1.Cloning and functional analysis of GmMAX2b involved in disease resistance.
Jiahui FU ; Lin ZUO ; Weiqun HUANG ; Song SUN ; Liangyu GUO ; Min HU ; Peilan LU ; Shanshan LIN ; Kangjing LIANG ; Xinli SUN ; Qi JIA
Chinese Journal of Biotechnology 2025;41(7):2803-2817
The plant F-box protein more axillary growth 2 (MAX2) is a key factor in the signal transduction of strigolactones (SLs) and karrinkins (KARs). As the main component of the SKP1-CUL1-FBX (SCF) complex ubiquitin ligase E3, MAX2 is responsible for specifically recognizing the target proteins, suppressor of MAX2 1/SMAX1-like proteins (SMAX1/SMXLs), which would be degraded after ubiquitination. It can thereby regulate plant morphogenesis and stress responses. There exist homologous genes of MAX2 in the important grain and oil crop soybean (Glycine max). However, its role in plant defense responses has not been investigated yet. Here, GmMAX2b, a homologous gene of MAX2, was successfully cloned from stressed soybean. Bioinformatics analysis revealed that there were two MAX2 homologous genes, GmMAX2a and GmMAX2b, with a similarity of 96.2% in soybean. Their F-box regions were highly conserved. The sequence alignment and cluster analysis of plant MAX2 homologous proteins basically reflected the evolutionary relationship of plants and also suggested that soybean MAX2 might be a multifunctional protein. Expression analysis showed that plant pathogen infection and salicylic acid treatment induced the expression of GmMAX2b in soybean, which is consistent with that of MAX2 in Arabidopsis. Ectopic expression of GmMAX2b compensated for the susceptibility of Arabidopsis max2-2 mutant to pathogen, indicating that GmMAX2b positively regulated plant disease resistance. In addition, yeast two hybrid technology was used to explore the potential target proteins of GmMAX2b. The results showed that GmMAX2b interacted with SMXL6 and weakly interacted with SMXL2. In summary, GmMAX2b is a positive regulator in plant defense responses, and its expression is induced by pathogen infection and salicylic acid treatment. GmMAX2b might exert its effect through interaction with SMXL6 and SMXL2. This study expands the theoretical exploration of soybean disease resistant F-box and provides a scientific basis for future soybean disease resistant breeding.
Glycine max/metabolism*
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Disease Resistance/genetics*
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Plant Diseases/immunology*
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Plant Proteins/genetics*
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Cloning, Molecular
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Gene Expression Regulation, Plant
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F-Box Proteins/genetics*
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Arabidopsis/genetics*
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Phylogeny
2.PGRMC1-mediated autophagy decreases the sensitivity of hepatocellular carcinoma cells to 125I particle irradiation
Pingping LIU ; Chenyu WANG ; Yunhua XIAO ; Chuang HE ; Junru XIONG ; Liangyu DENG ; Xuequan HUANG
Journal of Army Medical University 2024;46(9):1015-1023
Objective To investigate the effect of progesterone receptor membrane component 1(PGRMC1)mediated autophagy on the sensitivity of liver cancer cells to 125I particles irradiation.Methods Hepatoma cell lines Huh7 and LM3 were exposed to different doses(0,2,4,6 and 8 Gy)of 125I particles,and cell autophagy was observed by transmission electron microscopy(TEM).Then,autophagy inhibitor chloroquine(CQ),agonist rapamycin(Rapa),and PGRMC1 inhibitor AG-205 were used respectively to verify that PGRMC1-mediated autophagy plays a key role in the sensitivity of hepatocellular carcinoma cells to 125I particle irradiation.Cell proliferation,colony formation and apoptosis were detected by CCK-8 assay,clonal formation test and flow cytometry,respectively.The expression levels of PGRMC1,microtubule-associated protein light chain 3-Ⅰ(LC3-Ⅰ),LC3-Ⅱ and p62 were detected by Western blotting.Results Different doses of 125I particles irradiation significantly decreased the proliferation and clonogenesis of Huh7 and LM3 cells(P<0.05),and increased the apoptotic cells(P<0.01),in a dose-dependent manner.Compared with the 0 Gy group,the ratio of LC3-Ⅱ/LC3-Ⅰ in Huh7 and LM3 cells was obviously increased,and the expression of p62 was significantly down-regulated in the 6 Gy group.The proliferation capacity and clonal formation ability of Huh7 and LM3 cells were decreased significantly,and their apoptotic cells were increased notably in the 6 Gy+CQ group than the 6 Gy group,while the above results were on the contrary in the 6 Gy+Rapa group.The 6 Gy+AG205 group had notably decreased LC3-Ⅱ/LC3-Ⅰ ratio in the Huh7 and LM3 cells,up-regulated p62 expression,reduced cell proliferation capacity and clone formation ability,and enhanced cell apoptosis when compared with the 6 Gy group,and the above results of the 6 Gy+PGRMC1 group were opposite.Conclusion Increment of PGRMC1 induced by 125I irradiation can promote autophagy,increase the proliferation and clonogenesis,and reduce the apoptosis in hepatocellular carcinoma cells.
