Three New Lanostanoids with Anti-HCV Effects from Abies nukiangensis.

Three new lanostane derivatives (1-3) and twelve known triterpenoids (4-15) were isolated from the twigs and leaves of Abies nukiangensis. The structures of the new compounds were elucidated mainly by detailed analysis of their NMR and HR-ESI-MS spectroscopic data. Evaluation of the anti-HCV effects of all isolates showed that 3 exhibited moderate effect with the EC50 value of 11.09 µM.

Risk factors and predictive value of perioperative neurocognitive disorders in elderly patients with gastrointestinal tumors.

BACKGROUND: This study aims to investigate the risk factors of perioperative neurocognitive disorders (PNDs) mainly including postoperative cognitive dysfunction (POCD) in elderly patients with gastrointestinal tumors, and evaluate its predictive value. METHODS: A total of 222 eligible elderly patients (≥65 years) scheduled for elective gastroenterectomy under general anesthesia were enrolled. The cognitive function assessment was carried out 1 day before surgery and 7 days after surgery. Receiver operating characteristic curve analysis was performed to evaluate the predictive value of risk factors for early POCD. The risk factors for POCD were analyzed using univariate and multivariate logistic regression model. RESULTS: Of all the 222 enrolled patients, 91 (41.0%) developed early POCD and 40 (18.0%) were identified as major POCD within 7 days after the surgery. Visual analogue score (VAS, 1st day, resting) ≥4 (OR = 7.618[3.231-17.962], P < 0.001) and alcohol exposure (OR = 2.398[1.174-4.900], P = 0.016) were independent risk factors for early POCD. VAS score (1st, resting) ≥4 (OR = 13.823[4.779-39.981], P < 0.001), preoperative white blood cell (WBC) levels ≥10 × 10*9/L (OR = 5.548[1.128-26.221], P = 0.035), blood loss ≥500 ml (OR = 3.317[1.094-10.059], P = 0.034), history of hypertension (OR = 3.046[1.267-7.322], P = 0.013), and neutrophil-lymphocyte ratio (NLR) ≥2 (OR = 3.261[1.020-10.419], P = 0.046) were independent risk factors for major POCD. Receiver operating characteristic curve analysis indicated that VAS score (1st day, resting) was a significant predictor for major POCD with a cut-off value of 2.68 and an area under the curve of 0.860 (95% confidence interval: 0.801-0.920, P < 0.001). CONCLUSIONS: The risk factors for early POCD after gastroenterectomy included high VAS score (1st day, resting) and alcohol exposure. High VAS score, preoperative WBC levels ≥10 × 10*9/L, blood loss ≥500 ml, NLR ≥2, and history of hypertension were independent risk factors for major POCD. Among them, VAS score was one of the important predictors.

Metformin alleviates inflammatory response in non-alcoholic steatohepatitis by restraining signal transducer and activator of transcription 3-mediated autophagy inhibition in vitro and in vivo.

Autophagy is an intracellular recycling and degradation process for regulating cell survival and drug resistance. Non-alcoholic steatohepatitis (NASH) is becoming a widespread disease in developing countries. However, the role of autophagy in NASH has not yet been fully elucidated. The present study determined that signal transducer and activator of transcription 3 (STAT3), in the inflammation and autophagy regulation, was the key in the progression of NASH. In NASH mouse and cell models, STAT3 mRNA and protein expressions were significantly increased, while the induction of autophagy was radically decreased. Furthermore, the effects of metformin on STAT3 expression level and NASH inflammation were investigated. The current results showed that metformin activated autophagy and decreased the mRNA expressions of inflammatory cytokines, IL-1ß, IL-6, and TNF-α via inhibition of the STAT3 mRNA and protein expression. The siRNA targeting STAT3 activated autophagy and inhibited the NASH inflammatory response by reducing the mRNA expressions of the inflammatory cytokines in vivo and in vitro. The correlation between autophagy and inflammation was also explored. Autophagy induced by metformin attenuated the inflammatory response. This phenomenon of inflammation reduction was partially restored by treatment with the autophagy inhibitor 3-methylindole (3-MA). In conclusion, this study demonstrated that metformin alleviated the inflammatory response in the liver and the hepatocyte of the NASH model via STAT3-mediated autophagy induction. This mechanism provides a strategy for targeting the NASH inflammatory response.

