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Objective To explore whether nano-vesicles derived from M1 macrophages (M1-NVs) can reprogram M2 macrophages into M1 phenotype and further affect the development of endometriosis (EMS). Methods Extracellular vesicles (EVs) were isolated from macrophage culture supernatant by differential centrifugation. Immunofluorescence cytochemistry was used to detect the expression of vimentin, CD31 and F4/80 to identify endometrial stromal cells (EMS-ESCs), HUVECs and polarized peritoneal macrophages of EMS patients. M1-NVs were prepared by filtering cell suspension through (5, 1, 0.4, 0.22)μm polycarbonate membrane filters after syringe aspiration at 0-4 DegreesCelsius. Flow cytometry was used to analyze the polarization of RAW264.7 mouse peritoneal macrophages in vitro, and reverse transcription PCR (RT-qPCR) was employed to detect mRNA expression of VEGF, CD86, interleukin-6 (IL-6), IL-1β, tumor necrosis factor α (TNF-α), arginase 1 (Arg1), CD163, CD206, and IL-10. PKH67-labeled M1-NVs were co-cultured with EMS-ESCs, HUVECs and macrophages. And tubule formation experiments were conducted to assess the impact of M1-NVs on the tubule formation of HUVECs. TranswellTM invasion and migration assays were employed to evaluate changes in the migration and invasion abilities of EMS-ESCs. Results By monitoring the contents of NVs, it was found that NVs contained much more protein and other bioactive particles than the same amount of EVs; immunofluorescence staining results showed that PKH67 labeled M1-NVs were internalized by EMS-ESCs, HUVECs and macrophages when co-cultured. The results of flow cytometry and RT-qPCR multi-target analysis showed that after treatment with different concentrations of M1-NVs or M0-NVs, 20 μg/mL of M1-NVs could effectively reprogram M2 macrophages into M1 macrophages compared with M0-NVs. TransewellTM results showed that compared with the blank group and M0-NVs group, the number of EMS-ESCs migrating from the upper chamber to the lower chamber after M1-NV treatment was significantly reduced, while the number of EMS-ESCs treated with M2NVs increased significantly. The invasion situation was similar to the migration situation, indicating that M1-NVs directly or indirectly inhibited invasion, migration and tubule formation of EMS-ESCs in vitro. Conclusion M1-NVs reprogrammes M2 macrophages into M1 macrophages by internalization of primary cells and macrophages, thereby inhibiting invasion, migration and angiogenesis of EMS-ESCs, and further hindering the occurrence and development of EMS.
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Female , Humans , Animals , Mice , Endometriosis , Macrophages , Macrophages, Peritoneal , Coculture TechniquesABSTRACT
Objective To investigate the expression of long non-coding RNA ubiquitin-specific peptidase 30 antisense RNA 1 (lncRNA USP30-AS1) and its relationship with immune infiltration in ovarian serous cystadenocarcinoma (OSC), and to determine its prognostic role in OSC. Methods The Cancer Genome Atlas (TCGA) database was utilized to retrieve the expression of USP30-AS1 and clinical information of 384 OSC patients. Wilcoxon rank-sum test was employed to compare the expression of USP30-AS1 between OSC and normal ovarian tissues. Logistic regression analysis was conducted to assess the relationship between clinical pathological features and USP30-AS1. Gene set enrichment analysis (GSEA) and single-sample gene set enrichment analysis (ssGSEA) were performed to investigate enrichment pathways and functions and quantify the degree of immune cell infiltration in USP30-AS1. Based on the expression level of long non-coding RNA (lncRNA) USP30-AS1, the samples were divided into high and low expression groups according to the expression mean. Log-rank tests, univariate and multivariate proportional hazards model (Cox) were used to compare prognostic differences between different USP30-AS1 expression groups. The impact of lncRNA USP30-AS1 expression on other genomic analyses was also analyzed. Results High expression of USP30-AS1 was significantly associated with the International Federation of Gynecology and Obstetrics (FIGO) stage of the tumor. Multivariate survival analysis indicated that USP30-AS1 expression level served as an independent prognostic marker for OSC. GSEA data showed that high expression of USP30-AS1 might activate programmed death 1 (PD-1) signaling pathway, cytotoxic T lymphocyte-associated protein 4 (CTLA4) pathway, B-cell receptor signaling pathway, cell apoptosis, fibroblast growth factor receptor (FGFR) signaling pathway, and Janus kinase/signal transducer and activator of transcription (JAK/STAT) signaling pathway. The expression of USP30-AS1 was negatively correlated with immune cell infiltration, including B cells, CD4+ T cells, dendritic cells, CD8+ T cells, and neutrophils. Conclusion USP30-AS1 may be used as a prognostic molecular marker for OSC.
