C-H-activated Csp2-Csp3 diastereoselective gridization enables ultraviolet-emitting stereo-molecular nanohydrocarbons with mulitple H···H interactions.

Gridization is an emerging molecular integration technology that enables the creation of multifunctional organic semiconductors through precise linkages. While Friedel-Crafts gridization of fluorenols is potent, direct linkage among fluorene molecules poses a challenge. Herein, we report an achiral Pd-PPh3-cataylized diastereoselective (>99:1 d.r.) gridization based on the C-H-activation of fluorene to give dimeric and trimeric windmill-type nanogrids (DWGs and TWGs). These non-conjugated stereo-nanogrids showcase intramolecular multiple H…H interactions with a low field shift to 8.51 ppm and circularly polarized luminescence with high luminescent dissymmetry factors (|gPL | = 0.012). Significantly, the nondoped organic light-emitting diodes (OLEDs) utilizing cis-trans-TWG1 emitter present an ultraviolet electroluminescent peak at ~386 nm (CIE: 0.17, 0.04) with a maximum external quantum efficiency of 4.17%, marking the highest record among nondoped ultraviolet OLEDs based on hydrocarbon compounds and the pioneering ultraviolet OLEDs based on macrocycles. These nanohydrocarbon offer potential nanoscafflolds for ultraviolet light-emitting optoelectronic applications.

U-shaped kirschner wire transfixation: effective treatment for Skier's thumb.

BACKGROUND: Skier's thumb is a type of injury to the ulnar collateral ligament of the metacarpophalangeal joint of the thumb, which can result in bone fragmentation and joint instability. OBJECTIVE: The objective of this study was to compare the traditional Kirschner wire fixation method with the U-shaped Kirschner wire method for treating small bone fragments with displacement, rotation, or instability in skier's fractures. METHOD: A retrospective study was conducted on 30 patients with skier's thumb who were treated at Tianjin Hospital from January 2019 to December 2021. Patients were divided into two groups: Group A received traditional Kirschner wire fixation, while Group B received U-shaped Kirschner wire fixation. Functional assessments and complications during the perioperative period were evaluated. RESULTS: Both surgical methods significantly reduced postoperative pain and increased joint range of motion. Group B had a lower incidence of pain during follow-up and showed significant functional improvement in Tip-pinch and Grip tests compared to Group A. U-shaped Kirschner wire fixation significantly reduced complications during the perioperative period. CONCLUSION: The U-shaped Kirschner wire internal fixation is a safe and effective treatment for the thumb proximal phalanx base ulnar side avulsion fracture.

A cuproptosis-related lncRNA signature for predicting prognosis and immune response in hepatocellular carcinoma.

Background: Hepatocellular carcinoma (HCC) has a high incidence and poor prognosis. Cuproptosis is a novel type of cell death, which differs from previously reported types of cell death such as apoptosis, autophagy, proptosis, ferroptosis, necroptosis, etc. Long non-coding RNAs (lncRNAs) play multiple roles in HCC. Methods: We downloaded information from The Cancer Genome Atlas (TCGA) database, and obtained cuproptosis-related genes from published studies. The cuproptosis-related lncRNAs were obtained by correlation analysis, and subsequently used to construct a prognostic cuproptosis-related lncRNA signature. Analyses of overall survival (OS), progression-free survival (PFS), receiver operating characteristic (ROC) curve with the area under the curve (AUC) values and the index of concordance (c-index) curve were used to evaluate the signature. The tumor microenvironment (TME) was analyzed by ESTIMATE algorithm. The immune cell data was downloaded from the Tumor Immune Estimation Resource (TIMER) 2.0 database. Immune-related pathways were analyzed by single-sample gene set enrichment analysis (ssGSEA) algorithm. Immunophenoscore (IPS) scores from The Cancer Immunome (TCIA) database were used to evaluate immunotherapy response. The "pRRophetic" was employed to screen drugs for high-risk patients. The candidate lncRNA expression levels were detected by Real Time Quantitative PCR. Results: We constructed a cuproptosis-related lncRNA signature containing seven lncRNAs: AC125437.1, PCED1B-AS1, PICSAR, AP001372.2, AC027097.1, LINC00479, and SLC6A1-AS1. This signature had excellent accuracy, and was independent of the stratification of clinicopathological features. Further study showed that high-risk tumors under this signature had higher TMB, fewer TME components and higher tumor purity. The tumors with high risk were not enriched in immune cell infiltration or immune process pathways, and high-risk patients had a poor response to immunotherapy. Moreover, 29 drugs such as sorafenib, dasatinib and paclitaxel were screened for high-risk HCC patients to improve their prognosis. The expression levels of the candidate lncRNAs in HCC tissue were significantly increased (except PCED1B-AS1). Conclusions: Our prognostic cuproptosis-related lncRNA signature was accurate and effective for predicting the prognosis of HCC. The immunotherapy was unsuitable for high-risk HCC patients with this signature.

