Modified Tuoli Xiaodusan-assisted Treatment of Hp-positive Peptic Ulcer / 中国实验方剂学杂志

ObjectiveTo observe the clinical efficacy of modified Tuoli Xiaodusan （TLXDS） in adjuvant treatment of Helicobacter pylori （Hp）-positive peptic ulcer （PU） with cold-heat complex syndrome and explore its regulating effect on invasive/protective factors. MethodA total of 136 patients were randomly assigned into the control group （68 cases， including 4 cases missing， 3 cases eliminated， and 61 cases completed） and the TLXDS group （68 cases， including 4 cases missing， 1 case eliminated， and 63 cases completed）. Both groups adopted the quadruple therapy of acid suppression and Hp eradication. The patients in the control group received Weinai'an capsules orally at 4 capsules/time and 3 times/day， and those in the TLXDS group took modified TLXDS orally at 1 dose/day. The administration of both groups lasted for 8 consecutive weeks and the follow-up lasted for 12 months. Electronic gastroscopy was carried out before and after treatment for evaluating the healing of ulcer， the score of mucosal morphology， and the maturity of regenerated mucosa. The Hp infection and the score of cold-heat complex syndrome were evaluated before and after treatment. The serum levels of gastrin （GAS）， prostaglandin E2 （PGE2）， pepsinogen （PG）-Ⅰ， PG-Ⅱ， epidermal growth factor （EGF）， and trefoil factor 2 （TFF-2） were determined before and after therapy. The recurrence of Hp and PU was recorded， and the drug safety was evaluated. ResultAfter treatment， the mucosal morphology score and the traditional Chinese medicine （TCM） syndrome score in the TLXDS group were lower than those in the control group （P<0.01）. The levels of GAS， PG-Ⅰ， and PG-Ⅱ in the TLXDS group were lower than those in the control group （P<0.01）， whereas those of PGE2， EGF， and TFF-2 showed an opposite trend （P<0.01）. After treatment， the Hp eradication rate in the TLXDS group was 95.24% （60/63）， higher than that （83.61%， 51/61） in the control group （χ2=4.467， P<0.05）. The total effective rate of TCM syndromes in the TLXDS group was 98.41% （62/63）， higher than that （81.97%， 50/61） in the control group （χ2=9.589， P<0.01）. The total effective rate of the TLXDS group under gastroscopy was 98.41% （62/63）， higher than that （86.89%， 53/61） in the control group （χ2=4.525， P<0.05）. The excellent and good rate of regenerated mucosal maturity in the TLXDS group was 92.06% （58/63）， also higher than that （73.77%， 45/61） in the control group （χ2=7.372， P<0.01）. After 12 months of follow-up， the TLXDS group had lower PU recurrence rate ［19.05% （12/63） vs 37.70% （23/61）， χ2=5.325， P<0.05］ and lower Hp recurrence rate ［15.00% （9/60） vs 33.33% （17/51）， χ2=5.165， P<0.05） than the control group. No adverse reactions related to TLXDS were detected. ConclusionModified TLXDS-assisted quadruple therapy demonstrates significant short-term clinical efficacy and high Hp eradication rate for Hp-positive PU （cold-heat complex syndrome） patients. Moreover， it can adjust the levels of invasive/protective factors to facilitate ulcer healing and reduce the recurrence rates of Hp and PU in a long term， with good clinical safety.

Deep learning applied to two-dimensional color Doppler flow imaging ultrasound images significantly improves diagnostic performance in the classification of breast masses: a multicenter study / 中华医学杂志(英文版)

