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Background: Diabetic encephalopathy (DE) is considered as one of the complications of diabetes,which is associated with cognitive impairment in the pathological process of development. Up to now, phospholipid phosphatase related 4 (Plppr4), also known as plasticity related gene 1 (PRG-1) has been revealed its important role in neuroplasticity. However, the underlying mechanisms of Plppr4 on the basis of diabetic-induced cognitive dysfunction (DCD) are still unknown. The aim of current study was to provide insight into molecular mechanism and cellular heterogeneity underlying DCD, and investigate the functional role of PRG-1 involved in this process., Methods: Combined Single-cell RNA sequencing (scRNA-seq) and RNA transcriptome analysis, the distinct sub-populations, functional heterogeneity as well as potential enriched signaling pathways of hippocampal cells could be elucidated., Results: We identified the sub-cluster of type I spiral ganglion neurons expressed marker gene as Amigo2 in cluster8 and Cnr1 in cluster 9 of hippocampal cells from DCD and the effect of those on neuronal cells interaction. We also found that PRG-1 was involved in the synaptic plasticity regulation of hippocampus via NGF\BDNF-Trkb signaling pathway. In high glucose induced HT22 cells injury model in vitro, we investigated that down-regulated PRG-1 along with down-regulated BDNF and also decreased expression of synapsin-1, PSD-95, SYN which are related to synaptic plasticity; Meanwhile, the Prg-1 targeted miR-18a-LncRNA NONRATG-022419 pairs related with significantly down-regulated expression of PRG-1., Conclusion: This study revealed the synaptic plasticity regulation of PRG-1 in DCD, and might provide the therapeutic target and potential biomarkers for early interventions in DCD patients., Competing Interests: Declarations. Ethics approval and consent to participate: N/A. Consent for publication: Not applicable. Competing interests: The authors declare no competing interests., (© 2025. The Author(s).)

