Vineyard soil heterogeneity and harvest date affect volatolomics and sensory attributes of Cabernet Sauvignon wines on a meso-terroir scale.

To produce premium wines in a specific region is the goal of local oenologists. This study aimed to investigate the influence of soil properties and harvest date on the volatolomics of wine to provide a better insight into single-vineyard wines. Six Cabernet Sauvignon vineyards were selected in a semi-arid region to produce their wines at three harvest ripeness levels ranging from 23°Brix-28°Brix in three seasons (2019-2021). Results showed that among all six vineyards, the vineyard with the highest soil pH produced wines with lower C6 alcohols and herbaceous aroma. Moderate nutrition in soils was beneficial for the accumulation of ß-damascenone and enhanced fruity and floral aroma in wines while over-fertile soil produced wines with the lowest sensory score. As the harvest ripeness elevated, the wine's fruity and floral aroma intensity decreased. Through advanced network analysis, the key volatiles such as ß-damascenone, ethy1 lactate, and isoamyl octanoate, and their interaction in affecting wine sensory scores were evaluated. Our study provided a concept for producing premium single-vineyard wines.

The key role of vineyard parcel in shaping flavonoid profiles and color characteristics of Cabernet Sauvignon wines combined with the influence of harvest ripeness, vintage and bottle aging.

Recently, revealing the terroir influence on wine chemical features has drawn increasing interest. This study aimed to explain how wine flavonoid signatures were altered by vineyard parcel, harvest ripeness, vintage and bottle aging. Six commercial Cabernet Sauvignon vineyards were selected in the Manas region to produce wines at three harvest ripeness in three seasons (2019-2021) and aged for three years. The six vineyards had little difference in mesoclimate conditions while varying greatly in soil composition. Results showed high vineyard pH (> 8.5) could accelerate grape ripening rate and increase wine flavonol concentration. Vineyards with moderate nutrition produced wines with abundant anthocyanin derivatives and maintained color characteristics during aging. The role of detailed anthocyanin derivatives in regulating wine color was clarified. As the harvest ripeness elevated, wine's flavonoid profiles were altered and gained a higher red color intensity. This work provides chemical mechanisms underlying single-vineyard wines and a theoretical basis for targeted wine production.

Manipulating the severe shoot topping delays the harvest date and modifies the flavor composition of Cabernet Sauvignon wines in a semi-arid climate.

This study aimed to mitigate the adverse effects of global warming by applying severe shoot topping (SST) to grapevines in a semi-arid climate. A three-year study (2018-2020) was performed to investigate the impact of SST on wine flavor composition. Results showed that SST effectively delayed the grape harvest date, which was more evident in the dry and warm vintage (7-11 d). SST significantly increased the concentration of myricetin-based flavonols in wines which were 18% higher than in untreated wines. Through orthogonal partial least squares discriminant analysis (OPLS-DA), SST wines were characterized by more abundant phenolic compounds and higher sensory scores. The carry-over effect of applying SST in consecutive years in the same vines could be reflected in wine color. The correlations among wine metabolites, color and aroma parameters, and sensory parameters were evaluated through multiple analyses. This study provided an idea for delaying grape ripening in a semi-arid climate.

Volatomics of 'Cabernet Sauvignon' grapes and wines under the fan training system revealed the nexus of microclimate and volatile compounds.

The cluster-zone of the fan training system with multiple trunks (F-MT) distribute >1.5 m vertically and 1.0 m horizontally, which leads to microclimate heterogeneity around clusters. In the current study, clusters were divided into eight spatial positions according to the cluster growing height and light conditions, and the aroma profiles of grapes and wines were evaluated by HS-SPME-GC-MS. Results showed that the microclimate varied in different spatial positions. Light exposure promotes the accumulation of terpenes and C13-norisoprenoids, while inhibiting C6/C9 compounds in grape berries. Zone 2 wine presented the highest global aroma concentration. Floral and fruity were the main aromas in F-MT wines, but the herbaceous was more prominent in lower-position wines. C6/C9 compounds in grapes negatively correlated with terpenes and C13-norisoprenoids in wines. In conclusion, the aroma profiles of grapes and wines varied from different cluster positions, and graded harvesting will be helpful to produce quality wines.

