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Monovalent salts are generally believed to stabilize DNA duplex by weakening inter-strand electrostatic repulsion. Unexpectedly, our force-induced hairpin unzipping experiments and thermal melting experiments show that LiCl, NaCl, KCl, RbCl, and CsCl at concentrations beyond ~1 M destabilize DNA, RNA, and RNA-DNA duplexes. The two types of experiments yield different changes in free energy during melting, while the results that high concentration monovalent salts destabilize duplexes are common. The effects of these monovalent ions are similar but also have noticeable differences. From 1 M to 4 M, DNA duplex is destabilized by about 0.3 k B T/bp and the melting temperature decreases by about 10 o C. Our all-atom simulations reveal this effect is caused by overcharging, where excessive ion absorption inverts the effective DNA charge from negative to positive. Furthermore, our coarse-grained simulations obtain a phase diagram that indicates whether DNA overcharging occurs at a given cation valence and concentration. These findings challenge the traditional belief that DNA overcharging occurs only with multivalent ions and have significant implications for polyelectrolyte theory, DNA nanomaterials, DNA nanotechnology, and DNA biophysics., Competing Interests: Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

Background: Accurate microsatellite instability (MSI) testing is essential for identifying gastric cancer (GC) patients eligible for immunotherapy. We aimed to develop and validate a CT-based radiomics signature to predict MSI and immunotherapy outcomes in GC., Methods: This retrospective multicohort study included a total of 457 GC patients from two independent medical centers in China and The Cancer Imaging Archive (TCIA) databases. The primary cohort (n = 201, center 1, 2017-2022), was used for signature development via Least Absolute Shrinkage and Selection Operator (LASSO) and logistic regression analysis. Two independent immunotherapy cohorts, one from center 1 (n = 184, 2018-2021) and another from center 2 (n = 43, 2020-2021), were utilized to assess the signature's association with immunotherapy response and survival. Diagnostic efficiency was evaluated using the area under the receiver operating characteristic curve (AUC), and survival outcomes were analyzed via the Kaplan-Meier method. The TCIA cohort (n = 29) was included to evaluate the immune infiltration landscape of the radiomics signature subgroups using both CT images and mRNA sequencing data., Results: Nine radiomics features were identified for signature development, exhibiting excellent discriminative performance in both the training (AUC: 0.851, 95%CI: 0.782, 0.919) and validation cohorts (AUC: 0.816, 95%CI: 0.706, 0.926). The radscore, calculated using the signature, demonstrated strong predictive abilities for objective response in immunotherapy cohorts (AUC: 0.734, 95%CI: 0.662, 0.806; AUC: 0.724, 95%CI: 0.572, 0.877). Additionally, the radscore showed a significant association with PFS and OS, with GC patients with a low radscore experiencing a significant survival benefit from immunotherapy. Immune infiltration analysis revealed significantly higher levels of CD8 + T cells, activated CD4 + B cells, and TNFRSF18 expression in the low radscore group, while the high radscore group exhibited higher levels of T cells regulatory and HHLA2 expression., Conclusion: This study developed a robust radiomics signature with the potential to serve as a non-invasive biomarker for GC's MSI status and immunotherapy response, demonstrating notable links to post-immunotherapy PFS and OS. Additionally, distinct immune profiles were observed between low and high radscore groups, highlighting their potential clinical implications., (© 2024. The Author(s).)

We propose a scheme to achieve nonreciprocal magnon blockade via the Barnett effect in a magnon-based hybrid system. Due to the rotating yttrium iron garnet (YIG) sphere, the Barnett shift induced by the Barnett effect can be tuned from positive to negative via controlling magnetic field direction, leading to nonreciprocity. We show that a nonreciprocal unconventional magnon blockade (UMB) can emerge only from one magnetic field direction but not from the other side. Particularly, by further tuning system parameters, we simultaneously observe a nonreciprocal conventional magnon blockade (CMB) and a nonreciprocal UMB. This result achieves a switch between efficiency (UMB) and purity (CMB) of a single-magnon blockade. Interestingly, stronger UMB can be reached under stronger qubit-magnon coupling, even the strong coupling regime. Moreover, the nonreciprocity of the magnon blockade is sensitive to temperature. This work opens up a way for achieving quantum nonreciprocal magnetic devices and chiral magnon communications.

