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28 July 2022, Volume 42 Issue 7 Previous Issue    Next Issue
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Basic research
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Basic research

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Emergence of ciprofloxacin heteroresistance in clinical Pseudomonas aeruginosa LI Congcong, YAO Yufeng, ZHANG Chuanzhen 2022, 42 (7):  839-845.  doi: 10.3969/j.issn.1674-8115.2022.07.001 Abstract ( 424 )   HTML ( 31 )   PDF (4880KB) ( 452 )   ·To investigate ciprofloxacin heteroresistance in Pseudomonas aeruginosa (PA). ·Ciprofloxacin heteroresistance in 227 clinical PA strains was initially identified by disk diffusion. Meanwhile, susceptibilities of these strains was classified according to the diameter of the inhibition zone. The stability of the identified heteroresistant strains was tested by measuring the sensitivity after serial passage under antibiotic-free conditions. Mechanisms mediating heteroresistance were analyzed by whole-genome sequencing. Susceptibilities of resistant subpopulations to different antibiotics were also detected. ·Based on disk diffusion, 142 (62.6%), 26 (11.5%), and 59 (26.0%) of 227 isolates were classified as susceptible, intermediate and resistant to ciprofloxacin respectively. Eighteen putative heteroresistant strains were detected through primary disk diffusion selection, and 11 among them were further verified as stable heteroresistant strains by population analysis profiling (PAP). By whole-genome sequencing, 8 and 2 among 11 resistant subpopulations carried mutations in mexS and mutations in gyrA individually. In addition, mutations in fleQ, PA2632 or PAKAF_02255 were detected respectively. Both mexS and gyrA mutants led to moxifloxacin resistance, and some mexS mutants also showed resistance to chloramphenicol and imipenem. ·Ciprofloxacin heteroresistance rate inPA is 4.8% (11/227). As a screening method for heteroresistance of PA, the disk diffusion has low sensitivity and has a certain false negative rate. The mutation mexS is the predominant mutation type in resistant subpopulations and it might enhance the overexpression of efflux pump MexEF-OprN, thus mediating resistance to different antibiotics including ciprofloxacin. Figures and Tables | References | Related Articles | Metrics

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Construction of OPEI vector for silencing TRAF6 to promote cartilage regeneration in inflammatory environment LIU Hongqiang, LU Yanqing, GAO Yuxuan, WANG Yiyun, WANG Chuandong, ZHANG Xiaoling 2022, 42 (7):  846-857.  doi: 10.3969/j.issn.1674-8115.2022.07.002 Abstract ( 362 )   HTML ( 18 )   PDF (6757KB) ( 335 )   ·To construct a low toxicity and high-efficiency joint synovial siRNA transfection vector OPEI, and inhibit tumor necrosis factor receptor-associated factor 6 (TRAF6) to rescue the chondrogenic ability of bone marrow mesenchymal stem cells (BMSCs) under inflammatory conditions. ·The osteoarthritis (OA) models of SD rats (n=20) were established by medial meniscectomy of knee. Another sham operation group (n=10) was established, and the meniscus remained intact. The cartilage and synovium were collected 3 months after surgery. The expression of TRAF6 was detected by immunohistochemistry, and Western blotting was used to detect the expression of MMP13, TRAF6 and p-p65 in primary rat synovial cells induced by interleukin-1β (IL-1β). Further, the small molecule polyethylenimine (PEI) derivative OPEI was synthesized in anhydrous anaerobic environment, and the ability of OPEI to encapsulate siRNA was detected by agarose gel electrophoresis. The particle size and Zeta potential of OPEI/siRNA complex were measured by dynamic light scattering. The cytotoxicity of the formed complex to rat primary synovial cells was detected by MTT method. The effect of the complex on synovial cell apoptosis was analyzed by flow cytometry. The transfection efficiency of siRNA by OPEI in synovial cells in vivo and in vitro was detected by laser confocal technique and fluorescence microscopy. The proteoglycan content of chondrocyte matrix was detected by alcian blue staining. ·Compared with the sham operation group, TRAF6 was highly expressed in synovium and cartilage of the rat OA models, and inhibition of TRAF6 could significantly reduce the expression of MMP13 and p-p65 in IL-1β-stimulated primary synovial cells. The siRNA transfection efficiency of OPEI in the rat primary synovial cells was as high as 99.33%. A large number of synovial cells ingested siRNA on the 3rd and the 7th day after injection of OPEI / Cy3-siRNA into rat knee joints. Two days after OPEI / siTRAF6 complex with different w/w ratios was transfected into rat primary synovial cells, the results of TRAF6 protein expression showed that the knockout efficiency of TRAF6 gene in the OPEI / siTRAF6 group was 49.05%, 74.61% and 83.18% respectively when the mass ratio was 3∶1, 4∶1 and 5∶1. After TRAF6 gene was silenced by OPEI/siTRAF6, compared with the control group (OPEI/siNC), alcian blue staining of chondrocytes was significantly enhanced under IL-1β stimulation. ·OPEI is a low toxicity and high efficiency siRNA transfection vector, silencing TRAF6 in synovial cells could promote OA cartilage regeneration. Figures and Tables | References | Related Articles | Metrics

