Bee venom inhibits the proliferation and migration of cervical-cancer cells in an HPV E6/E7-dependent manner.

Bee venom (BV), secreted from the venom gland of the honey bee, contains several biological active compounds. BV has been widely used as a traditional medicine for treating human disease, including cancer. In this study, we have shown the molecular mechanism underlying the therapeutic effect of BV on cancer. Treatment with BV reduced the proliferation of cervical-cancer cells in a dose- and time-dependent manner. Interestingly, the killing effect of BV was specific to HPVpositive cervical-cancer cell lines, such as Caski and HeLa cells, and not to HPV-negative cervical-cancer cells (C33A). BV reduced the expression of HPV E6 and E7 at RNA and protein levels, leading to an increase in the expression of p53 and Rb in Caski and HeLa cells. Further, BV decreased the levels of cell-cycle proteins, such as cyclin A and B, and increased the levels of cell-cycle inhibitors, such as p21 and p27. BV significantly induced apoptosis and inhibited wound healing and migration of cervical-cancer cells. It also upregulated the expression of pro-apoptotic BAX and downregulated the expression of anti-apoptotic Bcl-2 and Bcl-XL. Cleavage of caspase-3, caspase-9, and PARP were also induced by BV treatment, whereas the phosphorylation of mitogenic signalingrelated proteins, such as AKT, JNK, p38, and ERK, were downregulated. Our results indicate that BV has a therapeutic selectivity for HPV-positive malignant cells, so further clinical studies are needed to assess its clinical application. [BMB Reports 2020; 53(8): 419-424].

Predictors of biochemical recurrence after Retzius-sparing robot-assisted radical prostatectomy: Analysis of 359 cases with a median follow-up period of 26 months.

OBJECTIVES: Our aim was to evaluate the predictors of biochemical recurrence after Retzius-sparing robot-assisted radical prostatectomy. METHODS: The study cohort consisted of 359 consecutive non-metastatic prostate cancer patients who underwent Retzius-sparing robot-assisted radical prostatectomy between November 2012 and January 2016. According to the National Comprehensive Cancer Network prostate cancer risk classification, 164 patients (45.7%) had high- or very high-risk prostate cancer. No patient received adjuvant therapy until documented biochemical recurrence. Biochemical recurrence-free survival was estimated using the Kaplan-Meier method. Univariable and multivariable Cox proportional hazards regression models were used to determine variables predictive of biochemical recurrence. RESULTS: The median follow-up period was 26 months (interquartile range 19-38 months). The overall biochemical recurrence rate was 14.8%, and the median time to biochemical recurrence was 11 months (interquartile range 6-22 months). The 3-year biochemical recurrence-free survival probability was 71.2%, 72.1%, 88.7%, 82.3% and 95.7% in very high-, high-, intermediate-, low- and very low-risk prostate cancer, respectively (log-rank, P < 0.001). On multivariable analysis, preoperative prostate-specific antigen (hazard ratio 1.03, 95% confidence interval 1.02-1.04; P < 0.0001), percentage of maximum core involvement on biopsy (hazard ratio 1.02, 95% confidence interval 1.01-1.03; P = 0.029) and clinical stage ≥T3a (hazard ratio 2.12, 95% confidence interval 1.02-4.39; P = 0.043) were predictors of biochemical recurrence, whereas pathological Gleason score ≥8 (hazard ratio 5.63, 95% confidence interval 1.62-19.61; P = 0.007) and pathological tumor volume (hazard ratio 1.08, 95% confidence interval 1.04-1.20; P < 0.001) were the main pathological predictors of biochemical recurrence. CONCLUSIONS: Retzius-sparing robot-assisted radical prostatectomy confers effective biochemical recurrence control at the mid-term follow-up period. Preoperative prostate-specific antigen, advanced clinical stage and higher Gleason score were important predictors of biochemical recurrence after Retzius-sparing robot-assisted radical prostatectomy. Long-term oncological safety still needs to be established.

Changes in the optic nerve head induced by horizontal eye movements.

