Blood-supplementing effect of low molecular weight peptides of E-Jiao on chemotherapy-induced myelosuppression: evaluation of pharmacological activity and identification of bioactive peptides released in vivo.

Background: Equus asinus L. [Equidae; Asini Corri Colla] (donkey-hide gelatin, E-Jiao) is a traditional Chinese medicine renowned for its exceptional blood-supplementing effect. However, the specific components that contribute to its efficacy remain elusive. This study aimed to demonstrate that peptides are responsible for E-Jiao's blood-supplementing effect and to explore the specific peptides contributing to its efficacy. Methods: The low molecular weight peptides of E-Jiao (LMEJ) were obtained using an in vitro digestion method. LMEJ and peptides in the rat bloodstream were characterized by peptidomics analysis. The blood-supplementing effect of LMEJ was assessed using blood-deficient zebrafish and mouse models. The effect of the peptides detected in rat blood was evaluated using the same zebrafish model, and network pharmacology analysis was performed to investigate the underlying mechanisms. Results: A total of 660 unique peptides were identified within LMEJ. Both E-Jiao and LMEJ significantly alleviated myelosuppression in mice but only LMEJ attenuated myelosuppression in zebrafish. After the administration of E-Jiao to rats, 67 E-Jiao-derived peptides were detected in the bloodstream, 41 of which were identical to those identified in LMEJ. Out of these 41 peptides, five were synthesized. Subsequent verification of their effects revealed that two of them were able to alleviate myelosuppression in zebrafish. Network pharmacology study suggested that E-Jiao may exert a blood-supplementing effect by regulating signaling pathways such as JAK-STAT, IL-17 and others. These results indicated that peptides are at least partially responsible for E-Jiao's efficacy. Conclusion: This study provides a crucial foundation for further exploration of the bioactive components of E-Jiao.

Analytical strategies to identify multicomponent quality markers for commercial Hua-ju-hong using multidimensional chemical analysis.

Hua-ju-hong (HJH) is a Chinese medicinal material obtained from Citrus grandis 'Tomentosa' (CGT) and Citrus grandis (L.) Osbeck (CG) with various commercial specifications. It is known for relieving cough and dispelling phlegm. To reveal the quality marker for distinguishing the various HJH, 215 batches of commercial HJH were studied systematically using multidimensional chemical analysis. Ten major components were identified by high-performance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry and quantified via high-performance liquid chromatography coupled with diode array detection. In this study, a rapid, efficient, and low-cost chromatographic method was established. Total coumarin-hemiterpene and total coumarin-monoterpene were first classified and analyzed in HJH. The result indicated that the main component, naringin, was not the quality marker for differentiating CGT from CG. For reflecting the unique medicinal and food value of HJH, coumarins should be the more potential quality markers. Flavonoids were the possible quality markers for distinguishing two growth stages of fruit-exocarp and young fruit. For the first time, two chemotypes of HJH were identified in CG. This study provides a convenient yet reliant chromatographic method and novel yet systematic strategies for overall quality control of commercial HJH.

Is exponential stability achievable in singular perturbed delayed systems with time-varying parameters? A comprehensive analysis.

