Table of Content

05 August 2022, Volume 42 Issue 8

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Original Articles

Construction of a TCM and Western combination model for prognostic evaluation of chronic heart failure based on TCM syndrome elements and machine learning

FAN Jia-sai, DU Yi-fei, XU Jia-ying, CHEN Si-zhen, GAO Yong-hui, REN Jing-yi

2022, 42(8):  1169-1175.  doi:10.16352/j.issn.1001-6325.2022.08.1169

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Objective To construct a prognostic model of chronic heart failure (CHF) by traditional Chinese medicine (TCM) syndrome elements by machine learning method. Methods Patients with CHF admitted to the Department of Cardiology of China-Japan Friendship Hospital from January 1, 2018 to April 30, 2021 were included, and their demographic data, vital signs, co-morbidities, laboratory tests, echo-cardiographic indicators, TCM syndrome elements and treatment information were collected. The primary end point for this analysis was a model to predict cardiovascular death or hospitalization because of heart failure in one year follow-up. Least absolute shrinkage, selection operator(LASSO) regression and Cox multivariate analysis were used to screen independent risk factors that potentially affect the prognosis of CHF. A nomogram was used to establish a risk prediction model based on TCM syndrome elements. Results Totally 164 patients with an average age of (72.23±14.16) years old and 37.2% male were included in this study. The LASSO screened 9 factors from clinical variables, including coronary heart disease, hypertension, uric acid, N-terminal pro-B type natriuretic peptide (NT-proBNP), left ventricular ejection fraction (LVEF), creatine kinase-myocardial band, myoglobin, Qi deficiency and Yin deficiency. Cox multivariate regression analysis showed that Qi deficiency, hypertension, coronary heart disease, NT-proBNP and LVEF were associated with prognosis in patients with CHF. Conclusions Qi deficiency was an independent predictor of cardiovascular death or heart failure readmission in CHF patients within 1 year. The prognostic model of CHF with integrated Chinese and Western medicine has demonstrated a high accuracy.

Effect of PTOV1 expression on prognosis of colorectal cancer patients

XIE Si-an, CHEN Yi-yang, XU Jun-xuan, NING Ting-ting, ZHANG Nan, ZHU Sheng-tao, ZHANG Shu-tian

2022, 42(8):  1176-1181.  doi:10.16352/j.issn.1001-6325.2022.08.1176

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Objective To analyze the effect of over-expression of prostate tumor 1 (PTOV1) on the prognosis of colorectal cancer (CRC) based on bioinformatics, and to demonstrate the effect of PTOV1 on the proliferation of CRC cells. Methods Combined with bioinformatics and biological experiments, PTOV1 expression in CRC tissues and cell lines was analyzed. PTOV1 expression was confirmed by Western blot and RT-qPCR. The effect of PTOV1 on the proliferation of CRC cells was investigated by colony formation assay and EdU assay after transfection by PTOV1 siRNA in SW480 cell line. Results PTOV1 expression was significantly up-regulated in CRC tissues and cell lines (P＜0.05). More biological experiments results showed that PTOV1 as an oncogene was associated with poor prognosis of CRC. In vitro experiments showed that PTOV1 knockdown inhibited the proliferation of SW480 cells(P＜0.05). Conclusions PTOV1 is highly expressed in CRC and is associated with poor prognosis in CRC patients. PTOV1 is a potential prognostic biomarker for the progression of CRC.

miR-486 is potentially associated with the occurrence of OSAHS comlicated with secondary polycythemia

SHI Xue-feng, SUN Ze-rui, HE Xiang, JIE You-bang, GU Yu-hai, DUO Jie

2022, 42(8):  1182-1187.  doi:10.16352/j.issn.1001-6325.2022.08.1182

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Objective To explore the regulation and clinical significance of miR-486 in patients with sleep apnea-hypopnea syndrome (OSAHS) and secondary polycythemia. Methods Twenty-five healthy subjects who visited Outpatient Department and Health Examination Center of Qinghai Provincial People's Hospital from May 2020 to April 2021 were elected as the control group, and thirty cases of OSAHS patients with secondary polycythemia diagnosed by polysomnography and laboratory examination were selected as the OSAHS+SP group. Peripheral blood mononuclear cell (PBMC) miR-486, Sirt1 and GATA-1 expression were measured by RT-qPCR, and the concen- tration of plasma Sirt1 was measured by ELISA. In addition, RT-qPCR was used for the miR-486,Sirt1 or expression in K562 cells after the IH treatment, miR-486 over-expression or miR-486 inhibition. The Sirt1 expression was measured by RT-qPCR and Western blot. GATA-1 mRNA expression was checked by RT-qPCR in Sirt1 inhibited K562 cells. Results 1)Body mass index, red blood cells, hemoglobin (HB) and sleep apnea index(AHI) in OSAHS+SP group were significantly higher than those of control group(P＜0.001); 2)The expression of miR-486, Sirt1, and GATA-1 were significantly increased and plasma Sirt1 concentration was significantly decreased in OSAHS+SP patients PBMC(P＜0.001); 3)PBMC miR-486 and GATA-1 expression were positively correlated with AHI and HB(P＜0.05). PBMC Sirt1 expression and plasma Sirt1 level were negatively correlated with plasma Sirt1 concentration in OSAHS+SP group(P＜0.05); 4)IH promoted the expression of miR-486(P＜0.01) and inhibited Sirt1 expression in K562 cells(P＜0.01). RT-qPCR and Western blot showed that over-expression of miR-486 decreased Sirt1 expression(P＜0.01), whereas miR-486 inhibition increased the expression of Sirt1(P＜0.001). The GATA-1 expression was increased after the inhibition of Sirt1(P＜0.01). Conclusions miR-486 regulates the biological process of OSAHS patients with secondary polycythemia by targeting at Sirt1.

