Enhanced Enzymatic Performance of Immobilized Pseudomonas fluorescens Lipase on ZIF-8@ZIF-67 and Its Application to the Synthesis of Neryl Acetate with Transesterification Reaction.

In this study, hybrid skeleton material ZIF-8@ZIF-67 was synthesized by the epitaxial growth method and then was utilized as a carrier for encapsulating Pseudomonas fluorescens lipase (PFL) through the co-precipitation method, resulting in the preparation of immobilized lipase (PFL@ZIF-8@ZIF-67). Subsequently, it was further treated with glutaraldehyde to improve protein immobilization yield. Under optimal immobilization conditions, the specific hydrolytic activity of PFL@ZIF-8@ZIF-67 was 20.4 times higher than that of the free PFL. The prepared biocatalyst was characterized and analyzed by scanning electron microscopy (SEM), X-ray diffraction (XRD), and Fourier transform infrared (FT-IR). Additionally, the thermal stability of PFL@ZIF-8@ZIF-67 at 50 °C was significantly improved compared to the free PFL. After 7 weeks at room temperature, PFL@ZIF-8@ZIF-67 retained 78% of the transesterification activity, while the free enzyme was only 29%. Finally, PFL@ZIF-8@ZIF-67 was applied to the neryl acetate preparation in a solvent-free system, and the yield of neryl acetate reached 99% after 3 h of reaction. After 10 repetitions, the yields of neryl acetate catalyzed by PFL@ZIF-8@ZIF-67 and the free PFL were 80% and 43%, respectively.

Histone H3K18 and Ezrin Lactylation Promote Renal Dysfunction in Sepsis-Associated Acute Kidney Injury.

Histone lactylation is a metabolic stress-related histone modification. However, the role of histone lactylation in the development of sepsis-associated acute kidney injury (SA-AKI) remains unclear. Here, histone H3K18 lactylation (H3K18la) is elevated in SA-AKI, which is reported in this study. Furthermore, this lactate-dependent histone modification is enriched at the promoter of Ras homolog gene family member A (RhoA) and positively correlated with the transcription. Correction of abnormal lactate levels resulted in a reversal of abnormal histone lactylation at the promoter of RhoA. Examination of related mechanism revealed that histone lactylation promoted the RhoA/Rho-associated protein kinase (ROCK) /Ezrin signaling, the activation of nuclear factor-κB (NF-κB), inflammation, cell apoptosis, and aggravated renal dysfunction. In addition, Ezrin can undergo lactylation modification. Multiple lactylation sites are identified in Ezrin and confirmed that lactylation mainly occurred at the K263 site. The role of histone lactylation is revealed in SA-AKI and reportes a novel post-translational modification in Ezrin. Its potential role in regulating inflammatory metabolic adaptation of renal proximal tubule epithelial cells is also elucidated. The results provide novel insights into the epigenetic regulation of the onset of SA-AKI.

A fully automatic fiducial detection and correspondence establishing method for online C-arm calibration.

PURPOSE: Online C-arm calibration with a mobile fiducial cage plays an essential role in various image-guided interventions. However, it is challenging to develop a fully automatic approach, which requires not only an accurate detection of fiducial projections but also a robust 2D-3D correspondence establishment. METHODS: We propose a novel approach for online C-arm calibration with a mobile fiducial cage. Specifically, a novel mobile calibration cage embedded with 16 fiducials is designed, where the fiducials are arranged to form 4 line patterns with different cross-ratios. Then, an auto-context-based detection network (ADNet) is proposed to perform an accurate and robust detection of 2D projections of those fiducials in acquired C-arm images. Subsequently, we present a cross-ratio consistency-based 2D-3D correspondence establishing method to automatically match the detected 2D fiducial projections with those 3D fiducials, allowing for an accurate online C-arm calibration. RESULTS: We designed and conducted comprehensive experiments to evaluate the proposed approach. For automatic detection of 2D fiducial projections, the proposed ADNet achieved a mean point-to-point distance of 0.65 ± 1.33 pixels. Additionally, the proposed C-arm calibration approach achieved a mean re-projection error of 1.01 ± 0.63 pixels and a mean point-to-line distance of 0.22 ± 0.12  mm. When the proposed C-arm calibration approach was applied to downstream tasks involving landmark and surface model reconstruction, sub-millimeter accuracy was achieved. CONCLUSION: In summary, we developed a novel approach for online C-arm calibration. Both qualitative and quantitative results of comprehensive experiments demonstrated the accuracy and robustness of the proposed approach. Our approach holds potentials for various image-guided interventions.

