Characterization of a novel anti-PVRIG antibody with Fc-competent function that exerts strong antitumor effects via NK activation in preclinical models.

Poliovirus receptor-related immunoglobulin domain-containing protein, or PVRIG, is a newly discovered immune checkpoint that has emerged as a promising target for cancer immunotherapy. It is primarily expressed on activated T and natural killer (NK) cells, and once engaged with its ligand, PVRL2, it induces inhibitory signaling in T cells, thereby promoting the functional exhaustion of tumor-infiltrating lymphocytes (TILs). Here, we characterized IBI352g4a, a novel humanized anti-PVRIG antibody with Fc-competent function, explored the mechanism of its antitumor activity in preclinical models, and systemically evaluated the contribution of FcrR engagement to PVRIG blockade-induced antitumor activity. IBI352g4a binds to the extracellular domain of human PVRIG with high affinity (Kd = 0.53 nM) and specificity, and fully blocks the interaction between PVRIG and its ligand PVRL2. Unlike other immune checkpoints, IBI352g4a significantly induced NK cell activation and degranulation, but had a minimal effect on T-cell activation in in vitro functional assays. IBI352g4a induced strong antitumor effect in several preclinic models, through in vivo mechanism analysis we found that both NK and T cells contribute to the antitumor effect, but NK cells play predominant roles. Specifically, a single dose of IBI352g4a induced significant NK cell activation in TILs, but T-cell activation was observed only after the second dose. Moreover, the Fc effector function is critical for both NK cell activation and treatment efficacy in vitro and in vivo. Our study, for the first time, demonstrates that both NK activation and FcrR engagement are required for antitumor efficacy induced by PVRIG blockade.

Restoring long-lasting midface volume in the Asian face with a hyaluronic acid filler: A randomized controlled multicenter study.

BACKGROUND: Hyaluronic acid (HA) filler treatment is a minimally-invasive alternative to surgery to volumize the cheeks. HAVOL (Restylane® Volyme) is a flexible HA filler suited to contouring and volumizing the midface. METHODS: This randomized, evaluator-blinded, no-treatment controlled study evaluated effectiveness and safety of HAVOL for correction of midface volume deficit and midface contour deficiency in Chinese subjects. In total 111 subjects were randomized to HAVOL and 37 to no treatment (control). The primary endpoint was response, on the blinded evaluator-assessed Medicis Midface Volume Scale (MMVS), at 6 months after last injection for the treatment group and 6 months after randomization for controls, where response was defined as ≥1-point improvement from baseline on both sides of the face. RESULTS: HAVOL was superior to no treatment at 6 months, meeting the primary objective: 76% versus 8% MMVS responders, a difference of 68% (CI: 55.7%-79.4%, p < 0.0001). These effects were sustained in 51% at 12 months after last injection. A majority (≥96%) had improved aesthetic appearance of midface fullness at Month 1 (using the Global Aesthetic Improvement Scale [GAIS]), effects which remained in ≥80% up to 12 months. Volume change captured by 3D photography increased after 1 month to 3.6 mL (close to the total injected volume of 3.4 mL), and remained stable through 12 months. Over 97% reported satisfaction with results after treatment with HAVOL. Additionally, HAVOL was well tolerated, with no unanticipated related adverse events. CONCLUSIONS: This study showed that HAVOL is effective and well tolerated for midface treatment in a Chinese population.

Determination of acetochlor by UPLC-MS3 in cells and its application to a cellular pharmacokinetic study.

The aim of this work was to develop a fast, simple, and reliable UPLC-MS3 method for the sensitive detection of acetochlor in biological samples. In MS3 mode, the ion transition m/z 270.1 â†’ 224.1→148.1 was chosen for quantification with butachlor as the internal standard. In the UPLC system, separation was performed on a UPLC column (2.1 × 50 mm ID, 1.7 µm) with 0.1 % FA in water and acetonitrile as mobile phases. After simple protein precipitation via acetonitrile, the method was well validated with good linearity (0.5-20 ng/mL, r > 0.995), accuracy (-3.70 %-2.98 %), and precision (<15 %). The selectivity and sensitivity were improved obviously in MS3 mode than that in MRM mode. The developed UPLC-MS3 method was successfully applied to the cellular pharmacokinetics study of acetochlor in MCF-7 cells.

