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    Basic research
    Effect of enzyme digestion into cell clumps on protein levels of OCT4 and SOX2 in human embryonic stem cells
    Xiao-zhi SUN, Shuang LI, Ying JIN, Bing LIAO
    2021, 41 (4):  413-420. 
    doi: 10.3969/j.issn.1674-8115.2021.04.001

    Abstract ( 227 )   HTML ( 20 )   PDF (3246KB) ( 110 )  
    Objective

    · To reveal the effect of enzyme digestion into cell clumps on protein levels of octamer-binding protein 4 (OCT4) and sex determining region Y-box 2 (SOX2) in the human embryonic stem cells (hESCs) and its possible molecular mechanism.

    Methods

    · ① Western blotting was conducted to detect the protein levels of OCT4 and SOX2 in the hESCs treated by enzyme digestion into cell clumps [Dispase or collagenase type Ⅳ (COL4)], enzyme digestion into single cells (Accutase or trypsin), or mechanical scraping dissociation. ② Ethylene glycol tetraacetic acid (EGTA) was used to destruct hESC-hESC interaction, and protein levels of OCT4 and SOX2 in the hESCs treated with EGTA alone or combined with COL4 were detected by Western blotting. ③ Quantitative proteomics analysis based on mass spectrometry was carried out to identify differentially expressed total proteins and phosphorylated proteins in the hESCs treated with COL4 alone or in combination with EGTA. ④ hESCs were pre-treated with proteasome inhibitor (bortezomib) and lysosome inhibitor (chloroquine), respectively, followed by Western blotting and immunofluorescence staining to measure their effects on COL4-induced down regulation of OCT4 and SOX2. ⑤ Western blotting analysis was conducted to test the effect of COL4 treatment on cofilin as well as Rac1 inhibitors (EHT1864 and EHop-016) treatment on the protein level of SOX2 in the hESCs.

    Results

    · ① COL4 or Dispase treatment led to protein levels decrease of OCT4 and SOX2, while Accutase, trypsin and mechanical dissociation did not elicit such effect. ② EGTA rescued OCT4 and SOX2 protein levels decrease indcued by COL4 treatment. ③Quantitative mass spectrometry and functional enrichment analysis suggested that microtubule, actin filament (F-actin) and lysosome-associated proteins might be related to OCT4 and SOX2 protein levels decline in the suspended clumps of hESCs upon COL4 treatment. ④Chloroquine, instead of bortezomib, could block COL4-induced protein levels decrease of OCT4 and SOX2. ⑤The treatment of COL4 or Rac1 inhibitors could downregulate protein levels of cofilin and SOX2.

    Conclusion

    · Upon hESC detachment in cell clumps induced by enzyme treatment, the protein levels of SOX2 and OCT4 decline, which may be related to Rac1/cofilin/F-actin signaling pathway and lysosome.

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    Single-cell RNA sequencing reveals the spatio-temporal expression profile of SARS-CoV-2 related receptor in human and mouse testes
    Jia-qiang LUO, Liang-yu ZHAO, Chen-cheng YAO, Zi-jue ZHU, Xiao-yu XING, Peng LI, Ru-hui TIAN, Hui-xing CHEN, Jie SUN, Zheng LI
    2021, 41 (4):  421-426. 
    doi: 10.3969/j.issn.1674-8115.2021.04.002

    Abstract ( 297 )   HTML ( 17 )   PDF (4839KB) ( 206 )  
    Objective

    ·To analyze the spatio-temporal expression profile of angiotensin-converting enzyme 2 (ACE2), the receptor of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), in human and mouse testes based on single-cell RNA sequencing (scRNA-Seq).

    Methods

    ·Ten testicular tissues from humans and nine testicular tissues from C57BL/6 mice with normal developmental stages were collected and digested into single cell suspensions by enzyme, and then the cell-gene expression matrixes were obtained by scRNA-Seq standard processing procedures. After quality control, data standardization, batch effect processing, clustering, and dimensionality reduction, each subgroup of cells was annotated based on known testicular cell bio-markers to clarify the expression patterns and differences of ACE2 in human and mouse testes with normal developmental stages.

    Results

    ·In this study, nine testicular cell subgroups found in human and mice were identified, including three subgroups of germ cells (spermatogonia, spermatocytes, and spermatids/sperm) and six subgroups of somatic cells (Sertoli cells, macrophages, vascular smooth muscle cells, endothelial cells, Leydig cells, and peritubular myoid cells). In terms of spatial distribution, ACE2 was predominantly expressed in human Sertoli cells in adult testes, and also expressed in Leydig cells, peritubular myoid cells, and germ cells. In terms of time scale, the transcription abundance of ACE2 in human Sertoli cells increased with the development of testis, and the expression level of ACE2 in Sertoli cells after puberty was significantly higher than that in infancy and childhood (P=0.000). Judging from the stages of mouse testicular development, the expression patterns of Ace2 were both significantly different from those in humans. In testis of the 5-week-old adult C57BL/6 mouse, the transcription level of Ace2 was low and it was mainly expressed in vascular smooth muscle cells (P=0.000), while the number of Ace2 positive cell in Sertoli cells was extremely low.

    Conclusion

    ·SARS-CoV-2 may mainly infects human testis through Sertoli cells, and the conventional C57BL/6 mouse model is not suitable to simulate the effect of SARS-CoV-2 infection on human testicular function.

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    Molecular mechanism of apoptosis-inducing factor regulated by FBXO22 and its role in cancer cell apoptosis
    Meng-di LIU, Yi-lian PAN, Xiao-na ZHU, Shuo YANG, Qian YANG, Yun YU
    2021, 41 (4):  427-433. 
    doi: 10.3969/j.issn.1674-8115.2021.04.003

    Abstract ( 263 )   HTML ( 20 )   PDF (2613KB) ( 144 )  
    Objective

    ·To explore the substrate of F-box only protein 22 (FBXO22) and its mechanism, and further study the role of FBXO22 in cancer cell apoptosis.

