Botulinum Toxin Type A Injection for Correction of Upper Eyelid Retraction in Thyroid-Associated Ophthalmopathy.

Upper lid retraction (ULR) is the most common and earliest symptom in thyroid-associated ophthalmopathy (TAO) patients. Surgical correction is effective for ULR in stable diseases. However, non-invasive treatment is also required for the TAO patient in active phase. Here, we reported a complex case with TAO and unilateral ULR simultaneously. The patient had a history of progressive ptosis in the left eyelid and underwent anterior levator aponeurotic-Muller muscle resection to correct the ptosis. However, the patient gradually developed bilateral proptosis and ULR, mainly in the left eyelid. The patient was finally diagnosed with TAO with left ULR. Then, the patient was treated with botulinum toxin type A (BTX-A) injection in the left eyelid. The effect of BTX-A treatment started 7 days after injection, peaked at one month, and lasted for approximately 3 months. This study highlighted the therapeutic effect of BTX-A injection for the treatment of ULR-related TAO.

Exopolysaccharide synthesis repressor genes (exoR and exoX) related to curdlan biosynthesis by Agrobacterium sp.

Curdlan is a neutral, water-insoluble, unbranched, linear ß-(1,3)-glucan. This study explored the roles of exoR and exoX in curdlan biosynthesis in Agrobacterium sp. ATCC 31749. The microcapsule biosynthesis of ΔexoR strain was reduced, and the motility of this strain increased remarkably compared with the wild-type (WT) strain during the cell growth phase. The curdlan yields of ΔexoR and ΔexoX strains enhanced by 19% and 17%, and the glucose utilization increased by 12% and 11%, respectively, compared with the WT strain during batch fermentation. By contrast, the curdlan yields of exoR and exoX overexpression strains decreased by 28% and 33%, respectively. The gel strength produced by ΔexoR and exoX overexpression strains decreased compared with the WT strain. RT-qPCR analysis at the transcriptional level revealed that key genes in exopolysaccharide synthesis and central metabolic pathways were up-regulated in ΔexoX and ΔexoR strains during gel production. Metabolomics analysis of ΔexoR and ΔexoX mutants proved the rates of central metabolic and electron transport chain were accelerated.

Preparation of recombinant Kluyveromyces lactis agents for simultaneous degradation of two mycotoxins.

Aflatoxin B1 (AFB1) and zearalenone (ZEN) are widely distributed in corns, peanuts, and other cereals, causing serious threat to food safety and human health. As shown by our previous studies, the recombinant yeast strain Kluyveromyces lactis GG799(pKLAC1-ZPF1) had the ability of degrading AFB1 and ZEN simultaneously. In this work, the agent preparation process was optimized for K. lactis GG799(pKLAC1-ZPF1), and the storage conditions of the prepared yeast agents were investigated, for obtaining the products with high storage activities and potent mycotoxin degradation efficiency. The optimal preparation process was as follows: centrifugation at 6000 rpm for 15 min for collection of the yeast cells, spray drying with the ratio of protective compounds to yeast cells at 3:1 (w/w) and then stored at - 20 °C. Simultaneous degradation tests of AFB1 and ZEN were performed using the supernatants of reactivated yeast agents after three months of storage, and the degradation ratios for AFB1 and ZEN in reaction system 1 (70.0 mmol/L malonic buffer, pH 4.5, with 1.0 mmol/L MnSO4, 0.1 mmol/L H2O2, 5.0 µg/mL AFB1 and ZEN, respectively) were 48.2 ± 3.2% and 34.8 ± 2.8%, while that for ZEN in reaction system 2 (50.0 mmol/L Tris-HCl, pH 7.5, with 5.0 µg/mL AFB1 and ZEN, respectively) was 30.1 ± 2.7%. Besides, the supernatants of reactivated yeast agents degraded more than 80% of AFB1 and 55% of ZEN in contaminated peanuts after twice treatments. Results of this work suggested that the optimized process for K. lactis GG799(pKLAC1-ZPF1) was with high value for industrial applications.

Efficient precious metal Rh(III) adsorption by waste P. pastoris and P. pastoris surface display from high-density culture.

