Objective To investigate the influence of classically activated macrophage (M1) on the proliferation of rheumatoid arthritis (RA) fibroblast-like synovial (FLS) and osteoarthritis (OA) FLS proliferation.Methods Human monocytes leukemia cells (THP)-1 were induced into M1 by lipopolysaccharides (LPS) and interferon gamma (IFN-γ),M1 specific surface molecular markers human leukocyte antigen (HLA)-DR and CD197 were detected by flow cytometry (FCM).RA-FLS and OA-FLS were co-cultured with M1 by transwell chambers,the proliferation of RA-FLS and OA-FLS were observed by crystal violet staining assay.MTS was used to detect cytokines secreted from M1 on the multiplication of RA-FLS and OA-FLS.TNF-α and IL-12 were detected by enzyme linked immunosorbent assay (ELISA).Paried student t test was used for statistical analysis.Results THP-1 were induced into M1 by LPS and IFN-γ,the expression rates of M1 surface specific molecular markers HLA-DR and CD197 were 78.25％ and 87.96％.Crystal violet staining showed that RA-FLS and OA-FLS proliferation were significantly inhibited after co-cultured with M1 48 h,RA-FLS and OA-FLS of each vision under microscope in co-culture groups were (64 ±30) and (85 ±23) respectively,while the RA-FLS and OA-FLS in separate culture groups were (467±87) and (263±78) respectively,the difference was statistically significant (t=7.459,3.791;P＜0.05).MTS assay indirectly reflected that the cytokines from M1 suppressed RA-FLS and OA-FLS proliferation (t=-7.155,-8.111;P＜0.05).The concentration of TNF-α in cell culture supernatants secreted from RA-FLS group and RA-FLS/M1 co-culture group respectively were (0.024±0.01 1) ng/ml and (0.832±0.241) ng/ml respectively,the concentration of IL-12 from the two groups were (0.033±0.015) ng/ml and (0.372±0.122) ng/ml respectively.TNF-α from OA-FLS and OA-FLS/M1 co-culture group respectively were (0.031±0.017) ng/ml and (0.852±0.323) ng/ml,IL-12 were (0.012±0.009) ng/ml,(0.373±0.144) ng/ml.Compared with FLS separate culture group,the concentration of TNF-α and IL-12 were obviously elevated (t=-4.997,-4.777,-4.407,-4.334;P were all ＜0.05).Conclusion M1 can significantly inhibite RA-FLS and OA-FLS proliferation,this may be related to the increased concentration of TNF-α and IL-1 β in from cell culture supernatant.

Objective This study was performed to investigate the effect of hypoxia on glucose-6-phosphate isomerase (G6PI) expression and cell cycle of fibroblast-like synoviocytes from synovium of rheumatoid arthritis (RA) and osteoarthritis (OA) under hypoxia or normoxia.Methods Fibroblast-like synoviocytes were cultured with either of hypoxia (3％ oxygen) or normoxia (21％ oxygen) for 24 hours.The mRNA expression of G6PI and HIF-1α was tested by PCR quantification,while the protein levels of G6PI and HIF-1α were measured by western blot.Cell cycle was performed by FACS.T-test and Mann-Whitney U were used for statistical analysis.Results The expression levels of G6PI mRNA under hypoxia in RA were higher than those of OA (2.6±0.4 vs 1.5±0.4,P＜0.05).The protein levels of G6PI in RA were higher than those of OA (P＜0.05).The expression levels of HIF-1α mRNA under hypoxia in RA were higher than those of OA (2.9±0.8vs 1.4 ±0.4,P＜0.05).The protein levels of HIF-1α in RA were higher than those of OA (P＜0.05).The G1 phase ratio of cell cycle was decreased significantly under hypoxia than those of normoxia in RA ELs (t=1 1.31,P＜0.05).The S and G2 phase ratio of cell cycle were increased.Conclusion Hypoxia upregulates G6PI and HIF-1α expression and improves proliferation in fibroblast-like synoviocytes.

r study.