Bioinformatic Analysis of Key Regulatory Genes in Adult Asthma and Prediction of Potential Drug Candidates.

Asthma is a common chronic disease that is characterized by respiratory symptoms including cough, wheeze, shortness of breath, and chest tightness. The underlying mechanisms of this disease are not fully elucidated, so more research is needed to identify better therapeutic compounds and biomarkers to improve disease outcomes. In this present study, we used bioinformatics to analyze the gene expression of adult asthma in publicly available microarray datasets to identify putative therapeutic molecules for this disease. We first compared gene expression in healthy volunteers and adult asthma patients to obtain differentially expressed genes (DEGs) for further analysis. A final gene expression signature of 49 genes, including 34 upregulated and 15 downregulated genes, was obtained. Protein-protein interaction and hub analyses showed that 10 genes, including POSTN, CPA3, CCL26, SERPINB2, CLCA1, TPSAB1, TPSB2, MUC5B, BPIFA1, and CST1, may be hub genes. Then, the L1000CDS2 search engine was used for drug repurposing studies. The top approved drug candidate predicted to reverse the asthma gene signature was lovastatin. Clustergram results showed that lovastatin may perturb MUC5B expression. Moreover, molecular docking, molecular dynamics simulation, and computational alanine scanning results supported the notion that lovastatin may interact with MUC5B via key residues such as Thr80, Thr91, Leu93, and Gln105. In summary, by analyzing gene expression signatures, hub genes, and therapeutic perturbation, we show that lovastatin is an approved drug candidate that may have potential for treating adult asthma.

Optimization of production system of shale oil development in Ordos basin, China.

In this paper, production system (PS) of shale oil is optimized according to production data and indoor experiments, including core and fluid tests. Results showed that: ① Pressure drop rate at wellhead is a reasonable reference for the determination of post-fracture shut-in duration (PFSID). When pressure at a wellhead of horizontal well is relatively stable and the pressure drop less than 0.1 MPa per day for three consecutive days, PFSID ends; ② Flowback intensity of fracturing fluid affects the effectiveness of proppant underground, thus flowback intensity can be determined by the critical flow rate and safety factor of each proppant; ③ Flowback intensity should be varied during different development stages, which could be divided into four according to production gas and oil ratio(GOR) of a shale oil horizontal well: low, medium-high, high and high-low production GOR. During the stage of low production GOR, ratio of flow pressure and saturation pressure should be maintained greater than 1.0, and the initial daily liquid productivity for a hundred-meter oil-bearing lateral length in a horizontal well is 2.4 ~ 2.9 m3/d; and during the medium-high production GOR, high production GOR and high-low production GOR stages, the responding initial daily liquid productivity should be maintained between 0.8 ~ 1.0 or less than 0.8 respectively.

An Eco-Friendly Acid Leaching Strategy for Dealkalization of Red Mud by Controlling Phase Transformation.

