Hemifield-based analysis of pattern electroretinography in normal subjects and patients with preperimetric glaucoma.

This prospective cross-sectional study investigated the visual function of preperimetric glaucoma (PPG) patients based on hemifield (HF) pattern electroretinogram (PERG) amplitudes. Thirty-two (32) normal subjects and 33 PPG patients were enrolled in control and PPG groups, respectively. All of the participants had undergone full ophthalmic examinations, including spectral-domain optical coherence tomography (SD-OCT), visual field (VF) examination and pattern electroretinography (PERG). The PERG parameters along with the HF ratios of SD-OCT and PERG were compared between the control and PPG groups. Pairwise Pearson's correlation coefficients and linear regression models were fitted to investigate the correlations. The PERG N95 amplitudes were significantly lower in the PPG group (P < 0.001). The smaller/larger HF N95 amplitude ratio of the PPG group was found to be smaller than that of the control group (0.73 ± 0.20 vs. 0.86 ± 0.12; P = 0.003) and showed positive correlations with affected HF average ganglion cell-inner plexiform layer (GCIPL) thickness (r = 0.377, P = 0.034) and with average GCIPL thickness (r = 0.341, P = 0.005). The smaller/larger HF N95 amplitude ratio did not significantly change with age (ß = - 0.005, P = 0.195), whereas the full-field N95 amplitude showed a negative correlation with age (ß = - 0.081, P < 0.001). HF analysis of PERG N95 amplitudes might be particularly useful for patients with early glaucoma.

Orthotopic Mouse Models of Urinary Bladder Cancer.

BACKGROUND/AIM: Urinary bladder cancer has various etiologies and tends to recur and then progress to a higher grade. When muscles are invaded, the response to conventional therapy is poor and the quality of life deteriorates rapidly. Here, we summarize and compare two representative methods used to create the syngeneic mouse models required for immunological research. MATERIALS AND METHODS: In this study, we utilized six-week-old female C3H/HeNCrl mice and the mouse bladder tumor cell line MBT-2. The first method involved transurethral catheterization with poly-L-lysine pretreatment (catheter group), while the second method involved transperitoneal incision and direct injection of tumor cells into the bladder wall (open group). Mouse postoperative status was monitored on a weekly basis using magnetic resonance imaging (MRI). RESULTS: The catheter group had a tumor development rate of 47% (7 out of 15 mice), with only 1 mouse developing an intravesical tumor. In contrast, the open group had a higher tumor formation rate of 69% (47 out of 68 mice), with 27 mice showing intravesical tumor formation. Notably, with a lower cell count, urinary obstruction events were observed 2 weeks post-inoculation, which is one week later than the higher cell count group. CONCLUSION: In this study, we conducted a comparative analysis between the transurethral catheterization method and the transperitoneal incision and direct injection method in animal bladder tumor models. Our findings provide evidence of the consistent effectiveness in constructing a stable model within the open group. Well-designed orthotopic animal models are essential.

Analysis of stromal PDGFR-ß and α-SMA expression and their clinical relevance in brain metastases of breast cancer patients.

BACKGROUND: Breast cancer brain metastasis (BCBM) is a growing therapeutic challenge and clinical concern. Stromal cancer-associated fibroblasts (CAFs) are crucial factors in the modulation of tumorigeneses and metastases. Herein, we investigated the relationship between the expression of stromal CAF markers in metastatic sites, platelet-derived growth factor receptor-beta (PDGFR-ß), and alpha-smooth muscle actin (α-SMA) and the clinical and prognostic variables in BCBM patients. METHODS: Immunohistochemistry (IHC) of the stromal expression of PDGFR-ß and α-SMA was performed on 50 cases of surgically resected BCBM. The expression of the CAF markers was analyzed in the context of clinico-pathological characteristics. RESULTS: Expression of PDGFR-ß and α-SMA was lower in the triple-negative (TN) subtype than in other molecular subtypes (p = 0.073 and p = 0.016, respectively). And their expressions were related to a specific pattern of CAF distribution (PDGFR-ß, p = 0.009; α-SMA, p = 0.043) and BM solidity (p = 0.009 and p = 0.002, respectively). High PDGFR-ß expression was significantly related to longer recurrence-free survival (RFS) (p = 0.011). TN molecular subtype and PDGFR-ß expression were independent prognostic factors of recurrence-free survival (p = 0.029 and p = 0.030, respectively) and TN molecular subtype was an independent prognostic factor of overall survival (p < 0.001). CONCLUSIONS: Expression of PDGFR-ß in the stroma of BM was associated with RFS in BCBM patients, and the clinical implication was uniquely linked to the low expression of PDGFR-ß and α-SMA in the aggressive form of the TN subtype.