Rhodium is one of the most used precious metals in catalysis both in laboratory reactions and industrial processes. However, the adsorption of Rh(III) by microorganisms has been seldomly reported. In this work, waste P. pastoris and recombinant P. pastoris with surface-displayed Rh metal peptides (P. pastoris GS-R) from high-density culture were used as novel adsorbents to study Rh(III) adsorption. Under the optimal adsorption conditions of biomass of 0.25 g L-1, pH of 1.2, temperature of 30 °C, and adsorption time of 2 h in simulated wastewater, the maximum adsorption ratios of 55.69% (110.1 mg g-1) and 75.03% (142.11 mg g-1) were achieved for waste P. pastoris and P. pastoris GS-R, respectively. Using the two adsorbents, the adsorption kinetic models fit the quasi-second-order equation, and isotherm models followed the Langmuir and Temkin equations, respectively. P. pastoris GS-R showed high adsorption capacity (48.49%) and selectivity (65.86%) in electroplating wastewater, and the desorption ratio reached 34.49% after treatment with 2 M HNO3 and ultrasonic wave. Therefore, an environmental-friendly strategy was developed for the recovery of Rh(III) and other precious metals by using P. pastoris.

Production of prebiotic gellan oligosaccharides based on the irradiation treatment and acid hydrolysis of gellan gum.

Biologically active gellan oligosaccharides (GOSs), newly found plant elicitors and biostimulants, are produced from the hydrolysis of gellan gum. Traditional hydrolysis with concentrated acid suffers from the problems of high pollution and low functional oligosaccharide yield because the process is difficult to control. Irradiation (60Co γ-ray) with a dosage ranging from 0 kGy to 175 kGy was used to degrade gellan gum efficiently and cleanly into low molecular weight (Mw) gellan with an average Mw ranging from 449,119 Da to 72,903 Da. The low Mw gellan irradiated at 70 kGy was further hydrolyzed with low concentration acid (0.5 mol/L HCl) to produce GOSs with DPs mainly 4 and 8, indicating that the Rha-ß-(1 â†’ 3)-Glc bonds in gellan gum were easily cut to produce residues with tetrasaccharide repeat subunits. Besides antioxidant activity, GOSs were also proved with prebiotic activity by in vitro fecal fermentation in a self-designed bionic intestinal reactor.

Environmental life cycle assessment of monosodium glutamate production in China: Based on the progress of cleaner production in recent ten years.

China is the world's leading producer of the flavor-enhancer monosodium glutamate (MSG), which is mainly produced by maize starch fermentation. Four typical scenarios (Scenario 1-Scenario 4) of MSG production from 2008 to 2019 in China were established for life cycle environmental impact assessment. Values for five midpoint environmental impact categories [primary energy demand (PED), resource depletion-water use (WU), global warming potential (GWP), acidification potential (AP), and eutrophication potential (EP)] were calculated with eFootprint based on data collected in China. The environmental impacts of producing 1 t of MSG in China in terms of PED, WU, GWP, AP, and EP were 52,800 MJ, 169,000 kg, 4590 kg CO2 eq, 42.1 kg SO2 eq, and 6.35 kg PO43- eq, respectively, in 2019. The life cycle of MSG from cradle to gate includes maize cultivation (high WU values), and the MSG production process (high PED, GWP, AP, and EP values). The normalization results of environmental impacts significantly decreased from S1 to S4 with the improvement in cleaner production in recent ten years. Sensitivity and uncertainty analyses identified steam for MSG production, irrigation water, and ammonia volatilization during maize growth as the three main factors affecting the environment. There were obvious differences in calculated environmental impacts depending on the maize cultivation data source; i.e., data from the Ecoinvent 3.1 database, or collected data for the maize-wheat rotation system in north China or the maize monoculture system in northeastern China. This demonstrates the importance of collecting local data rather than applying database from other world regions. Compared with the use of coal and natural gas, the use of biogas to produce steam had much lower PED, WU, and GWP. Which types of alternative energy to use and the replacement ratio should be based on the actual resource reserves in each region.

Research Advances of d-allulose: An Overview of Physiological Functions, Enzymatic Biotransformation Technologies, and Production Processes.

d-allulose has a significant application value as a sugar substitute, not only as a food ingredient and dietary supplement, but also with various physiological functions, such as improving insulin resistance, anti-obesity, and regulating glucolipid metabolism. Over the decades, the physiological functions of d-allulose and the corresponding mechanisms have been studied deeply, and this product has been applied to various foods to enhance food quality and prolong shelf life. In recent years, biotransformation technologies for the production of d-allulose using enzymatic approaches have gained more attention. However, there are few comprehensive reviews on this topic. This review focuses on the recent research advances of d-allulose, including (1) the physiological functions of d-allulose; (2) the major enzyme families used for the biotransformation of d-allulose and their microbial origins; (3) phylogenetic and structural characterization of d-allulose 3-epimerases, and the directed evolution methods for the enzymes; (4) heterologous expression of d-allulose ketose 3-epimerases and biotransformation techniques for d-allulose; and (5) production processes for biotransformation of d-allulose based on the characterized enzymes. Furthermore, the future trends on biosynthesis and applications of d-allulose in food and health industries are discussed and evaluated in this review.