3.Status quo of psychological contracts among members of the“1+N”family doctor teams in Shenzhen and the impact on job burnout
Shuangshuang WEN ; Yue DU ; Mulan JIANG ; Liangyu HUANG ; Qihua MO ; Ye LIU ; Jianwei ZHANG ; Hegao YU ; Liang MA
Chinese Journal of General Practitioners 2024;23(8):848-854
Objective:To investigate the status quo of psychological contracts and influencing factors among members of the "1+N" family doctor teams in Shenzhen and to explore the impact of psychological contracts on job burnout.Methods:This cross-sectional study was conducted from September 30 to October 31, 2022 among 361 members of 92 family doctor teams from 92 community health service centers which provided family doctor team service in Shenzhen city. A self-designed general information questionnaire, an employee psychological contract questionnaire (including organizational responsibility and personal responsibility dimensions), and a job burnout scale (including emotional exhaustion, depersonalization, and personal accomplishment dimensions) were used in the study. T-tests, one-way ANOVA, Pearson correlation analysis, and multiple linear regression analysis were used to analyze the influencing factors of psychological contracts and job burnout.Results:Among 361 respondents, there were 299 females (82.8%) and 62 males (17.2%), and a higher proportion of general practitioners (37.5%, 129/361) and nurses (41.8%, 151/361). The total score of psychological contracts among the 361 respondents was (141.6±19.5), with organizational responsibility scoring (70.6±11.2) and personal responsibility scoring (71.0±9.3). On the job burnout scale, emotional exhaustion scored (17.89±6.82), depersonalization scored (6.51±2.54), and personal accomplishment scored (30.95±5.70). General practitioners scored lower in organizational responsibility and personal responsibility compared to other members ( F=7.341,3.119, all P<0.05), and higher in emotional exhaustion and depersonalization ( F=7.637, 2.415, all P<0.05). Members with≤5 years of work experience scored lower in personal responsibility and personal accomplishment ( F=3.656, 4.205, all P<0.05). Correlation analysis showed that scores of organizational responsibility and personal responsibility were negatively correlated with levels of emotional exhaustion and depersonalization ( r=-0.618, -0.526, all P<0.01), ( r=-0.404, -0.393, all P<0.01), and positively correlated with personal accomplishment ( r=0.500, 0.558, all P<0.01). Multiple linear regression analysis indicated that organizational responsibility negatively affected emotional exhaustion and depersonalization ( β=-0.554, -0.274, all P<0.01), and positively affected personal accomplishment ( β=0.172, P<0.05). Personal responsibility positively affected personal accomplishment ( β=0.404, P<0.01). Conclusions:The study demonstrates that general practitioners in family doctor teams in Shenzhen city have lower psychological contract levels and are more prone to emotional exhaustion and depersonalization; members with≤5 years of work experience have lower personal responsibility and accomplishment. The results indicate that enhancing organizational responsibility can reduce job burnout of members in family doctor teams.