Correlation between serum miR-21 expression and T cells in elderly dialysis patients

WANG Shan, YANG Hua-yu, SUN Ying, MA Qing

2022, 42(8):  1188-1193.  doi:10.16352/j.issn.1001-6325.2022.08.1188

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Objective To investigate the correlation between miR-21 and peripheral blood lymphocyte subsets in elderly dialysis patients. Methods Twenty-eight elderly maintenance hemodialysis patients were enrolled as the observation group from the Department of Geriatrics of Beijing Friendship Hospital from January 2020 to January 2021. Forty-seven elderly healthy check-up people were included in the control group. The percentage of T lymphocyte subsets and the expression levels of miR-21, miR-23a, miR-150, miR-100; miR-128 and miR-211 were examined in control group and observation group. The percentage of CD4⁺ and Treg cells and the expression levels of miR-21 were detected before and after dialysis. Results The levels of systolic blood pressure, urea nitrogen, neutrophile percentage and interleukin-6 in patients with the observation group were significantly higher than that of the control group (P＜0.001). The levels of estimated glomerular filtration rate, hemoglobin, glutamic-pyruvate transaminase, albumin were significantly lower in the observation group(P＜0.001), and the percentages of CD4⁺ and Treg cells were significantly increased(P＜0.001). The relative expression of miR-21-5p was increased in the observation group(P＜0.05). Pearson correlation analysis showed that the relative expression of miR-21 was positively correlated with the ratio of Treg cells in the observation group before and after dialysis (P＜0.05). Conclusions There are micro-inflammation in elderly patients undergoing dialysis and the expression level of miR-21 in serum is positively correlated with the ratio of Treg cells before and after dialysis.

Effect of 5-Aza-2'-deoxycytidine on the expression of Sfrp1 in NRK-52E cells cultured with high glucose

TIAN Ping-ping, NING Jie, ZOU Qin, LU Yu-wei, GUO Bing, SHI Ming-jun

2022, 42(8):  1194-1199.  doi:10.16352/j.issn.1001-6325.2022.08.1194

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Objective To detect expression changes of Sfrp1 in NRK-52E cells after 5-Aza-2'deoxycytidine (5-Aza-CdR) intervention. Methods Sprague-Dawley (SD) rats were divided into normal group (NC group) and diabetic nephropathy group (DN group) randomly. The related biochemical indexes were detected after successfully modeling^[1-2] for 10 weeks;kidney index (KI) was calculated; the expression of Sfrp1 in renal tissue was detected by immunohistochemistry. NRK-52E cells were divided into normal glucose group (NG, 5.5 mmol/L), high glucose group (HG, 30 mmol/L) and high glucose +5-Aza-CdR group(5-Aza-CdR group);the expressions of Sfrp1,Dnmt3a,Dnmt3b,col-Ⅲ and col-Ⅳ were detected by Western blot;the mRNA expression of Sfrp1 was detected by RT-qPCR;pearson test was used to analyze the correlation between Sfrp1 and Dnmt3a,Dnmt3b,col-Ⅲ or col-Ⅳ. Results The level of KI, blood glucose, 24 h urinary protein and creatinine in DN group were all higher than those in NC group.The expression of Sfrp1 in renal tissue of DN group was significantly lower than that of NC group.The results of in vitro experiments showed as follows:the mRNA and protein expression of Sfrp1 were decreased in HG group(P＜0.05),while the protein expressions of Dnmt3a,Dnmt3b,col-Ⅲ and col-Ⅳ in HG group were increased (P＜0.05);compared with HG group,the mRNA and protein expression of Sfrp1 was increased in 5-Aza-CdR group(P＜0.05), the protein expressions of Dnmt3a,Dnmt3b,col-Ⅲ and col-Ⅳ in 5-Aza-CdR group were decreased (P＜0.05); Sfrp1 was negatively correlated with Dnmt3a (r=-0.937, P＜0.05),Dnmt3b (r=-0.965, P＜0.05),col-Ⅲ(r=-0.694, P＜0.05) and col-Ⅳ(r=-0.888, P＜0.05). Conclusions The mechanism through which 5-Aza-2′-deoxycytidine inhibits renal fibrosis is potentially related to the reduction of Dnmt3a and Dnmt3b expression and the increase of Sfrp1 expression.

Curcumol inhibits proliferation and promotes apoptosis of human osteosarcoma cell lines

JIANG Ning, GUO Jun, LIU Cheng, ZHOU Ju

2022, 42(8):  1200-1205.  doi:10.16352/j.issn.1001-6325.2022.08.1200

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Objective To explore the effects and mechanism of curcumol on in vitro proliferation and apoptosis of human osteosarcoma cell lines MG-63 and U2OS. Methods MG-63 cells and U2OS cells were divided into control group and 20, 40, 80 and 160 μmol/L curcumol groups. MTT assay was used to detect the proliferation levels of MG-63 and U2OS cells, and Transwell chamber assay was used for cell migration and invasion and flow cytometry was used for cell apoptosis. RT-qPCR and Western blot were applied to detect the mRNA and protein expressions of apoptosis-related proteins Bax, Bcl2, caspase and β-catenin. Results Curcumol inhibited the proliferation of MG-63 and U2OS cells in a concentration-dependent manner, and the IC₅₀ values were 128.03 μmol/L and 117.95 μmol/L respectively. Compared with the control group, curcumol significantly inhibited the in vitro migration and invasion of MG-63 and U2OS cells (P＜0.05) and induced the cell apoptosis (P＜0.05). Compared with the control group, expressions of Bax and caspase 3 were promoted while expression of Bcl2 was inhibited in curcumol groups (P＜0.05). The mRNA and protein expressions of β-catenin in MG-63 and U2OS cells were signifi- cantly inhibited (P＜0.05). Conclusions Curcumol significantly inhibits proliferation and induces apoptosis of human osteosarcoma cell lines MG-63 and U2OS.