The mammalian target of rapamycin (mTOR) has been proved to be an effective target for cancer therapy. Two kinds of mTOR inhibitors, the rapalogs and mTOR kinase inhibitors (TORKi), have been developed and clinically validated in several types of malignancies. Compared with rapalogs, TORKi can exert better antitumor activity by inhibiting both mTORC1 and mTORC2, but the clinical development of current TORKi candidates has been relative slow, more TORKi with novel scaffold need to be developed to expand the current pipelines. In this study, a series of 9-methyl-9H-purine and thieno[3, 2-d]pyrimidine derivatives were designed, synthesized and biological evaluation. Most of these compounds exhibited good mTOR kinase inhibitory activity and selectivity over PI3Kα. Subsequent antiproliferative assay allowed us to identify the lead compound 15i, which display nanomolar to low micromolar IC 50 s against six human cancer cell lines. 15i could induce cell cycle arrest of MCF-7, PC-3 and A549 cells at the G0/G1 phase and suppress the migration and invasion of these cancer cells by suppressing the phosphorylation of AKT and P70S6 kinase. It could also regulate autophagy-related proteins to induce autophagy. Therefore, 15i would be a starting point for the development of new TORKi as anticancer drug., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Inc. All rights reserved.)

Background: In this study, we aimed to investigate the role of miR-26a and miR-26b in the management of AF., Methods: Real-time PCR was carried out to determine plasma microRNA expression in AF patients pre- and post-radiofrequency ablation. The correlation between the expression of SELP and miR-26a/miR-26b was also studied using luciferase assays to establish a miR-26a/miR-26b/SELP signaling pathway., Results: The relative expression of SELP reached its peak in pre-ablation AF ( +) patients, while ablation treatment reduced the expression of SELP in AF ( +) patients. Similarly, AF pigs showed dysregulation of miR-26a/b and SELP, thus verifying the involvement of miR-26a/b and SELP in AF. Meanwhile, the regulatory association between SELP and miR-26a/b was also investigated, and the results showed that the presence of pre-miR-26a/b increased the levels of miR-26a/b and inhibited the mRNA/protein expression of SELP. Finally, using bioinformatic tools and luciferase assays, SELP mRNA was confirmed as the target of miR-26a/b, which affected the effect of AF ablation treatment., Conclusions: RFA helped to restore circulating levels of miR-26, which were reduced in atrial fibrillation. Meanwhile, miR-26 is a potential cause for the elevated plasma levels of pro-thrombogenic SELP in that disease., (© 2022. The Author(s).)

This study investigatedthe different effects of long-term glucocorticoid (GC) interventions on the microarchitectures of cortical and cancellous bones of the femoral head. Eighteen female skeletal mature sheep were randomly allocated into 3 groups, 6 each. Group 1 received prednisolone interventions (0.60 mg/kg/day, 5 times weekly) for 7 months. Group 2 received the same interventions as Group 1 and then further observed 3 months without interventions. Control Group was left nonintervention. After killing the animals, all femoral heads were scanned by micro-CT to determine their microstructural properties. In cancellous bone of femoral head, GC interventions led to significant decrease of bone volume fraction, trabecular thickness, trabecular separation, but increase of structure model index and bone surface density (p < 0.05). While in cortical bone, there were no differences between the Group 1 and in microstructural properties (p > 0.05) except greater trabecular thickness in the control group. In addition, three months after cessation of glucocorticoid interventions, most microstructural properties of cancellous bone were significant reversed, but not cortical thickness of femoral head. In contrast to cancellous bone, the microarchitectures of cortical bone were not changed obviously after long-term GC interventions.

This prospective study was designed to examine the combined influence of insulin resistance (IR) and inflammatory biomarker levels on type 2 diabetes mellitus (T2DM) among 1,903 Inner Mongolians. During follow-up, 205 (10.77%) participants developed T2DM, and the incidence of T2DM was higher among subjects with IR, elevated C-reactive protein (CRP), elevated sICAM-1, elevated sE-selectin, or the coexistences of IR with elevated CRP, elevated sICAM-1, elevated sE-selectin, and elevated angiotensin II (all P < 0.05) compared with patients without IR or any elevated biomarkers. In multivariate analysis, the odd ratios [OR, (95% confidence intervals)] for these conditions were 1.944 (1.405-2.691), 2.003 (1.449-2.767), 1.706 (1.232-2.362), 1.560 (1.123-2.165), 2.708 (1.809-4.054), 1.885 (1.155-3.078), 2.101 (1.340-3.295), and 2.260 (1.426-3.582), respectively. Our findings demonstrated that IR and elevated inflammatory biomarkers were associated with T2DM, and that the coexistence of IR and elevated inflammatory biomarkers increased the risk of T2DM., (Copyright © 2018 The Editorial Board of Biomedical and Environmental Sciences. Published by China CDC. All rights reserved.)