    Methods

    ·The interaction protein of FBXO22, namely apoptosis-inducing factor (AIF), was searched and identified by immunoprecipitation (IP)-coupled liquid chromatograph mass spectrometer/mass spectrometer (LC-MS/MS) technology. The effect of FBXO22 on ubiquitination of AIF was detected by ubiquitin modification assay under denatured conditions. Short hairpin RNA (shRNA) knockdown FBXO22 and overexpression FBXO22 were used to detect the expression of AIF. The effect of FBXO22 knockdown or overexpression on apoptosis of colon cancer cells was detected by flow cytometry.

    Results

    ·IP-coupled LC-MS/MS technology showed that FBXO22 could interact with AIF, and ubiquitin modification assay under denaturing conditions showed that FBXO22 could mediate ubiquitination of AIF. Knockdown of FBXO22 could up-regulate the level of AIF, while overexpression of FBXO22 could down-regulate the level of AIF. The results of flow cytometry showed that knockdown of FBXO22 could promote the apoptosis of colon cancer cells induced by N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), while overexpression of FBXO22 could inhibit the apoptosis of colon cancer cells induced by MNNG.

    Conclusion

    · FBXO22 regulates AIF-mediated apoptosis of colon cancer cells by ubiquitination and degradation of AIF.

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    USP47 induces cisplatin resistance in head and neck squamous cell carcinoma by up-regulation of anti-apoptotic proteins
    Tong TONG, Xing QIN, Fei XIE, Ying-ying JIANG, Jian-bo SHI, Jian-jun ZHANG
    2021, 41 (4):  434-441. 
    doi: 10.3969/j.issn.1674-8115.2021.04.004

    Abstract ( 258 )   HTML ( 17 )   PDF (3685KB) ( 142 )  
    Objective

    · To investigate the effects and underlying mechanism of ubiquitin carboxyl-terminal hydrolase 47 (USP47) on cisplatin resistance in head and neck squamous cells carcinoma (HNSCC).

    Methods

    · The cisplatin-resistant cell lines of HNSCC were constructed by gradient increase of cisplatin concentration. The expression of USP47 in cisplatin-resistant cell was analyzed by real-time PCR and Western blotting. Stable USP47 over-expressed and silenced USP47 HNSCCs cell lines were generated by lentivirus vector. The influence of the increased or silenced USP47 expression on cisplation resistance were tested by MTT. The influence of USP47 on cell proliferation, colony formation, and apoptosis were examined. Western blotting was performed to detect the expression and ubiquitination levels of X-linked inhibitor of apoptosis protein (XIAP) and recombinant human B-cell lymphoma factor 2 xL (Bcl-xL).

    Results

    · USP47 expression was significantly increased in cisplatin-resistant cancer cells. Over-expression of USP47 significantly increased the half maximal inhibitory concentration (IC50), while knockout of USP47 gene increased the sensitivity to cisplatin. In addition, the data showed that USP47 expression was negatively related to the proliferation and colony formation abilities of HNSCC cell lines, but it remarkably enhanced the anti-apoptotic effect of HNSCC cells. The protein levels of XIAP and Bcl-xL were significantly elevated in stable USP47 over-expressed cell lines and ubiquitination assay proved that USP47 reduced the ubiquitination levels of XIAP and Bcl-xL proteins. Knockdown of the XIAP and Bcl-xL gene expression blocked cisplatin resistance induced by USP47 gene overexpression.

    Conclusion

    · USP47 can increase the protein expressions of XIAP and Bcl-xL by inhibiting the ubiquitination modification of them, thereby mediate the cisplatin resistance of HNSCC cells. Inhibition of USP47 expression provides potential application value to reduce cisplatin-resistance during the treatment on HNSCC.

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    Correlation analysis of salivary microbiome and host blood lipid levels
    Fen ZHAO, Qiao-qiao JIN, Ke-yong YUAN, Xiu-xiu HOU, Zheng-wei HUANG, Rui MA
    2021, 41 (4):  442-447. 
    doi: 10.3969/j.issn.1674-8115.2021.04.005

    Abstract ( 223 )   HTML ( 16 )   PDF (4081KB) ( 99 )  
    Objective

    ·To analyze the correlation between salivary microbiome and host blood lipid levels.

    Methods

    ·Samples of saliva from 114 volunteers aged 45?60 years were collected to extract total DNA, and then 16S rDNA V3?V4 region was amplified with polymerase chain reaction. After the amplified products were sequenced via Illumina MiSeq PE300 platform,the obtained sequences were subjected to operational taxonomic units (OTUs) clustering and species annotation.The Spearman correlation analysis was also carried out to find the correlations between the top 50 abundant genera in the salivary microbiome and host blood lipid levels, P<0.05 was considered as statistically significant difference.

    Results

    ·The saliva samples of 114 volunteers were sequenced with an average sequence number of 41 084±4 740, and the number of OTUs obtained by clustering was 1 153, annotated to 23 phyla, 43 classes, 89 orders, 147 families and 317 genera. Species clustering heat map of 114 samples suggested that salivary microbiome has similar abundance patterns of species composition in the population.Spearman correlation analysis showed that among triacylglycerol (TAG) the top 50 abundant genera in the salivary microbiome, Neisseria spp. were negatively correlated with the host serum total cholesterol (TCH), and low density lipoprotein cholesterol level (LDL-Ch); Gemella spp. are positively correlated with Host serum TCH and LDL-Ch; high-density lipoprotein cholesterol presents negative correlation with Comamonas spp., Filifactor spp. and Parvimonas spp.

    Conclusions

    ·The composition structure of salivary microbiota in the population is quite stable and is related to the levels of host blood lipids. It is promising to further explore salivary microbiota as early warning indicators of individual lipid metabolism abnormalities.

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    Association of alternative splicing and tumor immune in gastric cancer based on TCGA data set
    Qi-sheng GU, Mi-li ZHANG, Can CAO, Ji-kun LI
    2021, 41 (4):  448-458. 
    doi: 10.3969/j.issn.1674-8115.2021.04.006

    Abstract ( 260 )   HTML ( 8 )   PDF (4501KB) ( 117 )  
    Objective

    · To investigate the association of alternative splicing and tumor immune in gastric cancer based on The Cancer Genome Atlas (TCGA) data set.