Sevoflurane alleviates early brain injury in rats with subarachnoid hemorrhage

ZHANG Min, XING Dan-dan, KANG Wen-yue, LIN Hui

2022, 42(8):  1206-1212.  doi:10.16352/j.issn.1001-6325.2022.08.1206

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Objective To explore the protective mechanism of sevoflurane (Sev) on early brain injury (EBI) in rats with subarachnoid hemorrhage (SAH). Methods Rats were randomly divided into sham operation group (sham group), model group , Sev low dose(Sev-L, 1.5%)and high dose (Sev-H, 3%) groups, Sev+Wnt/β-catenin signaling pathway specific inhibitor DKK1 group (Sev 3%+DKK1 5 μg/kg) with 18 rats in each. After 24 hours, the neurological function of rats was scored with the improved Garcia scale, the permeability of the blood-brain barrier was detected with Evans blue (EB), and the severity of SAH was evaluated with the SAH grading system; the brain tissue water content (BWC) was calculated; the neuronal apoptosis was detected with TUNEL immunofluorescence staining; the protein expression of Wnt3a, GSK-3β, p-GSK-3β, β-catenin, ZO-1, occludin and claudin-5, MMP-9, Bcl-2, Bax in brain tissue was detected with Western blot. Results Compared with the sham group, the neurological score, expression of Bcl-2, occludin, ZO-1, claudin-5, Wnt3a, and β-catenin in brain tissues in the model group reduced significantly (P＜0.05), the SAH score, EB exudation, cell apoptosis rate, expression of Bax, MMP-9, and p-GSK-3β/GSK-3β ratio in brain tissue increased significantly(P＜0.05); compared with model group, the neurological score, expression of Bcl-2, occludin, ZO-1, claudin-5, Wnt3a, and β-catenin in brain tissues in the Sev-L group and Sev-H group increased significantly(P＜0.05), the SAH score, EB exudation, cell apoptosis rate, expression of Bax, MMP-9, and p-GSK-3β/GSK-3β ratio in brain tissue reduced significantly (P＜0.05); DKK1 significantly weakened protective effect of Sev on EBI after SAH. Conclusions Sev may activate Wnt/β-catenin pathway, improve BBB, inhibit neuronal apoptosis and reduce EBI after SAH.

Gut epithelial Depdc5/mTORC1 signaling axis regulates mouse intestinal epithelial homeostasis

ZHANG Xin-ge, MA Jie, WANG Qing-zhi, XIN Yue, YANG Chen-yan, XIONG Xi-wen

2022, 42(8):  1213-1219.  doi:10.16352/j.issn.1001-6325.2022.08.1213

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Objective To investigate the effect of over-activation of mTORC1 on mouse intestinal epithelial homeostasis by conditional knockout of Depdc5. Methods Mice were divided into Depdc5^flox/flox group (control group), Depdc5^flox/flox:Villin-Cre group (IKO group), rapamycin-Depdc5^flox/flox group (rap-Ctrl group) rapamycin-Depdc5^flox/flox:Villin-Cre group (rap-IKO group). Western blot was used to detect the protein expression of DEPDC5 and the phosphorylation S6 (PS6) which is the downstream molecule of mTORC1 in the intestinal epithelium. qPCR was applied to detect the mRNA levels of Dclk1, Trpm5 and Muc2 in intestinal epithelial cells (IECs). Hematoxylin-eosin (HE) staining was used to observe the intestinal morphology changes. Immunohistochemical (IHC) staining and alcian blue staining were used to detect the number of tuft cells, Paneth cells, proliferative cells and goblet cells. Results The protein expression of PS6 in the small intestine and colon from the mice of IKO group was obviously up-regulated than that of the control group (0.957±0.028 vs. 0.598±0.041) (P＜0.05). The mRNA levels of Dclk1, Trpm5, and Muc2 were significantly decreased in the IKO group than those in the control group (P＜0.05). The numbers of tuft cells, Paneth cells and goblet cells in the IKO group were all significantly reduced than that in the control group. Nevertheless, the crypt depth in the IKO group was significantly higher than that of the control group (P＜0.05), and the number of proliferative cells was also increased in the IKO group(P＜0.05). Conclusions The over-activation of mTORC1 inhibits the differentiation of IECs and impairs the homeostasis of intestinal epithelial.

Yuzhi Zhixue granules reduce endometrial fibrosis in rats with intrauterine adhesion

JIANG Yan-ling, DING Li, LI Fa-yu, JIE Shi-lei, ZHANG Yi-qiong

2022, 42(8):  1220-1224.  doi:10.16352/j.issn.1001-6325.2022.08.1220

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Objective To investigate the effects of Yuzhi Zhixue granules on stromal cell-derived factor-1 (SDF-1)/CXC chemokine receptor 4 (CXCR4) axis and endometrial fibrosis in rats with intrauterine adhesions. Methods The rats were randomly divided into sham operation group, model group and Yuzhi Zhixue granules low, medium and high dose groups(to establish a rat model of intrauterine adhesions), and were given 30 mg/kg, 60 mg/kg and 120 mg/kg by gavage respectively, once a day for 21 consecutive days. The levels of serum interleukin-6(IL-6), tumor necrosis factor-α (TNF-α) and transforming growth factor-β1 (TGF-β1) were detected by kits; HE staining and Masson staining were used to detect the number of endometrial glands and the degree of interstitial fibrosis; the protein expressions of SDF-1 and CXCR4 were detected by Western blot. Results Compared with those in the sham operation group, the uterine tissue structure of the model group was destroyed, the number of glands decreased, fibrotic hyperplasia was found and a large number of inflammatory cells infiltration was found. The levels of serum IL-6, TNF-α and TGF-β1 were significantly higher (P＜0.05) and the expression of SDF-1 protein in uterine tissue was significantly lower (P＜0.05). Compared with those in the model group, the number of glands in the low, medium and high dose groups of Yuzhi Zhixue Granules increased in turn, the degree of fibrosis was weakened, and the number of inflammatory cells was decreased, the levels of IL-6, TNF-α and TGF-β1 in serum were significantly lower (P＜0.05). The expression of SDF-1 protein in uterus was significantly lower (P＜0.05), and all of them were dose-dependent. Conclusions Yuzhi Zhixue granules can repair the uterine structure of rats with intrauterine adhesions, alleviate tissue fibrosis and reduce inflammatory reaction by inhibition of SDF-1/CXCR4 axis.

