1.Induction of preeclamptic rat model by injection of ultra-low-dose lipopolysaccharide in early pregnancy
Jinmei WANG ; Mingming ZHENG ; Jianjun ZHOU ; Zhenyu DIAO ; Yali HU
Chinese Journal of Perinatal Medicine 2012;(11):689-695
Objective To establish an animal model of preeclampsia by injecting ultra-low-dose lipopolysaccharide (LPS) to rats in early pregnancy,and to lay the foundation for further study on mechanisms of preeclampsia.Methods Twenty-four pregnant rats were divided into six groups according to the random number table and were injected with LPS 0.3,0.5,0.7,1.0,2.0 μg/kg or saline 2 ml respectively through tail veins on day 5 of pregnancy.The differences in blood pressure,urinary protein and pathological changes in placenta among groups were compared to confirm the suitable dose of LPS for establishing preeclamptic model.Then another 19 pregnant rats were injected with the chosen dose of LPS slowly through tail veins on day 5 of pregnancy; 15 of which were chosen as model group; the other four were chosen as postpartum group.Three non-pregnant rats were as non-pregnant group.Besides,another 15 pregnant rats were injected with saline as pregnant control group.Systolic blood pressure,urinary protein excretion,placental weight,fetal weight,serum white blood cell counts,blood platelet counts,plasma anti-thrombin-Ⅲ content,D-dimer content were examined and compared among groups with one way analysis of variance; histopathologic studies were also done on the placentas,kidneys and aortas of the rats.Results (1) Placental weight of LPS 0.3 μg/kg group increased compared with control group.One pregnant rats(1/4) in LPS 1.0 μg/kg group and LPS 2.0 μg/kg group died on day 16 of pregnancy as a result of vaginal bleeding.Systolic blood pressure of LPS 0.5 μg/kg group rose steadily,while no significant changes were found in other groups.Urinary protein increased in all LPS groups,while urinary protein of LPS 0.7 μg/kg group and LPS 1.0 μg/kg group peaked on day 12 of pregnancy and then decreased; urinary protein of LPS 0.5 μg/kg group increased most significantly,and fetus in LPS 0.5,0.7 and 2.0 μg/kg groups had lighter body weight.So LPS 0.5 μg/kg was chosen as the suitable dose to establish preeclamptic model.(2)Compared with pregnant control group,model group had higher systolic blood pressure [(124.89±1.79) mm Hg vs (119.02±1.80) mm Hg,LSD test,P=0.03] from day 6 of pregnancy,more urinary protein [(2.02±0.29) mg vs (1.11±0.18) mg,LSD test,P=0.00] from day 9 of pregnancy,more absorbed embryos [3.6% (7/194) vs 0.0% (0/200),Fisher exact test,P=0.01] at day 20 of pregnancy,higher incidence of placenta bleeding [4.1% (8/194) vs 0.0% (0/200),Fisher exact test,P=0.00] and fetal growth restriction [13.9% (27/194) vs 6.0% (12/200),X2=6.92,Fisher exacttest,P=0.01].Model group showed more inflammatory cells infiltration in the placenta,more glomerular mesangial cells,swelling and desquamated of renal tubular epithelial cells compared to control group.Blood pressure and urinary protein of the model group recovered to the baseline at the sixth day of postpartum,and no changes in blood pressure and urinary protein were found in non-pregnant rats.Conclusions Injection of LPS 0.5 μg/kg on day 5 of pregnancy through tail veins could induce the clinical symptoms of preeclampsia in rats,which might be an ideal model for further preeclampsia research.