Multiple phenotype association tests based on sliced inverse regression.

BACKGROUND: Joint analysis of multiple phenotypes in studies of biological systems such as Genome-Wide Association Studies is critical to revealing the functional interactions between various traits and genetic variants, but growth of data in dimensionality has become a very challenging problem in the widespread use of joint analysis. To handle the excessiveness of variables, we consider the sliced inverse regression (SIR) method. Specifically, we propose a novel SIR-based association test that is robust and powerful in testing the association between multiple predictors and multiple outcomes. RESULTS: We conduct simulation studies in both low- and high-dimensional settings with various numbers of Single-Nucleotide Polymorphisms and consider the correlation structure of traits. Simulation results show that the proposed method outperforms the existing methods. We also successfully apply our method to the genetic association study of ADNI dataset. Both the simulation studies and real data analysis show that the SIR-based association test is valid and achieves a higher efficiency compared with its competitors. CONCLUSION: Several scenarios with low- and high-dimensional responses and genotypes are considered in this paper. Our SIR-based method controls the estimated type I error at the pre-specified level α .

Circ DENND4C inhibits pyroptosis and alleviates ischemia-reperfusion acute kidney injury by exosomes secreted from human urine-derived stem cells.

Acute kidney injury (AKI) is a disease characterised by acute onset, high mortality, and poor prognosis, and is mainly caused by ischemia-reperfusion (I/R). Human urine-derived stem cells (USCs) exhibit antioxidant, anti-inflammatory, and anti-apoptotic cytoprotective effects. Previously, we found that exosomes from USCs had the ability to inhibit apoptosis and protect kidneys from I/R injury. This study aimed to investigate the role of USC-derived exosomes (USC-Exos) in reducing pyroptosis and alleviating I/R-AKI. Models of HK-2 cells hypoxia-reoxygenation (H/R) and I/R kidney injury was established in Sprague Dawley rats to simulate AKI in vitro and in vivo. USC-Exos were isolated using ultracentrifugation and identified via electron microscopy and western blotting. USC-Exos were co-cultured with HK-2 cells and injected into rats via the tail vein. The expression of pyroptosis-related molecules (GSDMD, caspase-1, and NLRP-3) was verified using PCR and western blotting. Changes in renal function were reflected in the serum creatinine, urea, and cystatin C levels. The degree of renal injury was determined using haematoxylin and eosin and immunohistochemical staining. The levels of IL-1ß and IL-18 were detected using enzyme-linked immunosorbent assay (ELISA) to verify the role of USC-Exos in pyroptosis. Differentially expressed circRNAs in I/R rat kidneys were screened by transcriptome sequencing, and a dual-luciferase experiment was used to verify the interaction between upstream and downstream molecules. Ischemia-reperfusion resulted in significantly impaired renal function and expression of pyroptosis molecules, and significantly increased concentrations of inflammatory factors. These effects were reversed by injecting USC-Exos. Circ DENND4C was the most significantly decreased circRNA in I/R rat renal tissue, and knock-down of circ DENND4C can aggravate AKI in vivo and in vitro. DAVID(http://david.abcc.ncifcrf.gov) website showed that miR 138-5p/FOXO3a is a potential downstream target of circ DENND4C. Knock-down of circ DENND4C in HK-2 cells resulted in increased expression of miR 138-5p and increased miR 138-5p can reverse the regulation of FOXO3a. Dual-luciferase assay verified the reverse interaction between circ DENND4C, miR 138-5p, and FOXO3a. Exosomes promote cell proliferation and inhibit the activation of NLR family pyrin domain containing 3 through the circ DENND4C/miR 138-5p/FOXO3a pathway, thereby reducing pyroptosis and AKI. Circ DENND4C may be a potential therapeutic target for AKI.

