Prognostic utility of 99mTc-MIBI single photon emission computerized tomography myocardial perfusion imaging in patients with ischemia and non-obstructive coronary artery disease.

Objective: The aim of our study was to evaluate the prognostic value of gated SPECT MPI in non-obstructed coronary arteries (INOCA) patients, sought to stratify patients more accurately and thus derive more reliable prognostic information. Materials and methods: In total, 167 patients with INOCA were enrolled. The patients were divided into two groups according to their SSS. Patients were followed-up regularly in terms of major adverse cardiovascular event (MACE), including cardiac death, nonfatal myocardial infarction, stroke, re-hospitalization with angina pectoris, and recurrent angina pectoris. Kaplan-Meier curves and Cox's proportional hazards models were used to analyze survival and identify predictive factors. Results: Adverse cardiac events occurred in 33 cases (19.8%). The rate of MACE was higher in the summed stress score (SSS) ≥4 group than in the SSS 0-3 group (30.1% vs. 9.5%, respectively, P = 0.001) and MACE-free survival was lower (annual MACE-free rates of 87.5% vs. 96.2%, respectively, P = 0.003). Event-free survival was consistently higher in patients with normal arteries than in those with non-obstructive coronary artery disease (annual MACE-free rates of 96.1% and 88.4%, P = 0.035). When the SSS and the CAG results were combined, patients with normal coronary arteries (SSS 0-3) had the best prognosis and those with non-obstructive coronary artery stenosis (SSS ≥ 4) had the worst. However, the early prognosis of patients with non-obstructive coronary artery disease and SSS of 0-3 was comparable to that of patients with normal coronary arteries and SSS ≥ 4 (annual MACE-free rates of 100%, 94.6%, 93.1%, and 78.2%, respectively). Multivariate Cox's regression indicated that the SSS [hazard ratio (HR) = 1.126, 95% confidence interval (CI) 1.042-1.217, P = 0.003] and non-obstructive coronary artery disease (HR = 2.559, 95% CI 1.249-5.246, P = 0.01) were predictors of adverse cardiac events. Conclusion: SPECT MPI data were prognostic for INOCA patients, thus identifying groups at high risk. The long-term predictive efficacy of such data exceeded that of CAG data. A combination of the two measures more accurately stratified INOCA patients in terms of risk.

Chlorin e6-1,3-diphenylisobenzofuran polymer hybrid nanoparticles for singlet oxygen-detection photodynamic abaltion.

A dual-functional nanosysterm is developed by means of Chlorin e6 (Ce6) as photosensitizer and 1,3-Diphenylisobenzofuran (DPBF) as fluorescent singlet oxygen (1O2) probe. Under 660 nm laser irradiation, Ce6 exhibites efficient 1O2 generation, and subsequently the production of 1O2 is assessed by the ratiometric fluorescence of PFO and DPBF under one-photon and two-photon excitation mode. The nanoparticles with excellent biocompatibility can be internalized into Hela cells and applied for tumor treatment. For intracellular PDT, the nanoparticles perform a high phototoxicity, while the PDT proccess can be evaluated in time by monitoring fluorescence signals of DPBF. This theranostic nanosysterm provides a facile strategy to fabricate 1O2-detection PDT, which can realize accurate and efficient photodynamic therapy based on singlet oxygen detection.

Facile synthesis of polypyrrole-rhodamine B nanoparticles for self-monitored photothermal therapy of cancer cells.

Photothermal therapy following microscopic temperature detection can avoid overheating effects or insufficient heating, and thus improve therapeutic efficacy. In this study, biocompatible dual-functional nanoparticles (NPs) are constructed from polypyrrole (PPy) and rhodamine B (RB) by a one-step modified polymerization method. The polypyrrole serves as a photothemal agent, and rhodamine B acts as a temperature-sensing probe. The polypyrrole-rhodamine B (PPy-RB) NPs possess a high photothermal effect on irradiation by 808 nm laser, and a competent temperature sensitivity for the real-time temperature monitoring based on the emission intensity response of rhodamine B. After acting on HepG2 cells, the PPy-RB NPs can effectively induce cancer cell death, and the microscopic temperature is monitored by fluorescence feedback from rhodamine B during PTT by laser confocal microscopy. Hence, the proposed approach can supply a facile and promising way for the fabrication of effective theranostic nanoplatforms assisted by self-monitoring of cancer therapeutic processes.

