Exosomal miRNA-92a derived from cancer-associated fibroblasts promote invasion and metastasis in breast cancer by regulating G3BP2.

Cancer-associated Fibroblasts (CAFs) exert a tumor-promoting effect in various cancers, including breast cancer. CAFs secrete exosomes containing miRNA and proteins, influencing the tumor microenvironment. In this study, we identified CAF-derived exosomes that transport functional miR-92a from CAFs to tumor cells, thereby intensifying the aggressiveness of breast cancer. CAFs downregulate the expression of G3BP2 in breast cancer cells, and a significant elevation in miR-92a levels in CAF-derived exosomes was observed. Both in vitro and in vivo experiments demonstrate that miR-92a enhances breast cancer cell migration and invasion by directly targeting G3BP2, functioning as a tumor-promoting miRNA. We validated that the RNA-binding proteins SNRPA facilitate the transfer of CAF-derived exosomal miR-92a to breast cancer cells. The reduction of G3BP2 protein by CAF-derived exosomes releases TWIST1 into the nucleus, promoting epithelial-mesenchymal transition (EMT) and further exacerbating breast cancer progression. Moreover, CAF-derived exosomal miR-92a induces tumor invasion and metastasis in mice. Overall, our study reveals that CAF-derived exosomal miR-92a serves as a promoter in the migration and invasion of breast cancer cells by reducing G3BP2 and may represent a potential novel tumor marker for breast cancer.

Maxillary molar distalization treated with clear aligners combined with mini-implants and angel button using different traction force: a finite element study.

OBJECTIVES: To evaluate the biomechanical system of molar distalization with clear aligner therapy (CAT) combined with angel button using interradicular mini-implants (IRMIs) with varying elastic forces. MATERIALS AND METHODS: FE models including maxilla, complete maxillary dentition, periodontal ligaments (PDL), composite attachments, mini-implants (MI), and dedicated orthodontic aligner, were constructed. Three groups were created in accordance with the sagittal position of MI. Elastic forces (0 N,1 N,1.5 N,2 N) were applied. RESULTS: CAT without elastics caused labial tipping and intrusion of the anterior teeth. Initial labial tipping and the von Mises stress of the maxillary anterior teeth decreased as the elastic forces increased.

[Prokaryotic expression and biological activities of the hemolysin BL subunit of a pathogenic Bacillus cereus of cattle origin].

Bacillus cereus belongs to Gram-positive bacteria, which is widely distributed in nature and shows certain pathogenicity. Different B. cereus strains carry different subsets of virulence factors, which directly determine the difference in their pathogenicity. It is therefore important to study the distribution of virulence factors and the biological activity of specific toxins for precise prevention and control of B. cereus infection. In this study, the hemolysin BL triayl was expressed, purified, and characterized. The results showed that the bovine pathogenic B. cereus hemolysin BL could be expressed and purified in the prokaryotic expression system, and the bovine pathogenic B. cereus hemolysin BL showed hemolysis, cytotoxicity, good immunogenicity and certain immune protection in mice. In this study, the recombinant expression of hemolysin BL triayl was achieved, and the biological activity of hemolysin BL of bovine pathogenic ceroid spore was investigated. This study may facilitate further investigating the pathogenic mechanism of B. cereus hemolysin BL and developing a detection method for bovine pathogenic B. cereus disease.

Exosome-based engineering strategies for the diagnosis and treatment of oral and maxillofacial diseases.

Oral and maxillofacial diseases are one of the most prevalent diseases in the world, which not only seriously affect the health of patients' oral and maxillofacial tissues, but also bring serious economic and psychological burdens to patients. Therefore, oral and maxillofacial diseases require effective treatment. Traditional treatments have limited effects. In recent years, nature exosomes have attracted increasing attention due to their ability to diagnose and treat diseases. However, the application of nature exosomes is limited due to low yield, high impurities, lack of targeting, and high cost. Engineered exosomes can be endowed with better comprehensive therapeutic properties by modifying exosomes of parent cells or directly modifying exosomes, and biomaterial loading exosomes. Compared with natural exosomes, these engineered exosomes can achieve more effective diagnosis and treatment of oral and maxillary system diseases, and provide reference and guidance for clinical application. This paper reviews the engineering modification methods of exosomes and the application of engineered exosomes in oral and maxillofacial diseases and looks forward to future research directions.

