Engineering Escherichia coli for constitutive production of monophosphoryl lipid A vaccine adjuvant.

During the COVID-19 pandemic, expedient vaccine production has been slowed by the shortage of safe and effective raw materials, such as adjuvants, essential components to enhance the efficacy of vaccines. Monophosphoryl lipid A (MPLA) is a potent and safe adjuvant used in human vaccines, including the Shingles vaccine, Shingrix. 3-O-desacyl-4'-monophosphoryl lipid A (MPL), a representative MPLA adjuvant commercialized by GSK, was prepared via chemical conversion of precursors isolated from Salmonella typhimurium R595. However, the high price of these materials limits their use in premium vaccines. To combat the scarcity and high cost of safe raw materials for vaccines, we need to develop a feasible MPLA production method that is easily scaled up to meet industrial requirements. In this study, we engineered peptidoglycan and outer membrane biosynthetic pathways in Escherichia coli and developed a Escherichia coli strain, KHSC0055, that constitutively produces EcML (E. coli-produced monophosphoryl lipid A) without additives such as antibiotics or overexpression inducers. EcML production was optimized on an industrial scale via high-density fed-batch fermentation, and obtained 2.7 g of EcML (about 135,000 doses of vaccine) from a 30-L-scale fermentation. Using KHSC0055, we simplified the production process and decreased the production costs of MPLA. Then, we applied EcML purified from KHSC0055 as an adjuvant for a COVID-19 vaccine candidate (EuCorVac-19) currently in clinical trial stage III in the Philippines. By probing the efficacy and safety of EcML in humans, we established KHSC0055 as an efficient cell factory for MPLA adjuvant production.

Quantitative real-time PCR detection and analysis of a lumpy skin disease outbreak in Inner Mongolia Autonomous Region, China.

Lumpy skin disease (LSD) is a severe disease of bovine characterized by nodules on the skin, mucous membranes, and profuse nasal discharge which causes severe economic losses. In October 2020, an LSD outbreak case was found in Inner Mongolia Autonomous Region, China. A total of 1,206 cattle were sold from the same imported animal quarantine field to 36 farms after the quarantine period finished, and over 30 farmers reported symptoms such as skin scabs found in newly arrived cattle shortly after that. A large-scale LSD outbreak investigation was launched after laboratory diagnosis confirmed LSD. The clinical samples of 1,206 cattle from 36 farms, including 1,206 whole blood, 1,206 oral and nose swabs, and 355 scabs, were collected for the qRT-PCR test. The result showed that 51 whole blood samples (4.23%), 580 swab samples (48.09%), and 350 skin scabs (98.59%) were lumpy skin disease virus (LSDV) positive, 33 of 36 farms were affected. This study aims to provide a basis for LSD epidemiological traceability, movement control, and measures for prevention and control.

The effectiveness of e-healthcare interventions for mental health of nurses: A PRISMA-compliant systematic review of randomized controlled trials.

BACKGROUND: Mental health problems, including burnout among nurses, are common and important. With the rapid development of information and communication technologies and the rise in use of smartphones, the use of e-mental health strategies is increasing in public and clinical settings, and initial clinical trials using this intervention have been conducted. This systematic review evaluated whether e-healthcare interventions improve burnout and other mental health aspects in nurses. METHODS: Six electronic databases including MEDLINE (via PubMed), EMBASE (via Elsevier), the Cochrane Library Central Register of Controlled Trials, the Cumulative Index of Nursing and Allied Health Literature, the Allied and Complementary Medicine Database, and PsycARTICLES were searched to collect relevant randomized controlled trials up to January 28, 2021, using e-healthcare interventions for mental health in nurses. The e-healthcare intervention was classified as web-based, smartphone-based, and real-time online interventions. The primary outcome was burnout in this population. Due to the heterogeneity of the interventions used in the included studies, quantitative synthesis was not performed, but included studies were analyzed qualitatively. Also, the details of e-healthcare for the mental health of nurses were analyzed. The methodological quality of included studies was assessed using Cochrane's Risk of Bias tool. RESULTS: Seven randomized controlled trials were included in this study. The 20-minute session of an online form of the emotional freedom technique was reported to significantly improve burnout severity compared to no intervention (P < .001). Other outcomes, such as career identity, quality of work life, workplace bullying, job stress, turnover intention, distress, anxiety, and resilience in nurses, were also reported to be improved by e-healthcare interventions. The methodological quality of the included studies was generally poor. CONCLUSIONS: In conclusion, there was some evidence that e-healthcare interventions may improve mental health outcomes, including burnout in nurses, compared with no intervention. However, due to the poor methodological quality and wide heterogeneity of the interventions and outcomes in the included studies, we were not able to reach sufficiently reliable conclusions. E-healthcare intervention for nurses in the new coronavirus disease era was discussed. High-quality clinical trials in this area should be conducted in the future.