The present article conducts an investigation into the phenomenon of exponential stability within singular perturbed delayed systems, incorporating time-varying parameters. Singularly perturbed systems serve as essential tools in modeling intricate systems characterized by multiple time scales, wherein one subsystem exhibits significantly faster evolution than the others. The presence of small delays introduces complexities, influencing both state derivatives and delays, further accentuating the intricacies of the system. Drawing upon the principles of singular perturbation theory, the article introduces a novel approach to analyzing the stability of these complex systems, eschewing the conventional assumption of exponential stability in the fast subsystem. Within the scope of this study, we propose a rigorous stability analysis, utilizing Linear Matrix Inequality (LMI) methods, while considering time-varying parameters that exert substantial influence on the system's dynamics. The proposed methodology enables the exploration of system stability beyond conventional assumptions, imparting valuable insights into the behavior of singular perturbed delayed systems amidst varying conditions. Through extensive numerical simulations, the effectiveness and robustness of the approach are validated, illuminating the stability properties of these intricate systems. Comparative studies with existing techniques, which assume exponential stability in the fast subsystem, demonstrate the distinct advantages and uniqueness of the presented approach. The findings underscore the significance of accounting for time-varying parameters in achieving a comprehensive understanding of the exponential stability inherent in singular perturbed delayed systems. This research makes substantial contributions to the field of system stability analysis, particularly in the context of singular perturbed delayed systems featuring time-varying parameters. The originality of our approach lies in introducing a comprehensive analysis framework that overcomes the limitations of existing methodologies. By integrating a novel stability analysis method based on Linear Matrix Inequalities (LMIs), we offer a fresh perspective on achieving exponential stability in such complex systems. Significantly, our work addresses a critical gap in current literature by challenging the assumption of exponential stability in the fast subsystem, a key feature of singularly perturbed systems. Through a meticulous examination of time-varying parameters, we unveil their profound impact on system dynamics, thus enriching the understanding of stability behaviors. The potential real-world applications of our findings span diverse fields, ranging from engineering to mathematical modeling. Performance metrics are a key focal point of our research. Numerical simulations employing our proposed LMIs serve as a robust benchmark, demonstrating the superior stability achieved in comparison to existing methods. This performance-driven evaluation ensures the practical applicability and reliability of our analysis approach across various scenarios.

Efficacy and safety of Qiangli Dingxuan tablet combined with amlodipine besylate for essential hypertension: a randomized, double-blind, placebo-controlled, parallel-group, multicenter trial.

Background: Hypertension, a major cardiovascular risk factor, severely impacts patients' quality of life. Qiangli Dingxuan tablet (QDT) is a formally approved Chinese patent medicine, which has been widely used as an adjunctive treatment for hypertension. This study aimed to investigate the antihypertensive efficacy and safety of QDT combined with amlodipine besylate in patients with essential hypertension. Methods: In this randomized, double-blind, placebo-controlled, parallel-group, multicenter trial conducted in China, patients diagnosed with grade 1 to 2 essential hypertension were randomly assigned in a 1:1 to the treatment of QDT or placebo for 12 weeks, alongside their ongoing treatment with amlodipine besylate. The primary outcome was the change in office blood pressure (BP) from baseline to 12 weeks. In addition, safety analysis included the assessment of vital signs and laboratory values. Results: At baseline, 269 patients were randomly assigned to the QDT group (n = 133) or the placebo group (n = 136), and there were no significant differences in baseline characteristics between the two groups. The primary outcome based on the full analysis set from baseline to 12 weeks showed that the mean difference in the change of office systolic BP reduction between the two groups was 6.86 mmHg (95%CI, 4.84 to 8.88, p < 0.0001), for office diastolic BP, the mean difference in the change of office diastolic BP reduction between the two groups was 4.64 mmHg (95%CI, 3.10 to 6.18, p < 0.0001). In addition, traditional Chinese medicine symptom scores were significantly decreased in the QDT group compared with the placebo group. No severe adverse events attributable to QDT were reported. Conclusion: The combination of QDT and amlodipine besylate demonstrates superior efficacy compared to amlodipine besylate monotherapy in the management of essential hypertension. QDT shows potential as an adjunctive treatment for essential hypertension. However, further rigorous clinical trials are warranted to validate these findings. Clinical Trial Registration: [https://clinicaltrials.gov/study/NCT05521282?cond=NCT05521282&rank=1]; Identifier: [NCT05521282].

Copper-Catalyzed Regio- and Stereoselective Sulfonylation of Alkynyl Imines with Sulfonyl Hydrazides: Access to (E)-ß-Sulfonyl Enones.