BACKGROUND@#The current deep learning diagnosis of breast masses is mainly reflected by the diagnosis of benign and malignant lesions. In China, breast masses are divided into four categories according to the treatment method: inflammatory masses, adenosis, benign tumors, and malignant tumors. These categorizations are important for guiding clinical treatment. In this study, we aimed to develop a convolutional neural network (CNN) for classification of these four breast mass types using ultrasound (US) images.@*METHODS@#Taking breast biopsy or pathological examinations as the reference standard, CNNs were used to establish models for the four-way classification of 3623 breast cancer patients from 13 centers. The patients were randomly divided into training and test groups (n = 1810 vs. n = 1813). Separate models were created for two-dimensional (2D) images only, 2D and color Doppler flow imaging (2D-CDFI), and 2D-CDFI and pulsed wave Doppler (2D-CDFI-PW) images. The performance of these three models was compared using sensitivity, specificity, area under receiver operating characteristic curve (AUC), positive (PPV) and negative predictive values (NPV), positive (LR+) and negative likelihood ratios (LR-), and the performance of the 2D model was further compared between masses of different sizes with above statistical indicators, between images from different hospitals with AUC, and with the performance of 37 radiologists.@*RESULTS@#The accuracies of the 2D, 2D-CDFI, and 2D-CDFI-PW models on the test set were 87.9%, 89.2%, and 88.7%, respectively. The AUCs for classification of benign tumors, malignant tumors, inflammatory masses, and adenosis were 0.90, 0.91, 0.90, and 0.89, respectively (95% confidence intervals [CIs], 0.87-0.91, 0.89-0.92, 0.87-0.91, and 0.86-0.90). The 2D-CDFI model showed better accuracy (89.2%) on the test set than the 2D (87.9%) and 2D-CDFI-PW (88.7%) models. The 2D model showed accuracy of 81.7% on breast masses ≤1 cm and 82.3% on breast masses >1 cm; there was a significant difference between the two groups (P < 0.001). The accuracy of the CNN classifications for the test set (89.2%) was significantly higher than that of all the radiologists (30%).@*CONCLUSIONS@#The CNN may have high accuracy for classification of US images of breast masses and perform significantly better than human radiologists.@*TRIAL REGISTRATION@#Chictr.org, ChiCTR1900021375; http://www.chictr.org.cn/showproj.aspx?proj=33139.

Functional organization of cytoplasmic portals controlling access to the cystic fibrosis transmembrane conductance regulator (CFTR) chloride channel pore.

The cystic fibrosis transmembrane conductance regulator (CFTR) is a Cl- channel that apparently has evolved from an ancestral active transporter. Key to the CFTR's switch from pump to channel function may have been the appearance of one or more "lateral portals." Such portals connect the cytoplasm to the transmembrane channel pore, allowing a continuous pathway for the electrodiffusional movement of Cl- ions. However, these portals remain the least well-characterized part of the Cl- transport pathway; even the number of functional portals is uncertain, and if multiple portals do exist, their relative functional contributions are unknown. Here, we used patch-clamp recording to identify the contributions of positively charged amino acid side chains located in CFTR's cytoplasmic transmembrane extensions to portal function. Mutagenesis-mediated neutralization of several charged side chains reduced single-channel Cl- conductance. However, these same mutations differentially affected channel blockade by cytoplasmic suramin and Pt(NO2)42- anions. We considered and tested several models by which the contribution of these positively charged side chains to one or more independent or non-independent portals to the pore could affect Cl- conductance and interactions with blockers. Overall, our results suggest the existence of a single portal that is lined by several positively charged side chains that interact electrostatically with both Cl- and blocking anions. We further propose that mutations at other sites indirectly alter the function of this single portal. Comparison of our functional results with recent structural information on CFTR completes our picture of the overall molecular architecture of the Cl- permeation pathway.

Identification and bioinformatic function analysis of differentially expressed miRNAs in the exosomes secreted from BEP2D cells irradiated by γ-rays / 中华放射医学与防护杂志