TGF-ß inhibition via CRISPR promotes the long-term efficacy of CAR T cells against solid tumors.

In recent years, chimeric antigen receptor-modified T cell (CAR T cell) therapy has proven to be a promising approach against cancer. Nonetheless, this approach still faces multiple challenges in eliminating solid tumors, one of which being the immunosuppressive tumor microenvironment (TME). Here, we demonstrated that knocking out the endogenous TGF-ß receptor II (TGFBR2) in CAR T cells with CRISPR/Cas9 technology could reduce the induced Treg conversion and prevent the exhaustion of CAR T ce lls. Meanwhile, TGFBR2-edited CAR T cells had better in vivo tumor elimination efficacy, both in cell line-derived xenograft and patient-derived xenograft solid tumor models, whether administered locally or systemically. In addition, the TGFBR2-edited CAR T cells could eliminate contralaterally reinoculated xenografts in mice effectively, with an increased proportion of memory subsets within circulating CAR T cells of central memory and effector memory subsets. In conclusion, we greatly improved the in vitro and in vivo function of CAR T cells in TGF-ß-rich tumor environments by knocking out endogenous TGFBR2 and propose a potentially new method to improve the efficacy of CAR T cell therapy for treating solid tumors.

N-ethylmaleimide­sensitive factor siRNA inhibits the release of Weibel-Palade bodies in endothelial cells.

The aim of the present study was to examine the effect of small interfering RNA (siRNA) methods on the expression of N­ethylmaleimide sensitive factor (NSF) and Weibel­Palade body (WPB) release in endothelial cells. A small hairpin RNA (shRNA), mediated with an adenovirus vector, was designed to target the N­terminal functional area of NSF. Subsequently, viruses were transfected into human aortic endothelial cells. The mRNA and protein expression levels of NSF were detected using reverse transcription­quantitative polymerase chain reaction and Western blot analyses, respectively, and the release of WPBs in the endothelial cells was examined using immunofluorescence. The mRNA expression of NSF in the endothelial cells, which were transfected with the adenoviruses carrying the NSF­shRNA was significantly decreased, compared with the negative control group (P=0.035) and blank control group (P=0.02). In addition, the mRNA expression of NSF was gradually decreased as duration increased; there were marked differences between the 24, 48 and 72 h groups (P<0.05). The protein expression of NSF was significantly decreased in the experimental group, compared with the negative control group (P=0.004) and blank control group (P=0.031), however, no difference was observed between the negative control and blank control groups (P=0.249). The immunofluorescence staining showed that the release of WPBs in the endothelial cells induced with thrombin was inhibited markedly following transfection with the virus carrying the NSF­shRNA. Therefore NSF­siRNA inhibited the mRNA and protein expression levels of NSF, and inhibited the release of WPBs in endothelial cells induced with thrombin. These results suggested that NSF-siRNA may be valuable for preventing and treating atherosclerosis and acute coronary syndrome.

[Characteristics and Functional Status of Victims with Fractured Injury in Lushan Earthquake].

OBJECTIVE: To analyze the functional limitations and characteristics of victims with fractured injury in Lushan Earthquake. METHODS: A total of 113 patients admitted to the hospital from April 20th to 30th 2013 for treatment of fractured injury in Lushan Earthquake were recruited for this study. They were evaluated by the department of rehabilitation medicine in West China Hospital using a questionnaire. RESULTS: The most common cause of injury was falling (39.82%). Vertebral fractures (26.55%) was the most common type of injury. Most injure was light (80.53%), but incidence of complications was high (89.37%). Motor functional limitations included restricted range of motion (97.35%), balance functional limitation (89.38%) and decreased muscle force (87.61%). Limited cardiopulmonary function and limited circular function occurred in about 19.47% of patients. About 76. 11% patients had restricted activities of daily living capacities, predominantly in bathing (96.46%), stair climbing (86.73%), and walking (78.76%). Pain symptoms were common (97.35%), with 4-6 in visual analogue scale (VAS) values accounting for 40.71% of those reporting pains. Limited psychological function, professional function and social function was reported by 46.02%, 99.12% and 100% patients, respectively. CONCLUSION: Fractured injury of Lushan earthquake victims has its own characteristics. Increased attention should be paid to the restoration of psychological function, professional function and social function.

[Effects of whole body vibration on structural and functional remodeling of subchondral bones in rabbits with early osteoarthritis].