The expansion of built-up area will cause stress effect on the regional natural ecological security pattern during urbanization process. Taking rapid expanding regions of four inland and coastal cities as study areas, including Tongzhou in Beijing, Zhengding in Hebei, Tanggu in Tianjin and Xiamen in Fujian, we constructed regional landscape stress indexes according to the principle of landscape ecology and comparatively analyzed the landscape pattern characteristics of rapid expanding regions and the differences of stress effect of artificial landscapes on four natural landscapes ecological security pattern in the process of rapid urbanization. Results showed that landscape erosion indexes of Tongzhou, Zhengding, Tanggu and Xiamen in 2015 were 1.039, 0.996, 1.239 and 0.945, respectively, which indicated that the natural landscapes were eroded significantly. Natural landscape types of those four regions presented different threatened levels. Among all natural landscape types, unused land and waters were worst threatened in Tongzhou, Zhengding and Tanggu, while in Xiamen cultivated land and waters showed the highest threat levels. The waters threat indexes of those four areas were all more than 0.743. Landscape isolation indexes of waters and unused land of the inland cities were greater than those of coastal cities, which meant water distribution of inland cities in the space was less gathered than that of coastal cities. Besides, compared with the other natural landscape, unused land and waters suffered the largest stress from artificial landscapes.

Objective: No previous studies have evaluated the association between dyslipidemia, alcohol drinking, and diabetes in an Inner Mongolian population. We aimed to evaluate the co-effects of drinking and dyslipidemia on diabetes incidence in this population., Methods: The present study was based on 1880 participants from a population-based prospective cohort study among Inner Mongolians living in China. Participants were classified into four subgroups according to their drinking status and dyslipidemia. Multivariate logistic regression analysis and receiver operating characteristic (ROC) curves were used to evaluate the association between alcohol drinking, dyslipidemia, and diabetes., Results: During the follow-up period, 203 participants were found to have developed diabetes. The multivariable-adjusted odds ratios (95% confidence interval) for the incidence of non-dyslipidemia/drinkers, dyslipidemia/non-drinkers, and dyslipidemia/drinkers in diabetic patients were 1.40 (0.82-2.37), 1.73 (1.17-2.55), and 2.31 (1.38-3.87), respectively, when compared with non-dyslipidemia/non-drinkers. The area under the ROC curve for a model containing dyslipidemia and drinking status along with conventional factors (AUC=0.746) was significantly (P=0.003) larger than the one containing only conventional factors (AUC=0.711)., Conclusion: The present study showed that dyslipidemia was an independent risk factor for diabetes, and that drinkers with dyslipidemia had the highest risk of diabetes in the Mongolian population. These findings suggest that dyslipidemia and drinking status may be valuable in predicting diabetes incidence., (Copyright © 2016 The Editorial Board of Biomedical and Environmental Sciences. Published by China CDC. All rights reserved.)

We aimed to investigate the cumulative effect of overweight and resting heart rate on prediabetes/diabetes incidence in an 10-year follow-up study in Inner Mongolians. Among 1729 participants who were free from prediabetes and diabetes at baseline, 503 and 155 subjects developed prediabetes and diabetes, respectively. We categorized the participants into 4 subgroups according to overweight and resting heart rate status. The multivariate-adjusted OR (95% CI) in normal weight with heart rate ≥80 bpm, overweight with heart rate <80 bpm, and overweight with heart rate ≥80 bpm were 1.24 (0.95-1.61), 1.83 (1.29-2.61), 2.20 (1.41-3.45) for prediabetes and 1.52 (0.97-2.40), 3.64 (2.21-6.01), 4.61 (2.47-8.61) for diabetes, respectively, compared with normal weight with heart rate <80 bpm. The area under ROC curve (AUC) for the prediction of diabetes incidence for a model containing overweight and resting heart rate, along with conventional factors (AUC = 0.751), was significantly (P = 0.003) larger than the one containing only conventional factors (AUC = 0.707). Our study indicated that overweight was an independent risk factor of prediabetes and diabetes, and overweight with faster resting heart rate might further increase the risk of prediabetes and diabetes.

Stabilization of heavy metals in situ was investigated. Two combined amendments (LS, limestone+sepiolite; HZ, hydroxyhistidine+zeolite) were applied at ratios of 0.2%, 0.4%, and 0.8% (w/w) to paddy soil with multi-metal (Pb, Cd, Cu, and Zn) contamination. The effects of these two combined amendments on heavy metal bioavailability in soil, and on uptake and accumulation of heavy metals in rice plants were investigated. Application of LS and HZ significantly increased soil pH values and cation exchange capacity contents, and resulted in a reduction in exchangeable fraction of metals and in extract metal concentrations of amended soils through toxicity characteristic leaching procedure (TCLP). LS and HZ obviously inhibited uptake and accumulation of Pb, Cd, Cu, and Zn in rice plants. Compared with the control soil, concentrations of Pb, Cd, Cu, and Zn in brown rice were decreased by 10.6-31.8%, 16.7-25.5%, 11.5-22.1%, and 11.7-16.3%, respectively, as a result of 0.2% to 0.8% addition of LS, and decreased by 5.1-40.8%, 16.7-20.0%, 8.1-16.2%, and 13.3-21.7%, respectively, as a result of 0.2-0.8% addition of HZ. Significant positive linear correlations were found between heavy metal concentrations in TCLP extracts and those in rice tissues except for Pb. Extracting heavy metals with TCLP was a more suitable method for estimating heavy metal bioavailability concentrations of amended soil than exchangeable fraction of heavy metals, because the latter underestimated heavy metal bioavailability. These results demonstrate that LS and HZ could be effective in reducing heavy metal bioavailability and accumulation in rice grown on multi-metal-contaminated soils., (Copyright © 2014 Elsevier Inc. All rights reserved.)