Objective@# To identify the differentially expressed miRNAs in the exosomes secreted from γ-ray irradiated cells and provide new clues in disclosing the mechanisms of radiation-induced bystander effects.@*Methods@#The human bronchial epithelial cells (BEP2D) were irradiated with 60Co γ-rays, and the exosomes were collected by ultracentrifugation from the culture medium of 2 Gy-irradiated cells and non-irradiated control. The exosomes were identified by an electron microscopy. The miRNA microarray technique was used to analyze the miRNA expression profiles in the exosomes. qRT-PCR was used to verify the miRNAs expression. The functional pathways of miRNAs targeting genes were predicted by informatic analysis using the databases of TargetScan, miRanda, GO and KEGG.@*Results@#Sixteen miRNA with significantly increased expression (P<0.05) were identified in the exosomes of BEP2D cells at 4 h post-2 Gy irradiation as compared with the non-irradiated control cells, among which miR-100-5p, miR-1246, miR-29b-3p, and miR-7-5p were further confirmed to be unregulated by qRT-PCR assay (P<0.05). Meanwhile, the expression changes of above-mentioned four miRNAs were also investigated in the irradiated cells. The data indicated the expression was significantly increased at 2 h post-2 Gy irradiation for the miR-100-5p, miR-1246, miR-29b-3p in addition to miR-7-5p. However, all these four miRNAs were downregulated in the cells at 4 h post-irradiation and then gradually recovered. Bioinformatics analysis showed that the targeted genes of these differentially expressed miRNAs might participate in the biological processes and signal pathways of cell adhesion, mTOR signal pathway, chromatin modification, HR and NHEJ pathways of DNA repair and so on.@*Conclusions@# Radiation-inducible miRNAs have been identified in the exosomes from the irradiated BEP2D cells. The target genes of these miRNAs play roles in a series of important biological processes and functional pathways, which provides new clues in elucidating the mechanisms of radiation-induced bystander effects.

Study on Separation and Purification of Total Flavonoids from Polygonum hydropiper Linn. by Macroporous Resin / 中国中医药信息杂志

Objective To study the separation and purification of total flavonoids from Polygonum hydropiper Linn. by macroporous adsorption resin; To investigate the effects of various factors in the process of static and dynamic adsorption on effects of adsorption and desorption to determine the optimum capability. Methods The total flavonoids from Polygonum hydropiper Linn. was extracted; the contents of flavonoids of ethyl acetate part (FEA) and flavonoids of n-butanol part (FNB) were detected; the adsorption experiment of FEA and FNB was conducted by macroporous adsorption resin. D101, AB-8, DM130, and XDA-8 macroporous resin were used to optimize the separation and purification process. Results The XDA-8 macroporous resin was selected to enrich total flavonoids from Polygonum hydropiper Linn. The optimum process was: adjusting the pH value of FEA flavonoids to 6, the concentration of sample was 750 g/mL, eluting with 75% ethanol to elute 5 bed volumes at a flow rate of 1 bed per hour; adjusting the pH value of FNB flavonoids to 6, the concentration of sample was 1 mg/mL, eluting with 60% ethanol to elute 5 bed volumes at a flow rate of 1 bed per hour. Conclusion XDA-8 macroporous adsorption resin is helpful to separate and purify the total flavonoids of Polygonum hydropiper Linn.by the best process.

Identification and analysis of exosomal miRNAs in human umbilical vein endothelial cells irradiated by 60 Co γ-rays / 军事医学

Objective To study the changes in miRNAs expression in the exosomes of human umbilical vein endothelial cells(HUVECs) after 60 Co γ-rays expose using microRNA(miRNA) chips and bioinformatics techniques so as to provide new clues to the mechanism of radiation-induced vascular tissue injury and its bystander effects.Methods HUVECs exosomes were collected in the control and 4 Gy irradiated cells by ultra-high-speed centrifugation,and further confirmed using transmission electron microscopy (TEM) and Western blotting of exosomes biomarkers.miRNA microarray was used to analyze miRNA expression profiles of exosomes and cells.Also,real-time quantitative PCR(qRT-PCR) was used to verify differentially expressed miRNAs,and the miRDB and TargetScan were performed to predict the target genes of the differentially expressed miRNAs.Bioinformatics analysis was performed using DAVID,KEGG and other online tools.Results Compared with the control exosomes from non-irradiated HUVECs,miRNA microarray analysis revealed that 5 up-regulated,and 13 down-regulated miRNAs were identified in the exosomes from HUVECs at 0.5 h after 4 Gy-irradiation,and 16 up-regulated and 5 down-regulated miRNAs at 2 h after 4 Gy-irradiation.Moreover,38 and 85 miRNAs were differentially expressed respectively in the HUVECs at 0.5 h and 2 h after radiation.The difference was statistically significant(P<0.01).The results of bioinformatics showed that these miRNAs might exert the radiation-induced bystander effect (RIBE) by regulating MAPK signal pathways,RAS and PI3K-Akt signal pathways.Conclusion The ionizing radiation injury significantly alters the components and expression levels of exosomal miRNAs,which play important roles in regulating the signal pathways in response to radiation.