PURPOSE: To investigate the effect of eye movement on the optic nerve head (ONH) using swept-source optical coherence tomography (SS-OCT), and to measure the degree of ONH changes. METHODS: We enrolled 52 healthy subjects, 20 to 40 years of age, and performed a prospective observational study. Both ONH and macula were imaged simultaneously using wide volume scan of the SS-OCT in the primary and different gaze positions. Horizontal eye movements were used to obtain OCT images in abducted and adducted eyeball positions. Multilateral 3-dimensional registration was used to process and analyze the images to measure the degree of ONH changes. RESULTS: The mean axial length (AXL) was 25.73 ± 1.42mm and the mean spherical equivalents was -4.49 ± 2.94 D (The proportion of high myopia was 39.4%). Significant morphologic changes were observed in the ONH during both abduction and adduction. In abduction, the overall ONH tissues were elevated, and the mean area of elevation was 115,134 ± 9,424 µm2 (p<0.001). In adduction, the mean areas from two perspectives, which were nasal or temporal, and peripapillary tissues or optic nerve cupping were 95,277 ± 73,846 µm2, 34,450 ± 44,948 µm2, -108,652 ± 91,246 µm2, and -30,581 ± 46,249 µm2, respectively. Elevation in abduction (overall, nasal cup segment, and temporal cup segment; R = 0.204, 0.195 and 0.225, p = 0.038, 0.047 and 0.021, respectively) and elevation of nasal peripapillary segments in adduction were positively correlated with AXL (R = 0.346, p<0.001). CONCLUSION: We found significant morphologic changes in the ONH in both abduction and adduction and these changes were associated with AXL. Considering these morphologic changes as physical properties, it allows a better understanding of the biomechanical characteristics of the ONH.

Extracellular superoxide dismutase ameliorates house dust mite-induced atopic dermatitis-like skin inflammation and inhibits mast cell activation in mice.

Extracellular superoxide dismutase (EC-SOD) is an enzyme that catalyses the dismutation of superoxide anions. It has multiple functions, such as reactive oxygen species scavenging, anti-angiogenic, anti-inflammatory, antichemotatic and antitumor activities. Recently, we demonstrated that EC-SOD inhibits ovalbumin-induced allergic airway inflammation in mice. However, the anti-allergic effect of EC-SOD on skin tissue and the role of EC-SOD in mast cells, which are important for allergic responses, have not been well studied. In this study, we investigated whether EC-SOD can alleviate atopic dermatitis in mice and inhibit mast cell activation. Treatment with human recombinant EC-SOD ameliorated house dust mite-induced atopic dermatitis in mice. Furthermore, the levels of pro-allergic cytokine gene expression and histamine release increased in EC-SOD KO mast cells and decreased in EC-SOD overexpressing mast cells, suggesting that EC-SOD inhibits mast cell activation. Consistently, a passive cutaneous anaphylaxis experiment showed more blood leakage from EC-SOD KO mouse ear skin, implying that the lack of EC-SOD increases allergic responses. These results suggest that EC-SOD inhibits mast cell activation and atopic dermatitis and that the loss of EC-SOD causes more severe allergic responses, implying that EC-SOD might be a good drug candidate for treatment of allergic disorders, such as atopic dermatitis.

Transscleral Fixation of Intraocular Lenses Using Modified Injector

PURPOSE: To evaluate the clinical stability and outcomes of 3-piece intraocular lens (IOL) transscleral fixation surgery using a modified injector. METHODS: We have modified and used the Sapphire unfolder injector system (Allergan(R), USA). This involved, cutting a slit longitudinally at the terminal part of the injector so that a thread could pass through it freely. After a conjunctival peritomy created at 2 and 8 o'clock, a long curved needle with double-armed 10-0 polypropylene is passed through the exposed sclera. Two pieces of suture are withdrawn through the 2.8 mm corneal incision and 1 suture (from 8 o'clock) is passed through the opening of the cartridge and then tied to the leading haptic. Next, the IOL was implanted with the cartridge and then inserted through the corneal incision site. The other suture (from 2 o'clock) is tied to the haptic on the opposite side and inserted. RESULTS: The study included 20 eyes of 20 patients with a mean age of 62.8 years at the initial visit. There were no complications, such as vitreous hemorrhage, retinal detachment, glaucoma, corneal edema, or iris injury. While the knot fixed to the leading haptic of IOL passed by the cartridge, there was no change of position. During the follow-up period, IOL dislocation did not occur and the corrected visual acuity and corneal astigmatism improved significantly. CONCLUSIONS: This technique is an effective procedure for minimizing entangled thread and corneal astigmatism.

