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.

OBJECT IVE To provide reference for the improvement of the quality standard for Niushanggui capsules. METHODS Based on the previous quality standard ，thin-layer chromatography （TLC）identification methods were established for Angelicae dahurica and Anemarrhenae asphodeloides . High performance liquid chromatography （HPLC）method was established to determine the contents of 5 components simultaneously ，such as mangiferin ，prim-O-glucosylcimifugin，naringin，neohesperidin and 5-O-methylvisammioside. The limits were confirmed. RESULTS TLC chromatogram of Niushanggui capsules showed the same color spots in the same position as the corresponding （mixed）substance control or reference medicinal material of A. dahuricae and A. asphodeloides ，while the negative samples had no interference. The linear range of mangiferin ，prim-O-glucosylcimifugin， naringin，neohesperidin and 5-O-methylvisammioside were 7.98-127.63，6.74-107.84，53.06-848.96，39.31-628.90，13.54-216.62 μg/mL，respectively（all r＝0.999 9）. RSDs of precision ，stability（24 h）and repeatability tests were all no more than 1.20％（n＝ 6）. The average recoveries were 95.00％，105.16％，97.16％，101.00％ and 104.97％（RSD≤1.50％，n＝6）. In 4 batches of samples，the average contents of the above 5 components were 0.842，0.696，6.951，5.755 and 1.106 mg/g respectively ；the limits of A. asphodeloides ，Saposhnikovia divaricata and Citrus aurantium were based on the contents of mangiferin ，the total content of prim-O-glucosylcimmifugin and 5-O-methylvisammioside，naringin and neohesperidin ，which would not be less than 0.42，0.90 and 6.36 mg/grain，respectively. CONCLUSIONS TLC identification methods of A. dahurica and A. asphodeloides and the content determination methods of 5 components as mangiferin in Niushanggui capsules are established in this study ，and the limits of A. asphodeloides ，S. divaricata and C. aurantium are confirmed.

Objective:To compare Jefferson-fracture reduction plate (JeRP) and micro titanium plate in the transoral single-segment fixation of unstable atlas fractures.Methods:From January 2008 to December 2020, 45 patients with unstable atlas fracture were treated by single-segment fixation through an oral approach with a JeRP or a micro titanium plate at Department of Orthopedic Surgery, General Hospital of Southern Theatre Command. They were 24 males and 21 females, aged from 15 to 67 years. By the Gehweiler classification, 11 atlas fractures were type Ⅰ and 34 type Ⅲ; by the American Spinal Injury Association (ASIA) classification, the spinal cord injury was grade D in 7 cases and grade E in 38 cases; by the Dickman classification, the atlas transverse ligament injury was type Ⅰ in 4 cases and type Ⅱ in 11 cases. Of the patients, 26 were treated by transoral single-segment fixation with a JeRP and 19 by transoral single-segment fixation with a micro titanium plate. The 2 groups were compared in terms of baseline data, operation time, blood loss, hospital stay, visual analog scale (VAS) for neck pain and atlas lateral mass displacement (LMD) before operation and at the last follow-up, and intraoperative and postoperative complications.Results:The 2 groups were comparable because there was no significant difference between them in the preoperative general data ( P>0.05). All patients were followed up for 12 to 55 months (mean, 21.8 months). Wound dehiscence or infection was observed in none of the patients after operation. About 12 months after operation, all fractures achieved bony union, neck pain basically disappeared, and neck movement had no obvious limitation. The hospital stay was (13.9±2.2) d for the JeRP group and (14.2±2.9) d for the micro titanium plate group, showing no significant difference between the 2 groups ( P>0.05). The operation time was (203.5±173.4) min and the blood loss (167.3±138.6) mL in the JeRP group, significantly more than those in the micro titanium plate group [(121.5±50.5) min and (98.4±57.2) mL] ( P<0.05). In the JeRP group, the preoperative LMD was (6.7±1.7) mm and the preoperative VAS score (6.8±1.0) points, significantly higher than the last follow-up values [(0.7±0.6) mm and (0.7±0.6) points] ( P<0.05). In the micro titanium plate group, the preoperative LMD was (6.6±1.5) mm and the preoperative VAS score (6.7±0.9) points, significantly higher than the last follow-up values [(0.9±0.6) mm and (0.8±0.7) points] ( P<0.05). However, there was no significant difference in the preoperative or the last follow-up comparison between the 2 groups ( P>0.05). Implant loosening was observed in one patient in the JeRP group while foreign body sensation in the throat was reported in one patient after operation in the micro titanium plate group. Conclusions:Both JeRP and micro titanium plate in the transoral single-segment fixation can lead to effective treatment of unstable atlas fractures. Compared with JeRP, the micro titanium plate can effectively shorten operation time and reduce blood loss due to its smaller size and lower incision.

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: To explore the effects of various forms of prostatic apex on positive apical margin rate (PAM) and biochemical recurrence (BCR) after laparoscopic radical prostatectomy. Methods: A retrospective analysis of 309 patients (aging (65±6) years) who were experienced laparoscopic radical prostatectomy from January 2010 to December 2016 at the Department of Urology, First Affiliated Hospital of Fujian Medical University. According to the relationship between prostate apex and membrane urethra at the mid-sagittal plane of preoperative MRI, all patients were classified into 4 categories. There were 31 patients for type 1, apex covering both anterior and posterior aspects of membranous urethra, 139 patients for type 2, apex covering anterior side of membranous urethra, 63 patients for type 3, apex covering posterior aspect of membranous urethra, 76 patients for type 4, apex not covering membranous urethra. PAM and BCR after operation were compared between this four groups respectively. The χ² test was used to compare PAM among the 4 types. Logistic regression analysis were undertaken to analyze the factors affecting PAM. Cox′s proportional hazards regression model was undertaken to identify the variables influencing BCR. Results: There was no significant difference in the 4 groups concerning age, body mass index, prostate volume, preoperative prostate-specific antigen (PSA) value, postoperative Gleason score and pathological stage (P>0.05).The median follow-up time was 32 months (ranged from 12 to 60 months).The data showed that the apical type 3 patients has the highest PAM. There was statistical difference among the 4 groups in PAM (χ²=15.592, P=0.001). Preoperative level of PSA (OR=20.356, 95% CI: 2.440 to 169.810, P=0.005), postoperative Gleason score (OR=4.113, 95% CI: 1.911 to 8.849, P=0.001), pathological stage (OR=3.422, 95% CI: 1.600 to 7.319, P=0.002) and apical type 3 (OR=6.134, 95% CI: 2.196 to 17.132, P=0.001) were independent relactive factors of PAM. Preoperative level of PSA (HR=1.362, 95% CI: 1.006 to 1.843, P=0.045), postoperative Gleason score (HR=1.920, 95% CI: 1.384 to 2.665, P=0.001), pathological stage (HR=1.476, 95% CI: 1.098 to 1.983, P=0.010), PAM (HR=3.497, 95% CI: 2.407 to 5.081, P=0.001)and apical type 3 (HR=1.828, 95% CI: 1.266 to 2.639, P=0.001) were independent prognosis factors of BCR. Conclusion Prostate apical type 3 could be a significant independent predictor of PAM, and an independent prognosis factor for BCR.

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.