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SMAD7 expression in multiple myeloma and its effect on cell proliferation and drug resistance DING He, CAO Yanglin, HE Yang, WEI Xing, YANG Jianfeng 2022, 42 (7):  858-865.  doi: 10.3969/j.issn.1674-8115.2022.07.003 Abstract ( 429 )   HTML ( 26 )   PDF (2182KB) ( 289 )   ·To analyze the expression of Sma- and Mad-related protein 7 (SMAD7) in multiple myeloma (MM), and its effect on the proliferation and drug resistance of MM cells. ·Three datasets from the Gene Expression Omnibus (GEO) database were obtained to analyze the differential expression of SMAD7 in healthy donors and MM. Bone marrow samples from 8 healthy donors and 20 MM patients were collected. The relative expression of SMAD7 mRNA in bone marrow CD138+ cells was detected by quantitative real-time PCR (qPCR), and the relationship between SMAD7 mRNA expression and clinical information of patients was analyzed. After overexpression of SMAD7 in MM cells KMS11, the effects of SMAD7 on the proliferation was observed by CCK8 assay and cell cycle assay. KMS11 cells overexpressing SMAD7 were treated with different concentrations of bortezomib, and the effect of SMAD7 on drug resistance and apoptosis of MM cells were observed. ·The analysis of SMAD7 expression data in the datasets showed that compared with healthy donors, SMAD7 expression in MM patients was higher (all P<0.05). The results of qPCR showed that the relative expression of SMAD7 mRNAwas up-regulated in MM patients (P=0.002), although no correlation with clinical information of patients was shown. Compared with the control cells, the cell viability of KMS11 was higher after overexpression of SMAD7 (all P<0.05); the change of cell cycle distribution showed that the proportion of cells in S phase decreased (P=0.016), and the proportion of cells in G2/M phase increased (P=0.005); after bortezomib treatment, the drug sensitivity and apoptosis level of overexpression SMAD7 cells was lower. ·The expression of SMAD7 is up-regulated in MM. Overexpression of SMAD7 can promote the proliferation and drug resistance of MM cells. Figures and Tables | References | Related Articles | Metrics

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Levels of adiponectin in the peripheral blood of patients with immune thrombocytopenia and its effect on the differentiation of megakaryocyte cell line LI Xinyu, ZUO Bin, WANG Wen, NIU Xiaoyin, WENG Zhen, HE Yang 2022, 42 (7):  866-874.  doi: 10.3969/j.issn.1674-8115.2022.07.004 Abstract ( 313 )   HTML ( 17 )   PDF (2387KB) ( 338 )   ·To explore the level of adiponectin in the peripheral blood plasma of patients with immune thrombocytopenia (ITP) and its effect on the differentiation and maturation of megakaryocytes. ·From January 2021 to February 2022, the peripheral blood samples were collected from 46 ITP patients in the Department of Hematology, The First Affiliated Hospital of Soochow University and 30 healthy controls with corresponding gender and age in the physical examination center. The plasma adiponectin levels in the ITP patients and healthy control (HC) group were detected by using human adiponectin enzyme-linked immunosorbent assay (ELISA) kits. And the correlation between adiponectin level and body mass index (BMI) in the ITP patients was analyzed. In terms of cell experiments, real-time quantitative PCR (RT-qPCR) and Western blotting were used to study the expression of adiponectin receptors (ADIPOR1 and ADIPOR2) in the myeloid cell line K562, and the megakaryocyte cell lines MEG-01 and Dami at mRNA and protein levels, respectively. The differentiation of K562 cell line and the maturation of MEG-01 and Dami cell lines were induced by phorbol 12-myristate 13-acetate (PMA). At the same time, the cells were treated with different concentrations of adiponectin receptor agonist AdipoRon. Flow cytometry was used to detect the percentage of CD41+ cells, the mean fluorescence intensity (MFI) of CD41, and the proportion of polyploid cells (≥4N). ·Compared with the HC group, the plasma adiponectin level of the ITP patients significantly increased (P=0.000), and there was no significant correlation between the adiponectin level and the BMI of the patients (P=0.621). RT-qPCR and Western blotting showed that ADIPOR1 and ADIPOR2 were expressed in K562, MEG-01 and Dami cell lines. The proportion of CD41+ cells in the K562 cell line co-cultured with 10 μmol/L or 20 μmol/L AdipoRon and PMA for 72 h were significantly lower than those in the PMA+DMSO group (P=0.000). The CD41-MFI (P=0.047) and the proportion of polyploid cells (P=0.003) in the MEG-01 cell line co-cultured with 20 μmol/L AdipoRon and PMA for 72 h were significantly higher than those in the PMA+DMSO group; however, in the Dami cell line, no significant increases of both were found after the treatment of 20 μmol/L AdipoRon and PMA. ·The adiponectin level is elevated in the peripheral blood plasma of ITP patients. Human myeloid cell line K562, and megakaryocyte cell lines MEG-01 and Dami all express adiponectin receptors ADIPOR1 and ADIPOR2. Adiponectin receptor agonists can inhibit megakaryocyte differentiation in vitro, but its effect on megakaryocyte maturation is uncertain. Figures and Tables | References | Related Articles | Metrics