miR-223 inhibits pulmonary lesions of rats with pulmonary tuberculosis

TONG Xiao-wei, XIAO Han

2022, 42(8):  1225-1229.  doi:10.16352/j.issn.1001-6325.2022.08.1225

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Objective To explore the effect of miR-223 on pulmonary lesions and IFN-γ in rats with pulmonary tuberculosis(PT) mechanism of protein expression. Methods Rats were divided into normal group (NC group), pulmonary tuberculosis rat model group (PT group) and pulmonary tuberculosis rat model transfected with miR-223 mimics group (miR-223 group) with 10 in each group. T cells in peripheral blood was quantified by flow cytometry, IFN-γ in serum was detected by ELISA and IL-18 levels, and the number of Mycobacterium tuberculosis colonies was counted; the pathological changes of lung tissue were micros copied by HE staining; IFN-γ in lung tissue was detected by immunohistochemistry method; RT-qPCR was used to detect miR-223 mRNA and IFN-γ in lung tissue MRNA expression. Results In PT group, the peripheral blood T cell subsets were disordered, the level of CD3⁺,CD4⁺% significantly decreased, and CD8⁺% was increased (P＜0.05); CD3⁺ and CD4⁺ percentage in miR-223 group were higher than those in PT group, while the percentage of CD8⁺ was lower(P＜0.05). Serum IFN-γ and IL-18 in PT group increased(P＜0.05); the number of Mycobacterium tuberculosis colonies in PT group was more than that in NC group (P＜0.05); the colony number of Mycobacterium tuberculosis in miR-223 group was significantly less than that in PT group (P＜0.05). Total IFN-γ level in PT group was higher than that in NC group(P＜0.05) but was significantly inhibited in miR-223 group(P＜0.05). Conclusions miR-223 can inhibit the pulmonary inflammatory response caused by Mycobacterium tuberculosis, and miR-223 showed an effect of anti-tuberculosis bacteria and may regulate host immunity, so and can effectively inhibit the expression of IFN-γ in lung tissue and thus alleviates severity of tuberculosis.

miR-499 inhibits migration and promotes apoptosis of human osteosarcoma cell line Saos2 by targeting at TGF-α

WANG Lei, QIU Ming-xian, ZHANG Hui-rong, ZHANG Jin-ping, ZHAO Jing, KANG Xiao, ZHANG Qing-quan

2022, 42(8):  1230-1236.  doi:10.16352/j.issn.1001-6325.2022.08.1230

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Objective To explore the role of miR-499 targeting TGF-α mRNA protein in regulating the migration and apoptosis of human osteosarcoma cell line Saos2 and its pathogenesis. Methods The expression differences of miR-499 and TGF-α in osteosarcoma tissues were detected by RT-qPCR. Double luciferase reporter gene assay was used to detect the targeting relationship between miR-499 and TGF-α. Transwell cell assay and TUNEL apoptosis assay were used to detect the migration and apoptosis of Saos2 cells. A nude mouse xenograft model of osteosarcoma was constructed to detect the regulatory mechanism of miR-499/TGF-α axis in vivo. Results In osteosarcoma, miR-499 expression was decreased (P＜0.01) but TGF-α mRNA expression was increased(P＜0.01). miR-499 negatively regulated TGF-α mRNA protein expression (P＜0.01). Transfection of miR-499 simulacrum inhibited the migration and promoted apoptosis of Saos2 cells (P＜0.01), while TGF-α promoted the migration and inhibited apoptosis of Saos2 cells (P＜0.01). miR-499 inhibited the carcinogenic effect of TGF-α. Conclusions miR-499 inhibits the biological function of osteosarcoma cell line Saos2 by inhibiting at TGF-α mRNA protein level which suggests that miR-499 may inhibit the development of osteosarcoma.

Esculin inhibits the proliferation of human nasopharyngeal carcinoma cell line HNE-3

