Membranous nephropathy (MN) is an autoimmune disease in the kidney glomerulus and one of the leading causes of nephrotic syndrome. It can be characterized by the regular immune complexes deposited between the podocyte and the glomerular basement membrane, causing nephrotic syndrome in adults. Early diagnosis and treatment of MN are of great importance. MN can be divided into idiopathic membranous nephropathy (IMN) and secondary membranous nephropathy (SMN) according to its etiology. Anti-M-type phospholipase A2 receptor (PLA2R) antibodies, anti-platelet reaction-protein type 1 7A domain (THSD7A) antibodies have been used for MN diagnosis. Since THSD7A, many target antigens such as neural epidermal growth factor-like 1 protein (NELL1), semaphorin 3B (Sema3B), protocadherins7 (PCDH7), serine protease HTRA1 (HTRA1), exostosin 1/exostosin 2 (EXT1/EXT2), and neural cell adhesion molecule1 (NCAM1) have been identified as MN-related biomarkers. The newly identified antigens are of unique clinical values, especially for PLA2R and THSD7A negative MN diagnosis.

Objective:To report our experience in diagnosis and treatment of posterior atlantoaxial dislocation with odontoid retrolisthesis.Methods:A retrospective study was conducted of the 5 patients who had been treated from July 2012 to August 2018 at Department of Orthopaedics, General Hospital of Southern Theater Command for posterior atlantoaxial dislocation. They were 4 men and one woman, aged from 34 to 67 years (average, 47 years). All of them had a history of trauma. Of them, 4 were complicated with odontoid fracture and one with congenital free os odontoideum. Their posterior atlantoaxial dislocation ranged from 3 to 9 mm (average, 6 mm). By the American Spinal Injury Association (ASIA) grading system, their preoperative spinal injury was rated as grade B in one, as grade C in 3 cases and as grade D in one. All the 5 patients underwent skull traction at 10° flexion. Surgical trans-oralpharyngeal atlantoaxial reduction and internal fixation was performed for the one patient whose reduction had not been achieved by traction while posterior atlantoaxial screw-rod fixation or anterior odontoid screwing was conducted for the 4 patients whose reduction had been achieved by traction. The distance of posterior atlantoaxial dislocation was measured to evaluate their reduction and ASIA grade system was used to assess their spinal function after operation.Results:The postoperative distance of posterior atlantoaxial dislocation was 0 mm, showing a reduction rate of 100%. The 5 patients were followed up for 6 to 36 months (average, 15 months). By the ASIA grade system, the postoperative functional recovery of the spine was grade D in 4 cases and grade C in one. No implant loosening or breakage occurred.Conclusion:As a kind of high-energy hyperextension injury, posterior atlantoaxial dislocation is rare in clinic, but an appropriate treatment can be adopted to deal with its different clinical types to achieve good outcomes.