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Development of pH-responsive fluorescent probe for tumor microenvironment imaging WANG Yuxin, SUN Ruiqi, LIU Jianhua, HE Weina 2022, 42 (7):  875-884.  doi: 10.3969/j.issn.1674-8115.2022.07.005 Abstract ( 444 )   HTML ( 22 )   PDF (4327KB) ( 406 )   ·To synthesize a water-soluble pH-responsive near-infrared cyanine-based fluorescent probe, evaluate its optical properties, and conduct in vivo imaging analysis of the quasi-tumor microenvironment. ·The water-soluble pH-responsive near-infrared fluorescent probe R2S was synthesized via two classic chemical transformation steps and the liposoluble probe R2Z was synthesized in the similar way. 1H nuclear magnetic resonance spectroscopy, mass spectroscopy and high performance liquid chromatography were used to verify the structure and the purity of the probes synthesized. Ultraviolet-visible and photoluminescence spectroscopy were used to evaluate its pH responsiveness, response reversibility and stability. Then, cell permeability of the probe was measured by cell imaging. The cytotoxicity of the probe was evaluated with HCT-116 cells and HeLa cells, and in vivo imaging experiments were performed by using healthy BALB/c female mice. Finally, imaging experiments of quasi-tumor-acidic-microenvironment were carried out by separately injecting PBS solution with pH 6.50 and pH 7.40 into the back of the mice, and the fluorescence intensity of R2S at pH 6.50 and pH 7.40 was compared. ·The water-soluble pH-responsive near-infrared fluorescent probe R2S and its liposoluble analog R2Z were synthesized successfully. As the acidity of the probe solution increased (from pH 11.10 to pH 3.47), the maximum absorption wavelength of R2S shifted from 642 nm to 774 nm, while the maximum emission wavelength shifted from 794 nm to 808 nm, with a pKa value of 6.88. Compared to the liposoluble probe R2Z, R2S showed a larger Stokes shift, higher stability without affecting the cell membrane permeability. At the high administration concentration of 100 μmol/L, R2S did not exhibit the inhibition of proliferation, while for R2Z at the concentration of 12.5 μmol/L, the relative cell viability already descended to below 80%, indicating R2S had higher biosecurity than R2Z. Imaging of R2S in low pH tissue (simulating tumor tissue) and normal pH tissue exhibited a high degree of discrimination. ·The water-soluble pH-responsive near-infrared cyanine fluorescent probe R2S exhibits sensitive and stable responses to pH changes, and the optimal response range is consistent with the pH of the tumor acidic microenvironment, illustrating a strong potential for its in vivo tumor imaging applications. Figures and Tables | References | Related Articles | Metrics

Clinical research

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Clinical study of low dose febuxostat on improving renal function in elderly patients with asymptomatic hyperuricemia LAI Xiuxiu, ZHU Qingyan, TAN Jiaqi, YANG Ling, ZHU Yan, ZHOU Gongmin 2022, 42 (7):  885-892.  doi: 10.3969/j.issn.1674-8115.2022.07.006 Abstract ( 328 )   HTML ( 26 )   PDF (1527KB) ( 146 )   ·To investigate the efficacy, safety and effect on renal function of low-dose febuxostat in the treatment of elderly patients with chronic kidney disease (CKD) complicated with asymptomatic hyperuricemia (HUA). ·In this prospective cohort study, a total of 102 elderly patients who were hospitalized in the Wangjiangshan Branch, Zhejiang Province People's Hospital from February 2021 to July 2021 and met the enrollment conditions were selected. They were divided into low dose group (20 mg/d), normal dose group (40 mg/d) and control group (without urate-lowering drug, mainly lifestyle intervention). The total course of treatment was 3 months. The control rates of serum uric acid (SUA) were calculated in each group monthly. The changes of SUA, serum creatinine (Scr), creatinine clearance rate (Ccr) and estimated glomerular filtration rate (eGFR) were collected and calculated at baseline and after 1 and 3 months of treatment. Meanwhile, eGFR1 was calculated by the simplified diet in renal disease (MDRD) equation, and eGFR2 was calculated by the chronic kidney disease epidemiology collaboration (CKD-EPI) equation. At the same time, cardiovascular and cerebrovascular events, gout attack, allergy and other adverse events caused by febuxostat were recorded. χ2 test was used to compare the differences in gender composition, basic disease and drug use among the three groups. One-way ANOVA was used to compare the differences in SUA, Scr, Ccr, eGFR1 and eGFR2 among the three groups. ·After 1, 2 and 3 months of treatment, the control rates of SUA in the low dose group and normal dose group were 93.8%, 93.8%, 90.6% and 93.8%, 93.8%, 96.9%, respectively. The SUA levels in the above two groups were significantly decreased, compared with the control group (all P=0.000). Ccr, eGFR1, eGFR2 in the low dose group were higher than those at baseline (P=0.006, P=0.013, P=0.015) and those in the control group (P=0.019, P=0.020, P=0.021). There were no significant differences in the decrease of SUA between the normal dose group and the low dose group, but they were significantly greater than those in the control group (all P=0.000). The increase of Ccr, eGFR1 and eGFR2 in the low dose group was significantly higher than that in the control group (P=0.004, P=0.002, P=0.003), but the difference was not statistically significant compared with the normal dose group. Severe adverse events specific to febuxostat were not observed. ·Low dose and normal dose of febuxostat both have a similar urate-lowering efficacy in elderly patients with CKD complicated with asymptomatic HUA, getting a high control rate of SUA and improving renal function without serious adverse events. It is suggested a low dose of febuxostat at the beginning of the clinical intervention of asymptomatic HUA. Figures and Tables | References | Related Articles | Metrics