Reducing the source/sink ratio of grapevine to face global warming in a semi-arid climate: Effects on volatile composition of Cabernet Sauvignon grapes and wines.

The heterogeneity of the vineyard environment caused high variability in grape metabolites and flavor profiles, and the phenomenon was more prominent in recent years of climate change. Herein, distal leaf removal was applied in semi-arid Xinjiang to adjust the source to sink ratio of grapevines for three consecutive years (2018-2020). The grape-derived volatiles showed high correlations with specific climate factors such as temperature changes in the growth period. Results showed that distal leaf removal increased the solar radiation reaching the clusters in the first few days after applying LR treatments while not affecting the temperature. The improvement in fruity and floral aroma intensity by distal leaf removal was founded not only in grape metabolites but also in wines. Moderate cluster exposure brought by distal leaf removal was beneficial for the accumulation of isoprenoids, which therefore increased the fruity and floral intensity of wines. The carry-over effect did not show in consecutively defoliated vines among vintages regarding the wine aroma profile.

Cluster spatial positions varied the phenolics profiles of 'Cabernet Sauvignon' grapes and wines under a fan training system with multiple trunks.

The fan training system with multiple trunks (F-MT) is widely used in the northern China winegrape region with high yield and convenience for burying soil to prevent winter cold. However, the wide distribution of clusters under F-MT usually leads to variations in berry compositions. In two consecutive years, clusters from different spatial positions were collected to determine the phenolic composition in grapes and resultant wines by HPLC-QqQ-MS/MS. Results showed that light exposure promotes the accumulation of flavonols and 3'-hydroxylated anthocyanins in berries. Wines made from upper clusters had higher alcohol degree, lightness, monomeric anthocyanins and flavonols, whereas wines made from lower clusters had relatively higher total acidity, red color component, polymeric anthocyanin and total flavanols. Most of the wine parameters were also significantly affected by vintage. In conclusion, the quality of grapes and wines varied from different cluster positions, graded harvesting helps to maximize the characteristics of grape berries.

Morphological and Molecular Characterization of Paragonimus skrjabini Complex from Yunnan, China: A Brief Report.

PURPOSE: To perform environmental sampling and molecular identification of Paragonimus in endemic regions, which may help in minimizing transmission among humans. METHODS: Mountain crabs from the genus Potamiscus were collected and the encysted metacercariae were extracted and subjected to morphological identification, followed by animal inoculation in Sprague-Dawley (SD) rats. After 112 days of infection, animals were killed and adult worms were extracted from lungs and muscles. The morphology of adult worms was characterized by microscopy and molecular identification was done by polymerase chain reaction, followed by sequencing of cox1 and ITS2 genes. Phylogenetic analysis was done by maximum parsimony method. RESULTS: A total of 447 crabs were captured from the streams of Tongchang Town, Jinping County, Yunnan Province, China. The infection rate was found to be 41% (186 out of 447 crabs). The metacercariae of Paragonimus skrjabini was identified by the characteristics round or spherical encysted form measuring 410 to 460 × 400 to 460 µm. After animal infection in SD rats, adults were presumptively confirmed to be P. skrjabini, which was also confirmed by gene amplification and sequence analysis of cox1 and ITS2 regions. Paragonimus skrjabini clustered with previously reported P. skrjabini from Yunnan and Vietnam. The confidence values of their branches were > 95%. Phylogenetic analysis of the ITS2 region revealed two distinct clusters with distinct geographical grouping. Phylogenetic analysis with the combined data sets reiterated the geographical grouping with P. skrjabini from Yunnan clustering with strains from Vietnam. CONCLUSION: Metacercariae of P. skrjabini was discovered in freshwater crabs in Yunnan province, China, and the strains were phylogenetically related to P. skrjabini from Vietnam.

Distal leaf removal made balanced source-sink vines, delayed ripening, and increased flavonol composition in Cabernet Sauvignon grapes and wines in the semi-arid Xinjiang.

