C-terminal deletion-induced condensation sequesters AID from IgH targets in immunodeficiency.

In activated B cells, activation-induced cytidine deaminase (AID) generates programmed DNA lesions required for antibody class switch recombination (CSR), which may also threaten genome integrity. AID dynamically shuttles between cytoplasm and nucleus, and the majority stays in the cytoplasm due to active nuclear export mediated by its C-terminal peptide. In immunodeficient-patient cells expressing mutant AID lacking its C-terminus, a catalytically active AID-delC protein accumulates in the nucleus but nevertheless fails to support CSR. To resolve this apparent paradox, we dissected the function of AID-delC proteins in the CSR process and found that they cannot efficiently target antibody genes. We demonstrate that AID-delC proteins form condensates both in vivo and in vitro, dependent on its N-terminus and on a surface arginine-rich patch. Co-expression of AID-delC and wild-type AID leads to an unbalanced nuclear AID-delC/AID ratio, with AID-delC proteins able to trap wild-type AID in condensates, resulting in a dominant-negative phenotype that could contribute to immunodeficiency. The co-condensation model of mutant and wild-type proteins could be an alternative explanation for the dominant-negative effect in genetic disorders.

Phytoplankton photosynthetic rate measurement using tunable pulsed light induced fluorescence kinetics.

The photosynthetic process of phytoplankton is the basis of the material circulation and energy flow of the ecosystem. The rapid and accurate measurement of phytoplankton photosynthesis rate is of great significance to water ecological environment monitoring, marine resource assessment and global climate change prediction. On the basis of "Bio-Optical" model, a photosynthetic rate measurement method based on tunable pulsed light induced fluorescence kinetics was put forward in this paper. The chlorophyll fluorescence was used as the probe of photosynthesis process, and the phytoplankton photosynthetic rate was evaluated by the photosynthetic electron transport rate. Comparative experiment results showed that the photosynthetic electron transport rate measured by fluorescence kinetic method under different conditions of DCMU, culture light and nutrients (nitrogen) were consistent with the photosynthetic oxygen evolution rate measured by oxygen evolution method, and the correlation coefficient R2 were 0.934, 0.957 and 0.955 respectively.

Carry-over effect between diet and physical activity: the bottom-up and top-down hypotheses of hierarchical self-efficacy.

Healthy diet and physical activity have consistently been found to be positively correlated; however, most health behavior theories are focused on regulation of changes in single, rather than multiple, behaviors. Thus, this study explored the mechanism of the carry-over effect between diet and physical activity by conducting a longitudinal study with 706 participants to test the bottom-up and top-down hypotheses of hierarchical self-efficacy (SE). At Time 1 (baseline) and Time 3 (4 weeks after baseline), dietary behavior, physical activity, and self-efficacies of these behaviors (at the contextual level) were measured, while at Time 2 (2 weeks after baseline), general SE (at the general level) was assessed. Mediation analysis and structural equation models supported both the bottom-up and top-down hypotheses for different levels of self-efficacies, suggesting that hierarchical SE is an important factor underlying the carry-over mechanism between diet and physical activity.

[Study of the Impact of Sample Thickness on Thin Film Method X-Ray Fluorescence Spectrum Measurement].