Abinukitrine A, a unique 17,18-cyclolanostane triterpenoid from Abies nukiangensis.

Abinukitrine A (1), a novel triterpenoid, was isolated from Abies nukiangensis. Comprehensive spectroscopic analysis revealed that 1 is the first example of 17,18-cyclolanostane bearing a unique 6/6/6/5/3 ring system. Its absolute configuration was unequivocally assigned by Cu-Kα X-ray single-crystallography. Compound 1 showed a potent anti-hepatitis C virus (HCV) effect.

MiR-134, epigenetically silenced in gliomas, could mitigate the malignant phenotype by targeting KRAS.

Gliomas are characterized by a malignant phenotype with proliferation, cell cycle arrest and invasion. To explore the biological consequences of epigenetically regulated miRNAs, we performed a microarray-based screening (whose expression was affected by 5-AZA treatment) followed by bisulfite sequencing validation. We found that miR-134 as an epigenetically regulated suppressor gene with prognostic value in gliomas. MicroRNA-134 was downregulated in high-grade gliomas, especially in GBM samples. Functional studies in vitro and in vivo in mouse models showed that overexpression of miR-134 was sufficient to reduce cell cycle arrest, cell proliferation and invasion. Target analysis and functional assays correlated the malignant phenotype with miR-134 target gene KRAS, an established upstream regulator of ERK and AKT pathways. Overall, our results highlighted a role for miR-134 in explaining the malignant phenotype of gliomas and suggested its relevance as a target to develop for early diagnostics and therapy.

Overexpression of Transforming Acidic Coiled Coil­Containing Protein 3 Reflects Malignant Characteristics and Poor Prognosis of Glioma.

Gliomas are malignant primary brain tumors with poor prognosis. Recently, research was indicative of a tight connection between tumor malignancy and genetic alterations. Here, we propose an oncogenic implication of transforming acidic coiled-coil-containing protein 3 (TACC3) in gliomas. By comprehensively analyzing the Chinese glioma genome atlas (CGGA) and publicly available data, we demonstrated that TACC3 were overexpressed along with glioma grade and served as an independent negative prognostic biomarker for glioma patients. Functions' annotations and gene sets' enrichment analysis suggested that TACC3 may participate in cell cycle, DNA repair, epithelium-mesenchymal transition and other tumor-related biological processes and molecular pathways. Patients with high TACC3 expression showed CD133⁺ stem cell properties, glioma plasticity and shorter overall survival time under chemo-/radio-therapy. Additionally, a TACC3 associated the miRNA-mRNA network was constructed based on in silico prediction and expression pattern, which provide a foundation for further detection of TACC3-miRNA-mRNA axis function. Collectively, our observations identify TACC3 as an oncogene of tumor malignancy, as well as a prognostic and motoring biomarker for glioma patients.

A bio-inspired synthetic route to the core ring systems of Spiraea atisine-type diterpenoid alkaloids and related diterpenes.

A bio-inspired synthetic strategy for the efficient construction of the structurally complex azapentacyclic ABEFG ring systems of Spiraea atisine-type diterpenoid alkaloids bearing a characteristic internal carbinolamine ether linkage between C(7) and C(20) has been successfully developed. The highly bridged azapentacyclic core structure was constructed rapidly from a readily prepared trans-6,6-bicyclic AB ring precursor through a 14-step sequence. Highlights of the synthesis include a straightforward formal lactone migration from the tricyclic γ-lactone unit of naturally occurring spiramilactone E, and an efficient biomimetic synthesis of the azapentacyclic ABEFG ring systems of spiramines C and D from the corresponding tetracyclic subunits of spiraminol and spiramilactone B.

RAB34 was a progression- and prognosis-associated biomarker in gliomas.