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Clinical study of preventive extraction of impacted mandibular third molar germ LI Chenlin, LI Yan, XU Guangzhou 2022, 42 (7):  893-897.  doi: 10.3969/j.issn.1674-8115.2022.07.007 Abstract ( 363 )   HTML ( 25 )   PDF (1298KB) ( 110 )   ·To study the short-term and long-term clinical effect of preventive extraction of mandibular third molar (M3M) germ with impaction tendency. ·From September 2018 to April 2020, 21 cases of M3M germs with impaction tendency were included in the experimental group at the Department of Oral Surgery, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine. At the same time, 21 adult M3Ms which were completely impacted were included as the control group. M3Ms were extracted by ultrasonic osteotome and turbine drill. The patients were followed up 1 d, 3 d, 7 d, 3 months and 6 months after surgery. The surgery time, bone defect depth (BDD), probing depth (PD), plaque index (PLI), gingival index (GI), pain degree (visual analog score, VAS), degree of swelling, mouth opening restriction degree, and adverse reactions were compared between the two groups. IBM SPSS statistics 25 software was used for statistical analysis, and the quantitative data were expressed in x±s. The independent sample t-test or Mann-Whitney U test was used for the comparison between the two groups in primary and secondary outcomes. Kruskal-Wallis H test was used to analyze the intragroup changes of the primary outcomes, and Mann-Whitney U test was used for further pairwise comparison within the group. Statistical significance was accepted at a value of P<0.05. ·The surgery time of the experimental group was (7.62±1.50) min and that of the control group was (10.76±2.21) min. There was a significant difference in the surgery time between two groups (P=0.000). The BDD of the experimental group was (0.65±0.33) mm and that of the control group was (2.49±1.37) mm at 6 months after surgery. There were significant differences in BDD, PD, PLI and GI between the two groups at 6 months after surgery (P=0.000, P=0.012, P=0.011, P=0.042). The BDD of the two groups at 6 months after surgery was significantly higher than that at baseline, and the difference was statistically significant (P=0.009, P=0.000). There was a significant difference in VAS between the two groups at 3 d after surgery (P=0.044). At each follow-up period after surgery, there was a significant difference in the degree of swelling between the two groups (P=0.005, P=0.031, P=0.019). In the control group, 1 case of inferior alveolar nerve injury occurred, and the other patients did not have adverse reactions such as bleeding, nerve injury, dry socket disease, etc. ·Preventive extraction of M3M germ with impaction tendency can shorten the surgery time, and reduce postoperative reaction, which can obtain better long-term clinical effect compared with adult impacted M3M extraction. Figures and Tables | References | Related Articles | Metrics

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Predictive value of fibroblast growth factor 19 subgroup in early pregnancy on gestational diabetes mellitus GAO Fei, LU Yu, DONG Shuqin, YANG Liu, WU Shaohua, WEI Jing 2022, 42 (7):  898-903.  doi: 10.3969/j.issn.1674-8115.2022.07.008 Abstract ( 276 )   HTML ( 23 )   PDF (1369KB) ( 104 )   ·To investigate the relationship between serum levels of fibroblast growth factor 19 subgroup (FGF19, 21 and 23) and the risk of gestational diabetes mellitus (GDM) in early pregnancy. ·The levels of fasting blood glucose (FPG), blood lipid, FGF19, FGF21 and FGF23 of women in the early pregnancy (8-12 weeks) who underwent obstetric examination in Taizhou People's Hospital from October 2019 to October 2020 were collected. According to the results of 75 g oral glucose tolerance test at 24-28 weeks of pregnancy, they were divided into GDM group and NGT group. The levels of FGF19, FGF21 and FGF23 were compared between the two groups, and their relationship with GDM were analyzed. ·A total of 180 pregnant women were followed up, including 27 cases in the GDM group and 153 cases in the NGT group. Compared with the NGT group, FGF21 levels in the GDM group were significantly higher, while the levels of FGF19 and FGF23 were significantly lower (P<0.05, respectively). Correlation analysis showed that FPG was negatively correlated with FGF19 (r=-0.283, P<0.01), and positively correlated with LogFGF21 (r=0.160, P<0.05), and had no significant correlation with LogFGF23 (P>0.05). Single factor regression analysis showed that FGF19, FGF21, FGF23 and FPG were influencing factors of GDM (P<0.05, respectively). Multivariate Logistic regression analysis showed that FGF21 and FPG were risk factors for GDM (OR=1.196, P=0.000; OR=2.280, P=0.010). The areas under the ROC curve of FGF21, FPG and their combined prediction of GDM were 0.707, 0.646 and 0.847. ·The levels of FGF21 and FPG increase, while the levels of FGF19 and FGF23 decrease in GDM pregnancies in early pregnancy. FGF21 and FPG are risk factors of GDM. The FGF21- and FPG- combined prediction of GDM has greater diagnostic efficiency. Figures and Tables | References | Related Articles | Metrics