The mixed samples of nylon film enrichment of Cr, Pb and Cd three elements and glass fiber membrane filter were as the research object. With the method of superposition of membrane filter, the XRF spectra were measured under different thin film samples thicknesses. According the changes of characteristic XRF of Cr, Pb and Cd elements in the mixed sample and Ca, As and Sr elements in glass fiber membranes, the effects of sample thickness on thin film method XRF spectrum measurement were studied. The study results showed that the effects of thin film sample thickness on the fluorescent properties of elements with characteristic spectral lines in different energy ranges were different. The energy of characteristic spectral lines was greater, the loss of element characteristic X-ray fluorescence when it passed through membrane and reached detector was less. But matrix effect caused by thin film sample thickness increase was stronger with the energy of characteristic spectral lines greater. The background fluorescent intensity in corresponding characteristic spectral line location was greater. So the impact of matrix effect caused by sample thickness increase on thin film method XRF spectrum measurement sensitivity was greater. For elements with low energy characteristic spectral lines (energy≤7 keV), the way of increasing thin film sample thickness in order to increase the mass-thickness concentration of component measured, can not effectively improve the sensitivity of thin film method XRF spectrum measurement. And thin film samples thickness within 0.96 mm is conductive to the measurement and analysis of XRF spectrum. For element with higher energy characteristic spectra lines(energy＞7 keV), the sensitivity of XRF spectrum measurement can be appropriately increased by the way of increase thin film sample thickness in order to increase the mass-thickness concentration of component measured. And thin film samples thickness within 0.96～2.24 mm is more conductive to the measurement and analysis of XRF spectrum. The study provides an important theoretical basis for thin sample preparation and enrichment technology of thin film method X-ray fluorescence spectrum analysis the atmosphere and water heavy metal.

[The study of selecting sample detecting position and lead plate inner material in thin film method X-ray fluorescence measurement].

(1) In this paper type 316 stainless steel metal plate as the research object, the selection of sample detecting position was studied when thin film method X-ray fluorescence measurement was conducted. The study showed that the optimal location for the sample detection was sample distance X-ray tube and detector baseline 1cm with the baseline into a 16°angle. (2) Heavy metal pollutants of Pb, Cd and Cr in industrial ambient air as the main analysis object, when thin film method X-ray fluorescence conducted with lead plate protection, X-rays will penetrate the membrane and continuely stimulate the protective lead plate. Therefore there is lead spectral line interference in the filter membrane background spectrum, which will affect the detection of lead element in real samples. Studies show that when a layer of isolating material was applied between the thin sample and the protective lead plate, the interference of lead line can effectively be avoided. (3) Several rigid insulating material of type 316 stainless steel, brass, aluminum, red copper and PTEE as lead inner material were selected and studied. The study results showed that compared with X-ray fluorescence spectra of other lead inner materials, the X-ray fluorescence spectrum of red copper contained the least element spectral lines. There were not Cr, Cd and Pb spectrum peaks in the X-ray fluorescence spectrum of red copper. And the target timber scattering spectrum intensity in the high energy part was weaker compared to other X-ray fluorescence spectrum. The above analysis shows that red copper has the minimal disturbance to the actual measurement of heavy metals Cr, Cd and Pb. At the same time, red copper as lead inner materials can effectively avoid the interference of lead spectrum line in lead plate. So red copper is the best lead plate inner materials in thin film method X-ray fluorescence spectroscopy measurement. This study provides an important theoretical basis for the assembling and setting'up air and water weight metal X-ray fluorescence spectrometer.

A skin-specific α-Synuclein seeding amplification assay for diagnosing Parkinson's disease.

The seeding amplification assay (SAA) has recently emerged as a valuable tool for detecting α-synuclein (αSyn) aggregates in various clinically accessible biospecimens. Despite its efficiency and specificity, optimal tissue-specific conditions for distinguishing Parkinson's disease (PD) from non-PD outside the brain remain underexplored. This study systematically evaluated 150 reaction conditions to identify the one with the highest discriminatory potential between PD and non-synucleinopathy controls using skin samples, resulting in a modified SAA. The streamlined SAA achieved an overall sensitivity of 92.46% and specificity of 93.33% on biopsy skin samples from 332 PD patients and 285 controls within 24 h. Inter-laboratory reproducibility demonstrated a Cohen's kappa value of 0.87 (95% CI 0.69-1.00), indicating nearly perfect agreement. Additionally, αSyn seeds in the skin were stable at -80 °C but were vulnerable to short-term exposure to non-ultra-low temperatures and grinding. This study thoroughly investigated procedures for sample preprocessing, seed amplification, and storage, introducing a well-structured experimental framework for PD diagnosis using skin samples.

Fork coupling directs DNA replication elongation and termination.