Objective The current study is aimed to investigate the association of single nucleotide polymorphisms and gene expression in peptidylarginine deiminase Ⅳ (PADI4) with rheumatoid arthritis (RA).Methods The gene expression of PADI4 was examined using real-time quantitative reverse transcription-polymcrase chain reaction (RT-PCR) in 70 patients with RA and 81 healthy controls. Four exonie SNPs of the PADI4 gene (PADI4_89, _90, _92, _104) were genotyped using DNA sequencing and TA cloning.Results The distribution of PADI4_89, _90, _104 SNPs in RA was different from that of healthy controls. The increased RA susceptibility was significantly associated with minor alleles. When haplotypes were construe -ted with 4 SNPs, two major haplotypes, ACCC and GTGT were found in all samples, and GTGT haplo-type (carrying only the minor alleles) was significandy associated with increased RA susceptibility (P<0.01)in comparison with the reference haplotype ACCC. There was over expression of PADI4 in RA than controls(P<0.05). C, enotypes carrying the minor alleles had higher expression level of PADI4 in RA and controls than those with the common alleles. Conclusion PADI4 SNPs and haplotypes are associated with RA susceptibility in Chinese. PADI4 is over-expressed in the blood of RA patients, and there is significant association between the haplotypes and expression level of the PADI4 gene.

Objective:To prove the effect of PADI-4 gene in the development of rheumatoid arthritis.Methods:Four SiRNA sequences were designed for PADI-4 gene,and the SiRNAs were cloned into blank pSiRNA-hH1neo G2 vectors.The vectors were transformed into GT116 E.coli competent cells.By white-blue selection system,the right vectors were gotten.After transfection into HL-60 cells,the cells were collected on 3,5,7,10 and 14 day,the levels of PADI-4 mRNA were detected by Real-time PCR.Results:Digestion by Acc 65Ⅰand Hind Ⅲ,the recombinant expressive vector of RNA interference was obtained successfully.The PADI-4 mRNA generated by the cells transfected with the vector of SiRNAs were reduced,and the level was not change in normal cells.Conclusion:The recombinant expressive vector of RNA interference is obtained successfully and the recombinant expressive vector can affect expression of PADI-4 gene in HL-60 cells.

Objective: To evaluate the effect of multimodal analgesia using periprostatic nerve block anesthesia (PNB) combined with flurbiprofen in patients undergoing transperineal template-guided prostate biopsy (TTPB). Methods: Totally 166 patients (aged (68.2±9.1) years, range: 47 to 81 years) who received TTPB from October 2017 to June 2018 at Department of Urology, Northern Jiangsu People′s Hospital Affiliated to Yangzhou University were enrolled prospectively. All the patients were randomly divided into 2 groups. The observation group (n=79) was given flurbiprofen axetil 1 mg/kg intravenously for half an hour before operation and lidocaine was used for PNB before the biopsy. The control group (n=87) was given normal saline combined with PNB. A visual analog scale (VAS) and visual numeric scale (VNS) were used to assess the patients′ pain and quantify their satisfaction at two time points: VAS-1 and VNS-1: during biopsy procedure, VAS-2 and VNS-2: 30 min after the procedure. The date were compared by t test, χ² test, Fisher exact test and two-way repeated measures anova analysis between the 2 groups. Results: The age, total prostate volume, serum prostate-specific antigen and the number of cores were comparable among the 2 groups (P>0.05). The VAS-1 scores of the control group and the observation group were 2.8±1.7, 1.9±1.2, respectively, and the VNS-1 were 3.1±0.7, 3.4±0.3, respectively. The VAS-1 were significantly lower in observation group than in control group (F=3.904, P=0.000). Conversely, the VNS-1 were higher in observation group (F=3.526, P=0.000). At 30-minute postoperative, the VAS-2 and VNS-2 were 0.7±0.4 and 3.7±0.2 in the control group, respectively. The VAS-2 and VNS-2 were 0.6±0.5 and 3.8±0.1 in the observation group, respectively. There were no significant differences in the pain scores or the satisfaction scores between the 2 groups (F=1.429, 2.825; P=0.136, 0.083). The incidence of overall complications was 26.4% (23/87) in the control group and 25.3% (20/79) in the observation group, with no statistical difference between the 2 groups (χ²=0.027, P=0.869). And the complications had no statistically significant difference among the 2 groups including hematuria, urinary retention, infection, hematospermia, vascular and neurological reactions, nausea, vomiting, dizziness, headache, and respiratory depression (P>0.05). Conclusion The multimodal analgesia induced by PNB and flurbiprofen could effectively relieve the pain for patients who received TTPB.