Ultra-high-performance liquid chromatography with tandem mass spectrometry method for cellular toxicity and pharmacokinetic study of PEG1K polymers.

Polyethylene glycol (PEG) is one of the most commonly used polymers in drug delivery systems. The investigation of the pharmacokinetic behavior of PEG is important for revealing the toxicity and efficiency of PEG-related Nano-drug delivery systems. A high through-put and selective ultra-high-performance liquid chromatography with tandem mass spectrometry (UHPLC-MS/MS) method coupled with collision-induced dissociation (CID) in source technique was developed and validated to determine PEG1K polymers in cellular samples in this study. The countless precursor ions of PEG1K are dissociated in the source to generate numerous product ions which have different numbers of subunits. The transition of [M+H]+ precursor ions → product ions at m/z 177.1 (four subunits)→89.1 (two subunits) was selected to determine PEG1K due to its high sensitivity. The UHPLC-MS/MS method coupled with CID in the source showed good linearity over the range of 0.1-10 µg/mL. Intra-day and inter-day accuracies and precisions of the assay were all within ± 12.39%. The assay was successfully applied to a cellular pharmacokinetic study of PEG1K in human breast cancer cells. The cytotoxicity of PEG1K polymers was also studied and the results indicated that the cytotoxicity of PEG1K was not significant in the range of 5-1200 µg/mL.

Ultrafast and ultrastable FeSe2embedded in nitrogen-doped carbon nanofibers anode for sodium-ion half/full batteries.

Transition metal selenides are considered as promising anode materials for fast-charging sodium-ion batteries due to their high theoretical specific capacity. However, the low intrinsic conductivity, particle aggregation, and large volume expansion problems can severely inhibit the high-rate and long-cycle performance of the electrode. Herein, FeSe2nanoparticles embedded in nitrogen-doped carbon nanofibers (FeSe2@NCF) have been synthesized using the electrospinning and selenization process, which can alleviate the volume expansion and particle aggregation during the sodiation/desodiation and improve the electrical conductivity of the electrode. The FeSe2@NCF electrode delivers the outstanding specific capacity of 222.3 mAh g-1at a fast current density of 50 A g-1and 262.1 mAh g-1at 10 A g-1with the 87.8% capacity retention after 5000 cycles. Furthermore, the Na-ion full cells assembled with pre-sodiated FeSe2@NCF as anode and Na3V2(PO4)3/C as cathode exhibit the reversible specific capacity of 117.6 mAh g-1at 5 A g-1with the 84.3% capacity retention after 1000 cycles. This work provides a promising way for the conversion-based metal selenides for the applications as fast-charging sodium-ion battery anode.

Quantification of human serum albumin by combining chymotrypsin/trypsin digestion coupled with LC-MS/MS technique.

The quantification of albumin is important in clinical medicine because the concentration of albumin in biological fluids is closely related to human health. In this study, we developed a highly selective and robust assay to determine human serum albumin (HSA) in human plasma by combining chymotrypsin/trypsin digestion coupled with targeted LC-MS/MS technique. Human plasma samples were denatured, reduced, alkylated, and digested with both chymotrypsin and trypsin to generate surrogate peptides. A unique chymotryptic peptide (NAETF) arising from human serum albumin was finally selected for targeted LC-MS/MS detection and quantification. Numerous parameters related to the targeted LC-MS/MS assay were evaluated, including lower limit of quantitation (LLOQ), linearity range, enzyme digestion efficiency, accuracy and precision. The LC-MS/MS assay was linear in the concentration range 0.05-1 mg/mL with intra-day and inter-day precision <10.2% and accuracy ranging from -3.94% to 4.89%. The assay was successfully applied to determine HSA in 148 human plasma samples.

The Effect of Trans-Sutural Distraction Osteogenesis on Nasal Bone, Nasal Septum, and Nasal Airway in the Treatment for Midfacial Hypoplasia in Growing Patients.