Contextual Patch-NetVLAD: Context-Aware Patch Feature Descriptor and Patch Matching Mechanism for Visual Place Recognition.

The goal of visual place recognition (VPR) is to determine the location of a query image by identifying its place in a collection of image databases. Visual sensor technologies are crucial for visual place recognition as they allow for precise identification and location of query images within a database. Global descriptor-based VPR methods face the challenge of accurately capturing the local specific regions within a scene; consequently, it leads to an increasing probability of confusion during localization in such scenarios. To tackle feature extraction and feature matching challenges in VPR, we propose a modified patch-NetVLAD strategy that includes two new modules: a context-aware patch descriptor and a context-aware patch matching mechanism. Firstly, we propose a context-driven patch feature descriptor to overcome the limitations of global and local descriptors in visual place recognition. This descriptor aggregates features from each patch's surrounding neighborhood. Secondly, we introduce a context-driven feature matching mechanism that utilizes cluster and saliency context-driven weighting rules to assign higher weights to patches that are less similar to densely populated or locally similar regions for improved localization performance. We further incorporate both of these modules into the patch-NetVLAD framework, resulting in a new approach called contextual patch-NetVLAD. Experimental results are provided to show that our proposed approach outperforms other state-of-the-art methods to achieve a Recall@10 score of 99.82 on Pittsburgh30k, 99.82 on FMDataset, and 97.68 on our benchmark dataset.

Heterogeneous environmental policies and employment: evidence from China.

This paper attempts to investigate how the rapid development of China's environmental policy contributes to the changes in China's labour market. We first categorise the environmental regulations into (i) command-and-control policies, (ii) economic incentive policies, and (iii) voluntary environmental agreements (VEAs). Then, we use the provincial panel data over the 2000-2015 period to estimate the effect of alternative environmental regulations on employment in China. The results show that alternative environmental regulations have different impacts on employment. This difference even exists within the same group of environmental policies. Amongst the command-and-control policies, the three simultaneities policy leads to less employment, while the deadline governance systems increase employment. For economic incentive policies, higher environmental tax leads to lower employment, while more funding for environmental research promotes employment. The results also show that more VEAs reduced labour demand. In addition, the dynamic impact and regional heterogeneity of environmental regulations on employment are confirmed by the empirical estimation. Specifically, the economic incentive policies significantly impact employment in Central and Western China, whereas they have a non-significant impact on employment in Eastern China. This finding confirms that economic development impacts the relationship between environmental regulations and employment.

Serum exosomal microRNA-1258 may as a novel biomarker for the diagnosis of acute exacerbations of chronic obstructive pulmonary disease.

Acute exacerbation chronic obstructive pulmonary disease (AECOPD) has a high mortality rate. However, there is no efficiency biomarker for diagnosing AECOPD. The purpose of this study was to find biomarkers that can quickly and accurately diagnose AECOPD.45 normal controls (NC), 42 patients with stable COPD (SCOPD), and 66 patients with AECOPD were enrolled in our study. Serum exosomes were isolated by ultracentrifuge and verified by morphology and specific biomarkers. Fluorescent quantitation polymerase chain reaction (qRT-PCR) was used to detect the expression of micro RNAs (miRNAs), including miR-660-5p, miR-1258, miR-182-3p, miR-148a-3p, miR-27a-5p and miR-497-5p in serum exosomes and serum. Logistic regression and machine learning methods were used to constructed the diagnostic models of AECOPD. The levels of miR-1258 in the patients with AECOPD were higher than other groups (p < 0.001). The ability of exosomal miR-1258 (AUC = 0.851) to identify AECOPD from SCOPD was superior to other biomarkers, and the combination of exosomal miR-1258 and NLR can increase the AUC to 0.944, with a sensitivity of 81.82%, and specificity of 97.62%. The cross-validation of the models displayed that the logistic regression model based on exosomal miR-1258, NLR and neutrophil count had the best accuracy (0.880) in diagnosing AECOPD from SCOPD. The three most correlated biomarkers with serum exosome miR-1258 were neutrophil count (r = 0.57, p < 0.001), WBC (r = 0.50, p < 0.001) and serum miR-1258 (r = 0.33, p < 0.001). In conclusion, serum exosomal miR-1258 is associated with inflammation, and can be used as a valuable and reliable biomarker for the diagnosis of AECOPD, and the establishment of diagnostic model based on miR-1258, NLR and neutrophils count can help to improving the accuracy of AECOPD diagnosis.