One-step production of functional branched oligoglucosides with coupled fermentation of Pichia pastoris GS115 and Sclerotium rolfsii WSH-G01.

Endo-ß-1,3-glucanase with high specific activity is a prerequisite for enzymatic preparation of valuable ß-oligoglucosides. Heterologous expression in Pichia pastoris GS115 with error-prone PCR technology was implemented, and the mutant strain 7 N12 was obtained. The mutant endo-ß-1,3-glucanase showed efficient specific activities for degrading curdlan (366 U mg-1) and scleroglucan (274.5 U mg-1). Thereafter, one-step production of functional branched oligoglucosides was established with coupled fermentation of Pichia pastoris and Sclerotium rolfsii. During the fermentation process, the endo-ß-1,3-glucanase secreted by Pichia pastoris GS115 can efficiently hydrolyse scleroglucan metabolized by Sclerotium rolfsii WSH-G01. The maximum yields of ß-oligoglucosides in the shake flasks and 7-L bioreactor reached 1.73 g L-1 and 12.71 g L-1, respectively, with polymerization degrees of 2-17. The successful implementation of heterologous expression with error-prone PCR and the coupled fermentation simplified the multi-step enzymatic ß-oligoglucoside preparation procedures, which makes it a potential strategy for industrial production of functional oligosaccharides.

Efficient endo-ß-1,3-glucanase expression in Pichia pastoris for co-culture with Agrobacterium sp. for direct curdlan oligosaccharide production.

The production of curdlan oligosaccharides, a multifunctional and valuable carbohydrate, by hydrolyzing polysaccharides is of great interest. The endo-ß-1,3-glucanase derived from Trichoderma harzianum was expressed in Pichia pastoris with three commonly used promoters (AOX1, GAP and FLD1). The purified recombinant endo-ß-1,3-glucanase expressed by Pichia pastoris with GAP promoter displayed high specific activity at pH 5.5 and 50 °C. Thereafter, a co-culture system of Pichia pastoris GS115 (GAP promoter) and Agrobacterium sp. was constructed in which Agrobacterium sp.-metabolized curdlan can be directly hydrolyzed by Pichia pastoris-secreted endo-ß-1,3-glucanase to produce functional curdlan oligosaccharides. The co-culture conditions were optimized and the process was carried out in a 7-L bioreactor. The maximum yield of curdlan oligosaccharides reached 18.77 g/L with 3-10 degrees of polymerization. This study presents a novel and easy curdlan oligosaccharide production strategy that can replace traditional sophisticated production procedures and could potentially be implemented for production of other oligosaccharides.

High grade trophectoderm is associated with monozygotic twinning in frozen-thawed single blastocyst transfer.

BACKGROUND: The aim of this study was to explore specific factors that predispose to monozygotic twinning (MZT) at the blastocyst stage. METHODS: This was a retrospective observational study of a cohort of 2863 pregnancies after single blastocyst transfer (SBT) between January 2011 and June 2019 in our hospital. MZT pregnancy was identified as the number of fetuses exceeded the number of gestational sacs (GSs) by transvaginal ultrasound at 6-7 gestational weeks. The incidences of MZT regarding the maternal age at oocyte retrieval, paternal age, ovarian stimulation protocol, fertilization method, endometrium preparation protocol, vitrified day, and the Gardner grading of the blastocyst were calculated. The serum estrogen (E2), progesterone (P) levels, endometrium thickness and serum hCG levels on day 11 after embryo transfer (ET) were compared between the MZT and singleton pregnancies. Statistical analyses were used appropriately. RESULTS: Fifty-one MZT pregnancies (1.78%) were identified. The only significant differences observed between MZT and singleton pregnancies were the proportion of TE grade (P = 0.022) and the hCG levels on day 11 after ET (P = 0.003). Multivariate logistic regression revealed that trophectoderm (TE) grade was an independent factor affecting MZT, the adjusted odds ratios (aORs) of grade A and B TE were 5.46 [95% confidential interval (CI) 1.48-20.16, P = 0.011) and 3.96 (95% CI 1.17-13.40, P = 0.027) compared to grade C respectively. There were no significant associations between the parental age, fertilization method, ovarian stimulation protocol, endometrium preparation protocol, vitrified day, expansion stage, inner cell mass (ICM) grade and MZT. CONCLUSIONS: TE grade is associated with MZT at the blastocyst stage, potentially mediated via increased secretion of hCG from more well developed TE. Increased hCG secretion in turn may prolong the implantation window to support the embryo splitting.