Leaf removal applied in the upper canopy of modified vertical shooting positioning trellis system has been proposed as an effective strategy to mitigate the adverse effects of global warming on grape and wine quality. In this study, we removed the upper leaves of Cabernet Sauvignon canopy in a semi-arid climate for three consecutive years (2018-2020). About one-third of the whole canopy leaves were removed at the beginning of véraison (LR1) and post-véraison (LR2). All leaf removal treatments included two schemes: (i) leaf removal in the same vines in all vintages to investigate the carry-over effects (1-LR1 and 1-LR2); (ii) leaf removal in different vines in each vintage as repeated experiments among vintages (2-LR1 and 2-LR2). Results showed that leaf removal treatments significantly decreased total soluble solids accumulation in grapes without affecting titratable acidity and pH. LR1 treatments could delay ripening to 6.6 days on average, which was 2.6 days longer than LR2 treatments. LR treatments did not affect the yield but decreased soluble sugar content in canes. Leaves net assimilation rate showed no compensation for the loss of leaves. For phenolic composition, LR treatments increased flavonol concentration in both wines and grapes while had inconsistent effects on anthocyanins and flavanols over three seasons. Principal component analysis (PCA) showed that different LR treatment stages (LR1s vs LR2s) and whether LR in the same vines over consecutive years (1-LRs vs 2-LRs) had limited effects on phenolic profiles. In conclusion, LR in consecutive years at the upper canopy of grapevines was a practical strategy to face global warming in Xinjiang.

Microcliamte changes caused by black inter-row mulch decrease flavonoids concentrations in grapes and wines under semi-arid climate.

In this study, black geotextile inter-row mulch, a weed control practice, was applied under a semi-arid climate to attenuate solar reflection in 2015-2017, and it concurrently increased soil temperature and fruit-zone high temperature duration and decreased low temperature duration. Inter-row mulch decreased anthocyanins concentrations in grapes in 2015-2016, and consistently inhibited flavonols accumulation in 2015-2017. Correlation analysis between microclimate parameters and flavonoids concentrations reflected the importance of solar reflection, fruit-zone high and low temperature duration, heat accumulation and soil temperature to flavonoids accumulation. Basal leaf removal, a widely applied practice to increase fruit-zone light exposure, was applied to mulch-treated grapevines to investigate if increasing incident light could mitigate the impact of inter-row mulch on flavonoids, and it had limited influence on anthocyanins whereas compensated the loss of flavonols in grapes caused by inter-row mulch. Notably, inter-row mulch wines showed less red and more yellow color than controls because of lower anthocyanins concentrations.

Analysis of the misdiagnosis of 8 adult cases of paragonimiasis with lung masses as the main manifestation in Xishuangbanna, Yunnan.

OBJECTIVE: To summarize the clinical characteristics of adult cases of paragonimiasis with lung masses as the main manifestation in Xishuangbanna, Yunnan Province, analyze the causes of misdiagnosis, and improve the levels of clinical diagnosis and treatment. METHOD: We conducted a retrospective analysis of the clinical data and diagnosis and treatment of 8 adult cases of paragonimiasis with lung masses as the main manifestation that were diagnosed in the Oncology Department of People's hospital of Xishuangbanna Dai Autonomous Prefecture from July 2014 to July 2019. RESULT: All 8 patients were from epidemic paragonimiasis areas and had a confirmed history of consuming uncooked freshwater crabs. The clinical manifestations were mainly fever, dry cough, and chest pain. The disease durations were long, and peripheral blood eosinophil counts were elevated. The cases had been misdiagnosed as pneumonia or pulmonary tuberculosis. After years of anti-inflammatory or anti-tuberculosis treatment, the symptoms had not improved significantly. Patients eventually sought treatment from the oncology department for hemoptysis. Chest computed tomography showed patchy consolidation in the lungs, with nodules, lung masses, and enlarged mediastinal lymph nodes. CONCLUSION: Paragonimiasis is a food-borne parasitic disease. Early clinical manifestations and auxiliary examination results are nonspecific. The parasite most often invades the lungs, and the resulting disease is often misdiagnosed as pneumonia, pulmonary tuberculosis, or lung cancer (Acta Trop 199: 05074, 2019). To avoid misdiagnosis, clinicians should inquire, in detail, about residence history and history of unclean food and exposure to infected water and make an early diagnosis based on the inquired information and imaging examination results. For patients who have been diagnosed with pneumonia or pulmonary tuberculosis and whose symptoms do not improve significantly after anti-inflammatory or anti-tuberculosis treatments, their epidemiological history should be traced to further conduct differential diagnosis and avoid misdiagnosis.

