1.Expression of TFDP2 in placentas of women with preeclampsia and its effect on apoptosis of trophoblast cells
Chenrui CAO ; Dan LIU ; Zhiyin WANG ; Guangfeng ZHAO ; Zhongrui PEI ; Yali HU
Chinese Journal of Perinatal Medicine 2024;27(2):133-142
Objective:To investigate the expression level of transcription factor dimerization partner 2 (TFDP2) in the placentas of women with preeclampsia, and analyze its effect on the apoptosis of trophoblast cells.Methods:Placental tissues from thirty puerperae with preeclampsia who gave birth by cesarean section in Nanjing Drum Tower Hospital, the Affiliated Hospital of Nanjing University Medical School between January 2018 and December 2022 (preeclampsia group) and 30 healthy puerperae undergoing cesarean section during the same period (control group) were retrospectively selected. Immunohistochemistry was used to localize TFDP2 in the placental tissues. Real-time quantitative-polymerase chain reaction (qRT-PCR) and Western blot were used to detect the differences in expression of TFDP2 at mRNA and protein levels in placental tissues between the two groups. Forskolin-exposed BeWo cells were transfected with small interfering RNA (siRNA) to knockdown TFDP2 and the changes in the expression of apoptosis-related indicators, B cell lymphoma 2 (Bcl2) and Bcl2 associated X (Bax), at protein and mRNA levels were analyzed by Western blot and qRT-PCR, respectively. Besides, the change in the apoptosis level of BeWo cells was detected using terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) staining and flow cytometry. Downstream signaling pathways were analyzed to understand the involved molecular mechanisms. Two independent samples t-test, Wilcoxon rank-sum test, and Chi-square test were used for statistical analysis. Results:TFDP2 was mostly localized in the syncytiotrophoblasts and the extravillous trophoblasts in the normal placentas. TFDP2 expression in the syncytiotrophoblasts was lower in the preeclampsia group than in the control group at both mRNA (0.722±0.239 vs. 1.000±0.348, t=3.61, P=0.001) and protein (0.728±0.185 vs. 1.000±0.206, t=2.41, P=0.037) levels. Comparing the group without knockdown of TFDP2, the knockdown of TFDP2 in BeWo cells elevated the Bax/Bcl2 ratio (mRNA: 1.755±0.452 vs. 1.000±0.279, t=3.48, P=0.006; protein: 3.206±0.922 vs. 1.000±0.290, t=3.95, P=0.017), and increased cell apoptosis both in number and ratio (TUNEL staining: 4.556±1.740 vs. 2.444±1.130, t=3.05, P=0.008; flow cytometry: 21.37%±1.66% vs. 12.61%±0.38%, t=8.92, P=0.001). Furthermore, following TFDP2 knockdown, a decrease in the phosphorylation activity of catalytic subunit of protein kinase A (PKAc) at the Thr197 site was observed in the cytoplasm of BeWo cells (0.466±0.035 vs. 1.000±0.075, t=11.19, P<0.001) and a reduction in the expression of β-catenin in the cell nucleus was also detected (0.250±0.093 vs. 1.000±0.269, t=4.57, P=0.010). Conclusion:The expression of TFDP2 decreased significantly in the placentas of patients with preeclampsia, which may promote the apoptosis of syncytiotrophoblasts by inhibiting the PKAc/β-catenin signaling pathway.
2.Application of laparoscopic single-incision triangulated umbilical surgery technique in urology
Yuhao YU ; Zhiguang ZHAO ; Gen LI ; Xuexing FAN ; Zhiguo LU ; Guangfeng ZHU ; Xiaoliang DOU ; Xiaopeng CHEN ; Bo ZHAO ; Yong WANG
Chinese Journal of Urology 2024;45(3):238-240
The pursuit of cosmetic effects in post-surgical wounds has led to the development of ultra-minimally invasive techniques in surgery. Minimal invasive surgery has replaced open surgery and has become the new gold-standard for treating diseases. One such technique is the single incision triangulated umbilicus surgery (SITUS), which offers several advantages over traditional laparoscopic and other scarless surgeries, including reduced trauma, faster recovery, and better cosmetic outcomes. SITUS also has a short learning curve, aligns with conventional instrumentation operating habits, and can be used for whole abdominal surgeries. Chinese scholars have made further improvements to the SITUS technology, including expanding its applicability in intra-abdominal surgery and refining its incision closure methods to achieve superior cosmetic results. Currently, SITUS technology is experiencing rapid development in urology applications and has demonstrated satisfactory results in both domestic and international reports. This review aims to discuss the effectiveness and development of the SITUS technique in urology.
