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As the most abundant renewable carbon source, lignocellulose holds potential as a raw material for biofuels and biochar. The components required for biofuel production differ from those for biochar, so combining processes can reduce costs. Biofuel preparation necessitates cellulase treatment of lignocellulose. This study examines the effects of various enzyme treatment conditions (dosage, time, temperature) on lignocellulose, focusing on the properties of biochar derived from it (BC-SR). A mathematical model was constructed to study the relationship between enzyme treatment conditions and BC-SR properties. BC-SR exhibited high adsorption selectivity for bisphenol A and outperformed untreated biochar in fixed-bed column experiments, demonstrating greater removal efficiency and structural integrity. This study provides insights into the impact of enzymatic treatment on biochar and offers a cost-effective method for producing stable, efficient biochar. Additionally, a highly persistent biochar can enter the carbon trading market as a carbon-neutral technology, further realizing economic and environmental benefits., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Mycelial pellets formed by Penicillium thomii ZJJ were applied as efficient biosorbents for the removal of polycyclic aromatic hydrocarbons (PAHs), which are a type of ubiquitous harmful hydrophobic pollutants. The live mycelial pellets were able to remove 93.48 % of pyrene at a concentration of 100 mg/L within 48 h, demonstrating a maximum adsorption capacity of 285.63 mg/g. Meanwhile, the heat-killed one also achieved a removal rate of 65.01 %. Among the six typical PAHs (pyrene, phenanthrene, fluorene, anthracene, fluoranthene, benzo[a]pyrene), the mycelial pellets preferentially adsorbed the high molecular weight PAHs, which also have higher toxicity, resulting in higher removal efficiency. The experimental results showed that the biosorption of mycelial pellets was mainly a spontaneous physical adsorption process that occurred as a monolayer on a homogeneous surface, with mass transfer being the key rate-limiting step. The main adsorption sites on the surface of mycelia were carboxyl and N-containing groups. Extracellular polymeric substances (EPS) produced by mycelial pellets could enhance adsorption, and its coupling with dead mycelia could achieve basically the same removal effect to that of living one. It can be concluded that biosorption by mycelial pellets occurred due to the influence of electrostatic and hydrophobic interactions, consisting of five steps. Furthermore, the potential applicability of mycelial pellets has been investigated considering diverse factors. The mycelia showed high environmental tolerance, which could effectively remove pyrene across a wide range of pH and salt concentration. And pellets diameters and humic acid concentration had a significant effect on microbial adsorption effect. Based on a cost-effectiveness analysis, mycelium pellets were found to be a low-cost adsorbent. The research outcomes facilitate a thorough comprehension of the adsorption process of pyrene by mycelial pellets and their relevant applications, proposing a cost-effective method without potential environmental issues (heat-killed mycelial pellets plus EPS) to removal PAHs., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Nanobubbles (NBs), given their unique properties, could theoretically be paired with rhamnolipids (RL) to tackle polycyclic aromatic hydrocarbon contamination in groundwater. This approach may overcome the limitations of traditional surfactants, such as high toxicity and low efficiency. In this study, the remediation efficiency of RL, with or without NBs, was assessed through soil column experiments (soil contaminated with phenanthrene). Through the analysis of the two-site non-equilibrium diffusion model, there was a synergistic effect between NBs and RL. The introduction of NBs led to a reduction of up to 24.3 % in the total removal time of phenanthrene. The direct reason for this was that with NBs, the retardation factor of RL was reduced by 1.9 % to 15.4 %, which accelerated the solute replacement of RL. The reasons for this synergy were multifaceted. Detailed analysis reveals that NBs improve RL's colloidal stability, increase its absolute zeta potential, and reduce its soil adsorption capacity by 13.3 %-19.9 %. Furthermore, NBs and their interaction with RL substantially diminish the surface tension, contact angle, and dynamic viscosity of the leaching solution. These changes in surface thermodynamic and rheological properties significantly enhance the migration efficiency of the eluent. The research outcomes facilitate a thorough comprehension of NBs' attributes and their relevant applications, and propose an eco-friendly method to improve the efficiency of surfactant remediation., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier B.V.)

Hepatocellular carcinoma (HCC) is the most prevalent form of primary liver cancer, accounting for 75%-85% of cases. Although treatments are given to cure early-stage HCC, up to 50%-70% of individuals may experience a relapse of the illness in the liver after 5 years. Research on the fundamental treatment modalities for recurrent HCC is moving significantly further. The precise selection of individuals for therapy strategies with established survival advantages is crucial to ensuring better outcomes. These strategies aim to minimize substantial morbidity, support good life quality, and enhance survival for patients with recurrent HCC. For individuals with recurring HCC after curative treatment, no approved therapeutic regimen is currently available. A recent study presented novel approaches, like immunotherapy and antiviral medication, to improve the prognosis of patients with recurring HCC with the apparent lack of data to guide the clinical treatment. The data supporting several neoadjuvant and adjuvant therapies for patients with recurring HCC are outlined in this review. We also discuss the potential for future clinical and translational investigations., Competing Interests: Conflict-of-interest statement: All the authors report no relevant conflicts of interest for this article., (©The Author(s) 2023. Published by Baishideng Publishing Group Inc. All rights reserved.)

Objective: To create the hierarchical model for the comparison of efficacy of different ATs for schizophrenia., Methods: PubMed, Web of Science, Embase, The Cochrane Library, ClinicalTrials, China National Knowledge Infrastructure Database, China Science and Technology Journal Database, Wanfang Database, and SinoMed were searched using a specified search strategy to identify relevant studies up to December 2021. The data were extracted independently by two reviewers. The quality of included trials was evaluated based on the guidelines of "Cochrane Handbook for Systematic Reviews of Interventions". Bayesian network meta-analysis was conducted by statistical analysis software Addis 1.16.6 and Stata 15.1., Results: In total, 60 randomized controlled trials covering 4810 patients were enrolled. The network meta-analysis result showed that Body Acupuncture (BA), BA + Electro-acupuncture (EA), Scalp Acupuncture (SA) + EA, Auricular Acupuncture (AA), Low-dose medication and Acupuncture (LA), Acupoint Injection (AI), and Acupoint Catgut Embedding (ACE), when combined with Western Medications (WM), demonstrated a better clinical effect at improving the symptoms of schizophrenia, compared to WM alone. Results of rank probability showed that BA, when combined with WM, was the most optimal AT for schizophrenia at decreasing three aspects of PANSS scale score., Conclusions: Acupuncture-related therapies help improve the symptoms of schizophrenia, and BA combined with WM may be a better therapy for schizophrenia. This study has been registered on the "PROSPERO" website, and the registration number is CRD42021227403.