The objective of this study is to explore the expression pattern, prognostic value, and functional role of RAB34 in gliomas. RAB34 messenger RNA (mRNA) expression was evaluated from low grade to high grade in 220 glioma patients of the Chinese Glioma Genome Atlas (CGGA). We therefore analyzed RAB34 mRNA expression in two validated datasets. For detecting the protein expression level of RAB34, another 104 glioma tissues were stained by immunohistochemistry. Gene ontology (GO) analysis and gene set variation analysis (GSVA) were used for functional annotation of RAB34. The mRNA and protein expression levels of RAB34 were both related to glioma grade progression and were inversely correlated with overall survival (OS) in high-grade glioma patients. GO analysis and GSVA showed that RAB34 sets related to migration were significantly enriched in the cases with RAB34 high expression. Pearson correlation analysis identified that genes including MMP-11, HSPB1, IGFBP2, HSPA6, IGFBP5, and MMP19 were positively correlated with RAB34. The expression of RAB34 is related to glioma grade progression and confers a poor prognosis in high-grade glioma patients.

Oxygen glucose deprivation post-conditioning protects cortical neurons against oxygen-glucose deprivation injury: role of HSP70 and inhibition of apoptosis.

In the present study, we examined the effect of oxygen glucose deprivation (OGD) post-conditioning (PostC) on neural cell apoptosis in OGD-PostC model and the protective effect on primary cortical neurons against OGD injury in vitro. Four-h OGD was induced by OGD by using a specialized and humidified chamber. To initiate OGD, culture medium was replaced with de-oxygenated and glucose-free extracellular solution-Locke's medium. After OGD treatment for 4 h, cells were then allowed to recover for 6 h or 20 h. Then lactate dehydrogenase (LDH) release assay, Western blotting and flow cytometry were used to detect cell death, protein levels and apoptotic cells, respectively. For the PostC treatment, three cycles of 15-min OGD, followed by 15 min normal cultivation, were applied immediately after injurious 4-h OGD. Cells were then allowed to recover for 6 h or 20 h, and cell death was assessed by LDH release assay. Apoptotic cells were flow cytometrically evaluated after 4-h OGD, followed by re-oxygenation for 20 h (O4/R20). In addition, Western blotting was used to examine the expression of heat-shock protein 70 (HSP70), Bcl-2 and Bax. The ratio of Bcl-2 expression was (0.44±0.08)% and (0.76±0.10)%, and that of Bax expression was (0.51±0.05)% and (0.39±0.04)%, and that of HSP70 was (0.42±0.031)% and (0.72±0.045)% respectively in OGD group and PostC group. After O4/R6, the rate of neuron death in PostC group and OGD groups was (28.96±3.03)% and (37.02±4.47)%, respectively. Therefore, the PostC treatment could up-regulate the expression of HSP70 and Bcl-2, but down-regulate Bax expression. As compared with OGD group, OGD-induced neuron death and apoptosis were significantly decreased in PostC group (P<0.05). These findings suggest that PostC inhibited OGD-induced neuron death. This neuro-protective effect is likely achieved by anti-apoptotic mechanisms and is associated with over-expression of HSP70.

Covalent organic framework-sodium alginate-Ca2+-polyacrylic acid composite beads for convenient dispersive solid-phase extraction of neonicotinoid insecticides in fruit and vegetables.

Neonicotinoids, the fastest-growing class of insecticides, have posed a multi-media residue problem with adverse effects on environment, biodiversity and human health. Herein, covalent organic framework-sodium alginate-Ca2+-polyacrylic acid composite beads (CACPs), facilely prepared at room temperature, were used in convenient dispersive solid-phase extraction (dSPE) and combined with high-performance liquid chromatography (HPLC) for the detection of five neonicotinoid insecticides (thiamethoxam, acetamiprid, dinotefuran, clothianidin, imidacloprid). CACPs can be completely separated within 1 min without centrifugation. After seven adsorption/desorption cycles, it maintained high extraction efficiencies (>90%). The developed method exhibited a wide linear range (0.01 âˆ¼ 10 µg mL-1), low limits of detection (LODs, 0.0028 âˆ¼ 0.0031 mg kg-1), and good repeatability (RSD ≤ 8.11%, n = 3). Moreover, it was applied to the determination of five neonicotinoids in fruit and vegetables (peach, pear, lettuce, cucumber, tomato), and recoveries ranged from 73.6% to 116.2%.