Different Expression and Clinical Implications of Cancer-Associated Fibroblast (CAF) Markers in Brain Metastases.

Aims: This study assessed the expression and clinical relevance of cancer-asssociated fibroblast (CAF)-related biomarkers in brain metastasis (BM). Moreover, molecular characterization of patient-derived primary CAFs and normal fibroblasts (NFs) was performed. Methods: Sixty-eight patients with BM from various primary cancer types were selected. Immunohistochemistry (IHC) and immunofluorescence (IF) staining were performed to evaluate the expression of various CAF-related biomarkers. CAFs and NFs were isolated from fresh tissues. Results: Various CAF-related biomarkers were expressed in CAFs in BMs of different primary cancers. However, only PDGFR-ß, α-SMA, and collagen type I were associated with BM size. PDGFR-ß and α-SMA were associated with BM recurrence after resection. PDGFR-ß was associated with recurrence-free survival (RFS). Interestingly, high expression of PDGFR-ß and α-SMA was found in the patients with previous chemotherapy or radiotherapy for primary cancer. In primary cell culture, PDGFR-ß and α-SMA were expressed at higher levels in patient-derived CAFs than in NFs or cancer cells. The origins of CAF in BM were presumed to be pericytes of blood vessels, circulating endothelial progenitor cells, or transformed astrocytes of the peritumoral glial stroma. Conclusion: Our results suggest that high expression of CAF-related biomarkers, particularly PDGFR-ß and α-SMA, is associated with poor prognosis and recurrence in patients with BM. With the elucidation of the role and origins of CAF in the tumor microenvironment, CAF can be a new imperative target for BM immunotherapy.

Refinement of recto-sigmoid colon vaginoplasty using a three-dimensional laparoscopic technique.

ABSTRACT: To investigate the feasibility, safety, and outcomes of three-dimensional (3D) laparoscopic vaginoplasty with a rectosigmoid colon flap for vaginal reconstruction.Following appropriate preoperative patient counseling, 17 consecutive patients underwent vaginoplasty using a 3D laparoscopic system. Perioperative and postoperative outcomes were retrospectively evaluated.Between September 2016 and February 2020, 17 patients underwent 3D laparoscopic vaginoplasty with a rectosigmoid colon flap. Of them, 15 (88%) were transgender female patients, and 2 (12%) were cisgender female patients with congenital deformities. Among the 15 transgender patients, 12 (80%) underwent de novo surgeries and 3 (20%) underwent re-do surgeries. The mean age at the time of operation was 33.0 years, and the mean total operation time was 529 ±â€Š128 minutes. The initial intraoperative mean vaginal depth was 15.2 ±â€Š1.3 cm, and the 30-day readmission rate was 5.9% (1/17 cases). The mean follow-up duration was 24.8 months.Perioperative and postoperative outcomes suggest that 3D laparoscopic rectosigmoid colon vaginoplasty is a potentially acceptable, effective, and safe method for vaginal reconstruction.

Impact of myopia on the association of long-term intraocular pressure fluctuation with the rate of progression in normal-tension glaucoma.

BACKGROUND/AIMS: To investigate whether the association of long-term intraocular pressure (IOP) fluctuation with the rate of progression of normal-tension glaucoma (NTG) differs between myopia and non-myopia. METHODS: The medical records of 65 myopic NTG (axial length (AL) > 24.0 mm) and 64 non-myopic NTG eyes (AL < 24.0 mm), who had been treated with topical medications for more than 5 years, were reviewed. Multiple linear regression models were fitted to analyse the relationships of the slope of mean deviation (MD) or visual field index (VFI) with the clinical factors, including the interactions with myopia. RESULTS: The average follow-up period was 8.3 years. Twenty-two (22) non-myopic eyes (34.4%) and 27 myopic eyes (41.5%) showed NTG progression (p=0.511). The interaction of myopia with IOP fluctuation was a significant factor regarding both MD and VFI slope (p=0.002, 0.024, respectively); stratified analyses suggested that the risk effect of IOP fluctuation was significant only in myopic NTG in terms of both MD (ß= -1.27, p=0.003) and VFI slope (ß=-2.32, p=0.011). CONCLUSION: Long-term IOP fluctuation was significantly related to faster visual field progression in myopic NTG eyes, compared with non-myopic NTG eyes.