Efficacy and safety of duloxetine in chronic musculoskeletal pain: a systematic review and meta-analysis.

BACKGROUND: Chronic musculoskeletal pain (CMP) is a complex condition that is mainly treated with analgesic drugs. However, antidepressant intervention is also an important factor in the treatment of CMP. Duloxetine is an effective treatment option for patients with CMP as its antidepressant effect. The purpose of this article is to evaluate the efficacy and safety of duloxetine in treating CMP. DATABASES AND DATA TREATMENT: We searched PubMed, Web of Science, Embase, Cochrane Library from inception to May, 2022. Randomized controlled trials (RCTs) evaluating the efficacy and safety of duloxetine versus placebo in patients with CMP were included. We identified 13 articles and studied a population of 4201 participants in 4 countries. RESULTS: This meta-analysis showed that the duloxetine has statistically significant compared with the placebo control, benefits on 24-hour average pain, living quality, physical function, and global impressions and there was no difference in the incidence of serious adverse event. In general, duloxetine can improve mood and pain level at the same time. CONCLUSIONS: This review shows a significant contribution of duloxetine to CMP symptom relief. This meta-analysis improved that duloxetine can significantly reduce the pain level of patients, improve depressive symptoms and global impression, and has no obvious serious adverse reactions. However, additional studies are required to confirm the relationship between psychological diseases and chronic pain and explore their internal links.

Improved Synthesis of Deoxyadenosine Triphosphate by Saccharomyces cerevisiae Using an Efficient ATP Regeneration System: Optimization of Response Surface Analysis.

Deoxyadenosine triphosphate (dATP) is an important biochemical molecule. In this paper, the synthesis of dATP from deoxyadenosine monophosphate (dAMP), catalyzed by Saccharomyces cerevisiae, was studied. By adding chemical effectors, an efficient ATP regeneration and coupling system was constructed to achieve efficient synthesis of dATP. Factorial and response surface designs were used to optimize process conditions. Optimal reaction conditions were as follows: dAMP 1.40 g/L, glucose 40.97 g/L, MgCl2·6H2O 4.00 g/L, KCl 2.00 g/L, NaH2PO4 31.20 g/L, yeast 300.00 g/L, ammonium chloride 0.67 g/L, acetaldehyde 11.64 mL/L, pH 7.0, temperature 29.6 °C. Under these conditions, the substrate conversion was 93.80% and the concentration of dATP in the reaction system was 2.10 g/L, which was 63.10% higher than before optimization, and the concentration of product was 4 times higher than before optimization. The effects of glucose, acetaldehyde, and temperature on the accumulation of dATP were analyzed.

LatentPCN: latent space-constrained point cloud network for reconstruction of 3D patient-specific bone surface models from calibrated biplanar X-ray images.

