LIU Yan-yong

2021, 41(7):  982-987. 

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Objective To analyze the correlation between the expression of heat shock protein 90 (Hsp90) and the recognition of LPLTPLP (P7) on different tumor tissues and para-carcinoma tissues. Methods The expression of HSP90 and recognition effects of FITC-P7 in 48 cases of patients with 16 types of organ tumors and matched or mismatched marginal or adjacent tissues were analyzed by immunohistochemistry and immunofluorescence, respectively. Results Hsp90 was highly expressed in tumor tissues than para-carcinoma tissues. FITC-P7 showed a preferred recognition effect on tumor tissues in proportion to Hsp90 expression levels as determined by immunohistochemistry. There was moderate correlation in the two groups of data (r=0.5147,P＜0.01). Conclusions The recognition of tumor tissues by P7 is positively related to the expression level of Hsp90, suggesting the targeting of P7 at Hsp90.

IDH1 regulates the expression of interferon pathway-related gene IFIT3

TANG Wan-jun, PENG Xiao-zhong, HAN Wei

2021, 41(7):  988-994. 

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Objective To explore the role of IDH1 in the expression of RNA binding proteins (RBPs) in glioma cell line LN229. Methods Lentivirus was used to construct IDH1 knockdown and control cells, and the expression of IDH1 was validated by RT-qPCR (real-time quantitative PCR) and Western blot. Then the total RNA of cells from each group was extracted for RNA sequencing (RNA-seq); the differentially expressed genes (DEGs) were identified by limma package and enriched pathways were analyzed; the differentially expressed RBPs were selected according to the RBP database and protein-protein interaction network was constructed by string database; finally, the expression level of candidate RBPs was tested in IDH1 knockdown, over-expressed and mutant cells(R132H) respectively. Results A total of 259 DEGs were detected in IDH1 knockdown LN229 cells, including 146 upregulated and 113 downregulated genes. These DEGs were significantly enriched in interferon related pathways and included 14 RBP-encoding genes, 9 of which were significantly enriched in interferon related pathways and showed close interaction; compared with the control group, IDH1 knockdown in LN229 significantly upregulated the expression of all the nine RBPs (P＜0.001), and IDH1 overexpression only downregulated IFIT3 (P＜0.01) involving in interferon pathway, while IDH1 mutation had no significant effect on IFIT3. Conclusions IDH1 regulates expression of interferon pathway related gene IFIT3 in LN229 cells.

miR-193a-3p inhibits high glucose-induced apoptosis of human retinal vascular endothelial cells

QIN Jian-min, ZHANG Lai-xiang, WEI Zhen-ying

2021, 41(7):  995-1000. 

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Objective To investigate the effect of miR-193a-3p on apoptosis induced by high glucose in human retinal vascular endothelial cells (HRECs) and its molecular mechanism. Methods HRECs cells were cultured in vitro to establish a high glucose (30 mmol/L glucose concentration) cell model. HRECs were divided into control group, model group, anti-miR-NC group, anti-miR-193a-3p group, model+miR-NC group, model+miR-193a-3p group, and model +LY294002 group. RT-qPCR was used to detect the expression of miR-193a-3p; Western blot was used to detect the expression levels of cleaved-caspase-3 (c-caspase-3), B cell lymphoma/leukemia-2(Bcl-2), phosphorylated phosphoinositide 3 kinase (p-PI3K) and phosphorylated protein kinase B (p-AKT); flow cytometry was used to detect cell apoptosis. Results Compared with the control group, the expressions of miR-193a-3p and Bcl-2 in the model group was significantly decreased, whereas apoptosis rate, and the expres- sion of c-caspase-3, P-PI3K and P-Akt was significantly increased (P＜0.05). Low expression of miR-193a-3p promotes c-caspase-3, p-PI3K and p-AKT expression, inhibited Bcl-2 expression, and promoted cell apoptosis (P＜0.05). High expression of miR-193a-3p can significantly reduce the effect of high glucose treatment on the apoptosis, c-caspase-3 and Bcl-2 expression of HRECs (P＜0.05). Inhibition of PI3K/AKT signaling pathway reversed the effect of high glucose treatment the apoptosis, c-caspase-3 and Bcl-2 expression of HRECs (P＜0.05). Conclusions miR-193a-3p could inhibit high glucose-induced apoptosis of HRECs by regulating PI3K/AKT signaling pathway.