A retrospective single-center study comparing clinical outcomes of 3-dimensional and 2-dimensional laparoscopic cholecystectomy in acute cholecystitis.

BACKGROUNDS/AIMS: Laparoscopic cholecystectomy (LC) has become widely used and preferred standard treatment for gallbladder (GB) disease in many countries. In this study, we aimed to compare the overall clinical outcomes of 3-dimensional (3D) LC system with those of the 2D LC method. METHODS: We retrospectively analyzed patients who underwent LC for acute cholecystitis between January 2010 and March 2019 at the National Medical Center in Korea. We entered them into 3D LC (group A) and 2D LC (group B) groups. We used Olympus CLV-190 laparoscopic device with dual lenses, capable of displaying both 3D and 2D images. Postoperative variables considered for evaluating between-group differences in clinical outcomes included diet resumption period after surgery, postoperative hospital length-of-stay, outpatient department follow-up period, surgical time, and postoperative surgery-related complications (blood loss and open conversion). RESULTS: We analyzed 278 acute cholecystitis patients (Group A, n=116; Group B, n=162). Compared to group B, group A had a significantly reduced surgical time and postoperative hospital stay. Although underlying diseases and abdominal surgical history were more prevalent in the 3D LC group, no significant between-group differences in blood loss and open conversion rate were observed. CONCLUSIONS: The 3D imaging system offered many advantages over 2D LC, including reduced surgical time and shorter postoperative hospital stay; therefore, it has significance in reducing hospital costs.

Caveolin-1 enhances brain metastasis of non-small cell lung cancer, potentially in association with the epithelial-mesenchymal transition marker SNAIL.

BACKGROUND: Caveolin-1 (Cav-1) plays an important role in the development of various human cancers. We investigated the relationship between Cav-1 expression and non-small cell lung cancer (NSCLC) progression in the context of brain metastasis (BM). METHODS: Cav-1 expression was investigated in a series of 102 BM samples and 49 paired primary NSCLC samples, as well as 162 unpaired primary NSCLC samples with (63 cases) or without (99 cases) metastasis to distant organs. Human lung cancer cell lines were used for in vitro functional analysis. RESULTS: High Cav-1 expression in tumor cells was observed in 52% (38/73) of squamous cell carcinomas (SQCs) and 33% (45/138) of non-SQCs. In SQC, high Cav-1 expression was increased after BM in both paired and unpaired samples of lung primary tumors and BM (53% vs. 84% in paired samples, P = 0.034; 52% vs. 78% in unpaired samples, P = 0.020). Although the difference in median overall survival in patients NSCLC was not statistically significant, high Cav-1 expression in tumor cells (P = 0.005, hazard ratio 1.715, 95% confidence index 1.175-2.502) was independent prognostic factors of overall survival on multivariate Cox regression analyses, in addition to the presence of BM and non-SQC type. In vitro assays revealed that Cav-1 knockdown inhibited the invasion and migration of lung cancer cells. Genetic modulation of Cav-1 was consistently associated with SNAIL up- and down-regulation. These findings were supported by increased SNAIL and Cav-1 expression in BM samples of SQC. CONCLUSIONS: Cav-1 plays an important role in the BM of NSCLC, especially in SQC. The mechanism may be linked to SNAIL regulation.

The Role Played by SLUG, an Epithelial-Mesenchymal Transition Factor, in Invasion and Therapeutic Resistance of Malignant Glioma.

In malignant gliomas, invasive phenotype and cancer stemness promoting resurgence of residual tumor cells render treatment very difficult. Hence, identification of epithelial-mesenchymal transition (EMT) factors associated with invasion and stemness of glioma cells is critical. To address the issue, we investigated several EMT factors in hypermotile U87MG and U251 cells, orthotopic mouse glioma model, and human glioma samples. Of several EMT markers, SLUG expression was notably increased at the invasive fronts of gliomas, both in mouse tumor grafts and human glioma samples. The biological role played by SLUG was investigated using a colony-forming assay after chemotherapy and irradiation, and by employing a neurosphere culture assay. The effect of SLUG on glioma progression was examined in our patient cohort and samples, and compared to large public data from the REMBRANDT and TCGA. Genetic upregulation of SLUG was associated with increased levels of stemness factors and enhanced resistance to radiation and temozolomide. In our cohort, patients exhibiting lower-level SLUG expression evidenced longer progression-free survival (P = 0.042). Also, in the REMBRANDT dataset, a group in which SLUG was downregulated exhibited a significant survival benefit (P < 0.001). Although paired glioblastoma samples from our patients did not show a significant increase of SLUG expression, increased mRNA levels of SLUG were found in recurrent glioblastoma from TCGA (P = 0.052), and in temozolomide-treated glioma cells and mouse tumor grafts. SLUG may contribute to glioma progression by controlling invasion at infiltrating margins, associated with increased stemness and therapeutic resistance.