PURPOSE: Accurate three-dimensional (3D) models play crucial roles in computer assisted planning and interventions. MR or CT images are frequently used to derive 3D models but have the disadvantages that they are expensive or involving ionizing radiation (e.g., CT acquisition). An alternative method based on calibrated 2D biplanar X-ray images is highly desired. METHODS: A point cloud network, referred as LatentPCN, is developed for reconstruction of 3D surface models from calibrated biplanar X-ray images. LatentPCN consists of three components: an encoder, a predictor, and a decoder. During training, a latent space is learned to represent shape features. After training, LatentPCN maps sparse silhouettes generated from 2D images to a latent representation, which is taken as the input to the decoder to derive a 3D bone surface model. Additionally, LatentPCN allows for estimation of a patient-specific reconstruction uncertainty. RESULTS: We designed and conducted comprehensive experiments on datasets of 25 simulated cases and 10 cadaveric cases to evaluate the performance of LatentLCN. On these two datasets, the mean reconstruction errors achieved by LatentLCN were 0.83 mm and 0.92 mm, respectively. A correlation between large reconstruction errors and high uncertainty in the reconstruction results was observed. CONCLUSION: LatentPCN can reconstruct patient-specific 3D surface models from calibrated 2D biplanar X-ray images with high accuracy and uncertainty estimation. The sub-millimeter reconstruction accuracy on cadaveric cases demonstrates its potential for surgical navigation applications.

Ultrasound Probe and Hand-Eye Calibrations for Robot-Assisted Needle Biopsy.

In robot-assisted ultrasound-guided needle biopsy, it is essential to conduct calibration of the ultrasound probe and to perform hand-eye calibration of the robot in order to establish a link between intra-operatively acquired ultrasound images and robot-assisted needle insertion. Based on a high-precision optical tracking system, novel methods for ultrasound probe and robot hand-eye calibration are proposed. Specifically, we first fix optically trackable markers to the ultrasound probe and to the robot, respectively. We then design a five-wire phantom to calibrate the ultrasound probe. Finally, an effective method taking advantage of steady movement of the robot but without an additional calibration frame or the need to solve the AX=XB equation is proposed for hand-eye calibration. After calibrations, our system allows for in situ definition of target lesions and aiming trajectories from intra-operatively acquired ultrasound images in order to align the robot for precise needle biopsy. Comprehensive experiments were conducted to evaluate accuracy of different components of our system as well as the overall system accuracy. Experiment results demonstrated the efficacy of the proposed methods.

Evaluating China's pilot carbon Emission Trading Scheme: collaborative reduction of carbon and air pollutants.

Collaborative reduction of carbon and air pollutants can more efficiently achieve green technological change, industrial low-carbon transition, and high-quality economic and social development. As a typical environmental policy in China, the pilot carbon Emission Trading Scheme (ETS) has obvious advantages in achieving the collaborative reduction of carbon and air pollutants. Therefore, an evaluation of China's pilot carbon ETS from the perspective of collaborative reduction of carbon and air pollutants is performed in this paper. Compared with previous studies, first, this study innovatively uses the coupled coordination degree (CCD) model to measure the collaborative reduction level of carbon and air pollutants under different scenarios based on the panel data of China's 30 provincial-level regions during 2004-2018. Second, this study uses the DID method to evaluate the impact of China's pilot carbon ETS on the collaborative reduction of carbon and air pollutants and conducts some robustness checks and regional heterogeneity regressions. Third, this study uses the synthetic control method (SCM) further to examine the policy outcomes of the pilot carbon ETS. Scenario analysis shows that attaching importance to reducing air pollution will improve the collaborative reduction effect of carbon and air pollutants. Furthermore, the implementation of China's pilot carbon ETS exerts an effect of roughly 24.7% on reducing carbon, roughly 10.1% on reducing air pollutants, and roughly 22.0% on the collaborative reduction of carbon and air pollutants, ceteris paribus. Regional heterogeneity analysis shows that the impacts of the pilot carbon ETS are significant in all regions, except that the impact on reducing air pollutants in the central region is not significant. In addition, results from SCM indicate that the impacts of the pilot carbon ETS on the collaborative reduction of carbon and air pollutants are significantly efficient in Beijing, Tianjin, Shanghai, Hubei, and Chongqing, while not much efficient in Guangdong and Fujian. The main policy implications include strengthening the top-level design of the ETS in the collaborative reduction of carbon and air pollutants, attaching importance to the governance of air pollution, making the regional governance more targeted, and improving energy efficiency.