Effects of calcium elevation on intracellular oxidative stress in human lens epithelial cells SRA01/04 / 眼科新进展

Objective To investigate the influence of calcium elevation on oxida tive stress in human lens epithelial cells (HLEC) SRA01/04.Method The cells (2 x 103 cells/well) which in the period of logarithmic phase were seeded into 96-well plates with three replicates for the two groups;and in the experimental group,SRA01/04 cells were exposed to a CaCI2 concentration gradient (3 mmol · L-1,5 mmol · L-1,7 mmol · L-1,9 mmol · L-1,11 mmol · L-1,13 mmol · L-1,15 mmol · L-1,17 mmol · L-1,19 mmol · L-1) for 0 h,12 h,24 h,36 h;while the cells in the control group were cultured in complete 1640 medium.Cell counting kit-8 (CCK-8) assay was used to measure cell viability.The levels of intracellular superoxide dismutase (SOD),glutathione (GSH) content and oxidized glutathione (GSSG) / total glutathione (T-GSH) were determined by using the microplate-reader method with the commercial total/oxidized glutathione and sod quantification kit.Results At first,the survival rate of SRA01/04 cells treated with 3 mmol · L-1,5 mmol · L-1,7 mmol· L-1 CaCL2 for 24 h showed a significant decrease with the increase of CaCl2 concentration by CCK-8 assays,but gradually increased when the concentration increased to 9 mmol · L-1,and the difference approached statistical significance (P ＜ 0.05).Meanwhile,there was significant difference in the viability of the control group (0.592 + 0.055) and cells exposed to 15 mmol · L-1 CaCI2 (0.293 + 0.02) (t =7.811,P ＜0.05).Cell treatment with 15 mmol· L-1 CaC12 for 24 h was the most appropriate condition for HLEC apoptosis,followed by the appearance of nuclear fragmentation and dissolution,enhanced intracellular SOD viability (t =-6.417,P ＜ 0.05),decreased T-GSH content (t =13.816,P ＜ 0.05),and increased ratio of GSSG/T-GSH (t =-4.396,P ＜ 0.05) when compared with the control group,and the differences were statistically significant.Conclusion Intracellular calcium elevation can inhibit the cell viability and increase the levels of SOD and GSSG in HLEC to aggravate the intracellular oxidative damage.

Effects of calcium elevation on intracellular oxidative stress in human lens epithelial cells SRA01/04 / 眼科新进展

Objective To investigate the influence of calcium elevation on oxida tive stress in human lens epithelial cells (HLEC) SRA01/04.Method The cells (2 x 103 cells/well) which in the period of logarithmic phase were seeded into 96-well plates with three replicates for the two groups;and in the experimental group,SRA01/04 cells were exposed to a CaCI2 concentration gradient (3 mmol · L-1,5 mmol · L-1,7 mmol · L-1,9 mmol · L-1,11 mmol · L-1,13 mmol · L-1,15 mmol · L-1,17 mmol · L-1,19 mmol · L-1) for 0 h,12 h,24 h,36 h;while the cells in the control group were cultured in complete 1640 medium.Cell counting kit-8 (CCK-8) assay was used to measure cell viability.The levels of intracellular superoxide dismutase (SOD),glutathione (GSH) content and oxidized glutathione (GSSG) / total glutathione (T-GSH) were determined by using the microplate-reader method with the commercial total/oxidized glutathione and sod quantification kit.Results At first,the survival rate of SRA01/04 cells treated with 3 mmol · L-1,5 mmol · L-1,7 mmol· L-1 CaCL2 for 24 h showed a significant decrease with the increase of CaCl2 concentration by CCK-8 assays,but gradually increased when the concentration increased to 9 mmol · L-1,and the difference approached statistical significance (P ＜ 0.05).Meanwhile,there was significant difference in the viability of the control group (0.592 + 0.055) and cells exposed to 15 mmol · L-1 CaCI2 (0.293 + 0.02) (t =7.811,P ＜0.05).Cell treatment with 15 mmol· L-1 CaC12 for 24 h was the most appropriate condition for HLEC apoptosis,followed by the appearance of nuclear fragmentation and dissolution,enhanced intracellular SOD viability (t =-6.417,P ＜ 0.05),decreased T-GSH content (t =13.816,P ＜ 0.05),and increased ratio of GSSG/T-GSH (t =-4.396,P ＜ 0.05) when compared with the control group,and the differences were statistically significant.Conclusion Intracellular calcium elevation can inhibit the cell viability and increase the levels of SOD and GSSG in HLEC to aggravate the intracellular oxidative damage.