Analysis of the improved mechanism of Rhodobacter sphaeroides VK-2-3 coenzyme Q10 by reverse metabolic engineering.

Coenzyme Q10 (CoQ10) is an essential medicinal ingredient. In this study, we obtained a high-yielding mutant strain of CoQ10, VK-2-3, by subjecting R. sphaeroides V-0 (V-0) to a 12C6+ heavy ion beam and high-voltage prick electric field treatment. To investigate the mutation mechanism, the complete genomes of VK-2-3 and V-0 were sequenced. Collinearity analysis revealed that the nicotinamide adenine dinucleotide-dependent dehydrogenase (NAD) gene underwent rearrangement in the VK-2-3 genome. The NAD gene was overexpressed and silenced in V-0, and this construct was named RS.NAD and RS.ΔNAD. The results showed that the titers of CoQ10 in the RS.NAD and RS.ΔNAD increased and decreased by 16.00 and 33.92%, respectively, compared to those in V-0, and these differences were significant. Our results revealed the mechanism by which the VK-2-3 CoQ10 yield increases through reverse metabolic engineering, providing insights for genetic breeding and mechanistic analysis.

Application of protective stents in endovascular repair of acute complicated Stanford type B aortic dissections.

PURPOSE: To describe the use of protective stents in the endovascular repair of acute complicated Stanford type B aortic dissections. METHODS: From 2009 to 2011, 33 patients (27 men; mean age 47 years, range 31-73) with acute complicated Stanford type B aortic dissection underwent thoracic endovascular aortic repair (TEVAR) assisted by protective stents. In all cases, the proximal and distal landing zones differed in size by >5 mm, and the primary entry tear was in the proximal descending aorta. A bare self-expanding stent (protective stent) was deployed initially at the intended distal landing site of the primary stent-graft in the true lumen. The intention was that the bare stent would prevent excessive dilation of the distal end of the stent-graft in the vicinity of the entry tear, thus avoiding intimal rupture. RESULTS: Successful stent deployment and sealing of the entry tear was achieved in all patients. The median diameter and length of the protective bare stents was 20.3 mm (range 18-24) and 72.7 mm (range 60-80), respectively, while the corresponding dimensions of the covered stent-grafts were 32.8 mm (range 26-40) and 157.4 mm (range 120-200 mm), respectively. There was no stent twisting, migration, of rupture of the false or true lumen. Computed tomography 1 week postoperatively demonstrated closure of the primary entry tear with thrombosis of the false lumen in all cases. No patients were lost to follow-up, which has ranged from 3 months to 3 years. No late endoleaks or stent complications, such as angulation, dislodgment, persistent leaks, branch obstruction, or stent-graft migration, have been observed, and there has been no chronic progressive true or false lumen dilatation, recurrences, or deaths. CONCLUSION: Adjunctive use of a protective stent when treating acute Stanford type B aortic dissections in which the diameters of the proximal and distal landing zones differ by >5 mm is feasible and safe and provides good short-term outcomes.

Spatial distribution characteristics and prediction of fluorine concentration in groundwater based on driving factors analysis.

Excess fluoride (F-) in groundwater can be hazardous to human health. A total of 360 ground water samples was collected from northern Anhui, China, to study the levels, distribution, and source of F-. And on this basis, predicting the spatial distribution of F- in a wider scale space. The range of F- was 0.1-5.8 mg/L, with a mean value of 1.2 mg/L, and 26.4 % of the samples exceeded the acceptable level of 1.5 mg/L. Moreover, the water-rock interaction (fluorite dissolution) and cation alternate adsorption were considered to be two main driving factors of high F- in groundwater. To further illustrate the spatial effects, the BME-RF model was established by combining the main environmental factors. The spatial distribution of F- was quantitatively predicted, and the response to environmental variables was analyzed. The R2 of BME-RF model reached 0.93, the prediction results showed that the region with 1.0-1.5 mg/L of F- accounts for 47.2 % of the total area. The predicted F- content of nearly 70 % of groundwater in this area has exceeded 1.0 mg/L, which was dominated by Na+ and HCO3- type. The spatial variability of F- in the study area was mainly affected by hydrogeological conditions, and the vertical distribution characteristics were related to the spatial variation of slope, distance from runoff, and hydrochemical types. The results of the study provide new insights into the F- concentration prediction in underground environment, especially in the borehole gap area.