SHEN Yong-hua, WANG Jun-feng, CHENG Ze-xing, FENG Juan-juan

2022, 42(8):  1237-1242.  doi:10.16352/j.issn.1001-6325.2022.08.1237

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Objective To explore the effect of esculin on the biological behavior of nasopharyngeal carcinoma cells and its possible mechanism. Methods Different concentrations of esculin were used to treat human nasopharyngeal carcinoma cells HNE-3, si-NC, si-circ-ZNF609 were transfected into HNE-3 cells, pcDNA, pcDNA-circ-ZNF609 were respectively transfected into HNE-3 cells and then treated with esculin. CCK-8 experiment was used to detect cell proliferation. Intracellular glucose uptake levels and lactate levels were tested using kits. The activity of pyruvate kinase and hexokinase was tested using an enzyme activity kit. The Transwell chamber test was used to find cell migration and invasion. RT-qPCR method was used to detect the expression of circ-ZNF609. Protein expression of MMP-2 and MMP-9 was calculated by Western blot. Results Esculin could reduce cell survival rate and the protein levels of MMP-2, MMP-9 in a concentration-dependent manner (P＜0.001), could reduce glucose uptake and lactate levels (P＜0.001), and may reduce pyruvate kinase and the activity of hexose kinase (P＜0.001), could also reduce the number of migrating and invading cells (P＜0.001). Esculin could reduce the expression of circ-ZNF609 (P＜0.001), and it was dose-dependent. After transfection with si-circ-ZNF609, the cell survival rate and the protein levels of MMP-2, MMP-9 were decreased (P＜0.001), glucose uptake and lactate levels were decreased(P＜0.001). The activity of pyruvate kinase and hexokinase and the number of migrating and invasive cells were all decreased (P＜0.001). Transfection of pcDNA-circ-ZNF609 may restore the inhibitory effects of esculin on the proliferation, glycolysis, migration and invasion of HNE-3 cells. Conclusions Esculin could inhibit cell proliferation, glycolysis, migration and invasion in HNE-3 cells by regulating the expression of circ-ZNF609.

Analysis of cyclophosphamide induced ovarian insufficiency and differentially expressed genes in rats

ZHANG Yu-lin, ZOU Heng, ZHANG Chan-yu

2022, 42(8):  1243-1249.  doi:10.16352/j.issn.1001-6325.2022.08.1243

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Objective To establish a stable cyclophosphamide(CTX) induced ovarian insufficiency rat model and to explore the mechanisms. Methods The survival curve of rats after intraperitoneal injection of different dosage of CTX was drawn and the optimal dose was determined. Sex hormone level, estrous cycle and ovarian histological change were further observed. The underlying mechanisms were explored by transcriptome sequencing. Results The mortality rate of medium and high dose in CTX group was high, so low dose CTX was chosen. After CTX exposure, E2 and AMH decreased(both P＜0.05),while FSH increased(P＜0.01). About half of the model rats showed irregular estrous cycles(P＜0.05). The preantral and antral follicles decreased(P＜0.01, P＜0.05), while atresia follicles increased (P＜0.01). Transcriptome sequencing analysis showed that differentially expressed genes (DEGs) between two groups functioned in channel activity and in transmembrane transporter activity. DEGs mainly enriched in neuroactive ligand-receptor interaction pathway, cAMP signal pathway and calcium signaling pathway, axon guidance. According to the protein-protein interaction network of DEGs,we screened out 14 hub genes. Among them, Leptin, Shh,Ntrk2,GRIN2A,CAMK2A,CaSR and Pvalb have been revealed to be closely related to ovarian function. Conclusions A POI rat model induced by CTX was successfully constructed; the neuroactive ligand-receptor interaction pathway, cAMP signal pathway, calcium signaling pathway and axon guidance pathway are enriched pathways. Lep, Shh, Ntrk2, GRIN2A, CAMK2A, CaSR and Pvalb are potentially key genes.

Ambroxol hydrochloride alleviates acute lung injury induced by gastric juice aspiration in rats

CUI Yi-yao, WANG Feng, LYU Xiao-shuo, LIU Chun-quan, XU Kai-kai, CUI Yong

2022, 42(8):  1250-1254.  doi:10.16352/j.issn.1001-6325.2022.08.1250

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Objective To investigate the therapeutic and protective effect of ambroxol hydrochloride on acute lung injury caused by aspiration of gastric juice in rats. Methods SD rats were randomly divided into control group, model group (constructed by trans-cricothyroid membrane puncture with pH≈2.3 rat gastric juice, 1.5 mL/kg) and ambroxol hydrochloride intervention group (30 mg/kg). After 24 hours of modeling, the drug was administered through caudal vein once a day for 3 days. HE staining was used to detect lung tissue, lung wet to dry weight ratio (W/D) value and arterial blood gas analysis to evaluate the degree of lung injury, ELISA was used to detect IL-6, IL-10, TNF-α and C-reactive protein (CRP) levels in serum. Results Compared with the control group, the alveolar structure showed atrophy, interstitial edema, and inflammatory cell exudation. Hyaline membrane was formed and W/D value increased, pH and arterial oxygen pressure (pO₂) decreased, arterial carbon dioxide pressure (pCO₂) increased, and serum inflammatory factors (IL-6, IL-10, TNF-α and CRP) increased (P＜0.05). Compared with the model group, the intervention reduced the pathological changes of lung tissue, W/D value decreased, pH and pO₂ increased, pCO₂ decreased, serum inflammatory factors (IL-6, IL-10, TNF-α and CRP) decreased(P＜0.05). Conclusions Ambroxol hydrochloride can effectively reduce the acute lung injury in rats caused by gastric juice aspiration and has good therapeutic and protective effects.

Influence of preoperative ureteral stenting on the curative effect of flexible ureteroscopic lithotripsy

LI Xin, CHEN Zhi-gang, ZHANG Meng-dong, TANG Hao, LU Xiao-dong, LI Bao-guo

2022, 42(8):  1255-1258.  doi:10.16352/j.issn.1001-6325.2022.08.1255

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Objective To investigate the therapeutic effect of preoperative ureteral stenting on the treatment of renal or upper ureteral calculi with flexible ureteroscopic lithotripsy. Methods From October 2018 to July 2021, 256 patients who had undergone flexible ureteroscopic lithotripsy for renal or upper ureteral calculi in Beijing Haidian Hospital were retrospectively selected. According to whether the patients accepted preoperative ureteral stenting or not, the patients were divided into experimental group (n=192) and control group (n=64), and the experimental group was further divided into long-(14 d, n=64) and short-term (7 d, n=128) group according to the time of preoperative ureteral stenting. The differences in success rate of surgery, operative time, amounts of intraoperative blood loss, length of hospital-stay, incidence of postoperative complications and stone-free rate were analyzed to evaluate the therapeutic effects. Results The operative time and intraoperative blood loss of the control group were significantly higher than that of the experimental group, there was significant difference between the two groups(P＜0.001). The long-term group had longer hospital-stay time (P＜0.001), shorter operative time and less amounts of blood loss (P＜0.001). The success rate of surgery and stone-free rate of the control group were lower than that of the experimental group (P＜0.05), while incidence of postoperative complications was higher than experimental group (P＜0.001). There was no significant difference between the short- and long-term group in the stone-free rate, while the postoperative complications of the short-term group was higher than the experimental group (P＜0.05). Conclusions Preoperative ureteral stenting improves the success rate of surgery operation and the stone-free rate, reduces the postoperative complications and intraoperative blood loss, shorten the operative time.