Objective:To explore the feasibility and safety of the clinical application of the diagnosis and treatment mode combining rapid frozen pathological examination of prostate biopsy tissue with radical prostatectomy.Methods:Suspected prostate cancer patients with PSA>10 ng/ml and PI-RADS score ≥4 in, Northern Jiangsu People's Hospital from April to September 2021 were collected. The included patients underwent mpMRI/TRUS image fusion-guided transperineal prostate targeted biopsy with 16G biopsy needle, 2-3 needles for biopsy, and rapid frozen pathological examination. Robot-assisted laparoscopic radical prostatectomy (RALP) was performed immediately for patients with prostate cancer with rapid freezing pathology. For undiagnosed prostate cancer, 18G biopsy needle for prostate targeted + systematic biopsy were used, 18-22 needles for systematic biopsy, and routine pathological examination. The baseline data, frozen pathological results, perioperative conditions, pathological results and follow-up data of all patients were collected.Results:Eleven patients were included in the study, the mean age of the patients was 69.9(66-73) years, the mean BMI was 22.8(19-26) kg/m 2, the mean PSA was 23.2(14.25-32.00), the mean prostate volume was 45(32-52) ml, mean PSAD 0.54(0.33-0.75). PI-RADS score was 4 in 3 cases and 5 in 8 cases; digital rectal examination was positive in 5 cases. All 11 cases underwent rapid freezing and the pathological results showed that: 9 cases were prostate adenocarcinoma, and RALP was performed immediately. The operation time was 111.5(96-126) min, the intraoperative blood loss was 78.9(55-105) ml, and the postoperative extubation time was 4.3(3.5-5.0) days, postoperative hospital stay 5.8(5.0-6.5) days. Postoperative pathology showed that Gleason score 3+ 4=7 in 1 case, 4+ 3=7 in 3 cases, 8 points in 4 cases, and 10 points in 1 case; 3 cases had positive resection margins, and 1 case had seminal vesicle invasion, the average number of dissected lymph nodes was 10.9 (8.5-14.0), and there was no tumor metastasis. Pathological T staging included 2 cases of T 2b stage, 5 cases of T 2c stage, 1 case of T 3a stage, and 1 case of T 3b stage. Two patients were undiagnosed by rapid freezing pathology, of which one was prostate adenocarcinoma with a Gleason score of 4+ 3=7, and then received RALP; the other one was prostate inflammation. 11 patients were followed up; the postoperative follow-up time was 3-7 months, with an average of 5.2 months. Among the 10 patients who underwent RALP, 8 patients recovered urinary continence 2 weeks after surgery, and all recovered within 2 months after surgery. Three patients with positive surgical margins were given regular androgen deprivation therapy in the second week after surgery. PSA did not drop below 0.1 ng/ml in patients with positive margins and seminal vesicle invasion 3 months after surgery. No complications of Clavien grade Ⅰ or higher occurred after operation and during follow-up. Conclusions:For patients with high suspicion of prostate cancer, rapid frozen pathological examination of prostate biopsy tissue is performed. RALP is performed immediately for patients with prostate cancer. The results show that this diagnosis and treatment model could be safe and feasible.

Objective:To investigate the effect of local anesthesia in patients with a PI-RADS score of 5 and ECOG score ≥2 for prostate puncture.Methods:Retrospective analysis of case data of 33 patients admitted to the Subei People's Hospital for prostate puncture from April 2020 to April 2022. Age (82.5±3.6) years. There were 18 cases with hypertensive disease, 8 cases with diabetes mellitus, and 6 cases with both diabetes mellitus and hypertensive disease. Body mass index (25.2±3.5) kg/m 2. prostate-specific antigen (PSA)(131.5±69.7) ng/ml. prostate volume (38.5±21.4) ml. all patients had a PI-RADS score of 5 on multiparametric magnetic resonance (mpMRI) and an Eastern Cooperative Oncology Group (ECOG) score ≥2. All 33 cases in this group underwent trans-perineal targeted prostate puncture using local anesthesia at the tip of the prostate. The visual analog score (VAS) and visual numeric score (VNS) were applied by the same surgeon to assess the patient's pain level and satisfaction at the time of puncture (VAS-1 and VNS-1) and 30 min after puncture (VAS-2 and VNS-2), and to record the duration of the procedure and the occurrence of postoperative complications. Results:In this group of 33 cases, the VAS-1 score was (1.9±0.3) and the VAS-2 score was (0.1±0.2); the VNS-1 score was (2.9±0.2) and the VNS-2 score was (3.9±0.1). Postoperative pathological results indicated that one of the 33 patients had a negative puncture result (pathology report indicating interstitial inflammation), while the rest of the patients had a positive puncture pathology report (puncture pathology report indicating prostate cancer), with a positive rate of 97%. One case of postoperative carnal haematuria occurred, which gradually improved after the patient was advised to drink water and take alpha-blockers. No perineal hematoma occurred, and all patients did not suffer complications such as urinary tract infection, urinary retention, azoospermia, vagal reaction, and infectious shock.Conclusion:In patients with a PI-RADS score of 5 and ECOG score ≥2, the use of single-hole local anesthesia for performing trans-perineal targeted puncture biopsy has the advantages of good paroxysmal pain and high safety.