Public health

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Effect of combination of medical care and nursing on the quality of life and mental state of elderly patients with chronic kidney disease in Shanghai suburbs WANG Yakun, XU Jiarui, WU Qianqian, ZHANG Xiaohua, ZHU Yingchun, BAI Shoujun 2022, 42 (7):  904-910.  doi: 10.3969/j.issn.1674-8115.2022.07.009 Abstract ( 338 )   HTML ( 17 )   PDF (1358KB) ( 146 )   ·To explore the effect of the combination of medical care and nursing on the quality of life and the anxiety/depression symptom of the elderly patients with chronic kidney disease (CKD) in Shanghai surburbs. ·The elderly patients with CKD living in two nursing homes in Qingpu District, Shanghai were selected. The anxiety symptom was assessed by Generalized Anxiety Disorder Scale-7 (GAD-7), the depression symptom was assessed by The Geriatric Depression Scale (GDS), and the quality of life was assessed by The MOS 36-item Short Form Health Survey (SF-36). According to whether they had anxiety/depression symptom, they were divided into one group with mental symptoms (group A) and the other group without mental symptoms (group B). Then, the patients in group A were randomly divided into medical care and nursing-combined intervention group (group A1) and conventional group (group A2). Except group A1, group A2 and group B both adopted the traditional chronic disease management mode. The correlations of the scores of SF-36 in 8 dimensions with the GAD-7 score and the GDS score were analyzed. Multivariate Logistic regression analysis was used to analyze the independent risk factors of life quality of elderly patients with CKD. The mental symptoms and the quality of life before and after the 6-month interventions were compared to evaluate the combination of medical care and nursing. ·A total of 80 elderly patients with CKD were enrolled, including 54 cases (67.5%) in group A and 26 cases (32.5%) in group B, whose incidences of anxiety symptom and depression symptom were 37.5% and 30.0%, respectively. Group A included 27 cases in group A1 and 27 cases in group A2. The GAD-7 score and the GDS score were negatively correlated with the scores of 6 dimensions of quality of life (physiological function, role-physical, general health, vitality, role-emotional, and mental health), respectively (P<0.05). Logistic regression analysis showed that anxiety/depression symptom was an independent risk factor of the quality of life of elderly CKD patients. After the combined intervention of medical care and nursing for 6 months, the scores of GAD-7 and GDS in group A1 were significantly lower than those before intervention (P<0.05), while there was no statistical difference in the scores before and after the intervention in group A2. Before intervention, the scores of 8 dimensions of quality of life in group A1 and A2 were significantly lower than those in group B (P<0.05), and there was no significant difference between A1 and A2 groups. After 6 months of intervention, the scores of 6 dimensions in group A1 were significantly higher than those in group A2 and those in group A1 before the intervention (P<0.05), and there were no obvious changes in role-physical and social functioning. ·The anxiety/depression symptom of elderly patients with CKD in Shanghai surburbs can reduce their quality of life; the combination of medical care and nursing can improve the anxiety/depression symptom of these patients and their quality of life as well. Figures and Tables | References | Related Articles | Metrics