Chronic hyperglycemia can cause changes in synaptic plasticity of hippocampal cells, which has accelerated the pathological process of cognitive dysfunction. However, the heterogeneity of the hippocampal cell populations under long term high glucose statement remains largely unknown. To mimic chronic hyperglycemia induced cognitive function deficit in vivo, db -/- diabetic mice was selected and Novel Object Recognition(NOR) behavior tests were performed. Based on diabetic induced cognitive impairment(CI) animal model, single-cell RNA sequencing was performed in the hippocampus of CI group (21,379 cells) or control group (20,045 cells), and single cell RNA sequencing was applied, and then the single cell atlas of gene expression was profiled. The comprehensive analysis explicated 18 nerve cell clusters, including 9 distinct sub-clusters, More in-depth analysis of oligodendrocyte precursor cells(OPCs) showed five distinct OPCs sub-clusters including expressing marker gene Lingo2-OPCs, Kcnc1-OPCs, Sst-OPCs, Slc6a1-OPCs and Lhfpl3-OPCs, which seems to be able to proliferate, migrate, and finally differentiate into mature oligodendrocytes and produce myelin. To be noted, differentially expressed genes(DEGs) of the Sst-OPCs sub-cluster indicated that the genes participating in neuroactive ligand-receptor interaction, nervous system development and inflammatory process were up-regulated in diabetic induced cognitive impairment(DCI) groups compared to normal control groups. Integrating the data of neuroplasticity regulation, the 20th top-enriched biological process was associated with neuroplasticity regulation in CI groups compared to control groups. Among these neuroplasticity-related genes, the intersectional gene Sstr2 may play an important role in neuroplasticity regulation. Focused on neuroplasticity regulation and its related specific genes may provide potential new clues for the treatment of diabetes mellitus complicated with cognitive impairment. In summary, we showed the comprehensively transcriptional landscape of hippocampal cells in the db -/- diabetic mice with cognitive dysfunction, distinctive cell sub-clusters and the gene expression characteristics were identified, and also their special functions were proposed, which may give new clues and potential targets for diagnosis and treatment of diabetic encephalopathy., Competing Interests: Declaration of Competing Interest All of the author has no conflicts of interest to declare., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Objectives: Chronic hyperglycemia is considered as an important factor to promote the neurodegenerative process of brain, and the synaptic plasticity as well as heterogeneity of hippocampal cells are thought to be associated with cognitive dysfunction in the early process of neurodegeneration. To date, fibronectin type III domain-containing protein 5 (FNDC5) has been highlighted its protective role in multiple neurodegenerative diseases. However, the potential molecular and cellular mechanisms of FNDC5 on synaptic plasticity regulation in cognitive impairment (CI) induced by diabetics are still need to known., Methods/design: To investigate the heterogeneity and synaptic plasticity of hippocampus in animals with CI state induced by hyperglycemia, and explore the potential role of FNDC5 involved in this process. Firstly, the single cell sequencing was performed based on the hippocampal tissue from db diabetic mice induced CI and normal health control mice by ex vivo experiments; and then the integrated analysis and observations validation using Quantitative Real-time PCR, western blot as well as other in vitro studies., Results: We observed and clarified the sub-cluster of type IC spiral ganglion neurons expressed marker genes as Trmp3 and sub-cluster of astrocytes with marker gene as Atp1a2 in hippocampal cells from diabetic animals induced CI and the effect of those on neuron-glial communication. We also found that FNDC5\BDNF-Trk axis was involved in the synaptic plasticity regulation of hippocampus. In high glucose induced brain injury model in vitro, we investigated that FNDC5 significantly regulates BDNF expression and that over-expression of FNDC5 up-regulated BDNF expression (p < 0.05) and can also significantly increase the expression of synapsin-1 (p < 0.05), which is related to synaptic plasticity, In addition, the unbalanced methylation level between H3K4 and H3K9 in Fndc5 gene promoter correlated with significantly down-regulated expression of FNDC5 (p < 0.05) in the hyperglycemia state., Conclusion: The current study revealed that the synaptic plasticity of hippocampal cells in hyperglycemia might be regulated by FNDC5\BDNF-Trk axis, playing the protective role in the process of CI induced by hyperglycemia and providing a target for the early treatment of hyperglycemia induced cognitive dysfunction in clinic., (© 2023 John Wiley & Sons Ltd.)

Gap junction (GJ) is a special cell membrane structure composed of connexin. Connexin is widely distributed and expressed in all tissues except differentiated skeletal muscle, red blood cells, and mature sperm cells, which is related to the occurrence of many genetic diseases due to its mutation. Its function of regulating immune response, cell proliferation, migration, apoptosis, and carcinogenesis makes it a therapeutic target for a variety of diseases. In this paper, the possible mechanism of its action in nervous system-related diseases and treatment are reviewed., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

Von Willebrand factor (VWF) is a glycoprotein synthesized and secreted by vascular endothelial cells and megakaryocytes, found on plasma surface, endothelial cells, and α-granule of platelets. VWF can be interacted with collagen and platelet membrane glycoproteins GPIb and GPIb-IIa and play an important role in platelet adhesion and aggregation. Growing research evidence suggests that VWF also mediates the prevention or protesting of hepatocellular carcinoma (HCC) in chronic hepatitis B (CHB) patients from several clinical studies. While the mechanism of VWF in HCC protection or protest is still unclear, further study is required. This article aims to rationalize the role of VWF in the development of HCC, and the functional domain of VWF in cancer as well as cross-talking with platelets and miRNAs. This article also looks forward to the future development and challenges of VWF research., Competing Interests: The authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article., (© The Author(s), 2022.)