    Methods

    · Transcriptomic data, genomic data and corresponding clinical information of 375 tumor tissues and 32 paired adjacent normal tissues from gastric cancer patients were separately downloaded from TCGA portal. Percent-spliced-in matrix containing 452 gastric cancer tissues and paired normal tissues were downloaded from Spliceseq database. The microarray transcriptomic data GSE15459 including 192 gastric cancer patients were downloaded from Gene Expression Omnibus (GEO). ConcensusClusterPlus package was used for classifying the samples based on splicing events. Competitive Gene Set Test Accounting for Inter-gene Correlation (CAMERA) and Gene Set Variation Analysis (GSVA) were used to apply pathway and gene set analysis. CIBERSORT was used to interrogate the condition of tumor microenvironment of samples through deconvolution of profiling of mRNA expression.

    Results

    · Four hundred and forty-five gastric cancer tissues and paired adjacent normal tissues were included into the study after filtering, involving 4 051 genes and 8 649 splicing events. There were aberrant splicing events existing in gastric cancer tissues compared to adjacent normal tissues. The gastric cancer samples could be divided into 4 subtypes based on different high-frequency splicing events. Many clinical characteristics like T stage, M stage, age, Lauren classification, pathological stage and histological stage among the 4 subtypes were significantly different (P<0.05). The tumor hallmark characteristics and condition of microenvironment were also marked different among the 4 subtypes. The high expression of certain splicing factors were responsible for clustering of the subtypes (log2FC > 1 and FDR < 0.05), and gene set of core splicing factors was strongly correlated to antigen presentation in tumor (Pearson R = 0.44, P = 0.000).

    Conclusion

    · The variation of splicing events was closely related to clinical characteristics and tumor microenvironment in gastric cancer. Expression of splicing factors dominates the variation of alternative splicing events. Certain splicing factors were expected to be biomarkers for classification of gastric cancer, and targets for improving the efficacy of immunotherapy.

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    Development and performance evaluation of indirect ELISA kit with recombinant leptospira protein as coating antigen
    Jie QIAN, Jian YANG, Yue ZHANG, Rong ZHANG, Xiang-yan ZHANG, Xiao-kui GUO
    2021, 41 (4):  459-466. 
    doi: 10.3969/j.issn.1674-8115.2021.04.007

    Abstract ( 210 )   HTML ( 3 )   PDF (1762KB) ( 114 )  
    Objective

    ·To develop an indirect enzyme-linked immunosorbent assay (ELISA) kit for the detection of leptospira-specific antibodies and evaluate its performance.

    Methods

    ·The highly conserved outer membrane protein in the epidemic strains of the main pathogenic leptospira in China was selected and expressed by prokaryotic expression system. The recombinant protein was purified and used as coating antigen for mass spectrometry identification, and its antigenicity was detected. Serum samples from 40 patients with leptospirosis, 50 patients with other infectious diseases and 391 healthy controls were selected as test samples. The positive control, positive reference and high-titer mixed positive serum for internal control of the kit were prepared from the serum of patients with leptospirosis, the negative control and reference for internal control of the kit were prepared from the serum of healthy controls, and the weak positive control (critical reference) and sensitivity reference for internal control of the kit were prepared from high-titer mixed positive serum. One strong positive serum and one weak positive serum selected from the serum of patients with leptospirosis were used as strong positive and weak positive precision reference for internal control of the kit. The detection system of the indirect ELISA kit was established, its preparation process and reaction conditions were optimized, and its precision, sensitivity, specificity and stability were evaluated.

    Results

    ·The purified recombinant outer membrane protein of leptospira was identified as the target protein by mass spectrometry. The results of performance evaluation showed that the intra assay precision and inter assay precision of the indirect ELISA kit were less than 15% and 20%, respectively, and the sensitivity and specificity were 95.0% and 95.0%, respectively. The performance of indirect ELISA kit was still stable after 12 months' storage at 2?8 ℃.

    Conclusion

    ·The specificity, sensitivity and other performance indicators of the leptospira antibody detection kit meet the requirements of the in vitro test kit, which may be suitable for clinical auxiliary diagnosis, sentinel monitoring of leptospirosis and epidemiological investigation.

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    Inhibition of semen-derived enhancer of viral infection by superoxide dismutase
    Meng-jie QIU, Zhao-feng LI, Yi-bin LI, Yu-liu CHEN, Hong-yan CHENG, Shu-wen LIU, Sui-yi TAN
    2021, 41 (4):  467-472. 
    doi: 10.3969/j.issn.1674-8115.2021.04.008

    Abstract ( 226 )   HTML ( 1 )   PDF (2608KB) ( 138 )  
    Objective

    ·To explore the inhibitory effect of superoxide dismutase (SOD) on the formation of semen-derived enhancer of viral infection (SEVI) amyloid fibrils.

    Methods

    ·Prostatic acid phosphatase 248-286 (PAP248-286) at 440 μmol/L or seminal fluid was incubated with SOD at different activity levels, and at different time points, aliquots were taken from each sample for thioflavin T (ThT) staining, circular dichroism (CD) and transmission electron microscope (TEM) to detect the effect of SOD on the formation of SEVI. The effect of SOD on the ability of SEVI to promote human immunodeficiency virus-1 (HIV-1) infection was detected by virus infection enhancement experiment. The effects of inactivated SOD on the formation of SEVI and the ability of SEVI to promote virus infection were also detected. The SOD activity levels in the semen of five healthy volunteers were detected by using total SOD activity detection kit.

    Results

    ·The results of ThT staining showed that 46 U/mL and 92 U/mL SOD could inhibit the formation of SEVI fibrils in the PAP248-286 solution in vitro at 48-h time point. The results of CD showed that SOD could inhibit the formation of β-sheet structure of SEVI. The results of TEM also showed that SOD could inhibit the formation of SEVI. The ThT staining in the semen also demonstrated the inhibitory effect of SOD on the formation of SEVI. As shown in the virus infection enhancement experiment, SOD could inhibit the promoting effect of SEVI on HIV-1 infection. When SOD was inactivated, the effects above almost disappeared. The SOD activities in the semen of the five healthy males were 74.87, 68.69, 85.46, 113.29, and 109.53 U/mL, respectively.