Emodin is a natural anthraquinone compound, which is the main component found in the traditional Chinese herb Polygonum cuspidatum. The anti-fibrosis effects of Emodin have been reported. This study aimed to explore the specific mechanism of Emodin in the epithelial-mesenchymal transition (EMT) of pulmonary fibrosis. The pulmonary fibrosis mice models were constructed with bleomycin, the EMT models of alveolar epithelial cells were stimulated by TGF-β1, and Emodin was used for intervention. c-MYC and miR-182-5p were overexpressed or silenced by cell transfection. Our results demonstrated that Emodin attenuated pulmonary fibrosis induced by bleomycin in mice, and inhibited EMT, meanwhile downregulated c-MYC, upregulated miR-182-5p, and downregulated ZEB2 in vitro and vivo. Next, overexpression of c-MYC promoted EMT, while silencing c-MYC and overexpressing miR-182-5p inhibited EMT. Then, c-MYC negatively regulated the expression of miR-182-5p with a direct binding relationship. And miR-182-5p inhibited ZEB2 expression in a targeted manner. Finally, Emodin inhibited EMT that had been promoted by overexpression of c-MYC. In conclusion, Emodin could attenuate pulmonary fibrosis and EMT by regulating the c-MYC/miR-182-5p/ZEB2 axis, which might provide evidence for the application of Emodin in the treatment of pulmonary fibrosis., (© 2022 The Authors. Phytotherapy Research published by John Wiley & Sons Ltd.)

Chronic liver injury eventually progresses to cirrhosis and end-stage liver disease (ESLD), which are the leading causes of death in patients with liver disease worldwide. ESLD has a variety of etiologies and a complex pathogenesis. This study analyzed the characteristics of ESLD by studying the immune microenvironment and inflammatory microenvironment of ESLD caused by 4 noncancer diseases, including HBV-ALF, ALF, AILD, and AH. We collected transcriptome data from noncancer ESLD patients, collected liver tissue samples and blood samples from ESLD liver transplant patients, and analyzed the immune and inflammatory microenvironments in the liver and blood. The results showed that with the exception of HBV-induced ESLD, there were no significant differences in immune microenvironment scores among patients with ESLD caused by other noncancer diseases. Moreover, there were no significant differences in the inflammatory microenvironment in the liver and blood of patients with ESLD caused by the 4 noncancer diseases. Furthermore, we found that the cytokine, IL-15, could predict the prognosis of ESLD patients.

Objective: To investigate the effects of acupotomy on inhibiting abnormal formation of subchondral bone in rabbits with knee osteoarthritis (KOA)., Methods: A total of 24 New Zealand rabbits were randomly divided into four groups of 6 rabbits each [control, model, electroacupuncture (EA) and acupotomy]. Eighteen KOA model rabbits were established using a modified Videman method. Rabbits in EA and acupotomy groups received the intervention for 3 weeks. Then, the cartilage and subchondral bone unit were obtained and the histomorphological changes were recorded. Osteo-protegerin (OPG) and receptor activator of nuclear factor-κB ligand (RANKL) in subchondral bone were evaluated by Western blotting, real-time polymerase chain reaction and immunohistochemistry., Results: Compared with the model group, both the acupotomy and EA groups showed a significant decrease in the Lequesne index (both 0.01) and Mankin score ( 0.01, < 0.05). In addition, both EA and acupotomy groups had a higher expression of total articular cartilage (TAC) ( 0.05, < 0.01) and lower expression of articular calcified cartilage (ACC)/TAC ( 0.05, < 0.05) compared with the model group. The thickness of the subchondral bone plate in EA and acupotomy groups were decreased (both 0.01) compared to the model group. Moreover, trabecular bone volume (BV/TV), protein and relative expression of OPG and the ratio of OPG/RANKL in the subchondral bone of acupotomy group were decreased statistically significant, while these parameters were not significantly changed in the EA group compared with the model group., Conclusions: In the rabbit model of KOA, acupotomy inhibits aberrant formation of subchondral bone by suppressing OPG/RANKL ratio as a potential therapy for KOA.

Background and Objective: Individuals with Alzheimer disease and dementia experience cognitive decline and reduction in physical capabilities. Engaging in cognitive challenges and physical exercises is effective in reducing age-related cognitive and physical decline. It is believed that physical activity in the context of cognitive challenges might enhance the process of neurogenesis in the adult brain, but how effective are such interventions? Is there enough evidence to support that dual-task training is more effective than cognitive or physical training alone? To what extent can such training improve cognitive and physical functions in patients at various stages of cognitive decline?, Methodology: This systematic review with meta-analysis summarizes the emerging evidence of dual-task training for enhancing cognitive and physical functions in older individuals with cognitive impairment, dementia or Alzheimer's disease. A systematic search was carried out in MEDLINE, PubMed, EMBASE, and Cochrane Library with the following search terms: randomized control trials, dual-task training, SCD, MCI, dementia, and Alzheimer's disease., Results: A total of 21 studies with 2,221 participants were identified. The results of dual-task tanning intervention are summarized as change in global cognitive function; SMD = 0.24, (P= 0.002), memory; SMD = 0.28, (P = 0.000), executive function; SMD = 0.35, (P = 0.000), attention; SMD = -0.19, (P = 0.1), gait speed; SMD = 0.26, (P = 0.007), dual-task cost; SMD 0.56, (P = 0.000), and balance; SMD 0.36, (P = 0.004)., Conclusion: Primary analysis showed a small-to-medium positive effect of dual-task training interventions on cognitive functions and medium-to-large positive effect on gait functions and balance., Competing Interests: The authors declare that this research was conducted in the absence of any commercial or financial relationships that could be considered as potential conflict of interest.