DNA replication is initiated at multiple loci to ensure timely duplication of eukaryotic genomes. Sister replication forks progress bidirectionally, and replication terminates when two convergent forks encounter one another. To investigate the coordination of replication forks, we developed a replication-associated in situ HiC method to capture chromatin interactions involving nascent DNA. We identify more than 2000 fountain-like structures of chromatin contacts in human and mouse genomes, indicative of coupling of DNA replication forks. Replication fork interaction not only occurs between sister forks but also involves forks from two distinct origins to predetermine replication termination. Termination-associated chromatin fountains are sensitive to replication stress and lead to coupled forks-associated genomic deletions in cancers. These findings reveal the spatial organization of DNA replication forks within the chromatin context.

A Sensitive Response Index Selection for Rapid Assessment of Heavy Metals Toxicity to the Photosynthesis of Chlorella pyrenoidosa Based on Rapid Chlorophyll Fluorescence Induction Kinetics.

Heavy metals as toxic pollutants have important impacts on the photosynthesis of microalgae, thus seriously threatening the normal material circulation and energy flow of the aquatic ecosystem. In order to rapidly and sensitively detect the toxicity of heavy metals to microalgal photosynthesis, in this study, the effects of four typical toxic heavy metals, chromium (Cr(VI)), cadmium (Cd), mercury (Hg), and copper (Cu), on nine photosynthetic fluorescence parameters (φPo, ΨEo, φEo, δRo, ΨRo, φRo, FV/FO, PIABS, and Sm) derived from the chlorophyll fluorescence rise kinetics (OJIP) curve of microalga Chlorella pyrenoidosa, were investigated based on the chlorophyll fluorescence induction kinetics technique. By analyzing the change trends of each parameter with the concentrations of the four heavy metals, we found that compared with other parameters, φPo (maximum photochemical quantum yield of photosystem II), FV/FO (photochemical parameter of photosystem II), PIABS (photosynthetic performance index), and Sm (normalized area of the OJIP curve) demonstrated the same monotonic change characteristics with an increase in concentration of each heavy metal, indicating that these four parameters could be used as response indexes to quantitatively detect the toxicity of heavy metals. By further comparing the response performances of φPo, FV/FO, PIABS, and Sm to Cr(VI), Cd, Hg, and Cu, the results indicated that whether it was analyzed from the lowest observed effect concentration (LOEC), the influence degree by equal concentration of heavy metal, the 10% effective concentration (EC10), or the median effective concentration (EC50), the response sensitivities of PIABS to each heavy metal were all significantly superior to those of φRo, FV/FO, and Sm. Thus, PIABS was the most suitable response index for sensitive detection of heavy metals toxicity. Using PIABS as a response index to compare the toxicity of Cr(VI), Cd, Hg, and Cu to C. pyrenoidosa photosynthesis within 4 h by EC50 values, the results indicated that Hg was the most toxic, while Cr(VI) toxicity was the lowest. This study provides a sensitive response index for rapidly detecting the toxicity of heavy metals to microalgae based on the chlorophyll fluorescence induction kinetics technique.

Sevoflurane participates in the protection of rat renal ischemia-reperfusion injury by down-regulating the expression of TRPM7.

