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    Basic research
    Effect of Leptospira interrogans 56606v on apoptosis of murine neutrophil
    DU Lin, HE Ping
    2022, 42 (11):  1517-1523. 
    doi: 10.3969/j.issn.1674-8115.2022.11.001

    Abstract ( 177 )   HTML ( 28 )   PDF (3302KB) ( 68 )  

    Objective ·To investigate the effect of Leptospira interrogans 56606v on apoptosis of murine neutrophils and related mechanisms. Methods ·Murine neutrophils were infected with Leptospira interrogans 56606v at indicated multiplicity of infection (MOI was 100, 300 and 500 respectively) and time points (0.5,6,12 and 24 h), with uninfected neutrophils as control. Cells treated with an equal amount of cell culture medium containing 4 μmol/L astrosporine (STS) were used as positive control for apoptosis. The apoptosis was analyzed by using flow cytometry. The cleavage of caspase-3 and activation of nuclear factor-κB (NF-κB) pathway were detected in murine neutrophils infected with Leptospira interrogans 56606v by Western blotting, and the inhibitory effect of NF-κB inhibitor BAY 11-7082 on NF-κB pathway was also verified by Western blotting. The role of NF-κB pathway in the inhibition of neutrophil apoptosis induced by Leptospira interrogans 56606v was further explored by adding NF-κB inhibitor BAY 11-7082. Macrophages were co-incubated with Leptospira-infected or-uninfected carboxyfluorescein diacetate succinimidyl ester (CFSE)-labeled neutrophils, and flow cytometry was used to detect the phagocytosis of neutrophils by macrophages to reflect the effect of the neutrophil apoptosis inhibition induced by Leptospirainterrogans 56606v infection on efferocytosis. Results ·The results of flow cytometry showed that the apoptosis rate of neutrophils in Leptospira infection group was decreased compared with the uninfected control group. The inhibition of Leptospirainterrogans 56606v on neutrophil apoptosis was enhanced with the increase of MOI. Compared with the uninfected control group, the difference was statistically significant under different MOIS (P=0.000). The apoptosis rate was lower than that of the corresponding uninfected control group when Leptospirainterrogans 56606v with MOI=100 infected neutrophils for 0.5, 6 and 12 h (P<0.05), and the apoptosis of neutrophils was enhanced after treatment with STS. The cleavage of caspase-3 was significantly reduced after infection with Leptospira interrogans 56606v and the phosphorylation of NF-κB was significantly increased after infection demonstrated by Western blotting. The activation of NF-κB pathway in both post-infection and non-infection conditions was significantly inhibited by BAY 11-7082. The apoptosis rate of murine neutrophils infected with or without Leptospira interrogans 56606v was significantly increased by BAY 11-7082 (P=0.000). The neutrophils infected with Leptospira interrogans 56606v were significantly less phagocytosed by macrophages than those uninfected (P=0.027). Conclusion ·Leptospira interrogans 56606v inhibits the apoptosis of murine neutrophils by activating the NF-κB pathway, which further reduces efferocytosis of macrophages on neutrophils.

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    Genomic regulatory network of human olfactory neuroepithelial cells infected with novel coronavirus (SARS-COV-2)
    XIE Xinyi, YANG Yumeng, WANG Shaowei, SUN Na, SHI Chuandao, LIU Qiling, ZHANG Rongqiang, LI Junjie
    2022, 42 (11):  1524-1533. 
    doi: 10.3969/j.issn.1674-8115.2022.11.002

    Abstract ( 165 )   HTML ( 17 )   PDF (4813KB) ( 55 )  

    Objective To explore the genomic changes of human olfactory neuroepithelial cells after the novel coronavirus (SARS-COV-2) infecting the human body, and establish a protein-protein interaction (PPI) network of differentially expressed genes (DEGs), in order to understand the impact of SARS-COV-2 infection on human olfactory neuroepithelial cells, and provide reference for the prevention and treatment of new coronavirus pneumonia. Methods The public dataset GSE151973 was analyzed by NetworkAnalyst. DEGs were selected by conducting Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) signal pathway analysis. PPI network, DEGs-microRNA regulatory network, transcription factor-DEGs regulatory network, environmental chemicals-DEGs regulatory network, and drug-DEGs regulatory network were created and visualized by using Cytoscape 3.7.2. Results After SAR-COV-2 invading human olfactory neuroepithelial cells, part of the gene expression profile was significantly up-regulated or down-regulated. A total of 568 DEGs were found, including 550 up-regulated genes (96.8%) and 18 down-regulated genes (3.2%). DEGs were mainly involved in biological processes such as endothelial development and angiogenesis of the olfactory epithelium, and the expression of molecular functions such as the binding of the N-terminal myristylation domain. PPI network suggested that RTP1 and RTP2 were core proteins. MAZ was the most influential transcription factor. Hsa-mir-26b-5p had the most obvious interaction with DEGs regulation. Environmental chemical valproic acid and drug ethanol had the most influence on the regulation of DEG. Conclusion The gene expression of olfactory neuroepithelial cells is significantly up-regulated or down-regulated after infection with SAR-COV-2. SARS-CoV-2 may inhibit the proliferation and differentiation of muscle satellite cells by inhibiting the function of PAX7. RTP1 and RTP2 may resist SARS-CoV-2 by promoting the ability of olfactory receptors to coat the membrane and enhancing the ability of olfactory receptors to respond to odorant ligands. MAZ may regulate DEGs by affecting cell growth and proliferation. Micro RNA, environmental chemicals and drugs also play an important role in the anti-SAR-COV-2 infection process of human olfactory neuroepithelial cells.