The purposes of this study were to analyze the effect of trans-sutural distraction osteogenesis (TSDO) on nasal bone, nasal septum, and nasal airway in the treatment of midfacial hypoplasia. A total of 29 growing patients with midfacial hypoplasia who underwent TSDO by a single surgeon were enrolled. The 3-dimensional measurement of nasal bone and nasal septum changes was performed using computed tomography (CT) images obtained preoperatively (T0) and postoperatively (T1). One patient was selected to establish 3-dimensional finite element models to simulate the characteristics of nasal airflow field before and after traction. After traction, the nasal bone moved forward significantly ( P <0.01). The septal deviation angle was lower than that before traction (14.43±4.70 versus 16.86 ±4.59 degrees) ( P <0.01). The length of the anterior and posterior margin of the vomer increased by 21.4% ( P <0.01) and 27.6% ( P <0.01), respectively, after TSDO. The length of the posterior margin of the perpendicular plate of ethmoid increased ( P <0.05). The length of the posterior inferior and the posterior superior margin of the nasal septum cartilage increased ( P <0.01) after traction. The cross-sectional area of nasal airway on the deviated side of nasal septum increased by 23.0% after traction ( P <0.05). The analysis of nasal airflow field showed that the pressure and velocity of nasal airflow and the nasal resistance decreased. In conclusion, TSDO can promote the growth of the midface, especially nasal septum, and increase the nasal space. Furthermore, TSDO is conductive to improve nasal septum deviation and decrease nasal airway resistance.

Unraveling the in vivo biological fate of mPEG2000-PDLLA2500-COOH diblock copolymers by LC-MS/MS based on CID in source technique.

Methoxy poly (ethylene glycol)-poly(D, L-lactic acid) (mPEG-PDLLA) is a biocompatible and amphiphilic diblock copolymer composed of a hydrophilic poly(ethylene glycol) block and a hydrophobic poly(D, L-lactic acid) block, which can self-assemble into micelles in aqueous solution. It is one of the most widely used diblock copolymers for drug delivery, drug solubilization and drug encapsulation. Fully characterizing the in vivo fate of mPEG-PDLLA diblock copolymers is important to promote the further development of polymer-based nanocarrier drug delivery systems. However, to date, a bioanalysis assay for simultaneous quantification of mPEG-PDLLA and mPEG has not been reported. In this study, we developed such a novel LC-MS/MS assay based on CID in source technique and used it to study the multiple-dose pharmacokinetic, tissue distribution and excretion of mPEG2000-PDLLA2500-COOH and mPEG2000 in rat after intravenous administration. The results indicate that mPEG2000-PDLLA2500-COOH and mPEG2000 are mainly distributed to the liver, lung, spleen and kidney after intravenous administration. mPEG2000-PDLLA2500-COOH is mostly excreted via the renal route in the form of mPEG2000. Overall, the results of this study provide a comprehensive and clear picture of the in vivo fate of mPEG2000-PDLLA2500-COOH which will be useful in evaluating the efficiency and safety of polymer-based nanocarrier drug delivery systems.

Development and validation of an LC-MS/MS assay with multiple stage fragmentation for the quantification of alachlor in McF-7 cells.

Alachlor is one of the most widely used herbicides and can also be a carcinogenic compound. It is of great significance to establish a sensitive analytical method for the determination of alachlor in the environment and organisms. In this study, a high-performance liquid chromatography tandem mass spectrometry cubed (LC/MS3) method was developed and validated to quantify alachlor in human breast cancer cells (McF-7 cells). The cell samples were processed by simple protein precipitation with acetonitrile, then the analytes were separated on a Waters AcQuity® UPLC BEH (2.1 × 50 mm I.D, 1.7 µm) column using the gradient elution with solvent A (0.1 % formic acid) and solvent B (acetonitrile) at a flow rate of 0.5 mL/min. MS3 detection in positive ion mode was used to detect the analytes. The MRM3 transitions at m/z 270.1 â†’ 238.0 â†’ 162.1 and 312.2 â†’ 238.1 â†’ 147.2 were used to determine alachlor and butachlor, respectively. The run time for each sample was only 4 min. This method was validated for various parameters including accuracy, precision, selectivity, linearity, lower limit of quantitation (LLOQ), etc. The LC/MS3 assay was linear in the concentration range 0.5-50 ng/mL (R2 ≥ 0.995). For all concentrations, the precision is < 9.49 %, and the intra-day and intra-day accuracy is < 13.05 %. Cytotoxic potential of alachlor against McF-7 cell lines was measured by MTT method after 48 h of incubation. For alachlor, half maximal inhibitory concentration (IC50) on McF-7 cells was 87.95 µg/mL. This method was successfully applied to cellular pharmacokinetic study of alachlor in McF-7 cells after administration with a dose of 20 µg/mL.

PI3K/AKT pathway promotes keloid fibroblasts proliferation by enhancing glycolysis under hypoxia.