Objective To evaluate the predictors of gleason score pathological downgrading after radical prostatectomy in patients with biopsy-proven level 2 of grading groups (Gleason Score 3 + 4 =7).Methods Data of 252 patients,diagnosed with level 2 of grading groups(Gleason score 3 + 4 =7) prostate cancer by biopsy,with subsequent laparoscopic radical prostatectomy,were retrospectively analyzed.The mean age was 64.3,ranged from 46 to 82 years.The average body mass index (BMI) was 23.2 kg/m2,ranged from 15.2 to 30.4 kg/m2.The median prostate volume,transition zone volume(TZV) and transition zone index(TZI) were 48.9 ml (30.3-73.1 ml),21.4 ml(13.5-31.2 ml) and 0.46％ (0.37％-0.58％),respectively.The median value of tPSA,fPSA and PSAD were 1.53 ng/ml(0.67-3.92 ng/ml),9.65 ng/ml (4.13-18.68 ng/ml) and 0.18 ng/(ml · cm3) [0.09-0.50 ng/ (ml · cm3)],respectively.Clinical T stage was also evaluated,including 153 (60.7％) diagnosed as T1e stage,78 (3 1.0％) diagnosed as T2 stage,and 21 (8.3％) diagnosed as T3 stage.There were 58(23.0％) with extracapsular extension,47 (18.7％) patients with seminal vesicle invasion,and 2(0.8％) with lymph node metastasis.Pathological T stage includes 112 (44.4％) diagnosed as T2 stage,55 (21.8％) diagnosed as T3a stage,35 (13.9％) diagnosed as T3b stage,and 50(19.8％) diagnosed as T4 stage.The patients were assigned Prostate ImagingReporting and Data System version 2 scores of 1,2,3,4,and 5 were 45 (17.9％),36 (14.3％),51 (20.2％),68(27.0％)and 52(20.6％),respectively.The patients were categorized into 2 groups with and without pathological downgrading,including downgrade and no downgrade group.Univariate and multivariate logistic regression analysis were done to determine the predictors of pathological downgrading.Results The patients were categorized into downgrade(n =31) and no downgrade group(n =221) of 252 patients.The pathological downgrading was identified in 31 (12.3％).The tPSA,PSAD and PI-RADS scores in patients with downgrade group which were lower than those in without downgrade group (P ＜ 0.05).The logistic regression analysis revealed PI-RADS score was the independent predictor of downgrading(OR =0.364,95％ CI 0.253-0.522,P ＜ 0.01).The area under the ROC curve of PI-RADS score was 0.810 and the diagnostic value was the best.Conclusions These findings suggested that PI-RADS scores was predictor for pathological downgrading after radical prostatectomy in patients with biopsy-proven level 2 of grading groups,reduced PI-RADS score (PI-RADS score ≤ 3) is correlated with increased pathological downgrading after radical prostatectomy.

Objective Detecting plasma level of circular RNA(circRNA)hsa_circ_0009024 in pulmonary tuberculosis(TB)patients,and evaluating its diagnostic value for TB.Methods From January 2016 to December 2016, a hosptial-based, case-control study was performed, which include 90 untreated active pulmonary tuberculosis patients(TB group),75 healthy people(healthy control)and 84 patient with other diseases(other disease group).Plasma level of circRNA hsa_circ_0009024 was detected by real-time quantitative polymerase chain reaction.Furthermore, the 90 patients with TB were divided into different subgroups according to cavity formation and the lung fields involvement: patients without lung cavity(55 cases)vs those with lung cavity(35 cases),patients with involvement of <2 lung fields(49 cases)vs≥2 lung fields(41 cases).Plasma levels of hsa _circ_0009024 of 41 TB patientswere monitored andcomparedbefore and after 3 months anti-TB therapy.The sensitivity and specificity of plasma hsa_circ_0009024 were analyzed by using the receiver operating characteristic(ROC)curve.The comparison between two groups was performed with Mann-Whitney U test and the comparison among multigroupswas conducted with Kruskal-Wallis H test.Results Plasma levels of hsa_circ_0009024 in TB patients[1.98(1.42, 2.71)]were significantly higher than healthy controls[1.03(0.78,1.33)]and other disease groups[1. 13(0.77,1.51)](H=76.58,P<0.0001).Plasma levels of hsa_circ_0009024 in cavity pulmonary TB patients were higher than pulmonary TB patients without cavity(U=392.50,P<0.0001).Plasma levels of hsa_circ_0009024 in TB patients with involvement of ≥2 lung fields were higher than <2 lung fields(U=590.50,P=0.0008).As compared to pre -treatment[2.01(1.41, 2.71)], the plasma hsa_circ_0009024 levels decreased significantly in 3 months[1.22(0.85,1.47)](U=292.50,P<0.0001)after anti-TB therapy.The area under the receiver operating characteristics curve(AUC)of plasma hsa_circ_0009024 in discriminating the patients with TB from normal controls, pneumonia patients and lung cancer patients were 0.841 and 0.811, respectively.Conclusion The hsa_circ_0009024 can be used as a potential biomarker in TB diagnosis and monitoring.