2.Relationship between proliferation of prostate cancer cells and androgen receptor expression
Xinwei DIAO ; Mingfu YE ; Zaigen ZHANG ; Yali WANG
Journal of Third Military Medical University 2003;0(17):-
Objective To explore the relationship between proliferation activity-related factors and androgen receptor (AR) expression in 113 cases of human prostate cancer (PCa). Methods AR and the proliferation activity-related factors (PCNA, Ki67, EGFR) in PCa were detected by immunohistochemistry and light/(electron) microscopy. Results Cell proliferation in AR-negative groups of PCa was significantly stronger than in AR-positive groups. Conclusion AR aberrant expression may promote proliferative ability of PCa, cell growth and heterogeneity significantly, which may be one of reasons that PCa is difficult to be cured.
3.Different expression of microRNA-155 and cysteine-rich 61 in human placentas from severe preeclamptic and normal pregnancies
Zhiqun WANG ; Li SU ; Zhenyu DIAO ; Jianjun ZHOU ; Yimin DAI ; Yanqing ZHANG ; Yali HU
Chinese Journal of Perinatal Medicine 2010;13(5):403-407
Objective To investigate the different expression of microRNA-155 (miR-155) and cysteine-rich 61 (CYR61) in human placentas between severe preeclamptic and normal pregnancies.Methods Placentas were obtained from severe preeclamptic and healthy control pregnant women (n=18 for each group) at 36~40 gestational weeks. The expressions of miR-155 and CYR61 mRNA were assessed by real-time quantitative reverse transcription-polymerase chain reaction, and the levels of CYR61 protein were tested by Western blot. Results Compared with the control group, the miR-155 expression was increased in placentas from severe preeclampsia groups ( 165. 7 ± 16. 4 vs 527.9±49.1,t=7.00, P<0.01), and the CYR61 mRNA expression (31.7±2.7 vs 16.4±1.2,t=5.10,P<0. 01), as well as the CYR61 protein expression (36.4±1.5 vs 19.7±1.2,t=36.26, P<0.01 ) were decreased. There was a significantly negative correlation between the expression of miR-155 and CYR61 mRNA within both groups (preeclamptic group: r=-0.52, P<0.05;control:r=-0.57, P<0.05). Conclusions Up-regulation of placental miR-155 in severe preeclampsia may be related to the decreased expression of CYR61. Both miR-155 and CYR61 may contribute to the disorders of placental angiogenesis in severe preeclampsia in human.
4.Clinical significance of expressions of vascular endothelial growth factor and matrix metalloproteinase-9 in prostate carcinoma
Qinwen WANG ; Xinwei DIAO ; Zhengtang CHEN ; Mingfu YE ; Jianping XU ; Yali WANG
Journal of Third Military Medical University 2003;0(19):-
Objective To explore the relationship between the expressions of vascular endothelial growth factor(VEGF) and matrix metalloproteinase-9(MMP-9) in prostate cancer.Methods The expression of VEGF and MMP-9 in 148 samples of pathologically verified human prostate cancer(PCa) and 10 samples of benign prostatic hyperplasia(BPH) were detected by immunohistochemical staining.Results VEGF was overexpressed in 98 of 148 tumor tissues(66.2%) and MMP-9 was overexpressed in 58 of 148 tumor tissues(39.2%).Both of them were significantly higher than those in BPH(P0.05).But the expression of VEGF was correlated with that of MMP-9.Conclusion The expressions of VEGF and MMP-9 are closely associated with the development of prostate cancer.Therefore VEGF and MMP-9 may be clinically useful as predictor of patients'survival.
