A complementary method with PFBBr-derivatization based on a GC-EI-MS platform for the simultaneous quantitation of short-, medium- and long-chain fatty acids in murine plasma and feces samples.

Fatty acids (FAs) are essential molecules in all organisms and are involved in various physiological and pathophysiological processes. Pentafluorobenzyl bromide (PFBBr) is commonly used for FA derivatization for gas chromatography-mass spectrometry (GC-MS) quantification by chemical ionization (CI). While CI is the conventional ionization mode for PFBBr derivatization, the electron ionization (EI) source has also demonstrated efficacy in achieving satisfactory analytical performance for the analysis of PFB esters. In this study, we present a novel approach utilizing PFBBr-derivatization on a GC-EI-MS platform to quantitatively analyze a comprehensive range of 44 fatty acids (FAs) spanning from C2 to C24. The method's sensitivity, precision, accuracy, linearity, recovery, and matrix effect were rigorously validated against predetermined acceptance criteria. In comparison to the conventional CI ionization mode, the utilization of PFBBr-derivatization in GC-EI-MS exhibits a wider range of applications and achieves comparable sensitivity levels to the conventional CI platform. By using this method, we successfully quantified 44 FAs in plasma and feces samples from the mice with deoxynivalenol (DON)-induced kidney injury. Among these, the levels of most FA species were increased in the DON-exposure group compared with the control group. The orthogonal partial least squares discriminant analysis (OPLS-DA) of all the tested FAs showed a visual separation of the two groups, indicating DON exposure resulted in a disturbance of the FA profile in mice. These results indicate that the established method by integration of GC-MS with PFBBr derivatization is an efficient approach to quantify the comprehensive FA profile, which includes short-, medium- and long-chain FAs. In addition, our study provides new insights into the mechanism underlying DON exposure-induced kidney injury.

Metabolomics study reveals increased deoxycholic acid contributes to deoxynivalenol-mediated intestinal barrier injury.

AIMS: Deoxynivalenol (DON), namely vomitoxin, is one of the most prevalent fungal toxins in cereal crops worldwide. However, the underlying toxic mechanisms of DON remain largely unknown. MAIN METHODS: DON exposure-caused changes in the murine plasma metabolome and gut microbiome were investigated by an LC-MS/MS-based nontargeted metabolomics approach and sequencing of 16S rRNA in fecal samples, respectively. Cellular models were then used to validate the findings from the metabolomics study. KEY FINDINGS: DON exposure increased intestinal barrier permeability evidenced by its-mediated decrease in colonic Claudin 5 and E-cadherin, as well as increases in colonic Ifn-γ, Cxcl9, Cxcl10, and Cxcr3. Furthermore, DON exposure resulted in a significant increase in murine plasma levels of deoxycholic acid (DCA). Also, DON exposure led to gut microbiota dysbiosis, which was associated with DON exposure-caused increase in plasma DCA. In addition, we found not only DON but also DCA dose-dependently caused a significant increase in the levels of IFN-γ, CXCL9, CXCL10, and/or CXCR3, as well as a significant decrease in the expression levels of Claudin 5 and/or E-cadherin in the human colonic epithelial cells (NCM460). SIGNIFICANCE: DON-mediated increase in DCA contributes to DON-caused intestinal injury. DCA may be a potential therapeutic target for DON enterotoxicity.

An endoparasitoid uses its egg surface proteins to regulate its host immune response.

With proteomic analysis, we identified 379 egg surface proteins from an endoparasitoid, Cotesia chilonis. Proteins containing conserved enzymatic domains constitute a large proportion of egg surface components. Some proteins, such as superoxidase dismutase, homolog of C. rubecula 32-kDa protein, and immunoevasive protein-2A, are classical parasitism factors that have known functions in host immunity regulation. Melanization assays revealed that a novel egg surface protein, C. chilonis egg surface serpin domain-containing protein had the same function as a C. chilonis venom serpin, as both suppressed host melanization in a dose-dependent manner. C. chilonis egg surface serpin domain-containing protein is mainly transcribed in C. chilonis oocytes with follicular cells, and it is located on both the anterior and posterior sides of the mature egg surface. Additionally, we used LC-MS/MS to identify 586 binding proteins sourced from C. suppressalis plasma located on the eggshell surface of C. chilonis, which included some immunity-related proteins. These results not only indicate that C. chilonis uses its egg surface proteins to reduce the immune response of its host but also imply that endoparasitoid egg surface proteins might be a new parasitism factor involved in host immune regulation.