OBJECTIVE: To investigate the effects of whole body vibration therapy (WBVT) on structural and functional remodeling of subchondral bone in rabbits with early Osteoarthritis (OA). METHODS: Twenty-four adult rabbits received anterior cruciate ligament transaction (ACLT) to establish knee osteoarthritis model. The rabbits were randomly divided into two groups: ACLT group and WBVT+ ACLT group. The rabbits in the WBVT+ ACLT group received whole body vibration treatment (Frequency: 40 Hz; Amplitude: 2-4 mm; 40 min/d, 5 d/week, 4 weeks) two months after surgery. After interventions, the left femur and tibia of all rabbits were placed in a Micro-CT scanning system. The bone volume fraction (BVF), trabecular thickness (Tb. Th), trabecular spacing (Tb. Sp), trabecular number (Tb. N), volumetric BMD (vBMD), and tissue BMD (tBMD) of the bones of femoral condyle and tibia were analysed using three-dimensional reconstruction Micview V2.1.2 and ABA analysis software. Geomagic Studio 11.0 software were used to calculate modulus (EM), reaction force (RF) and average Von Miss stress (VMF). RESULTS: Higher levels of BVF, Tb. N, Tb. Th, EM, RF, VMF, vBMD and tBMD and lower levels of Tb. Sp were found in distal femora and tibia of the rabbits in the WBVT+ACLT group compared with the controls (P < 0.05). CONCLUSION: WBVT can effectively improve bone microstructure and mechanical properties of rabbits with early knee osteoarthritis.

[Effect of extracorporeal shock wave therapy on cartilage and subchondral bone remodeling in rabbits with ACLT-induced osteoarthritis].

OBJECTIVE: To investigate the role of extracorporeal shock wave therapy on the protection of cartilage and subchondral bone remodeling in rabbits with osteoarthritis induced by anterior cruciate ligament transaction (ACLT). METHODS: Anterior cruciate ligament transaction was performed on 24 adult rabbits to establish knee osteoarthritis models. The rabbits were randomly divided into two groups, with one group receiving extracorporeal shock wave therapy (ESWT) and the other group serving as controls. The ESWT was set at energy 0.16 MPa/time, frequency 5 Hz, 1 200 shock per side, 3 times per week, and a total of 6 times in 4 weeks. Histological observations were undertaken with articular cartilages under mascroscope and microscope; bone mineral density (BMD) was measured. The subchondral bones of femoralcondyle and tibial plateau were given a bone histomorphometry analysis. The levels of interleukin-1 beta (IL-1beta), tumor necrosis factor alpha (TNF-alpha) and nitric oxide (NO) were detected by ELISA. The expressions of matrix metalloproteinases (MMP-1, MMP 3, MMP-13) and tissue inhibitor of metalloproteinases 1 (TIMP-1) in cartilage were determined by RT-PCR. RESULTS: Lower levels of histological result in medial femoral condyles (by both mascroscope and microscope examinations) and higher levels of BMD (in distal femora) were found in the ESWT treated rabbits compared with the controls (P < 0.05). The ESWT treated rabbits also had decreased trabecular bone relative volume and trabecular bone thickness, increased bone separation, lower levels of expressions of MMP-1 and MMP-3, and higher levels of TIMP-1. CONCLUSION: ESWT can protect cartilage from damages and prevent subchondral sclerosis through regulating MMP-1, MMP-3 and TIMP-1 in cartilages and modulating subchondral bone metabolisms.

CCK-8S increases the firing frequency of CCK-positive neurons and facilitates excitatory synaptic transmission in primary rat hippocampal neurons.

The neuropeptide cholecystokinin octapeptide (CCK) is involved in a variety of brain functions. In the hippocampus, most CCK is released from CCK-positive (CCK+) neurons, but the effects of CCK on CCK+ neurons are poorly understood. We employed primary hippocampal cultures to explore the modulatory effect of CCK on CCK+ neurons. CCK-8S (0.2 µM) was added to the culture medium from day in vitro 2 (DIV-2) to DIV-11. An adenovirus integrated with the CCK promoter was used to label CCK+ neurons. Whole-cell patch clamp recording was carried on to record the electrophysiology properties. The results show that: (1) CCK-8S significantly decreased membrane capacity but increased the membrane resistance (Rm) of CCK+ neurons, (2) CCK-8S increased action potential (AP) firing frequency of CCK+ neurons but did not affect the firing pattern, (3) CCK-8S facilitated CCK+ neuron excitatory synaptic transmission but attenuated inhibitory synaptic transmission, and (4) the expression of postsynaptic density-95 (PSD-95) in cultured hippocampal neurons was elevated by CCK-8S treatment. Our results demonstrate that CCK-8S significantly alters the membrane electrophysiological characteristics and synaptic activity of cultured hippocampal CCK+ neurons. These findings may enhance our understanding of the modulatory effect of CCK in the brain.