Clinical Sciences

Effects of flexible ureteroscopy lithotripsy and percutaneous nephrolithotripsy on the efficacy and inflammatory response of renal calculi less than 3 cm

YU Zi-qiang, XU Jiu-ping, ZHAN Chang-sheng, SHAO Ming-feng, ZOU Jian-an

2022, 42(8):  1259-1262.  doi:10.16352/j.issn.1001-6325.2022.08.1259

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Objective To compare the effect of flexible ureteroscopy lithotripsy and percutaneous nephrolithotripsy in treatment of renal calculi less than 3 cm. The effectiveness, reliability and the effect on inflammatory response of the two methods were compared. Methods From January 2015 to September 2021, 74 patients with renal calculi were collected and divided into FURS group and PCNL group with 35 in FURS group and 39 in PCNL group. The indexes before and after operation were compared. Results The statistically significant difference was found in operation time [(69.6±19.0)min vs(84.8±20.5)min], hospitalization days [(3.8±1.1)d vs (6.4±1.3)], intraoperative bleeding [(21.4±8.9)mL vs (75.4±48.6)mL,P＜0.01], pain VAS score [(1.0±0.9) vs (3.2±1.6),P＜0.01],freiburg patients satisfaction [(1.7±0.8) vs (2.2±1.1),P＜0.05] and complications (2.9% vs 17.9%,P＜0.05). Three days after operation, the COR[(209.3±20.9)ng/mL vs (226.4±38.8)ng/mL P＜0.05] and MDA [(6.9±0.6)nmol/mL vs (10.0±1.0)nmol/mL,P＜0.01] in the FURS group were lower than those in the PCNL group, while SOD [(70.6±8.3)NU/mL vs (61.3±7.8)NU/m) was higher than those in the PCNL group and the difference was statistically significant. The IL-6[(11.8±1.1)pg/mL vs (14.1±1.1)pg/mL,P＜0.001), IL-10 [(9.1±0.9)pg/mL vs (12.5±1.1)pg/mL,P＜0.001] and TNF-α[(1.7±0.1)ng/mL vs (2.3±0.1)ng/mL,P＜0.001]in the FURS group were significantly lower than those in the PCNL group three days after operation. Conclusions FURS is confirmed to be a safe and effective treatment for renal calculi less than 3 cm because of with low oxidative stress and inflammatory response.

Retrospective analysis of occlusion released under guidance of echocardiography in the treatment of 54 patients with congenital heart disease

PAN Guang-yu, LIU Ruo-fan, GONG Da, LIU Shen, TONG Ming-hui, LI Bin, XU Jian-ping

2022, 42(8):  1263-1268.  doi:10.16352/j.issn.1001-6325.2022.08.1263

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Objective To share our experience of percutaneous or transthoracic closure of atrial septal defect (ASD) and ventricular septal defect (VSD) supported by echocardiography guidance. Methods From December 2010 to September 2019, 54 patients of congenital heart disease (30 cases of isolated atrial septal defect, 24 cases of ventricular septal defect) were treated with surgical operation by occluder under guidance of echocardiography closed without cardiopulmonary bypass. Fourteen cases of ASD were performed device closure through a small right sternal margin thoracotomy incision at the forth intercostal space, 16 cases of ASD underwent percutaneous occlusion surgery, and 24 cases of VSD were closed through small incision under the xiphoid process. Results The operations were successful in the 24 cases of VSD. One of the cases with minimally incision ASD occlusion was displaced 7 hours after operation, the occluder was taken out and ASD was repaired under cardiopulmonary bypass in emergency. There were 13 cases of successful occlusion and 3 cases of failure in percutaneous ASD closure patients. Among three cases, 2 cases underwent ASD closed through transthoracic closure and 1 case underwent open surgery under cardiopulmonary bypass successfully. The 46 cases were followed up for 3-103 months, with an average of (21.3±16.2)months. No occluder displacement, shedding, significant residual shunt were observed. Conclusions Echocardiography-guided percutaneous atrial septal defect and ventricular septal defect is safe and effective without radiation and CPB. Percutaneous closure of atrial septal defect is suitable for children over 3 years. Atrial septal defect and ventricular septal defect occlusion through small thoracic incision is feasible and easy to operate in other healthcare institutions.

Case Report

An unusual case of intraoperative hyperkalemia

MA Lu-lu, HUANG Yu-guang

2022, 42(8):  1272-1273.  doi:10.16352/j.issn.1001-6325.2022.08.1272

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Objective To report a rare case of intraoperative hyperkalemia and share our experience in the diagnosis and treatment with colleagues. Methods We reported 1 case of intraoperative hyperkalemia caused by central venous occlusion, auto-circulation of dialysis circuit and incomplete dialysis, the manifestation and treatment were summarized. Results Hyperkalemia is common in patients dependent on hemodialysis, and is a life-threatening complication. Electrocardiogram was not sensitive to hyperkalemia and the manifestations of intraoperative hyperkalemia are atypical, making the diagnosis difficult. After intravenous administrator of calcium and insulin,correction of acidosis and the application of continuous renal replacement therapy (CRRT), the patient started peritoneal dialysis gradually. Conclusions For patients who are dependent on hemodialysis and complicated with the failure of vascular access, incomplete dialysis is possible and peritoneal dialysis should be considered.

