d-Galactose induces the senescence and phenotype switch of corpus cavernosum smooth muscle cells.

Studies regarding age-related erectile dysfunction (ED) based on naturally aging models are limited by their high costs, especially for the acquisition of primary cells from the corpus cavernosum. Herein, d-galactose ( d-gal) was employed to accelerate cell senescence, and the underlying mechanism was explored. As predominant functional cells involved in the erectile response, corpus cavernosum smooth muscle cells (CCSMCs) were isolated from 2-month-old rats. Following this, d-gal was introduced to induce cell senescence, which was verified via ß-galactosidase staining. The effects of d-gal on CCSMCs were evaluated by terminal deoxynucleoitidyl transferase dUTP nick-end labeling (TUNEL), immunofluorescence staining, flow cytometry, western blot, and quantitative real-time polymerase chain reaction (qRT-PCR). Furthermore, RNA interference (RNAi) was carried out for rescue experiments. Subsequently, the influence of senescence on the corpus cavernosum was determined via scanning electron microscopy, qRT-PCR, immunohistochemistry, TUNEL, and Masson stainings. The results revealed that the accelerated senescence of CCSMCs was promoted by d-gal. Simultaneously, smooth muscle alpha-actin (alpha-SMA) expression was inhibited, while that of osteopontin (OPN) and Krüppel-like factor 4 (KLF4), as well as fibrotic and apoptotic levels, were elevated. After knocking down KLF4 expression in d-gal-induced CCSMCs by RNAi, the expression level of cellular alpha-SMA increased. Contrastingly, the OPN expression, apoptotic and fibrotic levels declined. In addition, cellular senescence acquired partial remission. Accordingly, in the aged corpus cavernosum, the fibrotic and apoptotic rates were increased, followed by downregulation in the expression of alpha-SMA and the concurrent upregulation in the expression of OPN and KLF4. Overall, our results signaled that d-gal-induced accelerated senescence of CCSMCs could trigger fibrosis, apoptosis and phenotypic switch to the synthetic state, potentially attributed to the upregulation of KLF4 expression, which may be a multipotential therapeutic target of age-related ED.

Effects of Degreasing Pretreatment on Immunohistochemistry and Molecular Analysis of Gastrointestinal and Breast Cancer Samples.

Lymph node status is a key factor in determining stage, treatment, and prognosis in cancers. Small lymph nodes in fat-rich gastrointestinal and breast cancer specimens are easily missed in conventional sampling methods. This study examined the effectiveness of the degreasing pretreatment with dimethyl sulfoxide (DMSO) in lymph node detection and its impact on the analysis of clinical treatment-related proteins and molecules. Thirty-three cases of gastrointestinal cancer specimens from radical gastrectomy and 63 cases of breast cancer specimens from modified radical mastectomy were included. After routine sampling of lymph nodes, the specimens were immersed in DMSO for 30 minutes for defatting. We assessed changes in the number of detected lymph nodes and pN staging in 33 gastrointestinal cancer specimens and 37 breast cancer specimens. In addition, we analyzed histologic characteristics, Masson trichrome special staining, and immunohistochemistry (gastrointestinal cancer: MMR, HER2, and PD-L1; breast cancer: ER, PR, AR, HER2, Ki-67, and PD-L1). Molecular status was evaluated for colorectal cancer (KRAS, NRAS, BRAF, and microsatellite instability) and breast cancer (HER2) in gastrointestinal cancer specimens and the remaining 26 breast cancer specimens. Compared with conventional sampling, DMSO pretreatment increased the detection rate of small lymph nodes (gastrointestinal cancer: P < .001; breast cancer: P < .001) and improved pN staging in 1 case each of gastric cancer, colon cancer, and rectal cancer (3/33; 9.1%). No significant difference in the morphology, special staining, protein, and molecular status of cancer tissue after DMSO treatment was found. Based on these results and our institutional experience, we recommend incorporating DMSO degreasing pretreatment into clinical pathologic sampling practices.

A systematic scoping review of communication skills training in medical schools between 2000 and 2020.