Synchronous activation of gonadotropin-releasing hormone gene transcription and secretion by pulsatile kisspeptin stimulation.

Pulsatile release of hypothalamic gonadotropin-releasing hormone (GnRH) is essential for pituitary gonadotrope function. Although the importance of pulsatile GnRH secretion has been recognized for several decades, the mechanisms underlying GnRH pulse generation in hypothalamic neural networks remain elusive. Here, we demonstrate the ultradian rhythm of GnRH gene transcription in single GnRH neurons using cultured hypothalamic slices prepared from transgenic mice expressing a GnRH promoter-driven destabilized luciferase reporter. Although GnRH promoter activity in each GnRH neuron exhibited an ultradian pattern of oscillations with a period of ∼10 h, GnRH neuronal cultures exhibited partially synchronized bursts of GnRH transcriptional activity at ∼2-h intervals. Surprisingly, pulsatile administration of kisspeptin, a potent GnRH secretagogue, evoked dramatic synchronous activation of GnRH gene transcription with robust stimulation of pulsatile GnRH secretion. We also addressed the issue of hierarchical interaction between the circadian and ultradian rhythms by using Bmal1-deficient mice with defective circadian clocks. The circadian molecular oscillator barely affected basal ultradian oscillation of GnRH transcription but was heavily involved in kisspeptin-evoked responses of GnRH neurons. In conclusion, we have clearly shown synchronous bursts of GnRH gene transcription in the hypothalamic GnRH neuronal population in association with episodic neurohormone secretion, thereby providing insight into GnRH pulse generation.

Empirical use of ciprofloxacin for acute uncomplicated pyelonephritis caused by Escherichia coli in communities where the prevalence of fluoroquinolone resistance is high.

There is little information about the effectiveness of ciprofloxacin in regions where ciprofloxacin-resistant Escherichia coli is prevalent. This study was conducted to evaluate whether ciprofloxacin is effective as the initial empirical antibiotic for treatment of uncomplicated acute pyelonephritis (APN) due to ciprofloxacin-resistant E. coli. A total of 255 women with clinical diagnoses of uncomplicated APN due to E. coli were enrolled in the emergency department between March 2005 and December 2008. All enrolled patients were initially treated with ciprofloxacin. Patients were followed up 4 to 7 days after the start of therapy and 14 to 21 days after its completion. At the first follow-up visit, ciprofloxacin was changed to the appropriate antibiotic when necessary, depending on the antibiotic susceptibility results. Not only improvement of symptoms and signs but also microbiologic eradication was assessed at each visit. Fifteen percent (39/255) of the E. coli isolates were resistant to ciprofloxacin. There was no statistically significant difference between the clinical cure rates of the ciprofloxacin-susceptible group and the ciprofloxacin-resistant group at the first follow-up (87.0% versus 76.9%, P = 0.135) or the second follow-up (98.6% versus 94.9%, P = 0.177). However, there was a lower microbiologic cure rate in the ciprofloxacin-resistant group than in the ciprofloxacin-susceptible group (92.4% versus 41.7%, P = 0.000) at the first follow-up visit. No complications occurred in the ciprofloxacin-resistant group during the follow-up period. Our findings indicate that ciprofloxacin is an appropriate choice for empirical therapy of uncomplicated APN and has no serious adverse outcomes, if it is tailored appropriately, even for women infected with ciprofloxacin-resistant E. coli.

Protective effect of the methanolic extract from Duchesnea indica against oxidative stress in vitro and in vivo.