A copper-catalyzed sulfonylation of alkynyl imines with sulfonyl hydrazides has been developed, which exhibited excellent regio- and stereoselectivity and furnished a series of (E)-ß-sulfonyl enones in moderate to good yields. Mechanistic studies suggest that this strategy goes through a radical process.

TBAI-Catalyzed S-H and N-H Insertion Reactions of α-Diazoesters with Thiophenols and Amines under Metal-Free Conditions.

Mild, convenient, and effective TBAI-catalyzed S-H and N-H insertion reactions of α-diazoesters with thiophenols and aromatic amines under metal-free conditions have been described, furnishing a straightforward and general platform for the synthesis of various thioethers and 2-amino-2-oxoacetates in moderate to excellent yields. Moreover, this strategy features simple operation, mild conditions, broad substrate scope, and easy scale-up.

Species-specific identification of donkey-hide gelatin and its adulterants using marker peptides.

Donkey-hide gelatin is an important traditional Chinese medicine made from donkey skin. Despite decades of effort, identifying the animal materials (donkeys, horses, cattle and pigs) in donkey-hide gelatin remains challenging. In our study, we aimed to identify marker peptides of donkey-hide gelatin and its adulterants and develop a liquid chromatography-tandem mass spectrometry method to identify them. Theoretical marker peptides of four animals (donkeys, horses, cattle and pigs) were predicted and verified by proteomic experiments, and 12 species-specific marker peptides from donkey-hide gelatin and its adulterants were identified. One marker peptide for each gelatin was selected to develop the liquid chromatography-tandem mass spectrometry method. The applicability of the method was evaluated by investigating homemade mixed gelatin samples and commercial donkey-hide gelatin products. Using the liquid chromatography-tandem mass spectrometry method, the addition of cattle-hide gelatin and pig-hide gelatin to donkey-hide gelatin could be detected at a level of 0.1%. Horse-hide gelatin was detected when added at a level of 0.5%. Among 18 batches of donkey-hide gelatin products, nine were identified as authentic, and eight of the remaining samples were suspected to be adulterated with horse materials. These results provide both a practical method to control the quality of donkey-hide gelatin and a good reference for quality evaluations of other medicinal materials and foods containing protein components.

Molecular Identification of Commercial Fish Maws by DNA Sequencing of 16S rRNA and Cytochrome c Oxidase I Genes.

ABSTRACT: Fish maws (dried swim bladders) have long been used for medicinal tonics and as a valuable food resource in Southeast Asia. However, it is difficult to identify the original species of fish maws sold in markets due to a lack of taxonomic characteristics. In the present study, 37 kinds of commercial fish maws from various medicinal material markets were examined, and gene sequences were successfully obtained from ca. 95% of the samples. Partial sequences of the 16S rRNA gene and cytochrome c oxidase I (COI) gene were obtained and used to investigate the origin of these commercial fish maws. Thirty-five specimens belonged to nine species: five croakers and four noncroakers. All species identification was supported by both high homogeneity (98 to 100%) and clear clustering with low within-group Kimura two-parameter divergence scores (0 to 0.04 for 16S rRNA and 0 to 0.07 for COI) and high between-group divergence scores (0.07 to 0.15 for 16S rRNA and 0.11 to 0.24 for COI). Croakers were the predominant species, accounting for 74% of the total fish maw specimens. The large demand for croakers has put some species at the risk of extinction due to overfishing. As a valuable food, fish maw has progressively become more popular and has been used as a substitute for shark fin. The identification results allowed us to learn more about the fish species available on the fish maw market and provided an indicator for possible control of threatened or endangered fish species. A probable correlation between the molecular characteristics and morphological features of fish maws was also found and could provide both consumers and merchants with an important reference for identifying the origin of fish maws.

Comparisons of the anti-inflammatory, antiviral, and hemostatic activities and chemical profiles of raw and charred Schizonepetae Spica.