Introduction: Our previous work has shown that somatostatin effectively inhibits neuropathic pain by activating its type 2 receptor (SSTR2) in the dorsal root ganglion (DRG) and spinal cord of mice. However, the underlying mechanism of this activation has not been elucidated., Methods: To explore further mechanisms, we examined pain behavior and the expression of neuropeptides such as calcitonin gene-related peptide (CGRP) in dorsal root ganglion neurons(DRGs) as well as the changes of the number of CGRP-IR DRGs in the mouse model of sciatic pinch nerve injury., Results: In this model, the number of medium and small DRG neurons in ipsilateral CGRP-IR was slightly increased, but not significantly, compared with sham animals at 3, 7, and 9 days after pinch nerve injury. This correlated with the behavioral readouts of hypersensitivity at the same time points. However, the magnitude of the painful behavior (Autotomy) was observed after application of SSTR2 antagonist (CYN154806, 5 mg/kg) in the injured nerve groups compared to the saline-treated injured group as well as the sham-operated group. Following pinch nerve injury, there was a significant decrease in the number of ipsilateral CGRP-IR small and medium DRG neurons in SSTR2 antagonist (anti-SSTR2)- but not saline-treated mice. These data also correlated with painful behavioral readouts where hypersensitivity was significantly increased by anti-SSTR2 but not saline treatment., Discussion/conclusion: In all, application of the SSTR2 antagonist to the pinched sciatic nerve suppressed CGRP expression and aggravated painful behavior, suggesting that CGRP expression in DRG neurons can be an important component of the pain mechanism and an indicator of pain behavior., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Phenolic compounds, the most abundant secondary metabolites in plants, have received more and more attention in recent years because of their distinct bioactivities. This review summarizes different types of phenolic compounds and their extraction and analytical methods used in the recent reports, involving 59 phenolic compounds from 52 kinds of plants. The extraction methods include solid-liquid extraction, ultrasound-assisted extractions, microwave-assisted extractions, supercritical fluid extraction, and other methods. The analysis methods include spectrophotometry, gas chromatography, liquid chromatography, thin-layer chromatography, capillary electrophoresis, and near-infrared spectroscopy. After illustrating the specific conditions of the analytical methods, the advantages and disadvantages of each method are also summarized, pointing out their respective suitability. This review provides valuable reference for identification and/or quantification of phenolic compounds from natural products., (Copyright © 2017 China Pharmaceutical University. Published by Elsevier B.V. All rights reserved.)

Limnological studies have long focused on the nine largest plateau lakes and ecological assessment of lake eutrophication in Yunnan. However, comprehensive surveys of small and me-dium-sized lakes and their response to multiple stressors are largely absent. Here, we combined multi-proxy sediment analyses and documentary data, aiming to identify the long-term pattern of diatom community changes under the influence of climate change and anthropogenic disturbances, and to quantify their driving strengths. The results showed that Xihu Lake was in a pre-impact state with little ecological variability before the 1950s. Thereafter, expansion of land use increased the nutrient input and altered the lake hydrology, resulting in a significant shift of dominant diatoms from Cocconeis placentula to Fragilaria spp. From 1997, accelerating nutrient enrichment and hydrological changes led to the replacement of benthic diatoms by planktonic ones, resulting in a consistent loss of macrophytes and a decrease of ecological stability. Therefore, the types and strength of catchment development should be fully evaluated for effective protection of small and medium-sized alpine lakes in the context of global warming.

Objective To evaluate the feasibility and effectiveness of secundum atrial septal defect(ASD)occlusion with the septal occluder through right-chest small incision. Methods The clinical data of 140 secundum ASD patients (47 males and 93 females) aged 3-63 years who were treated in our center from August 2004 to July 2014 were retrospectively analyzed. The diameter of ASD was 6 to 36 mm. Under general anesthesia, all patients underwent intraoperative transtsophageal echocardiography (TEE), during which no associated cardiac deformity was found. All patients received ASD occlusion via a small incision (3-4 cm) at the right anterior chest. The occluders were released with the help of TEE. Results The atrial septal defect closure was successfully completed in 134 cases. Six cases received surgical closure of ASD after the failure of occlusion. The reasons of conversion included postoperative dislodgement of occlusion device (n=2, both were central type with large size) and technically unsuitable for occlusion (n=4, in whom residual shunt was found in 2 case, sieve pore type in 1 case, and intraoperative dislodgement in 1 case). All of these 6 patients were treated surgically under cardiopulmonary bypass. No dislocation of the device or atrial shunt was found within 3 to 48 months after the operation. Conclusion Occlusion via small chest incision of ASD under TEE guidance without cardiopulmonary bypass is a safe, minimally invasive, effective, and convenient treatment and worth clinical application.