Prognostic significance of E-cadherin and N-cadherin expression in Gliomas.

BACKGROUND: Epithelial-mesenchymal transition (EMT), principally involving an E-cadherin to N-cadherin shift, linked to tumor invasion or metastasis, and therapeutic resistance in various human cancer. A growing body of recent evidence has supported the hypothesis that EMT play a crucial role in the invasive phenotype of gliomas. To evaluate the prognostic connotation of EMT traits in glioma, expression of E-cadherin and N-cadherin was explored in a large series of glioma patients in relation to patient survival rate. METHODS: Expressions of E- and N-cadherin were examined using immunohistochemical analysis in 92 glioma cases diagnosed at our hospital. These markers expressions were also explored in 21 cases of fresh frozen glioma samples and in glioma cell lines by Western blot analysis. RESULTS: Expression of E-cadherin was observed in eight cases (8.7%) with weak staining intensity in the majority of the immunoreactive cases (7/8). Expression of N-cadherin was identified in 81 cases (88.0%) with high expression in 64 cases (69.5%). Fresh frozen tissue samples and glioma cell lines showed similar results by Western blot analysis. There was no significant difference in either overall survival (OS) or progression-free survival (PFS) according to E-cadherin expression (P > 0.05). Although the OS rates were not affected by N-cadherin expression levels (P = 0.138), PFS increased in the low N-cadherin expression group with marginal significance (P = 0.058). The survival gains based on N-cadherin expression levels were significantly augmented in a larger series of publicly available REMBRANDT data (P < 0.001). CONCLUSIONS: E- and N-cadherin, as representative EMT markers, have limited prognostic value in glioma. Nonetheless, the EMT process in gliomas may be compounded by enhanced N-cadherin expression supported by unfavorable prognostic outcomes.

Human U87 glioblastoma cells with stemness features display enhanced sensitivity to natural killer cell cytotoxicity through altered expression of NKG2D ligand.

BACKGROUND: Glioblastoma (GBM) is one of the most lethal tumors with a poor prognosis. Its inevitable recurrence is frequently explained by the presence of cancer stem cells. We aimed to show that human GBM cells with stemness features are more sensitive to natural killer (NK) cells than GBM cells without stemness characteristics. METHODS: Natural killer cell cytotoxicity was measured using flow cytometry in neurosphere-forming U87 GBM cells cultured with neurobasal media (NBE condition) and compared with that in serum-cultured U87 GBM cells (serum condition). Cytotoxicity was examined after addition of blocking NKG2D monoclonal antibodies. The expression profile of NK ligands of NK cells were investigated by reverse transcription polymerase chain reaction and western blot analysis in the U87 GBM cells in both conditions. RESULTS: NBE U87 cells showed higher cytotoxicity to NK cells than serum U87 cells did (55 vs 35% at an effector to target cell ratio of 5:1). The increased cytotoxicity was diminished in NBE U87 cells by a larger gap than in serum U87 cells by adding NKG2D blocking antibodies. Of the NKG2D ligands, the expression of ULBP1 and ULBP3 was relatively increased in NBE U87 cells compared to serum U87 cells. CONCLUSIONS: U87 GBM cells with stemness features demonstrate increased cytotoxicity to NK cells in association with altered NKG2D ligand expression of NK cell activating receptor. Applying immune modulation to GBM treatment may be a promising adjuvant therapy in patients with intractable GBM.

KITENIN promotes glioma invasiveness and progression, associated with the induction of EMT and stemness markers.

KITENIN (KAI1 COOH-terminal interacting tetraspanin) promotes tumor invasion and metastasis in various cancers. This study assessed the association between KITENIN expression and advanced glioma grade in patients. In vitro assays revealed that KITENIN knockdown inhibited the invasion and migration of glioma cells, whereas KITENIN overexpression promoted their invasion and migration. In orthotopic mouse tumor models, mice transplanted with KITENIN-transfected glioma cells had significantly shorter survival than mice transplanted with mock-transfected cells. Patients with low KITENIN expression showed a significantly longer progression-free survival than patients with high KITENIN expression. KITENIN induced the expression of the epithelial-mesenchymal transition (EMT) markers (N-cadherin, ZEB1, ZEB2, SNAIL and SLUG) as well as the glioma stemness markers (CD133, ALDH1 and EPH-B1). Taken together, these findings showed that high levels of KITENIN increased glioma invasiveness and progression, associated with the up-regulation of EMT and stemness markers.