AIM:To investigate the role of Rho-associated kinase ( ROCK) inhibitor fasudil in the formation of rabbit urethral stricture after injury and to observe the cell activity , migration and extracellular matrix synthesis in the rabbit urethra fibroblasts.METHODS:The rabbit model of urethral stricture was established by microsurgical techniques .The rabbits were divided into sham operation group , operation group and fasudil (3 mg/kg, 10 mg/kg, 30 mg/kg) groups.The diameter of the stenosis was measured by retrograde urethrography 3 months after surgery .The fibroblasts were isolated from urethral scar, and then incubated with fasudil (12.5 μmol/L, 25 μmol/L, 50 μmol/L) in the presence of transforming growth factor-β1 (TGF-β1, 10 μg/L).The untreated cells were used for control .The cell activity was measured by MTT assay.The cell migration ability was tested by the method of Transwell chambers .The protein expression of ROCK , α-smooth muscle actin (α-SMA) , collagen I and collagen III was determined by Western blot analysis .RESULTS:Fasudil significantly reduced formation of urethral stricture after injury (P<0.05).Cultured rabbit fibroblasts with different con-centrations of fasudil inhibited the cell activity and cell migration ability (P<0.05).The protein expression of ROCK,α-SMA, collagen I and collagen III was also inhibited by treatment with fasudil in a dose -dependent manner ( P<0.05 ) . CONCLUSION:Fasudil inhibits the formation of extracellular matrix and reduces the incidence of urethral stricture after injury by down-regulating TGF-β1-induced Rho/ROCK pathway activation in the rabbit urethra fibroblasts .

OBJECTIVETo investigate the role of glycogen synthase kinase 3β (GSK-3β) in the maturation and function of murine bone marrow-derived dendritic cells (BMDCs).

METHODSMature DCs (mDCs) induced by LPS were examined for GSK-3β phosphorylation level with Western blotting before and after LPS exposure. To explore the role of GSK-3β in maturation and function of DCs, we added SB216763, a selective inhibitor of GSK-3β, in the cell culture of immature DCs (iDCs), and examined CD40 and CD86 expressions in the cells by flow cytometry and the expression of IL-6, IL-12 and IL-10 mRNA by real-time PCR; the changes of the immunogenicity of the cells was evaluated by mixed lymphocyte reaction. The expression of GSK-3β and RelB was examined by Western blotting in DC2.4 cells transfected with a lentiviral vector over-expressing murine GSK-3β gene.

RESULTSLPS exposure significantly lowered GSK-3β activity in iDCs as demonstrated by increased Ser9 phosphorylation and reduced Tyr216 phosphorylation. GSK-3β inhibition induced DC maturation by increasing the expression of surface costimulatory molecules CD40 and CD86, lowered the expressions of IL-6 and IL-12 while enhanced the expression of IL-10 in iDCs, and impaired mixed lymphocyte reaction of the cells. In DC2.4 cells, lentivirus-mediated over-expression of GSK-3β obviously down-regulated the expression of RelB.

CONCLUSIONSGSK-3β is a crucial enzyme involved in the differentiation and maintenance of an immature phenotype of DCs. GSK-3β is constitutively active in iDCs to inhibit their spontaneous maturation. DCs become phenotypically mature after inhibition of GSK-3β, which also executes a proinflammatory task in DC activation. The reduction of RelB protein levels as a result of GSK-3β overexpression supports GSK-3β as a new target for inducing tolerogenic DCs.

Animals ; B7-2 Antigen ; metabolism ; CD40 Antigens ; metabolism ; Cell Differentiation ; Cells, Cultured ; Culture Media ; chemistry ; Dendritic Cells ; enzymology ; Glycogen Synthase Kinase 3 ; metabolism ; Glycogen Synthase Kinase 3 beta ; Indoles ; chemistry ; Interleukin-10 ; metabolism ; Interleukin-12 ; metabolism ; Interleukin-6 ; metabolism ; Lentivirus ; Lymphocyte Culture Test, Mixed ; Maleimides ; chemistry ; Mice ; Myeloid Cells ; enzymology ; Phosphorylation ; RNA, Messenger ; Real-Time Polymerase Chain Reaction ; Signal Transduction