    Conclusion

    ·SOD can inhibit the formation of SEVI fibrils in vitro and the promoting effect of SEVI on HIV-1 infection.

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    Clinical research
    Effect of metformin on infection event reduction in patients with systemic lupus erythematosus:a post-hoc analysis of a Met-Lupus Trial
    Shi-kai GENG, Le ZHANG, Hui-jing WANG, Liang-jing LÜ, Wei-guo WAN, Fang-fang SUN, Shuang YE
    2021, 41 (4):  473-478. 
    doi: 10.3969/j.issn.1674-8115.2021.04.009

    Abstract ( 230 )   HTML ( 2 )   PDF (1341KB) ( 96 )  
    Objective

    ·To evaluate the effect of metformin on reducing infection events in the systemic lupus erythematosus (SLE) patients with moderate/low disease activity based on a multicenter, randomized, double-blind, placebo-controlled clinical study (Met-Lupus Trial).

    Methods

    ·The 140 participants in the Met-Lupus Trial were randomly divided into the metformin group (67 cases) and the placebo group (73 cases). The metformin tablets or placebo tablets were added to their standard therapy with target dose of 1 500 mg/d, three times per day. The infection events during the 12 months' follow-up of the patients were recorded, including the types of infection events, infection duration, infection severity, and laboratory results during infection. The clinical characteristics between the patients with or without infection as well as between the infected patients treated with metformin or placebo were compared. Multivariate Logistic regression analysis was used to analyze the correlation between metformin and infection events, and survival analysis was used to compare the infection-free survival time between the metformin group and the placebo group.

    Results

    ·By 12 months of follow-up, the exposure rate of metformin in the patients without infection (65.9%) was significantly higher than that in the patients with infection (34.7%, P=0.022), while other clinical parameters were comparable. Multivariate Logistic regression analysis suggested that the use of metformin was an independent protective factor against infection in the SLE patients (OR=0.423, P=0.033). In the infected patients, the severe infection incidence in the metformin group was numerically lower than that in the placebo group, but there was no significant difference (5.9% vs 12.5%, P=0.466). Further analysis showed that the infection duration [7.0 (6.0, 11.8) d] of the metformin group was significantly lower than that of the placebo group [10.0 (7.0, 21.8) d] (P=0.034); meanwhile, the C-reactive protein in the metformin group [2.5 (2.4, 6.4) mg/L] was also lower than that in the placebo group [4.5 (2.5, 8.9) mg/L] without significant difference (P=0.075). Survival analysis showed that infection-free survival of the metformin group was significantly longer than that of the placebo group (HR=0.527, 95%CI 0.294?0.945, P=0.036).

    Conclusion

    ·Metformin may have a potential effect on infection event reduction in the SLE patients with moderate/low disease activity.

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    Procedural manipulation under anesthesia for joint adhesion after trauma of knee operation: report of 21 cases
    Xin JIANG, Ying HE, Bin CAI, Shuai FAN, Xiao-yan LI, Zhi-bin KANG
    2021, 41 (4):  479-482. 
    doi: 10.3969/j.issn.1674-8115.2021.04.010

    Abstract ( 269 )   HTML ( 1 )   PDF (1047KB) ( 132 )  
    Objective

    · To investigate the clinical efficacy and safety of procedural manipulation under anesthesia (MUA) for joint adhesion after trauma of knee operation.

    Methods

    · Twenty one patients with joint adhesion after trauma of knee operation (including 6 cases after reconstruction and/or repair of multiple ligament injury of knee joint, 9 cases after internal fixation of fracture around the knee joint, and 6 cases after operation of patellar dislocation) were treated with procedural MUA. The knee flexion angle was evaluated before, during and after procedural MUA (2, 12 and 24 weeks) with joint protractor. The pain degree was evaluated by visual analogue scale (VAS) within 2 weeks after procedural MUA. The possible complications were recorded during and after procedural MUA.

    Results

    ·Before, during and after procedural MUA (2, 12 and 24 weeks), the average knee flexion angle was 73.8°, 119.3°, 101.7°, 122.5° and 127.4°. Compared with preoperative, the average knee flexion angle increased 53.6° after procedural MUA (24 weeks). VAS evaluation results showed that the VAS score of patients in active state was more than 4 points only 1?3 days after operation. No complications occurred during and after the operation.

    Conclusion

    · Procedural MUA is relatively safe and effective in the treatment of joint adhesion after trauma of knee operation.

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    Association between blood lipid profile in early pregnancy and risk of preeclampsia: a study based on real world data
    Fang ZHANG, Xiao-jin WANG, Jue MA, Yun-ting HE, Hao HE, Jing ZHAI, Yu-na GUO, Yan CHEN, Bing-shun WANG
    2021, 41 (4):  483-488. 
    doi: 10.3969/j.issn.1674-8115.2021.04.011

    Abstract ( 202 )   HTML ( 4 )   PDF (1152KB) ( 101 )  
    Objective

    ·To investigate the association between the blood lipid levels in early pregnancy and the risk of preeclampsia (PE).

    Methods

    ·Pregnant women who underwent routine prenatal examination and delivered in the International Peace Maternity & Child Health Hospital, Shanghai Jiao Tong University School of Medicine from January 2015 to December 2017 were recruited as the study participants. After screening, 26 230 pregnant women were selected, including 680 pregnant women with PE (PE group) and 25 550 pregnant women without PE (control group). The general clinical data and the blood lipid index during 10?14 weeks of pregnancy were obtained from the electronic medical record system. Multivariate Logistic regression analysis was used to explore the relationship between blood lipid levels at different categorical (by quartile) in early pregnancy and the risk of PE.