Synthesis of multicolor luminescent carbon dots based on carboxymethyl chitosan for cell imaging and wound healing application: In vitro and in vivo studies.

The construction of biomaterials that can facilitate wound healing is significantly challenging in the medical field, and bacterial infections increase this complexity. In this study, we selected the biomacromolecule carboxymethyl chitosan as a carbon source and citric acid as an auxiliary carbon source. We prepared carbon quantum dots with multicolor luminescence properties and higher quantum yields (QYs) using a facile one-pot hydrothermal method. We characterized them to select carbon dots (CDs) suitable for cell growth. Subsequently, their biocompatibility with L929 cells, antibacterial properties against Staphylococcus aureus, and efficiency in promoting wound healing in vivo were investigated. Our experimental results showed that CDs at an appropriate concentration had excellent bioimaging ability, were suitable for cell growth, and accelerated the healing of infected wounds. We believe these bioactive CDs have great potential in promoting wound healing.

SIRT6 Knockdown in Buffalo Fetal Fibroblasts Exacerbates Premature Senescence Caused by DNA and Telomere Damage.

As a gene with antiaging functions, sirtuin6 (SIRT6) belonging to the sirtuin family plays a vital role in DNA repair, telomerase function, and cellular senescence, as well as maintains epigenomic stability and promotes longevity. However, its role in cell senescence in large animals, such as buffaloes, remains unknown. Fibroblasts are commonly used for somatic reprogramming, and their physiological characteristics affect the efficiency of this process. We aimed to elucidate the role of SIRT6 in cellular senescence and proliferation and analyze its effect on the biological function of buffalo fibroblasts to help improve the efficiency of buffalo somatic cell reprogramming. The expression of SIRT6 and related DNA damage was measured in buffalo fibroblasts obtained at different developmental stages (in the fetus and at 3 and 10 years of age), and the effect of SIRT6 knockdown on the senescence of buffalo fetal fibroblast was investigated. An inverse relationship was observed between SIRT6 expression and senescence in buffalo fibroblasts obtained from animals of various ages. This was accompanied by decreased cell growth, viability, and increased DNA damage. Short hairpin RNA-mediated SIRT6 knockdown accelerated the senescence of buffalo fetal fibroblasts. It blocked the cell cycle during in vitro cell culture, which further enhanced DNA damage, particularly with respect to the telomeres. Collectively, our findings suggest that SIRT6 expression was closely associated with buffalo senescence in fibroblasts. These findings serve as a foundation to better understand the cellular functions of SIRT6 and also aid in selecting donor cells for buffalo somatic cell reprogramming.

Biosafety evaluation of BaSi2O2N2:Eu2+/PDMS composite elastomers.

In recent years, mechanoluminescent (ML) materials have shown great potential in stress sensing, mechanical energy collection and conversion, so they have attracted wide attention in the field of stomatology. In the early stage of this study, BaSi2O2N2:Eu2+ ML phosphors were synthesized by two-step high temperature solid state method, and then mixed with Polydimethylsiloxane (PDMS) in different proportions to obtain BaSi2O2N2:Eu2+/PDMS ML composites with different mass fractions (10%,20%,30%,40%,50%). Then its biosafety was evaluated by Cell Counting Kit-8 (CCK-8), Calcein-AM/PI fluorescence staining, hemolysis, oral mucosal irritation, acute and subacute systemic toxicity tests. The experimental results show that the biosafety of BaSi2O2N2:Eu2+/PDMS ML composite elastomers with different mass fraction is in line with the existing standards, and other related properties can be further studied.

Application of Flow Cytometry in the Diagnosis of Bovine Epidemic Disease.

As science and technology continue to advance, the use of flow cytometry is becoming more widespread. It can provide important information about cells in the body by detecting and analysing them, thereby providing a reliable basis for disease diagnosis. In the diagnosis of bovine epidemic diseases, flow cytometry can be used to detect bovine viral diarrhoea, bovine leukaemia, bovine brucellosis, bovine tuberculosis, and other diseases. This paper describes the structure of a flow cytometer (liquid flow system, optical detection system, data storage and analysis system) and its working principles for rapid quantitative analysis and sorting of single cells or biological particles. Additionally, the research progress of flow cytometry in the diagnosis of bovine epidemic diseases was reviewed in order to provide a reference for future research and application of flow cytometry in the diagnosis of bovine epidemic diseases.