INTRODUCTION: To investigate the protective effect of sevoflurane preconditioning on renal ischemia-reperfusion injury (renalischemiareperfusionmodel, RIRI) and its related mechanism. METHODS: Eighty healthy adult male SD rats were randomly divided into control group (Sham group), model group (RIRI group), sevoflurane pretreatment group (Sev group) and TRPM7 inhibitor combined with sevoflurane pretreatment group (T + Sev group), 20 animals in each group. Hematoxylin-eosin (HE) staining was used to observe the pathological changes of renal tissue, and the levels of creatinine and urea nitrogen in each group were detected. Deoxyribonucleic acid terminal transferase-mediated dUTP nick end labeling (TUNEL) assay was used to detect renal cell apoptosis, and Western blottingwas used to detect the expression of apoptotic proteins cleaved-caspase-3, bax, Bcl-2, and TRPM7 in renal tissue; Detection of oxidative stress-related index levels in renal tissue and levels of inflammatory factors in renal tissue and serum. RESULTS: Compared with the Sham group, the renal tissue pathological damage was aggravated, the levels of creatinine and blood urea nitrogen were increased, and the apoptosis was increased in the RIR group and the Sev group. Death, malondialdehyde (MDA) levels and inflammatory factors were increased, and superoxide dismutase (SOD) levels were decreased (all p < .05); The scores, apoptosis rate, MDA level, and relative expression of inflammatory factor levels were decreased, and SOD levels were increased (all p < .05). Compared with the Sev group, the renal tissue pathological damage in the T + Sev group was aggravated, creatinine, blood urea nitrogen levels increased, apoptosis increased, apoptosis-related proteins cleaved-caspase-3, bax, Bcl-2 showed increased apoptosis, malondialdehyde (MDA) levels, inflammatory factor levels increased, ultrahigh The levels of oxide dismutase (SOD) were decreased (all p < .05). CONCLUSIONS: Therefore, we believe that sevoflurane is involved in the protection of rat renal ischemia-reperfusion injury by downregulating the expression of TRPM7.

Cohesin maintains replication timing to suppress DNA damage on cancer genes.

Cohesin loss-of-function mutations are frequently observed in tumors, but the mechanism underlying its role in tumorigenesis is unclear. Here, we found that depletion of RAD21, a core subunit of cohesin, leads to massive genome-wide DNA breaks and 147 translocation hotspot genes, co-mutated with cohesin in multiple cancers. Increased DNA damages are independent of RAD21-loss-induced transcription alteration and loop anchor disruption. However, damage-induced chromosomal translocations coincide with the asymmetrically distributed Okazaki fragments of DNA replication, suggesting that RAD21 depletion causes replication stresses evidenced by the slower replication speed and increased stalled forks. Mechanistically, approximately 30% of the human genome exhibits an earlier replication timing after RAD21 depletion, caused by the early initiation of >900 extra dormant origins. Correspondingly, most translocation hotspot genes lie in timing-altered regions. Therefore, we conclude that cohesin dysfunction causes replication stresses induced by excessive DNA replication initiation, resulting in gross DNA damages that may promote tumorigenesis.

PEM-seq comprehensively quantifies DNA repair outcomes during gene-editing and DSB repair.

The repair products of double-stranded DNA breaks (DSBs) are crucial for investigating the mechanism underlying DNA damage repair as well as evaluating the safety and efficiency of gene-editing; however, a comprehensively quantitative assay remains to be established. Here, we describe the step-by-step instructions of the primer extension-mediated sequencing (PEM-seq), followed by the framework of data processing and statistical analysis. PEM-seq presents a full spectrum of repair outcomes for both genome-editing-induced and endogenous DSBs in mouse and human cells. For complete details on the use and execution of this profile, please refer to Gan et al. (2021), Yin et al. (2019), Liu et al. (2021a), and Zhang et al. (2021).

A new method for detecting mixed bacteria based on multi-wavelength transmission spectroscopy technology.

Previously, we successfully realized the identification of a single species of bacteria based on the multi-wavelength transmission spectrum of bacteria. The current research is focused on realizing the spectral analysis of mixed bacteria. Principal component analysis-Monte Carlo (PCA-MC) model was developed for the implementation of spectral separation of mixed bacteria by obtaining the ratio of components. And, the separated spectrum was regarded as the model input of the neural network concentration inversion model to obtain the concentration of each bacteria in the mix. Mean relative errors in component analysis of mixing S.aureus with K.pneumoniae, mixing S.aureus with S.typhimurium twice, mixing K.pneumoniae with S.typhimurium are 3%, 2%, 3.9% and 6.1%, respectively. The coefficient of determination (R2) of validation set and test set are 0.9947 and 0.9954 in concentration inversion model. The results show that this method can quickly and accurately determine the component ratio and concentration information in the mixed bacteria. A new method was proposed to separate the spectrum of mixed bacteria effectively and measure its concentration quickly, which makes a big step forward in the detection and online monitoring of waterborne microbial contamination based on multi-wavelength transmission spectroscopy.