Establishment and characterization of the Bactrocera dorsalis (Diptera: Tephritidae) embryonic cell line QAU-Bd-E-2.

In this study, we successfully established a Bactrocera dorsalis (Diptera: Tephritidae) embryonic cell line, i.e., QAU-Bd-E-2, from the insect eggs. The cells have been stably passaged for more than 60 times in TNM-FH medium with 10% fetal bovine serum (FBS). QAU-Bd-E-2 cells are adherent cells. Most of the cells were round, spindle-shaped, and rod-shaped. Round cells accounted for 82.3%, with a diameter of 13.9 ± 2.6 µm; spindle-shaped cells accounted for 9.8%, with the size of 51.2 ± 11.2 µm × 10.3 ± 3.1 µm; the rod-shaped cells accounted for 7.9%, with the size of 35.2 ± 9.4 µm × 12.0 ± 2.5 µm. The mitochondrial cytochrome oxidase I subunit (CoI) gene from QAU-Bd-E-2 cells was amplified, and the 657 bp fragment had a 100% similarity with the CoI gene of B. dorsalis, suggesting that the cell line was derived from B. dorsalis. The chromosome number of QAU-Bd-E-2 cells was mostly 12, which is the same as the B. dorsalis chromosome number. The cell density of QAU-Bd-E-2 cells reached the maximum (3.4 × 106 cells/mL) at 192 h, and the population doubling time was 31.9 h. Bactrocera dorsalis cripavirus (BdCV) could replicate in QAU-Bd-E-2 cells, suggesting that this cell line could be used for in-depth study of the relationship between virus and host.

MiR-126-3p-Enriched Extracellular Vesicles from Hypoxia-Preconditioned VSC 4.1 Neurons Attenuate Ischaemia-Reperfusion-Induced Pain Hypersensitivity by Regulating the PIK3R2-Mediated Pathway.

Recent evidence suggests that hypoxia preconditioning can alter the microRNA (miRNA) profile of extracellular vesicles (EVs) and has better neuroprotective effects when enriched miRs are delivered to recipients. However, the roles of exosomal miRNAs in regulating ischaemia-reperfusion (IR)-induced pain hypersensitivity are largely unknown. Thus, we isolated EVs from normoxia-conditioned neurons (Nor-VSC EVs) and Hypo-VSC EVs by ultracentrifugation. After the initial screening by a microarray analysis and quantitative RT-PCR (qRT-PCR), miR-126-3p, which was detected as the most altered miR in the Hypo-VSC EVs, was further confirmed by applying GW4869 to inhibit exosomal secretion. Moreover, transfection with a miR-126 mimic obviously increased miR-126-3p expression in Nor-VSC EVs, whereas a miR-126 inhibitor prevented the increase in miR-126-3p in Hypo-VSC EVs. A rat model of pain was established by performing 8-min occlusion of the aorta. Following IR, compared with the Nor-VSC EVs- or antagomir-126-injected rats, the Hypo-VSC EVs-injected rats displayed improved pain hypersensitivity demonstrated as higher PWT and PWL values. Mechanistically, PIK3R2 is a target of miR-126-3p and might be a modulator of the phosphoinositide 3-kinase (PI3K)/Akt pathway as the PIK3R2 and PI3K immunoreactivities in each group were changed in opposite directions. Compared with the controls, higher protein levels of PI3K and phosphorylated Akt but lower levels of phosphorylated nuclear factor-κ B (NF-κB), tumour necrosis factor (TNF)-α and interleukin (IL)-1ß were detected in the spinal cords of the Hypo-VSC EVs-injected rats, and these effects were impaired by an injection of Hypo-VSC EVs combined with antagomir-126. Collectively, the miR-126-3p-enriched Hypo-VSC EVs attenuated IR-induced pain hypersensitivity by restoring miR-126-3p expression in the injured spinal cord and subsequently modulating PIK3R2-mediated PI3K/Akt and NF-κB signalling pathways.

A comparative study of the metabolic effects of LSG and LRYGB in Chinese diabetes patients with BMI<35 kg/m2.