Finite element analysis of tooth movement under different retraction force and intrusive force in double-archwire lingual orthodontics system.

The objectives of this study were to evaluate the tooth movement tendency during space closure in maxillary anterior teeth by various combinations of retraction force and intrusive force in a double-archwire lingual orthodontic system. Mini-implant-double slot lingual orthodontics system models of the bilateral maxillary first premolar extraction cases were constructed. Three-dimensional finite element models of the maxilla were constructed with definite position mini-implants (8 mm) and power arms (6 mm). Different retraction forces(50gf、100gf、150gf)were applied with the help of a nickel-titanium closed coil spring on the plate side. Intrusive forces(0gf、50gf、100gf)were applied with the help of the mini-implant between the two central incisors, and the initial displacements of the maxillary anterior teeth were analyzed. Variable amounts of displacements like controlled tipping, uncontrolled tipping, lingual crown tipping, labial root tipping, extrusion and distal crown tipping were observed in all the models, and these tendencies increased as the magnitude of retraction force increased, and these tendencies decreased as the magnitude of intrusive force increased. When the intrusive force was greater than or equal to the retraction force, the maxillary central incisors showed the trend of lingual crown tipping and labial root tipping, resulting in uncontrolled tipping movement. In terms of horizontal changes, the increasing width of bilateral anterior teeth was observed, with canines showing the least increasing trend. Various combinations of retraction force and intrusive force in a double-archwire lingual orthodontic system provide a new choice for torque control of the anterior teeth. Although anterior mini-implants and elastics can achieve incisor intrusion and lingual root torque, they cannot achieve the expected torque without additional torque control methods.

Inhibitory Effect and Mechanism of Ursolic Acid on Cisplatin-Induced Resistance and Stemness in Human Lung Cancer A549 Cells.

The survival rate of lung cancer patients remains low largely due to chemotherapy resistance during treatment, and cancer stem cells (CSCs) may hold the key to targeting this resistance. Cisplatin is a chemotherapy drug commonly used in cancer treatment, yet the mechanisms of intrinsic cisplatin resistance have not yet been determined because lung CSCs are hard to identify. In this paper, we proposed a mechanism relating to the function of ursolic acid (UA), a new drug, in reversing the cisplatin resistance of lung cancer cells regulated by CSCs. Human lung cancer cell line A549 was selected as the model cell and treated to become a cisplatin-resistant lung cancer cell line (A549-CisR), which was less sensitive to cisplatin and showed an enhanced capability of tumor sphere formation. Furthermore, in the A549-CisR cell line expression, levels of pluripotent stem cell transcription factors Oct-4, Sox-2, and c-Myc were increased, and activation of the Jak2/Stat3 signaling pathway was promoted. When UA was applied to the cisplatin-resistant cells, levels of the pluripotent stem cell transcription factors were restrained by the inhibition of the Jak2/Stat3 signaling pathway, which reduced the enrichment of tumor stem cells, and in turn, reversed cisplatin resistance in lung cancer cells. Hence, as a potential antitumor drug, UA may be able to inhibit the enrichment of the lung CSC population by inhibiting the activation of the Jak2-Stat3 pathway and preventing the resistance of lung cancer cells to cisplatin.

The influence of time structure on prediction motion in visual and auditory modalities.

Usually people can estimate the correct position of a moving object even when it temporarily moves behind an occlusion. Studies have been performed on this type of occluded motion with prediction motion (PM) tasks in the laboratory. Previous publications have emphasized that people could use mental imagery or apply an oculomotor system to estimate the arrival of a moving stimulus at the target place. Nevertheless, these two ways cannot account for the performance difference under a different set of conditions. Our study tested the role of time structure in a time-to-collision (TTC) task using visual and auditory modalities. In the visual condition, the moving red bar travelled from left to right and was invisible during the entire course but flashed at the initial and the occluded points. The auditory condition and visual condition were alike, except that the flashes in the visual condition were changed to clicks at the initial and the occluded points. The results illustrated that participants' performance was better in the equal time structure condition. The comparison between the two sense modalities demonstrated a similar tendency, which suggested there could be common cognitive processes between visual and auditory modalities when participants took advantage of temporal cues to judge TTC.