LINC00607 affects the proliferation and apoptosis of A549/DDP cells through inhibiting miR-372

YANG Wen-chuan, HAN Dong, YANG Fan

2021, 41(7):  1001-1006. 

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Objective To explore the effect of LINC00607 on proliferation and apoptosis of lung cancer cisplatin-resistant cell line(A549/DDP) and its possible molecular mechanism. Methods The expressions of LINC00607 and miR-372 in lung cancer tissues, adjacent tissues, A549, A549/DDP cells were detected by real-time fluorescence quantitative PCR(RT-qOCR). A549/DDP cells were divided into pcDNA3.1 group, pcDNA3.1-LINC00607 group, (pcDNA3.1-LINC00607 + miR-NC)group, (pcDNA3.1-LINC00607 + miR-372)group. Methyl thiazolyl tetrazolium (MTT) assacy and colony formation experiments were used to test cell proliferation; flow cytometry was used to measure apoptosis; and Western blot was used to detect the expression levels of cleared-caspase-3 and Ki67; Dual luciferase reporter gene and RT-qPCR experiments confirmed the targeted regulation effect of LINC00607 on miR-372. Results Compared with adjacent tissues, the expression of LINC00607 in lung cancer tissues was significantly reduced, while the expression of miR-372 was significantly increased (P＜0.05). Compared with A549 cells, the expression of LINC00607 in A549/DDP cells was significantly reduced, while the expression of miR-372 was significantly increased (P＜0.05). Compared with pcDNA3.1 group, A549/DDP cell survival rate, clone formation numbers, Ki67 protein expression in pcDNA3.1-LINC00607 group were significantly decreased, and apoptosis rate and cleared-caspase-3 protein expression were significantly increased (P＜0.05). Compared with the (pcDNA3.1-LINC00607 + miR-NC)group, the A549/DDP cell survival rate, clone formation, Ki67 protein expression in the (pcDNA3.1-LINC00607 + miR-372)group were significantly increased, and the apoptosis rate, cleared-caspase-3 protein expression were significantly reduced (P＜0.05). LINC00607 targeted at and negatively regulated miR-372 expression. Conclusions LINC00607 inhibits proliferation and induces apoptosis of A549/DDP cells by down-regulating miR-372.

RNAi-reduced bladder TGF-β1 expression improves bladder function and fibrosis in rats with spinal cord injury

JIA Chun-song, ZHAO Yan-po, SHANG Zhen-hua, XING Tian-ying, OU Tong-wen

2021, 41(7):  1007-1012. 

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Objective To explore the effect of reducing bladder TGF-β1 expression with RNA interference(RNAi) on neurogenic bladder(NB). Methods Forty-eight female rats were evenly divided into sham, T10 transaction, LV-vector and LV-shTGF-β1 groups. The sham group underwent sham operation of T10 spinal cord transection, and the other three groups underwent spinal cord transection. The bladder wall was injected immediately after the operation. The first two groups were injected with 0.9% NaCl, the LV-vector group was the control lentivirus, and the LV-shTGF-β1 group was TGF-β1 RNAi lentivirus. On the 28th day after operation, cystometry and bladder weighing were performed, the bladder tissue was stained with HE and Masson, and the expression of detrusor TGF-β1 was detected by immunohistochemistry, RT-qPCR and Western blot. Results In the three groups with spinal cord transection compared with the sham group, the frequency and amplitude of detrusor over activity were significantly increased (P＜0.01), the bladder capacity was significantly reduced (P＜0.05), and the bladder weight, the detrusor interstitial percentage and TGF-β1 expression were significantly increased (P＜0.05). The T10 transection group had no significant difference compared with the LV-vector group. In the LV-shTGF-β1 group compared with the T10 transection group, the frequency and amplitude of detrusor over activity were significantly reduced (P＜0.01), the bladder capacity was significantly increased (P＜0.01), and the bladder weight, the detrusor interstitial percentage and TGF- β1 expression were significantly reduced (P＜0.05). Conclusions Reducing bladder TGF-β1 expression with RNAi can improve detrusor over activity and bladder fibrosis in rats with spinal cord injury.

Down-regulation of perilipin 2 inhibits proliferation of human gastric cancer cell line NCI-N87

LIU Lian-ling, LI Yong

2021, 41(7):  1013-1017. 