BACKGROUND: Communication skills training (CST) remains poorly represented and prioritised in medical schools despite its importance. A systematic scoping review (SSR) of CST is proposed to better appreciate current variability in their structure, content, and assessment. This is to guide their future design in medical school curricula. METHODS: The Systematic Evidence-Based Approach (SEBA) was used to guide concurrent SSRs of teaching and assessment in CST. After independent database searches, concurrent thematic and content analysis of included articles were conducted separately. Resultant themes/categories were combined via the jigsaw perspective to provide a more holistic view of the data. These were then compared to tabulated summaries of the included articles to create funnelled domains. RESULTS: 52,300 papers were identified, 150 full-text articles included, and four funnelled domains were identified: Indications, Design, Assessment, and Barriers and Enablers of CST. CSTs confer numerous benefits to physicians and patients. It saw increased confidence, improved diagnostic capabilities and better clinical management, as well as greater patient satisfaction and treatment compliance. Skills may be divided into core, prerequisite competencies, and advanced skills pertinent to more challenging and nuanced scenarios - such as population or setting-specific situations. CST teaching and assessment modalities were found to align with Miller's Pyramid, with didactic teaching gradually infused with experiential approaches to enhance their understanding and integration. A plethora of CST frameworks, teaching and assessment methods were identified and are presented together. CONCLUSION: While variable in approach, content and assessment, CST in medical schools often employ stage-based curricula to instil competency-based topics of increasing complexity throughout medical school education. This process builds on the application of prior knowledge and skills, influencing practice and, potentially, the students' professional identity formation. In addition, the institution plays a critical role in overseeing training, ensuring longitudinal guidance and holistic assessments of the students' progress.

MicroRNA-20a Targeting LASS2 Promotes the Proliferation, Invasiveness and Migration of Bladder Cancer.

BACKGROUND: Abnormal expression of miR 20a is reported in various types of malignancy neoplasms. However, its function is not consistent in different tumors. This study aims to explore the potential functions of miR 20a and its underlying mechanisms in bladder cancer. METHODS: Ninety-six patients diagnosed with bladder cancer were recruited into the study. The expression levels of miR-20a in bladder cancer samples and adjacent non-tumor samples were investigated by qRT-PCR. Wound healing, CCK8, and transwell migration assays were carried out for determining the functions of miR20a. Bioinformatics analysis was used for predicting the downstream gene of miR-20a. Western blot, qRT-PCR, and fluorescent reporter assays were used to verify the target gene. RESULTS: MiR-20a was significantly increased in bladder cancer tissues, and its rising level was closely correlated with histological grade, clinical stage, recurrence and metastasis in bladder cancer. Exogenous upregulation of miR-20a expression obviously enhanced the aggressive biological functions of bladder cancer in vitro. LASS2 was verified to be a target gene of miR-20a. Moreover, miR-20a can negatively regulate LASS2 at protein and mRNA levels. CONCLUSIONS: Increasing miR-20a is closely related to aggressive clinicopathological features. MiR 20a plays an oncogenic role in bladder cancer, which contributes to target LASS2 directly.

Super-Mini Percutaneous Nephrolithotomy Reduces the Incidence of Postoperative Adverse Events in Pediatric Patients: A Retrospective Cohort Study.

OBJECTIVE: To assess the safety of the super-mini percutaneous nephrolithotomy (SMP) versus the minimally invasive percutaneous nephrolithotomy (MPCNL) in the treatment of pediatric renal calculus. METHODS: We retrospectively reviewed the electronic records of pediatric patients who underwent treatment for renal stones by either SMP or MPCNL from May 2015 to May 2016. We compared the safety of the 2 surgical procedures in the treatment of renal calculus in children by using the generalized estimating equation (GEE) multivariate regression analysis, in which the exposures are the surgical procedures and postoperative adverse events (postoperative complications, fever, and WBC counts) are set as outcome variables. RESULTS: The study included 39 patients (26 boys and 13 girls), of which 22 underwent MPCNL and 17 underwent SMP, with a mean age of 110.05 ± 45.01 and 93.18 ± 41.72 months, respectively. In the univariate logistic regression model, the surgical procedures showed no significant association with postoperative complications (95% CI 0.0-1.5), fever (95% CI 0.1-2.1), postoperative peripheral WBC (95% CI 0.1-2.2). In the multiple logistic regression analysis, there was an insignificant association between surgical methods and postoperative complications (95% CI 0.28-1.1), fever (95% CI 0.1-1.2), and postoperative peripheral WBC (95% CI 0.03-1.8). While using GEE with multiple dependent variables and MPCNL as a reference, the OR of adverse events was 0.15 and the 95% CI were 0.04-0.55. CONCLUSIONS: Compared to MPCNL, SMP has a lower incidence of postoperative complications and appears to be a safer treatment for children with kidney stones.