    Results

    ·Compared with the control group, the levels of total cholesterol (TC), triacylglycerol (TAG), low density lipoprotein (LDL) and lipoprotein B (ApoB) in the early pregnancy of the PE group were higher (all P<0.05), while the level of high density lipoprotein (HDL) was lower (P=0.000). Multivariate Logistic regression analysis showed that TAG (>1.45 mmol/L), LDL (>3.26 mmol/L) and ApoB (>0.86 g/L) were the independent risk factors for PE (all P<0.05), and HDL (>1.99 mmol/L) was the independent protective factor for PE (P=0.002). After adjusting for potential confounding factors, the risk of PE increased successively with the increase of the TAG, LDL and ApoB levels, HDL did the opposite (all P for trend<0.05). With the lowest quartile as a reference, the adjusted OR values and 95% CIs for the highest quartiles of TAG, LDL, ApoB and HDL were 1.90 (1.47?2.45), 1.39 (1.08?1.78), 1.51 (1.13?2.04) and 0.66 (0.50?0.86), respectively.

    Conclusion

    ·Dyslipidemia in early pregnancy indicates an increased risk of PE during pregnancy. Examination of blood lipid level during early pregnancy is helpful for early prediction of PE.

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    Different expression of prolactin in unexplained recurrent pregnancy loss and normal early pregnancy
    Chao CHEN, Cong-cong LI, Feng GUO, Qiao-hong WANG, Ai-min ZHAO
    2021, 41 (4):  489-496. 
    doi: 10.3969/j.issn.1674-8115.2021.04.012

    Abstract ( 192 )   HTML ( 1 )   PDF (3475KB) ( 90 )  
    Objective

    ·To investigate the difference of prolactin (PRL) expression in peripheral blood and decidual stromal cells (DSCs) between unexplained recurrent pregnancy loss (URPL) and normal early pregnancy.

    Methods

    · Eighty cases of URPL patients in our hospital from March 2018 to March 2019 were recruited. Seventy women with induced abortion and 190 normal pregnancy women were selected as controls. Fasting peripheral blood and decidua tissues were collected. The level of PRL in peripheral blood was detected by enzyme-linked immunosorbent assay. Quantitative real time PCR (RT-qPCR), Western bloting (WB) and immunohistochemistry (IHC) were used to detect the PRL expression of DSCs. Independent sample t test was used to compare the data of the two different groups, and one-way analysis of variance test was used to compare the data of the groups.

    Results

    · In the URPL group, 21 cases were excluded for chromosomal abnormality, and 59 cases were finally included. There were 70 cases in the induced abortion group. One hundred and seventy-seven cases in the normal pregnancy group completed follow-up, of which 157 cases were classified as final live birth in the normal pregnancy group, and the rest 20 cases subsequently had pregnancy loss, of which 19 miscarriages occurred in the first 10 weeks of gestation, classified as pregnancy loss in the normal pregnancy group. The PRL levels of peripheral blood in the final live birth in the normal pregnancy group and induced abortion group were similar, with no significant difference between the two groups in the same gestation, and they all increased significantly with advancing gestation (normal pregnancy with final live birth group: 5?5+6 week vs 6?7+6 week, P = 0.002; 6?7+6 week vs 8?9+6 week,P = 0.012;Induced abortion group: 5?5+6 week vs 6?7+6 week, P = 0.015; 6?7+6 week vs 8?9+6 week, P = 0.023). The levels of PRL in the peripheral blood of patients with URPL had no significant change with advancing gestation, and they were significantly lower than those of women with final live birth in the normal pregnancy group and induced abortion women in the same gestation from the 6th week (6?7+6week: P = 0.012, 0.024; 8?9+6 week: P = 0.015, 0.003). In addition, the levels of PRL in the peripheral blood of the women in the normal pregnancy group who later suffered miscarriage were similar to those in the URPL group of the same gestation. The PRL expression of DSCs in the induced abortion group increased significantly in both RNA and protein level with advancing gestation (5?5+6 week vs 6?7+6 week: RT-qPCR, P = 0.020; WB, P = 0.010; IHC,P = 0.030. 6?7+6 week vs 8?9+6 week: RT-qPCR, P = 0.011; WB, P = 0.034; IHC, P = 0.012). However, the increase of PRL expression of DSCs in the URPL group was delayed: the PRL expression of DSCs in women with 6?7+6week and 8?9+6 week of gestation was significantly lower than that in women accepting induced abortion of the same gestation (6?7+6 week: RT-qPCR, P = 0.012; WB, P = 0.014; IHC,P = 0.028. 8?9+6 week: RT-qPCR, P = 0.011; WB, P = 0.030;IHC, P = 0.026).

    Conclusion

    · PRL expression of the decidua is defective. The PRL expression of DSCs in URPL patients is deficient, and the lack of decidualization of DSCs may be related to the occurrence of URPL. The low level of PRL in peripheral blood after 6 weeks of gestation may indicate embryonic dysplasia.

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    Correlation between sleep quality and everyday cognitive function in perimenopausal and postmenopausal women
    Wen-jun HUANG, Su-su JIANG, Min-fang TAO
    2021, 41 (4):  497-501. 
    doi: 10.3969/j.issn.1674-8115.2021.04.013

    Abstract ( 216 )   HTML ( 2 )   PDF (1111KB) ( 104 )  
    Objective

    · To evaluate the correlation between sleep quality and everyday cognitive function in perimenopausal and postmenopausal women.

    Methods

    · Two hundred and eighty one women who visited the menopause clinic of Shanghai Sixth People's Hospital for the first time were included in the study. Socialdemographic characteristics of the patients were obtained by using the general information questionnaire. Pittsburgh Sleep Quality Index (PSQI) was used toassess sleep quality and its seven dimensions (sleep latency, sleep duration, sleep efficiency, sleep disturbance, subjective sleep quality, sleep medicationand daytime dysfunction).Everyday cognitive function was assessed by using the short version of Everyday Cognition (Ecog-12). Mann-Whitney U (two independent samples) or Kruskal-Wallis H (multiple indepentdent samples) test was used for univariate analysis of patents' everyday cogitive function.Spearman rank correlation was used to analyze the relationship of sleep quality and its seven dimensions with everyday cognitive function.Multiple linear stepwise regressionwas used to analyze the influencing factors of everyday cognitive function.