Morphological and Molecular Characterization of Paragonimus Species Isolated from Freshwater Crabs in Southern Yunnan, China.

Paragonimus species are highly prevalent in various regions of China. The study's objective is to isolate and identify Paragonimus from natural habitats and compare the phylogenetic diversity of Paragonimus in southern Yunnan province, China. Metacercariae of Paragonimus was isolated from crabs, and morphologic identification was performed by microscopy. Metacercariae were injected into experimental Paragonimus free Sprague Dawley rats. After 114 days, adult worms and eggs were isolated from multiple organs. Morphologic identification confirmed the initial identification. DNA was extracted from 5 adult worms, and molecular characterization was performed by amplification and sequencing of CO1 and ITS2 regions, followed by phylogenetic analysis. Out of 447 crabs captured, 186 crabs were found to be infected. A total of 4 species of Paragonimus was observed from naturally infected crabs. Paragonimus microrchis (2), Paragonimus heterotremus (1), Paragonimus proliferus (1), and Paragonimus skrjabini (1) were isolated and identified. A total of 32 sequences were downloaded from the National Center for Biotechnology Information, and 5 sequences generated in the study were used for phylogenetic analysis. In the phylogenetic tree of the CO1 gene, Paragonimus proliferus, Paragonimus heterotremus, and Paragonimus skrjabini were clustered with the same species, and the confidence values of their branches were >95%. A congruent phylogenetic relationship was observed with the ITS2 phylogenetic tree. In the phylogenetic tree constructed with the combined dataset of CO1 and ITS2 datasets, Paragonimus proliferus, Paragonimus heterotremus, and Paragonimus skrjabini clustered with the same species, and their branch confidence values were >94%. Paragonimus microrchis clustered with Paragonimus bangkokensis in both datasets. Phylogenetic analysis revealed robustness of the double loci method as against the single-locus method with either CO1 or ITS2 alone. Paragonimus species isolated from the southern Yunnan province, China, was phylogenetically diverse, and the analysis revealed the clustering of multiple species of Paragonimus isolated from different geographic locations.

Influence of attenuated reflected solar radiation from the vineyard floor on volatile compounds in Cabernet Sauvignon grapes and wines of the north foot of Mt. Tianshan.

In this study, fruit-zone microclimate was modified by three treatments, including inter-row mulch (M), the combination of leaf removal applied at the onset of veraison and inter-row mulch (MLR-BV), and the combination of leaf removal applied at complete veraison and inter-row mulch (MLR-EV), in a semi-arid climate in three consecutive years (2015-2017). M decreased fruit-zone reflected solar radiation from vineyard floor and low temperature (10-20 °C) duration, whereas it increased soil temperature and high temperature (> 30 °C) duration. MLR-BV and MLR-EV increased fruit-zone incident photosynthetically active radiation while decreased the duration of 20-25 °C compared to M. Notably, M significantly decreased grape total norisoprenoid concentrations in 2015-2017, and total terpenoid concentrations in 2015-2016. Applying leaf removal applied at the onset of veraison could compensate the decreases of total norisoprenoids and terpenoids caused by M when two treatments were applied together. Besides, M significantly increased grape total C6/C9 compound concentrations, besides, (Z)-3-hexen-1-ol concentrations were significantly higher in grapes of M than those of MLR-BV in 2015-2017. Light exposure and high temperature duration after veraison had strong positive correlations with total norisoprenoids and terpenoids, besides, low temperature duration was positively correlated with total norisoprenoids. In addition, light exposure after veraison had strong negative correlations with total C6/C9 compounds. With respect to the volatile compounds in wines, M significantly decreased the concentrations of isopentanol and ethyl acetate, and the concentrations of ethyl cinnamate, phenylacetaldehyde, phenylethyl alcohol and 3-methylthio-1-propanol were significantly lower in MLR-BV and MLR-EV than in M. The outcome of this study can assist winegrowers to properly adjust vineyard managements to optimize the concentrations of desired volatile compounds in grapes and wines.

Influence of the harvest date on berry compositions and wine profiles of Vitis vinifera L. cv. 'Cabernet Sauvignon' under a semiarid continental climate over two consecutive years.