Cry1 deficiency leads to testicular dysfunction and altered expression of genes involved in cell communication, chromatin reorganization, spermatogenesis, and immune response in mouse testis.

Cryptochrome (Cry)1 is essential for generating circadian rhythm in central and many peripheral oscillators; however, its role in male reproduction remains unclear. We investigated this question using Cry1 knockout (KO) mice. We found that Cry1 is necessary for normal testicular function: Cry1 deficiency increased testicular germ cell apoptosis and decreased sperm count. A transcriptome analysis showed that the expression levels of 375 genes-including 12 encoding micro (mi)RNAs-were altered in the testis of Cry1 KO mice relative to wild-type controls. A bioinformatics analysis revealed that the differentially expressed genes were related to important biological processes including cell-cell communication, metabolism, chromatin reorganization, spermatogenesis, and the immune response. An integrative analysis of miRNA-mRNA networks suggested that the 12 dysregulated miRNAs may contribute to testis disorders through negative regulation of their target mRNAs expression in testis, and interestingly, all the 12 miRNAs are predicted to target core circadian clock components. These results provide the first evidence of a correlation between dysregulation of Cry1 and male reproductive defects in mice, indicating that Cry1 plays a critical role in maintaining normal testicular function.

Cryo-EM structure of cyanophage P-SCSP1u offers insights into DNA gating and evolution of T7-like viruses.

Cyanophages, together with their host cyanobacteria, play important roles in marine biogeochemical cycles and control of marine food webs. The recently identified MPP-C (Marine Picocyanobacteria Podovirus clade C) cyanophages, belonging to the T7-like podoviruses, contain the smallest genomes among cyanopodoviruses and exhibit distinct infection kinetics. However, understanding of the MPP-C cyanophage infection process is hindered by the lack of high-resolution structural information. Here, we report the cryo-EM structure of the cyanophage P-SCSP1u, a representative member of the MPP-C phages, in its native form at near-atomic resolution, which reveals the assembly mechanism of the capsid and molecular interaction of the portal-tail complex. Structural comparison of the capsid proteins of P-SCSP1u and other podoviruses with known structures provides insights into the evolution of T7-like viruses. Furthermore, our study provides the near-atomic resolution structure of portal-tail complex for T7-like viruses. On the basis of previously reported structures of phage T7, we identify an additional valve and gate to explain the DNA gating mechanism for the T7-like viruses.

Primary angiomatoid fibrous histiocytoma of the mandible with EWSR1-ATF1 fusion in an adult patient: case report and review of literature.

OBJECTIVE: We report our diagnosis of a rare case of primary angiomatoid fibrous histiocytoma in the mandible of a 42-year-old male using next-generation sequencing to detect disease-specific EWSR1-ATF1 fusion. STUDY DESIGN: After the initial cone beam computerized tomography scan and reconstruction, we performed immunohistochemical staining and fluorescence in situ hybridization analysis on tissue samples to detect EWSR1 gene rearrangement. For the final diagnosis, we performed next-generation sequencing to detect disease-specific EWSR1-ATF1 fusion. RESULTS: FISH analysis showed approximately 55% of tumor cells with mostly isolated red signals, as well as several split red-green signals, indicating the presence of EWSR1 gene rearrangement. Next-generation sequencing analysis identified an EWSR1 exon9-ATF1 exon4 fusion, a diagnostic biomarker of angiomatoid fibrous histiocytoma (AFH). Based on the findings, we diagnosed primary AFH derived from the mandible. CONCLUSIONS: Next-generation sequencing is a powerful methodology for detecting disease-specific EWSR1-ATF1 fusion and diagnosing primary angiomatoid fibrous histiocytoma.

Abundant and cosmopolitan lineage of cyanopodoviruses lacking a DNA polymerase gene.