A pot experiment was conducted to study the effects of a group matching curing agent (limestone plus sepiolite) with different concentrations (0, 1.0, 2.0, 4.0, 8.0, 16.0 g x kg(-1)) on the exchangeable Pb, Cd, and Zn contents in the potted soil collected from a mining area in southern China and their cumulative distribution in rice plants. The results showed that: (1) Exchangeable Pb, Cd, and Zn contents in the potted soils decreased obviously with the increase in the amounts of group matching curing agent by 15.3%-99.9%, 9.2%-99.9%, and 7.0%-99.9%, respectively, compared with the CK, indicating that the addition of limestone plus sepiolite effectively inhibited the uptake of Pb, Cd, Zn from soils into rice plants. (2) Growth conditions of rice plants were improved, and the heights of rice plants increased with the increase in the amounts of group matching curing agent. The rice yield reached the optimal level and was increased by 60.4% compared with the CK, when the amount of limestone plus sepiolite was 1.0 g x kg(-1). The uptake of heavy metals in different organs of rice plants decreased obviously with the increasing amounts of the group matching curing agent, and the Pb, Cd, and Zn contents of rice grains decreased by 2.2%-13.1%, 29.3%-79.3%, 19.5%-43.3% with different concentrations (1.0-16.0 g x kg(-1)) of the group matching curing agent, respectively, compared with the CK.

The goal of this study was to determine whether ultrasound (US) exposure combined with microbubble destruction could be used to enhance non-viral gene delivery in rat C6 glioma cells. Microbubbles were prepared and gently mixed with plasmid DNA. The mixture of the DNA and microbubbles was administered to cultured C6 cells under different US/microbubble conditions. Transfection efficiency and cell viability were assessed by FACS analysis, confocal laser scanning microscopy, and Trypan blue staining. The results demonstrate that microbubble with US exposure could significantly enhance the reporter gene expression as compared with other groups. No statistical significant difference was observed in the glioma cell viability between different groups. Our in vitro findings suggest that US-mediated microbubble destruction has the potential to promote safe and efficient gene transfer into C6 cells. This non-invasive gene transfer method may be useful for safe clinical gene therapy of brain cancer without a viral vector system.

Objectives: Previously, we have found that the ClC-3 chloride channel is involved in endothelin-1 (ET-1)-induced rat aortic smooth muscle cell proliferation. The present study was to investigate the role of ClC-3 in cell cycle progression/distribution and the underlying mechanisms of proliferation., Materials and Methods: Small interference RNA (siRNA) is used to silence ClC-3 expression. Cell proliferation, cell cycle distribution and protein expression were measured or detected with cell counting, bromodeoxyuridine (BrdU) incorporation, Western blot and flow cytometric assays respectively., Results: ET-1-induced rat basilar vascular smooth muscle cell (BASMC) proliferation was parallel to a significant increase in endogenous expression of ClC-3 protein. Silence of ClC-3 by siRNA inhibited expression of ClC-3 protein, prevented an increase in BrdU incorporation and cell number induced by ET-1. Silence of ClC-3 also caused cell cycle arrest in G(0)/G(1) phase and prevented the cells' progression from G(1) to S phase. Knockdown of ClC-3 potently inhibited cyclin D1 and cyclin E expression and increased cyclin-dependent kinase inhibitors (CDKIs) p27(KIP) and p21(CIP) expression. Furthermore, ClC-3 knockdown significantly attenuated phosphorylation of Akt and glycogen synthase kinase-3beta (GSK-3beta) induced by ET-1., Conclusion: Silence of ClC-3 protein effectively suppressed phosphorylation of the Akt/GSK-3beta signal pathway, resulting in down-regulation of cyclin D1 and cyclin E, and up-regulation of p27(KIP) and p21(CIP). In these BASMCs, integrated effects lead to cell cycle G(1)/S arrest and inhibition of cell proliferation.