Mini Reviews

Research progress of RNA binding motif protein family-mediated alternative splicing in cardiomyopathy

LIU Shu, DING Hong, ZHAO Ke-xin, BAI Feng, ZHANG Xiao-wei

2022, 42(8):  1274-1278.  doi:10.16352/j.issn.1001-6325.2022.08.1274

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RNA binding motif protein (RBM) is a type of RNA binding protein, which contains RNA recognition motif, RNA binding motif and ribo-nucleo-protein motif. The alternative splicing mediated by members of the RBM family is closely related to myocardial contractile proteins, myocardial development and myocardial sodium channels, which play an important role in pathogenesis of myocardial diseases and even heart failure.

Research progress on the role and mechanism of visfatin in sepsis-related organ injury

KUANG Zhong-shen, CONG Zhu-kai, WU Chang-yi, YANG Ning, ZONG Ya-nan, GUO Xiang-yang

2022, 42(8):  1279-1283.  doi:10.16352/j.issn.1001-6325.2022.08.1279

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Sepsis is a common acute and critical disease in clinic. Systemic inflammatory response (SIRS) is one of the key mechanisms for the occurrence of sepsis. Visfatin, as an important cytokine, is involved in the occurrence and development of “cytokines storm” caused by sepsis. Recent clinical and basic research have shown that visfatin plays an important role in the occurrence and development of sepsis, and inhibition of visfatin expression can improve the systemic inflammatory response and the damage of lung, heart, liver and other important organs caused by sepsis. Based on the analysis of current clinical and basic research results, visfatin are expected to become an indicator of the severity of sepsis and a potential target for clinical intervention.

Research progress of programmed cell death in the occurrence and development of sepsis

XUE Jin-fang, SONG Hai-fei, CHEN Guo-bing

2022, 42(8):  1284-1287.  doi:10.16352/j.issn.1001-6325.2022.08.1284

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Sepsis has high morbidity and mortality in critically ill patients.The occurrence and development of sepsis will lead to programmed cell death. All kinds of programmed cell death mediate inflammatory response and oxidative stress through different signal transduction pathways, and cause irreversible damage in patients with sepsis, and finally develop into multiple organ failure. At present, programmed cell death has been found, including apoptosis, necroptosis, pyroptosis, ferroptosis and so on. The development and progression of sepsis is accompanied by various types of programmed cell death.These results of research provide new ideas for exploring the pathophysiological mechanism of sepsis and strategies of clinical treatment.

Research progress of stem cells aging

ZHU Si-lu, LUO Dan

2022, 42(8):  1288-1291.  doi:10.16352/j.issn.1001-6325.2022.08.1288

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Stem cells have broad therapeutic potential in many fields and have been used as seed cells in regenerative medicine and tissue engineering research. However, stem cell aging is a challenge to their clinical applications. By performing effective therapeutic interventions for aging stem cells, stem cells will bring greater hope for clinical practice. This review summarizes the latest advances in stem cells biology, so to delay aging of stem cells by potential intervention to the aging process. The results of research in this field will contribute to the development of new rational treatment strategies.

Research progress on the effects of mineralocorticoid receptor on cardiovascular diseases

MA Qian-yu, PENG Shi, ZHANG Zhao-yuan, ZHANG Dan, ZHANG Jin

2022, 42(8):  1292-1296.  doi:10.16352/j.issn.1001-6325.2022.08.1292

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The mineralocorticoid receptor (MR) is a ligand-activated cytoplasmic receptor.Activation of MR in different cells can cause cardiovascular inflammation, oxidative stress, tissue remodeling ,and even function change the activity of ion channels resulting in incidence of arrhythmia through various mechanisms.MR antagonists are effective medicine to treat hypertension,coronary heart disease,and heart failure. In addition, the role of MR in arrhythmia and pulmonary hypertension also provides new perspectives for future clinical medication.

Progress of artificial intelligence application in cancer diagnosis and treatment

DU Ming-yue, LI Xue-guang, ZUO Shan-ru, CHEN Zi-qi, ZHOU Jun-hua, LI Hao, HE Quan-yuan

2022, 42(8):  1297-1301.  doi:10.16352/j.issn.1001-6325.2022.08.1297

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The artificial intelligence (AI) technology with supreme capacity of big data integration and management have been used in digital diagnosis and clinical treatment,which have greatly facilitated the development of clinical oncology and precision medicine. AI technology is far superior to human beings in big data integration and analysis, so has been widely applied in pathological image analysis, molecular marker discovery, clinical decision support, drug discovery and recommendation and so on. In recent years, many remarkable achievements have been made in the field. This review discussed the latest applications of AI-assisted precision cancer diagnosis and treatment, highlighted emerging challenges, and predicted the development trend in the future.

Research progress of insulin resistance markers in atherogenesis

ZHAO Mao-yu, LI You-mei, LIU Huan-yun, MAO Qi

2022, 42(8):  1302-1305.  doi:10.16352/j.issn.1001-6325.2022.08.1302

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Insulin resistance (IR) is the pivotal link between metabolic disorders and atherosclerosis (AS). Insulin resistance plays an important role in the initiation and progression of atherosclerosis, and its underlying mechanism involves complex pathophysiological processes including endothelial injury, inflammatory reaction, and oxidative stress. Insulin resistance markers presented the potential association of metabolic risk with atherosclerosis and the detection of insulin resistance markers may support strategy development for the clinical management of cardiovascular diseases.