A Novel Point Set Registration-Based Hand-Eye Calibration Method for Robot-Assisted Surgery.

Pedicle screw insertion with robot assistance dramatically improves surgical accuracy and safety when compared with manual implantation. In developing such a system, hand-eye calibration is an essential component that aims to determine the transformation between a position tracking and robot-arm systems. In this paper, we propose an effective hand-eye calibration method, namely registration-based hand-eye calibration (RHC), which estimates the calibration transformation via point set registration without the need to solve the AX=XB equation. Our hand-eye calibration method consists of tool-tip pivot calibrations in two-coordinate systems, in addition to paired-point matching, where the point pairs are generated via the steady movement of the robot arm in space. After calibration, our system allows for robot-assisted, image-guided pedicle screw insertion. Comprehensive experiments are conducted to verify the efficacy of the proposed hand-eye calibration method. A mean distance deviation of 0.70 mm and a mean angular deviation of 0.68° are achieved by our system when the proposed hand-eye calibration method is used. Further experiments on drilling trajectories are conducted on plastic vertebrae as well as pig vertebrae. A mean distance deviation of 1.01 mm and a mean angular deviation of 1.11° are observed when the drilled trajectories are compared with the planned trajectories on the pig vertebrae.

Analytical and clinical performance evaluation of a new high-sensitivity cardiac troponin I assay.

OBJECTIVES: To validate the analytical performance and diagnostic accuracy for non-ST-segment elevation myocardial infarction (NSTEMI) with a new high-sensitivity cardiac troponin I (hs-cTnI) assay on the automated light-initiated chemiluminescent assay (LiCA®) platform. METHODS: Comprehensive analytical validations were performed, and the 99th percentile upper reference limit (URL) from apparently healthy individuals were established. We evaluated the diagnostic performance of the assay for NSTEMI. RESULTS: The limit of quantitation (LoQ) were 1.9 ng/L (20% CV) and 5.1 ng/L (10% CV). The sex-specific 99th percentile URLs were 17.6 ng/L (4.2% CV) for men (age 20-79y) and 14.2 ng/L (4.9% CV) for women (age 19-89y) in serum, 14.4 ng/L (4.9% CV) for men (age 19-88y) and 12.9 ng/L (5.2% CV) for women (age 19-87y) in plasma, respectively. Detection rates in healthy individuals were from 98.7 to 99.1%. The correlation coefficient and median bias between LiCA and Architect were 0.985 and 0.1% (-2.0-2.9%) in full analytical range of serum specimens. In lower range (<100 ng/L), LiCA had an overall positive bias 6.7% (-1.6-13.3%), R=0.949. At the specific medical decision levels (15.2, 26.2 and 64.0 ng/L), assay difference was estimated to be <10%. No significant differences on AUC, sensitivity and specificity, NPV and PPV were found between LiCA and Architect for the diagnosis of NSTEMI. CONCLUSIONS: LiCA hs-cTnI is a precise, highly sensitive and specific assay that meets the requirement of a 3rd generation (level 4) high-sensitivity method. The diagnostic accuracy of LiCA assay for NSTEMI is comparable to the established Architect hs-cTnI assay.

An Ultrafast, Durable, and High-Loading Polymer Anode for Aqueous Zinc-Ion Batteries and Supercapacitors.

Zn metal has shown promise as an anode material for grid-level energy storage, yet is challenged by dendritic growth and low Coulombic efficiency. Herein, an ultrafast, stable, and high-loading polymer anode for aqueous Zn-ion batteries and capacitors (ZIBs and ZICs) is developed by engineering both the electrode and electrolyte. The anode polymer is rationally prepared to have a suitable electronic structure and a large π-conjugated structure, whereas the electrolyte is manufactured based on the superiority of triflate anions over sulfate anions, as analyzed and confirmed via experiments and simulations. This dual engineering results in an optimal polymer anode with a low discharge potential, near-theoretical capacity, ultrahigh-loading capability (≈50 mg cm-2 ), ultrafast rate (100 A g-1 ), and ultralong lifespan (one million cycles). Its mechanism involves reversible Zn2+ /proton co-storage at the carbonyl site. When the polymer anode is coupled with cathodes for both ZIB and ZIC applications, the devices demonstrate ultrahigh power densities and ultralong lifespans, far surpassing those of corresponding Zn-metal-based devices.