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Objective To investigate the expression of perilipin 2 in gastric cancer and its effect on the proliferation, invasion and migration of cancer cells. Methods Human normal gastric mucosa cell line GES-1 and human gastric cancer cell line NCI-N87 were selected. NCI-N87 cells were divided into three groups. control group received no treatment, NC-shRNA group was transfected with 10 μg NC-shRNA recombinant lentivirus plasmid, and perilipin2-shRNA group was transfected with 10 μg perilipin2-shRNA recombinant lentivirus plasmid. The proliferation rate, invasion ability and migration ability of NCI-N87 cells in each group were detected by CCK8 method, Transwell chamber and scratch test, respectively. Western blot was used to detect the expression levels of perilipin2, p62,kelch-like ECH-associated protein 1(keap1), nuclear factor E2-related factor-2(Nrf2) pathway related proteins in NCI-N87 cells in each group. Results The expression level of perilipn2 protein in NCI-N87 cells was significantly higher than that in GES-1 cells(P＜0.05). After treatment for 6,12,24,36 and 48 h, proliferation rate of NCI-N87 cells in perilipin2-shRNA group was lower than that in control and NC-shRNA group(P＜0.05). The number of NCI-N87 cells crossing membrane in perilipin2-shRNA group was less than that in control and NC-shRNA group(P＜0.05). The scratch space in perilipin2-shRNA group was wider than that in control and NC-shRNA group(P＜0.05). The protein expression of perilipn2, p62, keap1 and Nrf2 in NCI-N87 cells of perilipin2-shRNA group was lower than that of control and NC-shRNA group (P＜0.05).Conclusions Perilipin2 is highly expressed in gastric cancer cells, and knockdown of perilipin2 expression can inhibit the proliferation, invasion and migration of gastric cancer cells, and its mechanism may be related to the obstruction of the p62-keap1-Nrf2 pathway.

Anlotinib combined with chloroquine induces apoptosis of human non-small cell lung cancer cell line H1299

LIU Xi-bang, ZHU Lin-zhi, JIAO De-min, TANG Xia-li, CHEN Jun, HU Xu-gang, CHEN Qing-yong

2021, 41(7):  1018-1023. 

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Objective To investigate the effects of anlotinib and chloroquine (CQ) on viability, invasion and apoptosis of H1299 cells. Methods After treatment of H1299 cells with anlotinib or/and CQ, the cell viability was measured by MTT assay; Wound-healing and Transwell chamber assay were used to evaluate cell migration and invasion abilities; Hoechst 33342 staining was adopted to test the changes of apoptosis; The protein levels of autophagy marker LC3-Ⅱ/LC3-Ⅰ and autophagic substrate p62 were detected by Western blot. Results Anlotinib combined with CQ showed more obviously synergistic inhibitory effect on the viability, invasion and migration in H1299 cells than anlotinib alone (P＜0.05). As compared to the untreated control cells, anlotinib alone promoted the cell apoptosis. Their combination induced an even higher apoptosis in H1299 cells. With the increase of concentration, anlotinib significantly up-regulated autophagy marker protein LC3-Ⅱ/LC3-Ⅰ and down-regulated autophagic substrate protein p62.The expression of LC3-Ⅱ/LC3-Ⅰ and p62 was significantly up-regulated by CQ. Compared with the CQ group alone, LC3-Ⅱ/LC3-Ⅰ and p62 were significantly up-regulated and down-regulated in the 20 μmol/L anlotinib combined with CQ group. Conclusions Combination of anlotinib and CQ remarkably inhibits the viability, migration and invasion and induces the apoptosis of H1299 cells. The potential mechanism for antitumor might be due to autophagy's action.

Up-regulation expression of protein kinase D2 reduces lung ischemia-reperfusion injury in mice

LIU Hao-ming, HUA Mao, ZHANG Li, ZHU Hai-hong

2021, 41(7):  1024-1029. 

