PURPOSE: To investigate the effect of estrogen on the expression of calcium-activated potassium (KCa) channels in an overactive bladder rat model. To this end, mRNA and protein levels of KCa channel subtypes in the bladder of ovariectomized rats were measured by reverse transcription polymerase chain reaction and western blotting, respectively. METHODS: Ten-week-old female Sprague-Dawley rats were divided randomly into 3 groups: sham-operated control group (n=11), ovariectomy group (n=11), and the group treated with estrogen after ovariectomy (n=12). Rats in the last group were subcutaneously injected with 17β-estradiol (50 µg/kg) every other day for 2 weeks, whereas rats in the other 2 groups received vehicle (soybean oil) alone. Two weeks after treatment, the whole bladder was excised for mRNA and protein measurements. RESULTS: Protein levels of the large-conductance KCa (BK) channels in the ovariectomy group were 1.5 folds higher than those in the sham-operated control group. However, the protein levels of the other KCa channel subtypes did not change significantly upon bilateral ovariectomy. Treatment with 17β-estradiol after ovariectomy restored BK channel protein levels to the control value. In contrast, BK channel mRNA levels were not significantly affected by either ovariectomy alone or 17β-estradiol treatment. The small-conductance KCa type 3 channel (SK3) mRNA and protein levels decreased to 75% of control levels upon 17β-estradiol treatment. CONCLUSIONS: These results suggest that 17β-estradiol may influence urinary bladder function by modulating BK and SK3 channel expression.
Animals ; Blotting, Western ; Estrogens ; Female ; Humans ; Models, Animal ; Ovariectomy ; Polymerase Chain Reaction ; Potassium ; Potassium Channels, Calcium-Activated* ; Rats* ; Rats, Sprague-Dawley ; Reverse Transcription ; RNA, Messenger ; Urinary Bladder* ; Urinary Bladder, Overactive

PURPOSE: Recent studies have shown that potassium (K+) and sodium channels are involved in prostate cell growth. However, a great many of the studies have been done in prostate cancer cell lines and there are only scant studies on prostate cancer and benign prostatic hypertrophy (BPH) tissue. The present study was aimed to evaluate the alterations of the calcium-activated K+ channel (KCa) expression in prostate cancer, and to compare them with the expression profiles in human BPH tissue to understand their potential role in the progression of prostate cancer. MATERIALS AND METHODS: The prostate tissues obtained from radical prostatectomy (n=10) and transurethral resection of the prostate (n=18) were quickly frozen in liquid nitrogen for the RNA measurements. The protein and mRNA levels of the KCa subtypes and connexins were measured by performing immunoblot analysis and reverse-transcription polymerase chain reaction, respectively. RESULTS: The mRNA levels of type 2 (SK2) and type 3 (SK3) small-conductance and large-conductance (BK) KCas in the prostate cancer tissues were decreased more than 50% compared with those in the BPH samples. In addition, the BK and SK2 protein levels in prostate cancer were also significantly lower than those in the BPH. As reported previously, the connexin 26 and 43 transcript signals in the prostate cancer were significantly reduced compared with those in the BPH samples. CONCLUSIONS: These results suggest that the impaired expression of KCas may have a role in tumor progression via aberrant and uncontrolled prostate cell growth.
Cell Line ; Connexins ; Humans* ; Large-Conductance Calcium-Activated Potassium Channels ; Nitrogen ; Polymerase Chain Reaction ; Potassium ; Potassium Channels, Calcium-Activated* ; Prostate* ; Prostatectomy ; Prostatic Hyperplasia ; Prostatic Neoplasms* ; RNA ; RNA, Messenger ; Small-Conductance Calcium-Activated Potassium Channels ; Sodium Channels