Our previous study demonstrated altered glucose metabolism and enhanced phosphorylation of the PI3K/AKT pathway in keloid fibroblasts (KFb) under hypoxic conditions. However, whether the PI3K/AKT pathway influences KFb cell function by regulating glucose metabolism under hypoxic conditions remains unclear. Here, we show that when PI3K/AKT pathway was inactivated with LY294002, the protein expression of glycolytic enzymes decreased, while the amount of mitochondria and mitochondrial membrane potential increased. The key parameters of extracellular acidification rate markedly diminished, and those of oxygen consumption rate significantly increased after inhibition of the PI3K/AKT pathway. When the PI3K/AKT pathway was suppressed, the levels of reactive oxygen species (ROS) and mitochondrial ROS (mitoROS) were significantly increased. Meanwhile, cell proliferation, migration and invasion were inhibited, and apoptosis was increased when the PI3K/AKT pathway was blocked. Additionally, cell proliferation was compromised when KFb were treated with both SC79 (an activator of the PI3K/AKT pathway) and 2-deoxy-d-glucose (an inhibitor of glycolysis), compared with the SC79 group. Moreover, a positive feedback mechanism was demonstrated between the PI3K/AKT pathway and hypoxia-inducible factor-1α (HIF-1α). Our data collectively demonstrated that the PI3K/AKT pathway promotes proliferation and inhibits apoptosis in KFb under hypoxia by regulating glycolysis, indicating that the PI3K/AKT signalling pathway could be a therapeutic target for keloids.

Cellular toxicity and pharmacokinetic study of butachlor by ultra-performance liquid chromatography-tandem mass spectrometry based on tandem mass spectrometry cubed technique.

Butachlor is an aromatic amide compound that plays a role as a herbicide, a xenobiotic, and an environmental contaminant. The aim of this work was to develop a highly selective and sensitive ultra-performance liquid chromatography-tandem mass spectrometry method based on the tandem mass spectrometry cubed technique to determine butachlor in a biological matrix. Butachlor and internal standard acetochlor were separated on a Waters Acquity ultra-performance liquid chromatography BEH C18 column (2.1 × 50 mm, 1.7 µm) with gradient elution using 0.1% formic acid aqueous solution (A) and acetonitrile (B) as mobile phases. The transitions selected for tandem mass spectrometry cubed quantitative analysis in positive ion mode were: for butachlor, mass-to-charge ratio 312.2→238.1→162.1; for acetochlor, mass-to-charge ratio 270.1→224.0→148.1. The total running time for each sample was 5.5 min. The ultra-performance liquid chromatography-tandem mass spectrometry cubed method showed a linear relationship (R2 ≥ 0.995) in the concentration range of 0.5-100 ng/ml. The intra and interday accuracies are within the range of -10.6%-4.3% and precisions are between 4.48% and 13.14%. The novelty of the method is the use of tandem mass spectrometry cubed scanning mode, which improves selectivity and sensitivity. The results indicated that butachlor was cellular toxic. The safety of butachlor should be considered when it is used as a herbicide.

Design, synthesis, biological evaluation and molecular docking study of 2,4-diarylimidazoles and 2,4-bis(benzyloxy)-5-arylpyrimidines as novel HSP90 N-terminal inhibitors.

The molecular chaperone HSP90 plays an essential role in cancer occurrence and development. Therefore, it is an important target for the development of anticancer drugs. 1,3-Dibenzyl-2-aryl imidazolidine (8) is a previously reported inhibitor of HSP90; however, its anticancer activity is poor. In this work, chemical modification of 8 led to the discovery of 2,4-diarylimidazoles and 2,4-bis(benzyloxy)-5-arylpyrimidines as two types of novel HSP90 N-terminal inhibitors. 16l and 22k exhibited antiproliferative activity against multiple breast cancer cell lines with IC50 values at the low micromolar level. 16l and 22k induced significant degradation of the client proteins AKT and ERK and a lower level of the heat shock response in comparison with tanespimycin (17-AAG). 22k exhibited a strong affinity for the HSP90α N-terminus with an IC50 value of 0.21 µM. A molecular docking study revealed that 16l and 22k successfully bind to the geldanamycin binding site at the N-terminus of HSP90α.

A facile synthesis of CuSe nanosheets for high-performance sodium-ion hybrid capacitors.