Aim: Saquinavir (SQV) is the first protease inhibitor for the treatment of HIV infection, but with poor solubility. The aim of this study was to prepare a colloidal nanocrystal suspension for improving the oral absorption of SQV., Methods: SQV nanocrystals were prepared using anti-solvent precipitation-high pressure homogenization method. The nanocrystals were characterized by a Zetasizer and transmission electron microscopy (TEM). Their dissolution, cellular uptake and transport across the human colorectal adenocarcinoma cell line (Caco-2) monolayer were investigated. Bioimaging of ex vivo intestinal sections of rats was conducted with confocal laser scanning microscopy. Pharmacokinetic analysis was performed in rats administered nanocrystal SQV suspension (50 mg/kg, ig), and the plasma SQV concentrations were measured with HPLC., Results: The SQV nanocrystals were approximately 200 nm in diameter, with a uniform size distribution. The nanocrystals had a rod-like shape under TEM. The dissolution, cellular uptake, and transport across a Caco-2 monolayer of the nanocrystal formulation were significantly improved compared to those of the coarse crystals. The ex vivo intestinal section study revealed that the fluorescently labeled nanocrystals were located in the lamina propria and the epithelium of the duodenum and jejunum. Pharmacokinetic study showed that the maximal plasma concentration (Cmax) was 2.16-fold of that for coarse crystalline SQV suspension, whereas the area under the curve (AUC) of nanocrystal SQV suspension was 1.95-fold of that for coarse crystalline SQV suspension., Conclusion: The nanocrystal drug delivery system significantly improves the oral absorption of saquinavir.

Subtropical plateau lakes in Southeast Yunnan are known to have experienced multiple environmental stressors (lake pollution, eutrophication, land reclamation and droughts) during the past decades. Here, we applied multi-proxy analyses (i.e., diatom, grain size, C and N isotopic and chronology) of lake sediments, aiming to track the history of lake environment change and the characteristics of diatom community responses in Datun Lake for the past century. The results proved that the sedimentary diatom community records showed significant changes with the replacement of Fragilaria construens by Achnanthes minutissima. Combining with multiple-proxy records (i.e., isotopic data) and modern monitoring records, the ordination analyses and variation partitioning further suggested that industrial pollution and eutrophication were the major driving factors that led to the long-term shift of diatom community. In addition, the grain size results and meteorological data revealed that the reductions in hydrodynamic and water exchange intensity associated with damming and the occurrence of extreme droughts, thus led to the corresponding diatom community change.

Objective: To study the preparation of 20(S)-protopanaxadiol-phospholipid (20(S)-PPD) complex HAP assemblies., Methods: 20(S)-PPD phospholipid complex was assembled with the drug carriers of HAP. Effects of technological factors on the assembled amount were investigated, including HAP species, phospholipid complex concentration, ratio of HAP and assembled liquid, and then the preparation technology of 20(S)-PPD phospholipid complex HAP assemblies was determined., Results: The average quality of phospholipid complex assembled was 136. 26 mg/g,the average assembled rate was 5.3% for 20(S)-PPD phospholipid complex HAP assemblies prepared by the determined assembly process. FT-IR showed that 20(S)-PPD phospholipid complex was absorbed in the HAP assemblies, and the hydrogen-bonding effect was the main mechanism of HAP assemblies to assemble and adsorb phospholipid complex. The cumulative release rate in pH 7.4 phosphate buffer of the HAP assemblies indicated that the assemblies had the effect of sustained release., Conclusion: The phospholipid complex HAP assembles have the advantages of simple preparation process, and sustaining release effect, which can provide preliminary research foundation for research and development of 20(S)-PPD sustained-release preparations.