Association of immune status with recurrent anal condylomata in human immunodeficiency virus-positive patients.

PURPOSE: An anal condyloma is a proliferative disease of the genital epithelium caused by the human papillomavirus. This condition is most commonly seen in male homosexuals and is frequently recurrent. Some reports have suggested that immunosuppression is a risk factor for recurrence of a condyloma. Thus, we investigated the risk factors for a recurrent anal condyloma in human immunodeficiency virus (HIV)-positive patients. METHODS: We retrospectively analyzed 85 consecutive patients who were diagnosed with and underwent surgery for an anal condyloma from January 2007 to December 2011. Outcomes were analyzed based clinical and immunologic data. RESULTS: Recurrent anal condylomata were found in 25 patients (29.4%). Ten cases (40.0%) were within postoperative 3 months. At postoperative 6 months, the CD4 lymphocyte count in the recurrent group was lower than it was in the nonrecurrent group (P = 0.023). CONCLUSION: CD4-mediated immunosuppression is a risk factor for recurrent anal condylomata in HIV-positive patients.

A Clinical Comparison of Laparoscopic versus Open Appendectomy for Complicated Appendicitis.

PURPOSE: Although laparoscopic appendectomies (LAs) are performed universally, a controversy still exists whether the LA is an appropriate surgical approach to complicated appendicitis (CA). We retrospectively evaluated the outcomes of laparoscopic versus open appendectomies for CA. METHODS: We retrospectively analyzed 60 consecutive patients who were diagnosed as having CA from July 2009 to January 2011. Outcomes such as operative time, time to soft diet, length of hospital stay, and postoperative complications were analyzed. RESULTS: There were no statistically significant differences in operative time between the LA and the open appendectomy (OA) groups. Return to soft diet was faster in the LA group (2.1 ± 1.2 vs. 3.5 ± 1.5 days; P = 0.001). Length of hospital stay was shorter for the LA group (4.4 ± 2.3 vs. 5.8 ± 2.9 days; P = 0.045). The overall complication rates showed no statistically significant difference between the two groups. In cases involving a periappendiceal abscess, the LA had a significantly higher incidence of intra-abdominal abscess (IAA) and postoperative ileus (PI; P = 0.028). CONCLUSION: The LA showed good results in terms of the time to soft diet, the length of hospital stay, and surgical site infection (SSI) whereas the overall complication rates were similar for the two groups. However, the LA was associated with significantly higher incidence of IAA and PI for the cases with a periappendiceal abscess. Therefore, when using a LA, the surgeon must take great care to minimize the incidence of IAA and PI if a periappendiceal abscess is present.

The Early Experience of Laparoscopy-assisted Gastrectomy for Gastric Cancer at a Low-volume Center.

PURPOSE: Laparoscopy-assisted gastrectomy (LAG) has become a technically feasible and safe procedure for early gastric cancer treatment. LAG is being increasingly performed in many centers; however, there have been few reports regarding LAG at low-volume centers. The aim of this study was to report our early experience with LAG in patients with gastric cancer at a low-volume center. MATERIALS AND METHODS: The clinicopathologic data and surgical outcomes of 39 patients who underwent LAG for gastric cancer between April 2007 and March 2010 were retrospectively reviewed. RESULTS: The mean age was 68.3 years. Thirty-one patients had medical co-morbidities. The mean patient ASA score was 2.0. Among the 39 patients, 4 patients underwent total gastrectomies and 35 patients underwent distal gastrectomies. The mean blood loss was 145.4 ml and the mean operative time was 259.4 minutes. The mean time-to-first flatus, first oral intake, and the postoperative hospital stay was 2.8, 3.1, and 9.3 days, respectively. The 30-day mortality rate was 0%. Postoperative complications developed in 9 patients, as follows: anastomotic leakage, 1; wound infection, 1; gastric stasis, 2; postoperative ileus, 1; pneumonia, 1; cerebral infarction, 1; chronic renal failure, 1; and postoperative psychosis, 1. CONCLUSIONS: LAG is technically feasible and can be performed safely at a low-volume center, but an experienced surgical team and careful patient selection are necessary. Furthermore, for early mastery of the learning curve for LAG, surgeons need education and training in addition to an accumulation of cases.