    Results

    · The Ecog-12 score of 281 women was 1.42 (1.17,1.67). The score of PSQI was 6.0 (3.0, 10.0). 40.9% of perimenopausal and postmenopausal women had poor sleep quality (PSQI≥8). There was no statisticaldifference in patients' everyday cognitive function among all social demographic characteristic variables (age, body mass index, monthly income, occupationstatus, educational level, menopausal status, parity, regular exercise and history of chronic diseases). Compared to women with good sleep quality,women with poor quality scored higher in Ecog-12 (P=0.000). Spearman correlation analysis showed that the scores of 7 dimensions of PSQI and totalscore of PSQI were positively correlated with Ecog-12 score (P=0.000). Multiple linear regression analysis showed that the explanation rate of sleep medication and daytime dysfunction on everyday cognitive function deline was 12.6%.

    Conclusion

    · Sleep medication and daytime dysfunction are the independent influencing factors of everyday cognitive function decline in perimenopausal and postmenopausal women. Perimenopausal and postmenopausal women with sleep disturbance should take sleep medication considerately, actively improve sleep quality, maintain good daytime function, andavoid or reduce cognitive impairment caused by sleep problems.

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    Effect of embryonic age on the pregnancy outcomes of blastocysts graded as CC in single frozen-thawed blastocyst transfer cycles
    Ming ZHAO, Ya HUANG, Tong DU, Jia-ying LIN, Xiu-juan QI, Li XU, Wen-yi LU, Yan-ping KUANG
    2021, 41 (4):  502-508. 
    doi: 10.3969/j.issn.1674-8115.2021.04.014

    Abstract ( 213 )   HTML ( 1 )   PDF (1179KB) ( 110 )  
    Objective

    · To assess the effect of embryonic age on the pregnancy outcomes of blastocyst graded as CC in single frozen-thawed blastocyst transfer cycles.

    Methods

    · A total of 565 patients undergoing single frozen-thawed blastocyst transfer at Shanghai TOWAKO Hospital from April 2017 to December 2019 were included in this study, and they were divided into high-quality blastocyst group (grade AA, AB, and BA; n=189), medium-quality blastocyst group (grade BB, AC, and CA; n=114), low-quality blastocyst group (grade CB and BC; n=140) and CC grade blastocyst group (n=122). And the CC grade blastocysts were further divided into CC grade blastocysts of Day 5 (D5 subgroup, n=26), Day 6 (D6 subgroup, n=81) and Day 7 (D7 subgroup, n=15) according to the embryonic age. Thereafter, the cycle characteristics and pregnancy outcomes of the four groups and the three embryonic age subgroups of CC grade blastocysts were compared.

    Results

    · The clinical pregnancy rate (58.7% vs 54.4% vs 47.1% vs 25.4%, P=0.000) and live birth rate (46.2% vs 43.2% vs 36.2% vs 17.6%, P=0.000) of the high-quality, medium-quality, low-quality, and CC grade blastocyst groups decreased sequentially. Multivariate analysis showed that embryo quality was significantly correlated with clinical pregnancy rate. Compared with high-quality blastocyst group, CC grade blastocyst group had a lower clinical pregnancy rate (aOR=0.81, 95%CI 0.71?0.93, P=0.003). The analysis of CC grade blastocyst embryonic age subgroups showed that the clinical pregnancy rate (42.3% vs 23.5% vs 6.7%, P=0.040) and live birth rate (31.6% vs 15.6% vs 0, P=0.133) of the D5, D6 and D7 subgroups also decreased sequentially. Multivariate analysis showed that the embryonic age had a significant impact on the clinical pregnancy rate of CC grade blastocyst transfer. Compared with the D5 subgroup, the D7 subgroup had a lower clinical pregnancy rate (aOR=0.69, 95%CI 0.49?0.96, P=0.032).

    Conclusion

    · D5 CC grade blastocysts predict good pregnancy outcome. Although the clinical pregnancy rate of D6 CC grade blastocysts is low, they are still useable. However, D7 CC grade blastocysts are of little clinical value.

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    Impact of intraoperative continuous pumping of remifentanil on postoperative analgesia after gastrointestinal surgery
    Lu-hong LI, Yan LUO
    2021, 41 (4):  509-513. 
    doi: 10.3969/j.issn.1674-8115.2021.04.015

    Abstract ( 220 )   HTML ( 1 )   PDF (1083KB) ( 152 )  
    Objective

    · To explore the effect of intraoperative continuous remifentanil infusion at the 0.05?0.15 μg/(kg·min) dosage on postoperative analgesia after gastrointestinal surgery.

    Methods

    · The patients received patient-controlled intravenous analgesia (PCIA) after elective gastrointestinal surgery from April to December 2019 in Ruijin Hospital, Shanghai Jiao Tong University School of Medicine were included in this retrospective analysis. They were divided into remifentanil group (n=360) and control group (n=335) according to whether the patients were treated with intraoperative continuous pumping of remifentanil at the 0.05?0.15 μg/(kg·min) dosage. Propensity score matching (PSM) was used to compare the difference of postoperative analgesic remedy and the pressing of patient-controlled analgesia (PCA) between the two groups.

    Results

    · By using a 1∶1 PSM analysis, 286 couples of patients were matched successfully. In the remifentanil group, the proportion of patients receiving postoperative analgesic remedies was significantly less than that in the control group (32.5% vs 40.9%, P=0.037). Compared with the control group, the remifentanil group had a lower risk of poor analgesic effect and necessity of postoperative analgesic remedies (RR=0.696, 95%CI 0.495?0.980). There was no significant difference in the total PCA pressing times between the remifentanil group and control group [5 (1, 12) vs 5 (1, 13), P=0.568], but in the remifentanil group, the proportion of patients with PCA pressing times > 13 was less than that of the control group (21.3% vs 24.5%).

    Conclusion

    · Continuous pumping of remifentanil at the 0.05?0.15 μg/(kg·min) dosage in elective gastrointestinal surgery is helpful to decrease postoperative analgesic demand. The patients receiving intraoperative remifentanil infusion at a low dosage has a lower risk in requiring postoperative analgesic remedies, compared with patients who do not receive intraoperative remifentanil infusion.