Cyanopodoviruses affect the mortality and population dynamics of the unicellular picocyanobacteria Prochlorococcus and Synechococcus, the dominant primary producers in the oceans. Known cyanopodoviruses all contain the DNA polymerase gene (DNA pol) that is important for phage DNA replication and widely used in field quantification and diversity studies. However, we isolated 18 cyanopodoviruses without identifiable DNA pol. They form a new MPP-C clade that was separated from the existing MPP-A, MPP-B, and P-RSP2 clades. The MPP-C phages have the smallest genomes (37.3-37.9 kb) among sequenced cyanophages, and show longer latent periods than the MPP-B phages. Metagenomic reads of both clades are highly abundant in surface waters, but the MPP-C phages show higher relative abundance in surface waters than in deeper waters, while MPP-B phages have higher relative abundance in deeper waters. Our study reveals that cyanophages with distinct genomic contents and infection kinetics can exhibit different depth profiles in the oceans.

Tumor-infiltrating lymphocytes status, programmed death-ligand 1 expression, and clinicopathological features of 41 cases of pure apocrine carcinoma of the breast: a retrospective study based on clinical pathological analysis and different immune statuses.

Background: Pure apocrine carcinoma (AC) of the breast can be divided into human epidermal growth factor receptor-2 (HER2)-positive and triple-negative apocrine carcinoma (TNAC). Some studies showed that triple negative breast cancer with low tumor-infiltrating lymphocytes (TILs) and high programmed death-ligand 1 (PD-L1) status may be a therapeutic target for immune checkpoint inhibitors. However, the clinicopathological features of different HER2 expression, TILs status and PD-L1 expression in AC are not clear. Therefore, we investigate the status of TILs and PD-L1, as well as the clinicopathological features of pure apocrine carcinoma of the breast. Methods: We retrospectively analyzed the clinicopathological data, and prognosis of 41 cases of pure apocrine carcinoma of the breast that underwent surgical resection from January 2014 to November 2020. TILs were evaluated. Immunohistochemistry (IHC) staining was applied to detect PD-L1 protein expression in 14 of these samples from January 2019 to November 2020. The expression and correlation of HER2, TILs, PD-L1 and clinicopathological features and prognoses were analyzed. Results: A total of 80.5% (33/41) of patients had TILs <50%, and 19.5% (8/41) had TILs ≥50%. The expression of TILs and the Ki-67 proliferation index were significantly higher in the HER2-positive group (41.5%, 17/41) compared to the HER2-negative group (58.5%, 24/41) (P<0.05). Approximately 52.9% (9/17) of HER2-positive patients treated with Trastuzumab targeted therapy, overall survival was higher in HER2-positive patients than in HER2-negative patients (P=0.211). The PD-L1 positivity rate was 50% (7/14) in the 14 pure apocrine carcinoma of the breast samples, and 66.7% (4/6) and 37.5% (3/8) in the HER2-negative and HER2-positive groups, respectively, with no significant difference (P=0.592). Among these 14 cases, two samples had TILs ≥50%, both of which were positive for PD-L1 and Ki67 >20%; and 12 cases had TILs <50%, of which 41.7% (5/12) were PD-L1-positive and 58.3% (7/12) were PD-L1-negative. All 14 cases with PD-L1-negative had TILs <50%. There was no significant difference in overall survival between TILs and Ki67 co-expression (P=0.452). Conclusions: Pure AC HER2-positive patients have higher levels of TILs and Ki67, HER2 negative and TILs ≥50% patients may have higher PD-L1 expression, which may be helpful for screening patients with different immune statuses to guide effective clinical treatment combinations.

Combined transcriptomic and proteomic analysis reveals multiple pathways involved in self-pollen tube development and the potential roles of FviYABBY1 in self-incompatibility in Fragaria viridis.

Fragaria viridis exhibits S-RNase-based gametophytic self-incompatibility, in which S-RNase is the major factor inhibiting pollen tube growth. However, the pathways involved in and the immediate causes of the inhibition of pollen tube growth remain unknown. Here, interactive RNA sequencing and proteome analysis revealed changes in the transcriptomic and proteomic profiles of F. viridis styles harvested at 0 and 24 h after self-pollination. A total of 2,181 differentially expressed genes and 200 differentially abundant proteins were identified during the pollen development stage of self-pollination. Differentially expressed genes and differentially abundant proteins associated with self-incompatible pollination were further mined, and multiple pathways were found to be involved. Interestingly, the expression pattern of the transcription factor FviYABBY1, which is linked to polar growth, differed from those of other genes within the same family. Specifically, FviYABBY1 expression was extremely high in pollen, and its expression trend in self-pollinated styles was consistent with that of S-RNase. Furthermore, FviYABBY1 interacted with S-RNase in a non-S haplotype way. Therefore, FviYABBY1 affects the expression of polar growth-related genes in self-pollen tubes and is positively regulated by S-RNase.