The aim of the present study was to investigate the value of the use of fluorine-18 ( 18 F)-fluorodeoxyglucose (FDG) positron emission tomography (PET)/computed tomography (CT) vs. magnetic resonance imaging (MRI) for the diagnosis of and differentiation between benign and malignant lesions in the spinal canal. For this purpose, a retrospective analysis was performed on the use of MRI and 18 F-FDG PET/CT from January, 2017 to December, 2020, and the final diagnosis was obtained by performing a post-operative pathological examination or following a tissue biopsy (gold standard). The sensitivity, specificity, accuracy, positive predictive value and negative predictive value of the two examination techniques were calculated and comparisons between them were made. The PET metabolic parameters, maximum standardized uptake value (SUVmax), peak standardized uptake value (SUVpeak), mean standardized uptake value (SUVmean), metabolic tumor volume (MTV) and total lesion glycolysis (TLG) in the benign and malignant groups were calculated and compared, and the corresponding ROC curves were plotted. A total of 58 patients were enrolled, including 30 patients with malignant and 28 with benign lesions. The specificity of MRI was significantly higher than that of PET/CT (P<0.05). The sensitivity and negative predictive value of PET/CT were higher than those of MRI, although with no significant difference (P>0.05). The mean ± tandard deviation values of the PET metabolic parameters, SUVmax, SUVpeak, SUVmean, MTV and TLG, were 4.27±1.25, 3.49±1.07, 2.49±0.84, 6.58±5.36 and 17.12±15.50 in the benign, and 8.99±3.75, 7.35±3.26, 5.43±2.40, 12.25±12.18 and 112.41±85.98 in the malignant groups, respectively. The SUVmax, SUVpeak, SUVmean and TLG in the malignant group were higher than those in the benign group. The differences were all statistically significant (all P<0.0001). In distinguishing benign from malignant lesions, the area under the ROC curve (AUC) for SUVmax was 0.919, which was the largest, and the Youden index was 0.762, indicating 83.3% sensitivity and 92.9% specificity. The AUC for SUVpeak was 0.905 and that for SUVmean was 0.899. The aforementioned AUCs were significantly higher than those for MTV and TLG (0.609 and 0.786, respectively) (P<0.001). On the whole, the present study demonstrates that MRI is a reliable imaging technique for the diagnosis of intravertebral lesions. 18 F-FDG PET/CT, as a noteworthy supplement to MRI, has a high sensitivity and accuracy for the qualitative diagnosis and identification of lesions. The synergistic effect of the two examination techniques may be helpful for a more accurate diagnosis., Competing Interests: The authors declare that they have no competing interests., (Copyright: © Zheng et al.)

Objective: To explore the interventional therapy and clinical efficacy of extracranial ICA aneurysm., Methods: The clinical data of eight patients with extracranial ICA aneurysm treated by interventional stent implantation from December 2014 to February 2018 in the Neurosurgery Department of the Third Hospital of Mianyang were analyzed. And this research was a retrospective analysis. All patients underwent digital subtraction angiography (DSA) and were diagnosed with extracranial carotid artery aneurysm. These patients, therefore, were treated with interventional stent implantation., Results: Interventional treatment was successfully conducted on all eight patients. In eight patients, the aneurysm cavity was not developed immediately after angiography, and in one case, the aneurysm cavity was developed with coil-assisted embolization. All the internal carotid arteries were well developed, with no complications such as intraoperative rupture, bleeding and thrombosis occur. Follow-up for three months to two years showed that the patients recovered well, the GOS score was 4 points for patients with cerebral infarction, and the rest reached five points. Follow-up CTA showed no signs of aneurysm recurrence or ICA restenosis., Conclusion: Interventional stent placement is a preferable and relatively safe method for the treatment of extracranial carotid artery aneurysm with less trauma and short operation time., Competing Interests: Conflicts of interest: None, (Copyright: © Pakistan Journal of Medical Sciences.)

Bisphenol A (BPA) is a widely used raw material that can be detected both in the environment and in the human body. Due to its estrogen-like effects, wide concerns have been raised about the potential role of BPA in the initiation and development of hormone-dependent cancers. Ovarian cancer is the most common reproductive system cancer and has a high mortality rate in women. Despite recent investigations into BPA's carcinogenic effects, studies on its role in ovarian cancer development remain limited. In this study, we aimed to assess the effect of BPA at various environmentally relevant concentrations on proliferation and metastasis of ovarian cancer cells. We discovered that BPA can stimulate proliferation of OVCAR-3 ovarian cancer cells after exposure for up to 5 days. Strikingly, BPA enhanced ovarian cancer cell migration, invasion, and adhesion (to vascular endothelial cells) through upregulation of matrix metalloproteinase-2 (MMP-2), MMP-9, and intercellular cell adhesion molecule-1 (IMAC-1). The stimulatory effects of BPA on cancer cell proliferation and metastasis were reversed by treatment with an ERα inhibitor, but not by treatment with an ERβ inhibitor. Together, these results suggest that BPA induces proliferation and metastasis of ovarian cancer cells through ERα signaling pathways. This study provides new insights into the carcinogenic effects of BPA with regard to ovarian cancer., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Objective: To investigate the correlation between monocyte to high-density lipoprotein ratio (MHR) and major adverse cardiovascular events (MACE) in patients with acute coronary syndrome (ACS) after percutaneous coronary intervention (PCI)., Methods: In this retrospective study, 120 ACS patients who received PCI in our hospital from September 2014 to August 2019 were selected and divided into MACE group and normal discharge (ND) group. Their clinical data were collected, and MHR values were compared. Logistic regression analysis was conducted to analyze the correlations between various factors and ACS. The correlation between MHR and Gensini score was subjected to Pearson's analysis. Receiver operating characteristic (ROC) curve was plotted to analyze the diagnostic value of MHR for MACE., Results: Hypertension degree, white cell count, Gensini score, MHR and the levels of total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDLC), high-density lipoprotein cholesterol (HDLC), apolipoprotein A1 (ApoA1), ApoB, lipoprotein (a) [LP(a)] and uric acid (UA) in MACE group were significantly higher than those in ND group (P<0.05). HDLC, ApoA1, TC, MHR, LDLC and ApoB were independent risk factors for MACE of ACS patients after PCI (P<0.05). There was a positive correlation between MHR and Gensini score (r=0.832, P<0.05), and the optimal cutoff value of MHR for diagnosing MACE was 9.45., Conclusion: Serum MHR is positively correlated with Gensini score in ACS patients after PCI, which can be used as an independent predictor for MACE in hospital., Competing Interests: Conflict of interest: None., (Copyright: © Pakistan Journal of Medical Sciences.)

Tumor biomarkers are important in the early screening, diagnosis, therapeutic evaluation, recurrence and prognosis prediction of tumors. Primary liver cancer is one of the most common malignant tumors; it has high incidence and mortality rates and seriously endangers human health. The main pathological types of primary liver cancer include hepatocellular carcinoma (HCC), intrahepatic cholangiocarcinoma (ICC) and combined HCC‑cholangiocarcinoma (cHCC‑CC). In the present review, a systematic outline of the current biomarkers of primary liver cancer is presented, from conventional blood biomarkers, histochemical biomarkers and potential biomarkers to resistance‑associated biomarkers. The important relationships are deeply elucidated between biomarkers and diagnosis, prognosis, clinicopathological features and resistance, as well as their clinical significance, in patients with the three main types of primary liver cancer. Moreover, a summary of several important biomarker signaling pathways is provided, which is helpful for studying the biological mechanism of liver cancer. The purpose of this review is to provide help for clinical or medical researchers in the early diagnosis, differential diagnosis, prognosis and treatment of HCC.