Objective To evaluate the clinical efficacy of brachytherapy with 125I seeds implantation in patients with castration resistant prostate cancer (CRPC).Methods Twenty-eight patients with CRPC from February 2010 to December 2015 in Northern Jiangsu People's Hospital were analyzed retrospectively.Patients were divided into 2 groups according to different treatment methods.Control group (n =13) received endocrine therapy,and study group (n =15) underwent brachytherapy combined with endocrine therapy.The progression was defined as posttreatment prostate specific antigen (PSA) ≥ 125％ PSA of baseline level.PSA-progression-free survival (PFS),overall survival (OS) and quality of life (physical functioning,social functioning,general health,general physical discomfort,urinary symptoms and treatment-related symptoms) of 2 groups were compared using Kaplan-Meier analysis and log-rank test.Results The median PSA-PFS and OS of study group were 29(24,37) months and 37(32,50) months respectively,both of them were longer than those of control group (13(7,21) months,19(14,23) months;x2 =13.300,19.362,both P＜0.01).Comparing with control group,the physical functioning,social functioning,general health and general physical discomfort of study group were improved.Conclusion Brachytherapy can effectively prolong the survival of CRPC patients and improve the patients' life quality.

Objective:To analyze the risk of missed diagnosis in patients with PI-RADS score>3 and negative prostate initial biopsy and to explore its risk factors.Methods:The clinical data of 268 patients with negative prostate biopsy in Northern Jiangsu People's Hospital from May 2013 to December 2018 were retrospectively analyzed. The patients were divided into observation group (PI-RADS score>3) and control group (PI-RADS score≤ 3) according to different PI-RADS scores. There were insignificant differences in age [(67.4(60.0, 74.0)years and 65.6(66.5, 72.0)years], prostate volume of initial biopsy [62.4(40.0, 72.0)ml and 60.8(38.0, 77.0)ml], biopsy cores [ 20.6(18.0, 22.0)cores and 20.4(18.0, 22.0)cores] between the observation group (n=124) and the control group(n=144)(all P>0.05). But there were significant differences in PSA [17.5(6.5, 23.0)ng/ml and 11.5(6.3, 12.0)ng/ml], PSAD[0.316(0.128, 0.363)ng/ml 2 and 0.211(0.106, 0.256)ng/ml 2], prostate inflammation of the initial biopsy [70 (56.5%) and 32 (22.2%)] between the observation group and the control group(all P<0.05). According to the follow-up results after the initial biopsy, the two groups of repeated biopsy were compared.Furthermore, Logistic regression was used to conduct univariate and multivariate analysis to explore the risk factors of patients with PI-RADS>3 for positive repeated biopsy. At the same time, the receiver operating characteristic curve (ROC curve) was used to analyze the accuracy of the risk factors. Results:There were significant differences in repeated biopsy rate [ 27.4%(34/124)and 14.6%(21/144)], CsPCa detection rate[ 41.4%(14/34) and 4.8%(1/21)]between the observation group and the control group(all P<0.05). The positive rate of repeated biopsy in the observation group (41.1%) was higher than that in the control group (23.8%), but there was no statistical difference ( P=0.248). The risk of positive repeated biopsies in the observation group was 2.24 times than that in the control group. Univariate analysis found repeated biopsy PSA ( P =0.02, OR=1.438, 95% CI 1.161-1.896), PSA ratio (repeated biopsy PSA/initial biopsy PSA) ( P=0.011, OR=10.087, 95% CI 1.714-59.36) were risk factors for positive of repeated biopsy in patients with PI-RADS score >3. Multivariate analysis also found that repeated biopsy PSA ( P=0.017, OR=1.15, 95% CI 1.076-2.123), PSA ratio ( P=0.032, OR=10.2, 95% CI 0.883-116.168) were risk factors for positive repeated biopsy. ROC curve analysis, the accuracy of repeated biopsy PSA (AUC=0.971, P<0.001, 95% CI 0.926-1.000), PSA ratio (AUC=0.839, P=0.001, 95% CI0.707-0.971) to predict positive of repeated biopsy were high. The cut-off values were 21.3 ng/ml and 1.4, respectively. The accuracy was higher when combines repeated biopsy PSA with PSA ratio (AUC=0.993, P<0.001, 95% CI 0.974-1.000). Conclusions:Patients with negative PI-RADS score > 3 have a higher risk of missed diagnosis of CsPCa than those with PI-RADS score≤3. When PSA>21.3 ng/ml and PSA ratio>1.4 during follow-up, the possibility of missed diagnosis in the initial biopsy is high.