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    Clinical analysis of Kaposi form hemangioendothelioma in 19 children
    Jin-jun CAO, Wei XIE, Xiao-dong ZHU, Hao XIA
    2021, 41 (4):  514-518. 
    doi: 10.3969/j.issn.1674-8115.2021.04.016

    Abstract ( 198 )   HTML ( 1 )   PDF (1675KB) ( 125 )  
    Objective

    · To assess the clinical characteristics and treatment strategies of Kaposi form hemangioendothelioma (KHE) in children.

    Methods

    · The retrospective study was carried out on 19 cases of children diagnosed with KHE from January 2009 to December 2017 in Xinhua Hospital, Shanghai Jiao Tong University School of Medicine.The general demographics, tumor characteristics, presence or absence of Kasabach-Merritt phenomenon (KMP), treatment and outcome were statistically analyzed. The difference between the children with KMP and without KMP were compared in whether the tumor involved the skin and the deep muscles.

    Results

    · Of the 19 children with KHE, 13 (68.4%) were male and 6 (31.6%) were female; 17 children (89.5%) presented within the first year of life and 12 children (63.2%) were detected at birth; 6 cases occurred on head and neck, 4 cases in the trunk, 9 cases in the extremities, and none was multifocal.Seven cases (36.8%) were complicated with KMP after onset. There was no significant difference in cutaneous lesions (P=0.617), while there was a significant difference in deep muscles involvement (P=0.017) between the cases complicated with KMP and without KMP. All cases were diagnosed on the basis of histopathology and immunohistochemistry after operation, in which 8 were completely removed, 5 were partially excised, and 6 were performed biopsy. All the tumors were substantial and aggressive growth. Seven children were treated by propranolol before operation but no obvious effect was found. Ten children were given oral corticosteroids, and five of them were found tumor shrinking and underwent operation. The treatment of corticosteroids resulted in higher effect than that of propranolol (P=0.044). Two children who showed resistance to corticosteroids accepted a combination therapy of corticosteroids and vincristine, and the tumor shrank at last. During the follow-up of 2 to 5 years, the cases which performed complete resection were without recurrence, and those who accepted partial excision also survived with postoperative corticosteroids treatment.

    Conclusion

    · Most KHE cases occur in infancy. The cases might be predisposed with KPM when the tumors involve the deep muscles. The children with typical clinical features should undergo operation as soon as possible. Besides, those with atypical characters should take biopsy to achieve definite diagnosis. The cases with KMP or with giant tumors that are impossible to be removed could be given corticosteroids treatment with or without vincristine. Although propranolol showed a poor curative effect, it could be used for a diagnostic treatment.

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    Public health
    Mediation effects of energy balance-related behaviors on associations between sleep and the risk of obesity among elementary and junior high school students
    Shu-ping LOU, Yan HUANG, Pan-yue LIU, Hui CHEN
    2021, 41 (4):  519-524. 
    doi: 10.3969/j.issn.1674-8115.2021.04.017

    Abstract ( 257 )   HTML ( 2 )   PDF (1272KB) ( 125 )  
    Objective

    · To investigate the possible mediation effects of energy balance-related behaviors on the association of sleep with obesity among elementary and junior high school students.

    Methods

    · Data of 5 179 primary and middle school students were assessed via a self-reported questionnaire and anthropometric measurement, including sleep variables, physical activities, screen behaviors, dietary patterns, height and weight status in Maoming City, Guangdong Province from September to November 2016. The enrolled students were divided into obese group and non-obese group according to body mass index threshold of Screening of Overweight and Obesity in School-age Children and Adolescents in China. Linear regression and Logistic regression analyses were used to estimate the associations between sleep, the aforementioned energy balance-related behaviors and obesity. The MacKinnon's product-of-coefficients method was used to assess the mediation effects of energy balance-related behaviors.

    Results

    · Compared with the non-obese group, most obese students were characterized by being male, young, parental obesity, high monthly house income, long screen time, high frequency of high calorie food and fruit-milk diet. Significant associations were observed between sleep duration, bedtime and wake-up time of elementary and junior high school students with the risk of obesity (all P=0.000). The intermediary pathways of obesity (sleep→screen behavior→obesity, and sleep→physical activity→obesity) were statistically significant (P<0.05). No evidence for the mediating role of high calorie or fruit-milk dietary habits was found in this study.

    Conclusion

    · Physical activity and screen behavior were identified as possible mediators of the association between sleep and obesity.

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    Review
    Advances in epidemiology and treatment of invasive Candida infection in the immunocompromised population
    Lin LIU, Hui WANG, Ning-ning LIU
    2021, 41 (4):  525-529. 
    doi: 10.3969/j.issn.1674-8115.2021.04.018

    Abstract ( 278 )   HTML ( 3 )   PDF (975KB) ( 133 )  

    With the development of modern medical technology, such as organ transplantation, the extensive application of broad-spectrum antibiotics and immunosuppressants and the prolongation of human life span, the number of people with low immunity is increasing. The morbidity and mortality of invasive fungal infection are increasing in these population. Invasive fungal infection has become the direct cause of death of critical diseases, such as malignant tumor and organ transplantation. Candida is the primary pathogenic fungi of invasive fungal infection. It can infect human subcutaneous tissue, mucous membrane, other tissues and organs of the whole body, and then spread in the blood. The main clinical manifestations of Candida infectionare candidemia and deep candidiasis, and the prognosis is often poor. At present, antifungal infection is still the main treatment for invasive Candida (IC) in immunocompromised population, and drug treatment is one of the effective ways. Clinical conventional antifungal drugs include polyenes, azoles and echinocandins. Among them, echinocandins is the first choice for the treatment of IC. However, the existing treatments fail to improve the prognosis of patients with IC infection, and drug resistance caused by long-term use frequently occurr. At present, the number of antifungal drugs available is extremely limited. Therefore, in order to solve the above problems, drug combination therapy, adjuvant immunotherapy, photodynamic therapy and laser therapy provide new perspectives for the treatment of IC. This article reviews the epidemiology and treatment of IC infetcion in the population with low immunity. In this paper, the incidence and mortality of IC infection in patients in intensive care unit, patients with hematological malignancies, solid organ transplant patients, neonates, the elderly, and human immunodeficiency virus (HIV)-infected people at home and abroad, and the distribution of main pathogenic Candida species, as well as the therapeutic potential and limitations of new research were discussed in depth. The future research direction was proposed.