The alkaline components in red mud represent one of the crucial factors restricting its application, especially for the construction and building industry. The phase state of alkaline components has a significant influence on the dealkalization of red mud. In this work, an environmentally friendly acid leaching strategy is proposed by controlling the phase transformation of red mud during active roasting pretreatment. With a moderate roasting temperature, the alkaline component is prevented from converting into insoluble phases. After acid leaching with a low concentration of 0.1 M, a high dealkalization rate of 92.8% is obtained. Besides, the leachate is neutral (pH = 7) and the valuable metals in red mud are well preserved, manifesting a high selectivity and efficiency of diluted acid leaching. The calcination experiment further confirms the practicability of the strategy in the construction field, where the cementitious minerals can be formed in large quantities. Compared with the traditional acid leaching routes, the diluted acid leaching strategy in this work is acid saving with low valuable element consumption. Meanwhile, the secondary pollution issue can be alleviated. Hence, the findings in this work provide a feasible approach for the separation and recovery of alkali and resource utilization of red mud.

Effects of replacing fishmeal with cottonseed protein concentrate on growth performance, blood metabolites, and the intestinal health of juvenile rainbow trout (Oncorhynchus mykiss).

Cottonseed protein concentrate (CPC) is a potential non-food protein source for fishmeal replacement in fish feed. However, a high inclusion level of CPC in diets may have adverse effects on the metabolism and health of carnivorous fish. This study aimed to investigate CPC as a fishmeal alternative in the diet of rainbow trout Oncorhynchus mykiss based on growth performance, blood metabolites, and intestinal health. Five isonitrogenous (46% crude protein) and isolipidic (16% crude lipid) diets were formulated: a control diet (30% fishmeal) and four experimental diets with substitution of fishmeal by CPC at 25%, 50%, 75%, and 100%. A total of 600 fish (mean body weight 11.24g) were hand-fed the five formulated diets to apparent satiation for eight weeks. The results showed no adverse effects on growth performance when 75% dietary fishmeal was replaced by CPC. However, reduced growth and feed intake were observed in rainbow trout fed a fishmeal-free diet based on CPC (CPC100%). Changes in serum metabolites were also observed in CPC100% compared with the control group, including an increase in alanine aminotransferase (ALT), a decrease in alkaline phosphatase (ALP), alterations in free amino acids, and reductions in cholesterol metabolism. In addition, the CPC-based diet resulted in reduced intestinal trypsin, decreased villus height and width in the distal intestine, upregulated mRNA expression levels of inflammatory cytokines in the intestine, and impaired gut microbiota with reduced bacterial diversity and decreased abundance of Bacillaceae compared with the control group. The findings suggest that the optimum substitution rate of dietary fishmeal by CPC for rainbow trout should be less than 75%.

Altered circular RNA expression profiles in an ovalbumin-induced murine model of allergic rhinitis

Background Emerging evidence shows that circular RNAs (circRNAs) participate in the pathogenesis of multiple immune diseases. However, few studies have focused on the mechanisms of circRNAs involved in allergic rhinitis (AR). Methods This study performed an RNA sequence (RNA-seq) profiling to identify the expression of circRNAs in nasal mucosa from ovalbumin-induced AR murine models and normal controls. Quantitative real-time reverse transcriptase polymerase chain reaction (qRT-PCR) was then conducted to validate the differential expression of circRNAs. Bioinformatics analysis was applied to demonstrate the biological functions of the dysregulated circRNAs. Results A total of 86 distinct circRNA candidates were sequenced, of which 51 were upregulated and 35 were downregulated. The T cell receptor, B cell receptor, and calcium signaling pathways may be involved in the pathology of AR. Furthermore, a circRNA-miRNA interaction network was constructed via miRNA response elements analysis. Some circRNAs were correlated with miRNAs that are involved in T cell polarization and activation, thereby highlighting their potential role in the pathogenesis of AR. Conclusions This study demonstrates a number of aberrantly expressed circRNAs related to AR, and offers a novel perspective into AR pathogenesis and future therapeutic strategies (AU)

Altered circular RNA expression profiles in an ovalbumin-induced murine model of allergic rhinitis.