Yeputaoteng is the dried ground part of Ampelopsis sinica (Miq.) W.T. Wang, which is widely used in traditional Chinese medicine for preventing and treating tumors, chronic nephritis, hepatitis, rubella, traumatic bleeding, stomach heat and vomiting. A simple and reliable method using high-performance liquid chromatography with diode array detection (HPLC-DAD) was developed for the simultaneous determination of dihydromyricetin and resveratrol in Yeputaoteng. The chromatographic analysis was performed on a Dikma C18 column (250 × 4.6 mm, 5 µm) at 30°C with a gradient elution of acetonitrile and 0.1% phosphoric acid at a flow rate of 1 mL/min, and used ultraviolet detection at 292 and 306 nm. The established method was validated in terms of linearity, precision, reproducibility, stability and recovery. The calibration curves showed good linear regression (R(2) > 0.9994). Limits of detection and quantification fell in the ranges of 1.47-2.48 and 2.93-4.97 µg/mL, respectively. The mean recovery of dihydromyricetin and resveratrol was 104.1% [relative standard deviation (RSD): 2.94%] and 100.8% (RSD: 2.80%), respectively. The quantitative results indicated that the HPLC-DAD method can be effectively applied to the quality control of Yeputaoteng and its preparations.

A sensitive and specific method was developed and validated for the quantitation of one major metabolite of genipin in rats plasma. The major metabolite was isolated from rat bile via semi-preparative HPLC technology and its chemical structure was identified as genipin-1-o-glucuronic acid (GNP-GLU), which was for the first time used as a standard compound for quantitative analysis in rat plasma after administration of genipin. The application of high-performance liquid chromatography-tandem mass spectrometry in negative mode in multiple reaction monitoring mode was investigated. Chromatographic separation was achieved on an Eclipse XDB-C18 column using a mobile phase consisting of water with 0.1% formic acid (A)-acetonitrile (B). The limit of detecation was 0.214 ng/mL and the lower limit of quantification was 0.706 ng/mL. The calibration curve was linear from 1.27 to 3810 ng/mL for plasma samples, with a correlation coefficient of 0.9924. The intra- and inter-day precisions and accuracy were all within 15%. The recoveries of GNP-GLU and puerarin were above 90.0 and 76.2%, respectively. The highly sensitive method was successfully applied to estimate pharmacokinetic parameters of GNP-GLU following oral and intravenous administration of genipin to rats., (Copyright © 2013 John Wiley & Sons, Ltd.)

Glucuronidation and sulfation represent two major pathways in phase II drug metabolism in humans and other mammalian species. The great majority of drugs, for example, polyphenols, flavonoids and anthraquinones, could be transformed into sulfated and glucuronidated conjugates simultaneously and extensively in vivo. The pharmacological activities of drug conjugations are normally decreased compared with those of their free forms. However, some drug conjugates may either bear biological activities themselves or serve as excellent sources of biologically active compounds. As the bioactivities of drugs are thought to be relevant to the kinetics of their conjugates, it is essential to study the pharmacokinetic behaviors of the conjugates in more detail. Unfortunately, the free forms of drugs cannot be detected directly in most cases if their glucuronides and sulfates are the predominant forms in biological samples. Nevertheless, an initial enzymatic hydrolysis step using β-glucuronidase and/or sulfatase is usually performed to convert the glucuronidated and/or sulfated conjugates to their free forms prior to the extraction, purification and other subsequent analysis steps in the literature. This review provides fundamental information on drug metabolism pathways, the bio-analytical strategies for the quantification of various drug conjugates, and the applications of the analytical methods to pharmacokinetic studies., (Copyright © 2013 John Wiley & Sons, Ltd.)