Techniques and methods

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Effects of different expression matrices on screening differential lncRNAs WEI Hao, QIU Jiajun, YAN Jingbin 2022, 42 (7):  911-918.  doi: 10.3969/j.issn.1674-8115.2022.07.010 Abstract ( 230 )   HTML ( 20 )   PDF (2437KB) ( 129 )   ·To compare the effects of two methods for differential analysis of long non-coding RNA (lncRNA) expression levels on screening differential lncRNAs based on whole transcriptome sequencing data. ·Two sets of whole transcriptome sequencing datasets were downloaded from the NCBI_GEO database with a total of 10 samples. Group A consisted of universal human reference RNA samples, and Group B consisted of human brain reference RNA samples. Each sample contained a series of synthetic RNA (spike-in RNA) at known concentrations from the External RNA Control Consortium (ERCC). The processed sequencing data were counted by using the annotated reference genomes of mRNA, lncRNA, and total RNA, respectively, to obtain the corresponding three expression matrices containing the annotation information of spike-in RNA. Under the condition of P<0.05, according to the real concentration of spike-in RNA in different groups, the false positive rate and false negative rate of differential expression analysis results were judged. The R language software packages DESeq2 and edgeR were used to perform differential expression analysis between groups for all expression matrices, and the receiver operating characteristic (ROC) curve of spike-in RNA was used to show the specificity and sensitivity of differential expression analysis of different expression matrices. Our study mainly focused on the differences between the total RNA expression matrix and the lncRNA expression matrix. Differentially expressed lncRNA analysis was then performed on the total RNA expression matrix and lncRNA expression matrix within groups, and the P value distribution was calculated to compare the false positive rate of different expression matrices. ·Under the condition of P<0.05, the false positive rate and false negative rate of spike-in RNA between group A and B were 0.52 and 0.14 when analyzed with the total RNA expression matrix, and when analyzed with the lncRNA expression matrix, it was 0.30 and 0.17, which indicated that the false positive rate using the lncRNA expression matrix differential analysis was higher. The area under the curve (AUC) of spike-in RNA in expression matrices analyzed by different R packages was generally consistent: AUC (total RNA)≈AUC (mRNA)<AUC (lncRNA), which indicated that the screening effect of lncRNA expression matrix was better than that of total RNA. The intra-group lncRNA differential expression analysis results showed that, under the condition of P<0.05, there were 9 and 7 different expressed lncRNAs in the lncRNA expression matrix and total RNA expression matrix in group A, and 15 and 17 in group B, respectively. The numbers were not significantly different between expression matrices. ·In the differential expression analysis of known lncRNAs in whole transcriptome sequencing data, the specificity and sensitivity of the lncRNA expression matrix analysis are better than that of total RNA. Figures and Tables | References | Related Articles | Metrics

Review

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Research progress of clinical application of ASPECT score in acute ischemic stroke WEI Xuemin, GAO Chengjin 2022, 42 (7):  919-924.  doi: 10.3969/j.issn.1674-8115.2022.07.011 Abstract ( 646 )   HTML ( 26 )   PDF (1187KB) ( 456 )   Acute ischemic stroke (AIS) is characterized by a high rate of disability and mortality, and rapid and accurate diagnosis and appropriate treatment are the keys. The severity of AIS patients depends on the location of early ischemic changes (EIC) in the brain tissue and the size of the lesion, so it is necessary to evaluate the patients' conditions by combining with imaging. The Alberta stroke program early CT score (ASPECT score) is a 10-point imaging score system that evaluates the EIC in the middle cerebral artery supplying region of the stroke patients in an accurate and simple way by quantifying the severity in terms of the imaging to help clinicians make medical decisions. ASPECT score is not only widely used to predict the prognosis of stroke patients, but also to identify the population benefiting from the endovascular therapy and evaluate the risk of endovascular therapy for patients. ASPECT score combined with multimodal imaging has also been used to predict ischemic core volumes and the prognosis. In recent years, with the development of artificial intelligence, automatic ASPECT scoring methods based on machine learning have emerged. This paper reviews ASPECT scoring methods, its value in the treatment and the assessment of prognosis in AIS, multimodal ASPECT scoring, and the application of automated ASPECT scoring combined with artificial intelligence. References | Related Articles | Metrics

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Role of SUMOylation in spermatogenesis LUAN Jiayan, LI Peng, HAN Bangmin 2022, 42 (7):  925-930.  doi: 10.3969/j.issn.1674-8115.2022.07.012 Abstract ( 417 )   HTML ( 22 )   PDF (1445KB) ( 144 )   SUMOylation is a reversible post-translational modification of proteins, which is involved in many important biological processes, such as transcriptional regulation, signal transduction, and DNA damage repair. It is found that the overall expressions and localizations of SUMOylation change dynamically in spermatogenesis, suggesting that it may be involved in key events in the process. Spermatogenesis is a series of processes regulated by complex biological crosstalk. In the seminiferous tubules, spermatogenic cells undergo three stages, that is, the proliferation and differentiation of spermatogonial stem cells, the meiosis of spermatocytes and spermiogenesis, and eventually form highly specialized spermatozoa. In this process, the effects and mechanisms of SUMOylation in spermatogenesis have not been clear. Therefore, this review summarizes the effect of the SUMOylation enzyme system, SUMO family and SUMO-specific proteases family in spermatogenesis. Figures and Tables | References | Related Articles | Metrics

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Progress in pathogenesis and clinical treatment of sarcoidosis ZHANG Lincheng, ZHONG Hua 2022, 42 (7):  931-938.  doi: 10.3969/j.issn.1674-8115.2022.07.013 Abstract ( 289 )   HTML ( 23 )   PDF (1303KB) ( 164 )   Sarcoidosis is a systemic disease which could affect many crucial organs and eventually cause irreversible functional damage to these organs. However, the pathogenesis of sarcoidosis is still not clear. Current studies suggest that sarcoidosis is caused by multiple factors. Individuals with certain genetic variations are highly susceptible to certain types of sarcoidosis. Furthermore, exposure to extrinsic antigenic factors such as pathogens or inorganic factors and intolerance of self-antigens also play a crucial role in the pathogenesis of sarcoidosis. Subsequent to antigenic stimulation, abnormality in pathways of pattern recognition receptor, phagosome or autophagy stimulates the activity of CD4+ T cells which release inflammatory cytokines such as interleukin (IL)-12, interferon-γ, tumor necrosis factor-α, IL-17, and IL-23, thus fostering the proliferation of Th1 and Th17 subpopulation and inducing the fusion of macrophages to form granulomas. Here we aim to review mechanistic studies in the past decade and list current therapeutic methods to provide some insights into the etiology, pathogenesis and management of sarcoidosis. References | Related Articles | Metrics