The ripeness of a grape is critical to berry composition and to the resultant wine. For wineries with a single cultivar occupying an extensive area, the total soluble solid of grapes can range from 22°Brix to 28°Brix. Accordingly, the influence of different harvest dates (ripeness) on berry compositions and on the resultant wine profile was investigated for Vitis vinifera L. cv. 'Cabernet Sauvignon.' Berry dehydration was observed as berry weight and juice yields decreased. Berry anthocyanins were concentrated and methylated anthocyanin levels fluctuated with increasing delays in harvesting. Hexanal and 2-hexenal levels in must decreased significantly as berries ripened. In the resultant wines, 2,3-butanediol levels increased. Wines harvested earlier were lighter, presented lower color intensity (CI) values and higher yellow% levels, and exhibited richer aroma profiles (compounds). Through a principal component analysis and discriminant analysis, the compounds characterizing each harvest date were identified.

MicroRNA-30b regulates insulin sensitivity by targeting SERCA2b in non-alcoholic fatty liver disease.

BACKGROUND & AIMS: Insulin resistance is strongly associated with non-alcoholic fatty liver disease, a chronic, obesity-related liver disease. Increased endoplasmic reticulum (ER) stress plays an important role in the development of insulin resistance. In this study, we investigated the roles of miRNAs in regulating ER stress in the liver of rats with obesity. METHODS: We used miRNA microarray to determine the miRNA expression profiles in the liver of rats fed with a high fat diet (HFD). We used prediction algorithms and luciferase reporter assay to identify the target gene of miRNAs. To overexpress the miRNA miR-30b or inhibit miR-30b rats were injected with lentivirus particles containing PGLV3-miR-30b or PGLV3-miR-30b antimiR through tail vein. Hepatic steatosis was measured using transient elastography in human subjects. RESULTS: Our data showed that miR-30b was markedly up-regulated in the liver of HFD-treated rats. Bioinformatic and in vitro and in vivo studies led us to identify sarco(endo)plasmic reticulum Ca2+ -ATPase 2b (SERCA2b), as a novel target of miR-30b. Overexpression of miR-30b induced ER stress and insulin resistance in rats fed with normal diet, whereas inhibition of miR-30b by miR-30b antimiR suppressed ER stress and insulin resistance in HFD-treated rats. Finally, our data demonstrated that there was a positive correlation between serum miR-30b levels and hepatic steatosis or homoeostasis model assessment of insulin resistance (HOMA-IR) in human subjects. CONCLUSIONS: Our findings suggest that miR-30b represents not only a potential target for the treatment of insulin resistance, but also a non-invasive disease biomarker of NAFLD.

Decreased microRNA-182-5p helps alendronate promote osteoblast proliferation and differentiation in osteoporosis via the Rap1/MAPK pathway.

Osteoporosis (OP) is a serious health problem that contributes to osteoporotic structural damage and bone fragility. MicroRNAs (miRNAs) can exert important functions over bone endocrinology. Therefore, it is of substantial significance to clarify the expression and function of miRNAs in bone endocrine physiology and pathology to improve the potential therapeutic value for metabolism-related bone diseases. We explored the effect of microRNA-182-5p (miR-182-5p) on osteoblast proliferation and differentiation in OP rats after alendronate (ALN) treatment by targeting adenylyl cyclase isoform 6 (ADCY6) through the Rap1/mitogen-activated protein kinase (MAPK) signaling pathway. Rat models of OP were established to observe the effect of ALN on OP, and the expression of miR-182-5p, ADCY6 and the Rap1/MAPK signaling pathway-related genes was determined. To determine the roles of miR-182-5p and ADCY6 in OP after ALN treatment, the relationship between miR-182 and ADCY6 was initially verified. Osteoblasts were subsequently extracted and transfected with a miR-182-5p inhibitor, miR-182-5p mimic, si-ADCY6 and the MAPK signaling pathway inhibitor U0126. Cell proliferation, apoptosis and differentiation were also determined. ALN treatment was able to ease the symptoms of OP. miR-182-5p negatively targeted ADCY6 to inhibit the Rap1/MAPK signaling pathway. Cells transfected with miR-182 inhibitor decreased the expression of ALP, BGP and COL I, which indicated that the down-regulation of miR-182-5p promoted cell differentiation and cell proliferation and inhibited cell apoptosis. In conclusion, the present study shows that down-regulated miR-182-5p promotes the proliferation and differentiation of osteoblasts in OP rats through Rap1/MAPK signaling pathway activation by up-regulating ADCY6, which may represent a novel target for OP treatment.