Due to the low price and abundant reserves of sodium resources, sodium-ion batteries have become the main candidate for the next generation of energy storage equipment, particularly for large-scale grid storage and low-speed electric vehicles. Transition metal selenides have attracted considerable attention because of their high reversible capacity, superior electrical conductivity and versatile structures. In this study, two-dimensional CuSe nanosheets are synthesized via a simple hydrothermal reaction. When acting as an electrode material for sodium-ion batteries, the CuSe electrode exhibits an initial coulombic efficiency of 96.7% at a current density of 0.1 A g-1 and a specific capacity of 330 mA h g-1 after 100 operation cycles, as well as retains a specific capacity of 211 mA h g-1 even at a high current density of 10 A g-1. Moreover, the anode delivers a specific capacity of 236 mA h g-1 after 3300 cycles at 5 A g-1 with a capacity retention of 91.2%. In sodium-ion hybrid capacitors (SHICs) with the two-dimensional CuSe nanosheets and Ti3C2T x MXene as the negative and positive materials, respectively, the nanosheets without any pre-sodiation present a lifespan of up to 2000 cycles at 2 A g-1 and a capacity retention of about 77.7%.

Quantitative analysis of polypropylene glycol polymers by liquid chromatography tandem mass spectrometry based on collision induced dissociation technique.

Polypropylene glycol (PPG) is a commonly used synthetic polymer in many fields. Investigating the toxicity and pharmacokinetic behavior of PPG polymers is necessary and important for evaluating their safety in medicine and daily cosmetics. In this study, PPG425, PPG1K and PPG2K were selected as the target polymers for cytotoxicity and cellular pharmacokinetics study of PPG polymers. Structural diversity and polydisperse molecular weights (MWs) are significant challenges for quantification of PPG polymers by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Collision induced dissociation in source or collision cell generated a series of PPG-related product ions at m/z 59.0, 117.1, 175.1, 233.2, 291.2, 349.3, 407.2, 465.3 and 523.5 corresponding to fragments containing 1, 2, 3, 4, 5, 6, 7, 8, 9 repeating propylene oxide subunits. PPG425 was determined by the sum of the MRM acquisitions used the transitions [M+H]+1 precursor ions â†’ product ions. PPG1K and PPG2K were determined by the MRM acquisitions used the transitions [M+H]+1 precursor ions â†’ product ions at m/z 233.2(four subunits)→59.0(one subunit). Based on the collision induced disassociation technique and structural specific product ions, pharmacokinetic studies of PEG425, PPG1K and PPG2K were successfully conducted in McF-7 cells. The experimental results revealed that PPG polymers are not biologically inert and they can enter into McF-7 cells. The safety of PPG polymers should be considered when they are used as pharmaceutical or cosmetic excipients.

Ultrafast and high-throughput quantitative analysis of carbamazepine in human plasma by direct analysis in real time tandem mass spectrometry coupled with solid phase extraction to eliminate matrix effects.

Therapeutic drug monitoring of carbamazepine is necessary for its clinical application with the purpose to reach therapeutic concentration and reduce the risk of concentration-dependent toxicity. An ultrafast analytical assay for quantification of carbamazepine in human plasma was developed and validated based on direct analysis in real time tandem mass spectrometry (DART-MS/MS). After reversed phase solid phase extraction with Waters Oasis HLB, carbamazepine and internal standard carbamazepine-D2N15 were monitored by positive ion mode followed by multiple reaction monitoring (MRM) of the transitions at m/z 237.1→194.0 and 240.1→196.2, respectively. The DART-MS/MS method is ultrafast and high-throughput and the analytical time for each sample is only 0.4 min. The assay was linear in the concentration range 0.50-30 µg/mL and intra- and inter-day accuracies were within ± 15% and trueness were < 13.9% at all concentrations. The method was successfully applied to therapeutic drug monitoring of carbamazepine in human plasma.

Strategy of Virtual Screening based Discovery of HSP90 C-terminal Inhibitors and Network Pharmacological Analysis.