5.Isolation and identification of placental exosomes from maternal serum
Yujing LI ; Zhenyu DIAO ; Pingping XUE ; Li SHEN ; Ping GONG ; Guijun YAN ; Yali HU
Journal of Medical Postgraduates 2015;(6):632-636
Objective During pregnancy , exosomes can be released from the placenta into maternal circulation and play im-portant roles in normal pregnancy or placenta-related diseases .We aimed to establish a simple and efficient method for isolating and i-dentifying placental exosomes from maternal serum and lay a foundation for the studies of pregnancy -related diseases . Methods Using sucrose gradient centrifugation with 8% PEG6000 precipitation twice , we isolated and purified placenta-derived exosomes from normal maternal serum and detected their molecular markers CD 63 , CD81 and PLAP by Western blot , followed by silver staining anal-ysis of the protein profile of the exosome pellet .We identified the morphology of the placenta-derived exosomes by transmission electron microscopy ( TEM) and measured the size and distribution of the particles by dynamic light scattering ( DLS) . Results Silver stai-ning of the protein profiles of the exosomes after sucrose gradient centrifugation clearly revealed the bands of the protein molecules . Western blot showed the expressions of CD 63, CD81, and PLAP in the 21-34%density layer, which demonstrated the presence of serum placental exosomes mainly in the 1.09-1.16 g/mL density layer.TEM exhibited that the placenta-derived exosomes were round or oval cup-shaped, specifically expressing PLAP, and the particles were uniform in size, with a mean diameter of (41.79 ±11.94) nm. Conclusion A simple, fast, and efficient method was successfully established for isolating placenta-derived exosomes from ma-ternal serum, which provides a basis for studying the roles of placental exosomes in normal pregnancy and placenta -related diseases.
6.Pravastatin inhibits microRNA-155 expression and improves functions of lipopolysaccharide-treated human extravillous trophoblast cells
Zhiyin WANG ; Muyi YANG ; Xiaoyu DUAN ; Zhenyu DIAO ; Hailin DING ; Yanfang PENG ; Yi LEI ; Guangfeng ZHAO ; Dan LIU ; Yali HU
Chinese Journal of Perinatal Medicine 2017;20(10):705-711
Objective To investigate the effects ofpravastatin on the expression ofmicroRNA-155 (miR-155) and the functions of lipopolysaccharide (LPS)-treated extravillous trophoblast cells.Methods In vitro cultured HTR-8/SVneo cells were divided into the following groups:control group,enhanced plasmid with green fluoscent protein (pEGFP)-miR-155 group (transfected with green fluorescent protein-tagged miR-155),LPS group (treated with 100 ng/mL of LPS),miR-155 inhibitor+LPS group,pravastatin+LPS group (treated with 100 ng/mL of LPS following pretreatment with 12.50,25.00,50.00 and 100.00 μ g/ml of pravastatin),and pravastatin+pEGFP-miR-155 group (transfected with pEGFP-miR-155 following pretreatment with 50 μ g/ml of pravastatin).Levels of miR-155 in HTR-8/SVneo cells treated with different strategies were measured by real-time polymerase chain reaction.Expression of phosphorylated JunB (p-JunB) and p-FosB proteins was analyzed by Western blotting.Migration,invasion and apoptosis of HTR-8/SVneo cells were also analyzed.All data were analyzed with t test.Results (1) Compared with the control group,HTR-8/SVneo cells in the pEGFP-miR-155 group were characterized by shorter migration distance [(274.70± 18.82) vs (181.00±8.62) μ m],less transmembrane cells [(123.00±4.36) vs (63.00±6.08)] and enhanced apoptosis [(5.40± 0.68)% vs (9.27±0.68)%] (all P<0.05).(2) Compared with the LPS group,the miR-155 inhibitor+LPS group showed longer migration distance of HTR-8/SVneo cells [(166.30±5.07) vs (242.00±18.07) μm,P<0.05],more transmembrane cells [(71.67±6.12) vs (109.00±7.81),P<0.05] and decreased cell apoptosis [(14.40±1.69)% vs (6.23± 0.44)%,P<0.05].(3) Expression of miR-155 at mRNA level in the LPS group was increased as compared with that of the control group (1.65 0.07 vs 0.79±0.12,P<0.05).Compared with the LPS group,pretreatment with 12.50,25.00,50.00 and 100.00 μ g/ml of pravastatin decreased the expression of miR-155 at mRNA level [(1.14±0.10),(1.02±0.10),(0.74±0.15) and (1.140.02)],especially at the concentration of 50 μμ g/ml (all P<0.05).