Morphology changes of maxillary molar distalization by clear aligner therapy / Cambios de la morfología en la distalización de molares maxilares mediante terapia con alineadores transparentes

SUMMARY: To evaluate the skeletal, dento-alveolar and soft tissue morphology changes after maxillary molar distalization by clear aligner therapy and identify the significant efficacy of molar distalization,18 patients in conformity with the inclusion criteria were selected. Pre- and post-treatment Cone Beam Computed Tomography (CBCT) were examined to measure the angular and linear parameters. All subjects were completed non-extraction clear aligner treatment by distalizing molars. A paired-t test and independent-samples t-test were performed to observe the difference between before and after treatment and the difference between the first molar and second molar respectively. P-values <0.05 were considered statistically significant. Predicted movement rate was calculated by the formula: (actual movement(mm)/planned movement(mm)) x100%. Most variables of pre- and post-treatment showed no statistical difference(P<0.05), excepting SNA angle (P<0.05) and Upper lip/E-line linear (P<0.01) due to incisor retraction. The first and second molar revealed a translation movement without significant tipping and vertical movement. Clear aligners provided a high predictability (83.44 %) of distalization the maxillary first molar, and 85.14 % of the maxillary second molar. Clear aligners can effectively achieve distal displacement of molars.

RESUMEN: Se seleccionaron 18 pacientes, de acuerdo con los criterios de inclusión, para evaluar los cambios en la morfología esquelética, dentoalveolar y de los tejidos blandos después de la distalización de los molares maxilares, mediante la terapia con alineadores transparentes e así identificar la significativa eficacia de la distalización de los molares. Se examinó a través de tomografía computarizada de haz cónico (CBCT) antes y después del tratamiento para medir los parámetros angulares y lineales. Todos los sujetos completaron el tratamiento con alineadores transparentes sin extracción mediante la distalización de los molares. Se realizó una prueba t pareada y una prueba t de muestras independientes para observar la diferencia entre antes y después del tratamiento y la diferencia entre el primer molar y el segundo molar, respectivamente. Los valores de p<0,05 se consideraron estadísticamente significativos. La tasa de movimiento prevista se calculó mediante la fórmula: (movimiento real (mm)/movimiento planificado (mm)) x 100 %. La mayoría de las variables de pre y postratamiento no mostraron diferencia estadística (P<0,05), excepto el ángulo SNA (P<0,05) y el labio superior/línea E lineal (P<0,01) debido a la retracción del incisivo. El primer y segundo molar revelaron un movimiento de traslación sin inclinación significativa y movimiento vertical. Los alineadores transparentes proporcionaron una alta previsibilidad (83,44 %) de la distalización del primer molar superior y del 85,14 % del segundo molar superior. Los alineadores transparentes pueden lograr efectivamente el desplazamiento distal de los molares.

Short-term cost-effectiveness of oral semaglutide for the treatment of type 2 diabetes mellitus in the United States.

BACKGROUND: Oral semaglutide is the first orally administered glucagon-like peptide-1 receptor agonist (GLP-1RA) approved by the FDA. Clinical trials found that oral semaglutide 14 mg had a greater reduction in hemoglobin A1c (A1c) compared with empagliflozin 25 mg and sitagliptin 100 mg and was noninferior to liraglutide 1.8 mg. However, US cost-effectiveness data for oral semaglutide are limited and do not consider the costs of adverse events. OBJECTIVE: To assess the short-term cost-effectiveness of oral semaglutide compared with empagliflozin, sitagliptin, and liraglutide in patients with type 2 diabetes. METHODS: A decision analysis over a 52-week time horizon was used to evaluate the incremental cost-effectiveness of oral semaglutide vs empagliflozin, sitagliptin, and liraglutide from a US health care payer's perspective. Data on efficacy, adverse events, and discontinuation were derived from 52-week data from phase 3, head-to-head clinical trials (PIONEER 2, 3, and 4). Costs included drug and administration cost and treatment of gastrointestinal adverse events. Incremental cost-effectiveness ratios (ICERs) were calculated as the difference in cost over the difference in A1c reduction between oral semaglutide and comparators. RESULTS: In the base-case analysis, 52-week treatment costs with oral semaglutide were $2,660 and $3,104 higher and $2,337 less than empagliflozin, sitagliptin, and liraglutide, respectively. Incremental (greater) A1c reductions were seen with oral semaglutide at 0.40%, 0.50%, and 0.30% vs empagliflozin, sitagliptin, and liraglutide, respectively. ICERs per 1% reduction in A1c for oral semaglutide were $6,650 and $6,207 vs empagliflozin and sitagliptin, respectively. Oral semaglutide was dominant vs liraglutide (ICER of -$7,790). CONCLUSIONS: Oral semaglutide was dominant relative to liraglutide, offering a cost-saving GLP-1RA oral alternative. While there is not a recognized willingness-to-pay threshold for a 1% reduction in A1c, oral semaglutide may be cost-effective relative to empagliflozin and sitagliptin if a decision maker's willingness-to-pay threshold exceeds $6,650 and $6,207, respectively. DISCLOSURES: No outside funding supported this study. The authors have no conflicts of interest to declare.