A New Method for Discriminating between Bronchial and Pulmonary Arterial Phases using Contrast-Enhanced Ultrasound.

This study aimed to explore the value of a real-time comparative observation method using contrast-enhanced ultrasound (CEUS) for discriminating between bronchial and pulmonary arterial phases in diagnosing lung diseases. Forty-nine patients with 50 pulmonary lesions (45 peripheral lesions and five central lesions with obstructive atelectasis, including 36 malignant tumors, five tuberculomas, four inflammatory pseudotumors and five pneumonia lesions) detected via computed tomography and visible on ultrasonography were enrolled in this study. The arterial phases were determined by comparing contrast agent arrival time (AT) in the peripheral lung lesion with that in adjacent lung tissue, referred to as a real-time comparative observation method. Detection rates of this observation method were 100% (50/50) for pulmonary arterial phase and 88% (44/50) for bronchial arterial phase. Using the instrument's built-in graphing and analysis software, a time-intensity curve was constructed based on a chosen region of interest within the lesion where enhancement was the most obvious. Commonly used perfusion indicators in CEUS, such as AT, time-to-peak and peak intensity, were obtained from the time-intensity curve. Percutaneous puncture biopsies were performed under ultrasound guidance, and specimens of all 50 lesions were examined pathologically. AT was significantly shorter in patients with pneumonia than in those with malignant tumors or chronic inflammation (p < 0.05), whereas no difference was seen between those with malignant tumors and those with chronic inflammation. No significant differences in time-to-peak or peak intensity were seen among those with various lung diseases (p > 0.05). This is the first description of a real-time comparative observation method using CEUS for determining the arterial phases in the lungs. This method is accurate, simple to perform and provides a direct display. It is expected to become a practical and feasible tool for diagnosing lung diseases.

Nanostructures of Indium Gallium Nitride Crystals Grown on Carbon Nanotubes.

Nanostructure (NS) InGaN crystals were grown on carbon nanotubes (CNTs) using metalorganic chemical vapor deposition. The NS-InGaN crystals, grown on a ~5-µm-long CNT/Si template, were estimated to be ~100-270 nm in size. Transmission electron microscope examinations revealed that single-crystalline InGaN NSs were formed with different crystal facets. The observed green (~500 nm) cathodoluminescence (CL) emission was consistent with the surface image of the NS-InGaN crystallites, indicating excellent optical properties of the InGaN NSs on CNTs. Moreover, the CL spectrum of InGaN NSs showed a broad emission band from 490 to 600 nm. Based on these results, we believe that InGaN NSs grown on CNTs could aid in overcoming the green gap in LED technologies.

Pseudohalide anions reveal a novel extracellular site for potentiators to increase CFTR function.

BACKGROUND AND PURPOSE: There is great interest in the development of potentiator drugs to increase the activity of the cystic fibrosis transmembrane conductance regulator (CFTR) in cystic fibrosis. We tested the ability of several anions to potentiate CFTR activity by a novel mechanism. EXPERIMENTAL APPROACH: Patch clamp recordings were used to investigate the ability of extracellular pseudohalide anions (Co(CN)(6) (3-) , Co(NO(2) )(6) (3-) , Fe(CN)(6) (3-) , IrCl(6) (3-) , Fe(CN)(6) (4-) ) to increase the macroscopic conductance of mutant CFTR in intact cells via interactions with cytoplasmic blocking anions. Mutagenesis of CFTR was used to identify a possible molecular mechanism of action. Transepithelial short-circuit current recordings from human airway epithelial cells were used to determine effects on net anion secretion. KEY RESULTS: Extracellular pseudohalide anions were able to increase CFTR conductance in intact cells, as well as increase anion secretion in airway epithelial cells. This effect appears to reflect the interaction of these substances with a site on the extracellular face of the CFTR protein. CONCLUSIONS AND IMPLICATIONS: Our results identify pseudohalide anions as increasing CFTR function by a previously undescribed molecular mechanism that involves an interaction with an extracellular site on the CFTR protein. Future drugs could utilize this mechanism to increase CFTR activity in cystic fibrosis, possibly in conjunction with known intracellularly-active potentiators.