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    lncRNA NEAT1 affects the proliferation, invasion and migration of ox-LDL-induced human vascular smooth muscle cells through miR-377-3p/Wnt pathway
    YUAN Yongmei, CHENG Xiaodan, SUN Jiaan, CHANG Dongge, HE Yingying, LIU Chang
    2022, 42 (11):  1534-1541. 
    doi: 10.3969/j.issn.1674-8115.2022.11.003

    Abstract ( 171 )   HTML ( 24 )   PDF (4786KB) ( 50 )  

    Objective ·To study the effects of long non-coding RNA (lncRNA) nuclear paraspeckle assembly transcript 1 (NEAT1) on the proliferation, invasion and migration of oxidized low-density lipoprotein (ox-LDL)-induced human vascular smooth muscle cells (HVSMCs) through regulating microRNA-377-3p (miR-377-3p) /Wnt pathway. Methods ·HVSMCs cultured in vitro were treated with 100 μg/mL ox-LDL. HVSMCs were randomly divided into control group, model group, NEAT1 siRNA group, miR-377-3p inhibitor group, transfection+inhibitor (NEAT1 siRNA+miR-377-3p inhibitor) group, and transfection-negative control (NEAT1 siRNA-negative control) group. Except for the control group, the atherosclerosis cell models were induced with 100 μg/mL ox-LDL in other groups. After grouping and transfection, the cell viability of each group was determined by using cell counting kit-8 (CCK-8) experiment; the apoptosis rate of each group was measured by flow cytometry; the migration and invasion of cells in each group were evaluated by the Transwell experiment; the expression of cell proliferation markers such as proliferating cell nuclear antigen (PCNA), and epithelial-mesenchymal transition (EMT) markers, such as E-cadherin and vimentin in each group, was determined by immunoblotting experiment; the expression of NEAT1, miR-377-3p and wingless-related MMTV integration site 5A (WNT5A) mRNA in each group was determined by qRT-PCR experiment. Results ·Compared with the control group, the mRNA expression of NEAT1 in the model group increased, the mRNA expression of miR-377-3p decreased, the apoptosis rate decreased, the cell viability, and the number of migrating and invasive cells increased, the protein expression of PCNA and Vimentin increased, the expression of E-cadherin decreased, and WNT5A mRNA expression increased (all P=0.000). After NEAT1 was interfered, the apoptosis rate increased, the cell viability, and the number of migrating and invasive cells decreased, the protein expression of PCNA and Vimentin decreased, the expression of E-cadherin increased, and WNT5A mRNA expression decreased (all P=0.000); after miR-377-3p was inhibited, all the above indicators showed opposite expression trends (P<0.05). In addition, inhibition of miR-377-3p reversed the regulation of NEAT1 interference on cell proliferation, invasion, migration and apoptosis (all P=0.000). Conclusion ·The down-regulating of NEAT1 expression may increase miR-377-3p expression by competitive binding to miR-377-3p, reduce protein expression associated with the Wnt pathway, inhibit the abnormal proliferation, invasion and migration of HVSMCs induced by ox-LDL, and induce apoptosis.

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    Anticancer effect of drug-loaded DNA nanoflowers on mouse lung cancer cells
    LIAO Hongjian, CAO Yuchao, DU Yonghong
    2022, 42 (11):  1542-1549. 
    doi: 10.3969/j.issn.1674-8115.2022.11.004

    Abstract ( 178 )   HTML ( 25 )   PDF (4046KB) ( 55 )  

    Objective ·To investigate the anti-cancer effects of DNA nanoflowers (DF) loaded with horseradish peroxidase (HRP) and doxorubicin (DOX) on mice Lewis lung carcinoma (LLC) cells. Methods ·The DF loaded with HRP (HRP-DF) was synthesized by rolling circle amplification (RCA) reaction and DOX was loaded to prepare DOX/HRP-DF, the physical properties of which were detected by scanning electron microscopy and laser particle sizer. In vitro drug release capacity of DOX/HRP-DF was tested with acid buffer solution. The safety of DOX/HRP-DF was evaluated by CCK-8 assay to detect the cytotoxicity of 16HBE cells treated with DOX/HRP-DF and free DOX at different time points. Under anoxic conditions, the oxygen production capacity of DOX/HRP-DF in LLC cells was detected by using the oxygen-sensitive probe, and the effects of different dosages of free DOX, DOX-DF and DOX/HRP-DF on the proliferation capacity of LLC cells were examined by CCK-8 assay. The effects of DOX/HRP-DF on the migration and invasion of LLC cells were detected by scratching assay and Transwell assay in the experimental group treated with different dosages of DOX/HRP-DF and the control group treated with Phosphate Buffered Saline (PBS). Results ·DOX/HRP-DF were successfully prepared and observed as flower-like structures under scanning electron microscopy with uniform size. The average diameter and potential of DOX/HRP-DF were (415.90±9.32) nm and (-17.43±1.20) mV, respectively. DOX/HRP-DF had a DOX loading of (38.19±0.39)% with good pH-sensitive drug release properties. DOX/HRP-DF could significantly reduce the toxic effect of DOX on human normal cells. The results of Laser confocal microscopy showed that DOX/HRP-DF could effectively alleviate the hypoxic condition of LLC cells. CCK-8 assay results showed that the proliferation rate of LLC cells in the DOX/HRP-DF group was significantly lower in a dose-dependent manner, compared to the free DOX and DOX-DF groups (P<0.05). The results of scratching assay showed that the scratch healing rate of the DOX/HRP-DF group was lower than that of PBS group in a dose-dependent manner (P<0.05). Transwell experiments showed the number of LLC cells passing through Matrigel gel in the DOX/HRP-DF group was reduced in a dose-dependent manner, compared with the PBS group (P<0.05). Conclusion ·The successfully prepared DOX/HRP-DF could effectively deliver drugs, and alleviate hypoxia in the tumor microenvironment, and significantly inhibit the proliferation, migration and invasion ability of LLC cells, which is expected to provide new ideas for lung cancer treatment.