BACKGROUND: HSP90 has been considered an important anticancer target for several decades, but traditional HSP90 N-terminal inhibitors often suffered from organ toxicity and/or drug resistance. METHODS: The development of HSP90 C-terminal inhibitors represents a reliable alternative strategy. In view of rare examples of structure-based identification of HSP90 C-terminal inhibitors, we report a virtual screening based strategy for the discovery of HSP90 C-terminal inhibitors as anticancer agents from natural products. RESULTS & DISCUSSION: 13 chemical ingredients from licorice were identified as possible HSP90 inhibitors and 3 of them have been reported as anticancer agents. The binding modes towards HSP90 C-terminus were predicted by molecular docking and refined by molecular dynamics simulation. CONCLUSION: Further network pharmacological analysis predicted overall possible targets involved in the pathways in cancer and revealed that 8 molecules possibly interact with HSP90. A structure based virtual screening strategy was established for the discovery of HSP90 Cterminal inhibitors.

Effects of a blend of essential oils, medium-chain fatty acids, and a toxin-adsorbing mineral on diarrhea and gut microbiome of weanling pigs experimentally infected with a pathogenic Escherichia coli.

A proprietary antimicrobial feed additive comprised of essential oils, medium-chain fatty acids, and a toxin-adsorbing mineral showed promising bacteriostatic and bactericidal effects in vitro. This study investigated the impacts of supplementing this blend on growth, gut microbiome, and enteric disease resilience in weaned pigs experimentally challenged with an enterotoxigenic Escherichia coli (ETEC). Thirty-six weanling pigs (6.88 ±â€…0.30 kg body weight) blocked by weight and gender were assigned to one of three dietary treatments: control or dietary supplementation with 0.25% or 0.50% of the antimicrobial blend. This study lasted 28 d with 7 d before and 21 d after the first ETEC inoculation (day 0). All pigs were orally inoculated with 1010 CFU F18 + ETEC/3-mL dose for 3 consecutive days. Growth performance data and diarrhea scores were recorded throughout the experiment. Fecal samples collected on days -7, 0, 7, and 21 post first inoculation (PI), and ileal digesta and mucosal tissue collected on day 21 PI were further analyzed for gut microbiome using 16S rRNA sequencing. All data, except for frequency of diarrhea and gut microbiome, were analyzed by ANOVA using the PROC MIXED of SAS. The chi-square test was used for analyzing frequency of diarrhea. Gut microbiome data were analyzed using QIIME2 and visualized using the R program. Dietary supplementation of 0.25% or 0.5% of the antimicrobial blend increased (P < 0.05) feed efficiency on days 14 to 21 PI of ETEC and reduced (P < 0.05) frequency of diarrhea during the study. Compared with the control group, adding 0.5% dietary antimicrobial blend increased (P < 0.05) relative abundance of Firmicutes but reduced (P < 0.05) Bacteroidetes and Proteobacteria in feces on day 7 PI. Pigs that received the antimicrobial blend also had higher (P < 0.05) relative abundance of Lactobacillaceae, but lower (P < 0.05) relative abundance of Enterobacteriaceae in feces on day 7 PI than pigs in control. In conclusion, supplementation of this antimicrobial blend at 0.5% reduced incidence of severe diarrhea in weaned pigs challenged with F18 ETEC and enhanced feed efficiency of weaned pigs at the last week of the experiment. Supplementation of this antimicrobial blend also modified the microbiota diversity in feces and ileal mucosa of weaned pigs.

This experiment aims to investigate an antimicrobial blend consisting of essential oils, medium-chain fatty acids, and a toxin-adsorbing mineral on diarrhea, growth performance, and gut microbiome of newly weaned pigs experimentally infected with a pathogenic Escherichia coli (F18 E. coli). A total of 36 weaned pigs were randomly allotted to one of three dietary treatments: (1) a complex control diet that met the nutrient requirement of weaned pigs; (2) supplementing 0.25% of the antimicrobial blend; and (3) 0.50% of the antimicrobial blend. The experiment lasted 28 d with 7 d adaptation and 21 d after the first F18 E. coli inoculation. Results of this experiment demonstrate that supplementation of this antimicrobial blend enhanced disease resistance of weaned pigs, as indicated by reduced frequency of diarrhea during the entire experimental period. An improved feed efficiency was also observed in pigs supplemented with antimicrobial blend at the last week of the experiment. In addition, feces collected on day 7 post-E. coli inoculation contained relatively more Lactobacillaceae but less Enterobacteriaceae when pigs were supplemented with this antimicrobial blend. In conclusion, supplementation of antimicrobial blend could reduce diarrhea of E. coli-infected pigs and modify fecal microbiome of weaned pigs during the peak of E. coli infection.