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Female , Humans , Pregnancy , CD8-Positive T-Lymphocytes , Computational Biology , Cystadenocarcinoma, Serous/genetics , RNA, Antisense , RNA, Long Noncoding/genetics , Ubiquitin-Specific Proteases/geneticsABSTRACT
Objective:To observe the repair effect and possible mechanism of Dipsacus saponins Ⅵ on tibial fracture model rats.Methods:Thirty Sprague Dawley (SD) rats were randomly divided into model group, intervention group, and combination group, with 10 rats in each group, to establish a tibial fracture rat model using the sawing method. On the second day after surgery, the intervention group was intraperitoneally injected with 10 mg/kg of Chuanduduan saponin Ⅵ; The combination group received intraperitoneal injection of Dipsacus saponins Ⅵ 10 mg/kg and XAV939 1 mg/animal; The model group was intraperitoneally injected with 0.2 ml of physiological saline solution and 0.2 ml of dimethylsulfoxide (DMSO) solution; Once a day, continuous intervention for 14 days. After 2 to 4 weeks of intervention, Micro CT scan and X-ray scan were used to observe the fracture healing status; After 4 weeks of intervention, the wet weight of the tibia was detected; Hematoxylin eosin (HE) staining was used to observe the pathological changes of callus tissue; The Western blot method was used to detect the expression level of callus tissue β- catenin (β-catenin), p-β-catenin, glycogen synthase kinase 3β (GSK-3 β) and Runt related transcription factor 2 (Runx2) protein.Results:After 2 and 4 weeks of intervention, the bone volume fraction (BV/TV), number of trabeculae (Tb.N), Lane Sandhu score, and callus volume in the intervention group were higher than those in the model group (all P<0.05); After 2 and 4 weeks of intervention, the BV/TV, Tb.N, Lane Sandhu score, and callus volume in the combined group were lower than those in the intervention group (all P<0.05). The wet weight of the tibia in the intervention group was higher than that in the model group at 4 weeks after intervention ( P<0.05); The wet weight of the tibia in the combined group was lower than that in the intervention group ( P<0.05). The HE staining results showed that the model group had fibrous tissue hyperplasia and more bone trabeculae, but the maturity was not high and the thickening was not significant; The intervention group formed more bony callus, with orderly arrangement of bone trabeculae, partially mature, and obvious mineralization, consistent with the direction of stress; The combined group formed more cartilaginous and fibrous callus, with more mineralization at the edge of the cartilaginous callus and the formation of bone trabeculae. Abundant capillaries can be observed in the gaps. The expression level of Runx2 and p-β-catenin/β-catenin protein in callus tissue of the intervention group was higher than that of the model group, the protein expression GSK-3 β level was lower than that of the model group (all P<0.05); The expression level of Runx2 and p-β-catenin/β-catenin protein in the callus tissue of the combined group was lower than that of the intervention group; the protein expression level of GSK-3β was higher than that of the intervention group (all P<0.05). Conclusions:Dipsacus saponins Ⅵ can effectively promote fracture repair in tibial fracture model rats; It is possible to plays a role by activating the Wnt/β-catenin signaling pathway.
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Competing endogenous RNAs (CeRNAs) are crisscrossing regulatory networks. CeRNAs networks can mediate malignant tumor cell phenotypes, including proliferation and inhibition, autophagy, infinite growth, induction of angiogenesis and angiogenic mimicosis, immune escape, etc.. It is expected to provide new diagnostic markers and therapeutic targets for malignant tumors to master the regulation and function of CeRNAs mediated phenotype in malignant tumors.