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    Clinical research
    Comparison of the root coverage and esthetic outcomes of 3 different techniques for gingival recession
    SUN Wentao, SUN Mengjun, XIE Yufeng, SHU Rong
    2022, 42 (11):  1550-1556. 
    doi: 10.3969/j.issn.1674-8115.2022.11.005

    Abstract ( 238 )   HTML ( 16 )   PDF (2194KB) ( 55 )  

    Objective ·To evaluate the outcomes of connective tissue graft (CTG) combined with 3 different techniques for gingiva recession (GR) including envelope technique, tunnel technique (TUN) and vestibular incision subperiosteal tunnel access (VISTA), and analyze the differences of root coverage and esthetic outcomes of the 3 techniques. Methods ·A total of 87 patients who visited the Department of Periodontology, Shanghai Ninth People′s Hospital, Shanghai Jiao Tong University School of Medicine from January 2020 to December 2021 with a total of 324 GRs were enrolled in this study. All GRs were treated with one of the 3 techniques. The patients′ periodontal conditions were examined at baseline and 6 months after surgery. The root coverage esthetic score (RES) and mucosal scarring index (MSI) were evaluated by 2 periodontists 6 months after surgery. The differences of keratinized gingiva (KG) and GR at baseline and 6 months after surgery were compared by using t-test. Analysis of variance was used to compare the differences of percentage of root coverage (PRC), RES and MSI of different techniques, the same technique in different regions, and different techniques in each region. Results ·In this study, KG increased by (1.49±1.36) mm, and there was a significant difference between each technique (P=0.002). GR decreased by (2.37±1.37) mm, and there was a significant difference between each technique (P=0.000). The mean PRC was (87.7±27.1)%, which was significantly different between each technique (P=0.003). The percentage of complete root coverage (PCRC) was 74.0%, and there was significant difference among the 3 techniques (P=0.000). There were significant differences in RES in different regions between envelope+CTG and VISTA+CTG (Penvelope=0.003, PVISTA=0.000). There was a significant difference in MSI of different regions in VISTA+CTG (P=0.000). Among the 3 techniques, only PRC had differences in the lower anterior teeth (P=0.011); there was a significant difference in RES between lower anterior teeth and lower posterior teeth (PLA=0.001,PLP=0.034), the RES of lower anterior teeth treated with TUN+CTG was higher, and the RES of lower posterior teeth treated with TUN+CTG and VISTA+CTG was higher; there were significant differences in MSI in each region (PUA=0.011, PUP=0.000, PLA=0.003, PLA=0.001). Conclusion ·All the 3 techniques are capable of reducing GR and widening KG. The root coverage and esthetic outcomes of TUN+CTG are superior to the other 2 techniques if the operator′s experience is not considered.

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    Characteristics of mandible deviation in adolescent patients with temporomandibular joint anterior disc displacement
    WAN Shaonan, LI Peilun, XIE Qianyang, CHEN Jun, QIAN Ziyue, YANG Chi
    2022, 42 (11):  1557-1561. 
    doi: 10.3969/j.issn.1674-8115.2022.11.006

    Abstract ( 179 )   HTML ( 14 )   PDF (1356KB) ( 22 )  