Aim: To examine how the sarcoplasmic reticulum (SR) Ca2+ content changes and the underlying mechanism in L-thyroxin-induced cardiac hypertrophy., Methods: Echocardiography was used to confirm the establishment of the cardiac hypertrophy model. The confocal microscopy and fluorescent indicator Fluo-3 was applied to examine the intracellular Ca2+ concentration ([Ca2+]i), the Ca2+ sparks, and the caffeine-induced Ca2+ transient in freshly isolated cardiac ventricular myocytes. The activity of sarcolemmal and SR Ca2+-ATPase 2a (SERCA2a) in the ventricular tissue was also measured, respectively., Results: L-thyroxin (1 mg/kg injection for 10 d) induces left ventricular cardiac hypertrophy with normal myocardial function. The decreased caffeine-induced Ca2+ transient in the Ca2+-free solution was detected. The spontaneous Ca2+ sparks in hypertrophied myocytes occurred more frequently than in normal cells, with similar duration and spatial spread, but smaller amplitude. Then the basal [Ca2+]i increase was observed in quiescent left ventricular myocytes from hyperthyroidism rats. The activity of sarcolemmal and SR Ca2+-ATPase was decreased in the hypertrophied ventricle tissue., Conclusion: The results suggested that the reduced SR Ca2+ content may be associated with an increased Ca2+ leak and reduced SERCA2a activity, contributing to abnormal intracellular Ca2+ handling during hypertrophy in hyperthyroidism rats.

Objective: To observe the effects of ginsenoside (Gs) and berberine (Ber), two kinds of active components of traditional Chinese herbal medicine, on transforming growth factor-beta1 (TGF-beta1) and prostaglandin E2 (PGE2) in PG cells., Methods: Co-culture system of human lung carcinoma cell line PG and human T lymphocyte cell line Jurkat was established. PG cells were treated with Gs (100 microg/ml) and Ber (10 mug/ml) for twenty-four hours, and then cocultured with Jurkat cells. After 24-hour coculture, the state of Jurkat cells was observed with inverted microscope. The viable count of Jurkat cells was detected by trypan blue staining after 6- and 24-hour coculture, and the apoptosis of Jurkat cells was evaluated by flow cytometry. PG cells were treated with 100, 50, 25 microg/ml Gs and 10, 5, 2.5 microg/ml Ber respectively, and the content of TGF-beta1 and PGE(2) in PG cells was detected by enzyme-linked immunosorbent assay (ELISA) and radioimmunoassay (RIA) method., Results: After coculture with PG cells treated with Gs and Ber, the number of Jurkat cells was less than blank control group, and the apoptosis rates of Jurkat cells in Gs- and Ber-treated groups were higher than blank control group. Gs and Ber could promote the secretion of TGF-beta1 in PG cells, but could not change the level of PGE(2)., Conclusion: Gs and Ber can promote the growth inhibition and apoptosis of Jurkat cells induced by PG cells, which may be related to the up-regulation of Gs and Ber on TGF-beta1 secretion in PG cells.

The cerebrovascular remodeling is a prominent feature of hypertension and considered a major risk factor for stroke. Cerebrovascular smooth muscle cells meet volume challenge during this pathophysiological process. Our previous studies suggest that volume regulated chloride channels may be critical to the cell cycle of vascular smooth muscle cells. However, it is unknown whether the volume-regulated chloride movement is altered in hypertension. Therefore, we directly measured the concentration of intracellular chloride ([Cl(-)](i)) in rat basilar arterial smooth muscle cells isolated from control rats and rats that were made hypertensive for 1 to 12 weeks after partial renal artery constriction (2-kidney, 2-clip method) using a 6-methoxy-N-ethylquinolinium iodide fluorescence probe. The [Cl(-)](i) in isotonic solution showed no difference in all of the groups. After hypotonic perfusion, the reduction in [Cl(-)](i) was more prominent in hypertensive cerebrovascular smooth muscle cells than in sham control cells. Genistein, a protein tyrosine kinase inhibitor, inhibited hypotonic-induced reduction in [Cl(-)](i), whereas sodium orthovanadate, a protein-tyrosine phosphatase inhibitor, enhanced hypotonic-induced reduction in [Cl(-)](i) in both groups. The percentage inhibition of reduction in [Cl(-)](i) by genistein on volume-regulated chloride movement has a positive correlation with blood pressure levels in the 2-kidney, 2-clip hypertensive group, as is the case for the percentage increase of reduction in [Cl(-)](i) by sodium orthovanadate. Antihypertensive therapy with the angiotensin-converting enzyme inhibitor captopril completely reversed abnormal volume-regulated chloride movement in hypertensive rats. We conclude that volume-regulated chloride movement is augmented in rat cerebrovascular smooth muscle cells in proportion to the severity of hypertension.