ETHNOPHARMACOLOGICAL RELEVANCE: A common view in traditional Chinese medicine (TCM) theory is that "processing can alter the efficacy of crude drugs". The clinical usage of some processed products may have already changed greatly over time during the development of modern scientific analysis. Therefore, the view of "processing can alter the efficacy of crude drugs" should be confirmed by comparative studies. Schizonepetae Spica (SS), a Chinese medicinal herb, is the dried spike of Schizonepeta tenuifolia Briq. It is available in two forms: raw products and charred products (Schizonepetae Spica Carbonisata, SSC; raw SS processed by stir-frying until carbonization). Raw SS is commonly used to treat TCM symptoms that resemble common cold, fever, respiratory tract infection and allergic dermatitis, while SSC has long been used as a remedy for TCM symptoms that resemble bloody stool and metrorrhagia. AIM OF THE STUDY: We aimed to examine whether stir-fry processing alters the anti-inflammatory, antiviral and hemostatic activities of SS and explore the chemical profile behind the potential changes in medicinal properties caused by stir-fry processing. MATERIALS AND METHODS: We used cell models to examine the anti-inflammatory and antiviral effects of raw SS and SSC. The bleeding time of the tail bleeding model and clotting time of the capillary method in mice were used to compare the hemostasis properties of raw SS and SSC. The chemical profiles of SS and SSC were compared using a method combining gas chromatography-mass spectrometry (GC-MS) and high-performance liquid chromatography/quadrupole-time-of-flight mass spectrometry (HPLC/Q-TOF-MS) analysis. RESULTS: The anti-inflammatory effects of SSC were less potent than those of raw SS. Both raw SS and SSC effectively inhibited viral infection in a dose-dependent manner, with IC50 values of 96.30 and 9.73 µg/mL and selectivity index (SI) values of >1.56 and 7.78, respectively. Interestingly, SSC showed more potent antiviral activities than raw SS. Intragastric administration of raw SS and SSC to mice demonstrated that the hemostatic effects of SSC were more potent than those of raw SS. By comparing the volatile chemical profiles of SSC, we found that twenty-nine constituents disappeared and that fifty-four new constituents were formed while the relative contents of five other components decreased and three other components increased. Additionally, the nonvolatile chemical profiles of raw SS and SSC differed, with thirty-two lower peaks and seven higher peaks in SSC than in SS. CONCLUSION: Our study showed that raw SS and SSC support traditional practice for the clinical applications of these two products except for raw SS used for the treatment of viral infection. It is a fascinating challenge to form SSCs with both traditional hemostatic activities and antiviral properties after stir-fry processing. In addition, the volatile and nonvolatile chemical constituents of raw SS changed dramatically during processing. Further studies are warranted to explore whether the change in chemical constituents is in accordance with the purpose of processing.

Numerical Analysis and Theoretical Verification on the Dynamic Response of Typical Components under Explosion Load.

To analyze the response of float glass under explosion load, the model was developed by the method of FEM simulation combined with stress theory. The numerical simulation results show that the stress caused by the blast wave on the center of the glass panel is continuous, while the stress around the glass is instantaneous. The displacement on both sides of the glass is larger than that at the center and at the four corners of the glass. The effective stress at the center of the glass is 86.08 MPa by numerical simulation. However, the effective stress at the center of the glass is 87.75 MPa by theoretical calculation. The error rate between the simulation result and the theoretical result is only 1.90%. Therefore, it is proved that the theoretical expressions in this paper are reliable for calculating the effective stress of single float glass under explosion load.

The scientific elucidation of daodi medicinal materials.

Daodi medicinal materials (DMMs), with unique characteristics and specific ecological growing environments, are recognized as high-quality medicinal products of Chinese medicinal materials (CMMs). The quality evaluation of CMMs is fundamental for standardization. The concept and application of DMMs have a long history as described in records in ancient books and rooted in practice and experience over generations. DMM is the specific term for pure, superior medicinal herbs with the following characteristics: optimum harvest season (reflecting the appropriate developmental stage of the plant), scrupulous processing, traditional preparation technology, etc. As DMM and high-quality medicinal products are traditionally thought to be closely related, modern scientific studies that confirm the association of these products are described. This article aims to clarify the scientific elucidation of DMMs.