Objective: To optimize the extraction process and macroporous resin for purification of Timosaponin B II from Anemarrhena asphodeloides., Methods: Orthogonal design L9 (34) was employed to optimize the circumfluence extraction conditions by taking the extraction yield of Timosaponin B II as index. The absorption-desorption characteristics of eight kinds of macroporous resins were evaluated, then the best resin was chosen to optimize the purification process conditions., Results: The optimum extraction conditions were as follows: the herb was extracted for 2 times (2 hours each time) with 8.5-fold 50% ethanol at the first time and 6-fold 50% ethanol at the second time. HPD100 resin showed a good property for the absorption-desorption of Timosaponin B II. The optimum technological conditions of HPD100 resin were as follows:the solution concentration was 0.23 mg/mL, the amount of saturated adsorption at 4/5 body volumn (BV) resin, the HPD100 resin was washed with 3 BV water and 6 BV 20% ethanol solution to remove the impurity, then the Timosaponin B II was desorbed by 5 BV ethanol solution. The purity of Timosaponin B II was about 50%., Conclusion: The optimized extraction process and purification is stable, efficient and suitable for industrial production.

The safety of geniposide, mainly focusing on its hepatotoxicity in rats, was determined by liver enzymes in serum and histopathology ultrastructural preparation. The lethal dose, 50% (LD50) of per oral geniposide was 1431.1 mg kg(-1). The acute toxicity study indicated geniposide at dose of 574 mg kg(-1) or more could cause hepatic toxicity in rats and the hepatotoxicity often appeared at 24-48 h after the oral administration. The hepatotoxicity was associated with oxidative stress with decrease of total superoxide dismutase activity and increase of malondialdehyde concentration in rats' livers. Subchronic toxicity study showed geniposide did not cause hepatotoxicity at the doses of 24.3 and 72.9 mg kg(-1) orally for 90 days in rats. Thus, acute hepatotoxicity of geniposide at high doses was likely to be linked to oxidative stress, while geniposide at normal dose of 24.3 mg kg(-1) or less did not cause hepatotoxicity even in the repeated dosing study.

A convenient and efficient one-pot synthesis of cyclopamine from peimisine is described. The key steps involve one-pot hydrazination and subsequent Bamford-Stevens reaction. The mild reaction conditions, high overall yield as well as an easy purification indicate this process can potentially be used for the scale-up preparation of cyclopamine., (Copyright © 2012 Elsevier B.V. All rights reserved.)

A sensitive high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for the quantification of genipin in rat plasma after hydrolysis with sulfatase. Genipin could not be detected directly as it could be transformed into other forms such as conjugated-genipin immediately after administration. The conjugated genipin could be hydrolyzed by sulfatase to genipin. The conditions of hydrolysis were investigated. Genipin and the internal standard, peoniflorin (IS), were separated on a reversed-phase column by gradient elution and detected using an electrospray ion source on a 4000 QTrap triple-quadrupole mass spectrometer. The quantification was performed using multiple reaction monitoring with selected precursor-product ion pairs of the transitions m/z 225.0 → 122.7 and m/z 479.1 → 449.1 for genipin and peoniflorin. The assay was linear over the concentration range of 1.368-1368 ng/mL, with correlation coefficients of 0.9989. Intra- and inter-day precisions and accuracy were all within 15%. The lower limit of quantification was 1.368 ng/mL. The recoveries of genipin and peoniflorin were more than 53.3 and 51.2%. The highly sensitive method was successfully applied to estimated pharmacokinetic parameters of genipin following oral and intravenous administration to rats. The absolute bioavailability of genipin was 80.2% in rat, which is the first report., (Copyright © 2011 John Wiley & Sons, Ltd.)

Objective: To research on the preparation of Arctigenin in vitro., Methods: Took enzyme concentration, time course and substrate concentration as investigation factors, used Box-Behnken design-response surface methodology to optimize the enzyme hydrolysis path of Arctigenin., Results: The best operational path for Arctigenin was as follows: the temperature was 50 degrees C, pH was 4.8, enzyme concentration was 0.44 U/mL, time course was 46.81 min, substrate concentration was 0.29 mg/mL, the conversion rate was 90.94%., Conclusion: This research can be regarded as a referencein preparing Arctigenin in vitro.