BACKGROUND: Emerging evidence shows that circular RNAs (circRNAs) participate in the pathogenesis of multiple immune diseases. However, few studies have focused on the mechanisms of circRNAs involved in allergic rhinitis (AR). METHODS: This study performed an RNA sequence (RNA-seq) profiling to identify the expression of circRNAs in nasal mucosa from ovalbumin-induced AR murine models and normal controls. Quantitative real-time reverse transcriptase polymerase chain reaction (qRT-PCR) was then conducted to validate the differential expression of circRNAs. Bioinformatics analysis was applied to demonstrate the biological functions of the dysregulated circRNAs. RESULTS: A total of 86 distinct circRNA candidates were sequenced, of which 51 were upregulated and 35 were downregulated. The T cell receptor, B cell receptor, and calcium signaling pathways may be involved in the pathology of AR. Furthermore, a circRNA-miRNA interaction network was constructed via miRNA response elements analysis. Some circRNAs were correlated with miRNAs that are involved in T cell polarization and activation, thereby highlighting their potential role in the pathogenesis of AR. CONCLUSIONS: This study demonstrates a number of aberrantly expressed circRNAs related to AR, and offers a novel perspective into AR pathogenesis and future therapeutic strategies.

[Comparison and intervention of differences in upper airway obstruction in children with OSAS between awake and asleep].

Objective:Drug induced sleep endoscopy（DISE） is a useful tool to locate the upper airway obstructions in patients with obstructive sleep apnea syndrome（OSAS）. The goal of our research was to compare the sites, degree and patterns of upper airway obstruction detected by DISE versus endoscopy in awake children. The benefit of DISE-mediated upper airway surgery in pediatric OSAS was evaluated. Method:This is a retrospective case series of children with OSAS who had adenoid hypertrophy（the area where adenoids block the postnaris ≥2/3） and tonsils ≤2 degrees（Friedman classification method） . All children underwent sleep monitoring by pulse oximeter before and after operation. DISE was performed before adenoidectomy in DISE group. If obstruction of oropharynx caused by tonsils was found, tonsillectomy was performed at the same time with adenoidectomy. The children who did not undergo DISE before operation were enrolled in the control group. Differences in the degrees and patterns of upper airway obstruction in the DISE group under wakefulness and drug induction were analyzed. Result:The study included 126 children in DISE group, and 200 children in control group. In the DISE group, 56 cases（44.4%） had grade 2 tonsils, and 70 cases（55.6%） had grade 1 tonsils. The patterns of obstruction at velum, tongue base and epiglottis were significantly different in DISE compared with awake endoscopy（P<0.05）; the patterns of obstruction at adenoid and oropharynx were the same in both techniques. There were significant differences in endoscopy scores between the lateral wall of oropharynx and the tongue base（P<0.05）. DISE revealed 57 （45.2%） cases had grade 2 obstruction in the oropharyngeal level. Among them, 44 cases （34.9%） had tonsils at 2 degrees and 13 cases （10.3%） had tonsils at 1 degrees.. There was an improvement in respiratory disturbance index（RDI） and oxygen saturation nadir in the DISE and control groups 1 year after surgery（P<0.05）, and the RDI improvement was better in the DISE group compared with controls（P=0.04）. Conclusion:DISE is an effective method to evaluate the sites and severity of upper airway obstruction in children with OSAS, which was conductive to make the upper airway personalized surgical plan for patients.

High-Performance Pr3+-Doped Scandate Optical Thermometry: 200 K of Sensing Range with Relative Temperature Sensitivity above 2%·K-1.