BACKGROUND: The metabolic effects of laparoscopic sleeve gastrectomy (LSG) and laparoscopic Roux-en-Y gastric bypass (LRYGB) in type 2 diabetes (T2D) patients who do not meet National Institutes of Health indications has not been well studied. OBJECTIVES: To compare the effectiveness of LSG and LRYGB in Chinese T2D patients with body mass index (BMI)<35 kg/m2. SETTING: University hospital, China. METHODS: A nonrandomized cohort of patients who underwent LRYGB (n = 64) and LSG (n = 19) were followed up for 3 years and the outcomes (weight loss and remission of diabetes and other metabolic parameters) were compared. Univariate and multivariate analyses were applied to find associated parameters of T2D remission. RESULTS: In total, 5 patients (6%) were lost to follow-up. No significant differences in mean percentage of excess weight loss and BMI were observed between the 2 groups at 2 years. At 3-year follow-up, the LRYGB group had significantly higher percentage of excess weight loss and lower BMI. The total (complete and partial) remission rate achieved with both bariatric procedures was 75.9% at 1 year and 56.4% at 3 years. Surgical safety, diabetes remission, and remission of other obesity-related co-morbidities were comparable between the 2 groups. Patients who achieved complete or partial remission had lower fasting plasma glucose, lower plasma glucose at 2 hours, lower glycated hemoglobin, and higher fasting C peptide than the other patients at baseline. High recurrence rates of hypertension and hyperuricemia were observed at 3 years postoperation. CONCLUSIONS: Both LSG and LRYGB are safe and effective bariatric procedures for T2D in this Chinese population with diabetes and BMI<35 kg/m2.

Laparoscopic sleeve gastrectomy versus Roux-en-Y gastric bypass for morbid obesity: a 1:1 matched cohort study in a Chinese population.

OBJECTIVES: This 1:1 matched cohort study with 3-year follow-up aimed to compare the safety and efficacy of LSG with LRYGB for morbid obesity patients. METHODS: From 2009 to 2013, patients undergoing LRYGB (n = 63) were matched with LSG (n = 63) by gender, age, and body mass index (BMI). Major complications, BMI, percentage of excess weight loss (%EWL), and obesity-related comorbidities after 6, 12, 24, and 36 months were compared. RESULTS: Hospital stay and major complication rates were comparable, but operative time in LSG was significantly shorter (83.2 ± 23.7 vs. 108.3 ± 21.3 min). No significant differences in mean %EWL and BMI were observed at 6, 12, 24 months. At 3-year follow-up, mean %EWL in the LRYGB group was significantly higher than in the LSG group (76.5 ± 9.2% vs. 65.7 ± 10.3%) and, consequently, mean BMI was significantly lower in LRYGB (28.2 ± 1.5 vs. 30.9 ± 2.4 kg/m2). No significant differences in remission of comorbidities were observed at 1- or 3-year follow-up. CONCLUSIONS: Both LRYGB and LSG were safe and effective bariatric procedures in this Chinese population, but LRYGB seemed to be superior to LSG in terms of mid-term weight loss.

Bariatric Surgery in China: How Is This New Concept Going?

BACKGROUND: Obesity has become an epidemic in developing countries including China. The use of bariatric surgery to treat obesity has grown in popularity worldwide, but it is still a new concept in China. This study aims to investigate the trends in bariatric surgery in China. METHODS: An electronic search of the MEDLINE, EMBASE, and Chinese National Knowledge Infrastructure was conducted to select studies for this survey. RESULTS: A total of 7779 bariatric procedures were reported from 2001 to 2015, most of which (89.2 %) were performed in the most recent 5 years. Further, 70.9 % of all procedures were performed to treat obesity and related comorbidities, defined as metabolic surgery. The data showed 89.4 % of all operations were performed laparoscopically. The absolute number of bariatric surgeries increased 148.7 times in the last 5 years compared to the 2001-2005 period. The percentage of laparoscopic Roux-en-Y gastric bypasses performed increased from 0 to 62.2 %, and the percentage of laparoscopic sleeve gastrectomies from 0 to 12.7 %. The percentage of laparoscopic adjustable gastric bands increased dramatically from 0 to 73.3 % in the 2006-2010 period, but it dropped quickly to 12.9 % in the 2011-2015 period. Most operations (66.7 %) were conducted in the East area, which is the most developed economic region in China. There was limited surgical innovation or original research reported in China. CONCLUSIONS: Bariatric surgery is still at an early stage in China, but is now experiencing an explosive growth. A national registry system needs to be established to record and provide precise data.