Food-Grade Expression of Manganese Peroxidases in Recombinant Kluyveromyces lactis and Degradation of Aflatoxin B1 Using Fermentation Supernatants.

Aflatoxins are naturally occurring high-toxic secondary metabolites, which cause worldwide environmental contaminations and wastes of food and feed resources and severely threaten human health. Thus, the highly efficient methods and technologies for detoxification of aflatoxins are urgently needed in a long term. In this work, we report the construction of recombinant Kluyveromyces lactis strains GG799(pKLAC1-Phsmnp), GG799(pKLAC1-Plomnp), GG799(pKLAC1-Phcmnp), and then the food-grade expression of the three manganese peroxidases in these strains, followed by the degradation of aflatoxin B1 (AFB1) using the fermentation supernatants. The expression of the manganese peroxidases was achieved in a food-grade manner since Kluyveromyces lactis is food-safe and suitable for application in food or feed industries. The inducible expression process of the optimal recombinant strain GG799(pKLAC1-Phcmnp) and the aflatoxin B1 degradation process were both optimized in detail. After optimization, the degradation ratio reached 75.71%, which was an increase of 49.86% compared to the unoptimized results. The degradation product was analyzed and determined to be AFB1-8,9-dihydrodiol. The recombinant strain GG799(pKLAC1-Phcmnp) supernatants degraded more than 90% of AFB1 in the peanut samples after twice treatments. The structural computational analysis for further mutagenesis of the enzyme PhcMnp was also conducted in this work. The food-grade recombinant yeast strain and the enzyme PhcMnp have potential to be applied in food or feed industries.

Pregnancy Outcomes of Women With Polycystic Ovary Syndrome for the First In Vitro Fertilization Treatment: A Retrospective Cohort Study With 7678 Patients.

Background: The risk of adverse pregnancy outcomes is increased by having a polycystic ovary syndrome (PCOS) diagnosis. However, the confounders in previous studies preclude firm conclusions, and further studies are warranted. Objectives: To investigate whether PCOS affects pregnancy outcomes and complications in infertile women undergoing their first in vitro fertilization (IVF) treatment, taking into account important confounders. Methods: We performed a retrospective cohort study of 7,678 infertile women, including 666 women with PCOS and 7,012 controls undergoing their first IVF treatment at a private fertility center from January 2010 to December 2017. Our main outcome was the impact of PCOS on adverse pregnancy outcomes (miscarriage, preterm delivery, pregnancy-induced hypertension) and pregnancy outcomes (live birth rate, clinical pregnancy rate, implantation rate). PCOS effects were summarized by adjusted odds ratios (aORs) with 95% confidence intervals (CIs) after controlling for maternal characteristics. Results: After adjusting for differences in maternal age, BMI, infertility duration, total dose of gonadotropin, serum E2 and endometrial thickness on the day of hCG trigger, number of fertilized occytes, number of embryos transferred, embryo type (cleavage-stage embryo or blastocyst) and quality, women with PCOS had an increased risk of developing unfavorable pregnancy complications, including miscarriage (aOR 1.629, 95% CI 1.240-2.141), very preterm delivery (< 32 weeks) (aOR 2.072, 95% CI 1.133-3.791). For pregnancy outcomes, PCOS was associated with higher clinical pregnancy rate (aOR 1.248, 95% CI 1.038-1.501) and implantation rate (aOR 1.238, 95% CI 1.030-1.489) after adjusting for the above-mentioned confounders. Conclusions: Women with PCOS are at increased risk of adverse pregnancy outcomes after adjusting for differences in maternal characteristics. These women may need more frequent medical consultants and management during pregnancy and parturition.

Maternal chronic hepatitis B virus infection does not affect pregnancy outcomes in infertile patients receiving first in vitro fertilization treatment.