A Clinical Analysis of the Treatment of Chronic Coronary Artery Occlusion With Antegrade Dissection Reentry.

Objective: This study aimed to investigate the efficacy and safety of antegrade dissection re-entry (ADR) technique in the percutaneous coronary intervention (PCI) to open chronic total occlusion (CTO) lesions. Methods: The baseline, angiographic results, PCI success rate, and major adverse cardiac events (MACE) during the 12 months of follow-up were compared between 48 patients who did not use ADR in the treatment of CTO lesions (control group) and 50 patients who used ADR (treatment group). Results: The control group comprised 48 patients who had 52 CTO lesions, and the treatment group comprised 50 patients who had 58 CTO lesions. The success rate of PCI in the treatment group (89.7 vs. 71.2%, P = 0.047) was significantly higher than in the control group, where six patients had in-stent restenosis (ISR, ISR-CTO) that were all recanalized. The mean PCI time (71 ± 25 min vs. 95 ± 33 min, P = 0.041), X-ray exposure time (42 ± 17 min vs. 71 ± 22 min, P = 0.032), contrast agent dosage (98 ± 26 ml vs. 178 ± 63 ml, P = 0.029), MACE incidence during the 12 months of follow-up (22.0 vs. 41.7%, P = 0.046) and recurrent myocardial infarction incidence (10.0 vs. 27.1%, P = 0.047) were significantly lower in the treatment group than in the control group. The differences were all statistically significant. Conclusion: It is safe and effective to use the ADR technique in PCI for coronary artery CTO lesions. The technique shortens the operation time, reduces the radiation dose of doctors and patients and the use dose of contrast agents, and improves patients' prognoses.

[SP13786 Inhibits the Migration and Invasion of Lung Adenocarcinoma Cell A549 
by Supressing Stat3-EMT via CAFs Exosomes].