4.A case of iatrogenic hypernatremia caused by multiple myeloma-associated pseudohyponatremia
Yuan HUANG ; Lu ZHANG ; Liangyu XIA ; Jinyan LEI ; Chong WEI ; Yongzhe LI
Chinese Journal of Laboratory Medicine 2024;47(10):1212-1214
The patient was a 55-year-old man. On February 16, 2024, he was admitted to Peking Union Medical College Hospital complaining of "weakness and poor appetite for more than half a year, and found creatinine increase for 1 week". The patient was diagnosed with multiple myeloma. During the treatment in our hospital, the patient sustained"hyponatremia"(Na 124-136 mmol/L measured by indirect ion selective electrode method), and combined with the patient′s clinical symptoms and serum osmotic pressure results (327 mOsm/kg H 2O), it was considered that hyperglobulinemia led to pseudohyponatremia. So no intervention was given. Subsequent failure to recognize pseudohyponatremia during treatment in other hospitals and the administration of hypertonic saline resulted in severe iatrogenic hypernatremia. By reviewing similar cases in our hospital, we found that hyperglobulinemia/hyperlipidemia associated pseudohyponatremia was not uncommon. This case reminds us that for patients whose serum solid phase ratio is higher than normal due to various reasons, the use of indirect ion selective electrode method to determine serum sodium is prone to false low, and direct ion selective electrode method can be used to re-test blood sodium to determine whether it is true, so as to avoid iatrogenic injury to patients.
5.Multi-slice spiral CT multi-planar reconstruction technology applicated in CT-guided puncture biopsy of pancreatic lesions
Liangyu DENG ; Xuequan HUANG ; Yunhua XIAO ; Chuang HE
Chinese Journal of Interventional Imaging and Therapy 2024;21(12):781-784
Objective To explore the application value of multi-slice spiral CT(MSCT)multi-planar reconstruction(MPR)technology for CT-guided puncture biopsy of pancreatic lesions.Methods A total of 104 patients with isolated pancreatic lesions who underwent CT-guided puncture biopsy were retrospectively enrolled,including 49 cases underwent MSCT-MPR assisted guided biopsy(MPR group)and 55 underwent MSCT-guided biopsy alone(non-MPR group).The success rate of puncture biopsy,the operating time,the times of needle adjustments,radiation dose-length product(DLP)and relative complications were recorded.Based on surgical pathology or follow-up(≥12 months)results,the sensitivity of biopsy was calculated.Results The success rate of one-time puncture was 100%in both groups.No significant difference of operating time was found between MPR([15.96±4.73]min)and non-MPR group([16.22±5.76]min,t=-0.250,P=0.803).MPR group had fewer times of needle adjustments([6.53±1.24]vs.[8.15±1.90],t=-5.201,P<0.001)and lower DLP([946.02±143.54]mGy·cm vs.[1 095.53±338.50]mGy·cm,t=-2.988,P=0.004)than non-MPR group.During biopsy,minor bleeding occurred in 1 case of MPR group and 5 of non-MPR group,then all spontaneously improved without special treatment.The diagnostic sensitivity of biopsy pathology was 95.56%(43/45)in MPR group and 92.16%(47/51)in non-MPR group.Conclusion MSCT-MPR technology could improve the accuracy of CT-guided puncture biopsy of pancreatic lesions and reduce radiation dose.