Effects of Ginsenoside Rb1 on Expressions of Phosphorylation Akt/Phosphorylation mTOR/Phosphorylation PTEN in Artificial Abnormal Hippocampal Microenvironment in Rats.

Artificial abnormal microenvironment caused by microperfusion of L-glutamate (Glu) and Ca2+ in the hippocampus results in neuron damage, which is closely related to cerebral ischemia. Ginsenoside Rb1, a compound from Panax notoginseng, was previously used to counter the artificial abnormal hippocampal environment in a microperfusion model. In addition, while the Akt/mTOR/PTEN signaling pathway has been shown to mediate neuronprotection in cerebral ischemia, whether this pathway is involved in the neuroprotection of ginsenoside Rb1 is unknown. Here SH-SY5Y cells exposed to OGD/R injury in treated with LY294002, ginsenoside Rb1, ginsenoside Rb1+ LY294002. Expressions of phosphorylation (P-)Akt/P-mTOR/P-PTEN (24 h after OGD/R) were detected by Western blotting. Effects were examined via the memory function of rats (by Morris water maze test), morphological changes in pyramidal cell (by histology), and mRNA expression (by qRT-PCR) and phosphorylation (P-) (by Western blotting and immunohistochemical staining) of Akt, P-mTOR, and P-PTEN in the hippocampus. The memory deficit of rats and pyramidal cellular necrosis and apoptosis in the CA1 region of hippocampus after microperfusion of Glu and Ca2+ were dose dependently alleviated by ginsenoside Rb1.Moreover,Western blot showed that ginsenoside Rb1 increased the expressions of P-Akt, P-mTOR and reduced P-PTEN in vivo and vitro. Thus, the potent neuroprotection of ginsenoside Rb1 in artificial abnormal microenvironment is, at least partially, related to the activation of P-AKT/P-mTOR signaling pathway and inhibition of P-PTEN protein.

Intravitreal Bevacizumab Injection Attenuates Diabetic Retinopathy in Adult Rats with Experimentally Induced Diabetes in the Early Stage.

Diabetic retinopathy is the leading cause of blindness, yet its treatment is very limited. Anti-VEGF drug has been widely applied in ocular disease, but its effects on diabetic retinopathy and the underlying mechanism have remained to be fully explored. To elucidate the role of anti-VEGF treatment, we sought to determine the effects of bevacizumab on diabetic neurovascular changes extending from the 3rd to 9th week with induced diabetes in adult rats. The retinal neurovascular changes included increased expression of VEGF, nNOS, iNOS, eNOS, and NO in the course of diabetes progression. In diabetic rats given bevacizumab injection, the ganglion cell loss and alterations of retinal thickness were ameliorated. In this connection, the immunofluorescence labeling of the above biomarkers was noticeably decreased. Along with this, Western blotting confirmed that bevacizumab treatment was associated with a decrease of VEGF, Flk-1, and cAMP response element binding and protein kinase C protein expression. The present results suggest that bevacizumab treatment in the early stage of the retinopathy may ameliorate the lesions of retinopathy, in which VEGF/Flk-1 signaling has been shown here to play an important role.

Panax notoginsenoside saponins Rb1 regulates the expressions of Akt/ mTOR/PTEN signals in the hippocampus after focal cerebral ischemia in rats.