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Objective To investigate the clinical effects of minimally invasive poking reduction technique in the treatment of single segment thoracolumbar fractures without neural impairment.Methods From February 2011 to June 2015,83 cases of thoracolumbar fractures without neural impairment underwent minimally invasive pedicle screw fixation in Linyi Central Hospital were selected and randomly divided into two groups.Group A (40 cases) was treated with poking reduction technique by percutaneous polyaxial pedical screw fixation,43 patients in group B were treated with only percutaneous polyaxial pedicle screw fixation.The perioperative index,pre-and postoperative radiography,relief of the low back pain and general health status of the two groups were recorded and compared.Results There were no statistically significant differences in the operation time,operative blood loss,hospitalization time.All patients were followed up for 20-27months (average 24 months),the scores of visual analogue scale (VAS) and Oswestry disablity index(ODI) had no statistically significant differences between the two groups in the same period(all P ＞ 0.05).Before operation,the Cobb angle,sagittal index and anterior height of the fracture vertebral body in group A were (66.3 ± 14.2) ％,(20.4 ± 6.5) °,(21.9 ± 6.6) °,respectively,which in group B were (64.8 ± 13.5) ％,(14.5 ± 7.7) °,(15.6 ± 5.9) °,respectively,the differences were not statistically significant (all P ＞ 0.05).After operation,the Cobb angle,sagittal index and anterior height of the fracture vertebral body in group A were (93.8 ± 9.8)％,(5.3 ± 3.3) °,(5.4 ± 2.0) °,respectively,which in group B were (88.0 ± 10.6) ％,(4.1 ± 2.8) °,(8.1 ± 4.7) °,respectively,the differences were statistically significant (t =8.893,2.345,3.351,all P ＜ 0.01).Conclusion The effect of poking reduction technique by percutaneous polyaxial pedical screw fixation is better than simply polyaxial pedicle screw in the treatment of thoracolumbar fracture,which is a safe and effective operation method.
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Objective To explore the expression of RET in cervical squamous carcinoma tissues and evaluate its relationship with cervical squamous carcinoma clinical pathological indexes and its prognostic value. Methods The expression and distribution of RET in normal cervical tissues ,CINⅠ,CINⅡ& CINⅢ and cervical squamous carcinomas tissues were detected by immunohistochemistry in wax blocks. Clinical data and follow-up data are integrated to analyze its relationship with clinical pathological factors and prognosis value. Results The positive rate of RET protein in cervical squamous carcinomas tissues is higher than in other tissues.The positive rate was related to FIGO stage and lymph node metastasis(P0.05). The result of survival analysis with Kaplan-Meier method showed poorer disease-free survival time in the patients with high expression of RET (P < 0.05). Prognostic analysis of patients with cervical cancer through COX proportional hazard regression model suggested that RET expression was an independent prognostic factor. Conclusions RET has been involved in the progression ,FIGO stage and metastasis of cervical squamous cell carcinomas.
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To investigate the expression of metastasis tumor-associated protein 2 (MTA2) in cervical squamous carcinoma and its relationship with prognosis. Methods: Immunohistochemistry and real-time PCR were performed to determine the expression and distribution of MTA2 mRNA and protein in normal cervical tissue, cervical intraepithelial neoplasia (CIN) and cervical squamous carcinomas tissues, then its relationship with clinical pathological factors and prognosis was analyzed. Results: The positive rate of MTA2 protein in normal cervical tissue, CIN and cervical squamous cell carcinomas tissues were 0, 30.0%, 73.4%, respectively. The positive rate was associated with international federation of gynecology and obstetrics (FIGO) stage and lymph node metastasis, whereas there was no correlation with the age of patients or the degree of tumor differentiation. The expression of MTA2 mRNA in normal cervical tissue, CIN and cervical squamous carcinomas tissues was 0.437±0.028, 0.737±0.102 and 1.172±0.068, respectively. The positive rate was associated with FIGO stage and lymph node metastasis, whereas there was no correlation with the age of patients or the degree of tumor differentiation. The result of survival analysis showed poor overall survival time in the patients with high expression of MTA2. Multivariate COX proportional hazards model showed that the positive expression of MTA2 protein, FIGO stage and the metastasis of lymph node were independent prognostic factors for unfavorable clinical outcome of cervical cancer. Conclusion: The positive expression of MTA2 was closely related to the development, invasion and metastasis of cervical squamous cell carcinomas. The positive expression of MTA2 protein, FIGO stage and the metastasis of lymph node were independent prognostic factors for unfavorable clinical outcome of cervical cancer. The expression of MTA2 could be used as a potential molecular marker in evaluating the prognosis of cervical squamous cell carcinomas.