Objective To investigate the role of glycogen synthase kinase 3β(GSK-3β) in the maturation and function of murine bone marrow-derived dendritic cells (BMDCs). Methods Mature DCs (mDCs) induced by LPS were examined for GSK-3βphosphorylation level with Western blotting before and after LPS exposure. To explore the role of GSK-3βin maturation and function of DCs, we added SB216763, a selective inhibitor of GSK-3β, in the cell culture of immature DCs (iDCs), and examined CD40 and CD86 expressions in the cells by flow cytometry and the expression of IL-6, IL-12 and IL-10 mRNA by real-time PCR;the changes of the immunogenicity of the cells was evaluated by mixed lymphocyte reaction. The expression of GSK-3βand RelB was examined by Western blotting in DC2.4 cells transfected with a lentiviral vector over-expressing murine GSK-3βgene. Results LPS exposure significantly lowered GSK-3βactivity in iDCs as demonstrated by increased Ser9 phosphorylation and reduced Tyr216 phosphorylation. GSK-3β inhibition induced DC maturation by increasing the expression of surface costimulatory molecules CD40 and CD86, lowered the expressions of IL-6 and IL-12 while enhanced the expression of IL-10 in iDCs, and impaired mixed lymphocyte reaction of the cells. In DC2.4 cells, lentivirus-mediated over-expression of GSK-3βobviously down-regulated the expression of RelB. Conclusion GSK-3βis a crucial enzyme involved in the differentiation and maintenance of an immature phenotype of DCs. GSK-3β is constitutively active in iDCs to inhibit their spontaneous maturation. DCs become phenotypically mature after inhibition of GSK-3β, which also executes a proinflammatory task in DC activation. The reduction of RelB protein levels as a result of GSK-3β overexpression supports GSK-3β as a new target for inducing tolerogenic DCs.

Objective To investigate the role of glycogen synthase kinase 3β(GSK-3β) in the maturation and function of murine bone marrow-derived dendritic cells (BMDCs). Methods Mature DCs (mDCs) induced by LPS were examined for GSK-3βphosphorylation level with Western blotting before and after LPS exposure. To explore the role of GSK-3βin maturation and function of DCs, we added SB216763, a selective inhibitor of GSK-3β, in the cell culture of immature DCs (iDCs), and examined CD40 and CD86 expressions in the cells by flow cytometry and the expression of IL-6, IL-12 and IL-10 mRNA by real-time PCR;the changes of the immunogenicity of the cells was evaluated by mixed lymphocyte reaction. The expression of GSK-3βand RelB was examined by Western blotting in DC2.4 cells transfected with a lentiviral vector over-expressing murine GSK-3βgene. Results LPS exposure significantly lowered GSK-3βactivity in iDCs as demonstrated by increased Ser9 phosphorylation and reduced Tyr216 phosphorylation. GSK-3β inhibition induced DC maturation by increasing the expression of surface costimulatory molecules CD40 and CD86, lowered the expressions of IL-6 and IL-12 while enhanced the expression of IL-10 in iDCs, and impaired mixed lymphocyte reaction of the cells. In DC2.4 cells, lentivirus-mediated over-expression of GSK-3βobviously down-regulated the expression of RelB. Conclusion GSK-3βis a crucial enzyme involved in the differentiation and maintenance of an immature phenotype of DCs. GSK-3β is constitutively active in iDCs to inhibit their spontaneous maturation. DCs become phenotypically mature after inhibition of GSK-3β, which also executes a proinflammatory task in DC activation. The reduction of RelB protein levels as a result of GSK-3β overexpression supports GSK-3β as a new target for inducing tolerogenic DCs.

OBJECTIVETo explore the effect of different functional groups on self-assembled monolayers on the biological characteristics of rabbit skeletal muscle cells in vitro.

METHODSRabbit skeletal muscle cells were cultured on self-assembled monolayers of gold on which different terminal chemical groups including methyl groups (-CH(3)), amino(-NH(2)), hydroxyl(-OH) and carboxyl (-COOH ) were anchored with self-assembled methods. Contact angle measurements and atomic force microscopy were employed to confirm the similar density of different functional groups occupation. Fluorescence microscopy, MTT assay, flow cytometry, and scanning electron microscopy (SEM) were used to analyze the morphological and biological alterations of the cells.