Mitochondria couple cellular Ca(2+) signal transduction / 生理学报

It has been shown that mitochondria not only control their own Ca(2+) concentration ([Ca(2+)]), but also exert an influence over Ca(2+) signaling of the entire cell, including the endoplasmic reticulum or the sarcoplasmic reticulum, the plasma membrane, and the nucleus. That is to say, mitochondria couple cellular metabolic state with Ca(2+) transport processes. This review focuses on the ways in which the mitochondrial Ca(2+) handling system provides integrity and modulation for the cell to cope with the complex actions throughout its life cycle, enumerates some indeterminate aspects about it, and finally, prospects directions of future research.

Functional differences in pore properties between wild-type and cysteine-less forms of the CFTR chloride channel.

Studies of the structure and function of the cystic fibrosis transmembrane conductance regulator (CFTR) Cl(-) channel have been advanced by the development of functional channel variants in which all 18 endogenous cysteine residues have been mutated ("cys-less" CFTR). However, cys-less CFTR has a slightly higher single-channel conductance than wild-type CFTR, raising questions as to the suitability of cys-less as a model of the wild-type CFTR pore. We used site-directed mutagenesis and patch-clamp recording to investigate the origin of this conductance difference and to determine the extent of functional differences between wild-type and cys-less CFTR channel permeation properties. Our results suggest that the conductance difference is the result of a single substitution, of C343: the point mutant C343S has a conductance similar to cys-less, whereas the reverse mutation, S343C in a cys-less background, restores wild-type conductance levels. Other cysteine substitutions (C128S, C225S, C376S, C866S) were without effect. Substitution of other residues for C343 suggested that conductance is dependent on amino acid side chain volume at this position. A range of other functional pore properties, including interactions with channel blockers (Au[CN] (2) (-) , 5-nitro-2-[3-phenylpropylamino]benzoic acid, suramin) and anion permeability, were not significantly different between wild-type and cys-less CFTR. Our results suggest that functional differences between these two CFTR constructs are of limited scale and scope and result from a small change in side chain volume at position 343. These results therefore support the use of cys-less as a model of the CFTR pore region.

Regulation of CFTR chloride channel macroscopic conductance by extracellular bicarbonate.

The CFTR contributes to Cl⁻ and HCO3⁻ transport across epithelial cell apical membranes. The extracellular face of CFTR is exposed to varying concentrations of Cl⁻ and HCO3⁻ in epithelial tissues, and there is evidence that CFTR is sensitive to changes in extracellular anion concentrations. Here we present functional evidence that extracellular Cl⁻ and HCO3⁻ regulate anion conduction in open CFTR channels. Using cell-attached and inside-out patch-clamp recordings from constitutively active mutant E1371Q-CFTR channels, we show that voltage-dependent inhibition of CFTR currents in intact cells is significantly stronger when the extracellular solution contains HCO3⁻ than when it contains Cl⁻. This difference appears to reflect differences in the ability of extracellular HCO3⁻ and Cl⁻ to interact with and repel intracellular blocking anions from the pore. Strong block by endogenous cytosolic anions leading to reduced CFTR channel currents in intact cells occurs at physiologically relevant HCO3⁻ concentrations and membrane potentials and can result in up to ∼50% inhibition of current amplitude. We propose that channel block by cytosolic anions is a previously unrecognized, physiologically relevant mechanism of channel regulation that confers on CFTR channels sensitivity to different anions in the extracellular fluid. We further suggest that this anion sensitivity represents a feedback mechanism by which CFTR-dependent anion secretion could be regulated by the composition of the secretions themselves. Implications for the mechanism and regulation of CFTR-dependent secretion in epithelial tissues are discussed.

Regulation of conductance by the number of fixed positive charges in the intracellular vestibule of the CFTR chloride channel pore.