Background: Acute cerebral infarction (ACI) is seriously harmful to human health worldwide. However, at present, the risk of disease onset is still not accurately predicted for some people., Methods: Five hundred and nineteen patients with ACI and 300 healthy controls were included in this study. We divided the patients into three groups according to the results of cervical artery contrast-enhanced ultrasound. Ninety-five patients were in the CAS without plaque group, 108 patients were in the stable plaque group, and 316 patients were in the unstable plaque group. TC, TG, HDL-C, LDL-C, and sdLDL-C were measured in all subjects., Results: The level of small dense low-density lipoprotein cholesterol (sdLDL-C) in the ACI group was significantly higher than that in the control group (P < 0.001). Logistic regression analysis showed that sdLDL-C was an independent risk factor for ACI (OR = 1.067, 95% CI: 1.041-1.093, P < 0.001); serum sdLDL-C was significantly higher in the unstable plaque group than in the stable plaque group and plaque-free group (P < 0.05, P < 0.001); serum sdLDL-C was also higher in the stable plaque group than the plaque-free group (P < 0.001). Logistic regression analysis showed that sdLDL-C was an independent risk factor for unstable carotid plaques (OR = 1.053, 95% CI: 1.038-1.068, P < 0.001); Spearman correlation analysis showed that sdLDL-C test results were positively correlated with carotid plaque stability (r = 0.363, P < 0.001)., Conclusion: Small dense low-density lipoprotein cholesterol is an independent risk factor for the onset of ACI and may be an early serum marker for this disease., (© 2019 The Authors. Journal of Clinical Laboratory Analysis Published by Wiley Periodicals, Inc.)

Steroids, including testosterone, estrone, 17β-estradiol, estriol and 17β-ethinyl estradiol, are harmful not only to the population dynamics of aquatic life forms but also to public health. In this study, a marine testosterone-degrading bacterium (strain N3) was isolated from Nanao Island in the South China Sea. In addition, the strain could also use 17β-estradiol (E2), 17β-ethinyl estradiol (EE2), estriol (E3) or cholesterol as a sole carbon source. According to the 16S rRNA gene sequence analysis, strain N3 was identified as Vibrio sp. Further characterization showed that the strain is aerobic, gram-negative, and mobile and exhibits resistance to ampicillin, carbenicillin, penicillin and spectinomycin. For enhancing its capacity of testosterone degradation, the Plackett-Burman factorial design and the central composite design were used to optimize the culture condition. Under optimal conditions, 92% of testosterone was degraded by Vibrio sp. N3 in 48h., (Copyright © 2018. Publicado por Elsevier España, S.L.U.)

Continuous blood pressure (BP) monitoring has a significant meaning to the prevention and early diagnosis of cardiovascular disease. However, existing continuous BP monitoring approaches, especially cuff-less BP monitoring approaches, are all contraptions which complex and huge computation required. For example, for the most sophisticated cuff-less BP monitoring method using pulse transit time (PTT), the simultaneous record of photoplethysmography (PPG) signal and electrocardiography (ECG) are required, and various measurement of characteristic points are needed. These issues hindered widely application of cuff less BP measurement in the wearable devices. In this study, a novel BP estimation method using single PPG signal feature was proposed and its performance in BP estimation was also tested. The results showed that the new approach proposed in this study has a mean error -0.91 ± 3.84 mmHg for SBP estimation and -0.36 ± 3.36 mmHg for DBP estimation respectively. This approach performed better than the traditional PTT based BP estimation, which mean error for SBP estimation was -0.31 ± 4.78 mmHg, and for DBP estimation was -0.18 ± 4.32 mmHg. Further investigation revealed that this new BP estimation approach only required measurement of one characteristic point, reducing much computation when implementing. These results demonstrated that this new approach might be more suitable implemented in the wearable BP monitoring devices.

In this study, the cells-free and cells-loaded chitosan hydrochloride-alginate (CHC-Alg) microcapsules were firstly fabricated with polyelectrolyte complexes via an orifice-polymerization method. Scanning electron microscope images showed that the CHC-Alg microcapsules had a typical shell-core structure and the model probiotic cells (Bacillus licheniformis) were embedded in the core in cells-loaded microcapsules. The microcapsules prepared had good thermal stability and moisture property (3.89%). Cells survival and release studies showed that the number of probiotic cells released from the cells-loaded microcapsules (approx. 6.36logCFUml -1 ) was 6.19logCFUml -1 when they were performed in the simulated gastric fluid (SGF, pH 2.0) for 1h and subsequently in the simulated intestinal fluid (SIF, 0.3%) for 4h. The CHC-Alg microcapsules with favorable swelling performances were helpful to permeate the harsh acid to protect the cells in the SGF (pH 2.0). The CHC-Alg microcapsules effectively protected the model probiotic cells, which was attributed to the "dual protective barriers" of the shell-core structure, that is, the primary barrier of the Alg hydrogel layer formed with a compact polymer matrix and the secondary barrier of the PEC film formed on the surface. The microcapsules prepared could be used as an enteric micro-probiotic-carrier for designing potential probiotic delivery systems., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Aim: To identify and assess the research situation of top 100 cited articles in nonalcoholic fatty liver disease (NAFLD)., Methods: The global scientific research articles in the Science Citation Index-Expanded relevant to NAFLD were retrieved and listed according to their citation times from the most to the least. The 100 most frequently cited original articles were selected to systematically evaluate their bibliometric parameters including times cited, publication year, journals, subject categories, and the highly related concepts of NAFLD, which reflected the history and current situation, publication distribution of leading countries and institutes as well as the research hotspots of NAFLD., Results: Top 100 cited articles in NAFLD were published from 1965 to 2015 with a citation ranging of 227 to 2151 times since publication, in which the United States was the most predominant country and Mayo Clin was the most productive institution. The majority of the top 100 cited articles were concentrated in SCI subject category of Gastroenterology and Hepatology. Hepatology and Gastroenterology is the top journal that published over half 100 top-cited articles. The significant peak of top cited articles present in the first half of the 2000s while the highest mean number of citation presents in first half of the 1980s. In addition, concepts related to pathology characteristics, epidemiology and medicalization, metabolic syndrome and its combination of symptoms including insulin resistance, biomarkers of lipid metabolism and obesity are listed as the highly related concepts., Conclusion: The 100 top-cited articles marked with the leading countries, institutions, journals, hotspots and development trend in NAFLD field that could provide the foundation for further investigations., Competing Interests: Conflict-of-interest statement: No conflicts of interest, financial or otherwise, are declared by the authors.