6.Multi-slice spiral CT multi-planar reconstruction technology applicated in CT-guided puncture biopsy of pancreatic lesions
Liangyu DENG ; Xuequan HUANG ; Yunhua XIAO ; Chuang HE
Chinese Journal of Interventional Imaging and Therapy 2024;21(12):781-784
Objective To explore the application value of multi-slice spiral CT(MSCT)multi-planar reconstruction(MPR)technology for CT-guided puncture biopsy of pancreatic lesions.Methods A total of 104 patients with isolated pancreatic lesions who underwent CT-guided puncture biopsy were retrospectively enrolled,including 49 cases underwent MSCT-MPR assisted guided biopsy(MPR group)and 55 underwent MSCT-guided biopsy alone(non-MPR group).The success rate of puncture biopsy,the operating time,the times of needle adjustments,radiation dose-length product(DLP)and relative complications were recorded.Based on surgical pathology or follow-up(≥12 months)results,the sensitivity of biopsy was calculated.Results The success rate of one-time puncture was 100%in both groups.No significant difference of operating time was found between MPR([15.96±4.73]min)and non-MPR group([16.22±5.76]min,t=-0.250,P=0.803).MPR group had fewer times of needle adjustments([6.53±1.24]vs.[8.15±1.90],t=-5.201,P<0.001)and lower DLP([946.02±143.54]mGy·cm vs.[1 095.53±338.50]mGy·cm,t=-2.988,P=0.004)than non-MPR group.During biopsy,minor bleeding occurred in 1 case of MPR group and 5 of non-MPR group,then all spontaneously improved without special treatment.The diagnostic sensitivity of biopsy pathology was 95.56%(43/45)in MPR group and 92.16%(47/51)in non-MPR group.Conclusion MSCT-MPR technology could improve the accuracy of CT-guided puncture biopsy of pancreatic lesions and reduce radiation dose.
7.Intelligent identification of livestock, a source of Schistosoma japonicum infection, based on deep learning of unmanned aerial vehicle images
Jingbo XUE ; Shang XIA ; Zhaojun LI ; Xinyi WANG ; Liangyu HUANG ; Runchao HE ; Shizhu LI
Chinese Journal of Schistosomiasis Control 2023;35(2):121-127
Objective To develop an intelligent recognition model based on deep learning algorithms of unmanned aerial vehicle (UAV) images, and to preliminarily explore the value of this model for remote identification, monitoring and management of cattle, a source of Schistosoma japonicum infection. Methods Oncomelania hupensis snail-infested marshlands around the Poyang Lake area were selected as the study area. Image datasets of the study area were captured by aerial photography with UAV and subjected to augmentation. Cattle in the sample database were annotated with the annotation software VGG Image Annotator to create the morphological recognition labels for cattle. A model was created for intelligent recognition of livestock based on deep learning-based Mask R-convolutional neural network (CNN) algorithms. The performance of the model for cattle recognition was evaluated with accuracy, precision, recall, F1 score and mean precision. Results A total of 200 original UAV images were obtained, and 410 images were yielded following data augmentation. A total of 2 860 training samples of cattle recognition were labeled. The created deep learning-based Mask R-CNN model converged following 200 iterations, with an accuracy of 88.01%, precision of 92.33%, recall of 94.06%, F1 score of 93.19%, and mean precision of 92.27%, and the model was effective to detect and segment the morphological features of cattle. Conclusion The deep learning-based Mask R-CNN model is highly accurate for recognition of cattle based on UAV images, which is feasible for remote intelligent recognition, monitoring, and management of the source of S. japonicum infection.
8.Chitinase-3-like protein 1 plays a role in lumbar facet joint degeneration
Yuanzhen CHEN ; Lei WANG ; Liangyu XIE ; Shengnan CAO ; Xiuchun YU ; Huazhong LI ; Weimin HUANG ; Bin SHI ; Dandan WANG
Chinese Journal of Orthopaedics 2022;42(17):1148-1155
Objective:To investigate the effect of CHI3L1 on the biological function of chondrocytes and its role in lumbar facet joint degeneration.Methods:The human lumbar facet joint articular cartilage were collected, and the relative mRNA expression of CHI3L1 gene detected by quantitative fluorescence PCR. Then explored the correlation between joint degeneration and gender, age and relative mRNA expression of CHI3L1. Human chondrocytes were cultured in vitro. The effects of CHI3L1 on chondrocyte proliferation, cycling, and apoptosis, as well as expression of related inflammatory factors, were investigated. The mechanism by which CHI3L1 regulates the degeneration of articular cartilage was investigated using the signal transduction pathway protein chip.Results:There was a positive correlation between the grade of degeneration in lumbar facet joint and the relative expression of CHI3L1 gene mRNA ( r=0.76, P<0.001). There was no correlation with the patient's gender ( r=-0.12, P=0.500). A positive correlation between the age of patients and the relative expression of CHI3L1 gene mRNA was found ( r=0.47, P=0.005). Compared with the non-degenerative group, the expression of CHI3L1 gene mRNA significantly increased in the degenerative group, and the expression of CHI3L1 gradually increased with the aggravation in the grade of degeneration ( F=18.90, P<0.001). Compared with the non-degenerative group, the chondrocytes in the CHI3L1 group had significantly lower proliferation at 48 h (OD 490/fold=7.132), 72 h (OD 490/fold=4.803), 96 h (OD 490/fold=2.431) and 120 h (OD 490/fold=0.009). The ratio of chondrocytes in G1 phase, S phase and G2/M phase were 85.03%±3.05%, 12.78%±2.29% and 0.90%±0.76% in the CHI3L1 group, and 73.93%±2.73%, 22.81%±1.93% and 0.99%±0.87% in control group, respectively. There were significant differences in the percentage of chondrocytes in G1 phase ( t=4.70, P<0.001) and S phase ( t=5.80, P<0.001) between the two groups. The percentages of apoptosis in chondrocyte in CHI3L1 group and control group were 8.64%±0.76% and 5.68%±1.13%, which has a statistically difference ( t=4.47, P<0.001). The expression of IL-6 in chondrocytes of CHI3L1 group was 49.60±0.01 pg/ml, which was higher than that of 47.88±0.01 pg/ml in the control group ( t=132.70, P<0.001). The expression of TNF-α was 95.93±0.02 pg/ml, which was higher than 90.69±0.02 pg/ml in the control group ( t=376.10, P<0.001). There was significant difference in expression of IL-6 in chondrocytes between the CHI3L1 group and the control group ( t=132.72, P<0.001). The expression of TNF-α ( t=376.10, P<0.001) was statistically difference. Protein chip detected 53 proteins with significant differences in expression and 43 proteins with significant differences in protein phosphorylation levels. Bioinformatics analysis was used to identify 16 signaling pathways in which the above different proteins might be involved, including ErbB, PI3K, Akt, Ras, JAK, STAT3, MAPK pathway. In the MAPK pathway, the expression of MAPK1 and RAF1 proteins was higher in the chondrocytes of the CHI3L1 group than in the control group (1.094±0.00 vs. 0.814±0.00, 0.988±0.00 vs. 0.786±0.00; t=103.16, P<0.001; t=54.32, P<0.001). Compared with the control group, the expression of MAPK1 and RAF1 proteins was significantly increased in the chondrocytes of the CHI3L1 group. Conclusion:The expression of CHI3L1 is corrected to articular cartilage degeneration. CHI3L1 is able to inhibit the proliferation of articular chondrocytes, which regulated the cycling of chondrocytes from G1 phase to S phase, promote the apoptosis of chondrocytes, and promote the expression of IL-6 and TNF-α in chondrocytes. Regulation of chondrocytes biological function through the MAPK pathway, which is a potential biomarker for the clinical diagnosis and treatment of lumbar joint degeneration.
9.Maternal iron status in the first trimester is associated with gestational diabetes mellitus and macrosomia
Feiling HUANG ; Liangkun MA ; Shuai MA ; Yixuan HOU ; Aimin YAO ; Liangyu XIA ; Songlin YU ; Shanshan LI ; Haoze LUO ; Zikun ZHOU ; Jiaxin ZHANG ; Yanping LIU
Chinese Journal of Health Management 2020;14(6):515-520
Objective:To evaluate the association of maternal nutrition status in the first trimester with gestational diabetes mellitus (GDM) and macrosomia.Methods:378 pregnant women who took prenatal care in Shunyi Women′s and Children′s Hospital of Beijing Children′s Hospital were enrolled in the study. Blood samples were collected at first prenatal visit (<12 gestation weeks) to measure the level of hemoglobin and iron status indexes including serum iron, ferritin, transferrin, total iron binding capacity, iron saturation, transferrin saturation. The incidence of GDM and macrosomia were collected and Logistic regression was used to evaluate the associations of maternal nutrients status in the first trimester with GDM and macrosomia.Results:The incidence rate of GDM was16.9%,the incidence of anemia and iron deficiency in the first trimester were2.4% and 2.5%, respectively. After adjustment for variables such as maternal age, pre-pregnancy BMI, family history of diabetes, and parity, Logistic regression showed that in the first trimester, iron saturation>50% ( OR=0.238, 95% CI 0.068-0.831), transferrin saturation>50% ( OR=0.08, 95% CI 0.010-0.677) were protective factors of GDM; iron saturation 25%-50% ( OR=0.361, 95% CI 0.143-0.908); transferrin saturation 25%-50% ( OR=0.383, 95% CI 0.165-0.891); ferritin>30 ng/ml ( OR=0.418, 95% CI0.186-0.939) were protective factors of macrosomia. Conclusion:Maternal iron status in the first trimester might be associated with GDM and macrosomia. Thus, maternal iron status assessment in the first trimester is necessary.