Invasion Genetics of Woolly Apple Aphid (Hemiptera: Aphididae) in China.

In China, the woolly apple aphid, Eriosoma lanigerum (Hausmann), was first detected as an invasive species during the 1910s to 1930s, restricted to Shandong, Liaoning, and Yunnan Provinces. However, since the 1990s, the pest has spread into many other areas of China. To determine the possible spread routes of the recently established populations, the genetic diversity and genetic structure of 24 populations in 10 provinces were analyzed using eight microsatellite loci. Analyses using STRUCTURE software identified two genetic clusters overall. Three populations from Yunnan and Xinjiang consisted of individuals originating from a single cluster. Nineteen populations from eight northern provinces consisted only of individuals from another cluster, which formed a single large and panmictic population, resembling a distinct "supercolony" in Northern China. The other two populations from Yunnan consisted of individuals from both clusters. The possible routes of spread of the recently established populations of E. lanigerum in China were revealed as follows: 1) the populations in Northern China (including these from Henan, Hebei, Shanxi, Shannxi, Jiangsu, and Gansu) may have been introduced from Shandong or Liaoning Provinces; 2) the populations in Yunnan consisted of an early-established population and a population introduced secondarily from Shandong or neighboring areas, indicating that the population in Yunnan has at least two sources; and 3) the recently established populations of E. lanigerum in Xinjiang might not have been introduced from the "supercolony" in Northern China. Knowledge of these routes of spread is useful for avoiding further dissemination and/or additional introductions.

Serumfree culture of the suspension cell line QB-Tn9-4s of the cabbage looper, Trichoplusia ni, is highly productive for virus replication and recombinant protein expression.

Serumfree cultures of insect cells play an important role in the fields of protein engineering, medicine, and biology. In this paper, the suspension cell line QB-Tn9-4s of Trichoplusia ni (Hübner) (Lepidoptera: Noctuidae) was successfully adapted to serumfree Sf-900 III medium and passaged for 52 generations. The adapted QB-Tn9-4s cells grew faster. Their population doubling time shortened from 27.4 hr in serum-containing medium to 24.1 hr, and their maximal density increased by 1.83-fold, reaching 3.50 ×10(6) cells/mL in serumfree culture in T-flasks. The cells readily adapted to spinner culture, with maximum cell density of 4.40 × 10(6) cells/mL in a spinner flask. Although the infection rate of Autographa californica multiple nucleopolyhedrovirus and production of occlusion bodies (OBs) of the adapted QB-Tn9-4s cells were 91.0% and 85.4 OBs/cell, respectively, similar to those of QB-Tn9-4s cells cultured in serum-containing medium and control BTI-Tn5B1-4 cells, their budded virus titer was 4.97 ×10(7) TCID50/mL, significantly higher than those of the latter two. In addition, the expression levels of ß-galactosidase at six days postinfection and secreted alkaline phosphatase at seven days postinfection in the adapted QB-Tn9-4s cells reached 2.98 ± 0.15×10(4) IU/mL and 3.34 ± 0.13 IU/mL, respectively, significantly higher than those of QB-Tn9-4s and control BTI-Tn5B1-4 cultured in serum-containing media. The above findings establish a foundation for industrial production of virus and recombinant proteins in QB-Tn9-4s serumfree culture.

Population structure of the greenhouse whitefly, Trialeurodes vaporariorum (Westwood), an invasive species from the Americas, 60 years after invading China.

Though the greenhouse whitefly, Trialeurodes vaporariorum (Westwood) (Hemiptera: Aleyrodidae) was introduced into China more than 60 years ago, the genetic diversity and structure of this exotic insect pest and virus vector have not been studied. To investigate the population genetic characteristics of this invasive species and to identify potential invasion routes, the genetic diversity and population structure of 17 collections of T. vaporariorum from nine provinces in China were analyzed using seven microsatellite loci. The results of the analyses indicated that the genetic diversity for the populations examined from the four provinces: Jilin, Ningxia, Guizhou and Qinghai, was lower than the genetic diversity of populations from the five provinces: Yunnan, Shandong, Shanxi, Liaoning, and Gansu. The T. vaporariorum populations analyzed in this study grouped as two distinct genetic clusters based on the analysis using STRUCTURE, whereas, 8 clusters were identified based on the BAPS analysis. Of the 136 genetic distance (Fst) values, 128 (94%) were associated with a significant exact test. However, there was no significant relationship between Fst and geographical distance. These results demonstrate that populations of T. vaporariorum in China exhibit significant genetic differentiation, indicating the likelihood that multiple introductions of T. vaporariorum into China have occurred. Also, the populations collected from the provinces of Jilin, Ningxia, Guizhou and Qinghai appear to represent secondary introductions originating from other Chinese provinces.