Based on the properties of oppositely charged natural polysaccharides, the polyelectrolyte complexes (PECs) prepared with chitosan-related polycationic polyelectrolytes and cellulose-related polyanionic polyelectrolytes have been widely concerned for their potential applications as micro-drug-carriers for colon. However, the poor mechanical property of the PECs becomes the obstacle encountered in practical applications. This study investigated the effect of the cross-linking agent (sodium polyphosphate, PPS) on the performances of sodium cellulose sulfate -chitosan/water soluble chitosan (NaCS-CS/WSC) microcapsules. The results revealed that PPS could penetrate through the PEC film and form tighter interior structures compared with the microcapsules without the addition of cross-linking agent. The NaCS-CS microcapsules and NaCS-WSC microcapsules with or without PPS had distinct microstructures, which could be ascribed to the different physicochemical properties of CS and WSC. During the formation process, CS can be dissolved in water under acidic conditions, while WSC can be directly dissolved and protonated in acid-free aqueous providing NH3(+) groups quickly, which resulted in the microstructure's difference. Further analysis showed the NaCS-CS-PPS microcapsules and NaCS-WSC-PPS microcapsules had lower swelling ratios due to their tighter interior microstructures that formed. The cross-linking agent had important effect on the total mass of PECs that produced; moreover, the decline of zeta potential of NaCS-CS-PPS microcapsules was lower than that of NaCS-CS microcapsules, similar trend was found in the NaCS-WSC-PPS microcapsules compared with NaCS-WSC microcapsules, indicating the PPS participated in the interactions and played a role in the microcapsules' formation process., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Objective: This study investigated the influence of Hawley retainer and vacuum-formed retainer on the articu-lation of Chinese speech of adult orthodontic patients., Methods: A total of 20 volunteers were recruited in this randomized controlled trial in accordance with the inclusion criteria. Participants were randomly distributed into two groups, namely, the Hawley retainer group and the vacuum-formed retainer group. Speech performance was evaluated at the following time inter-vals: before wearing retainers (T0), immediately after wearing both upper and lower retainers (T1), 24 h later (T2), 1 week later (T3), and 1 month later (T4). The phonetic parameters were acquired through the use of computer-based voice analysis software Praat version 5.4.21, and the final data were analyzed using SPSS 19.0., Results: Both retainers can influence the phonetic parameters of vowels /e/, /i/, and /ü/ and consonants /s/, /f/, /x/, /t/, /j/, and /q/. However, the Hawley retainer showed a more remarkable effect on the articulation of Chinese speech, and the distortion of /i/, /ü/, and /s/ sounds can last for more than 1 month. Nevertheless, the recovery period of the patients' phonetic function in the vacuum-formed retainer group was within 1 month., Conclusions: Both retainers can induce distortion of some Chinese vowels and consonants in the speech of adult orthodontic patients, but the Hawley retainer can more significantly influence the articulation of Chinese speech than vacuum-formed retainer.
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To identify the fuctions of the nine putative O-methyltransferase genes in Streptomyces virginiae IBL14, the evolutionary and functional relationship of these genes in its 8.0 Mb linear chromosome was set up via sequence comparison with those of other Streptomyces species. Further, the functions and catalytic mechanism of the three genes sviOMT02, sviOMT03 and sviOMT06 from this strain were studied through experimental and computational approaches. As a result, the nine putative O-methyltransferases belong to methyltransf&#95;2 superfamily, amdomet-MTases superfamily, and leucine carboxyl methyltransferase superfamily, and are phylogenetically close to those of Streptomyces sp. C. The products of genes sviOMT03 and sviOMT06 could catalyze O-methylation of caffeic acid to form ferulic acid. Computational analysis indicated that the O-methylation mechanism of SviOMT03 and SviOMT06 proceeds from a direct transfer of the SAM-methyl group to caffeic acid with inversion of symmetry aided by a divalent metal ion in a SN2-like mechanism. Particularly, the conservative polar amino acid residues in SviOMT03 and SviOMT06, including Lys143 that reacts with caffeic acid, Ser74, Asp140 and Tyr149 that react with S-adenosyl methionine, and His142 (SviOMT03) or His171 (SviOMT06) that transfers the 3-hydroxyl proton of substrate caffeic acid, probably be essential in their O-methylation., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Chronic rhinosinusitis (CRS) is a common disease worldwide, with a prevalence rate of 5%-15% in the general population. CRS is currently classified into two types: CRS with and without nasal polyps. CRS may also be divided into eosinophilic CRS (ECRS) and non-ECRS subtypes based on the presence of tissue eosinophilic infiltration or not. There are significant geographic and ethnic differences in the tissue eosinophilic infiltration, which is predominant in Western white patients and less common in East Asians, despite an increasing tendency for its prevalence in East Asia countries. ECRS differs significantly from non-ECRS in clinical characteristics, treatment outcomes and strategies, and underlying pathogenic mechanisms. ECRS commonly demonstrates more severe symptoms, polyp diseases with a higher incidence of bilateral polyps and sinonasal diseases on computed tomography, and the increase in blood eosinophils. ECRS is considered a special and recalcitrant subtype of CRS, commonly with poor treatment outcomes compared to non-ECRS. The differentiation of specific subtypes and clinical features of CRS will be important for developing novel treatment strategies and improving treatment outcomes for individual phenotypes of CRS. This review discusses clinical features, diagnosis, treatment and prognosis of ECRS in East Asians.

In this study, a strategy of the construction of leaky strains for the extracellular production of target proteins was exploited, in which the genes mrcA, mrcB, pal and lpp (as a control) from Escherichia coli were knocked out by using single- and/or double-gene deletion methods. Then the recombinant strains for the expression of exogenous target proteins including Trx-hPTH (human parathyroid hormone 1-84 coupled with thioredoxin as a fusion partner) and reteplase were reconstructed to test the secretory efficiency of the leaky strains. Finally, the fermentation experiments of the target proteins from these recombinant leaky strains were carried out in basic media (Modified R media) and complex media (Terrific Broth media) in flasks or fermenters. The results demonstrated that the resultant leaky strains were genetically stable and had a similar growth profile in the complex media as compared with the original strain, and the secretory levels of target proteins into Modified R media from the strains with double-gene deletion (up to 88.9%/mrcA lpp-pth) are higher than the excretory levels from the strains with single-gene deletion (up to 71.1%/lpp-pth) and the host E. coli JM109 (DE3) (near zero). The highest level of extracellular production of Trx-hPTH in fermenters is up to 680 mg l(-1)., (© 2014 The Authors. Microbial Biotechnology published by John Wiley & Sons Ltd and Society for Applied Microbiology.)

Background: For a therapeutic intervention after spinal cord injury (SCI), it is important to take accurate and objective assessment tools., Objective: To explore the practicability of somatosensory evoked potentials (SEPs) and Modified Barthel Index (MBI) scale and describe the rehabilitation value of SEPs in different degrees of SCI., Methods: Thirty-six SCI patients were enrolled in this study. All the patients received comprehensive rehabilitation treatment, such as physical therapy, occupational therapy, functional electrical stimulation, and psychotherapy. The nerve function of the spinal cord was assessed by SEPs, the activities of daily living (ADL) was evaluated by MBI scale, and SEP recordings and MBI scores were obtained before and after treatment., Results: There were statistically significant differences in SEPs latency among different grades of SCI before treatment. The SEPs latency after treatment was better than that before treatment in every grade (p < 0.05). Comparable differences among different grades were also detected by MBI scores before treatment (p < 0.05), and the MBI scores increased significantly after treatment (p < 0.05), higher in each group than another from grade A to B, C, and D. There was a linear correlation between SEPs latency and MBI scores before and after treatment., Conclusion: SEPs combined with MBI scale could objectively reflect the SCI degree and accurately monitor therapeutic intervention in SCI. SEPs have a greater value in monitoring SCI than MBI and their rehabilitation value varies in different grades of SCI.