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Role of gut microbiota in hepatocellular carcinoma: cancer occurrence, progresses and treatments LU Yu, WANG Hao, BA Qian 2022, 42 (7):  939-944.  doi: 10.3969/j.issn.1674-8115.2022.07.014 Abstract ( 358 )   HTML ( 19 )   PDF (1285KB) ( 168 )   The gut microbiota, as one of the microfloras existing in human body, is accounted for approximately 80% in human microbiome. Gut microbiota regulates digestive and immune functions through participating in physiological processes such as metabolism and immune homeostasis, meanwhile impacts the occurrence of cancer and controls the body response to cancer treatment drugs. Hepatocellular carcinoma (HCC) is a typical type of inflammatory cancer, the development of which follows the process of hepatitis, liver fibrosis, liver cirrhosis and HCC. The bidirectional communication system composed of gut microbiota, portal vein system and bile duct system, is usually called the enteric-liver axis. More and more research evidences show that gut microbiota can participate in the occurrence and development of HCC via the interaction cycle with the enteric-liver axis. In view of the specific transformation of gut microbiota in the early stage of HCC, it is usually regarded as a new target for the early diagnosis of HCC. Immunotherapy is an emerging treatment for advanced HCC. It is reported that regulating gut microbiota through probiotics, rational application of antibiotics, fecal transplantation and other methods can significantly improve the anti-tumor immune response of host. In the non-immunotherapy process of HCC chemotherapy, traditional Chinese medicine-assisted therapy and dietary pattern, remodeling gut microbiota homeostasis has potential value in postponing tumor progression, improving the prognosis of HCC treatment and maintaining body health. Since gut microbiota plays a crucial role in the occurrence, development and treatment of HCC, this review summarizes the studies of gut microbiota in the process of hepatitis, liver fibrosis, cirrhosis and HCC, and discusses its great potential as a target for the prevention, diagnosis and treatment of HCC. References | Related Articles | Metrics

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Advances in the correlation between cytokine signal transduction inhibitors and rheumatoid arthritis LEI Haitao, TIAN Xuemei, JIN Fangquan 2022, 42 (7):  945-951.  doi: 10.3969/j.issn.1674-8115.2022.07.015 Abstract ( 358 )   HTML ( 23 )   PDF (1272KB) ( 183 )   Rheumatoid arthritis (RA) is a chronic symmetrical disease with synovial inflammation, bone destruction and pannus formation. Inflammatory mediators are closely related to the pathogenesis of RA, and the severity of RA is significantly affected by the promotion and demotion of inflammatory mediator levels. In this review, we have reviewed the studies about the correlation between cytokine signaling inhibitory factors and RA in recent years. We have found that cytokine signaling inhibitory factors, as endogenous negative regulators, are involved in intra- and extra-cellular signal transduction, mediating T cell survival and differentiation, inducing transcription and activation of inflammatory factors, and playing a regulatory role in the activation of various immune responses invivovia blocking the phosphorylation of signal transducer and activator of transcription (STAT) and inhibiting the activity of Janus kinase (JAK). Therefore, as an anti-inflammatory mediator protein, cytokine signaling inhibitory factors are involved in inflammatory responses via various pathways, closely related to the occurrence and development of RA, and are expected to be a biomarker for the diagnosis and prognosis for RA. Meanwhile, we predict that the study of apoptosis, immune regulation, cartilage metabolism and autophagy related to cytokine signaling inhibitors may become a research hotspot for RA in the future. Further in-depth research on the correlation and mechanism of cytokine signaling inhibitory factors and RA may widen our vision for the treatment of RA. References | Related Articles | Metrics

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Progress in mechanism of transcranial direct current stimulation WANG Yang, CHENG Jiayue, WANG Zhen 2022, 42 (7):  952-957.  doi: 10.3969/j.issn.1674-8115.2022.07.016 Abstract ( 573 )   HTML ( 29 )   PDF (1199KB) ( 275 )   Transcranial direct current stimulation (tDCS), as a non-invasive brain stimulation technique,is widely used in improving brain function and behavior. Growing evidence shows its broad therapeutic promise in neuropsychiatric disorders. However, there is a lot of uncertainty about how this approach works. This review focuses on the current research on the mechanism of tDCS. The immediate effect of tDCS is mainly reflected in the change of membrane potential of neurons, while the aftereffect of tDCS can cause dynamic changes in Ca2+ concentration, regulate the release and distribution of various neurotransmitters, directly or indirectly alter synaptic plasticity, and play an important role in the regulation of synaptic metaplasticity. In addition to regional effects, tDCS results in widespread network-level changes across the brain. This review probes into the predicament of current research and puts forward some unsolved problems. References | Related Articles | Metrics