Long Noncoding RNA HAGLROS Promotes the Malignant Progression of Bladder Cancer by Regulating the miR-330-5p/SPRR1B Axis.

Bladder cancer (BC) is the most common genitourinary malignancy worldwide, and its aetiology and pathogenesis remain unclear. Accumulating evidence has shown that HAGLROS is closely related to the occurrence and progression of various cancers. However, the biological functions and underlying mechanisms of HAGLROS in BC remain unknown. In the present study, the expression of HAGLROS in BC was determined by public dataset analysis, transcriptome sequencing analysis, qRT-PCR and ISH assays. Gain- or loss-of-function assays were performed to study the biological roles of HAGLROS in BC cells and nude mouse xenograft model. Bioinformatic analysis, qRT-PCR, western blot, immunohistochemistry, FISH assays, subcellular fractionation assays and luciferase reporter assays were performed to explore the underlying molecular mechanisms of HAGLROS in BC. Here, we found that HAGLROS expression is significantly upregulated in BC tissues and cells, and elevated HAGLROS expression was related to higher pathologic grade and advanced clinical stage, which is significant for BC diagnosis. HAGLROS can enhance the growth and metastasis of BC in vitro and in vivo. Furthermore, miR-330-5p downregulation reversed the BC cells proliferation, migration and invasion inhibited by silencing HAGLROS. SPRR1B silencing restored the malignant phenotypes of BC cells promoted by miR-330--5p inhibitor. Mechanistically, we found that HAGLROS functions as a microRNA sponge to positively regulate SPRR1B expression by sponging miR-330-5p. Together, these results demonstrate that HAGLROS plays an oncogenic role and may serve as a potential biomarker for the diagnosis and treatment of BC.

Laboratory Management and Quality Control Practice of SARS-CoV-2 Nucleic Acid Detection.

OBJECTIVE: A positive result of SARS-CoV-2 nucleic acid detection provides critical laboratory evidence for clinical confirmed diagnosis, pandemic status evaluation, a pandemic prevention plan, treatment of infected people with symptoms, and protection of uninfected people. This study aims to provide a practical reference for SARS-CoV-2 nucleic acid-related research and detection. METHODS: Our laboratory has established policies combining personnel management and quality control practices for SARS-CoV-2 nucleic acid detection during the pandemic. RESULTS: In this article, we describe cross-department personnel management and key points of personal protection and quality control in the testing process. We also report on the differences in detection and the compatibility between different brand kits. CONCLUSION: It is critical to maintain a standard and accurate laboratory operation for nucleic acid testing.

Development of ELISA and Lateral Flow Immunoassays for Ochratoxins (OTA and OTB) Detection Based on Monoclonal Antibody.

Ochratoxins were important secondary metabolites secreted by fungi, and OTA and OTB are mainly significant mycotoxin, having toxic effects on humans and animals. Therefore, it is important to establish a rapid, sensitive, and precise method for ochratoxins detection and quantification in real samples. In this study, a stable monoclonal antibody (mAb) that recognizing both OTA and OTB toxins was employed for the establishment of indirect competitive ELISA (ic-ELISA), colloidal gold nanoparticles (CGNs), and nanoflowers gold strips (AuNFs) for detection of ochratoxins in real samples. A 6E5 hybridoma cell line stable secreting mAb against both OTA and OTB toxins was obtained by fusion of splenocytes with myeloma SP2/0 cells. The 6E5 mAb had a high affinity (3.7 × 108 L/mol) to OTA, and also showed similar binding activity to OTB. The optimized ic-ELISA resulted in a linear range of 0.06-0.6 ng/mL for ochratoxins (OTA and OTB) detection. The IC50 was 0.2 ng/mL and the limit of detection (LOD) was 0.03 ng/mL. The mean recovery rate from the spiked samples was 89.315 ± 2.257%, with a coefficient variation of 2.182%. The result from lateral flow immunoassays indicated that the LOD of CGNs and AuNFs were 5 and 1 µg/mL, respectively. All these results indicated that the developed ic-ELISA, CGNs, and AuNFs in this study could be used for the analysis of the residual of ochratoxins (OTA and OTB) in food and agricultural products.