PURPOSE: The aim of this study was to examine how the mRNA and protein levels of calcium activated Kchannel (K(Ca)) and connexin (Cx) change in association with overactive bladder in the bladder mucosae of stress urinary incontinence (SUI) patients. MATERIALS AND METHODS: Twenty SUI patients were included in our study. Bladder mucosae were obtained, with using cold cup biopsy forceps, from the patients suffering with genuine stress urinary incontinence (group 1, n=7), from the patients suffering with SUI along with urgency and frequency (group 2, n=6), and from the patients suffering with mixed incontinence (group 3, n=7). RESULTS: The mRNA transcripts of type 2 (SK2) and type 3 (SK3) small conductance K(Ca), Cx26, and Cx43 were highly expressed in the bladder mucosa. The message of large conductance K(Ca)(BK) was significantly decreased in group 3 compared with that in the controls. The SK2 and Cx26 messages in group 3 were also lower than those in groups 1 and 2. In the presence of urge incontinence, the BK and SK2 protein levels were decreased and the Cx26 protein expression was significantly increased in the bladder mucosa of the SUI patients. In contrast, there were no significant differences in the mRNA and protein levels of K(Ca)s and Cxs between groups 1 and group 2. CONCLUSIONS: Downregulation of both BK and SK2 and upregulation of Cx26 in the bladder mucosa of MI patients may contribute to the alterations of urothelial instability, and this correlate with the symptom severity of bladder instability in SUI patients.
Biopsy ; Calcium ; Connexin 43 ; Connexins ; Down-Regulation ; Humans ; Mucous Membrane* ; Potassium Channels, Calcium-Activated* ; RNA, Messenger ; Surgical Instruments ; Up-Regulation ; Urinary Bladder* ; Urinary Bladder, Overactive* ; Urinary Incontinence* ; Urinary Incontinence, Stress ; Urinary Incontinence, Urge ; Urothelium

Purpose: Recent studies have shown that transient receptor potential melastatin-subfamily member 7 (TRPM7) may enhance cancer cell growth, migration and invasion of human renal cell carcinoma (RCC). We investigated how TRPM7 regulated progression of RCC by interacting with matrix metalloproteinase (MMP)/tissue inhibitors of metalloproteinase (TIMP) pathway.
Materials and Methods: We performed a wound healing assay and a transwell migration to examine the migration of RCC cells and transwell invasion assay to assess the invasion of RCC cells. Western blot analysis was used to show the expression of MMP-2, MMP-9, TIMP-1, and TIMP-2.
Results: The migration and invasion of RCC cells were markedly suppressed by siRNA targeting TRPM7. Lowering of TRPM7 increased MMP-2 expression and induced no change in MMP-9 expression. Strikingly, TRPM7 silencing suppressed the expression of TIMP-1 and TIMP-2.
Conclusions These results suggested that MMP-independent action of TIMPs may take part in the enhancing effect of TRPM7 on the progression of RCC. (Korean J Urol Oncol 2020;18:61-67)

PURPOSE: To evaluate the therapeutic potential of green tea to treat renal stone, we examined the effect of green tea on the formation and the excretion of experimentally induced calcium oxalate (CaOx) stones in rat kidneys. MATERIALS AND METHODS: CaOx nephrolithiasis was induced by administering 1% ethylene glycol (EG) for 4 weeks. To investigate the effect of tea on the formation CaOx stones, the rats were simultaneously administered either 0.2% green tea or 0.5% rooibos tea. To verify the action of green tea on the excretion of CaOx stones, the rats were divided into four groups after the administration of 1% EG water for 4 weeks and then fed with either 0.2% green tea, 0.5% rooibos tea or 80mg/l furosemide-containing 1% EG water for 4 weeks. The right kidney was frozen for mRNA measurements, with the left fixed for counting crystal deposits. Twenty-four hour urine volume and urinary excretions of oxalate, uric acid, calcium and magnesium were measured. RESULTS: Urinary biochemistry and 24 hour urine output were apparently unchanged by taking either the green tea or rooibos tea. The increases of CaOx crystal deposits and osteopontin mRNA expressions in the kidneys by the administration of 1% EG water were markedly decreased by both tea intakes, while there were no significant differences in the mRNA levels of CuZn- and Mn-superoxide dismutases between the groups. CONCLUSIONS: Green and rooibos teas significantly attenuated the calcium crystal depositions in the kidneys. Down-regulations of the osteopontin mRNA levels may be involved in the inhibitory effects of the teas on the renal CaOx stone formation.
Animals ; Aspalathus ; Biochemistry ; Calcium Oxalate* ; Calcium* ; Ethylene Glycol ; Kidney ; Kidney Calculi ; Magnesium ; Nephrolithiasis ; Osteopontin ; Rats* ; RNA, Messenger ; Tea* ; Uric Acid ; Water