Energy and Vibration Absorption Characteristics of Damping Holes under Explosion Dynamic Loading.

There are usually many buildings near the subway. In tunnel construction, the vibration caused by explosion affects the structure of buildings. The vibration and energy absorption characteristics of damping holes caused by tunnel blasting are investigated under explosion dynamic loading, experimentally and numerically. This paper, taking Yan'an Third Road Station as a case, verified the effectiveness of the damping hole on the vibration reduction of the ground building based on the simulation and experiment. Furthermore, the article examines the effect of distance between the damping hole and charging holes, diameters of the damping hole, and material of the medium on the damping performance. The results show that an increase in diameter improves the energy and vibration absorption performances of the damping hole better than a decrease in distance. In addition, damping performance in gas and liquid medium is better than that in solid medium. The optimum scheme for vibration and energy absorption under extreme conditions of tunnel blasting is to arrange an 80 mm damping hole above the charging holes, and the distance between the damping hole and the center axle wire of the two charging holes is 0.2 m. By using this scheme, the vibration and energy absorption characteristics of damping holes are better than that of other scheme: the damping efficiency is the highest, which is 10.83%, and the pressure-relief efficiency is 10.48%. Furthermore, the medium materials of the damping hole are preferably air or water. Finally, the tunnel excavation contour line is smooth by applying this method. It is concluded that if the damping system is developed with proper design guidelines, then it may reduce the transmission of energy and vibration with effective protection of ground buildings.

[Herbalogical study of Colla Corii Asini(Ejiao)].

Colla Corii Asini(Ejiao)is an important Chinese medicine used in China for thousands of years, and is well known for its famous tonic properties. The herbalogical study was detailed carried out based on the naming, habitat, harvesting, processing, medicinal properties and clinical efficacy. The results showed that the name of Ejiao could be traced back to Shennong's Materia Medica, and various names of Lvpi Jiao, Penfu Jiao and Fuzhi Jiao were recorded in other ancient books. In the many intervening centuries, the main materials of Ejiao had been replaced from cow leather before Tang Dynasty to donkey skin in the middle to late Tang Dynasty. This phenomenon could be probably caused by complicated social factors of various periods and different efficacy of Ejiao made by all kinds of raw materials. Ejiao was merely processed with the simple methods before Tang Dynasty, which subsequently improved avariety of methods to enhance the supplementation action. Most importantly, Ejiao has a wide clinic application along with the development of processing theories and methods, which can be found in various Classics, especially in imperial medical case record in Qing Dynasty.

The Electrical-Triggered High Contrast and Reversible Color-Changing Janus Fabric Based on Double Side Coating.

A sensitive electro-thermochromic Janus fabric driven by voltage is demonstrated via a facial double side coating. The graphene forms a conductive layer that allows Joule heating to supply the thermal resource for the electro-thermochromic behavior of polyester fabric. The thermochromic dye with reversible color-changing property is coated on the opposite side of the graphene layer. The color of electro-thermochromic Janus fabric changes from blue to white with a gradual heating that exceeded 45 °C at the applied voltage of 10 V. The switching rate of color is rapid with the increase of temperature from the room temperature to above 45 °C in 8 s, resulting from the superior resistive heating of the graphene. The electrical conductivity of the electro-thermochromic Janus fabric is not disturbed once undergoing a bending angle range from 30° to 150° and the temperature remains stable after 1000 bending cycles which clearly indicates the excellent flexibility of the fabric. The steady signal in the heating/cooling curve is observed after 500 cycles, pointing out the outstanding durability of the electro-thermochromic Janus fabric under the supplied voltage. It is realizable that the color of electro-thermochromic Janus fabric is triggered accurately by varying the supplied voltage. The simplicity of this design makes it attractive for the application of flexible electro-thermochromic textile, such as active visual camouflage, personal thermal management, and information displays.