OBJECTIVE: To evaluate whether maternal chronic hepatitis B virus (HBV) infection affects pregnancy outcomes in infertile patients undergoing their first in vitro fertilization (IVF) treatment. DESIGN: A retrospective case control study. SETTING: Fertility center. PATIENT(S): Female patients, comprising 8,550 infertile women including 180 HBsAg+HBeAg+, 714 HBsAg+HBeAg-, and 7,656 HBsAg seronegative controls undergoing their first IVF treatments. INTERVENTION(S): Clinical characteristics, pregnancy and neonatal outcomes were analyzed by Kruskal-Wallis test, analysis of variance, or chi-square test. Logistic regression was employed to verify the contribution of maternal HBV to clinical pregnancy, live birth, and miscarriage. MAIN OUTCOME MEASURE(S): Primary outcome: live-birth rate; secondary outcomes: implantation, clinical pregnancy, and miscarriage rates. RESULT(S): An increased duration of infertility and more secondary infertility and ovulatory disorders were observed in the HBV patients. The implantation rate was statistically significantly lower in the HBsAg+HBeAg- group compared with the controls. However, the clinical pregnancy rate, miscarriage rate, live-birth rate, neonatal outcomes, and pregnancy complications showed no statistically significant differences among the groups. The logistic regression analysis showed that HBV infection status did not affect the clinical pregnancy, miscarriage, or live-birth rates, unlike maternal age, endometrial thickness, and use of high-quality embryos. CONCLUSION(S): Hepatitis B virus infection is not an independent contributor to pregnancy outcomes, although it is associated with prolonged infertility duration, a high frequency of secondary infertility and ovulatory disorders, and a reduced implantation rate.

The Influence of Different Knowledge-Driven Methods on Landslide Susceptibility Mapping: A Case Study in the Changbai Mountain Area, Northeast China.

Landslides are one of the most frequent geomorphic hazards, and they often result in the loss of property and human life in the Changbai Mountain area (CMA), Northeast China. The objective of this study was to produce and compare landslide susceptibility maps for the CMA using an information content model (ICM) with three knowledge-driven methods (the artificial hierarchy process with the ICM (AHP-ICM), the entropy weight method with the ICM (EWM-ICM), and the rough set with the ICM (RS-ICM)) and to explore the influence of different knowledge-driven methods for a series of parameters on the accuracy of landslide susceptibility mapping (LSM). In this research, the landslide inventory data (145 landslides) were randomly divided into a training dataset: 70% (81 landslides) were used for training the models and 30% (35 landslides) were used for validation. In addition, 13 layers of landslide conditioning factors, namely, altitude, slope gradient, slope aspect, lithology, distance to faults, distance to roads, distance to rivers, annual precipitation, land type, normalized difference vegetation index (NDVI), topographic wetness index (TWI), plan curvature, and profile curvature, were taken as independent, causal predictors. Landslide susceptibility maps were developed using the ICM, RS-ICM, AHP-ICM, and EWM-ICM, in which weights were assigned to every conditioning factor. The resultant susceptibility was validated using the area under the ROC curve (AUC) method. The success accuracies of the landslide susceptibility maps produced by the ICM, RS-ICM, AHP-ICM, and EWM-ICM methods were 0.931, 0.939, 0.912, and 0.883, respectively, with prediction accuracy rates of 0.926, 0.927, 0.917, and 0.878 for the ICM, RS-ICM, AHP-ICM, and EWM-ICM, respectively. Hence, it can be concluded that the four models used in this study gave close results, with the RS-ICM exhibiting the best performance in landslide susceptibility mapping.

A Method for Improving Controlling Factors Based on Information Fusion for Debris Flow Susceptibility Mapping: A Case Study in Jilin Province, China.

Debris flow is one of the most frequently occurring geological disasters in Jilin province, China, and such disasters often result in the loss of human life and property. The objective of this study is to propose and verify an information fusion (IF) method in order to improve the factors controlling debris flow as well as the accuracy of the debris flow susceptibility map. Nine layers of factors controlling debris flow (i.e., topography, elevation, annual precipitation, distance to water system, slope angle, slope aspect, population density, lithology and vegetation coverage) were taken as the predictors. The controlling factors were improved by using the IF method. Based on the original controlling factors and the improved controlling factors, debris flow susceptibility maps were developed while using the statistical index (SI) model, the analytic hierarchy process (AHP) model, the random forest (RF) model, and their four integrated models. The results were compared using receiver operating characteristic (ROC) curve, and the spatial consistency of the debris flow susceptibility maps was analyzed while using Spearman's rank correlation coefficients. The results show that the IF method that was used to improve the controlling factors can effectively enhance the performance of the debris flow susceptibility maps, with the IF-SI-RF model exhibiting the best performance in terms of debris flow susceptibility mapping.