Objective: To optimize the process for purification of geniposide in the extract fluid of Gardenia jasminoides with macroporous absorption resin., Methods: By comparing the content and transfer ratio of geniposide during the process of purification, we optimized the process for purification of geniposide., Results: The optimal process for purification of geniposide with D301R macroporous absorption resin included the diameter height ratio 1:7.5, the concentration of the extract fluid of Gardenia jasminoides 2:1, the flow rate 1BV/h (1BV represented one column volume), the sample volume 1/3BV. We loaded the sample and left it for 2 hours, afterwards, rinsed the macroporous absorption resin using 2BV water until the solution became colourless. Then we rinsed the macroporous absorption resin with 20% alcohol,and the volume of elution solvent was 2BV. We collected 20% alcohol elution solvent and recycle alcohol using rotating evaporation and dried the rest solution in a vacuum to get the light yellow powder which was the purified geniposide., Conclusion: This process is simple and affordable, can be used to refine and purify geniposide in the extract fluid of Gardenia jasminoides Ellis. It provides a guidance for industrial production basis.

Objective: In order to obtain the optimal conditions for separating the alkaloids from the extract of Stemona japonica by selecting appropriate cation exchange resins., Methods: Seven types of cation exchange resins were evaluated in separating efficiency with measuring the adsorption ratio and eluting ratio of total alkaloids as indices, and the content of total alkaloids from Stemona japonica was determined as an index by spectrophotometry to choose the optimal technological parameters., Results: The optimal result of extraction was obtained as Stemona japonica shattered into thick powder, adding eight times amount of 90% alcohol and refluxing and extracting for 3 h (totally extracting for 3 times). Each gram of D004 cation exchange resin could absorb 0.5003 mg of the total alkaloid, and the desorption ratio was 68.45%. The transfer rate of total alkaloids was 58.70%. the product purity of alkaloids was up to 70%., Conclusion: The D004 cation exchange resin can be used for purificating total alkaloids from Stemona japonica and the established procedure is simple and feasible.

The cryptic plasmid pGY1, which is harbored by a clinical isolate of Salmonella enterica serovar Paratyphi A, was identified in a 9-year-old girl with paratyphoid fever in 2005, and its DNA sequence was determined. It is 3592 bp in length and had a G+C content of 43.3%. Three ORFs were predicted that share low similarity with hypothetical proteins in the GenBank database. pGY1 shared 36.6% sequence homology with the cryptic plasmid pIMVS1 from Salmonella typhimurium. Its unique sequence makes it attractive for further study to obtain insight into the evolutionary relationship of this plasmid with other Enterobacteriaceae plasmids.

Objective: To explore pharmacokinetic features of puerarin in pueraria spray and calculate pharmacokinetic parameters according to puerarin of drug-time curve in rabbits., Methods: The concentration of puerarin in plamsa was determined by HPLC. The methanol was used to sediment protine. The 3P87 program was used to calculate the pharmacokinetic parameters., Results: The vivo course of pueraria in spray could be described by the two compartment model. The main pharmacokinetic parameters of Pueraria spray were: t(l/2(beta)) =0.93 h, CL =44.23 mg x L(-1), AUC = 16.28 mg x h x L(-1), Cmax =5.9 mg x L(-1) and tmax = 0.975 h., Conclusion: The study will provide some scientific basises for the quality evaluation and pharmaceutics reformation of pueraria for intranasal.

Bioassay-guided fractionation led to the isolation of eight compounds from Stemona sessilifolia. Of the eight isolates, three new bibenzyls, stilbostemins M-O (1-3), and a new tocopherol, 6-methoxy-3,4-dehydro-delta-tocopherol (4) were revealed together with four known compounds 3,5-dihydroxy-2'-methoxy bibenzyl (5), 3,5-dihydroxy bibenzyl (6), beta-tocopherol (7), and gamma-tocopherol (8). Compounds 5, 6, and 8 exhibited strong antibacterial activities against Staphylococcus aureus and S. epidermidis.