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    Review and prospect of anti-vascular endothelial growth factor treatment for wet age-related macular degeneration
    Ya-fang WANG, Yang LIU, Xue-ting LUO
    2021, 41 (4):  530-534. 
    doi: 10.3969/j.issn.1674-8115.2021.04.019

    Abstract ( 255 )   HTML ( 6 )   PDF (1129KB) ( 141 )  

    Anti-vascular endothelial growth factor (VEGF) has been the first choice in the treatment of wet age-related macular degeneration (AMD), since ranibizumab went on sale in 2006. However, clinical studies have suggested that the visual acuity returned to a baseline level several years after anti-VEGF therapy, with the occurrence of retinal fibrosis and geomorphologic atrophy, highlighting the limit of this treatment. In order to improve the efficacy, replacement and supplementation of traditional anti-VEGF emerged successively. Gene therapy achieved long-term and stable anti-VEGF effect by mediating therapeutic proteins delivered by adeno-associated virus vectors. Treatment combined with anti-platelet-derived growth factor may compensate for the limitations of anti-VEGF therapy through peripheral cells. Complement-related gene therapy is a new field based on the poor reactivity of patients against VEGF. This article reviews the progress of anti-VEGF in the treatment of wet AMD.

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    Progress of catabolite control protein A in the regulation of metabolism and virulence of Gram-positive bacteria
    Zi-yu YANG, Juan-xiu QIN, Min LI, Qian LIU
    2021, 41 (4):  535-539. 
    doi: 10.3969/j.issn.1674-8115.2021.04.020

    Abstract ( 226 )   HTML ( 4 )   PDF (1076KB) ( 120 )  

    Catabolite control protein A (CcpA) is an important pleiotropic regulator in Gram-positive bacteria. With the assistance of the phosphoenolpyruvate-dependent carbohydrate phosphotransferase system, CcpA undergoes a conformational change and interacts with catabolite responsive element, a specific DNA sequence on the target gene, activating or repressing expression of the target gene, thereby controlling the physiological processes and virulence of bacteria. CcpA mediates the carbon catabolite repression effect of microorganisms to control the uptake and utilization of environmental nutrients, ensuring the efficient use of energy by bacteria. CcpA regulates bacterial metabolism and virulence gene expression directly or indirectly, coupling the metabolism and virulence of bacteria. This review describes the mechanism of CcpA function, and summarizes the research progress of CcpA in regulating multiple metabolic pathways and pathogenicity of bacteria.

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    Advance in research of interference effect of perfluorinated compounds on sexhormones
    Zi-xia WANG, Qian YAO, Ying TIAN
    2021, 41 (4):  540-545. 
    doi: 10.3969/j.issn.1674-8115.2021.04.021

    Abstract ( 184 )   HTML ( 6 )   PDF (1151KB) ( 119 )  

    Perfluorinated compounds (PFCs) are a class of persistent organic pollutants that are widely distributed in the environment and biota. Studies have shown that PFCs have liver-toxicity, immune-toxicity and neuro-toxicity, etc. In recent years, due to an increasing number of studies suggesting that PFCs have endocrine disrupting effects, their reproductive toxicity has attracted much attention, especially their interference with the synthesis and secretion of sex hormone which may induce abnormalities in sexual differentiation and development, reproductive disorders, etc.This article reviews the relationship of PFCs with sex hormone levels and the potential mechanisms from animal toxicology experiments and epidemiological studies, thus providing the theoretical basis for PFCs research in the future.

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    Research progress of children's pain memories
    Feng-qiao WANG, Lei ZHUANG, Fu-jun ZHANG
    2021, 41 (4):  546-549. 
    doi: 10.3969/j.issn.1674-8115.2021.04.022

    Abstract ( 235 )   HTML ( 3 )   PDF (898KB) ( 96 )  

    In recent years, people have discovered painful memories in infancy. Children's memory of pain is plastic and related to pain trajectory and chronic pain. It is an integral part of pain assessment and treatment. Based on the results of recent clinical studies, this article reviews the definition, plasticity, clinical significance, influencing factors, and interventions of children's pain memory. The aim is to provide reference for the development of children's pain management, clinical practice and scientific research.

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    Research progress on preparation materials of anti-adhesion membranes in the prevention of tendon adhesion
    Ming-kuan LU, Chuan-dong CAI, Wei WANG, Cun-yi FAN, Shen LIU
    2021, 41 (4):  550-553. 
    doi: 10.3969/j.issn.1674-8115.2021.04.023

    Abstract ( 229 )   HTML ( 7 )   PDF (1003KB) ( 132 )  

    Adhesion formation after tendon repair is still a challenge in current clinical practice. With the development of material science, more and more materials are used to prepare anti-adhesion membranes for the prevention and treatment of tendon adhesion. Various materials have different physical and chemical features, different localreactions and curative effects at the injury site, so they have their own advantages and disadvantages. This article highlights the common natural biomaterials and synthetic materials used to prepare anti-adhesion membranes, as well as the basic research and clinical applications of anti-adhesion membranes based on various materials, in order to play a reference role in optimizing the design and construction of anti-adhesion membranes.

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    Research advances in stem cell-based therapies for acute respiratory distress syndrome
    Jia-yang YAN, Yun FENG, Jie-ming QU
    2021, 41 (4):  554-558. 
    doi: 10.3969/j.issn.1674-8115.2021.04.024

    Abstract ( 179 )   HTML ( 4 )   PDF (1050KB) ( 163 )  

    Acute respiratory distress syndrome (ARDS) is caused by various factors which damage pulmonary alveolar capillary barrier. The morbidity and mortality are high, but there are no effective and specific treatments. A large number of studies have identified that stem cells can ameliorate lung injury in various ways. As novel biological therapy methods, stem cell-based therapies have achieved good results in ARDS preclinical models while clinical trials have shown that the therapeutic efficacy is unsatisfactory. Accordingly, several strategies have been explored to strengthen the therapeutic potential of stem cells. This article reviews research advances in stem cell-based therapies for ARDS and strategies to enhance the therapeutic potential of stem cells.

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