Nowadays, for lanthanide fluorescence thermometry, high relative temperature sensitivity (Sr) and wide sensing range are urgently required in practical applications. Herein, a Pr3+-doped scandate (Pr3+:CaSc2O4) luminescent thermometer is proposed, and the related crystal-field splitting of Pr3+ ions is systematically discussed. The 4f5d-4f and 4f-4f emissions of Pr3+ ions in CaSc2O4 basically present positive and negative correlation relationships with the increase in temperature, respectively. The different changing tendencies in relation to temperature for the two kinds of transitions are mainly derived from the effects of the thermally activated trap energy levels and the crossover process between 5d and 4f energy states of Pr3+ ions and are beneficial for the enhancement of relevant temperature sensitivities. The obtained experimental results manifest that Pr3+:CaSc2O4 owns a maximum relative temperature sensitivity Sr of 2.49%·K-1 (at 390 K) and a low temperature uncertainty (around 0.1 K from 275 to 490 K). Moreover, it is also able to keep a relatively high Sr (not lower than 2%·K-1) over a wide temperature sensing range (∼200 K), which is more excellent than those of reported luminescent thermometric materials (∼100 K). Hence, what discussed in this study might provide a useful design perspective for the exploration and development of high-performance luminescent thermometers with a wide applicable temperature range.

Chemical characterization of polysaccharides isolated from scrophularia ningpoensis and its protective effect on the cerebral ischemia/reperfusin injury in rat model.

This study was designed to investigate the chemical characterization of scrophularia ningpoensis polysaccharides (SNP) and its protective activity on the cerebral ischemia/reperfusin injury (CIRI) rats. SNP was successfully isolated and fractionated by anion exchange chromatography and gel filtration into different factions, namely SNP1, SNP2, SNP3, SNP2-A and SNP2-B. SNP2-A is consisted of mannose (Man), rhamnose (Rha), glucuronic Acid (Glu A), galacturonic Acid (Gal A), glucose (Glu), galactose (Gal), xylose (Xyl) and arabinose (Ara). The nervous defect symptoms, the brain infarct volume and brain water content of rats with CIRI were improved by pretreatment with SNP2-A. In addition, SNP2-A could increase SOD activities, but could reduce MDA, NO, NOS, IL-1ß and TNF-α levels in brain tissues of rats with CIRI. Additionally, the reductive ERK protein expression and the elevated JNK and p38 protein expression stimulated by CIRI were all ameliorated by pre-treatment of SNP2-A. The results indicate that the protective effect may be related to the improvement of SNP2-A on the antioxidant capacity of brain tissue, the inhibition of inflammatory cytokines overproduction and the amelioration of MAPK pathway such as JNK, p38 and ERK proteins. These data provided an information for mechanistic studies for polysaccharide neuroprotective effects in CIRI.

Development of homologous enzyme-linked immunosorbent assays to quantify two forms of vitellogenin in guppy (Poecilia reticulata).

Guppy (Poecilia reticulata) is a promising model organism in toxicological studies, and vitellogenin (Vtg) is a commonly used biomarker for environmental estrogens. Although an ELISA for guppy Vtg has been developed previously, we found that guppy had two forms of Vtgs. In this study, two Vtgs were characterized and enzyme-linked immunosorbent assays (ELISAs) for each Vtg were developed. Two Vtgs purified from 17ß-estradiol (E2)-exposed guppy were characterized as phospholipoglycoproteins with molecular weights of ~ 520 and ~ 480 kDa, respectively. In SDS-PAGE, one purified Vtg appeared as three major bands of ~ 210, ~ 126, and ~ 102 kDa, and the other revealed a clear band of ~ 68 kDa. Matrix-assisted laser desorption/ionization-time of flight/time of flight mass spectrometry analysis showed that they were VtgAb and VtgC. Using purified Vtgs and their corresponding antibodies, two sandwich ELISAs with working ranges of 7.8~1000 and 15.6~500 ng/mL were developed. Precision tests showed that intra- and inter-assay coefficients of variations of both ELISAs were below 10%. Parallelism between Vtg standard curves and serial dilutions of whole body homogenate from E2-exposed guppy confirmed that two ELISAs could quantify guppy Vtgs. Furthermore, two ELISAs were used to measure Vtg inductions in liver, caudal fin and whole body of male guppy exposed to 17a-ethinylestradiol to validate their use for detecting estrogenic effects of exogenous chemicals. These homologous Vtg ELISAs will promote the use of guppy as a model organism to study estrogenic chemicals.