3.Efficiency of preeclampsia screening based on biomarkers of aneuploidy screening in first trimester
Yuan WANG ; Ya WANG ; Huirong TANG ; Honglei DUAN ; Ying ZHANG ; Jie LI ; Yan ZHANG ; Xiaodong YE ; Zhiyin WANG ; Guangfeng ZHAO ; Yali HU ; Mingming ZHENG
Chinese Journal of Perinatal Medicine 2022;25(8):601-608
Objective:To evaluate the performance of biomarkers in aneuploidy screening in the first trimester-pregnancy associated plasma protein A(PAPP-A) combined with Fetal Medicine Foundation (FMF)'s competing risk model in screening preeclampsia among our population.Methods:This study was based on a prospective cohort of singleton pregnant women who underwent aneuploidy screening in the first trimester in Nanjing Drum Tower Hospital from January 2017 to September 2020. Mean arterial pressure (MAP), uterine artery pulsatility index (UtA-PI), and PAPP-A were converted into multiples of median (MoM) using the algorithm disclosed on the website of the FMF (fetalmedicine.org). The predictive outcomes of maternal factors alone or in combination with MAP, UtA-PI, and PAPP-A (alone or in combination) were calculated. Chi-square test, Fisher's exact test or rank sum test were used for comparison among groups and Bonferroni method for pairwise comparisons. Receiver operating characteristic (ROC) curve was used to evaluate the screening efficiency and to calculate the sensitivities of predicting preeclampsia, term and preterm preeclampsia at false-positive rates of 5% and 10%. The predictive performance of this model was further compared to the screening strategy that was recommended in Diagnosis and treatment of hypertension and pre-eclampsia in pregnancy: a clinical practice guideline in China (2020). Results:Among the 5 144 singleton pregnancy women who were recruited in the cohort, 4 919 cases were included and analyzed in this study. A total of 223 cases were diagnosed as preeclampsia (4.5%), including 55 preterm (1.1%) and 168 term preeclampsia (3.4%). The median of MoM values of MAP, UtA-PI, and PAPP-A in the non-preeclampsia group were around 1.0±0.1. Statistical significance was observed in the difference of MAP, UtA-PI, and PAPP-A Mom between women with preterm preeclampsia and those without preeclampsia [1.061 (0.999-1.150) vs 0.985 (0.935-4.043), 1.115 (0.873-1.432) vs 1.039 (0.864-1.236), 0.820 (0.493-1.066) vs 1.078 (0.756-1.508)], which was also seen in the difference of MAP and PAPP-A Mom between women with term preeclampsia and those without preeclampsia [1.065 (1.002-1.133) vs 0.985 (0.935-4.043), 1.007 (0.624-1.393) vs 1.078 (0.756-1.508)] (all P<0.025). The combination screening with maternal factors+MAP+UtA-PI+PAPP-A was noted for the best efficiency. In predicting preeclampsia preterm and term preeclampsia at the false-positive rate of 10%, the sensitivity of the model was 53.0%, 76.4% and 44.6% respectively. Using the screening method recommended in Diagnosis and treatment of hypertension and pre-eclampsia in pregnancy: a clinical practice guideline in China(2020), the proportion of people at high risk of preeclampsia was 5.9% (290/4 919), and the sensitivity for predicting preterm preeclampsia was 25.5% (14/55), which was significantly lower than the combination screening with maternal factors+MAP+UtA-PI+PAPP-A [65.5% (36/55)] when using the same proportion of high-risk population. Conclusion:The preeclampsia screening model based on aneuploidy screening biomarkers in the first trimester--PAPP-A in combination with materral factors, MAP, UtA-PI, can effectively screen preterm preeclampsia in the local population without increasing the laboratory costs.