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Objective To observe the effect of up-regulation of protein kinase D2 (PKD2) expression on lung ischemia-reperfusion injury (LI/RI) in mice. Methods 20 C57BL/6J wild-type (WT) mice and 40 PKD2 over-expression (PKD2^+/+) mice were randomly divided into sham operation group (WS and PS groups), LI/RI group (WIR and PIR groups, the left hilum was first clamped for 1 h, then reperfused for 2 h) and the hypoxia inducible factor-1α (HIF-1α) inhibitor (2-methoxyestradiol, 2-ME) group (WIR+2ME) and (PIR+2ME) groups, 20 mg/(kg·d) 2ME was injected intraperitoneally 2 days before modeling). After 2 hours of reperfusion, the wet/dry weight ratio(W/D) of lung tissues was detected, the pathological changes of lung tissues were observed by HE staining, and the content of inflammatory factors TNF-α, IL-1β and IL-6 in lung tissues were detected by ELISA. RT-qPCR and Western blot were used to detect the expression of (HIF-1α) and vascular endothelial growth factor A(VEGFA) mRNA and protein in lung tissues. Results Compared with WS group, the W/D value of the WIR group increased, the lung tissues were damaged, the content of inflammatory factors increased, and the expression of HIF-1α and VEGFA mRNA and protein increased (P＜0.05). Compared with the WIR group, the PIR group W/D value decreased, lung tissue damage was reduced, inflammatory factor content decreased, HIF-1α and VEGFA mRNA and protein expression increased (P＜0.05). Compared with WIR and PIR groups, W/D values of (WIR+2ME) and (PIR+2ME) groups increased, lung tissue damage worsened, inflammatory factor content increased, HIF-1α and VEGFA mRNA and protein expression significantly decreased (P＜0.05). Conclusions Up-regulation of PKD2 expression can improve LI/RI, and its mechanism may be related to increasing the expression of HIF-1α/VEGFA signaling pathway.

Comparison of different preoperative decompression of biliary tract methods for patients with periampullary carcinoma complicated with obstructive jaundice

CAO Hong-tao, ZHAO Bang-bo, ZHOU Xing-tong, WANG Yuan-yang, LI Tian-hao, QIN Cheng, WANG Wei-bin

2021, 41(7):  1030-1034. 

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Objective To investigate the efficacy of different methods in decompression of biliary tract(jaundice reduction) for patients with periampullary carcinoma combined with obstructive jaundice. Methods The clinical data of 128 patients with malignant jaundice treated by preoperative biliary decompression before pancreaticoduodenectomy were retrospectively analyzed in Peking Union Medical College Hospital from January 2015 to September 2020. Patients were divided into endoscopic retrograde cholangiopancreatography (ERCP) with stent placement group including 80 cases and percutaneous transhepatic cholangial drainage (PTCD) group including 48 cases according to different methods of jaundice reduction. Serological indicators including total bilirubin (TB) were monitored before jaundice reduction(TBpre) and surgery within one week respectively. The efficacy of jaundice reduction in two groups was compared and further categories were carried out according to the histological types. Results TB after jaundice reduction(TBpost), TB reduction rate and the reduction adequacy rate (the percentage of postoperative TB≤100 μmol/L) in ERCP group were significantly better than those in PTCD group [(68±45)μmol/L vs (114±69)μmol/L; 73.9%±16.9% vs 64.2%±21.2%;83.8% vs 56.3%]. In patients with duodenal ampullary carcinoma,TBpost, TB reduction rate and the reduction adequacy rate in ERCP group were better than those in PTCD group [(51±20)μmol/L vs (103±49)μmol/L;78.6%±10.9% vs 67.2%±14.8%;100% vs 58.3%]. In patients with distal cholangiocarcinoma with TBpost, TB reduction rate and the reduction adequacy rate of ERCP group were better than those of PTCD group [(76±52)μmol/L vs (161±51)μmol/L;70.6%±18.5% vs 56.5%±10.4%;82.1% vs 11.1%]. Conclusions For patients with malignant obstructive jaundice, stent placement by ERCP is recommended as the first choice for preoperative jaundice reduction, especially for those with ampler carcinoma or distal cholangiocarcinoma.

Clinical Sciences

Analysis of the incidence and related risk factors of aspirin resistance in elderly patients

ZHAO Yan-qiu, LIANG Wei, HONG Yu-e

2021, 41(7):  1035-1038. 