[Optimize preparation of compound licorice microemulsion with D-optimal design].

In order to increase the solubility of essential oil in compound licorice microemulsion and improve the efficacy of the decoction for treating chronic eczema, this experiment intends to prepare the decoction into microemulsion. The essential oil was used as the oil phase of the microemulsion and the extract was used as the water phase. Then the microemulsion area and maximum ratio of water capacity was obtained by plotting pseudo-ternary phase diagram, to determine the appropriate types of surfactant and cosurfactant, and Km value-the mass ratio between surfactant and cosurfactant. With particle size and skin retention of active ingredients as the index, microemulsion prescription was optimized by D-optimal design method, to investigate the in vitro release behavior of the optimized prescription. The results showed that the microemulsion was optimal with tween-80 as the surfactant and anhydrous ethanol as the cosurfactant. When the Km value was 1, the area of the microemulsion region was largest while when the concentration of extract was 0.5 g·mL⁻¹, it had lowest effect on the particle size distribution of microemulsion. The final optimized formulation was as follows: 9.4% tween-80, 9.4% anhydrous ethanol, 1.0% peppermint oil and 80.2% 0.5 g·mL⁻¹ extract. The microemulsion prepared under these conditions had a small viscosity, good stability and high skin retention of drug; in vitro release experiment showed that microemulsion had a sustained-release effect on glycyrrhizic acid and liquiritin, basically achieving the expected purpose of the project.

Transdermal Delivery of Compounds with Different Lipophilicity and Molecular Weight from W/O Microemulsions Analyzed by UPLC-QTOF/ MS and LC-MS/MS.

OBJECTIVE: The aim of this study was to develop a novel W/O microemulsion for a natural extract of Wen-Luo-Tong (WLT) containing mainly icariin, hydroxysafflor yellow A (HSYA) and gallic acid to be applied to skin as a potential treatment for peripheral neuropathy. METHODS: The oil phase was selected on the basis of affinity with the surfactant and co-surfactant. Pseudo-ternary diagrams were constructed to optimize microemulsions and finally stability studies were performed on the selected formulations. Droplet sizes were analyzed by using a zetasizer and were found to be within the desired range. Selected microemulsions with acceptable viscosities, containing 5%, 8% and 10% of water extract solution, were used for in vitro skin penetration studies using Franz diffusion cells and excised rat skin. New LC-MS/MS and UPLC-Q-TOF/MS methods were employed for quantitative and qualitative analysis. RESULTS: The optimized formulation (ME-4) consisting of 10% (w/w) water extract solution, 60% isopropyl myristate, 30%(w/w) Smix: Propylene glycol (5:2) significantly increased the cumulative permeated amounts of HSYA, icariin and gallic acid compared with the water extract solution controls. CONCLUSION: This novel formulation also increased the number of components penetrating rat skin. Ten components were detected in the Franz cell receptor solution using a UPLC-Q-TOF/MS system after the application of formulation ME-4 for 24h on the skin in vitro. However, only one component was detected after applying the control. Therefore, the microemulsion ME-4 was selected for future in vivo pharmacodynamic studies.

Vitellogenin induction in caudal fin of guppy (Poecilia reticulata) as a less invasive and sensitive biomarker for environmental estrogens.