RAG2 abolishes RAG1 aggregation to facilitate V(D)J recombination.

RAG1 and RAG2 form a tetramer nuclease to initiate V(D)J recombination in developing T and B lymphocytes. The RAG1 protein evolves from a transposon ancestor and possesses nuclease activity that requires interaction with RAG2. Here, we show that the human RAG1 aggregates in the nucleus in the absence of RAG2, exhibiting an extremely low V(D)J recombination activity. In contrast, RAG2 does not aggregate by itself, but it interacts with RAG1 to disrupt RAG1 aggregates and thereby activate robust V(D)J recombination. Moreover, RAG2 from mouse and zebrafish could not disrupt the aggregation of human RAG1 as efficiently as human RAG2 did, indicating a species-specific regulatory mechanism for RAG1 by RAG2. Therefore, we propose that RAG2 coevolves with RAG1 to release inert RAG1 from aggregates and thereby activate V(D)J recombination to generate diverse antigen receptors in lymphocytes.

Molecularly imprinted polymer coated Mn-doped ZnS quantum dots embedded in a metal-organic framework as a probe for selective room temperature phosphorescence detection of chlorpyrifos.

As one of the most widely used organophosphorus pesticides, chlorpyrifos (CPF) is toxic to humans. However, the rapid, effective and sensitive detection of CPF is still a challenge. In this paper, a novel molecularly imprinted phosphorescent sensor with a core-shell structure (Mn:ZnS QDs@ZIF-8@MIP) using Mn:ZnS quantum dots (QDs) as phosphorescent emitters was prepared for the highly sensitive and selective detection of CPF, and a simple and rapid room-temperature phosphorescence (RTP) detection method for CPF was proposed. For the prepared Mn:ZnS QDs@ZIF-8@MIP, Mn:ZnS QDs had good phosphorescence emission characteristics, ZIF-8 as support materials was used to improve the dispersibility of Mn:ZnS QDs, and molecularly imprinted polymer (MIP) on the surface of ZIF-8 was used to improve the selectivity of Mn:ZnS QDs for CPF. Under the optimal response conditions, the RTP intensity of Mn:ZnS QDs@ZIF-8@MIP showed a rapid response to CPF (less than 5 min), the RTP intensity ratio of P 0/P had a good linear relationship with the concentration of CPF in the range of 0-80 µM, and the detection limit of this method was 0.89 µM with the correlation coefficient of 0.99. Moreover, this simple and rapid method has been successfully used to detect CPF in real water samples with satisfactory results, and the recoveries ranged from 92% to 105% with a relative standard deviation of less than 1%. This method combines the advantages of phosphorescence emission and molecular imprinting, and greatly reduces the potential interferences of competitive substances, background fluorescence and scattered light, which opens up a broad prospect for the highly sensitive and selective detection of pollutants in water based on molecularly imprinted phosphorescent sensors.

Artificial neural networks combined multi-wavelength transmission spectrum feature extraction for sensitive identification of waterborne bacteria.

Present research is focused on the rapid and accurate identification of bacterial species based on artificial neural networks combined with spectral data processing technology. The spectra of different bacterial species in the logarithmic growth phase were obtained. Model input features were extracted from the raw spectra using signal processing techniques, including normalization, principal component analysis (PCA) and area-based feature value extraction. The identification models based on artificial neural network of back propagation neural networks (BPNN), generalized regression neural networks (GRNN) and probabilistic neural networks (PNN) were developed using the extracted features in order to ascertain whether the different species of bacteria could be differentiated. The performance of developed models and its corresponding signal processing techniques is tested by the recognition accuracy of validation set and test set, and model error. The maximum recognition accuracy of normalized spectrum combined with BPNN was 95.5% (error: 10%, test accuracy: 100%). The total recognition accuracy of PCA-reduced features (200-400 nm) combined with GRNN resulted in 96.3%~96.8% (error: 3.3%~6.7%, test accuracy: 97.5%~100%). While the overall recognition accuracy of area-based features combined with GRNN reached 97.3% with test accuracy of 100% (model error: 5.0%). Choosing of model and signal processing techniques has a positive influence on improving classification accuracy, so as to make it possible to realize the rapid detection and online monitoring of waterborne microbial contamination.