In normal condition, 150 Fourier transform infrared-attenuated total reflection spectra of three kinds of tobacco flavor were measured. Mean centering and Karl Norris second derivative filter were employed for treating the spectra data. Whereafter, class models of tobacco flavor were established with classification of principal component analysis-Mahalanobis distance. Class models have been applied to multivariate statistical process control(MSPC) for the quality assessment of tobacco flavor in site. The upper control limit(UCL) was based on this kind of average Mahalanobis distance within +3sigma. The results of monitoring are correct.

Previously, it was found that total saponins from panax notoginseng inhibited Ca2+ influx coupling to activation of alpha1-adrenoceptor. This study was designed to investigate the effects of ginsenoside-Rd from total saponins of panax notoginseng on receptor-operated (ROCC) and store-operated (SOCC) Ca2+ channels in vascular smooth muscle cells using fura-2 fluorescence, whole cell patch clamp ion channel recording, radio-ligand-receptor binding, 45Ca2+ radio-trace and organ bath techniques. It was found that ginsenoside-Rd reduced phenylephrine-induced contractile responses and Ca2+ influx in normal media without significant effect on these responses in Ca2+ -free media. Ginsenoside-Rd also decreased phenylephrine- and thapsigargin-induced inward Ca2+ currents, and attenuated thapsigargin- and 1-oleoy-2-acetyl-sn-glycerol (OAG)-induced cation entries that are coupled to ROCC and SOCC respectively. Ginsenoside-Rd failed to inhibit KCl-induced contraction of rat aortal rings and Ca2+ influx, and did not alter voltage-dependent inward Ca2+ current (VDCC) which was blocked by nifedipine. Also, ginsenoside-Rd did not change binding site and affinity of [3H]-prazosin for alpha1-adrenoceptor in the vascular plasma membrane. These results suggest that ginsenoside-Rd, as an inhibitor, remarkably inhibits Ca2+ entry through ROCC and SOCC without effects on VDCC and Ca2+ release in vascular smooth muscle cells.

Aim: To test the hypothesis that Cl channel blockers affect T cell proliferation through Ca2+-release-activated Ca2+ (CRAC) signaling and examine the effects of the combination of a CRAC channel blocker and a Cl channel blocker on concanavalin A (ConA; 5 mg/mL)-induced Ca2+ signaling, gene expression and cellular proliferation in human peripheral T lymphocytes., Methods: [3H]Thymidine incorporation, Fura-2 fluorescent probe, RNase protection assay, and reverse transcription-polymerase chain reaction were used., Results: The Cl channel blocker 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS) inhibited ConA-induced Ca2+ influx, interleukin-2 mRNA expression and T lymphocyte proliferation in a concentration-dependent manner, and also enhanced the inhibitory effects of 1-[beta-[3-(4-methoxyphenyl)propoxyl]-4-methoxyphenethyl]-1H-imidazole (SK&F96365) on the above key events during T cell activation. A combination of DIDS (1 micromol/L) and SK&F96365 (1 micromol/L) significantly diminished ConA-induced ClC-3 mRNA expression by 64%, whereas DIDS(1 micromol/L) or SK&F96365 (1 micromol/L) alone decreased ConA-induced ClC-3 mRNA expression by only 16% and 9%, respectively., Conclusion: These results suggest that there is an interaction between CRAC-mediated Ca2+ signaling and DIDS-sensitive Cl channels during ConA-induced T cell activation and proliferation. Moreover, the DIDS-sensitive Cl channels may be related to the ClC-3 Cl channels.

Cell volume can be altered by two different ways, swelling and shrinkage. Cell swelling is regulated by volume-regulated Cl- channel (VRC). It is not well understood whether shrinkage is regulated by VRC. We previously found that antisense oligonucleotide specific to ClC-3 (ClC-3 antisense) prevented cell proliferation, which was related to cell swell volume regulation. In the present study, we further studied the role of ClC-3 Cl- channel in cell apoptosis which was related to cell shrinkage volume regulation by using antisense oligonucleotide specific to ClC-3 (ClC-3 antisense) and ClC-3 cDNA transfection techniques. We found that thapsigargin (TG), a specific inhibitor of the endoplasmic reticulum calcium ATPase, evoked apoptotic morphological changes (including cytoplasmic blebbing, condensation of nuclear chromatin, and the formation of apoptotic bodies), DNA laddering, and caspase-3 activation in PC12 cells (Pheochromocytoma-derived cell line). TG increased the cell apoptotic population with a decrease in cell viability. These effects were consistent with the decrease in endogenous ClC-3 protein expression, which was also induced by TG. Overexpression of ClC-3 significantly inhibited TG effect on PC12 cell apoptosis, whereas the ClC-3 antisense produced opposite effects and facilitated apoptosis induced by TG. Our data strongly suggest that ClC-3 channel in PC12 cells mediates TG-induced apoptotic process through inhibitory mechanism. Thus, it appears that ClC-3 Cl- channel mediates both cell proliferation and apoptosis through accelerative and inhibitory fashions, respectively.