Enhanced Adsorption of Anionic Polymer on Montmorillonite by Divalent Cations and the Effect of Salinity.

Adsorption of polymers from an aqueous solution onto clay minerals is of great interest to many applications such as water purification and soil conditioning. Molecular dynamics simulations were performed to study the adsorption of anionic polyacrylamide (APAM) on anionic montmorillonite, in an aqueous solution containing monovalent or divalent salts. Compared with monovalent salts (NaCl), the enhancement of APAM adsorption brought by divalent salts (CaCl2) was significant, which could not be explained by the Poisson-Boltzmann theory alone. Each solvated Ca2+ was coordinated by 4-6 water oxygens in its first coordination shell. One to two of these water molecules were displaced when APAM formed a complex with Ca2+. Ca2+ ions in the adsorbed Ca2+-APAM complexes did not serve as bridges sandwiched between APAM and Mt; instead, the complexes carried a residual positive charge and were subsequently attracted to montmorillonite. The number of adsorbed Ca2+-APAM complexes changed with salinity in a nonmonotonic manner, due to the modulation of apparent charges of montmorillonite and APAM by Ca2+. Increasing adsorption of Ca2+-APAM complexes also promoted APAM adsorption through direct hydrogen bonding with montmorillonite. The findings provided new molecular insights into the long-standing debates on the role of divalent ions in promoting polymer adsorption on like-charged solid surfaces.

The impact of environmental regulation on employment: an empirical study of China's Two Control Zone policy.

Environmental regulations affect employment through productivity output and factor substitution. This paper employs a difference-in-differences (DID) method to investigate the effect of China's Two Control Zones (TCZ) policy on the urban employment in 287 cities from 1994 to 2009. We apply the DID method to two time points: 1998 for policy issuance and 2000 for the policy implementation. From the results of analyses on full-sample cities, the TCZ policy did not contribute to increasing total urban employment. Moreover, a negative impact on employment resulted from sulfur dioxide and acid rain controls in secondary and tertiary industries, respectively. In the acid rain control zone, the TCZ policy increased the average wage of urban workers. Negative effects on employment were observed in larger cities. The policy triggered labor migration from larger to smaller cities, resulting in significant increases in primary and tertiary industry employment in smaller cities, although the effects on mid-size cities were insignificant. This study provides important empirical evidence and insight into the impact of the TCZ policy on urban employment.

Galactomannan Degrading Enzymes from the Mannan Utilization Gene Cluster of Alkaliphilic Bacillus sp. N16-5 and Their Synergy on Galactomannan Degradation.

Two glycoside hydrolases encoded by the mannan utilization gene cluster of alkaliphilic Bacillus sp. N16-5 were studied. The recombinant Gal27A (rGal27A) hydrolyzed both galactomannans and oligo-galactomannans to release galactose, while the recombinant Man113A (rMan113A) showed poor activity toward galactomannans, but it hydrolyzed manno-oligosaccharides to release mannose and mannobiose. rGal27A showed synergistic interactions with rMan113A and recombinant ß-mannanase ManA (rManA), which is also from Bacillus sp. N16-5, in galactomannan degradation. The synergy degree of rGal27A and rManA on hydrolysis of locust bean gum and guar gum was 1.13 and 2.21, respectively, and that of rGal27A and rMan113A reached 2.00 and 2.68. The main products of galactomannan hydrolyzed by rGal27A and rManA simultaneously were galactose, mannose, mannobiose, and mannotriose, while those of galactomannan hydrolyzed by rGal27A and rMan113A were galactose and mannose. The yields of mannose, mannobiose, and mannotriose dramatically increased compared with the hydrolysis in the presence of rManA or rMan113A alone.