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Analysis of early clinical symptoms and surgical intervention of hemorrhagic cystitis after hematopoietic stem cell transplantation in children LIU Pengtao, ZHANG Zhiyuan, BAI Kaiping, XING Xiaoyu, ZOU Xiangyu, SUN Jie 2022, 42 (7):  958-963.  doi: 10.3969/j.issn.1674-8115.2022.07.017 Abstract ( 327 )   HTML ( 16 )   PDF (1364KB) ( 92 )   ·To analyze the characteristics of early clinical symptoms of hemorrhagic cystitis (HC) after allogeneic hematopoietic stem cell transplantation (HSCT) and the relationship between the symptoms and disease progression. ·The clinical data of 77 children with HC after HSCT, treated in Shanghai Children's Medical Center, Shanghai Jiao Tong University School of Medicine from June 2013 to December 2020, were retrospectively analyzed. Each child was prophylactically applied with mesna in the early stage, and hydration, alkalization, diuretics, anti-infection agents, antispasmodic and analgesic drugs were applied during the course of disease. Moreover, indwelling catheterization and bladder perfusion or cystoscopic electrical coagulation were performed for refractory HC children; Open cystostomy combined with postoperative bladder perfusion was performed when the cystoscope was difficult to perform. HC was graded according to Droller's standard, and the characteristics of urinary irritation symptoms and hematuria symptoms, the onset time of each symptom, the time of progression to severe HC (grade Ⅲ?Ⅳ), the duration of disease course, treatment methods and therapeutic effects were recorded. ·Among the 77 cases, 49 (63.6%) were male and 28 (36.4%) were female. The median age was 7 (5, 11) years old. Sixty-five cases (84.4%) had virus infection during the course of disease, such as BK polyomavirus (BKV), cytomegalovirus (CMV) and Epstein-Barr virus (EBV). Forty-five cases (58.4%) had urinary irritation symptoms as the first symptom of urinary tract, and 32 cases (41.6%) had hematuria. Children who had urinary irritation symptoms as the first symptom eventually developed hematuria symptoms of varying degrees after hydration, alkalization, analgesic drugs or other routine treatments, with a median interval of 2 (1, 5) d, including 3 cases of grade Ⅰ (6.7%), 16 of grade Ⅱ (35.5%), 23 of grade Ⅲ (51.1%), and 3 of grade Ⅳ (6.7%). There were 3 cases of surgical treatment: 1 case of grade Ⅲ was completely relieved after cystoscopic hemostatic treatment, 1 case of grade Ⅳ was partially relieved after open cystostomy combined with postoperative bladder perfusion, and 1 case of grade Ⅳ died for multiple organ failure after cystoscopic hemostatic treatment. ·More than half of the children with HC after HSCT start with urinary irritation symptoms, and more than half of the children with severe HC also start with it. Such children need more active intervention measures in the early stage. Figures and Tables | References | Related Articles | Metrics

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Two cases of leukoencephalopathy with vanishing white matter caused by new mutation of EIF2B gene and literature review LIU Taotao, WU Jingying, LIU Xiaoli, ZHANG Mei, CAO Li 2022, 42 (7):  964-970.  doi: 10.3969/j.issn.1674-8115.2022.07.018 Abstract ( 313 )   HTML ( 13 )   PDF (3620KB) ( 174 )   Two cases of leukoencephalopathy with vanishing white matter (VWM) were reported. Patient 1 was a two years and three months old girl whose main clinical manifestation was rapidly progressive motor disturbance following recurrent infections. She developed epilepsy after 6 months. After that, she was unable to walk independently and had difficulty in swallowing. Genetic testing revealed that there were compound heterozygous mutations in the EIF2B4 gene of the proband, including the c.594C>G (p.I98M) in exon 7 inherited from father and the c.1177T>A (p.Y393N) in exon 11 inherited from mother that both had not been reported previously. Cranial MRI showed diffuse symmetrical white matter lesions with prominent white matter thinning. Patient 2 was a 41-year-old female, the second adult VWM case reported in China, mainly manifesting as progressive weakness in both lower limbs and memory loss. Genetic testing revealed that there were compound heterozygous mutations in the EIF2B3 gene of the proband, including the c.130G>A (p.G44K) in exon 2 and the c.934C>G (p.R312G) in exon 8, in which the latter mutation type was reported for the first time. Cranial MRI showed white matter atrophy and ventriculomegaly. Age was a clinical predictor of the severity of VWM, and specifically earlier onset could be associated with more severe disability and higher mortality. Stress was a trigger for the disease and could contribute to neurological deterioration. At present, there is no effective therapy method for this disease. This article reports the two cases, aiming to improve clinicians' understanding of the disease and their ability of early diagnosis, so as to delay the progress of the disease and prolong the survival of the patients. Figures and Tables | References | Related Articles | Metrics