The Pivotal Role of Host Organizations in Enhancing Mentoring in Internal Medicine: A Scoping Review.

In undergraduate and postgraduate medical education, mentoring offers personalized training and plays a key role in continuing medical education and the professional development of healthcare professionals. However, poor structuring of the mentoring process has been attributed to failings of the host organization and, as such, we have conducted a scoping review on the role of the host organization in mentoring programs. Guided by Levac et al's methodological framework and a combination of thematic and content analysis, this scoping review identifies their "defining" and secondary roles. Whilst the "defining" role of the host is to set standards, nurture, and oversee the mentoring processes and relationships, the secondary roles comprise of supporting patient care and specific responsibilities toward the mentee, mentor, program, and organization itself. Critically, striking a balance between structure and flexibility within the program is important to ensure consistency in the mentoring approach whilst accounting for the changing needs and goals of the mentees and mentors.

Volume-Enhanced Compatible Remeshing of 3D Models.

Compatible remeshing provides meshes with common connectivity structures. The existing compatible remeshing methods usually suffer from high computational cost or poor quality. In this paper, we present a fast method for computing compatible meshes with high quality. Given two closed, oriented, and topologically equivalent surfaces and a sparse set of corresponding landmarks, we first compute a bijective inter-surface mapping, from which compatible meshes are generated. We then improve the remeshing quality by using a volume-enhanced optimization. In contrast to previous work, our method designs a fast volume-enhanced improvement procedure that directly reduces the isometric distortion of the map between the compatible meshes. Our method also tries to preserve the shapes of the input meshes by projecting the vertices of the compatible meshes onto the input surfaces. Central to this approach is the use of the monotone preconditioned conjugate gradient method, which minimizes the energies effectively and efficiently. Compared with state-of-the-art methods, our method performs about one order of magnitude faster with better remeshing quality. We demonstrate the efficiency and efficacy of our method using various model pairs.

Preparation of Monoclonal Antibody for Brevetoxin 1 and Development of Ic-ELISA and Colloidal Gold Strip to Detect Brevetoxin 1.

Brevetoxin-1 (BTX-1), a marine toxin mostly produced by the dinoflagellatae Karenia brevis, has caused the death of marine organisms and has had numerous toxicological effects on human health. Hence, it is very necessary to develop a rapid, economical, and reliable immunoassay method for BTX-1 detection. In this study, two kinds of complete antigen were synthesized using the succinic anhydride and isobutyl chloroformate two-step methods. Conjugate BTX-1-OVA was used as an antigen for mice immunization, and BTX-1-BSA for measuring the titer of the produced antibodies. A hybridoma cell line 6C6 stably secreting monoclonal antibody (mAb) against BTX-1 was obtained by fusing SP2/0 myeloma cells with the spleen cells from the immunized mouse. The hybridoma 6C6 was injected into the abdomen of BALB/c mice to obtain ascites, and the anti-BTX-1 mAb was harvested from ascites by precipitation with caprylic acid/ammonium sulfate (CA-AS). The anti-BTX-1 mAb was identified as an IgG1 subtype, and the cross-reactivity results showed that anti-BTX-1 mAb was highly specific to BTX-1 with the affinity of 1.06 × 108 L/mol. The indirect competitive ELISA results indicated that the linear range for BTX-1 detection was 14-263 ng/mL with IC50 of 60 ng/mL, and a detection limit of 14 ng/mL. The average recovery rate from the spiked samples was 88 ± 2% in intra-assay and 89 ± 2% in inter-assay. The limit of detection (LOD) using the colloidal gold strip was 200 ng/mL with high specificity. Therefore, the anti-BTX-1 mAb can be used to detect BTX-1 in shellfish and other related samples.

Preparation of Anti-Human Podoplanin Monoclonal Antibody and its application in Immunohistochemical Diagnosis.