Isolation and Screening of High-Yielding α-Amylase Mutants of Bacillus subtilis by Heavy Ion Mutagenesis.

To improve fermentative production of α-amylase, heavy-ion mutagenesis technology was used to irradiate Bacillus subtilis (B. subtilis) to obtain the high yielding mutants in this study. After continuous cultivation for 12 generations, eight mutants exhibited positive mutation rate with greater H/C. The α-amylase production was stable and obviously exceeded that by the parent strain, which shows that the mutants have a good genetic stability. Among the mutants, the α-amylase activity of B. subtilis KC-180-2 was 72.26 U·mL-1, which was 82.34% higher than that of the original strain. After optimization of fermentation conditions and media, the α-amylase activity of B. subtilis KC-180-2 reached a maximum of 156.83 U·mL-1 at 36 h in a bioreactor. In addition, the optimized fermentation temperature of B. subtilis KC-180-2 was increased to 49℃, indicating B. subtilis KC-180-2 possesses high-temperature resistance, which has great application prospects for industrial fermentation for α-amylase production.

Perioperative approach in the surgical management of carotid body tumors.

BACKGROUND: Now, surgical resection still remains the gold standard for the treatment of carotid body tumors (CBTs). Although advances in surgical techniques and the introduction of sensitive imaging modalities have significantly reduced mortality, the incidence of perioperative neurovascular complications, especially cranial nerve deficit and intraoperative hemorrhage, remains considerable. To solve these problems, preoperative embolization has been suggested; the reported benefits of preoperative embolization performed <48 hours before surgery include a reduction in tumor size, decreased blood loss, and improved visualization, theoretically reducing neurologic morbidity by lessening the risk of stroke and damage to cranial nerves. The purpose of this study was to review our experience in the surgical management of CBTs with preoperative embolization and evaluate the outcomes and complications according to the Shamblin classification. METHODS: Thirty-two patients who had been diagnosed with and surgically treated for CBTs were enrolled from January 2005 till July 2010. All perioperative scans were evaluated by computed tomography angiography. We reviewed patient demographics, radiographic findings, and surgical outcomes collected from medical records. RESULTS: Thirty-two patients underwent surgical excision without mortality. Angiography with selective preoperative tumor embolization was performed on 21 patients. The median blood loss, operation time, and hospital stay for these patients were significantly reduced compared with those without embolization. There were no recurrences or delayed complications at the median follow-up of 20 months. CONCLUSION: Embolization as an adjunctive tool was beneficial for CBT surgery outcomes. Embolization should only be undertaken in those vessels that can be subselectively catheterized and determined not to allow free reflux of contrast medium into the internal carotid artery. Tumor embolization was performed on patients with Cook detachable coils, which are highly effective for supply artery closure if properly selected, and complications can be minimized by proper selection and positioning of the coil. Operation within 48 hours after embolization is recommended to minimize revascularization edema or a local inflammatory response.

New monoterpenes, diterpenes, and lignans from Abies recurvata.

From the aerial part of Abies recurvata, 62 miscellaneous chemical constituents were isolated including 6 new and 56 known ones. The new compounds comprised three monoterpenes, two diterpenes, and one lignan. Their chemical structures were characterized on the basis of various spectroscopic techniques. Dihydrodehydrodiconiferyl alcohol (35) showed the strongest inhibitory effects against lipopolysaccharide-induced nitric oxide production in RAW 264.7 cells with an IC50 value of 66.4 µM.

Overexpression of enzymes in glycolysis and energy metabolic pathways to enhance coenzyme Q10 production in Rhodobacter sphaeroides VK-2-3.