BACKGROUND: Fibroblast activation protein (FAP) is one of the surface markers of cancer-associated fibroblasts (CAFs) and is closely related to the malignant characterization of CAFs. SP13786 is a specific micromolecule inhibitor of FAP and this study is to investigate the effects and mechanism of SP13786 on the migration and invasion of A549 cells through regulating exosomes of CAFs. METHODS: CAFs and paracancerous fibroblasts (PTFs) were isolated and subcultured from freshly resected lung adenocarcinoma tissues and paracancerous normal tissues separately. MTT assay was used to detect the proliferation of CAFs incubated by different concentrations of SP13786; PTFs-exo, CAFs-exo and CAFs+SP13786-exo were extracted by polymer precipitation method. The A549 cells were divided into Ctrl group, PTFs group, CAFs group and SP13786 group and each group was incubated with DMEM, PTFs-exo, CAFs-exo and CAFs+SP13786-exo separately. Laser confocal microscope was used to observe the endocytoses of exosomes by A549 cells. The expression of alpha-smooth muscle actin (α-SMA) and FAP in PTFs and CAFs and the expression of E-cadherin, N-cadherin, Slug, Stat3 and P-Stat3 in A549 cells were detected by immunofluorescence, immunohistochemistry and Western blot. The migration and invasion ability of A549 cells were detected by cell scratch and transwell methods. RESULTS: α-SMA and FAP were expressed much higher in CAFs than that in PTFs which indicate that CAFs and PTFs were successfully obtained from lung adenocarcinoma and paracancerous tissues (P<0.05). MTT showed that the 50% inhibitory concentration (IC50) of SP13786 for CAFs was about 3.3 nmol/L. In addition, SP13786 can significantly decrease the expression of α-SMA and FAP in CAFs which means that targeted inhibition of FAP could reduce the malignant characteristics of CAFs (P<0.05). Laser confocal microscope found that exosomes from CAFs could be taken up by A549 cells and scratch and transwell tests showed that the endocytosed CAFs-exo could promote the migration and invasion of A549 cells (P<0.001), while FAP inhibitor SP13786 could inhibit the effects of CAFs-exo on A549 cells (P<0.05). Furthermore, Immunofluorescence and Western blot showed that CAFs-exo could promote EMT by decreasing E-cadherin expression and increasing N-cadherin, Slug expression in A549 cells while FAP inhibitor SP13786 could significantly supress CAFs-exo-induced epithelial-mesenchymal transition (EMT) of A549 cells (P<0.05). Moreover, the expression of P-Stat3 was obviously increased in A549 cells of CAFs group and significantly down-regulated in SP13786 group (P<0.05) whereas there was no significant difference in total Stat3 between CAFs and SP13786 groups (P>0.05). Finally, WP1066 (a specific inhibitor of Stat3) was used to comfirm whether SP13786 could influence EMT of A549 cells by inhibiting Stat3 phosphorylation via CAFs-Exo. The results showed that when the phosphorylation of Stat3 in CAFs group was inhibited by WP1066, SP13786 could not influence the P-Stat3 expression and EMT of A549 cells anymore (P>0.05). CONCLUSIONS: As a specific micromolecule inhibitor of FAP, SP13786 indirectly inhibits the migration and invasion of A549 cells by affecting exosomes of CAFs. The possible mechanism is to inhibit the phosphorylation of Stat3 and thus affect the EMT of A549 cells.

Hypoxia enhances buffalo adipose-derived mesenchymal stem cells proliferation, stemness, and reprogramming into induced pluripotent stem cells.

Adipose tissue-derived mesenchymal stem cells (ASCs) from livestock are valuable resources for animal reproduction and veterinary therapeutics. Previous studies have shown that hypoxic conditions were beneficial in maintaining the physiological activities of ASCs. However, the effects of hypoxia on buffalo ASCs (bASCs) remain unclear. In this study, the effects of hypoxia on proliferation, stemness, and reprogramming into induced pluripotent stem cells (iPSCs) of bASCs were examined. The results showed that the hypoxic culture conditions (5% oxygen) enhanced the proliferation and colony formation of bASCs. The expression levels of proliferation-related genes, and secretion of basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF) were significantly enhanced in hypoxia. Hypoxic culture conditions activated hypoxia-inducible factor-1α (HIF-1α), thereby contributing to the secretion of bFGF and VEGF, which in turn enhanced the expression of HIF-1α and promoted the proliferation of bASCs. Furthermore, in hypoxic culture conditions, bASCs exhibited the main characteristics of mesenchymal stem cells, and the expression levels of the pluripotent markers OCT4, NANOG, C-MYC, and the differentiation capacity of bASCs were significantly enhanced. Finally, bASCs were more efficiently and easily reprogrammed into iPSCs in hypoxic culture conditions and these iPSCs exhibited some characteristics of naïve pluripotent stem cells. These findings provide the theoretical guidance for elucidating the detailed mechanism of hypoxia on physiological activities of bASCs including proliferation, stemness maintenance, and reprogramming.

Isolation and characterization of buffalo (bubalus bubalis) amniotic mesenchymal stem cells derived from amnion from the first trimester pregnancy.

Amniotic mesenchymal stem cells (AMSCs) from livestock are valuable resources for animal reproduction and veterinary therapeutic. The purpose of this study is to explore a suitable way to isolate and culture the buffalo AMSCs (bAMSCs), and to identify their biological characteristics. Digestion with a combination of trypsin-EDTA and collagenase type I could obtain pure bAMSCs more effectively than trypsin-EDTA or collagenase type I alone. bAMSCs could proliferate steadily in vitro culture and exhibited fibroblastic-like morphology in vortex-shaped colony. bAMSCs were positive for MSC-specific markers CD44, CD90, CD105, CD73, ß-integrin (CD29) and CD166, and pluripotent markers OCT4, SOX2, NANOG, REX-1, SSEA-1, SSEA-4 and TRA-1-81, but negative for hematopoietic markers CD34, CD45 and epithelial cells specific marker Cytokeratin 18. In addition, bAMSCs were capable of differentiating into adipogenic, osteogenic, chondrogenic and neural lineages, with expression of FABP4, Ost, ACAN, COL2A1, Nestin and ß III-tubulin. Glycogen synthase kinase 3 inhibitor: kenpaullone promoted bAMSCs to differentiate into neural lineage. This study provides an effective protocol to obtain and characterize bAMSCs, which have proven useful as a cell resource for buffalo cell reprogramming studies and transgenic animal production.