    Objective ·To explore the characteristics of mandibular deviation (MD) in adolescent patients with anterior disc displacement (ADD) and the relationship between temporomandibular joint (TMJ) anterior disc displacement and mandibular deviation. Methods ·Consecutive cases aged 12 to 15 who has been treated in the Department of Oral Surgery of Shanghai Ninth People's Hospital from January 2019 to December 2019 were reviewed. Among them, cases with TMJ symptoms, having been diagnosed by MRI as unilateral or bilateral TMJ ADD, were separately included into unilateral displacement group and bilateral displacement group. Cases without TMJ symptoms, showing a normal disc-condyle relationship with MRI, were included into the control group. The posteroanterior cephalometric radiographs of the three groups were collected respectively before orthodontic treatment, which were measured independently by two surveyors. SPSS 23.0 software package was used for statistical processing, and Shapiro-Wilk was used to test its normality. Pearson chi-square test was used to analyze the distribution of mandibular deviation in the three groups, and the degrees of horizontal deviation of submental point in the three groups were compared. The distribution characteristics of mandibular deviation in cases with disc displacement were observed by comparing the mandibular deviation between juvenile patients with TMJ anterior disc displacement and those with normal disc-condyle relationship. Results ·A total of 1 011 cases of 12-15-year-old adolescent patients in our hospital were collected, including 192 cases of unilateral disc displacement group and 255 cases of bilateral disc displacement group. A total of 564 cases were collected in the control group. Both the control group and the disc displacement group showed a skewed distribution. The mean value of MeH in the control group was 2.1 mm. While 3.6 mm was taken as the critical value to distinguish deviation and non-deviation, the proportion of patients with mandibular deviation in the control group, unilateral displacement group and bilateral displacement group was respectively 25.53%, 27.08% and 19.22%. The incidence rate of mandibular deviation was significantly different between the unilateral displacement group and bilateral displacement group (P=0.049). Among all cases with mandible deviation, 68.16% (167/245) were mild deviation (3.6 mm≤MeH<5.8 mm), 21.22% (52/245) were moderate deviation (5.8 mm≤MeH<8.2 mm), and 10.61% (26/245) were severe deviation (MeH≥8.2 mm). Conclusion ·It is found that about 75% of adolescents have mandibular deviation within 3.6 mm; Patients with unilateral anterior disc displacement have a greater probability of deviation than the general population.

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    Clinical and pathological characteristics as well as prognostic analysis of breast cancer patients with brain metastases
    SU Juncheng, WANG Yuzheng, TANG Lei, XU Yingchun, ZHANG Fengchun
    2022, 42 (11):  1562-1568. 
    doi: 10.3969/j.issn.1674-8115.2022.11.007

    Abstract ( 194 )   HTML ( 21 )   PDF (1710KB) ( 35 )  

    Objective ·To explore clinical and pathological characteristics, prognosis and potential related factors of breast cancer patients with brain metastases. Methods ·Seventy-five female breast cancer patients with brain metastases admitted to Renji Hospital, Shanghai Jiao Tong University School of Medicine from January 2007 to December 2021 were selected. The clinical and pathological characteristics of the patients were collected through clinical electronic cases, and the overall survival after brain metastases (OSBM) of the patients was collected during the follow-up period. Receiver operating characteristic curve (ROC curve) was used to evaluate the predictive efficiency of different biochemical factors on OSBM. Kaplan-Meier survival curve was used to analyze the correlation between different clinical and pathological characteristics and patients' OSBM. Cox proportional hazards regression analysis was further performed on statistically significant factors to determine independent influencing factors. Results ·The OSBM of 75 patients was 11.0 (5.0, 24.0) months. The results of ROC curve showed that carbohydrate antigen 125 (CA125) had a good prediction effect on OSBM, and its best cut-off value was 22.3 U/mL. The results of Kaplan-Meier survival analysis showed that the patients whose first metastatic site was brain (P=0.046), whose extracranial disease status was progressed (P=0.042), whose brain metastasis involved one side of tentorium (P=0.013), whose CA125 level<22.3 U/mL (P=0.000), whose treatment methods incladed regional therapy (P=0.034) and whose modified Graded Prognostic Assessment (mGPA) score>1 (P=0.001), had longer OSBM. Cox proportional hazards regression analysis showed that brain metastasis involving both sides of tentorium or meningeal (P=0.029), and CA125 level≥22.3 U/mL (P=0.005) were the risk factors for OSBM, and mGPA score>1 (P=0.033) was the protective factor. Conclusion ·The position of brain metastases in breast cancer, the level of CA125 and the mGPA score are important factors affecting the prognosis of patients.

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    Study on serum vitamin D levels and serum allergens in children with atopic dermatitis
    CHENG Ying, LI Meiyun, CHEN Ji
    2022, 42 (11):  1569-1575. 
    doi: 10.3969/j.issn.1674-8115.2022.11.008

    Abstract ( 204 )   HTML ( 16 )   PDF (1506KB) ( 44 )  

    Objective ·To explore the correlation between serum 25-hydroxyvitamin D [25(OH)D] levels and serum allergens in children with atopic dermatitis (AD), and provide a theoretical basis for the prevention and treatment of AD. Methods ·From November 2017 to February 2019, a total of 165 pediatric patients with AD who visited the Dermatology Department of Shanghai Children′s Medical Center were enrolled in this study. AD severity was assessed according to the severity scoring of atopic dermatitis (SCORAD). Serum total IgE and specific IgE (sIgE) levels of 19 common allergens were detected, and serum25(OH)D was determined by liquid chromatography-tandem mass spectrometry. All data were statistically analyzed. Correlation analysis was conducted by spearman's or point-biserial correlation analysis. Chi-square test was used for comparison of count data between groups. Binary linear regression analysis was used to identify the risk factors. Results ·SCORAD score was shown to be positively correlated with serum total IgE level, absolute value and percentage of eosinophils in children with AD (r=0.213, P=0.009; r=0.268, P=0.001; and r=0.249, P=0.003, respectively). However, there was no significant correlation between serum 25(OH)D concentrations and SCORAD score in children with AD, while there was a significant negative correlation between the serum 25(OH)D concentrations and the total IgE levels, and the percentage of eosinophils in children with AD (r=-0.212, P=0.011; and r=-0.174, P=0.039, respectively) was shown. In this study, the most common allergens with sIgE grade Ⅲ or above were house dust mite, egg white and cashew nut, followed by crab, cat dander, mixed mold, shrimp, dog dander, house dust and mixed tree pollen from high to low. There was no significant correlation between the number of allergen-positive items and serum 25(OH)D concentrations in children with AD (χ2=6.27, P=0.804). However, AD patients with positive house dust mite and mixed mold were statistically significant in different serum [25(OH)D] groups (P=0.000, and P=0.043, respectively). In addition, univariate logistic regression analysis showed that serum [25(OH)D] levels were risk factor for house dust mite allergy in children with AD. The risk of household dust mite allergy in children with serum 25(OH)D deficiency was 8.196 times than that of children with sufficient serum 25(OH)D (OR=8.196, P=0.000). However, serum [25(OH)D] levels were not the risk factor for house dust mite allergy in children with AD. Conclusion ·Serum [25(OH)D] levels might affect the sensitization of children with AD to house dust mite. Avoidance of allergens according to the sIgE test and Vitamin D supplementation might be of great significance in the prevention and treatment of AD.

