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OBJECTIVE: To investigate the correlation between coagulation parameters and pelvic adhesions in endometriosis.METHODS: The clinical data of 408 patients with endometriosis who underwent surgery in Women's Hospital School of Medicine Zhejiang University from June 2016 to February 2018 were retrospectively analyzed to evaluate the correlation between coagulation parameters and the severity of pelvic adhesions.RESULTS: In endometriosis cases,the levels of plasma fibrinogen(FIB)in patients with pelvic adhesions were significantly shorter than those without pelvic adhesions[median:3.1 g/L(range:2.7-4.5 g/L)vs. 2.8 g/L(range:2.6-3.2 g/L),respectively;P0.05). FIB was positively correlated with pelvic adhesion scores(r=0.248,P<0.01).In the judgment of pelvic adhesion,the area under the curve(AUC)of FIB was 0.630,and the cut-off value was 2.87 g/L.The incidence of adhesions in patients with FIB greater than 2.87 g/L increased(χ2=15.4,P<0.01).CONCLUSION: Coagulation parameters was correlated with the pelvic adhesions in endometriosis,especially fibrinogen,which may help to judge the severity of pelvic adhesions caused by endometriosis.
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Objective: To investigate the effect of cartilage tissue engineering scaffold PVA/ι-CA on the biological behavior of the ATDC-5 cells,and to evaluate its feasibility on constructing tissue engineering cartilage. Methods:The polyvinyl alcohol (PVA)and carrageenan were used to make the composite scaffold material PVA/ι-CA according to a certain proportion by physical blending technology and repeated freezing thawing method,and the porosity and pore size of PVA/ι-CA were detected.The ATDC-5 cells were seeded into the composite scaffold and its growth was observed; the expressions of collagen type Ⅱ in the ATDC-5 cells were tested by immunohistochemical staining and immunofluorescence staining; the morphology of the ATDC-5 cells was confirmed by Toluidine blue staining.The growth and secretion of extracellular matrix of the ATDC-5 cells were observed under scanning electron microscope (SEM);the proliferative rates of ATDC-5 cells in composite scaffold materials in negative control group (added with DMEM culture media)and experimental group (added with DMEM contain scaffold)were determined by MTT assay.The composite scaffolds were implanted subcutaneously in the SD rats.The histocompatibility and vascularization in vivo of the composite scaffolds were evaluated.Results:The average porosity of cartilage tissue engineering scaffold PVA/ι-CA was (86.88±3.88)%,and the average pore size was 20-40 μm.The HE staining results showed that the ATDC-5 cells grew well with the polygon and plumpness morphology. All the samples were stained positive for collagen type Ⅱ by immunohistochemistry and immunofluorescence staining,which verified the normal phenotype of chondrocytes on the scaffolds. All the sample were stained positive for toluidine blue staining,which verified ECM deposition of the ATDC-5 cells on the scaffolds.The number of the positive cells was significantly increased with the prolongation of time.After cultured for 7 d,few of the ATDC-5 cells presented polygonal;after cultured for 14 d,the ATDC-5 cells distributed more densely,and contacted with each other on the scaffold;after cultured for 21 - 28 d,the ATDC-5 cells filled the interconnected pores of the scaffolds,synthesizing a significant amount of neo-formed ECM.The proliferation of ATDC-5 cells in PVA/ι-CA grew fast during 7-14 d,and it became slow during 21-28 d;the difference was not statistically significant compared with control group (P >0.05).The subcutaneous implantation results showed the inflammatory reactions were slight at the early stage and eviated gradually,there was an increasing angiogenesis at the late stage,and the degradation and absorption of the meterial were slight.Conclusion:PVA/ι-CA composite material will be an ideal material for the cartilage tissue engineering.
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Objective To investigate the roles and mechanisms of siRNA targeted lipocalin 2 (Lcn 2) gene silencing on hypoxia-induced cell apoptosis in retinal ganglion cells (RGC-5).Methods RGC-5 cells were subjected to hypoxic condition for various time duration (0 h,4 h,8 h,12 h,24 h,48 h),and quantitative real-time PCR and Western blot were used to evaluate the expression of Lcn 2 in mRNA and protein levels.Cells were divided into 4 groups:control group,hypoxia group,siNC + hypoxia group,in which cells were transfected with negative control for siRNA,and then subjected to hypoxic condition for 24 h,and siLcn 2 + hypoxia group,in which cells were transfected with Lcn 2 siRNA,and then subjected to hypoxic condition for 24 h).Next,cell apoptotic rate and Caspase-3 activity were detected using ELISA.The fluorescence intensity of reactive oxygen species (ROS) was assayed using DCFH-DA kit,and mitochondrial membrane potential assay kit was used to evaluate the mitochondrial membrane potential.Finally,the expression levels of cleaved-Caspase-3 (c-Caspase-3),cleaved-PARP (c-PARP),Bax,Bcl-2 and cytochrome C (Cyto C) were detected using Western blot.Results In the hypoxia group,Lcn 2 mRNA level at 4 h,8 h,12 h,24 h and 48 h was higher than that at 0 h (all P ＜0.05).Meanwhile,Lcn 2 protein level at 12 h,24 h and 48 h was also higher than that at 0 h (all P ＜ 0.05).Cell apoptotic rate in the hypoxia group (138.33％ ± 13.76％) was significantly higher than that in the control group (99.66％ ± 2.86％) (P ＜ 0.05),and siLcn 2 + hypoxia group (105.02％ ± 8.60％) was lower than siNC + hypoxia group (142.33％ ± 6.54％) (P ＜ 0.05).In addition,compared with the control group,the relative activity of Caspase-3 and the relative expression of c-Caspase-3 and c-PARP in the hypoxia group were all upregulated (all P ＜ 0.05);whereas the relative activity of Caspase-3 and the relative expression of c-Caspase-3 and c-PARP in the siLcn2 + hypoxia group were downregulated compared with the siNC + hypoxia group (all P ＜ 0.05).Moreover,the fluorescence intensity of ROS in the hypoxia group (4.26 0.63) was more enhanced than that in the control group (1.03 ± 0.11) (P ＜ 0.05),and the siLcn2 + hypoxia group (3.10 ± 0.24) was lower than siNC + hypoxia group (4.73 ± 0.26) (P ＜ 0.05).Furthermore,mitochondrial membrane potential,Cyto C expression and the relative ratio of Bax to Bcl-2 in the siLcn2 + hypoxia group was lower than those in the siNC + hypoxia group.Conclusion Lcn 2 Lcn2 gene silencing can inhibit cell apoptosis and ROS production induced by hypoxia,which may be achieved by suppressing mitochondrial apoptosis signaling pathway.