Guppy (Poecilia reticulata) is an ideal model for studying environmental estrogens, and its large caudal fin has a high capacity to regenerate. This study analyzed the feasibility of caudal fin for detecting vitellogenin (Vtg), the most commonly used biomarker of environmental estrogens. Firstly, a sandwich ELISA for guppy Vtg was developed using purified lipovitellin and its antibody and it had a working range of 7.8-1000 ng/mL and detection limit of 3.1 ng/mL. The ELISA was used to detect tissue distribution of Vtg. In male guppy exposed to 50 and 100 ng/L 17ß-estradiol (E2), Vtg concentration in caudal fin was higher than that in whole fish, brain, eyes, gonad, and skin, and was close to that in the liver. Furthermore, male guppies were exposed to environmental concentrations of 17a-ethinylestradiol (EE2) and bisphenol S (BPS) to validate the utility of caudal fin Vtg for detecting estrogenic activities. The lowest observed effect concentration of EE2 and BPS were lower than 2 ng/L and 1 µg/L, which were below or equal to the values reported for other species, demonstrating that caudal fin Vtg was highly sensitive to estrogenic chemicals. Therefore, caudal fins of guppies are suggested as alternative samples for Vtg biomarker detection.

Lipovitellin as an antigen to improve the precision of sandwich ELISA for quantifying zebrafish (Danio rerio) vitellogenin.

Vitellogenin (Vtg) in zebrafish (Danio rerio) is a core biomarker for screening environmental estrogens in test guidelines of the Organization for Economic Cooperation and Development. To accurately quantify zebrafish Vtg, lipovitellin (Lv), the main Vtg-derived yolk protein, was used as the antigen to establish a sandwich enzyme-linked immunosorbent assay (ELISA). The purified Lv was a phospholipoglycoprotein with apparent molecular weight of ~445kDa, and separated into three polypeptides corresponding to ~117, ~102, and ~23.8kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Immunological analysis confirmed the specificity of the anti-Lv antibody for Vtg and the immunological similarity between Vtg and Lv. Using the purified Lv and anti-Lv antibody, a sandwich ELISA with a detection limit of 4.3ng/mL and a detection range from 7.8 to 250ng/mL was developed. The intra- and inter-assay coefficients of variation were both below 10%. Moreover, the Lv standard curve was nearly identical to the Vtg standard curve, and paralleled serial whole-body homogenate dilutions of male zebrafish exposed to 17ß-estradiol, demonstrating that the Lv-based ELISA could be used for quantification of zebrafish Vtg. Zebrafish Lv showed high stability during purification process, heat treatment, -80°C storage, and repeated freeze/thaw cycles. Additionally, the standard curve of Lv stored at -80°C for 3months exhibited higher robustness than that of Vtg stored under the same conditions. Finally, the usefulness of the ELISA for detecting estrogenic activity was verified by quantifying Vtg inductions in zebrafish exposed to monocrotophos.

[Optimize preparation parameters of colon-specific pellets of Angelica Sinensis Radix supercritical fluid extraction with response surface analysis methodology].

To prepare pellets of supercritical fluid extraction (SFE) of Angelica Sinensis Radix by using the ionic crosslinking method, and the drug loading and encapsulation efficiency were used as the index to investigate the multiple factors which may impact the drug loading and encapsulation efficiency. Box-Behnken design and response surface analysis method were then taken to optimize the prescription of pellets and study the coating technology. Through the study on the release of pellets in vitro, an optimal coating technology and prescription of colon-specific pellets of Angelica Sinensis Radix SFE were selected and their colon targeting was evaluated. The optimal preparation parameters of pellets were determined as follows： 3% pectin; 4∶1 for pectin/lecithin; 4∶5 for pectin/SFE of Angelica Sinensis Radix; 4% zinc acetate solution as crosslinking agent, blending temperature 35 ℃, crosslinking temperature 35 ℃, crosslinking time 30 min; coating technology： coating material Eudragit FS 30D, 1.5% triethyl citrate and polyoxyethylene sorbitan monooleate（tween-80）, 1.2% monostearin and 15% coating weight gained. The colon-specific pellets of Angelica Sinensis Radix SFE prepared with optimized conditions were almost not released in simulated gastric fluid in 2 h, released less than 20% in simulated intestine fluid in 4 h, and released more than 90% in simulated colon fluid in 6 h, indicating that the colon-specific pellets of Angelica Sinensis Radix SFE had an excellent colon targeting property.