Polarization of human gingival fibroblasts by Th1-, Th2-, Th17-, and Treg-derived cytokines.

BACKGROUND AND OBJECTIVE: The purpose of this study was to evaluate whether gingival fibroblasts (GFs) can be differently activated and polarized into distinct functional subtypes by T-helper (Th) cytokines. METHODS: Gingival fibroblasts were stimulated with interferon (IFN)-γ, interleukin (IL)-4, IL-17, and transforming growth factor (TGF)-ß, representative cytokines of Th1, Th2, Th17, and regulatory T cells, respectively, and the gene expression profiles were analyzed by microarray. Differentially expressed genes (DEGs) in GFs stimulated by 4 cytokines were screened, and a gene ontology (GO) analysis of the DEGs was conducted. To confirm the reliability of the microarray results, the DEGs that showed the largest differences compared with non-stimulated GFs were further analyzed by RT-PCR. To evaluate the effect of polarization on GFs responses to lipopolysaccharide (LPS), GFs stimulated by 4 cytokines were further stimulated with Escherichia coli LPS and mRNA levels of several genes were analyzed using RT-PCR. RESULTS: Differentially expressed genes by 4 Th cytokines were enriched in different GO terms, and the patterns of gene expression on GFs were shown functionally different. GFs stimulated with IFN-γ (GF(IFN-γ)) up-regulated the expression of chemokines (chemokine (C-X-C motif) ligand (CXCL)9, -10, -11, chemokine (C-C motif) ligand (CCL)8), molecules involved in antigen presentation, complement component 3 (C3), and other immune response-related molecules, whereas they down-regulated the expression of several types of collagen, extracellular matrix (ECM) components, and DNA replication and nuclear protein-related molecules. By contrast, GF(IL-4) up-regulated the expression of ECM components, cell adhesion molecules, and tissue development-related molecules and down-regulated the expression of chemokines (CXCL10 and CXCL8) and adaptive immune response-related molecules. GF(IL-17) up-regulated the expression of chemokines and other molecules for neutrophil infiltration and activation, the pro-inflammatory cytokine IL-6, and C3. GF(TGF-ß) up-regulated the expression of cell growth-related molecules, ECM components, several types of collagen, and cell adhesion molecules and down-regulated the expression of molecules related to complement activation and bacterial recognition. GFs stimulated by 4 cytokines responded differently to LPS. CONCLUSION: These results show that Th cytokines can polarize GFs into cells with functionally distinct features: immune-activating but tissue-destructive GF(IFN-γ), tissue-reparative, and immune-inhibiting GF(IL-4), highly pro-inflammatory GF(IL-17), and potent tissue-reparative GF(TGF-ß).

Preclinical Assessment of Paclitaxel- and Trastuzumab-Delivering Magnetic Nanoparticles Fe3O4 for Treatment and Imaging of HER2-Positive Breast Cancer.