BACKGROUND: Immunohistochemical staining of skin for panaxonal marker, protein gene product 9.5 (PGP 9.5), has recently emerged as a useful diagnostic procedure for the assessment of small nerve fibers. The first aim of our study is to quantify the intraepidermal nerve fibers (IEDNF) and to investigate the histological changes of IEDNF in diabetic patents. And the second is to evaluate whether IEDNF has the correlation with the clinical, electrophysiological and biological parameters. METHODS: Skin biopsy specimens were obtained at the distal leg and proximal thigh from twelve type 2 diabetes mellitus patients having the symptoms of peripheral polyneuropathy. Fixed sections were stained using PGP 9.5, and the number of intraepidermal nerve fibers at each site was compared with those of fifteen normal healthy volunteers. We analyzed the correlation of IEDNF density with the clinical history, neuropathic symptom score (NSS), severity of diabetes (serum glucose, HbA1C), and nerve conduction studies. RESULTS: The number of IEDNF in diabetic patients was 3.0 per millimeter at the distal leg and 12.7 per millimeter at the proximal thigh compared with 15.3 per millimeter and 23.3 per millimeter in normal control respectively. Some specimen showed a morphological change of axons such as axonal swelling. The IEDNF density was not correlated with the duration of disease, biological or electrophysiological parameters but correlated with the clinical neuropathic symptom score. CONCLUSIONS: The IEDNF density is reduced in all the diabetic patients, more severely at the distal extremity. We suggest skin biopsy can provide the quantification as well as the objective evidence for the small fiber involvement of diabetic neuropathy.
Axons ; Biopsy* ; Diabetes Mellitus, Type 2 ; Diabetic Neuropathies* ; Erythromelalgia ; Extremities ; Glucose ; Healthy Volunteers ; Humans ; Leg ; Nerve Fibers* ; Neural Conduction ; Polyneuropathies ; Skin* ; Thigh

Past reports show that chronic abuse has resulted in serious sequalae of myeloneuropathy, optic neuritis and cerebral impairments. A 45-year-old male dentist developed an ascending paresthesia, severe gait ataxia, sensory loss for tactile sensation, proprioception and vibration of the four extremities. Laboratory findings revealed low serum vitamin B12 levels and high levels of homocystein. Our case adds support to previous reports that nitrous oxide could cause neurologic complications by interfering with the utilization of vitamin B12.
Dentists ; Extremities ; Gait Ataxia ; Humans ; Male ; Middle Aged ; Nitrous Oxide* ; Optic Neuritis ; Paresthesia ; Proprioception ; Sensation ; Vibration ; Vitamin B 12

PURPOSE: To evaluate whether factors related to lipid and glucose metabolism have a potential role in the progression of prostate cancer, we measured the mRNA levels of the peroxisome proliferator-activated receptor (PPAR), fatty acid elongase (ELOVL), and two glycolytic enzymes in prostate cancer (CaP) tissues. MATERIALS AND METHODS: Prostate tissues, obtained from radical prostatectomy (n=10) and transurethral resection of prostate (n=18), were quickly frozen in liquid nitrogen for RNA measurements. Transcript signals of PPAR alpha, PPAR gamma, ELOVL2, ELOVL5, phosphoglycerate kinase 1 (PgK1) and phosphoglycerate mutase 2 (PgM2) were measured using a reverse-transcription polymerase chain reaction. RESULTS: The transcript signals of PPAR alpha and PPAR gamma were down-regulated in CaP tissues. In addition, the mRNA level of PgM2 in CaP tissues was lower than that in benign prostatic hyperplasia (BPH) tissues. However, the messages for ELOVL2, ELOVL5, and PgK1 were not significantly changed. CONCLUSIONS: These results suggest that lowering of the PPARalpha, PPARgamma and PgM2 messages may be involved in aberrant and uncontrolled prostate cell growth and differentiation.
Down-Regulation* ; Glucose ; Metabolism ; Nitrogen ; Peroxisome Proliferator-Activated Receptors ; Peroxisomes* ; Phosphoglycerate Kinase ; Phosphoglycerate Mutase* ; Polymerase Chain Reaction ; PPAR alpha ; PPAR gamma* ; Prostate* ; Prostatectomy ; Prostatic Hyperplasia ; Prostatic Neoplasms* ; RNA ; RNA, Messenger ; Transurethral Resection of Prostate