We have explored the effect of photodynamic therapy (PDT) with verteporfin on the induction and expression of contact hypersensitivity (CHS) to 2,4-dinitrofluorobenzene (DNFB) in normal mice and IL-10-deficient mice. Our results indicate that DNFB sensitized mice given PDT with verteporfin and whole body red light irradiation exhibited a significant reduction in CHS compared with control animals. Administration of rIL-12 reversed the effect(s) of PDT as did treatment of mice with anti-IL-10-neutralizing Ab. Knockout mice deficient in IL-10 were found to be resistant to the inhibitory effects of PDT. In vitro proliferative responses using spleen cells from DNFB-sensitized and PDT-treated mice showed a significantly lower response to DNBS as compared with cells from DNFB-sensitized mice or DNFB and PDT-treated IL-10-deficient mice. Finally, naive mice exposed to PDT exhibited an increase in skin IL-10 levels, which peaked between 72 and 120 h post-PDT. Together these data support the role of IL-10 as a key modulator in the inhibition of the CHS response by whole body PDT.

Phosphatidylinositol 3-kinase (PI 3-kinase) and protein kinase B are critical players in cell proliferation and survival. Their downstream effector protein kinase, p70 S6 kinase, has an established role in protein translation. The mechanism by which bacterial LPS induces production of nitric oxide (NO) in murine macrophages is incompletely understood, and a role for PI 3-kinase/p70 S6 kinase pathway had not been previously investigated. In this study we demonstrate that LPS induced a fivefold activation of p70 S6 kinase and a twofold stimulation of PI 3-kinase. Pretreatment of Raw 264.7 cells with either rapamycin or Ly290042 completely blocked LPS-induced activation of p70 S6 kinase. Protein kinase B was also activated (twofold) by LPS and was only minimally affected by these inhibitors. PI 3-kinase activity was inhibited by both Ly294002 and wortmannin. The effects on NO production by these agents were strikingly different. While both rapamycin and Ly294002 resulted in almost complete inhibition of NO production, wortmannin was ineffective. Surprisingly, none of the inhibitors reduced the production of the inducible nitric oxide synthase protein (iNOS) as determined by immunoprecipitation. In vivo labeling studies revealed that the iNOS protein was phosphorylated in concordance with the production of NO. We conclude that LPS-mediated NO production occurs via a PI 3-kinase-independent, but FKBP12-rapamycin-associated protein-dependent, pathway in RAW cells by a mechanism probably involving phosphorylation of iNOS.

A quinol oxidase has been purified from the cytoplasmic membrane of Paracoccus denitrificans; its heme composition and CO binding properties identify it as a cytochrome ba3. On SDS gels, the purified enzyme complex is separated into five polypeptides. Using partial peptide sequence information for subunit II, the gene locus has been cloned and sequenced. In a typical operon pattern, four genes were identified: qoxA, -B, -C, and -D, coding for subunits II, I, III, and IV. DNA-derived amino acid sequence comparisons reveal extensive similarities to other members of the terminal oxidase superfamily.

We have constructed an expression vector for the phytopathogenic fungus Ustilago maydis. This vector, pUXV, expresses genes located downstream from a U. maydis glyceraldehyde-3-phosphate dehydrogenase promoter. Plasmid pUXV also contains a selective marker gene conferring resistance to the antibiotic hygromycin B and a U. maydis autonomously replicating sequence, UARS, allowing high transformation efficiency. Expression of a cDNA from the toxin-encoding region of the U. maydis virus P6 in pUXV resulted in as much killing activity as from viral particles when evaluated by killer plate assay. Plasmid pUXV preserves essential sequences from pUC12 and is therefore a shuttle vector for U. maydis and Escherichia coli.

The nucleotide sequence of the murE gene encoding the diaminopimelic acid adding enzyme of Escherichia coli is reported. The coding region consisted of 1413 base pairs and was separated from the ftsI (penicillin-binding protein 3) gene by 61 base pairs. The deduced primary structure of MurE comprised 471 amino acid residues with a molecular mass of 50.6 kilodaltons.