Podoplanin (PDPN), a 38 kDa transmembrane sialoglycoprotein from human, is expressed in lymphatic endothelial cells but not in vascular endothelial cells, and has been considered as a specific marker of lymph. In this study, the gene encoding the extracellular part of PDPN (ePDPN) was synthesized and used to expressed fusion protein ePDPN-His and GST-ePDPN, respectively, in E.coli. The purified GST-ePDPN fusion protein was mixed with QuickAntibody-Mouse5W adjuvant to immune mice, and the antiserum titer was determined by indirect ELISA. A stable cell line named 5B3 generating anti-PDPN monoclonal antibody (mAb) was obtained by hybridoma technology. The isotype of 5B3 cell line was IgG2b, and the chromosome number was 102 ± 4. The 5B3 mAb was purified successfully from ascites fluid through Protein G column, and its affinity constant was 2.94 × 108 L/mol. Besides, excellent specificity of the 5B3 mAb was further demonstrated in ELISA, western blot and immunohistochemistry experiments, suggesting that 5B3 mAb displays similar application value to D2-40, a commercial available antibody. Hence, the current study provides conclusive guidelines for preparation of other mAbs and their applications in immunohistochemistry diagnosis.

SOCS6 Functions as a Tumor Suppressor by Inducing Apoptosis and Inhibiting Angiogenesis in Human Prostate Cancer.

BACKGROUND: Our previous studies revealed that the downregulation of Suppressor of cytokine signaling 6 (SOCS6) was correlated with malignant progression of human prostate cancer (PCa). AIMS: In the current study, we aimed to investigate the tumor suppressive roles of SOCS6 and the underlying mechanisms in PCa. METHODS: SOCS6 expression in PCa and non-cancerous prostate tissues was compared by immunohistochemistry. Statistical associations of SOCS6 expression with various clinicopathological features and patients prognosis were evaluated. In addition, we investigated SOCS6's functions by overexpressing it in vitro (cell apoptosis, migration and invasion assays) and in vivo (tumor formation, angiogenesis and apoptosis). Moreover, SOCS6-regulated genes were identified by nextgeneration RNA-sequencing analysis, followed by pathway enrichment analysis and in vitro experimental validation. RESULTS: SOCS6 downregulation was significantly associated with advanced clinical stage (P=0.029) and positive lymph node metastasis (P=0.013) in PCa patients. We also identified SOCS6 as an independent prognostic factor for disease-free survival in PCa patients (P=0.045). Moreover, overexpression of SOCS6 inhibited PCa cell invasion, migration, tumor xenografts growth and angiogenesis, but induced PCa cell apoptosis (P values <0.05). Mechanically, we revealed that SOCS6 expression may induce cell apoptosis coincident with down-regulation of Bcl2 and Hspa1a, and may suppress tumor angiogenesis with downregulation of F7, Fak3 and Frzb. CONCLUSION: These findings suggest that the reduced expression of SOCS6 may be predictive of unfavorable prognosis in PCa. Thus, SOCS6 may serve as a tumor suppressor and a novel therapeutic target for this cancer.

Randomized Study of Ureteral Catheter vs Double-J Stent in Tubeless Minimally Invasive Percutaneous Nephrolithotomy Patients.

OBJECTIVE: To prospectively analyze and compare the outcomes of using externalized ureteral catheter (EUC) vs Double-J ureteral stent (DJ) in tubeless minimally invasive percutaneous nephrolithotomy (MPCNL). PATIENTS AND METHODS: A total of 109 patients underwent tubeless MPCNL in our institute and have been enrolled into this study. Fifty-six and 53 patients had EUC and DJ positioning at the conclusion of the procedure, respectively. The two approaches have been compared for operative time, intraoperative blood loss, postoperative visual analogue pain scale (VAS) score, analgesic requirement, stent-related symptoms, hospital stay, degree of vesicoureteral reflux (VUR) on the operative side, and complications according to the modified Clavien system. RESULTS: There were no statistically significant differences between the two groups regarding the mean operative times, mean VAS scores, analgesic requirements, mean hemoglobin drop, mean hospital stay, and overall complication rate. However, compared with DJ group, EUC group presented fewer postoperative stent-related symptoms and less occurrence of severe VUR (p < 0.05). CONCLUSION: Positioning EUC in tubeless MPCNL is a safe alternative to DJ in patients with renal or upper ureteral calculi. EUC provides several benefits: obviated the need of a second endoscopic procedure, reduced stent-related discomfort, and lowered the occurrence of severe VUR.