Chemical characteristics of normal, woolly apple aphid-damaged, and mechanically damaged twigs of six apple cultivars, measured in autumn wood.

Chemical characteristics of normal, woolly apple aphid-damaged, and mechanically damaged twigs of six apple cultivars: Red Fuji, Golden Delicious, Qinguan, Zhaojin 108, Starkrimson, and Red General, were examined in autumn wood to provide abetter understanding of factors related to cultivar resistance to the woolly apple aphid, Eriosoma lanigerum (Hausmann). Chemical measures examined included soluble sugars, soluble proteins and amino acids, total phenolics, and polyphenol oxidase (that enhances the resistance of plants to insects) and superoxide dismutase, peroxidase, and catalase (that degrade waste products in plants). Soluble sugar, protein, and amino acid contents in normal (undamaged) twigs of Red Fuji, aphid-susceptible cultivar, were higher than in mechanically damaged and aphid-damaged twigs. Total phenolic compounds, an important group of defensive compounds against aphids, increased by 30.5 and 6.0% in mechanically damaged twigs of Qinguan and Zhaojin 108, respectively, and decreased by 21.7 and 16.1% in aphid-damaged twigs of Red Fuji and Red General, respectively. Compared with normal twigs, in aphid-damaged twigs, superoxide dismutase, peroxidase, and polyphenol activity all decreased in Red Fuji. The resistance of some apple cultivars to woolly apple aphid during the growth of autumn shoots was related to several of the physiological indices we monitored. The thin epidermis of callus tissue over healed wounds showed increased susceptibility to the attack by woolly apple aphid. Apple cultivar Qinguan with the highest level of resistance to woolly apple aphid in autumn had increased in amino acid, total phenolic compound levels, and enzyme activity after aphid feeding.

Synthesis and luminescence characterization of cerium doped Lu2O3-Y2O3-La2O3 solid solution transparent ceramics.

Ce3+:(Lu0.7Y0.25La0.05)2O3 transparent ceramics were fabricated with nanopowders and sintered in H2 atmosphere. The spectral properties of Ce3+:(Lu0.7Y0.25La0.05)2O3 transparent ceramics were investigated and the luminescence of Ce3+ in the solid solution of Lu2O3, Y2O3 and La2O3 has been found. The ceramics has high density of 8.10 g/cm(3) and short fluorescence lifetimes of 7.15 ns and 26.92 ns. It is expected to be a good fast response high temperature inorganic scintillating materials.

[Expression vector for the inhibitor of growth-1 gene is constructed and the NLS-GFP fusion protein expresses in MRC-5 cells].

OBJECTIVE: To construct the NLS(ING1)-GFP vector, transfer it into MRC-5 cells and establish a cell model expressing NLS (ING1)-GFP fusion protein. METHODS: Firstly, cDNA fragment of nuclear locating sequence (NLS) of inhibitor of growth-1 gene (ING1) was gained by RT-PCR and inserted into multi-clone site of pEGFP-C1 to construct the NLS (ING1)-GFP expression vector. Then the vector was used to transfect the MRC-5 cells to observe the subcellular signal localization of green fluorescence protein (GFP). RESULTS: We successfully constructed the expressing vector of NLS (ING1)-GFP fusion protein. After transferring the fusion expressing vector into MRC-5 cells, we observed that green fluorescence signal located in the cell nucleus. However, the green fluorescence signal located in the cytoplasm in MRC-5 cells transfected with pEGFP-C1 control only expressing GFP. CONCLUSION: In living cells, physiologically p33 ING1b locates absolutely in nucleus. The p33(ING1b) NLS plays a decisive role in the transporting process of subcellular localization.