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Objective To investigate the incidence and related risk factors of aspirin resistance (AR) in elderly patients. Methods Clincal data of ninety elderly patients who were treated in the Second People's Hospital of Hefei from December 2016 to July 2020 were collected. They took aspirin orally for more than one year with daily dose of 100 mg. The inhibition rate of aspirin on platelet was detected by thromboelastography(TEG), and the general biochemical indexes of all patients were tested. The results were analyzed, and the inhibition rate ＜50% was recorded as AR. Chi square test was used to evaluate the relationship between AR and related factors of patients' general condition. Multivariate logistic analysis was used to analyze the independent risk factors of aspirin resistance in elderly patients. Results A total of 90 patients were included, the average age was (74.06±7.90)years old, AR was 17.78% (16/90); AR had no significant difference in age, gender, smoking, creatinine, uric acid and other factors between groups, but there were significant differences in fasting blood glucose, platelet count, low density lipoprotein (LDL) between groups (P＜0.05). Logistic analysis showed that fasting blood glucose and LDL were closely related to the occurrence of AR in elderly patients (P＜0.01, OR=3.97, 95% CI=0.889-9.714), (P＜0.05, OR=3.11, 95% CI=2.42-4.01). Conclusions Fasting blood glucose and LDL seem to be the independent risk factors of AR in elderly patients. Active intervention is necessary to implement.

Tendency and quantification of related factors of regional cerebral oxygenation during laparoscopic surgery in gynecology

WANG Wei-jia, ZHANG Ning-chen, ZHANG Yue-lun, GONG Ya-hong

2021, 41(7):  1039-1042. 

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Objective To identify the time course of regional cerebral oxygen saturation (rScO₂) during laparoscopic surgery with trendelenburg position in gynecology and to quantify the effect of related factors. Methods Data from patients scheduled for elective gynecological laparoscopic surgery were collected and analyzed from June 2017 to Aug 2018 in Peking Union Medical College Hospital. Spline and mixed-effects models were used to investigate the changing trend and affecting factors of rScO₂. Results Forty-eight ASA Ⅰ~Ⅱ patients were included. rScO₂ presented a U-shaped trend over time with the spline model (P＜0.01 for the non-linear association). Episodes of cerebral deoxygenation were observed in 3 patients during the declining period of rScO₂. Mean blood pressure, systolic blood pressure and end-tidal CO₂ (ETCO₂) were positively associated with rScO₂ (P＜0.05). Peak airway pressure (PAP) and body mass index (BMI) were negatively associated with rScO₂ (P＜0.05). Compared with other factors, the effects of BMI and ETCO₂ on rScO₂ were increased significantly over time(P＜0.05). Conclusions During the gynecological laparoscopic surgery with trendelenburg position, rScO₂ changes significantly over time. BMI and ETCO₂ exhibite greater effects on rScO₂, and the effects increase significantly over time.

Case Report

A 79-year-old patient with advanced gastric cancer who received comprehensive therapy

SU Peng-fei, JIANG Lin, ZHANG Ying-jing, GUO Bo, MA Zhi-qiang, ZHOU Wei-xun, YU Jian-chun

2021, 41(7):  1043-1046. 

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Objective To discuss the current situation and challenge to the treatment strategy for elderly patients with advanced gastric cancer and provide reference for the exploration of ideal treatment methods. Methods Collect the clinical material of an elderly patient with advanced gastric cancer, and the relevant literatures were reviewed. Results A 79-year-old man was clinically diagnosed as adenocarcinoma on the fundus of stomach (cT4a-bN2Mx). After the third course of neo-adjuvant concurrent chemo-radiotherapy, a remarkable reduction and down-staging was achieved (cT3-4N1Mx), then followed by radical gastrectomy for gastric cancer. S1 was used alone for the postoperative chemotherapy. The patient received nutritional therapy in the full treatment course, and the physical condition was satisfactory without tumor metastasis and relapse two years and a half after the surgery. Conclusions The sequentially comprehensive therapy suggestes a treatment therapy for elderly patients with advanced gastric cancer, and deserves further verification and promotion clinically.

Mini Reviews

Research advances in NETs and acute respiratory distress syndrome

WANG Hang-hang, WANG Dao-xin

2021, 41(7):  1047-1051. 

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Neutrophil extracellular traps (NETs) are novel antibacterial method, which in overdose may directly or indirectly cause lung injury and promote inflammatory responses. NETs are closely related to the progression of acute respiratory distress syndrome (ARDS) and may provide new clues for the treatment of ARDS.