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    Treatment effect of esomeprazole magnesium combined with hemocoagulase on upper gastrointestinal hemorrhage
    TANG Chuang, ZHANG Xin, ZHANG Ting
    2022, 42 (11):  1576-1581. 
    doi: 10.3969/j.issn.1674-8115.2022.11.009

    Abstract ( 170 )   HTML ( 14 )   PDF (1380KB) ( 26 )  

    Objective ·To observe the total clinical effective rate of esomeprazole magnesium combined with hemocoagulase in the treatment of upper gastrointestinal hemorrhage (UGH) and its influence on upper gastrointestinal mucosa, inflammatory response, stress response and hemorheology. Methods ·A total of 146 patients with UGH, admitted to West China Airport Hospital, Sichuan University from March 2020 to March 2021, were divided into combined group and control group by using the random number table method, with 73 cases in each group. On the basis of conventional treatment, patients in the control group were treated with esomeprazole magnesium, and patients in the combined group were given esomeprazole magnesium combined with hemocoagulase. The total clinical effective rate after 5 d of treatment, as well as serum related factors [serum prostaglandin E2 (PGE2), and 6-keto-prostaglandin F (6-Keto-PGF)], stress indicators [nitric oxide (NO), cortisol (COR), and gastrin (GAS)], serum inflammatory factors [C-reactive protein (CRP), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α)] and hemorheological indicators [whole blood high-shear viscosity, whole blood low-shear viscosity, erythrocyte sedimentation rate, and hematocrit] before and after treatment were compared between the two groups. Results ·The total clinical effective rate of 95.89% in the combined group was higher than 86.30% in the control group (P=0.042). After 5 d of treatment, the stress indicator (NO, COR, and GAS) levels, serum inflammatory factors (CRP, IL-6, and TNF-α), erythrocyte sedimentation rate, and 6-Keto-PGF in the combined group were lower than those in the control group (all P=0.000). The whole blood high-shear viscosity, whole blood low-shear viscosity, hematocrit and PGE2 levels were higher in the combined group compared to the control group (all P=0.000). Conclusion ·The clinical effect of esomeprazole magnesium combined with hemocoagulase is significant in the treatment of UGH, which may be related to the effect of reducing body′s stress response and inflammatory response, changing hemorheology and protecting upper gastrointestinal mucosa.

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    Public health
    Validity and reliability of Chinese version of Occupational Sitting and Physical Activity Questionnaire among intensive care nurses
    HUANG Kanghui, LI Mengyao, SHI Lan, ZHENG Ziyi, WEN Fule, ZHANG Yaqing
    2022, 42 (11):  1582-1588. 
    doi: 10.3969/j.issn.1674-8115.2022.11.010

    Abstract ( 174 )   HTML ( 17 )   PDF (1658KB) ( 33 )  

    Objective ·To introduce and translate the Occupational Sitting and Physical Activity Questionnaire (OSPAQ), and evaluate the reliability and validity of the Chinese version of OSPAQ among intensive care nurses. Methods ·After the translation of OSPAQ, from March 2021 to July 2021, 113 intensive care nurses from two comprehensive tertiary first-class hospitals in Shanghai were conveniently selected as research objects. All participants wore ActiGraph wGT3X-BT accelerometer for 7 consecutive days, and then filled in the Chinese version of OSPAQ. Thirty nurses were retested after an interval of a week. Intraclass correlation coefficient was used to evaluate retest reliability. Criterion validity was assessed by using Spearman′s rank correlation coefficients and Bland-Altman plots. Results ·The sample included 113 intensive care nurses (20 nurses working in day shifts, and 93 nurses working in rotating shifts). The test-retest intraclass correlation coefficients for occupational sitting, standing, and walking in OSPAQ ranged from 0.766 to 0.882, which was evaluated as excellent. The test-retest intraclass correlation coefficients for heavy labour were 0.606, which was evaluated as acceptable. Spearman′s rank correlation coefficients for sitting (0.456) and walking (0.428) showed moderate associations, and that for standing (0.200) showed weak associations. Based on the analysis of the two subgroups, the Spearman′s rank correlation coefficients for sitting and walking showed strong associations among day shifts nurses, while those showed moderate associations among rotating shifts nurses. The Spearman′s rank correlation coefficients for standing showed weak or no associations among day shifts nurses and rotating shifts nurses. Bland-Altman consistency analysis showed that the consistency between the results of sitting posture, standing posture and walking measured by Chinese version OSPAQ and accelerometer was 93.8% to 94.7% in 113 nurses. When using Chinese version OSPAQ self-assessment, the time of sitting posture and standing posture was underestimated and the time of walking was overestimated. Conclusion ·The Chinese version of OSPAQ has good test-retest reliability and moderate validity for occupational sitting and walking among intensive care nurses. It is a simple and reliable assessment tool for occupational physical activity.