Facile synthesis of TiO2/CNC nanocomposites for enhanced Cr(VI) photoreduction: Synergistic roles of cellulose nanocrystals.

In this study, TiO2 nanocrystals were synthesized in the scaffold of cellulose nanocrystal (CNC) using in situ hydrolysis, where the morphology and size of TiO2 was controlled by CNC's functional groups and surface charge. The resulting TiO2/CNC nanocomposites showed a superior photocatalytic activity for Cr(VI) reduction under visible light (λ > 420 nm) due to the combined effects of small TiO2 size and ligand-to-metal charge transfer (LMCT) complex between CNC and TiO2. It was found that the charge-enriched CNC not only acted as a template to direct the crystal growth of TiO2, but also played essential roles on light harvesting and charge transfer thereby promoting the photoreduction of Cr(VI). The demonstrated system represents a unique pathway to develop a lower cost and efficient purification material for remediation of Cr(VI).

Steroid hormone micropollutant removal from water with activated carbon fiber-ultrafiltration composite membranes.

Short activated carbon fibers (ACF) with high surface area were fabricated via carbonization in N2 and activation in CO2 at high temperatures, with cellulose fibers as the raw materials. The obtained ACF were subsequently deposited into the support layer of a polyethersulfone (PES) ultrafiltration membrane by a facile filtration process to obtain the sandwich structured ACF-PES composite membrane. The hormone (17ß-estradiol, E2) adsorption kinetics and isotherm of ACF in static conditions, as well as E2 removal by filtration with the ACF-PES composite membrane were investigated. In static conditions, ACF rapidly and efficiently adsorbs E2 evidenced by a high removal of >97 %. The fitting of second order kinetics and linear (Henry) adsorption isotherm models indicated the availability of easily accessible adsorption sites. Besides, such efficient E2 adsorption was contributed by many interactions between E2 and ACF, namely hydrophobic interactions, hydrogen bonding and π-π stacking. The incorporation of ACF in a PES membrane resulted in a minor loss of filtration flux compared with the control membrane, but significantly improved E2 removal through adsorption pathway. With only 1.0 mg ACF incorporated (loading 2.0 g/m2), the composite membrane could reject 76 % of E2 from a 100 ng/L solution at a flux of 450 L/m2∙h, demonstrating that ACF-PES can overcome the permeability-selectivity trade-off of traditional UF membranes.

Chromatographic fingerprints analysis and determination of seven components in Danmu preparations by HPLC-DAD/QTOF-MS.

BACKGROUND: Danmu preparations (Danmu Capsule and Danmu Syrup), which are made from Nauclea officinalis stem extracts, have good clinical efficacy in acute tonsillitis, acute pharyngitis and upper respiratory tract infection. However, there is currently no reliable and systematic method to control the quality of these two Danmu preparations. METHODS: Using high-performance liquid chromatography (HPLC) coupled with diode array detection (DAD), the fingerprints of the Danmu preparations were established at 250 nm to comprehensively investigate the stability of preparation process. The chemical constituents in the Danmu preparations were separated and identified by HPLC coupled with quadrupole-time-of-flight high-definition mass spectrometry (HPLC-Q-TOF-MS). And seven major components were simultaneously determined at dual wavelengths (250 nm, 326 nm). RESULTS: The results of HPLC fingerprint similarity evaluation showed that the similarity values of 25 batches of Danmu preparations were more than 0.993. Twenty-three compounds, including 10 alkaloids, 6 phenolic acids, 2 iridoids, and 5 unknown compounds, were identified or tentatively characterized according to the retention times and MS/MS fragment patterns of compounds. The developed assay method of seven components was validated with acceptable linearity, precision, repeatability, stability and recovery. The contents of strictosamide belonging to alkaloids as the most abundant constituent in Danmu Capsule and Danmu Syrup were 43,681.20-99,652.49 µg/g and 1567.83-2427.25 µg/mL respectively. The contents of protocatechuic acid which were the highest in measured phenolic acids were 2633.01-7739.78 µg/g in Danmu Capsule and 192.05-448.71 µg/mL in Danmu Syrup, respectively. As an iridoid, the contents of sweroside in Danmu Capsule and Danmu Syrup were 1573.82-2789.81 µg/g and 70.32-182.81 µg/mL, respectively. CONCLUSION: The established qualitative analysis method of fingerprint can be used to attain standardization, uniformity and stability of the preparation process. Meanwhile, the quantitative analysis in this study can be used as an accurate assay method for preparations.