CCK-8S increased the filopodia and spines density in cultured hippocampal neurons of APP/PS1 and wild-type mice.

Cholecystokinin (CCK), a neuropeptide, is widely distributed in the brain. The function of CCK is involved in many brain functions including learning and memory, but the cellular mechanism is poorly understood. In the present study, we investigated the effect of CCK on dendritic filopodia and spines of cultured hippocampal neurons from wild-type and APP/PS1 mice. The cultured hippocampal neurons were infected with CMV-GFP (CMV promoter with green fluorescent protein) adenovirus 24h before image acquisition to display the subtle structure of dendrites. Cholecystokinin octapeptide sulfated (CCK-8S, 0.2µM) was added into the cultured solution from divided in vitro day 2 (DIV 2). A decrease of filopodia and spines density was observed in APP/PS1 mice compared with that of wild type mice. CCK-8S increased the density of filopodia and spines at DIV 7, DIV 14 and DIV 21 in hippocampal neurons of both wild-type and APP/PS1 mice. In addition, this effect was inhibited by CI988, an antagonist of CCK-2 receptor. Those results indicate that CCK-8S can influence the dendritic development and spine genesis of cultured hippocampal neurons derived from both wild-type and APP/PS1 mice. These data suggest that CCK may play an important role in learning and memory.

[Leaching experiments on the release of trace elements from tailings of Chashan antimony mine, Guangxi, China].

The leaching of trace elements from tailings of an antimony mine in Guangxi Autonomous Region, China, was investigated through column leaching under wet-dry cycling and complete immersion conditions. Simulated acid rain (pH 4.0-4.4) and river water (pH 8.0) were used as the leaching solution. No matter the simulated acid rain or river water was used, the leachate always showed a slightly alkaline pH between 7.2 and 8.0, suggesting an acid neutralization capacity of the tailing. Compared to As and Pb, Sb was leached out to a much higher extent in this circumstance. Furthermore, Sb release was largely enhanced in wet-dry cycle compared to the complete immersion condition. In contrast, As was leached more readily in the complete immersion condition, and the longer the tailings were immersed in water, the higher the As concentration in the leachate. The leachate on day 5 and day 10 showed 1-2 times higher As concentration as compared with the leachate on day 1 and day 2. The leaching of Mn and Zn by simulated acid rain was much stronger than that by river water, and the release of Mn and Zn was more significantly affected by pH than by O2 (i.e., the difference between the wet-dry cycle and complete immersion condition). Sr showed a high release rate that was not affected by leaching solution or air-exposure condition. Basically, Pb showed a very low leaching potential. In general, an alkaline circumstance combined with wet-dry cycle forms the favorable condition for the release of Sb in the tailings.

[Characteristics of heavy metals in soil profile and pore water around Hechi antimony-lead smelter, Guangxi, China].

In the area around an antimony-lead smelter, soil and soil water samples were collected from different soil profiles in vegetable field, uncultivated field and wood field. The distribution and migration of heavy metals in soil profile were investigated. In soil, Sb, Pb, Cu and Cd are found to accumulate in the topsoil layer, and decrease rapidly with the depth. However, Sb, Pb, and As appear to accumulate below 60 cm. Sb and As show a much stronger affinity to the pore water than Pb. In consequence, different soil utilization types lead to different effects of heavy metals migration in soil profile. Furthermore, Sb and As show a higher potential to migrate along the soil profile than that of Pb. In addition, the high content of heavy metals in the topsoil may be absorbed by crops post to desorption in soil water, and possibly form a threat to human health.

The effect of ultrasound on lipase-catalyzed regioselective acylation of mangiferin in non-aqueous solvents.