STUDY DESIGN: Genetic screening of the estrogen receptor 2 (ESR2) genes in patients with ossification of the posterior longitudinal ligament (OPLL). OBJECTIVE: We studied the relationships between ESR2 gene polymorphisms and OPLL to understand the pathophysiology of OPLL. SUMMARY OF LITERATURE REVIEW: The OPLL has a strong genetic component. Several familial surveys and human leukocyte antigen (HLA) haplotype studies reveal that genetic background is an important component in the occurrence of OPLL and a large number of gene analysis studies were utilized to clarify the susceptible gene for OPLL, including COL11A2, BMP-2, TNF-alpha, NPPS, leptin receptor, transforming growth factor (TGF)-beta, Retinoic X receptor, ER, IL-1, PTH, and VDR have been performed. MATERIALS AND METHOD: Genomic deoxyribonucleic acid (DNA) samples obtained from 164 patients (93 men and 71 women) with OPLL and 219 control subjects, without the disease (105 men and 114 women) were amplified by polymerase chain reaction, and polymorphism genotypes were determined by the restriction endonuclease digestion. The distribution of genotypes was compared between the patients with the disease and the control subjects. RESULTS: The polymorphism of ESR2 [rs1256049, exon6, Val328Val, p=0.018, odd ratio (OR)=2.41, 95 confidence interval (CI)=1.15-5.02 in the recessive model] only showed statistically significant association between the control and the OPLL groups. The rest SNPs of ESR2 did not show any significant differences between the control and the OPLL groups. CONCLUSIONS: Estrogen receptor 2 (ESR2) gene polymorphisms (rs 1256049) was associated with OPLL. In future studies, we will perform target SNP chip between OPLL and candidate gene.
Digestion ; DNA ; DNA Restriction Enzymes ; Estrogen Receptor beta ; Estrogens ; Genetic Testing ; Genotype ; Haplotypes ; Humans ; Interleukin-1 ; Leukocytes ; Longitudinal Ligaments ; Male ; Ossification of Posterior Longitudinal Ligament ; Polymerase Chain Reaction ; Polymorphism, Single Nucleotide ; Receptors, Leptin ; Spine ; Succinimides ; Transforming Growth Factors ; Tumor Necrosis Factor-alpha

BACKGROUND: Risk for injuries resulting from overinflated or underinflated endotracheal tube cuff warrants adequate cuff inflation technique. Thus, this study was designed to measure the actual intracuff pressures obtained by new estimation techniques. METHODS: 95 adult surgical patients requiring tracheal intubation were randomized to two groups with respect to the endotracheal tube model: Portex(R) (n = 55) and Euromedical(R) (n = 40). After induction of anesthesia, the cuff was inflated using new estimation techniques with two different syringes: PR10 or PR20 (passive release technique using a 10-ml or 20-ml syringe, respectively). Subsequently, an aneroid manometer was used to measure the actual intracuff pressures. These inflation techniques were repeated two times. A direct cuff measurement range of 25 to 40 cmH2O was used as a reference for optimal intracuff pressure. Size 7.0 mm internal diameter (ID) tubes were used for women and size 7.5 mm ID for men. RESULTS: 88 eligible patients were studied: Portex group (n = 50) and Euromedical group (n = 38). With respect to the rate of optimal cuff inflation, PR10 was significantly higher than PR20 in both groups (56% vs. 10% in Portex group; 63.2% vs. 0% in Euromedical group, respectively) (P < 0.05). CONCLUSIONS: When direct intracuff measurement is not available, a new method, named "passive release technique" using a 10-ml syringe, is a useful alternative cuff inflation method.
Adult ; Anesthesia ; Female ; Humans ; Inflation, Economic ; Intubation ; Intubation, Intratracheal ; Male ; Statistics as Topic ; Syringes