Panax notoginsenoside saponins Rb1 (PNS-Rb1) is an important active ingredient of panax notoginseng for effective treatment of cerebrovascular diseases. However, the mechanism underlying its actions in the state of cerebral ischemia is still unclear. We asked whether the potential neuroprotection of PNS-Rb1 on the brain is due to, at least partially, its modulation of AkT/mTOR/PTEN signalling pathway along with down-regulation of caspase-3 in rats subjected to phototrombic stroke. To test this hypothesis, rats with induced photothrombotic stroke were treated with PNS-Rb1 (applied in three different doses, 25 mg/kg, 50 mg/kg,100 mg/kg, respectively) or saline, while sham operated rats injected with saline were used as the control. Our results indicate that PNS-Rb1 significantly alleviated the morphological lesion concomitant with improvement of cognitive and sensorimotor deficits induced by ischemic stroke. Moreover, immunohistochemistry and Western blot analyses showed that PNS Rb1 in a dose dependent manner increased the expressions of P-Akt, P-mTOR and reduced P-PTEN and caspase-3. The present study suggests that the improvement of cognitive and sensorimotor deficits by PNS-Rb1 is made, at least partially, by the modulation of the Akt/mTOR/PTEN signalling pathway.

Effects of MicroRNA-206 on Osteosarcoma Cell Proliferation, Apoptosis, Migration and Invasion by Targeting ANXA2 Through the AKT Signaling Pathway.

BACKGROUND/AIMS: This study aimed to investigate the mechanism by which microRNA-206 (miR-206) affects the proliferation, apoptosis, migration and invasion of osteosarcoma (OS) cells by targeting ANXA2 via the AKT signaling pathway. METHODS: A total of 132 OS tissues and 120 osteochondroma tissues were examined in this study. The targeting relationship between miR-206 and ANXA2 was verified with a dual-luciferase reporter assay. The miR-206 expression and ANXA2, AKT, PARP, FASN, Survivin, Bax, Mcl-1 and Bcl-1 mRNA and protein expression in the above two groups were examined by qRT-PCR and western blotting. The cultured OS cells were divided into 6 groups: a blank group, negative control (NC) group, miR-206 mimic group, miR-206 inhibitor group, si-ANXA2 group and miR-206 inhibitor + si-ANXA2 group. Cell cycle and apoptosis were assessed by flow cytometry, cell migration was examined with a wound-healing assay, and cell invasion was assessed with a Transwell assay. Pearson correlation analysis was used to determine the correlation between ANXA2 mRNA expression and miR-206 expression in OS. RESULTS: OS tissues exhibited increased mRNA and protein expression of ANXA2, AKT, PARP, FASN, Survivin, Mcl-1 and Bcl-2; decreased miR-206 expression; and decreased Bax mRNA and protein expression. ANXA2 mRNA expression was strongly negatively correlated with miR-206 expression in OS. ANXA2 was found to be a miR-206 target gene. In the miR-206 mimic group and the si-ANXA2 group, the mRNA and protein expression of ANXA2, AKT, PARP, FASN, Survivin, Mcl-1 and Bcl-1 decreased markedly, cell proliferation was inhibited, apoptosis was promoted, higher cell growth in G1 phase and decreased growth in S phase was detected, and decreased cell migration and invasion were observed compared with those in the blank group. CONCLUSION: The current results demonstrate that miR-206 overexpression inhibits OS cell proliferation, migration and invasion and promotes apoptosis through targeting ANXA2 by blocking the AKT signaling pathway.

Insulin-loaded PLGA microspheres for glucose-responsive release.

Porous poly(lactic-co-glycolic acid) (PLGA) microspheres were prepared, loaded with insulin, and then coated in poly(vinyl alcohol) (PVA) and a novel boronic acid-containing copolymer [poly(acrylamide phenyl boronic acid-co-N-vinylcaprolactam); p(AAPBA-co-NVCL)]. Multilayer microspheres were generated using a layer-by-layer approach depositing alternating coats of PVA and p(AAPBA-co-NVCL) on the PLGA surface, with the optimal system found to be that with eight alternating layers of each coating. The resultant material comprised spherical particles with a porous PLGA core and the pores covered in the coating layers. Insulin could successfully be loaded into the particles, with loading capacity and encapsulation efficiencies reaching 2.83 ± 0.15 and 82.6 ± 5.1% respectively, and was found to be present in the amorphous form. The insulin-loaded microspheres could regulate drug release in response to a changing concentration of glucose. In vitro and in vivo toxicology tests demonstrated that they are safe and have high biocompatibility. Using the multilayer microspheres to treat diabetic mice, we found they can effectively control blood sugar levels over at least 18 days, retaining their glucose-sensitive properties during this time. Therefore, the novel multilayer microspheres developed in this work have significant potential as smart drug-delivery systems for the treatment of diabetes.