Duchesnea indica (Rosaceae family) is herb used extensively in traditional Chinese medicine. In this study we investigated its protective activity against hydrogen peroxide (H(2)O(2))-induced cytotoxicity in human skin fibroblast (CCD-986Sk) cells and 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced H(2)O(2) in the skin of hairless mice. Pretreatment of CCD-986Sk cells with methanolic extract of D. indica (DIM) improved the cell viability, enhanced activity of catalase, and decreased the leakage of lactate dehydrogenase (LDH) and the levels of malondialdehyde (MDA) and intracellular reactive oxygen species (ROS) in H(2)O(2) injured cells. Furthermore, DIM inhibited cell apoptosis and Bax expression induced by H(2)O(2). In addition, the level of H(2)O(2) stimulated by TPA was decreased by DIM in the skin of hairless mice. These results suggest that DIM offers protection against oxidative stress in vitro and in vivo, and this ability suggests potential use for protection against oxidation-induced skin damage.

The effects of chronic treatment with Morus bombycis KOIDZUMI in spontaneously hypertensive rats.

The present study was performed to evaluate the antihypertensive effects of Morus bombycis KOIDZUMI (MK) in spontaneously hypertensive rats (SHRs). In addition, the effects on vascular responses and cardiac functions were also investigated. In isolated rat aortic preparations, the 100% ethanol extract of MK exhibited a potent vascular relaxant effect with IC(50) value of 3.9 microg/ml, and this vasorelaxant effect was completely abolished by pretreatment of the aortic tissues with N(G)-nitro-L-arginine methyl ester or the denudation of endothelial layer. In isolated rat hearts, the MK extract significantly reduced cardiac functions such as left ventricular developed pressure and heart rate. In an antihypertensive study in SHRs, long-term administration with MK extracts (10, 30, 100 mg/kg) for 42 d dose-dependently decreased systolic blood pressure (approximately 20 mmHg). In SHRs, MK extract enhanced the aortic relaxation to acetylcholine and sodium nitroprusside after 42 d of treatment. In addition, lipid peroxidation and DNA damage in liver of SHRs were also attenuated by long-term treatment with MK extract. These results suggest that chronic treatment with MK extract exerts an antihypertensive effect in SHRs, and its direct vasorelaxant, negative inotropic actions, and anti-oxidant properties may contribute to reduce the elevated blood pressure.

Identification and properties of 2,5-dihydroxy-4,3'-di(beta-D-glucopyranosyloxy)-trans-stilbene from Morus bombycis Koidzumi roots.

2,5-Dihydroxy-4,3'-di(beta-d-glucopyranosyloxy)-trans-stilbene was identified from Morus bombycis Koidzumi roots. The 2,5-dihydroxy-4,3'-di(beta-D-glucopyranosyloxy)-trans-stilbene at a dose of 400-600 mg/kg had hepatoprotective activity comparable to the standard agent, silymarin. The biochemical assays were confirmed by histological observations showing that the 2,5-dihydroxy-4,3'-di(beta-d-glucopyranosyloxy)-trans-stilbene from Morus bombycis Koidzumi roots decreased cell ballooning in response to CCl4 treatment. These results demonstrate that the 2,5-dihydroxy-4,3'-di(beta-d-glucopyranosyloxy)-trans-stilbene component has a liver protective action against CCl4-induced hepatotoxicity.

Antioxidant effects and hepatoprotective activity of 2,5-dihydroxy-4,3'-di(beta-d-glucopyranosyloxy)-trans-stilbene from Morus bombycis Koidzumi roots on CCl4-induced liver damage.

We investigated hepatoprotective activity and antioxidant effect of the 2,5-dihydroxy-4,3'-di(beta-D-glucopyranosyloxy)-trans-stilbene that purified from Morus bombycis Koidzumi roots against CCl4-induced liver damage in rats. The 2,5-dihydroxy-4,3'-di(beta-D-glucopyranosyloxy)-trans-stilbene displayed dose-dependent superoxide radical scavenging activity (IC50 = 430.2 microg/ml), as assayed by the electron spin resonance (ESR) spin-trapping technique. The increase in aspartate aminotransferase (AST) activities in serum associated with carbon tetrachloride (CCl4)-induced liver injury was inhibited by 2,5-dihydroxy-4,3'-di(beta-D-glucopyranosyloxy)-trans-stilbene and at a dose of 400 - 600 mg/kg samples had hepatoprotective activity comparable to the standard agent, silymarin. The biochemical assays were confirmed by histological observations showing that the 2,5-dihydroxy-4,3'-di(beta-d-glucopyranosyloxy)-trans-stilbene decreased cell ballooning in response to CCl4 treatment. These results demonstrate that the 2,5-dihydroxy-4,3'-di(beta-D-glucopyranosyloxy)-trans-stilbene is a potent antioxidant with a liver protective action against CCl4-induced hepatotoxicity.