Research progress of inflammatory response of adipose tissue

SHU Feng, WANG Meng-yi, ZHANG Shi-wu, LI Qian-zhu, ZHANG Wei-hua

2021, 41(7):  1052-1055. 

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This review reported lipid mediators associated with inflammatory responses in adipose tissue, pro-inflammatory mediators: eicosane compounds derived from cyclooxy-genase (e.g. prostaglandins); Pro-resolving mediators: lipoxins produced by omega-6 polyunsaturated fatty acids, and resolvins produced by omega-3 polyunsaturated fatty acids. The treatment of inflammatory responses should not be limited to the use of acute cascade antagonists or inhibitors, but should be expanded to take into account the great therapeutic potential of inducers or agonists in the inflammatory resolution phase.

Research progress of mesenchymal stem cells in the treatment of pulmonary fibrosis

CHEN Min-jia, WANG Xiao-yue, CHEN Ke-hong, LU Hong-xiang

2021, 41(7):  1056-1059. 

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There is no effective therapeutic drug and intervention method for pulmonary fibrosis(PF). There are many mechanisms of using mesenchymal stem cell in the treatment of idiopathic pulmonary fibrosis(IPF), radiation pulmonary fibrosis(RPF) and other types of PF as follows: paracrine impact, release of leukocyte mediator-1 receptor antagonist, inhibition of the expression of interleukin-1 and tumor necrosis factor-α, and resistant to tissue inflammation,increase of signal transduction for hepatocyte growth factor(HGF),inhibition of PI3K/AKT/mTOR activation and epithelial mesenchymal transformation(EMT) in alveolar Ⅱ epithelial cells(ATⅡ),lung tissue apoptosis and fibrosis, promotion of tissue anti-inflammatory and anti-fibrosis, and to delay organ fibrosis. This review may function as a potential orientation for the treatment of organ fibrosis with MSCs.

Advances in mechanism of nicotine addiction

JIAO Yang, JIA Ying, ZHU Ping, ZHANG Qian, MA Xiao-li

2021, 41(7):  1060-1065. 

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Nicotine(NIC) plays a role in the brain through nicotinic acetylcholine receptors(nAChRs). nAChRs are ligand-gated ion channel composed of five transmembrane subunits. Mammals contain nine α subunits (α2～α10) and three β subunits (β2～β4). The α4 and the nAChRs subtype of β2 subunit (α4β2-nAChR), in brain tissue mediate many behaviors related to NIC addiction. Recent studies have shown that α7, α5 and α6-nAChR play an important role in regulating NIC addiction. Genetic variations in the CHRNA5-CHRNA3-CHRNB4 gene clusters encoding α5, α3, and β4 increase the susceptibility to tobacco dependence and smoking-related diseases (including lung cancer). The expression of α5 or β4 subunits in mouse brain tissue changes the consumption pattern of NIC. In addition, the effects of NIC on appetite, attention and emotion are related to the establishment and maintenance of smoking habits.

Progress in diagnosis and treatment of cystic pheochromocytoma

YANG Ting-kai, WEN Jin, JI Zhi-gang, DONG De-xin, YE Zi-xing

2021, 41(7):  1066-1070. 

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Cystic pheochromocytoma is an uncommon neuroendocrine tumor, which mostly has no obvious clinical symptoms, and difficult to make correct diagnosis before surgical operation. When a cystic mass in the adrenal gland is found by imaging examination, the possibility of cystic pheochromocytoma should be screened. At present, the most curative treatment is surgical resection. Laparoscopic adrenalectomy is the first choice. Adequate preoperative preparation and strict postoperative monitoring are important pre-requisites for ensuring the safety of patients during the peri-operative period.

Medical Education

Cultivating practical literacy of machine learning for medical students

LIU Da-lu, LI Jing

2021, 41(7):  1071-1075. 

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Machine learning (ML) is rapidly evolving into clinical artificial intelligence (AI) systems that can assist specific medical decisions. However, at present, ML faces the limitation factors such as high heterogeneity of clinical data and difficulty in data analysis for facilitating clinical performance. Future, “intelligent” doctors should dominate innovations in medicine and AI engineering around clinical mission and clinical data. Therefore, in the form of medical student internship or resident training case learning, adding practical training such as estimation of the minimum sample size required for ML and clinical interpretation of advanced characteristics of data is conducive to cultivating medical students with clinical capacity of reasoning to practice of computer-based diagnosis and treatment, and to develope a more efficient intelligent medical decision-making system.