We subjected the components of the glycolysis and energy metabolism pathways of Rhodobacter sphaeroides (R. sphaeroides) to metabolic engineering to improve the titer and yield of coenzyme Q10 (CoQ10). Phosphofructokinase (PFK), cyclic adenylate-dependent protein kinase (PKAC), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and adenosine triphosphate hydrolase (KdpC) were overexpressed in R. sphaeroides VK-2-3 (VK-2-3). The strains were labeled R. sphaeroides PFK (RS.PFK), RS.PKAC, RS.PFK-PKAC, RS.KdpC, RS.GAPDH, and RS.KdpC-GAPDH. Results showed that the CoQ10 titers of RS.PFK, RS.PKAC, and RS.PFK-PKAC were 300.96 ± 0.87, 405.94 ± 4.77, and 379.94 ± 0.42 mg/l, respectively. The CoQ10 titers of RS.PFK and VK-2-3 were not significantly different; however, those for RS.PKAC and RS.PFK-PKAC were 13 and 6% higher than that of VK-2-3, respectively. Further, the titers of RS.KdpC, RS.GAPDH, and RS.KdpC-GAPDH were 360.17 ± 0.39, 409.79 ± 0.76, and 359.87 ± 1.14 mg/l, respectively. The titers of RS.KdpC and RS.KdpC-GAPDH were not significantly different from that for VK-2-3, whereas that for RS.GAPDH was 14% higher than that of VK-2-3. Finally, when the cultures of RS.GAPDH and VK-2-3 were scaled up in 5-L fermenters, the CoQ10 titers and RS.GAPDH yields increased by 44.3 and 37.8%, respectively, compared with VK-2-3.To the best of our knowledge, the glycolysis pathway of R. sphaeroides was studied for the first time in this study. We genetically modified the components of the energy metabolism pathway to obtain the strain with high yield of CoQ10 mutant RS.GAPDH. The findings of this study can serve as a basis for future studies involving metabolic engineering of CoQ10-producing strains.

[Construction and application of lentivirus overexpression vector with two labeling genes fused with CopGFP and PuroR].

Objective: To construct the lentivirus overexpression vector with two label genes fused with CopGFP and PuroR and to detect the emission of green fluorescence as well as resistance to puromycin in liver cancer cells infected with lentivirus packaged with the above vector. Methods: Firstly, two fragments containing copGFP and PuroR coding sequences were amplified from pCDH-CMV-MCS-copGFP and pLKO.1 respectively; secondly, the two amplified regions were fused with each other by recombinant PCR; thirdly, the fusion DNA fragment was cut and inserted into pCDH-CMV-MCS-copGFP vector, which was linearized with the same restriction endonuclease as used to digest fusion DNA fragment: BamH Ⅰ and Sal Ⅰ. The fusion region in the constructed vector was confirmed by DNA sequencing. The checked vector was co-transfected with package assistant plasmids, namely PLP1, PLP2 and VSVG into in 293T cells and the culture supernatant was subjected to centrifuge and infect liver cancer MHCC97H cells, which were then used to detect their resistance to puromycin (infected cells were treated with 1 mg/ml puromycin for 7 days after infection) and to observe green fluorescence emission in microscope. To determine its efficiency in expressing foreign target protein, the Sp1 coding region was inserted into the MCS sites of the vector, and Sp1 mRNA and protein expression levels were compared with the vehicle vector by RT-qPCR and Western blot. Results: The lentivirus overexpression vector with two label genes fused with CopGFP and PuroR was successfully constructed, and the liver cancer cells infected with lentivirus packaged with the vector expressing two labeling genes fused with CopGFP and PuroRshowed both emission of green fluorescence and resistance to puromycin simultaneously, while cells containing with the vector inserted with Sp1 coding region improved Sp1 mRNA level with 3.3 fold and protein level with 2.2 fold higher in comparison with cells containing the vehicle vector (P＜0.01). Conclusion: The fused label genes consisting of copGFP and PuroR are correctly cloned into the lentivirus vector and confer cells with the ability to emission of green fluorescence and resistance to puromycin, besides, the vector may promote the expression of the target gene with long coding sequence.

Immunologic changes during Pandemic (H1N1) 2009, China.

We analyzed changes in immunologic values over time for 28 hospitalized patients with pandemic (H1N1) 2009. Levels of interleukin-6, interferon-y, and interleukin-10 increased 1 day after illness onset and then decreased to baseline levels. Levels of virus-specific antibody were undetectable 1 day after illness onset and peaked 36 days later.