4.Clinical research of Qingfei-Xiaoyan Decoction combined with conventional western medicine in patients with acute exacerbation of chronic obstructive pulmonary disease
Cunming LI ; Zhenqin ZHENG ; Guangfeng ZHAO ; Ziwei LIN
International Journal of Traditional Chinese Medicine 2020;42(4):343-346
Objective:To observe the clinical efficacy of Qingfei-Xiaoyan Decoction combined with conventional western medicine in patients with acute exacerbation of chronic obstructive pulmonary disease (COPD). Methods:Eighty-two patients with acute exacerbation of COPD were selected in Bozhou People's Hospital from January 2017 to June 2018 and randomly divided into a control group and a treatment group (41 in each group) using randomized number table method. Patients in the control group were treated with conventional western medicine and those in the treatment group with Qingfei-Xiaoyan Decoction on the basis of the control group for 2 weeks. Dyspnea of patients was evaluated with the modified British Medical Research Council dyspnea questionnaire (mMRC). Impact of the disease was measured with the COPD Assessment Test (CAT). The serum levels of C-reactive protein (CRP) and procalcitonin (PCT) were measured by ELISA. Results:The total efficacy rate in trementat group 95.1% (39/41) was significantly higher than that in the control group 73.2% (30/41) ( χ2=4.999, P=0.025). After the treatment, the scores of mMRC (0.63 ± 0.07 vs. 0.95 ± 0.12; t=7.921, P<0.01) and CAT (9.18 ± 0.12 vs. 14.01 ± 1.56; t=11.359, P<0.01) in the treatment group were significantly lower than those in the control group. After the treatment, the forced expiratory volume in one second (FEV1) percentage predicted (51.05% ± 5.63% vs. 45.77% ± 5.31%; t=10.453, P<0.01) and FEV1/forced vital capacity (FVC) (59.15 ± 6.44 vs. 54.24 ± 6.02; t=5.621, P<0.01) in the treatment group were significantly higher than those in the control group. After the treatment, the serum levels of CRP (8.06 ± 0.87 mg/L vs. 10.55 ± 1.21 mg/L; t=10.216, P<0.01) and PCT (4.20 ± 0.48 μg/L vs. 6.33 ± 0.69 μg/L; t=7.004, P<0.01) in the treatment group were significantly lower than those in the control group. Conclusions:Qingfei-Xiaoyan Decoction can inhibite inflammation, improve symptoms and lung function, increase the efficacy in patients with acute exacerbation of COPD.
5.Estrogen Promotes cAMP Production in Mesenchymal Stem Cells by Regulating ADCY2
Guangfeng ZHAO ; Xiujun LI ; Huishuang MIAO ; Shiwen CHEN ; Yayi HOU
International Journal of Stem Cells 2020;13(1):55-64
Background and Objectives:
The maternal-fetal interface is an important source of mesenchymal stem cells (MSCs), and it is influenced by high levels of estradiol (E2) during pregnancy. It is highly important to study the role of E2 in MSCs for both clinical application and understanding of the mechanisms underlying pregnancy related diseases.
Methods:
and Results: In this study, differently expressed genes (DEGs) were found in the MSCs after exposure to E2. Then, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis of DEGs was performed and the integrated regulatory network of DEGs-miRNA was constructed. A total of 390 DEGs were found in the MSCs exposed to E2, including 164 upregulated DEGs (e.g. ADCY2, VEGFA and PPY) and 226 downregulated DEGs (e.g. KNG1, AGT and NPY). Additionally, 10 miRNAs (such as miR-148A/B, miR-152, miR-182) identified the integrated regulatory network of DEGs-miRNAs. Among them, the expression of ADCY2 was significantly upregulated, and this was associated with multiple changed genes. We confirmed that the expression of ADCY2 is significantly promoted by E2 and subsequently promoted the production of cAMP in MSCs. We also found that E2 promoted ADCY2 expression by inhibiting miR-152 and miR-148a.
Conclusions
E2 promotes the expression of cAMP through miR-148a/152-ADCY2 in MSCs. It is suggested that E2 plays a key role in the growth and function of MSCs.