A Retrospective Study of Double Eyelid Surgery for Sunken Upper Eyelid in Chinese.

OBJECTIVE: The aim of this study was to explore the effective technique of double eyelid surgery for mild, moderate, and severe sunken upper eyelid in Chinese. METHODS: According to the degree of the sunken upper eyelid, different procedures were performed. For mild sunken upper eyelid, the key technique was named eyelid volume redistribution (EVR)-1 technique, which was advancing or folding the pretarsal orbicularis oculi muscle flap toward up to the upper margin of the incision. For moderate sunken upper eyelid, the EVR-2 technique included EVR-1 technique and releasing orbital fat toward the upper margin of the tarsus. The other surgical steps were the same as conventional double eyelid surgery. For severe sunken upper eyelid, 2 steps were needed. The first step was eyelid volume augmentation (EVA). This technique injected fat into the retroorbicularis oculi fat layer. The second step was conventional double eyelid surgery which was performed 3 to 6 months later. RESULTS: One hundred sixty-seven patients with sunken upper eyelid underwent double eyelid surgery. In addition to the ameliorative recontouring of the sunken deformity, natural double eyelid was created. All patients had no complication, such as infection, skin irregularities, and lumps. The edema, bruising, and temporary ptosis recovered in 2 weeks. The edema resulted from double eyelid surgery, which last about 2 to 3 months. CONCLUSIONS: Our study showed that EVR-1, EVR-2, and EVA techniques have stable surgical results for mild, moderate, and severe sunken upper eyelids in Chinese. The procedures are simple, safe, and have less complications and preferable clinical reference value.

Determination of aucubin by supramolecular solvent-based dispersive liquid-liquid microextraction and UPLC-MS/MS: Application to a pharmacokinetic study in rats with type 1 diabetes.

A novel method was developed for the determination of aucubin in rat serum by ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) with supramolecular solvent (SUPRAS)-based dispersive liquid-liquid microextraction (DLLME). The SUPRAS was prepared with pentanol as extraction solvent and tetrahydrofuran as dispersing agent. Based on single factor test and response surface methodology, critical parameters were optimized as: 1 mL of pentanol as extraction solvent, 4 mL of tetrahydrofuran as dispersing agent, 200 µL of SUPRAS, and vortex duration of 2.5 min. The established method was validated in terms of selectivity, linearity, accuracy, precision, recovery, stability, and applied to a pharmacokinetic study on type 1 diabetes model rats intraperitoneally administered with aucubin. The experimental results showed that the maximum concentration in serum (Cmax) and area under the serum concentration versus time curve (AUC) of aucubin in type 1 diabetic rats were higher than those in normal rats. Such pharmacokinetic alterations might presumably result from the pathological state of type 1 diabetes. The current study may provide scientific evidence and inspiring insights for clinical application of aucubin for the treatment of diabetes.

Vertical enlargement of the palpebral aperture by surgical modification of the lower eyelid: A new cosmetic option for Chinese patients.

BACKGROUND: The distinctive features of Oriental eyes are narrow palpebral aperture and upslanting lower eyelid margin, which are not in accordance with modern appreciation of beauty. Although many ophthalmic plastic procedures have been designed to change the characteristic appearance, the methods to enlarge the palpebral aperture by lowering the lower eyelid are limited. METHODS: A total of 63 Chinese patients received the lowering the lower eyelid procedure from April 2014 to August 2018. The main criteria are patients who have vertically narrow palpebral aperture with or without upslanting lower eyelid shape. But patients who have proptosis or unhealthy lower eyelid elasticity are not suitable for this procedure. The operation is performed by suturing the lower tarsal plate and the infraorbital periosteum together, adjusting the tension of knots to reach the patient's desire for the shape of lower eyelid margin, and finally tightening all the knots. RESULTS: The lateral lower eyelid margin was lowered and the lateral part of the palpebral aperture was enlarged in all cases (P < 0.01). Only five patients (7.9%) were not fully satisfied because of the partial retraction of the lateral eyelid margin. Minor complications were observed, of which conjunctival chemosis in three patients (4.8%) and conjunctival hemorrhage in two patients (3.2%). CONCLUSION: Lowering the lower eyelid procedure is an effective and safe approach for patients who desire to smooth the upslanting shape and enlarge the eyes. The strict criteria and careful preoperative evaluations are critical to avoid complications and achieve good outcomes for Chinese patients.