Transcription shapes DNA replication initiation to preserve genome integrity.

BACKGROUND: Early DNA replication occurs within actively transcribed chromatin compartments in mammalian cells, raising the immediate question of how early DNA replication coordinates with transcription to avoid collisions and DNA damage. RESULTS: We develop a high-throughput nucleoside analog incorporation sequencing assay and identify thousands of early replication initiation zones in both mouse and human cells. The identified early replication initiation zones fall in open chromatin compartments and are mutually exclusive with transcription elongation. Of note, early replication initiation zones are mainly located in non-transcribed regions adjacent to transcribed regions. Mechanistically, we find that RNA polymerase II actively redistributes the chromatin-bound mini-chromosome maintenance complex (MCM), but not the origin recognition complex (ORC), to actively restrict early DNA replication initiation outside of transcribed regions. In support of this finding, we detect apparent MCM accumulation and DNA replication initiation in transcribed regions due to anchoring of nuclease-dead Cas9 at transcribed genes, which stalls RNA polymerase II. Finally, we find that the orchestration of early DNA replication initiation by transcription efficiently prevents gross DNA damage. CONCLUSION: RNA polymerase II redistributes MCM complexes, but not the ORC, to prevent early DNA replication from initiating within transcribed regions. This RNA polymerase II-driven MCM redistribution spatially separates transcription and early DNA replication events and avoids the transcription-replication initiation collision, thereby providing a critical regulatory mechanism to preserve genome stability.

Simultaneous determination of nitrate, chemical oxygen demand and turbidity in water based on UV-Vis absorption spectrometry combined with interval analysis.

In this paper, a new method for simultaneous determination of nitrate, COD and turbidity in water based on UV-Vis absorption spectrometry combined with interval analysis was studied. By analyzing the spectral absorption characteristics of nitrate, COD, and turbidity standard solutions and the mixtures of them, the absorption spectra in the range of 225-260 nm, 260-320 nm and 320-700 nm were selected as the characteristic spectra of nitrate, COD and turbidity, respectively. Multiplicative scatter correction was employed to compensate turbidity of the absorption spectra of the mixture solutions in the wavelength range of 225-320 nm. Then, the spectra after turbidity compensation in the range of 225-260 nm was compensated for COD using the method of spectral difference. The original spectra in the range of 320-700 nm, the turbidity compensated spectra in the range of 260-320 nm, and the COD compensated spectra in the range of 225-260 nm were analyzed by PLS algorithm in order to calculate the concentrations of nitrate, COD and turbidity in the mixture solutions. The results showed that this method could simultaneously and accurately determine the concentrations of nitrate, COD and turbidity. After interval analysis, all the correlation coefficients (R2) between the predicted values and the true values of nitrate, COD and turbidity were higher than 0.9, and root mean square error (RMSE) of predicted values were between 0.696 and 2.337.

Rapid and Sensitive Detection of Water Toxicity Based on Photosynthetic Inhibition Effect.