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Female , Humans , Biomarkers, Tumor , Genetics , Carcinoma, Squamous Cell , Genetics , Mortality , Uterine Cervical Dysplasia , Genetics , Gene Expression , Physiology , Histone Deacetylases , Physiology , Immunohistochemistry , Lymph Nodes , Lymphatic Metastasis , Genetics , Neoplasm Staging , Prognosis , Proportional Hazards Models , Real-Time Polymerase Chain Reaction , Repressor Proteins , Physiology , Survival Analysis , Uterine Cervical Neoplasms , Genetics , MortalityABSTRACT
Objective To observe the effect of ephedrine combined propofol and fentanyl in painless induced abortion.Methods Eighty cases of patients (ASA Ⅰ) who apply painless induced abortion were randomly divided into two groups:observation group and control group,each group with 40 cases.Both of the groups were given fentanyl 1 μ g/kg and propofol 2 mg/kg with 2 mg/ml lidocaine; then observation group was given 0.08-0.15 mg/kg ephedrine according to the blood pressure.The heart rate,blood pressure and pulse oximetry (SpO2) of preinduction,3 min and 5 min after induction and 3 min after surgery and the recovery time was observed.Results There was no significant difference about heart rate,blood pressure and SpO2 preinduction,but there was significant difference on 3 min and 5 min after induction in control group in contrast to preinduction [(69.80 ± 7.08),(65.18 ± 5.16) times/min vs.(83.65 ± 8.12)times/min and (86.65 ± 8.60),(90.73 ± 8.35) mmHg (1 mmHg =0.133 kPa) vs.(128.45 ± 11.83) mmHg] (P ＜ 0.05),while observation group kept stable; there was no significant difference about SpO2 and recovery time in both groups.Conclusion It is safe and effective to use ephedrine combined propofol and fentanyl in painless induced abortion.
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Objective To explore the association of expression of apoptosis-associated gene BAK and cFLIP with the biological behaviors in endometriosis. Methods The expression of BAK and cFLIP protein gene in eutopic and ectopic tissue samples from 40 cases with pathologically confirmed ovarian endometriosis and 40 cases with pathologically confirmed normal endometrium was detected by immunohistochemical method. Results ①The expression of BAK and cFLIP protein gene was found in three groups of different endometrial tissue. ②The expression of BAK protein gene was increased gradually in ectopic endometrial , eutopic endometrium and normal tissue and there was significantly difference between every two groups ,while cFLIP was contrary expressed (P 0.05). ④The expression of BAK protein gene in severe group (Ⅲ-Ⅳ period) is lower than mild group both in eutopic or ectopic endometrial tissues,while cFLIP was contrary expressed (P < 0.05). ⑤The expression of BAK and cFLIP was negatively correlated with each other in ectopic endometrium (r=-0.389,P< 0.05). Conclusion BAK and cFLIP was negatively expressed in EMS, which may take a part in the endometrial apoptosis and disorderly proliferation. BAK and cFLIP may play an important role in the the diagnosis and treatment of endometriosis.