It is generally thought that receptor-operated Ca2+ entry is related to store-operated or capacitative Ca2+ entry mechanism. Recent evidence suggests that non-capacitative Ca2+ entry pathways are also involved in receptor activated Ca2+ influx in many different kinds of cells. In this study, we studied whether alpha1-adrenoreceptor (alpha1-AR)-activated Ca2+ entry is coupled to both capacitative and non-capacitative pathways in A10 vascular smooth muscle cells by fura-2 fluorescence probe and conventional whole-cell patch clamp techniques. We found that both thapsigargin (TG) and phenylephrine (Phe) induced transient increase in cytoplasmic Ca2+ concentration ([Ca2+]i) in Ca2+-free medium, and subsequent addition of Ca2+ evoked a sustained [Ca2+]i rise. When the membrane potential was held at -60 mV, both TG and Phe activated inward currents, which were inhibited by GdCl3(Gd3+), 0Na+/0Ca2+ solution and 1-{beta[3-(4-mehtoxyphenyl)propoxy]-4-methoxypheneth-yl}-1H- imidazole hydro-chloride (SK&F96365), but not by nifedipine. When Ca2+ store was depleted by TG in Ca2+-free solution, Phe failed to further evoke [Ca2+]i rise. However, when capacitative Ca2+ entry was activated by TG in the medium containing Ca2+, 10 microM Phe further increased [Ca2+]i. At the same concentration, TG activated an inward cation current, subsequent addition of Phe also further induced an inward cation current. Furthermore, the amplitudes of [Ca2+]i increase and current density induced by Phe in the presence of TG were less than that induced by Phe alone. Our results suggest that both capacitative and non-capacitative Ca2+ entry pathways are involved in Ca2+ influx induced by activation of alpha1-AR in A10 vascular smooth muscle cells.

Objective: To evaluate the effect of health education on lung function and quality of life in stabilized patients with chronic pulmonary disease (COPD)., Methods: 117 stabilized COPD patients were randomly devided into 4 groups with numbers as 31,26, 20 and 40 identified as Groups 1 to 4. Patients in Group 1 did not receive health education, but Groups 2,3 and 4 received one, two, three or more times health education in file. FEV1, FEV1%, FEV1/FVC and SGRQ score were compared pre and 6-month post the health education program., Results: Health education seemed successful in delaying the decline of FEV1, FEV%, FEV1/FVC and groups 2-4 were superior to group 1(P < 0.05) while groups 3 and 4 were superior to groups 1 or 2(P < 0.05). Health education was effective in raising the SGRQ score among the stabilized COPD patients with groups 2-4 superior to group 1 (P < 0.05) while groups 3 and 4 superior to groups 1 or 2 (P < 0.05)., Conclusion: Health education could effectively delay the decline of both lung function and quality of life in stabilized patients with COPD.

Objective: To study the therapeutic effects and mechanism of Jiechangning (JCN) decoction on carrageenan induced experimental ulcerative colitis (UC)., Methods: After sensitizing guinea pigs with carrageenan, we established UC animal models by free drinking water containing 2% acid degraded carrageenan (ADC). JCN decoction was orally administered once a day for 2 weeks after carrageenan treatment. Salicylazosulfapyridine (SASP) and normal saline were given to the other two groups as control. The levels of colon lipid peroxide (LPO), acid phosphatase (ACP) activity and tumor necrosis factor-alpha (TNF-alpha) were measured; colitis activity score (CAS) was carried out for assessment of the degree of tissue inflammation and injury; the colonic pathological changes were examined simultaneously with hematoxylin and eosin (HE) and toluidine blue staining used to evaluate the therapeutic effects of JCN decoction and SASP., Results: Experimental colitis models resembling human UC were successfully induced. The levels of tissue LPO, ACP activity and the content of tissue TNF-alpha were markedly increased in the model group as compared with the normal control group (P < 0.01) and were positively correlated with CAS. JCN decoction could reverse these changes like SASP. HE staining showed that JCN decoction and SASP could reduce CAS and the degree of tissue injury, toluidine blue staining revealed that mucosa and submucosa red metachromasia pellets in JCN group and SASP group were markedly fewer than those in the model group., Conclusion: JCN decoction is effective in treating experimental UC, which provides theoretical basis for its clinical application.