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    Techniques and methods
    Detection and analysis of copy number of HER2 gene in breast cancer tissue samples
    SUN Yameng, MA Ye, GUO Runsheng
    2022, 42 (11):  1589-1597. 
    doi: 10.3969/j.issn.1674-8115.2022.11.011

    Abstract ( 174 )   HTML ( 14 )   PDF (2469KB) ( 50 )  

    Objective ·A copy number detection system of HER2 genes in FFPE samples and plasma samples of breast cancer patients was established by using droplet digital PCR system (ddPCR), and then the performance of the detection system was evaluated to provide a scientific basis for clinical auxiliary diagnosis and treatment. Methods ·To evaluate the detection system, 12 tissue samples of breast cancer patients and 4 paired plasma samples were collected from the Department of General Surgery, Jiading District Central Hospital, Shanghai University of Medicine & Health Sciences from January 2020 to June 2021. In the same period, 24 plasma samples of healthy volunteers and 77 tissue sections of breast cancer patients were collected to extract nucleic acids, to establish limit of blank (LoB) and performance evaluation of detection system respectively. The detection system was designed and screened, and subsequently LoB, precision, sensitivity and linear range were also evaluated. We used next generation sequencing (NGS) to verify the consistency of ddPCR detection system, and compared the accuracy of the detection system with fluorescence in-situ hybridization (IHC)/immunohistochemistry (FISH) gold standard method. χ 2 test and matched samples t-test were used for comparison of quantitative data. Poisson probability function was used to analyze the blank detection limit of the system, the coefficient of variation (CV) was used to evaluate the sensitivity and precision, and linear regression correlation coefficient ( R 2 ) was used to evaluate the linear range. Results ·The ddPCR platform was used to establish the quantitative and amplification detection system of HER2 gene copy number in tissue section DNA of breast cancer patients. Compared with the results of tissue nucleic acid and plasma free nucleic acid samples detected by NGS method, the correlation coefficient of copy number detection results was 0.987, and the consistency of amplification was 100%. The intra assay precision and inter assay precision of the evaluation system were 6.8% and 9.4%, respectively. The sensitivity was 1 ng and the linearity was good ( R2>0.98). Compared with the traditional IHC/FISH method, the sensitivity and specificity were 84.6% (95% CI 64.3%~95.0%) and 76.5% (95% CI 62.2%~86.8%), Kappa was 0.57, and the consistency rate was 79.2%. Conclusion ·A copy number detection system for HER2 in breast cancer patients is established by using ddPCR platform, with good consistency in clinical detection, providing auxiliary means for the diagnosis, treatment and prognosis of breast cancer patients.

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    Research progress in application of electrodermal activity at rest in autistic spectrum disorder
    MA Jiayin, WU Yuwei, WU Jiajia, TANG Lin, SHAN Chunlei, XU Jianguang
    2022, 42 (11):  1598-1604. 
    doi: 10.3969/j.issn.1674-8115.2022.11.012

    Abstract ( 164 )   HTML ( 19 )   PDF (1278KB) ( 52 )  

    Autism spectrum disorder (ASD) is a developmental disorder of which the core symptoms are deficits in social communication and interaction, restricted interests as well as repetitive behaviors. The potential cause of core symptoms of ASD may be the presence of abnormal autonomic arousal level in the resting state which can be reflected by electrodermal activity (EDA). Current researches showed that EDA of ASD population was altered from normal healthy population. Additionally, EDA has its clinical potential in differentiating sub-type within ASD population. EDA has its application advantage in ASD population for its objectivity and simple clinical operation. However, the methodology of EDA is not unified and the research results are highly heterogeneous, which indicates the limits of EDA. It is necessary to integrate EDA with other bio-markers and contemporary brain function detection technology, to further reveal the mechanism of abnormal autonomic arousal level and imbalanced autonomic nervous system in the resting state in ASD population. Non-invasive brain stimulation (NIBS) is in main trend of intervention on ASD population, but the standardized and unified clinical intervention has not been formed, which remains to be further studied. Clinical potential of sub-type differentiation by EDA may help to guide accurate NIBS intervention protocol. This paper reviews the application progress of EDA in the resting state in ASD population for the recent 10 years, which is expected to offer theoretical basis for further studying bio-markers and rehabilitation intervention of ASD, and to provide new perspectives for diagnosing and treating ASD.