A simple and efficient method for regioselective acylation of mangiferin catalyzed by lipase under ultrasound irradiation is reported. Compared with the conventional methods, its main advantages are shorter reaction time and higher yields. The optimum conditions were screened out. Under the optimal conditions (lipase: PCL, acyl donor: vinyl acetate; reaction solvent: DMSO, reaction temperature: 45 degrees C, ultrasonic power: 200 W; substrate ratio: acyl donor/mangiferin 6/1, enzyme loading: 6 mg/ml), the regioselective acylation yield was up to 84%.

Enantioselective transesterification of glycidol catalysed by a novel lipase expressed from Bacillus subtilis.

A novel plasmid (pBSR2) was constructed by incorporating a strong lipase promoter and a terminator into the original pBD64. The lipase gene from Bacillus subtilis strain IFFI10210 was cloned into the plasmid pBSR2 and transformed into B. subtilis A.S.1.1655 to obtain an overexpression strain. The recombinant lipase [BSL2 (B. subtilis lipase 2)] has been expressed from the novel constructed strain and used in kinetic resolution of glycidol through enantioselective transesterification. The effects of reaction conditions on the activity as well as enantioselectivity were investigated. BSL2 showed a satisfying enantioselectivity (E>30) under the optimum conditions [acyl donor: vinyl butyrate; the mole ratio of vinyl butyrate to glycidol was 3:1; organic medium: 1,2-dichloroethane with water activity (a(w))=0.33; temperature 40 degrees C]. The remaining (R)-glycidol with a high enantiomeric purity [ee (enantiomeric excess) >99%] could be obtained when the conversion was approx. 60%. The results clearly show a good potential for industrial application of BSL2 in the resolution of glycidol through enantioselective transesterification.

[Regulatory network of hippocampal-systemic arterial blood pressure during epileptic network reestablishment].

AIM: To investigate the regulatory network of hippocampal-systemic arterial blood pressure during epileptic network reestablishment. METHODS: 7.2 microg picrotoxin (PTX) was microinjected into the right HPC (RHPC) to induce rat epilepsy. Contralateral hippocampal EEG, single unit discharges, femoral artery blood pressure and ECG were recorded simultaneously. RESULTS: PTX might induce: (1) A resemblance interspike intervals (ISI) spot distribution of long duration neuronal burst and unit after discharges in contralateral HPC. (2) Delayed the initiation time of hippocampal neuronal bursts coupled with arterial blood pressure depression. (3) Hippocampal neuronal burst or unit after discharges coupled complexly with arterial blood pressure depression. (4) Resemblance hippocampal EEG interpeak intervals (IPI) and neuronal firing ISI spot distribution coupled with arterial blood pressure depression. CONCLUSION: During contralateral hippocampal epileptic network reestablishment after microinjection of PTX to the RHPC, the function of the hippocampal-arterial blood pressure regulatory network could be modulated by characteristic network and neuronal temporal code patterning.

[Inhibitory effects of human bone marrow mesenchymal stem cells and cord blood mononuclear cells on mixed lymphocyte response and PHA induction transformation].

To study and compare the immunomodulatory functions of human bone marrow mesenchymal stem cell (MSC) and cord blood mononuclear cell (CBMNC) in vitro, human bone marrow MSC were separated with Percoll (1.073 g/L) and cultured in low-glucose DMEM, and cord blood mononuclear cells were isolated with Ficoll (1.077 g/L). These two kinds of cells were added to mixed lymphocyte cultures and PHA induction transformation cultures with various concentrations. The proliferation of lymphocytes was measured by MTT method, effects of MSC and CBMNC on mixed lymphocyte response and PHA-induced lymphocyte transformation were investigated. The results showed that both 5 x 10(4), 1 x 10(4) MSC and 2 x 10(5) CBMNC could inhibit the mixed lymphocyte response (MLR) and PHA induced transformation. But with lower concentrations (MSC < or = 1 x 10(3), CBMNC < or = 1 x 10(5)), the inhibition effects of MSC and CBMNC were less consistent. 1 x 10(2) MSC and 1 x 10(4) CBMNC mainly increased the lymphocyte activation. In addition, the inhibition ratio of 5 x 10(4) MSC (MLC 65.3%, PHA 79.1%) was higher than that of 2 x 10(5) CBMNC (MLC 8.6%, PHA 37.3%). It is concluded that larger numbers of both MSC and CBMNC showed negative immunomodulatory functions and inhibited the mixed lymphocyte response and induction of transformation in vitro. Moreover, the inhibitory effect of MSC was much stronger than that of CBMNC.