Rapid chloride permeation through the cystic fibrosis transmembrane conductance regulator (CFTR) Cl(-) channel is dependent on the presence of fixed positive charges in the permeation pathway. Here, we use site-directed mutagenesis and patch clamp recording to show that the functional role played by one such positive charge (K95) in the inner vestibule of the pore can be "transplanted" to a residue in a different transmembrane (TM) region (S1141). Thus, the mutant channel K95S/S1141K showed Cl(-) conductance and open-channel blocker interactions similar to those of wild-type CFTR, thereby "rescuing" the effects of the charge-neutralizing K95S mutation. Furthermore, the function of K95C/S1141C, but not K95C or S1141C, was inhibited by the oxidizing agent copper(II)-o-phenanthroline, and this inhibition was reversed by the reducing agent dithiothreitol, suggesting disulfide bond formation between these two introduced cysteine side chains. These results suggest that the amino acid side chains of K95 (in TM1) and S1141 (in TM12) are functionally interchangeable and located closely together in the inner vestibule of the pore. This allowed us to investigate the functional effects of increasing the number of fixed positive charges in this vestibule from one (in wild type) to two (in the S1141K mutant). The S1141K mutant had similar Cl(-) conductance as wild type, but increased susceptibility to channel block by cytoplasmic anions including adenosine triphosphate, pyrophosphate, 5-nitro-2-(3-phenylpropylamino)benzoic acid, and Pt(NO(2))(4)(2-) in inside-out membrane patches. Furthermore, in cell-attached patch recordings, apparent voltage-dependent channel block by cytosolic anions was strengthened by the S1141K mutation. Thus, the Cl(-) channel function of CFTR is maximal with a single fixed positive charge in this part of the inner vestibule of the pore, and increasing the number of such charges to two causes a net decrease in overall Cl(-) transport through a combination of failure to increase Cl(-) conductance and increased susceptibility to channel block by cytosolic substances.

The Influence of Age and Prostate Volume on the Cancer Detection Rate in Korean Men with PSA Levels of 4.0 to 10.0ng/ml: a Multicenter, Retrospective Study / 대한비뇨기과학회지

PURPOSE: We retrospectively investigated how patients age and prostate volume influence on the cancer detection rate in Korean men with prostate-specific antigen (PSA) levels of 4.0 to 10.0ng/ml. MATERIALS AND METHODS: 791 Korean men who underwent transrectal ultrasound guided prostate biopsies (TRBx) at 12 medical centers were analyzed retrospectively during the previous 10 years. TRBx were performed in cases with PSA levels of 4.0 to 10.0ng/ml. The biopsy-proven cancer patient group was compared to the non-cancer patient group according to age, PSA, prostate volume and PSAD. RESULTS: Among the 791 patients who underwent TRBx, prostate cancer was detected in 123 patients (15.5%). The mean age (cancer group vs non-cancer group=69.1 vs 63.8 year-old), prostate volume (38.0 vs 42.5ml, respectively) and PSAD (0.21 vs 0.18ng/ml/ml, respectively) were found in statistically significant between the two groups. The cancer detection rate (20.1%) in the small prostate (less than 40ml) was significantly higher than that (10.3%) of the large prostate. The cancer detection rate was significantly increased with age: from 14.4% for the 50 to 59 year-old patients to 31.6% for the 80 or more year-old patients. CONCLUSIONS: The cancer detection rate in Korean men with a gray zone PSA level is lower than that of Caucasians. However, regarding the detection of prostate cancer in Korean men, the older age group and the patients with less than 40ml of prostate volume among the patients with gray zone PSA levels are considered as the important factors to decide whether biopsy of prostate is needed.

A Multicenter Study of the Detection Rate for Prostate Cancer according to the Serum Prostate-Specific Antigen Level in Korean Men / 대한비뇨기과학회지