We previously found that antisense oligonucleotide specific to ClC-3 (ClC-3 antisense) prevented rat aortic smooth muscle cell proliferation, which was related to cell volume regulation. In the present study, we further characterized the regulation of intracellular Cl(-) concentrations ([Cl(-)](i)) via volume-regulated ClC-3 Cl(-) channels in an embryo rat aortic vascular smooth muscle cell line (A10 cell) and ClC-3 cDNA-transfected A10 cells (ClC-3-A10) using multiple approaches including [Cl(-)](i) measurement, whole cell patch clamp, and application of ClC-3 antisense and intracellular dialysis of an anti-ClC-3 antibody. We found that hypotonic solution decreased [Cl(-)](i) and evoked a native I(Cl.vol) in A10 cells. The responses of [Cl(-)](i) and I(Cl.vol) to hypotonic challenge were enhanced by expression of ClC-3, and inhibited by ClC-3 antisense. The currents in A10 (I(Cl.vol)) and in ClC-3-A10 cells (I(Cl.ClC-3)) were remarkably inhibited by intracellular dialysis of anti-ClC-3 antibody. Reduction in [Cl(-)](i) and activation of I(Cl.vol) and I(Cl.ClC-3) in A10 and ClC-3-A10 cells, respectively, were significantly inhibited by activation of protein kinase C (PKC) by phorbol-12,13-dibutyrate (PDBu) and inhibition of tyrosine protein kinase by genistein. Sodium orthovanadate (vanadate), a protein-tyrosine phosphatase inhibitor, however, enhanced the cell swelling-induced reduction in [Cl(-)](i), accompanied by the activation of I(Cl.vol) and I(Cl.ClC-3) in a voltage-independent manner. Our results suggest that the volume-regulated ClC-3 Cl(-) channels play important role in the regulation of [Cl(-)](i) and cell proliferation of vascular smooth muscle cells.

Objective: To explore the detoxication and tissue generation effect of Astragalus (As) in wound healing and its relation with inflammatory reaction, through observing the effect of Astragalus polysaccharides (AP) on neutrophil-endothelial cell adhesion and expression of related adhesive molecules., Methods: Human polymorphonuclear leucocyte (PMN) or human umbilical vein endothelial cell (HUVEC) was treated separately with AP, AP plus interleukin 1 (IL-1) and tumor necrosis factor (TNF) to study the effect of AP on PMN adhesion with HUVEC by rose bengal staining, and that on expression of superficial adhesive factor by means of Cell-ELISA and APAAP method., Results: When AP acted on HUVEC, it could significantly promote the adhesion of HUVEC with PMN, while when AP acted on PMN, the adhesion would not increase. When HUVEC was treated by AP plus IL-1, the IL-1 induced PMN adhesion with HUVEC could be strengthened, and the expression of HUVEC superficial adhesive factor ICAM-1 induced by IL-1 and TNF was strengthened also, but when PMN treated with AP, it showed no effect on the expression of adhesive factor CD18., Conclusion: AP promotes the adhesion between neutrophil and endothelial cell by way of promoting the expression of superficial I-CAM-1 on surface of endothelial cells, so as to improve the inflammatory reaction in the wound healing course, it possibly is one of the biological bases of the detoxication and tissue generation effects of AP.

The therapeutic effect of arsenic trioxide (As2O3) in the treatment of acute promyelocytic leukemia (APL) was evaluated among 15 APL patients at relapse after all-trans retinoic acid (ATRA) induced and chemotherapy maintained complete remission (CR). As2O3 was administered intravenously at the dose of 10 mg/d. Clinical CR was achieved in nine of 10 (90%) patients treated with As2O3 alone and in the remaining five patients treated by the combination of As2O3 and low-dose chemotherapeutic drugs or ATRA. During the treatment with As2O3, there was no bone marrow depression and only limited side effects were encountered. Pharmacokinetic studies, which were performed in eight patients, showed that after a peak level of 5.54 micromol/L to 7.30 micromol/L, plasma arsenic was rapidly eliminated, and the continuous administration of As2O3 did not alter its pharmacokinetic behaviors. In addition, increased amounts of arsenic appeared in the urine, with a daily excretion accounting for approximately 1% to 8% of the total daily dose administered. Arsenic contents in hair and nail were increased, and the peak content of arsenic could reach 2.5 to 2.7 microg/g tissue at CR. On the other hand, a decline of the arsenic content in hair and nail was observed after withdrawal of the drug. We conclude that As2O3 treatment is an effective and relatively safe drug in APL patients refractory to ATRA and conventional chemotherapy.

Molecular cloning of several homing receptors and their placement within unique families of adhesion receptors over the past 2 years will now permit detailed analyses of structure, function and regulation. Novel tools have significantly contributed to the characterization of carbohydrates as essential parts of the recognition site in addressins whose molecular structures remain to be elucidated.