(4) Expression ofp-JunB and p-FosB proteins in the control,LPS and pravastatin (50 μ g/ml)+LPS groups were (0.33 ±0.06) vs (0.37±0.07),(1.22±0.20) vs (0.80±-0.13),and (0.31 ±0.02) vs (0.21 ±0.05),respectively,showing higher expression level in both p-JunB and p-FosB proteins in the LPS group compared with that of the other two groups (all P<0.05).(5) Compared with the LPS group,the pravastatin (50 μμ g/ml)+LPS group showed increased migration distance [(166.30±5.07) vs (246.80± 13.42) μ m,P<0.05],increased numbers of transmembrane cells [(71.67 ± 6.12) vs (95.33 ± 2.73),P<0.05] and decreased cell apoptosis [(14.40± 1.69) vs (6.05 ± 0.35)%,P<0.05].(6) Compared with the pEGFP-miR-155 group,the pravastatin (50.00.00 μμ g/mL)+pEGFP-miR-155 group showed longer migration distance [(197.50± 13.86) vs (275.80± 13.63) μ m,P<0.05],more transmembrane cells [(52.67±5.49) vs (125.00±6.66),P<0.05] and lower rate of cell apoptosis [(8.90± 1.00) vs (5.05±0.35)%,P<0.05].Conclusions Pretreatment of extravillous trophoblast cells with pravastatin can protect them from apoptosis and loss of migratory and invasive abilities through inhibiting the activation of AP-1 and down-regulating the expression of miR-155,which may be a mechanism that inhibits the development of preeclampsia.
7.Triptolide inhibits activity of human cervical microvascular endothelial cells
ZHANG Yali ; DIAO Yunyun ; ZHANG Chunze
Chinese Journal of Cancer Biotherapy 2019;26(2):196-199
Objective: To investigate the inhibitory effect of Triptolide on vasculogenesis of human cervical microvascular endothelial cells, and to explore the mechanism. Methods: Human cervical microvascular endothelial cells (HCerMECs) were used as research subject, and treated with different concentrations of Triptolide (0, 5, 10, 20 and 40 ng/ml) i n v i t r o . The effect of Triptolide on cell proliferation was determined by CCK-8, the cell migration ability was detected by Transwell assay while the expression of vascular endothelial growth factor (VEGF) was examined by western blotting. Results: Triptolide inhibited the proliferation and migration of HCerMECs in a dose-dependent manner ( P <0.05). In addition, Triptolide could inhibit the expression of VEGF in HCerMECs in a concentration-dependent manner. Conclusions: Triptolide could inhibit the proliferation and migration activity of HCerMECs which is related with the suppression of VEGF expression.
8.Application of Single-hole Thoracoscopic Surgery Combined with ERAS Concept for Respiratory Function Exercise in Perioperative Period of Lung Cancer.
Qingtong SHI ; Yali DIAO ; Jun QIAN
Chinese Journal of Lung Cancer 2020;23(8):667-672
BACKGROUND:
Lung cancer is currently the leading malignant tumor in both domestic and foreign morbidity and mortality. Surgical treatment is the main treatment option for lung cancer. The aim of this study is to explore the effects of enhanced recovery after surgery (ERAS) combined with respiratory function exercise combined with single-hole thoracoscopic surgery on lung cancer patients with postoperative pulmonary complications, postoperative pain, time to get out of bed, time to extubation and length of hospital stay.
METHODS:
A total of 240 patients who underwent endoscopic lung cancer surgery at the Affiliated Hospital of Yangzhou University and the Yancheng First People's Hospital from October 2017 to October 2019 were randomly divided into 4 groups, with 60 patients in each group. Patients in group A underwent single-hole thoracoscopic surgery, and preoperatively performed ERAS concept education and respiratory function training; group B used conventional 3-hole thoracoscopic surgery, and performed ERAS concept education and respiratory function training before operation; group C used conventional 3-hole thoracoscopic operation surgery, routine hospitalization education and nursing guidance, routine respiratory function training, no preoperative ERAS concept education; group D used single-hole thoracoscopic surgery, routine hospitalization education and nursing guidance, routine respiratory function training, no preoperative ERAS concept mission. The number of postoperative pulmonary complications, postoperative pain, time to get out of bed, extubation time, and hospital stay were recorded in the four groups.