RESULTSSEM results revealed that the chemical groups on the surface of the monolayer modulated the structure of skeletal muscle cells and the cell morphology. Skeletal muscle cells cultured on the monolayer with -CH3 exhibited the smallest contact area with a spherical morphology, while the cells on the monolayers with -NH(2), -OH and -COOH showed much larger contact area and flatter morphology. The functional groups -NH(2) and -COOH obviously promoted cell adhesion and proliferation, while -CH(3) group produced significantly greater toxicity than -NH(2), -OH and -COOH groups to inhibit the cell growth and adhesion and promote cell death. Cell attachment and growth was enhanced, in the order the magnitude of the effect, by -NH(2)>-COOH>-OH>-CH(3), and the toxicity decreased in the order of -NH(2)>-COOH>-OH>-CH(3).

CONCLUSIONThe terminal chemical groups can obviously affect the phenotype of skeletal muscle cells in vitro, and this finding provides a theoretical basis for surface design of biomaterials.

Animals ; Cell Adhesion ; Cell Proliferation ; Cells, Cultured ; Microscopy, Atomic Force ; Microscopy, Electron, Scanning ; Microscopy, Fluorescence ; Muscle Fibers, Skeletal ; cytology ; Rabbits

Objective To study the expression of reversion-inducing-cysteine-rich protein with Kazal motifs (RECK) in human pancreatic carcinoma tissues and pancreatic carcinoma cell lines; the effects of recombinant lentiviruses carrying RECK gene(LV-RECK) therapy on human pancreatic carcinoma xenograft in nude mice; and to find out the relationship between the expression of RECK and the prognosis of pancreatic carcinoma.Methods Immunohistochemical method was used to detect the expression of RECK in the resected specimens of pancreatic carcinoma and their corresponding normal pancreatic tissues in 42 patients.Western blotting was used to examine the expression of RECK in human pancreatic carcinoma cell lines (PANC-1,MIAPaCa-2,AsPC-1).Statistical analyses were performed to determine the relationship between RECK expression and the clinicopathological characteristics and prognosis in pancreatic carcinoma.Subcutaneous xenograft tumor models of human pancreatic carcinoma were established in nude mice.These nude mice were then divided into the experimental group,the negative control group and the blank control group randomly.The three groups of nude mice were intratumorally injected with LV-RECK,LV-EGFP and normal saline (NS) respectively.The antitumor effect was studied.Immunohistochemical method was used to detect the expression of RECK and microvessel density (MVD).Terminal deoxynucleotidyl transferase mediated dUTP-DIG nick end labeling (TUNEL) was used to detect the apoptosis of tumor cells.Survival analysis was performed.Results All three pancreatic carcinoma cell lines did not express RECK.The overall positive rate of RECK expression was 45.2 ％ (19/42) in pancreatic carcinoma,and 88.1 ％ (37/42) in normal pancreatic tissue.The expression level of RECK was significantly lower in the tumor tissues than in the normal tissues (P＜0.01).The expression of RECK was significantly associated with TNM stage,lymph node metastasis and local infiltration of pancreatic carcinoma (P＜0.05).Kaplan-Meier survival analysis revealed that the survival time was significantly longer in the RECK positive patient group than in the RECK negative patient group.Univariate Cox regression analysis revealed that RECK expression,TNM stage,lymph node metastasis and local infiltration were significantly related with prognosis for pancreatic carcinoma (P＜0.05).Multivariate Cox regression analysis revealed that only RECK expression remained as an independent significant factor in predicting the prognosis of pancreatic carcinoma (P ＜ 0.001).When compared with the negative control and the blank control groups,the volume of subcutaneous xenograft tumor in the experimental group was significantly decreased (P＜0.05).RECK protein in the experimental group was re-expressed.MVD of the experimental group was significantly less than those of the control groups (P＜0.05).Apoptotic index (AI)of the experimental group was significantly higher than those of the control groups (P＜0.05).The survival time of nude mice in the experimental group was significantly longer than those in the control groups (P＜0.05).Conclusions RECK expression was closely related to invasion,metastasis and prognosis of pancreatic carcinoma and it was an independent prognostic marker.RECK gene over-expression inhibited neovascularization of pancreatic carcinoma,induced apoptosis of tumor cells,inhibited the growth of tumor xenograft and improved the prognosis of tumor-bearing mice.These results suggest a possible new treatment for pancreatic carcinoma.