To achieve rapid and sensitive detection of the toxicity of pollutants in the aquatic environment, a photosynthetic inhibition method with microalgae as the test organism and photosynthetic fluorescence parameters as the test endpoint was proposed. In this study, eight environmental pollutants were selected to act on the tested organism, Chlorella pyrenoidosa, including herbicides (diuron, atrazine), fungicides (fuberidazole), organic chemical raw materials (phenanthrene, phenol, p-benzoquinone), disinfectants (trichloroacetonitrile uric acid), and disinfection by-products (trichloroacetonitrile). The results showed that, in addition to specific PSII inhibitors (diuretic and atrazine), other types of pollutants could also quickly affect the photosynthetic system. The photosynthetic fluorescence parameters (Fv/Fm, Yield, α, and rP) could be used to detect the effects of pollutants on the photosynthetic system. Although the decay rate of the photosynthetic fluorescence parameters corresponding to the different pollutants was different, 1 h could be used as an appropriate toxicity exposure time. Moreover, the lowest respondent concentrations of photosynthetic fluorescence parameters to diuron, atrazine, fuberidazole, phenanthrene, P-benzoquinone, phenol, trichloroacetonitrile uric acid, and trichloroacetonitrile were 2 µg·L-1, 5 µg·L-1, 0.05 mg·L-1, 2 µg·L-1, 1.0 mg·L-1, 0.4 g·L-1, 0.1 mg·L-1, and 2.0 mg·L-1, respectively. Finally, diuron, atrazine, fuberidazole, and phenanthrene were selected for a comparison of their photosynthetic inhibition and growth inhibition. The results suggested that photosynthetic inhibition could overcome the time dependence of growth inhibition and shorten the toxic exposure time from more than 24 h to less than 1 h, or even a few minutes, while, the sensitivity of the toxicity test was not weakened. This study indicates that the photosynthetic inhibition method could be used for rapid detection of the toxicity of water pollutants and that algae fluorescence provides convenient access to toxicity data.

Transdermal iontophoresis delivery system for terazosin hydrochloride: an in vitro and in vivo study.

This study aimed to construct a transdermal iontophoresis delivery system for terazosin hydrochloride (IDDS-TEH), which included a positive and negative electrode hydrogel prescription. Intact guinea pig skin was used as a model for the skin barrier function, and the current intensity, terazosin hydrochloride (TEH) concentration, pH, competitive salt, and transdermal enhancer properties were studied. The blood drug concentration was determined in Sprague-Dawley (SD) rats using HPLC, and the antihypertensive effects of IDDS-TEH were evaluated in spontaneously hypertensive rats (SHRs). The results showed that the steady-state penetration rate of TEH increased (from 80.36 µg·cm-2·h-1 to 304.93 µg·cm-2·h-1), followed by an increase in the current intensity (from 0.10 mA·cm-2 to 0.49 mA·cm-2). The pH values also had a significant influence on percutaneous penetration. The blood concentration of IDDS-TEH was significantly higher (p < .05) than with passive diffusion, which could not be detected. The main pharmacokinetic parameters of the high current group (0.17 mA·cm-2) and the low current group (0.09 mA·cm-2) were AUC0-t: 5873.0 ng·mL-1·h and 2493.7 ng·mL-1·h, respectively. Meanwhile, the pharmacodynamic results showed that IDDS-TEH significantly decreased the blood pressure of SHRs compared with the TEH hydrogel without loading current. Therefore, TEH could be successfully delivered by the transdermal iontophoresis system in vitro and in vivo, and further clinical studies should be explored to develop a therapeutically useful protocol.

A new open tubular capillary microextraction and sweeping for the analysis of super low concentration of hydrophobic compounds.

A sample pre-concentration method based on the in-line coupling of in-tube solid-phase microextraction and electrophoretic sweeping was developed for the analysis of hydrophobic compounds. The sample pre-concentration and electrophoretic separation processes were simply and sequentially carried out with a (35%-phenyl)-methylpolysiloxane-coated capillary. The developed method was validated and applied to enrich and separate several pharmaceuticals including loratadine, indomethacin, ibuprofen and doxazosin. Several parameters of microextration were investigated such as temperature, pH and eluant. And the concentration of microemulsion that influences separation efficiency and microextraction efficiency were also studied. Central composite design was applied for the optimization of sampling flow rate and sampling time that interact in a very complex way with each other. The precision, sensitivity and recovery of the method were investigated. Under the optimal conditions, the maximum enrichment factors for loratadine, indomethacin, ibuprofen and doxazosin in aqueous solutions are 1355, 571, 523 and 318, respectively. In addition, the developed method was applied to determine loratadine in rabbit blood sample.