6.Induced Intermediate Mesoderm Combined with Decellularized Kidney Scaffolds for Functional Engineering Kidney
Jianye ZHANG ; Kailin LI ; Feng KONG ; Chao SUN ; Denglu ZHANG ; Xin YU ; Xuesheng WANG ; Xian LI ; Tongyan LIU ; Guangfeng SHAO ; Yong GUAN ; Shengtian ZHAO
Tissue Engineering and Regenerative Medicine 2019;16(5):501-512
BACKGROUND: Chronic kidney disease is a severe threat to human health with no ideal treatment strategy. Mature mammalian kidneys have a fixed number of nephrons, and regeneration is difficult once they are damaged. For this reason, developing an efficient approach to achieve kidney regeneration is necessary. The technology of the combination of decellularized kidney scaffolds with stem cells has emerged as a new strategy; however, in previous studies, the differentiation of stem cells in decellularized scaffolds was insufficient for functional kidney regeneration, and many problems remain. METHODS: We used 0.5% sodium dodecyl sulfate (SDS) to produce rat kidney decellularized scaffolds, and induce adipose-derived stem cells (ADSCs) into intermediate mesoderm by adding Wnt agonist CHIR99021 and FGF9 in vitro. The characteristics of decellularized scaffolds and intermediate mesoderm induced from adipose–derived stem cells were identified. The scaffolds were recellularized with ADSCs and intermediate mesoderm cells through the renal artery and ureter. After cocultured for 10 days, cells adhesion and differentiation was evaluated. RESULTS: Intermediate mesoderm cells were successfully induced from ADSCs and identified by immunofluorescence and Western blotting assays (OSR1 + , PAX2 +). Immunofluorescence showed that intermediate mesoderm cells differentiated into tubular-like (E-CAD + , GATA3 +) and podocyte-like (WT1 +) cells with higher differentiation efficiency than ADSCs in the decellularized scaffolds. Comparatively, this phenomenon was not observed in induced intermediate mesoderm cells cultured in vitro. CONCLUSION: In this study, we demonstrated that intermediate mesoderm cells could be induced from ADSCs and that they could differentiate well after cocultured with decellularized scaffolds.
Animals
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Blotting, Western
;
Fluorescent Antibody Technique
;
Humans
;
In Vitro Techniques
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Kidney
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Mesoderm
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Nephrons
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Rats
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Regeneration
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Renal Artery
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Renal Insufficiency, Chronic
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Sodium Dodecyl Sulfate
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Stem Cells
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Ureter
7.Effect of surfactant on respiratory distress syndrome: evaluation by bedside pulmonary ultrasonography
Fang YOU ; Xueyun REN ; Fenghai NIU ; Jing ZHAO ; Guangfeng QIANG ; Meiqin XIANG
Chinese Journal of Neonatology 2019;34(1):34-37
Objective To study the relationship between the lung ultrasonography and the chest X-ray and to study the value of lung ultrasonography score (LUS) in evaluating the effect of pulmonary surfactant (PS) on respiratory distress syndrome (RDS) of newborn.Method Preterm infants admitted to the neonatal intensive care unit of our Hospital from January 2016 to December 2017 and diagnosed with RDS were prospectively studied.LUS examinations were performed prior to,and within the first 6~12 hours after surfactant administration,chest X-rays were also performed at the same time so as to evaluate the effects of surfactant replacement therapy and the correlation between the lung ultrasonography and the chest X-rays.Lung ultrasonography findings at a total of six sites,with three sites in each lung were scored based on the presence of normal finding,the amount of B-lines and subpleural consolidations.Result A total of 45 preterm infants with RDS were enrolled.The cases of X-ray grades Ⅰ,Ⅱ,Ⅲ and Ⅳ before PS administration were 5 cases,21 cases,12 cases and 7 cases respectively.The scores of LUS 0~6,7~12,13~ 18 were 5 cases,37 cases and 3 cases respectively,and the median of LUS was 10 points.Chest X-ray grades Ⅰ,Ⅱ,Ⅲ and Ⅳ within 6~12 hours after PS administration were 18 cases,17 cases,8 cases and 2 cases respectively.LUS of 0~6,7~12,13~18 were 21 cases,20 cases and 4 cases respectively.The median of LUS after PS was 7 points.LUS after PS application was significantly lower than that before PS application (P<0.001).The LUS was positively correlated with the grades of X-ray before and after surfactant administration (before surfactant administration r =0.688,P<0.001,after surfactant administration r =0.777,P<0.001).Conclusion LUS is positively correlated with the grade of chest X-ray and might enable an early detection of the surfactant replacement therapy effects in RDS.Further studies are necessary to define the role of LUS in this field.