Background: As well known, both natural and synthetic steroidal compounds are powerful endocrine disrupting compounds (EDCs) which can cause reproductive toxicity and affect cellular development in mammals and thus are generally regarded as serious contributors to water pollution. Streptomyces virginiae IBL14 is an effective degradative strain for many steroidal compounds and can also catalyze the C25 hydroxylation of diosgenin, the first-ever biotransformation found on the F-ring of diosgenin., Results: To completely elucidate the hydroxylation function of cytochrome P450 genes (CYPs) found during biotransformation of steroids by S. virginiae IBL14, the whole genome sequencing of this strain was carried out via 454 Sequencing Systems. The analytical results of BLASTP showed that the strain IBL14 contains 33 CYPs, 7 ferredoxins and 3 ferredoxin reductases in its 8.0 Mb linear chromosome. CYPs from S. virginiae IBL14 are phylogenetically closed to those of Streptomyces sp. Mg1 and Streptomyces sp. C. One new subfamily was found as per the fact that the CYP Svu001 in S. virginiae IBL14 shares 66% identity only to that (ZP&#95;05001937, protein identifer) from Streptomyces sp. Mg1. Further analysis showed that among all of the 33 CYPs in S. virginiae IBL14, three CYPs are clustered with ferredoxins, one with ferredoxin and ferredoxin reductase and three CYPs with ATP/GTP binding proteins, four CYPs arranged with transcriptional regulatory genes and one CYP located on the upstream of an ATP-binding protein and transcriptional regulators as well as four CYPs associated with other functional genes involved in secondary metabolism and degradation., Conclusions: These characteristics found in CYPs from S. virginiae IBL14 show that the EXXR motif in the K-helix is not absolutely conserved in CYP157 family and I-helix not absolutely essential for the CYP structure, too. Experimental results showed that both CYP Svh01 and CYP Svu022 are two hydroxylases, capable of bioconverting diosgenone into isonuatigenone and β-estradiol into estriol, respectively.

Background: To evaluate the efficacy of combined treatment with bone marrow mesenchymal stem cell (BMSC) transplantation and methylprednisolone (MP) to treat paraquat (PQ)-induced acute lung injury., Materials and Methods: A total of 102 female rats were randomly divided into five groups: PQ, BMSC, MP, BMSC + MP and normal control. After 14 days of PQ poisoning, the survival of rats, wet/dry weight ratio of lung tissue, serum levels of tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-6, IL-10, malondialdehyde (MDA) and superoxidase dismutase (SOD), and the expression of nuclear factor (NF)-кB p65 in lung tissue were determined., Results: Rats in BMSC and BMSC + MP groups survived. BMSC transplantation significantly decreased the wet/dry weight ratio of lung tissue, down-regulated NF-кB p65 expression in lung tissue, lowered serum levels of TNF-α, IL-1β, IL-6 and MDA, and increased serum levels of IL-10 and SOD. These changes were particularly significant on days 7-14 after PQ poisoning. The above changes were more significant in the MP group on days 1-3 after PQ poisoning, compared with those of the BMSC group. However, the BMSC + MP group showed more significant changes on days 1-14 after PQ poisoning than those of both BMSC and MP groups., Conclusions: MP inhibits the inflammatory response, reduces the products of lipid peroxidation and promotes survival of transplanted BMSC, thus improving the intermediate and longer term efficacy of BMSC transplantation for treatment of PQ-induced lung injury.

Background: Sclerosing hemangioma (SH) of the lung is a kind of rare pulmonary tumor. Preoperative diagnosis of this tumor is difficult and it is now generally accepted that SH of the lung is benign lesions and surgical excision alone is curative. Herein, we present our experiences of treating 28 patients with SH., Methods: The medical records of 28 patients with SH from 1994 to 2010 at the Department of Thoracic Surgery in Beijing Chest Hospital were retrospectively reviewed., Results: There were 3 male and 25 female patients with sclerosing hemangioma and 50% of the patients were asymptomatic. Preoperatively, all the patients had undergone CT of chest and 5 patients had undergone PET scan but 4 patients were misdiagnosed as malignancy. There was no operative mortality or tumor recurrence despite that three different operative methods were undertaken., Conclusions: SH has a high incidence in middle-aged women. Most of SH is asymptomatic and the symptoms of SH are not related to the tumor size and distribution. The features of chest CT and PET are not specific. Bilateral or multiple lesions should not exclude the possibility of SH. Complete excision of lesion is a curable treatment method and there is no evidence to verify the need of adjuvant therapy.

Acute lung injury (ALI) and acute respiratory distress syndrome (ARDS) are associated with high rates of morbidity and mortality. Currently, several surfactant or anti-inflammatory drugs are under test as treatments for ALI. Sodium aescinate (SA) has been shown to exert anti-inflammatory and antiedematous effects. In the present work, the authors explored the effects of SA and the possible mechanisms of SA action in rats with ALI induced by oleic acid (OA) administration. Eight groups of rats received infusions of normal saline (NS) or OA. Rats exposed to OA were pretreated with 1 mg/kg of SA, or posttreated with SA at low (1 mg/kg), medium (2 mg/kg), or high (6 mg/kg) dose; a positive-control group received methylprednisolone. The pressure of oxygen in arterial blood (P(O(2))) levels, the pulmonary wet/dry weight (W/D) ratios, and indices of quantitative assessment (IQA) of histological lung injury were obtained 2 or 6 hours after OA injection (0.1 mL/kg, intravenously). The levels of superoxide dismutase (SOD), malondialdehyde (MDA), matrix metalloproteinase gelatinase B (MMP-9), and tissue inhibitor of metalloproteinase (TIMP-1) in both plasma and lung tissue were also determined. Both pre- and posttreatment with SA improved OA-induced pulmonary injury, increased P(O(2)) and SOD values, lowered IQA scores, and decreased the lung W/D ratio and MDA and MMP-9 levels in plasma and lung tissue. SA appeared to abrogate OA-induced ALI by modulating the levels of SOD, MDA, and MMP-9 in plasma and lung tissue.