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BACKGROUND:Open reduction and internal fixation cause big trauma and many complications. With the progression of minimal y invasive concept, percutaneous pedicle screw fixation gradual y showed its obvious superiority. ＜br> OBJECTIVE:To compare clinical outcomes of minimal y invasive percutaneous pedicle screw fixation versus open surgery in the treatment of thoracolumbar fracture. ＜br> METHODS:From October 2012 to January 2014, 50 cases of thoracolumbar fractures, including 25 cases in the minimal y invasive percutaneous pedicle screw fixation group and 25 cases in the open surgery group, were retrospectively analyzed. The differences in length of skin incision, intraoperative blood loss, operation time, postoperation hospital stay, and visual analog scale scores were compared. Serum creatine kinase activity and C-reactive protein levels were measured before surgery and at 24 and 48 hours after operation. Imaging results were used to observe vertebral height and kyphosis Cobb’s angle changes. ＜br> RESULTS AND CONCLUSION:Compared with the open surgery group, the length of skin incision was smal er and intraoperative blood loss was less, operation time, bed time and hospital stay were shorter, and pain of the wound was lighter in the minimal y invasive group. No significant difference was found in serum creatine kinase activity and C-reactive protein levels between the two groups. Serum creatine kinase activity and C-reactive protein levels were higher at 24 and 48 hours after treatment compared with before treatment in both groups. Serum creatine kinase activity and C-reactive protein levels were higher in the open surgery group than in the minimal y invasive group at 24 and 48 hours. There were significant differences in vertebral height and kyphosis Cobb’s angle in both groups after treatment compared with before treatment (P＜0.01). No significant difference in vertebral height and kyphosis Cobb’s angle was detected between the two groups after treatment (P>0.05). Results indicated that minimal y invasive percutaneous pedicle screw fixation and open surgery in repair of thoracolumbar fractures had similar outcomes. However, the trauma of minimal y invasive percutaneous pedicle screw fixation was apparently less than open surgery.
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Objective To study the expression and significance of human tissue kallikrein gene 6(KLK6) in cervical cancer tissues.Methods With glyceraldehyde 3-phosphate dehydrogenase (GAPDH)as reference,the expression of KLK6 in 80 cases of cervical cancer tissues (40 cases with metastasis and 40cases without metastasis) and 40 cases of normal cervical tissues was determined by Taqman probe real-time quantitative reverse transcription-polymerase chain reaction,and analyzed the relationship between cervical cancer occurring and KLK6 expression with clinical data and pathological dats.Results The expression of KLK6 in normal cervical tissues[(1.06 ± 0.40) × 10-3] was lower than that in cervical cancer tissues without and with metastasis[(4.41 ± 1.70) × 10-3,(32.22 ± 6.70) × 10-3],and there was significant difference (P＜0.01).The expression of KLK6 in Ⅰ a,Ⅰ b,Ⅱ a stage of cervical cancer tissues with metastasis was (30.42 ± 5.00) × 10-3,(31.64 ± 1.30) × 10-3,(33.02 ± 8.00) × 10-3,and there was no significant difference among them (P ＞ 0.05).The expression of KLK6 in Ⅰ a,Ⅰ b,Ⅱ a stage of cervical cancer tissues without metastasis was (4.12 ± 1.10) × 10-3,(4.35 ± 1.30) × 10-3,(4.82 ± 1.90) × 10-3,and there was no significant difference among them (P＞0.05).There was significant difference in the expreesion of KLK6 in Ⅰ a,Ⅰ b,Ⅱ a stage between cervical cancer tissues with metastasis and cervical cancer tissues without metastasis (P ＜0.01).Conclusion KLK6 can stimulate the cervical cancer cell proliferation,and participate in the progresses of cervical cancer metastasis.
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Purpose:To study the effects of arsenic trioxide on the different types of human ovarian cancer cells growth in vitro.Methods:Methyl thiazolyl tetrazolium (MTT)was used to observe the growth inhibition rates of human ovarian cancer cell lines 3AO,SKOV 3 and TYK cells by various concentration arsenic trioxide at different times; Cell apoptosis percentage and cell cycles phase distribution of 3AO were measured by flow cytometry(FCM) assays; Apoptotic phenotype of SKOV 3 was observed by acridine dying. Results:Arsenic trioxide could inhibit the growth of 3AO?SKOV 3 and TYK effectively, depended on the action time and concentration of the medicine(P0.05). Within a certain range, 3AO cells apoptotic percentage induced by arsenic trioxide were enhanced in concentration- and time-dependent patterns(P