Molecular Cloning, Identification, and Expression Patterns of Myostatin Gene in Water Buffalo (Bubalus Bubalis).

Myostatin (MSTN), also named growth differentiation factor 8 (GDF8), is a transforming growth factor-ß (TGF-ß) family member with a key role in the negative regulation of skeletal muscle growth. However, its role in ovarian folliculogenesis remains unclear. To provide us with a basis for understanding this role, we cloned MSTN and examined its expression patterns in water buffalo (Bubalus bubalis). The complete ORF of the water buffalo MSTN gene is 1,128 nucleotides, which encode a 375 amino acid protein and sharing 99% identity at the deducted amino acid level with that of Bos taurus. Protein sequence analysis showed that MSTN is a weakly acerbic extracellular protein, consisting of signal peptides at 18-19 sites, a TGF-ß propeptide, and a TGF-ß domain. RT-PCR analyses demonstrated that water buffalo MSTN was expressed in multiple tissues but not limited to muscle. Immunohistochemistry staining confirmed the presence of MSTN in oocytes and granulosal cells. To our knowledge, this is the first study to confirm the expression of MSTN in the water buffalo ovary, suggesting an additional role of MSTN in water buffalo folliculogenesis, along with its role in skeletal muscle growth regulation. Further study of the regulatory mechanism of MSTN in water buffalo reproduction is warranted. ABBREVIATIONS: MSTN, myostatin; ORF, open reading frame.

Diphenyl ethers from Aspergillus sp. and their anti-Aß42 aggregation activities.

Two new compounds with the character of diphenyl ether structure, oxisterigmatocystin D (1) and 9-acetyldiorcinol B (6), were isolated from the endolichenic fungal strain Aspergillus sp. (No. 16-20-8-1), along with six known compounds, oxisterigmatocystin A (2), oxisterigmatocystin C (3), sterigmatocystin (4), diorcinol B (5), violaceol-I (7), and violaceol-II (8). The structures of the new compounds were determined by extensive NMR spectroscopic data, and the absolute configuration of 1 was established by single-crystal X-ray diffraction analysis. Moreover, the Aß42 aggregation inhibitory activities of 5-8 were evaluated by the standard thioflavin T (ThT) fluorescence assay using epigallocatechin gallate (EGCG) as the positive control. Compounds 7 and 8 displayed significant anti-Aß42 aggregation activity with IC50 values of 5.1 and 2.3µM, respectively. Preliminary structure-activity relationship of these diphenyl ethers as anti-Aß42 aggregation inhibitors was proposed.

Nodulisporisteriods A and B, the first 3,4-seco-4-methyl-progesteroids from Nodulisporium sp.

Two new 4-methyl-progesteroids, nodulisporisteriod A (1) and nodulisporisteriod B (2), were isolated from the extract of an endolichenic fungal strain Nodulisporium sp. (No. 65-17-2-1), along with two related metabolites, demethoxyviridin (3) and inoterpene B (4). Their structures were determined by detailed spectroscopic analyses, X-ray crystallographic analysis and comparison of the NMR data with those of the closely related compounds previously reported. Nodulisporisteriod A (1) and nodulisporisteriod B (2) possess new carbon skeletons, which are the first cases of fission at C-3,4 in 4-methyl-progesteroids. A hypothetical biosynthetic pathway for 1 and 2 was proposed. Moreover, the Aß42 aggregation inhibitory activities of 1-4 were evaluated using standard thioflavin T (ThT) fluorescence assay with epigallocatechin gallate (EGCG) as positive control. Demethoxyviridin (3) displayed anti-Aß42 aggregation activity with IC50 value of 13.4µM.