Objective: To comprehensively summarize the associations between retinol, vitamins A, C, and E and breast cancer, and quantitatively estimate their dose-response relationships., Methods: We searched PubMed, Embase, and Cochrane databases (from January 1982 to 15 March 2011) and the references of the relevant articles in English with sufficient information to estimate relative risk or odds ratio and the 95% confidence intervals, and comparable categories of vitamins. Two reviewers independently extracted data using a standardized form, with any discrepancy adjudicated by the third reviewer., Results: Overall, 51 studies met the inclusion criteria. Comparing the highest with the lowest intake, total vitamin A intake reduced the breast cancer risk by 17% (pooled OR = 0.83, 95% CI: 0.78-0.88). Further subgroup analysis based on study design did not change the significant reduction. Although the dietary vitamin A, dietary vitamin E, and total vitamin E intake all reduced breast cancer risk significantly when data from all studies were pooled, the results became nonsignificant when data from cohort studies were pooled. The significant association between total retinol intake and breast cancer in all studies became nonsignificant in case-control studies but remain significant in cohort studies. No significant dose-response relationship was observed in the higher intake of these vitamins with reduced breast cancer risk., Conclusions: Our results indicate that both the total intake of vitamin A and retinol could reduce breast cancer risk. However, associations between other vitamins and breast cancer seem to be limited.

Objective: How to organise an appropriate team to provide quality dental care during the Olympic Games has become an important consideration for each successive host country. The aims of this study were to document dental services provided at the Olympic Games and to provide data for planning future events. SET-UP OF THE DENTAL CARE DEPARTMENT: There were six dental chairs in six independent treatment rooms, one technical laboratory, a sterilising room and an x-ray room equipped with one digital panoramic screening machine and one intraoral x-ray machine in the polyclinic in the Olympic Village in Beijing. Shifts comprised 80 dentists and 28 nurses who were organised into three shifts working from 08:00 until 23:00., Results: In the 2008 Olympic Games, there were 1607 cases involving 1126 patients requiring dental care: 795 cases from 516 athletes; 483 cases from 370 coaches and other staff; and 99 cases from volunteers. Endodontic treatments, permanent fillings, oral hygiene, mouthguards and treatment of pericoronitis were the most frequent procedures in dental care. The Mouthguard Service was extremely popular and well utilised. 122 athletes received new custom-made mouthguards in Beijing., Conclusions: As the utilisation of the dental service grows, and the burden of providing care for such a large cohort increases, a well-organised dental team becomes increasingly important. More general-practice dentists were needed in the team. Different kinds of specialist were suggested for working in the team-for example, oral and maxillofacial surgeons, sport dentists for mouthguards and endodontists.

OBJECTIVE. To examine the protective effect of bone marrow mesenchymal stem cell (BMSC) transplantation against lung injury induced by paraquat (PQ) poisoning. MATERIALS AND METHODS. Fifty-four female SD rats were randomly divided into four groups: a PQ group, a BMSC treatment group, a BMSC control group, and a normal control group. BMSC, isolated and cultured in vitro, were injected into the tail veins of the rats. The mortality rate, the ratio of lung wet/dry weight, plasma levels of IL-1β, tumor necrosis factor-α (TNF-α), malonaldehyde (MDA), superoxide dismutase (SOD), glutathione peroxidase (GSH-P(X)), and NF-κBp65 expression in the lung tissues were examined during 14 days after injection. Results. Survival in the PQ group ranged from 8 to 12 days, with an average of 9.6 days. All the rats in the BMSC treatment group survived for more than 14 days. In the BMSC treatment group, the ratio of lung wet /dry weight (P  =  0.02  <  0.05), plasma IL-1β and TNF-α levels (P  =  0.00  <  0.01), MDA levels (P  = 0.03  <  0.05), and expression of NF-κBp65 in the lung tissues (P  =  0.00  <  0.01) were significantly lower than those in the PQ group by day 7 post-injection. However, the SOD and GSH-P(X) levels began to increase (P  =  0.00  < 0.01) compared with the PQ group. There was no significant difference between the BMSC control group and the normal control group (P  >  0.01). DISCUSSION AND CONCLUSION. BMSC transplantation may have the potential to protect against PQ-induced lung injury, by reducing lung edema and lipid peroxidation, inhibiting the release of inflammatory mediators. However, the protective effects require time to develop, and early BMSC transplantation may be more evident during the middle and late stages of therapy.

Aim: To assess neovascularization within human carotid atherosclerotic soft plaques in patients with ischemic stroke., Methods: Eighty-one patients with ischemic stroke and 95 patients without stroke who had soft atherosclerotic plaques in the internal carotid artery were studied. The thickest soft plaque in each patient was examined using contrast-enhanced ultrasound. Time-intensity curves were collected from 5 s to 3 min after contrast injection. The neovascularization within the plaques in the internal carotid artery was evaluated using the ACQ software built into the scanner by 2 of the experienced investigators who were blinded to the clinical history of the patients., Results: Ischemic stroke was present in 7 of 33 patients (21%) with grade I plaque, in 14 of 51 patients (28%) with grade II plaque, in 26 of 43 patients (61%) with grade III plaque, and in 34 of 49 patients (69%) with grade IV plaque (P < 0.001 comparing grade IV plaque with grade I plaque and with grade II plaque and P = 0.001 comparing grade III plaque with grade I plaque and with grade II plaque). Analysis of the time intensity curves revealed that patients with ischemic stroke had a significantly higher intensity of enhancement (IE) than those without ischemic stroke (P < 0.01). The wash-in time (WT) of plaque was significantly shorter in stroke patients (P < 0.05). The sensitivity and specificity for IE in the plaque were 82% and 80%, respectively, and for WT were 68% and 74%, respectively. There was no significant difference in the peak intensity or time to peak between the 2 groups., Conclusion: This study shows that the higher the grade of plaque enhancement, the higher the risk of ischemic stroke. The data suggest that the presence of neovascularization is a marker for unstable plaque.

A bacterium Streptomyces virginiae IBL-14 capable of effective degradation of diosgenin was isolated from activated sludge for treatment of waste from a steroidal drug factory. From the culture broth of diosgenin degradation, 11 compounds were purified and then identified, eight of which were previously unidentified compounds including 1-dehydroisonuatigenone [VI], nuatigenone [VIII], 1-dehydronuatigenone [X], 26-acetyl-nuatigenone [XII], 6-methoxy-6-dehydrodiosgenone [XIII], 6-methoxy-6-dehydroisonuatigenone [XIV], 6-methoxy-6-dehydronuatigenone [XV], and 6-dimethoxy-7alpha-hydroxyldiosgenone [XVI]. Additionally, two important microbial transformations of diosgenin (6-methoxylation and C25-tertiary carbon hydroxylation) were found. Two valuable chemical reactions of the steroids (structural rearrangement and esterification)were also confirmed. As a result, a new metabolic pathway of diosgenin metabolism was postulated.