RESULTS:
Compared with the groups B, C, and D, the incidence of pulmonary complications was significantly reduced, and the time to get out of bed, extubation time, and hospital stay were significantly shortened in group A. Compared with groups B, C, the postoperative pain was significantly reduced in group A. Compared with group C, the pulmonary complications were significantly reduced, and the time to get out of bed, extubation time and hospital stay were significantly shortened in group B. The differences were statistically significant (P<0.05). There was no significant difference in postoperative pain between group A and group D, group B and group C (P>0.05).
CONCLUSIONS
For patients with single-hole thoracoscopic lung cancer surgery, the ERAS concept guidance can effectively reduce the incidence of pulmonary complications and postoperative pain, shorten the time to get out of bed, the time to extubate, and the length of hospital stay.
9.Clinical Value of Four-hole Unilateral Dissecting Lobectomy and Mediastinal Lymph Node Dissection in the Treatment of Early Non-small Cell Lung Cancer.
Hong GUO ; Yali DIAO ; Huangxin FAN ; Qingquan LUO
Chinese Journal of Lung Cancer 2018;21(8):578-582
BACKGROUND:
Thoracoscopic lobectomy combined with mediastinal lymph node dissection has been considered as one of the standard surgical procedures for early lung cancer. After 20 years of development, thoracoscopic lobectomy has reached a consensus on reliability and minimally invasive. At present, thoracoscopic lobectomy has a variety of incisions, which gradually evolve into four holes based on three holes, and two or one hole as the operative approach. The aim of this study was to evaluate the clinical value of four-hole unilateral dissecting lobectomy and mediastinal lymph node dissection in the treatment of non-small cell lung cancer (NSCLC). The aim of this study was to investigate the clinical value of anatomical lobectomy with mediastinal lymphadenectomy under four-hole completely video-assisted thoracoscopic surgery (C-VATS) in the treatment of non-small cell lung cancer.
METHODS:
The patients undergoing lobectomy with mediastinal lymphadenectomy for NSCLC were identified in the Department of Thoracic Surgery, Yangzhou First People's Hospital, Yangzhou University from March 2015 to July 2016. Preoperative clinical diagnosis of peripheral-type early NSCLC. The patients were randomly divided into four-hole monophasic group (experimental group) and three-hole group (control group) according to the number of hospitalization before surgery. According to inclusion and exclusion criteria, the 39 cases assign in experimental group and 34 cases in the control group, including 36 males and 37 females; aged 38 to 84 years. The mean operation time, average blood loss, lymph node dissection group, average drainage, average extubation time and postoperative complications were compared between the two groups for statistical analysis.
RESULTS:
The two groups of patients were successfully completed surgery, no death after surgery. Mean bleeding in the two groups, the number of lymph node dissection group, the average postoperative drainage, the average time of extubation, postoperative complications, with no significant difference. The average operation time of the four-hole unidirectional group was shorter than that of the three-hole group. The difference was statistically significant (P<0.05).
CONCLUSIONS
The safety and efficacy of a four-hole one-way operation under VATS are satisfactory. The operation is smooth during operation, which shortens the course of operation and deserves the clinical promotion.
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Adult
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Aged
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Aged, 80 and over
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Carcinoma, Non-Small-Cell Lung
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surgery
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Female
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Humans
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Lung Neoplasms
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surgery
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Lymph Node Excision
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methods
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Male
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Mediastinum
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Middle Aged
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Operative Time
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Pneumonectomy
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methods
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Retrospective Studies
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Thoracic Surgery, Video-Assisted
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Time Factors