Current situation investigation and analysis of surgical teaching for residents of Chinese ophthalmology

CHEN Di, YU Wei-hong, ZHANG Xiao, GAO Fei, WU Chan, CHEN Huan, ZHANG Mei-fen

2021, 41(7):  1076-1080. 

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Objective To investigate the current situation of surgical teaching for residents of Chinese ophthalmology. Methods Questionnaires were collected from ophthalmology teaching hospitals and standardized training centers across China. Results A total of 238 ophthalmologists were enrolled, of which 194(81.51%) participated in surgical teaching. Cataract surgery was most frequently taught (51.03%). The median number (interquartile range) of surgical operation that residents were scheduled to observe was 135(450), the number of operations that residents performed at some steps was 30(50), and the number of surgical operation that residents may perform independently was 5(19). The three tasks that teachers considered most difficult were to establish a good Wet Lab program (56.19%), the prevention of complications (51.55%) and the overall work of surgical teaching (50.52%). The response with the highest proportion of “very difficult” was to convince patients about the involvement of residents in the treatment(36.60%). The most difficult step in cataract surgery teaching was nucleus fragmentation. Conclusions Although residents' surgical training is widely carried out in China, many difficulties still exist. The number of the operation that can independently be completed by the residents is relatively low.

Investigation of study, career development and mental status of anesthesia postgraduate during the COVID-19 pandemic

YUAN Qing, LI Xu, RUAN Xia, CUI Xu-lei, TAN Gang, HUANG Yu-guang

2021, 41(7):  1081-1083. 

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Objective To investigate the training, career development and mental status of anesthesia postgraduates during the pandemic of COVID-19. Methods A self-designed questionnaire was distributed through WeChat platform to anesthesia postgraduates in Beijing, Shanghai and Chengdu. The data were recorded automatically. Results A total of 222 anesthesia postgraduates participated in the survey. Among them, 52.7% of the students were contented with outcomes of online learning, 37.84% of the students considered their graduation were negatively affected by the pandemic, 76.92% of the students thought they were more used to interview in real life than online interview, and 54.95% of the students had psychological counseling service provided by school experts during the pandemic. Conclusions During the COVID-19 pandemic, medical school endeavored to guarantee the study, career development and mental health of postgraduates.

Investigation of anxiety, depression status and analysis of related factors to clinical postgraduates during the COVID-19 pandemic

MA Ming-sheng, ZHANG Tian-nan, JIA Xue-yan, LUO Lin-zhi

2021, 41(7):  1084-1086. 

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Objective To investigate the anxiety and depression status of clinical postgraduates who returned to classes after the pandemic of corona virus disease 2019(COVID-19) and identify the correlated factors. Methods A sample of 303 clinical postgraduates who returned to school responded to an online questionnaire, including the GAD-7, PHQ-9 and 7 risk factors they worrying about. Results The overall incidence of anxiety and depression were 45.87％ and 45.54％, respectively. The results of multivariate analysis showed that worrying about “getting COVID-19” “academic writing” “delaying graduation” and “employment” was associated with the likeliness of anxiety and depression. Conclusions Clinical postgraduates still have anxiety and depression when they come back to school after the pandemic. Colleges are urged to provide psychological intervention, which is effective to alleviate their worry.

Application of blending teaching method in medical parasitology course

Rebiya·NULI, Zaihaibiya·AIHEMAITI, LI Rui, ZHANG Bing, SHAN Jiao-yu

2021, 41(7):  1087-1090. 

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Objective To study the implementation and assess effect of the “rain classroom” blending teaching method in the medical parasitology course for students of department of Traditional Chinese Medicine. Methods The 2017th grade students of department of Traditional Chinese Medicine from Xinjiang Medical University were selected. The traditional methods was used in the control group; the “rain classroom” blending teaching was implemented in the experimental group. The final comprehensive scores were compared. Results There was a significant difference in the comprehensive scores between the two groups (P＜0.05). Pass rate of the experimental group was significantly higher than that of the control group(P＜0.05). Conclusions The “rain classroom” blending teaching is conducive to improve students'achievement, self-learning capacity and curiosity, so and it is worthy to be applied to support the learning of medical parasitology.