8.Pneumothorax in critically ill neonates: diagnosis using bedside ultrasonography
Xueyun REN ; Jing ZHAO ; Fenghai NIU ; Fang YOU ; Guangfeng QIANG ; Na LIU
Chinese Journal of Neonatology 2019;34(6):439-442
Objective To study the value of lung ultrasonography (LUS) in the diagnosis of pneumothorax in critically ill neonates.Method The neonates admitted to our NICU and suspected to have pneumothorax were prospectively enrolled from June 2017 to December 2018.All eligible infants received both LUS examination and chest X-ray.The characteristics of LUS imaging was analyzed based on the chest X-ray which was used as the golden standard for the diagnosis of pneumothorax.The sensitivity,specificity,positive predictive value and negative predictive value of LUS is computed.The duration of LUS and chest X-ray were compared.The outcome and complications were also observed.Result Fifty neonates with suspected pneumothorax were collected.Among them,pneumothorax was confirmed with chest X-ray in 31 neonates (62.0%).Ultrasound signs of pneumothorax included absence of lung sliding (100%),absence of B lines (100%),stratosphere sign (100%) were observed in all of the 31 neonates.Presence of lung point was also observed in 90.3% of the patients.The sensitivity,specificity,positive predictive value,negative predictive value and X-ray coincidence rate of LUS in the diagnosis of pneumothorax were 100%.LUS and chest X-ray examination took (5.6 ±5.1) min and (20.1 ± 12.2) min,respectively,the difference was statistically significant (P < 0.05).All 31 infants with pneumothorax survived.15 infants underwent closed thoracic drainage after emergency thoracic puncture or aspiration assisted by LUS.No postoperative complications occurred.Conclusion LUS showed high accuracy,sensitivity and specificity in detecting pneumothorax in critically ill neonates.It is simple to operate and can guide clinical rescue more promptly and quickly.
9. The correlation between lung ultrasound score and neonatal critical illness score and its clinical predictive value
Guangfeng QIANG ; Jing ZHAO ; Lanlan MENG ; Fenghai NIU ; Xueyun REN
Chinese Journal of Ultrasonography 2019;28(9):748-752
Objective:
To explore whether the lung ultrasound(LUS) score can be used to assess and predict the criticality of neonates with pulmonary disease at an early stage.
Methods:
The newborns born in the obstetrics department of Affiliated Hospital of Jining Medical University from April to October 2018 were transferred to the neonatal intensive care unit due to respiratory distress. The children underwent LUS examination and scoring at 2 hours after birth. The correlation analysis were performed between LUS score and neonatal critical illness score (NCIS ), NCIS+ single index, respectively. And the ROC curve was used to analyze the value of LUS score in predicting neonatal criticality.
Results:
①The LUS score of non-critical neonates was significantly lower than that of critically ill newborns, the difference was statistically significant (
10.The correlation between lung ultrasound score and neonatal critical illness score and its clinical predictive value
Guangfeng QIANG ; Jing ZHAO ; Lanlan MENG ; Fenghai NIU ; Xueyun REN
Chinese Journal of Ultrasonography 2019;28(9):748-752
Objective To explore whether the lung ultrasound( LUS) score can be used to assess and predict the criticality of neonates with pulmonary disease at an early stage . Methods T he new borns born in the obstetrics department of Affiliated Hospital of Jining M edical University from April to October 2018 were transferred to the neonatal intensive care unit due to respiratory distress . T he children underwent LUS examination and scoring at 2 hours after birth . T he correlation analysis were performed between LUS score and neonatal critical illness score ( NCIS ) ,NCIS +single index ,respectively . And the ROC curve was used to analyze the value of LUS score in predicting neonatal criticality . Results ①T he LUS score of non‐critical neonates was significantly lower than that of critically ill newborns , the difference was statistically significant ( P =0 .005) ; LUS score was an independent risk factor for critical neonates ( OR=1 .71 ,95%CI :1 .059-2 .765 , P = 0 .028 ) . ② T he correlation coefficient between LUS score and NCIS was -0 .48 ( P =0 .002) . T he correlation coefficient between the LUS score and the NCIS + single index was -0 .44 ( P=0 .005) . ③T he area under the ROC curve of LUS score predicting neonatal criticality was 0 .88 ( 95%CI :0 .725-0 .965 , P <0 .000 1) ,the optimal diagnostic threshold was 6 points with sensitivity of 80% and specificity of 100% . Conclusions The LUS score at a postnatal age of 2 hours after birth can early assess and predict the criticality of neonates with pulmonary disease . And the LUS score greater than 6 has the highest diagnostic value .

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