A Clinicostatistical Study of Congenital Alimentary Tract Obstruction

Clinicostatistical datsa on 314 patients with congenital alimentary tract obstruction at the Department of Pediatrics and Pediatric Surgery of Seoul National University Hospital from January, 1974 to July, 1980 are presented and reviewed. The results are as follows; 1. Congenital megacolon(133 cases) is the most common congenital alimentary tract obstruction, and anorectal malformations(127 cases), congental duodenal obstruction(20 cass), esophageal atresia(12 cases), ileal atresia(9 cases), multiple intestinal atresia(2 cases), and colon atresia(2 cases), in given order of frequency. 2. The 20 cases of congenital duodenal obstruction included 15 male nad 5 female patients. The 10 cases were intrinsic obstruction(diaphragm of web; 4, atresia; 3, stenosis; 2, uncertain; 1) and the 10 cases were extrinsic obstruction(malrotation with or without Ladd's band, midgut volvulus: 9, preduodenal portal vein; 1). Associated anomalies were present in 3(1%) of the total. Operation was carried out in 19 patients and seven of them died. 3. The 22 cases of ileo-jejuno-colon atresia and stenosis included 14 male and 4 female patients. The majority of the patients involved ileum(9 cases), jejunum(9 cases), Jejunoileum(2 cases), and colon aresia was present in 2 cases. Associate anomalies were present in 7(32%) of the total. Operation was performed in 20 patients and seven of them died. 4. The 133 cases of congenital megacolon included 99 male and 34 female patients. The 121 case(91%) were short segment involvement and the remainders were long segment involovement. The 10 cases(8%) were associated with other malformations. In 118 patients operation was performed and the remainders were received conservative treatments, including enema. The 20 cases, 18% of the patients developed complications such as enterocolitis, incontinence, and so on. Three patients died during follow-up.

A Clinicostatistical Study of Congenital Alimentary Tract Obstruction

Clinicostatistical datsa on 314 patients with congenital alimentary tract obstruction at the Department of Pediatrics and Pediatric Surgery of Seoul National University Hospital from January, 1974 to July, 1980 are presented and reviewed. The results are as follows; 1. Congenital megacolon(133 cases) is the most common congenital alimentary tract obstruction, and anorectal malformations(127 cases), congental duodenal obstruction(20 cass), esophageal atresia(12 cases), ileal atresia(9 cases), multiple intestinal atresia(2 cases), and colon atresia(2 cases), in given order of frequency. 2. The 20 cases of congenital duodenal obstruction included 15 male nad 5 female patients. The 10 cases were intrinsic obstruction(diaphragm of web; 4, atresia; 3, stenosis; 2, uncertain; 1) and the 10 cases were extrinsic obstruction(malrotation with or without Ladd's band, midgut volvulus: 9, preduodenal portal vein; 1). Associated anomalies were present in 3(1%) of the total. Operation was carried out in 19 patients and seven of them died. 3. The 22 cases of ileo-jejuno-colon atresia and stenosis included 14 male and 4 female patients. The majority of the patients involved ileum(9 cases), jejunum(9 cases), Jejunoileum(2 cases), and colon aresia was present in 2 cases. Associate anomalies were present in 7(32%) of the total. Operation was performed in 20 patients and seven of them died. 4. The 133 cases of congenital megacolon included 99 male and 34 female patients. The 121 case(91%) were short segment involvement and the remainders were long segment involovement. The 10 cases(8%) were associated with other malformations. In 118 patients operation was performed and the remainders were received conservative treatments, including enema. The 20 cases, 18% of the patients developed complications such as enterocolitis, incontinence, and so on. Three patients died during follow-up.