Advances of research on the function of RNA binding proteins in the pathogenesis of thyroid carcinoma

ZHOU Mei-cen, LAN Ling, DENG Wei

2022, 42(8):  1306-1309.  doi:10.16352/j.issn.1001-6325.2022.08.1306

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RNA-binding protein (RBP) plays an important role in the pathogenesis of thyroid cancer. It can regulate the intracellular activities such as transcription, nuclear output, alternative splicing and translation of mRNA from tumor-related genes at the post-transcriptional or translation levels, and then regulate the proliferation, apoptosis and migration of thyroid cancer, thereby promoting or inhibiting the occurrence of tumors. Clinically, a series of RBP molecules are significantly correlated with the classification and prognosis of patients with thyroid cancer and may support the oncological classification, evaluation and prediction of prognosis of thyroid cancer.

Function of the microglia in neuroinflammation of Alzheimer's disease:a brief review

WANG Yuan-zhuo, ZHENG Qing-yue, ZHANG Han-lin, QIU Wen-ying, WANG Tao, MA Chao

2022, 42(8):  1310-1313.  doi:10.16352/j.issn.1001-6325.2022.08.1310

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Different types of activated imbroglio exist in the brain of the patients with Alzheimer's disease (AD). As the neuroinflammation reaction progresses, significant changes take place in the morphology and function of those cells. They could play a pathogenic role by altering the phagocytic function of cells, including intracellular toxic reactions or directly participate in immune response. Several clinical trials have been conducted to promote targeted therapy focusing on the role that microglia played in neuroinflammation reactions of AD. More large scale studies are necessary to elucidate the function and mechanism of microglia in the neuroinflammation reaction of AD.

Medical Education

Practice of standardized training simulation examination in the national standardized training base for residents in ultrasound medicine

JIA Xin-ying, TIAN Yan, LU Wei-dan, XI Xue-hua, ZHOU Tong-tong, LU Xiao, ZHANG Bo

2022, 42(8):  1314-1317.  doi:10.16352/j.issn.1001-6325.2022.08.1314

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Objective To evaluate standardized training simulation examination for residents trainees in ultrasound medicine. Methods From 2018 to 2020, all the 17 resident trainees participated in the training at the standardized training base for residents located in the department of ultrasound medicine of China-Japan Friendship Hospital.The examination was conducted in stages for capacity building for clinical reasoning, the equipment operation and humanity, three rounds.The assessment performance was analyzed in groups based on academic degree and grade respectively. Questionnaire survey was implemented for trainees' subjective evaluation for simulation examination. Results Both clinical reasoning and the ultrasonic operation dealing with heart check by the graduate group and the undergraduate group showed a progressing trend for three grades, and the postgraduate group had higher performance than the undergraduate group {[clinical reasoning (342.10±26.73) points vs (269.55±30.35) points, (336.00±22.58) points vs (287.18±22.18) points, (352.00±31.00) points vs (327.27±22.31) points], [untrasonic operation for heart check (89.67±7.59) points vs (62.00±2.00) points, (83.00±7.79) points vs(67.50±7.50) points, (97.33±2.05) points vs (89.50±2.50) points]}. There was no statistically significant difference among grades. Performance was evident between grades 1 and 3 with the machine operation [(243.19±18.39) points vs (268.00±25.06) points]. In the undergraduate group, the 1st and 3rd examination progressed significantly [(234.59±15.24) points vs (262.00±33.34) points]. The questionnaire showed that 17 trainees believed that the simulation examination effectively improved their ultrasound skills. Conclusions The simulation examination can improve the clinical skill and increase trainees' confidence, so this method should be shared by other institutions for standardized residents training.

Medical students participate in bone and joint specimen preparation in the second classroom

SHEN Xin-hua, LIU Wei, LI Wen-ting, CAO Cheng-gang, MA Chao

2022, 42(8):  1318-1321.  doi:10.16352/j.issn.1001-6325.2022.08.1318

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Objective To evaluate the effect of participation in bone and joint specimen preparation on anatomy exam scores and medical students' feedback to the second anatomy classroom. Methods The students of “4+4” program of clinical medicine enrolled in 2021 were divided into experiment and control groups. The students in experiment group were asked to prepare joint specimens under the guidance of teachers in after class time, while the students in control group only required to observe the specimens. The anatomy exam scores of the two groups were analyzed and and questionnaire survey was implemented to collect students' feedback about the joint specimen preparation. Results There was no significant difference in the scores of final examiniation between the experiment group and the control group. However, the lowest and the highest correct rates in experimenl group (40% and 95% respectively) were both higher than that of the control group (32.5% and 82.5% respectively). If the correct rate of joint specimens was calculated separately, the experiment group (87.0%±16.7%) was significantly higher than that of control group (66.7%±24.3%) (P＜0.01). In the limb theory test, the mean score of the experiment group (84.7%±20.5%) was significantly higher than that of the control group (69.3%±14.5%) (P＜0.05). The students gave a high evaluation to the joint specimen preparation in after class time or called second classroom. Conclusions Medical students' participation in joint specimen preparation as the second classroom can improve their learning outcomes and this second classroom strategy as a supplement to the first classroom may be potentiallt shared by other medical schools.

An exploration of medical imaging teaching remodeling led by clinicians

SHAN Ying, JIN Ying, GU Yu, WANG Wei, LI Yan, PAN Ling-ya

2022, 42(8):  1322-1324.  doi:10.16352/j.issn.1001-6325.2022.08.1322

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Medical imaging plays an important role in clinical works. However, most students are challenged by difficulty to apply imaging knowledge into clinical practice, mainly due to the lack of relevant training and guidance. This study initially explored the remodeling of imaging teaching pathway led by clinicians,merge medical image learning into clinical work, before, during and after operation. The method pays more attention to clinical reasoning development and is welcomed by both undergraduates and trainees of standardized residents' training program. We recommend teaching model to other medical schools without hesitation.