PURPOSE: The incidence of prostate cancer is increasing in Korea, but compared with western counties, the incidence is relatively low. The detection rate of prostate cancer, according to the serum prostate-specific antigen (PSA) level, is reportedly different in Korean men, but this remains to be confirmed. We retrospectively reviewed the data of prostate biopsies, and evaluated the detection rate of prostate cancer from biopsies, according to the serum PSA level in Korean men. MATERIALS AND METHODS: We retrospectively reviewed the results of 2,422 Korean men who had undergone prostate biopsies at 12 medical centers. Prostate biopsies were performed in cases of high PSA levels, greater than 4ng/ml, or abnormal findings on digital rectal examination. RESULTS: Of the 2,422 men, 39.7% had a positive biopsy. With PSA levels between 4 and 10ng/ml, the detection rate of prostate cancer was 15.9%. This rate was similar to that of the Japanese (15.8%), but quite different from that of American men (25%). With PSA levels above 10ng/ml, 59.5% of men had a positive biopsy. For PSA levels > or= 4ng/ml and > or= 10ng/ml, the detection rates were 42.1 and 59.5%, respectively. CONCLUSIONS: When the serum PSA levels were divided into 4 subdivisions (4.0-10.0, 10.0-20.0 and 20.0-100.0ng/ml and more than 100.0ng/ml), the detection rates were 15.9, 34.1, 66.2 and 93.8%, respectively.

[Pregnanolone effects on the blood pressure of stress-induced hypertension in rats].

This study was conducted to determine the effect of pregnanolone (PGN) on blood pressure of a rat model of stress-induced hypertension (SIH). This model was established by applying electric shock to animal feet together with noise. PGN was administered intraperitoneally at 0.24 mg/kg.d(-1) and blood pressure, angiotensin II (Ang II) levels, and the expression of Fos-like protein immunoreactive (FLI) neurons in brain areas were determined. Rats were randomly divided into five groups: (1) control, (2) stressed for 1 h, (3) stressed for 1 h after PGN pretreatment, (4) stressed for a 2 h session, twice a day, for 15 d, and (5) stressed for a 2 h session after PGN pretreatment, twice a day, for 15 d. The results showed that increased systolic pressure of tail artery caused by a 15-d stress treatment was significantly reduced by PGN pretreatment (P<0.001). Ang II levels, measured by radioactive immunoreactivity, were significantly elevated (P<0.001) after the rats were stressed for 1 h or 15 d, the Ang II level was significantly reduced by PGN treatment in both 1 h and 15 d stress groups (P<0.05). Only a small number of FLI neurons were found in the brain areas of the control group, 15 d stress group, and 15 d stress with PGN group. In the 1 h stress group, more FLI neurons were found in the lateral habenular nucleus, the medial habenular nucleus, the paraventricular nucleus, the central nucleus of amgydaloid and the lateral hypothalamus compared with the control group. PGN pretreatment significantly prevented the increase in the number of FLI neurons. These results indicate that PGN pretreatment prevents elevation of tail artery systolic pressure in SIH rats and that this effect of PGN may be mediated through reducing Ang II level and inhibiting the activity of cardiovascular center involved in stress.

Pregnanolone effects on the blood pressure of stress-induced hypertension in rats / 生理学报

This study was conducted to determine the effect of pregnanolone (PGN) on blood pressure of a rat model of stress-induced hypertension (SIH). This model was established by applying electric shock to animal feet together with noise. PGN was administered intraperitoneally at 0.24 mg/kg.d(-1) and blood pressure, angiotensin II (Ang II) levels, and the expression of Fos-like protein immunoreactive (FLI) neurons in brain areas were determined. Rats were randomly divided into five groups: (1) control, (2) stressed for 1 h, (3) stressed for 1 h after PGN pretreatment, (4) stressed for a 2 h session, twice a day, for 15 d, and (5) stressed for a 2 h session after PGN pretreatment, twice a day, for 15 d. The results showed that increased systolic pressure of tail artery caused by a 15-d stress treatment was significantly reduced by PGN pretreatment (P<0.001). Ang II levels, measured by radioactive immunoreactivity, were significantly elevated (P<0.001) after the rats were stressed for 1 h or 15 d, the Ang II level was significantly reduced by PGN treatment in both 1 h and 15 d stress groups (P<0.05). Only a small number of FLI neurons were found in the brain areas of the control group, 15 d stress group, and 15 d stress with PGN group. In the 1 h stress group, more FLI neurons were found in the lateral habenular nucleus, the medial habenular nucleus, the paraventricular nucleus, the central nucleus of amgydaloid and the lateral hypothalamus compared with the control group. PGN pretreatment significantly prevented the increase in the number of FLI neurons. These results indicate that PGN pretreatment prevents elevation of tail artery systolic pressure in SIH rats and that this effect of PGN may be mediated through reducing Ang II level and inhibiting the activity of cardiovascular center involved in stress.