10.Protective effect of polypyrimidine bundle-binding protein-related splicing factor on retinal pigment epithelial cell injury induced by advanced glycation end products
Chen QI ; Hui ZHANG ; Tingting LIN ; Yifeng KE ; Xinjun REN ; Shaochong BU ; Liangyu HUANG ; Yong WANG ; Mingfei JIAO ; Liying HU ; Qiong WANG ; Yaru HONG ; Xiaorong LI ; Lijie DONG
Chinese Journal of Ocular Fundus Diseases 2020;36(1):46-52
Objective To observe the protective effect of polypyrimidine bundle-binding proteinrelated splicing factor (PSF) over-expression on RPE cell injury induced by advanced glycation end products (AGEs).Methods The human RPE cells cultured in vitro were divided into three groups:normal control group (N group),blank control group (N + AGEs group),empty vector control group (Vec + AGEs group),and PSF high expression group (PSF + AGEs).group).RPE cells in N group were routinely cultured;RPE cells in N + AGEs group were only transfected but did not introduce any exogenous genes combined with AGEs induction;Vec +AGEs group and PSF + AGEs group were transfected with pcDNA The empty vector or pcDNA-PSF eukaryotic expression plasmid was introduced into RPE cells and induced by AGEs.Except the N group,the other 3 groups of cells were transfected accordingly,and were stimulated with 150 μg/ml AGEs for 72 h after 24 h.HE staining and Hoechst 33258 staining were used to observe the effect of high PSF expression on the morphological changes of RPE cells;ROS level detection was used to analyze the effect of PSF high expression on the ROS expression of RPE cells induced by AGEs;MTT colorimetric method was used to detect the high PSF expression Effects on the viability of RPE cells;Western blot was used to detect the effects of different time and dose of PSF on the expression of heme oxygenase 1 (HO-1).Results HE staining and Hoechst 33258 staining observation showed that the cells in group N were full in shape,the nucleus was round,the cytoplasm was rich,and the staining was uniform;the cells in N + AGEs group and Vec + AGEs group were reduced in size,the eosinophilic staining was enhanced,and the nucleus was densely densely stained.Pyrolysis and even fragmentation;the morphology of cells in the PSF + AGEs group was still full,the cytoplasm staining was more uniform,and the nucleus staining was uniform.The results of MTT colorimetry showed that high expression of PSF can effectively improve the viability of RPE cells,but this effect can be effectively antagonized by ZnPP,and the difference is statistically significant (F=33.26,P<0.05).DCFH-DA test results showed that compared with the N + AGEs group and Vec + AGEs group,the ROS production in PSF + AGEs group decreased,the difference was statistically significant (F=1 1.94,P<0.05).Western blot analysis showed that PSF protein upregulated HO-1 expression in a time-and dose-dependent manner.The relative expression level of HO-1 at 24,48,and 72 h after PSF protein was significantly higher than that at 0 h,and the difference was statistically significant (F=164.91,P<0.05).The relative expression level of HO-1 under the action of 0.1,0.5,1.0,1.5,and 2.0 μg PSF protein was significantly higher than 0.0 μg,and the difference was statistically significant (F=104.82,P<0.05).Conclusion PSF may inhibit the production of ROS by up-regulating the expression of HO-1,thus protecting the RPE cells induced by AGEs.

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