Transport of ginkgolides with different lipophilicities based on an hCMEC/D3 cell monolayer as a blood-brain barrier cell model.

AIMS: In this report, the transport of ginkgolides with different lipophilicities was investigated using an hCMEC/D3 cell monolayer as a blood-brain barrier (BBB) cell model in vitro in an attempt to explain ginkgolide transport path mediated by lipophilicity. MAIN METHODS: The log P values of ginkgolides were determined by measuring the distribution of the molecule between oil and water. Additionally, the cytotoxicity of ginkgolides on hCMEC/D3 cells was assayed with the MTT method. Ginkgolide contents were determined with an ultra performance liquid chromatograph equipped with an evaporative light scattering detector (ULPC-ELSD) method. Apparent permeability coefficients (Papp) and efflux ratios (PappBL→AP/PappAP→BL) were then calculated to describe the transport characteristics of ginkgolide. KEY FINDINGS: The transport of ginkgolide A, ginkgolide B, ginkgolide C, and ginkgolide J across the hCMEC/D3 cell monolayer was non-directional. Additionally, ginkgolide C transport on the cell monolayer was time- and concentration-dependent in the paracellular pathway controlled by cytochalasin D (a tight junction modulator). The transport of ginkgolide N, ginkgolide L, and ginkgolide K across the cell monolayer displayed clear directionality at low ginkgolide concentrations. This behavior indicated that the transport of ginkgolide N, ginkgolide L, and ginkgolide K was influenced by the transcellular pathway containing an efflux protein accompanied by the paracellular pathway for passive diffusion. Additionally, the transport of ginkgolide K was increased significantly by co-culturing with a P-gp inhibitor. SIGNIFICANCE: These findings provide important information for elucidating ginkgolide transport pathways and may be beneficial for the design of ginkgolide molecules with high neuroprotective effects.

Neuroprotective effect of Ginkgolide K against H2O2-induced PC12 cell cytotoxicity by ameliorating mitochondrial dysfunction and oxidative stress.

Mitochondria and oxidative stress play important roles in neuronal cell death associated with cerebral ischemia. Elevated level of reactive oxygen species (ROS) and mitochondrial dysfunction are thought to be responsible for cerebral ischemia injury along with neural cells death through several apoptotic mechanisms. In this study, exposure of rat pheochromocytoma (PC12) cells to hydrogen peroxide (H2O2) at the concentration of 0.3 mM for 24 h caused significant loss of cell viability, lactate dehydrogenase (LDH) release from cells, ascent of ROS level and mitochondrial membrane potential (MMP) decrease. Moreover, the activities of caspase-9, caspase-8 and caspase-3 all were increased in H2O2-induced PC12 cells. However, pretreatment with ginkgolide K (GK) solutions of different concentrations (10, 50, 100 µM) for 24 h prior to exposuring to H2O2 significantly increased cells viability, suppressed LDH release, attenuated ROS level, prevented cytochrome c release from mitochondria and boosted MMP expression. In addition, ginkgolide K notably inhibited the caspase-3 and caspase-9 but not caspase-8 activities in exogenous H2O2-treated PC12 cells. These results demonstrated that ginkgolide K protected PC12 cells from H2O2-induced apoptosis by restoring MMP expression, ameliorating oxidative stress and subsequently leading to inhibit the activity of caspase-3 protein. Therefore, the present study supported that ginkgolide K may be a promising neuroprotective compound for cerebral ischemia treatment.