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    Progress in the correlation between atopic dermatitis and colonization of Staphylococcus
    NURXAT Nadira, LI Min, LIU Qian
    2022, 42 (11):  1605-1611. 
    doi: 10.3969/j.issn.1674-8115.2022.11.013

    Abstract ( 152 )   HTML ( 15 )   PDF (1255KB) ( 55 )  

    Atopic dermatitis (AD) is an inflammatory disease characterized by recurrent itch and skin lesion. The pathological mechanisms of AD include the immunological disorders caused by genetic variants with environment changing and the damage of skin barrier. Recent studies have found that the topical dysbiosis of skin microbiota promotes AD. Compared to the normal skin, the diversity of the microbiome in the lesion skin decreases drastically. Especially, in company with the decreased abundance of protective coagulase-negative Staphylococci (CoNS), the load of Staphylococcus aureus (S. aureus) meets apparent increase. S. aureus is an important clinical pathogen which can cause skin infection, bacteremia, etc. Among AD patients, S. aureus can aggravate the itch and lesion of skin by expressing various toxins, proteases, superantigens, etc. Monitoring the skin flora, especially the characteristics of Staphylococci, provides new ideas for the diagnosis and treatment of AD. Meanwhile, considering the difficulties that the conventional antibiotic therapy failed to reverse and maintain the microbiota of the patient with severe symptom, the application of the commensal microbiome with antimicrobial function provides a new insight into the treatment of AD.

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    Advances in postoperative adjuvant targeted therapy for patients with stage ⅠB-ⅢA non-small cell lung cancer
    LI Ruonan, CHEN Xiaoke, XU Yuanyuan, TAN Qiang
    2022, 42 (11):  1612-1619. 
    doi: 10.3969/j.issn.1674-8115.2022.11.014

    Abstract ( 194 )   HTML ( 16 )   PDF (1339KB) ( 42 )  

    As the cancer with the highest mortality rate in the world, the treatment of lung cancer has always been a difficult problem for a wide range of patients and physicians alike. Based on the degree of differentiation, morphological features and biological characteristics, lung cancer can be divided into small cell lung cancer and non-small cell lung cancer (NSCLC). The incidence of NSCLC accounts for 80%?85%. Clinically, the treatment options of NSCLC include surgery, chemotherapy, radiotherapy, targeted drug therapy, immunotherapy, etc. For the patients with stage ⅠB?ⅢA NSCLC, in addition to the first choice of surgical treatment, postoperative adjuvant therapy is applied to reduce tumor recurrence and metastasis. Studies have shown that targeted drugs are efficient and safe in the adjuvant therapy for NSCLC patients, and the most attention has been given to agents that target mutations in the epidermal growth factor receptor (EGFR) gene, such as epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs). At present, three generations of EGFR TKIs have been approved for clinical use. Among them, the first generation EGFR-TKIs are dominant in the research and application of adjuvant therapy. For example, erlotinib and gefitinib can prolong the disease-free survival (DFS) and overall survival (OS) of patients after surgery, and icotinib has been approved for postoperative adjuvant therapy in China because of its obvious improvement of patients' DFS. Compared with the placebo, the third generation EGFR-TKIs drug osimertinib demonstrated a more significant DFS advantage in the ADAURA trial, decreased tumor recurrence in central nervous system and brought greater benefits in DFS to patients previously treated with standard chemotherapy regime. Osimertinib or chemotherapy combined with osimertinib has therefore become the standard of care for the patients with postoperative adjuvant therapy of stage ⅠB?ⅢA NSCLC. As the third generation EGFR-TKIs new drugs, the clinical trials of almonertinib and furmonertinib for postoperative adjuvant therapy are also underway. This article systematically summarizes the structure of EGFR, the types and detection methods of EGFR gene mutations, introduces the treatment strategies of clinical use of EGFR-TKIs, and discusses the problems that may be encountered in the clinical use of EGFR-TKIs.

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    Diagnosis and treatment of congenital vascular ring: prenatal and postnatal
    GU Shuneng, WU Pengfei, WU Yurong
    2022, 42 (11):  1620-1626. 
    doi: 10.3969/j.issn.1674-8115.2022.11.015

    Abstract ( 176 )   HTML ( 14 )   PDF (1276KB) ( 27 )  

    Congenital vascular ring (CVR) is a rare congenital vascular malformation. During aortic arch development, congenital variations in vessels may occur, some of which form vascular rings that cause compression around the airway and/or esophagus, causing a range of respiratory and gastrointestinal symptoms. In addition, CVR may be associated with a variety of chromosomal and genetic abnormalities and other intracardiac and extracardiac structural malformations. The treatment and prognosis varies from type to type according to differences in anatomic structure and degree of compression. With the standardization and popularization of prenatal screening of fetal congenital diseases, the liversification of postnatal assessment methods and application of new operation methods, the prenatal detection rate of CVR has been increased, and the postnatal evaluation has been more accurate. The postoperative prognosis of some diseases has been improved. However, the clinical cognition of CVR still remains to be improved. The lack of professional consultation on treatment and prognosis may cause excessive anxiety of gravida and families, and even unnecessary induction. The surgical indication, treatment and prognosis of each type of CVR still remain to reach a consensus. This review summarizes the anatomical characteristics, diagnostic methods, clinical manifestations, and treatment and prognosis of CVR types in order to improve and update the understanding of this disease, guide gravidae to make more reasonable decisions during pregnancy and provide reference for diagnosis, treatment and follow-up plan.