[Recent advances on genuineness evaluation of Lingnan herbal drugs].

Chinese medicine is the traditional treasure of China nation. As the basis of Chinese medicine, traditional Chinese medicine(TCM) plays an important part for the development of Chinese medicine. Genuine medicinal materials with special characteristics of TCM growing in special ecological environment, is recognized as the high quality medicine. Research on genuineness evaluation of TCM is the key to ensure its clinical applications, efficacy and the process of modernization and internationalization for Chinese medicine. Lingnan region of China is situated in the tropical and subtropical zones, where there are rich geothermal and hydrothermal resources. The superior natural and geographic environment of Lingnan has given birth to a variety of native herbal drugs. And treating and preventing diseases with Lingnan herbal drugs has a long story. This study mainly evaluated the genuineness of Lingnan herbal drugs from the aspects of ecological factor, thegenetic information, the history, the culture, the clinical efficacy and the processing, and proposed a new idea to investigate the genuineness of TCM, aiming to provide a scientific basis for genuineness evaluation.

Human Neural Stem Cells with GDNF Site-Specific Integration at AAVS1 by Using AAV Vectors Retained Their Stemness.

The neural stem cells (NSCs) have the ability to self-renew, and to migrate to pathologically altered regions of the central nervous system. Glial cell derived neurotrophic factor (GDNF) could protect dopamine neurons and rescue motor neurons in vivo, which has been proposed as a promising candidate for the treatments of degenerative neurological diseases. In order to combine the advantages of neurotrophic factors and stem cells in clinical therapy, we established the modified hNSCs that has site-specific integration of GDNF gene by using recombinant adeno-associated virus (rAAV) vectors. The hNSCs were co-infected by rAAV2-EGFP-GDNF and rAAV2-SVAV2 which provide integrase to specifically integrate GDNF gene into AAVS1 site. The GDNF-hNSCs maintained their original stem cell characteristics and the ability to differentiate into neurons in vitro. In the animal model, the GDNF-hNSCs were specifically transplanted into CA1 area of hippocampi and could migrate to the dentate gyrus region and differentiate into neuronal cells while maintaining GDNF expression. hNSCs with GDNF gene site-specific integration at AAVS1 by using AAV vectors retained their stemness and effectively expressed GDNF, which indicates the potential of employing transplanted hNPCs for treatment of brain injuries and degenerative neurological diseases.

Characterization and development of EST-derived SSR markers in Sinowilsonia henryi (Hamamelidaceae).

PREMISE OF THE STUDY: Polymorphic microsatellite markers were developed to reveal the genetic diversity of extant populations and the mating system of Sinowilsonia henryi (Hamamelidaceae). METHODS AND RESULTS: In this study, nuclear simple sequence repeat (SSR) markers were developed using the Illumina high-throughput sequencing technique (RNA-Seq). The de novo-assembled transcriptome generated a total of 64,694 unique sequences with an average length of 601 bp. A total of 2941 microsatellite loci were detected. Of the 121 tested loci, 13 loci were polymorphic and eight were monomorphic among 72 individuals representing three natural populations of the species. The number of alleles per locus ranged from one to four, and the observed and expected heterozygosity at population level were 0.00-1.00 and 0.10-0.66, respectively. CONCLUSIONS: The developed expressed sequence tag (EST)-SSRs will be useful for studying genetic diversity of S. henryi as well as assessing the mating system among Sinowilsonia species.