About 300 of steroid drugs have been identified since the research and development to steroid drugs was triggered in 1950's. Although much progress in microbial biotransformation on steroid drugs is well-achieved, many efforts are ongoing in order to improve the efficiency of steroidal drug production as well as to degrade their derivatives. In this work an overview of recent and important findings related to patents and various microbial biotransformations of steroidal compounds including dehydrogenation/reduction, hydroxylation, esterification, methylation, halogenation and methoxylation is presented. Both metabolic pathway and genetic manipulation and screening for new strains capable of metabolizing/transforming steroid drugs, as the powerful endocrine disruptors, are introduced since they are serious environmental contaminants.

To improve the plasmid stability during the production of antihuman ovarian carcinoma x antihuman CD3 single-chain bispecific antibody (AhOCxAhCD3), the Escherichia coli BL21(DE3) host cell was optimized serially. Firstly, an isogenic recombination-deficient (recA (-)) derivative of BL21(DE3), namely BLR(DE3), was used as host instead of BL21(DE3). Although the segregational plasmid stability was greatly improved, AhOCxAhCD3 yield was not improved due to the severe growth inhibition of plasmid-bearing BLR(DE3) cells and the competitive plasmid instability after induction. Secondly, a mutant BLR(DE3), namely BLRM(DE3), was screened by using LB agar plates plus ampicillin and isopropyl-beta-D: -thiogalactopyranoside. Using this new host, growth inhibition of recombinant cells after induction was eliminated, and plasmids could be stably maintained even after long-time induction in a nonselective medium. At last, about 1.2 g/l AhOCxAhCD3, which was about thrice as much as those of recombinant BL21(DE3) and BLR(DE3) strains, was yielded.

Formation of inclusion bodies is an important obstacle to the production of active recombinant protein in Escherichia coli. Thus, soluble expression of penicillin G acylase from Kluyvera citrophila was investigated in BL21(DE3). In this study, the yield of active enzyme was significantly enhanced by the composition of the medium and induction opportunity. When 0.5 mmol/L IPTG was added to complex medium at 15 h after incubation, the volumetric and specific activities of penicillin G acylase both achieved the highest values, respectively. However, aggravation of intracellular proteolysis and decline of enzyme expression were also observed if induction occurred too much later. Ca2+ ion was another critical factor in cell growth and protein expression. When 24 mmol/L Ca2+ ion was adding to the medium at the beginning of fermentation, a greater than 2-fold increase in cell density and a 7-fold increase in volumetric activity of penicillin G acylase were reached. Nevertheless, no significant benefit for recombination protein expression was found when excess Ca2+ was added after induction time. This study demonstrates that the induction starting time and Ca2+ ion are two critical factors for the expression of active penicillin G acylase.

To obtain high-quality AhOC x AhCD3 [anti-(human ovarian carcinoma)xanti-(human CD3)] with high biological activity economically and easily, some characteristics and purification of AhOC x AhCD3, a single chain bispecific antibody, were investigated. During the present study, some important properties of AhOC x AhCD3, such as molecular mass, pI, solubility, stability, hydrophobic ability, molecular status and bioactivity were primarily studied. The molecular mass of AhOC x AhCD3 was determined to be approx. 58.0 kDa by SDS/PAGE (theoretical molecular mass: 56972.5 Da, calculated on the basis of the primary structure) and the pI was approx. 7.5+/-0.4 by IEF (isoelectric focusing; theoretical pI: 8.5, predicted by the ProtParam software tool) respectively. Experiments showed that the solubility of AhOC x AhCD3 increased with the pH increase and that the AhOC x AhCD3 was easily degraded into several fragments during the storage time of samples and the mixtures of both fragment B and fragment C retained normal bioactivity. Particularly, the soluble aggregate/polymer status of AhOC x AhCD3 with bioactivity was verified by non-reducing SDS/PAGE. After optimization of purification process, AhOC x AhCD3 with electrophoretic homogeneity was successfully obtained and the yield was approx. 20%. Some important properties of AhOC x AhCD3 were first observed and a procedure for soluble AhOC x AhCD3 purification in scale-up was first established. Some characteristics of AhOC x AhCD3, especially the stability of AhOC x AhCD3, are critical for the development of this protein pharmaceutical and useful for reference to other proteins.

A modified purification method, thermoosmotic shock (osmotic shock coupled with heat-treatment) for heat-stable proteins, was devised in the purification of Trx-hPTH (1-84) (human parathyroid hormone coupled with thioredoxin as a fusion partner) from E. coli. Thermoosmotic shock can integrate the functions of extraction and crude separation of fusion protein Trx-hPTH (1-84). To improve the purification efficiency, thermoosmotic shock conditions were optimized and achieved as follows: the optimized high osmotic solution containing 20mM Tris-HCl buffer (pH 8.0), 1mM EDTA, and 25% sucrose; the low osmotic solution containing 20mM Tris-HCl buffer (pH 8.0), 1mM EDTA, and the heat-treatment temperature of 100 degrees C for 10 min. Using this method, the purity of Trx-hPTH (1-84) was up to 73% and the yield was up to 72%, respectively. In addition, the two separation methods of both thermoosmotic shock and affinity chromatography have been compared, indicating that thermoosmotic shock is an economical and feasible way for the fusion protein separation. Besides, the thermoosmotic shock method may be used for the purification of some proteins of thermal stability without N-terminal His-tag.

A pET system encoding the fusion protein gene of thioredoxin (Trx) and human parathyroid hormone (hPTH) was introduced into Escherichia coli BL21 (DE3). Recombinant Trx-hPTH fusion protein was expressed in soluble form in the cytoplasm of the E. coli transformant. To recover Trx-hPTH from the E. coli culture efficiently, a novel tactic was developed by adding Triton X-100 into the fermentation culture at the exponential growth phase of E. coli and by heat treatment of the culture at the end of the fermentation. A concentration of 1% (v/v) Triton X-100 was added into the culture at the same time as IPTG addition after optimization. Under these conditions, addition of Triton X-100 had little effect on the cell growth, but more than 75% of the total recombinant Trx-hPTH was released into the fermentation broth. Also, a much higher volumetric yield of recombinant Trx-hPTH could be obtained with protein release compared to yield without protein release. Simultaneously, owing to the highly thermal stability of Trx-hPTH fusion protein, heat treatment of the fermentation broth at 80 degrees C for 15 min at the end of fermentation was employed for primary purification. Results demonstrated that heat treatment not only boosted further release of the recombinant Trx-hPTH fusion protein into the fermentation broth but also precipitated/denatured most of the nontarget proteins released in the broth. The tactics described herein integrated the fermentation process with subsequent recovery steps and thus provided a valuable and economical method for the production of Trx-hPTH and maybe some other Trx fusions in E. coli.