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    Research progress of cytokines in treatment of osteoarthritis
    DONG Yushan, ZHANG Wenjie
    2022, 42 (11):  1627-1632. 
    doi: 10.3969/j.issn.1674-8115.2022.11.016

    Abstract ( 234 )   HTML ( 18 )   PDF (1290KB) ( 67 )  

    Osteoarthritis (OA) is a common joint disease in clinic. OA mostly involves the load-bearing joints of the lower limbs, which is easy to cause joint deformity and affect the quality of life after the disease. At present, early treatment of OA focuses on symptomatic treatment and slowing down the wear of articular cartilage. Various cytokines play an important role in the development of OA. Among the interleukin (IL) family, IL-1β/6/8 are associated with the occurrence and progression of OA and are major inflammatory factors. IL-4/10 play a protective role in the joints as an anti-inflammatory component. Tumor necrosis factor-α is also a major component that promotes joint inflammation. In vitro and in vivo studies show that transforming growth factor-β,insulin-like growth factor, and fibroblast growth factor (FGF) can delay the progression of OA. By investigating the roles of the relevant cytokines, we can not only gain insight into the pathogenesis of OA, but also provide new ideas for the treatment of OA. Cytokines are expected to solve the dilemma of suboptimal efficacy of current traditional treatments for OA. At present, many beneficial research results are achieved in the field of cytokine application to the prevention and treatment of OA, and a part of them begins to enter clinical trials and applications. This review summarizes the research progress and partial mechanisms of several classes of cytokines with potential effects on OA.

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    Summary of clinical research of Pediococcus pentosaceus in treatment of infantile colic
    WU Shiyin, CAI Meiqin
    2022, 42 (11):  1633-1637. 
    doi: 10.3969/j.issn.1674-8115.2022.11.017

    Abstract ( 476 )   HTML ( 16 )   PDF (1193KB) ( 104 )  

    Infantile colic is a common disease. Its etiology is not clear, which may be related to the imbalance of intestinal flora. Pediococcus pentosaceus is a probiotic of Pediococcus of Lactobacillaceae. It can enhance host immunity and improve the diversity of intestinal flora. This paper will introduce the characteristics of probiotics such as antibacterial capability, intestinal colonization ability and antibiotic sensitivity of Pediococcus pentosaceus, and the probiotic functions such as antioxidant, antiviral and immune regulation, and elaborate on the possible pathogenesis of infantile colic, the efficacy and potential mechanism of Pediococcus pentosaceus in the treatment of infantile colic, and the current situation of clinical research at home and abroad, in order to provide a scientific basis for Pediococcus pentosaceus in the treatment of infantile colic.

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    Brief original article
    Efficacy of modified coronally advanced tunnel technique combined with resin restoration for the treatment of multiple gingival recession with non-carious cervical lesions
    LIAO Yue, DONG Jiachen, SONG Zhongchen
    2022, 42 (11):  1638-1643. 
    doi: 10.3969/j.issn.1674-8115.2022.11.018

    Abstract ( 153 )   HTML ( 13 )   PDF (2981KB) ( 27 )  

    Objective ·To observe the clinical efficacy of modified coronally advanced tunnel technique (MCAT) combined with resin restoration in treating multiple gingival recession with non-carious cervical lesion (NCCL). Methods ·Subjects with multiple gingival recession and NCCL were enrolled in the trial. All the patients were treated with MCAT with subepithelial connective tissue graft (sCTG) and resin restoration to repair the soft and hard tissue defect. Probing depth (PD), bleeding index (BI), plaque index, (PLI), recession height (RH), keratinized tissue width (KTW), gingival thickness (GT), and interdental papilla height were measured at baseline and 6-month follow-up. Root coverage esthetic score (RES) was also evaluated 6 months postoperatively. Results ·Three patients were treated in this study, including 9 sites. The mean BI, PLI and RH were significantly decreased at 6 months after treatment compared with the baseline. The augment of KTW, GT and interdental papilla height was achieved at follow-up, and interdental papilla height increased by (0.67±0.75) mm compared with the baseline. The mean RES was 8.56±1.33, postoperatively. Conclusion ·The MCAT combined with resin restoration could be a promising treatment for multiple gingival recession, resulting in ideal esthetic effect by the augment of buccal and interdental soft tissue.

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    Case report
    A case of neonatal MIRAGE syndrome caused by novel mutation of SAMD9 gene
    LI Yahui, WANG Yiwen, XIE Lijuan
    2022, 42 (11):  1644-1648. 
    doi: 10.3969/j.issn.1674-8115.2022.11.019

    Abstract ( 232 )   HTML ( 12 )   PDF (2083KB) ( 45 )  

    A female newborn, delivered by cesarean section for oligohydramnios and severe intrauterine growth retardation at gestational age of 30 weeks. She presented progressive severe respiratory distress and symptoms of adrenal insufficiency, such as hyperpigmentation, hypotension, disorder of electrolyte and glucose metabolism soon after birth. The newborn died at the fifth day because of multiple organ failure. Gene analysis showed that there was heterozygous variant in the SAMD9 gene of the newborn (c.4598G>A, p.Arg1533Gln) and she was diagnosed with MIRAGE syndrome (myelodysplasia, infection, restriction of growth, adrenal hypoplasia, genital phenotypes, and enteropathy). MIRAGE syndrome is usually complex and critical with multisystem disorder. Early identification with genetic testing can help diagnosis and guide treatment, which can improve the prognosis.

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