Objective: The purpose of this study was to investigate the anticancer activity and the potential imaging use of the innovative combination of magnetic nanoparticles (MNPs)-Fe3O4, paclitaxel (PTX), and trastuzumab (Herceptin) in HER2-positive breast cancer. Methods: MNPs-Fe3O4 was synthesized and underwent water phase transfer and hydrophobic molecular loading, and its surface was then coupled with Herceptin mono-antibody. The morphological characteristics of MNPs-Fe3O4 were observed under transmission electron microscopy (TEM). Effects of PTX-Herceptin-MNPs-Fe3O4 on breast cancer cells were evaluated using the 3-(4,5-dimethylthiazol-2-yl)-2,4-diphenyltetrazolium bromide assay and the flow cytometric apoptosis assay. To establish a xenograft model, we injected breast cancer SK-BR-3 cells into the left thighs of nude mice. We measured the effect of PTX-Herceptin-MNPs-Fe3O4 on tumor growth by measuring tumor size and calculating inhibition rate with immunohistochemistry analysis further performed, and analyzed MNPs-Fe3O4 accumulation in tumor lesions using in vivo magnetic resonance imaging and in vivo fluorescence imaging. Results: Most MNPs were in spherical shape of about 10 nm in diameter observed under TEM. PTX-Herceptin-MNPs-Fe3O4 showed greater cytotoxic effects, and induced a higher apoptosis rate of SK-BR-3 cells than all the other groups, with corresponding changes of apoptosis-related proteins. Meanwhile, the in vivo tumor xenograft model showed that tumor inhibition rate in the PTX-Herceptin-MNPs-Fe3O4 group was higher than in the PTX-Herceptin group. Furthermore, PTX-Herceptin-MNPs-Fe3O4 enhanced the T2 imaging contrast enhancement effect on tumors in tumor-bearing mice. Conclusion: The novel PTX-Herceptin-MNPs-Fe3O4 combination may represent a promising alternative breast cancer treatment strategy and may facilitate tumor imaging.

Effectiveness and safety of acupuncture in treating sleep disturbance in dementia patients: A PRISMA-compliant systematic review and limitations of current evidence.

BACKGROUND: Dementia is of increasing importance, as it is a major public health problem worldwide. Sleep disturbance is common in dementia patients and may be associated with worse cognitive symptoms or behavioral and psychological symptoms of dementia. Non-pharmacological approaches, such as acupuncture, for treating this clinical condition are gaining importance. This study aimed to comprehensively search and analyze randomized controlled clinical trials (RCTs) of acupuncture in treating sleep disturbance or sleep disorders in dementia patients. METHODS: A comprehensive search was conducted from 12 electronic databases on December 2, 2020. We included RCTs reporting the effectiveness and safety of acupuncture in treating sleep disorders or disturbance in dementia patients. The methodological quality of the included studies was assessed using the Cochrane Collaboration's risk-of-bias tool. RESULTS: Five articles with four original RCTs met the inclusion criteria. These studies reported clinical data suggesting that adjuvant acupuncture for hypnotics, and ear acupressure in dementia patients with sleep disorders or sleep disturbance may have clinical benefits in certain sleep-related parameters and total effective rate (TER). Only 1 study reported the safety profile of the intervention, and no acupuncture-related adverse reactions were reported. Some studies compared 2 kinds of acupuncture methods, and found that specific acupuncture methods were superior to conventional acupuncture in improving sleep-related parameters, cognitive function and TER. The methodological quality of the included clinical studies was not high. CONCLUSIONS: There were limited acupuncture studies on this topic. Given the number of studies included and their sample size, methodological quality, and heterogeneities, clinically relevant conclusions could not be drawn. Further clinical studies are needed in this field considering its urgency and importance.

The Effectiveness and Safety of Mind-Body Modalities for Mental Health of Nurses in Hospital Setting: A Systematic Review.

The mental health of nurses including burnout is an important issue. The purpose of this systematic review was to evaluate whether mind-body modalities improve burnout and other mental health aspects of nurses. A comprehensive search was conducted using six electronic databases. Randomized controlled trials using mind-body modalities on the mental health of nurses, up to January 2021, were included. The methodological quality of the included studies was assessed using the Cochrane Risk of Bias tool. Seventeen studies were included in the review. Data on mindfulness-based interventions (MBIs) and yoga were available for burnout, and there was no evidence that multimodal resilience programs including MBIs statistically significantly improved burnout levels compared to no intervention or active control groups. However, one study reported that yoga could significantly improve emotional exhaustion and depersonalization, which are subscales of burnout, compared to usual care. In addition, the effects of MBIs, relaxation, yoga, and music on various mental health outcomes and stress-related symptoms have been reported. In conclusion, there was some evidence that yoga was helpful for improvement in burnout of nurses. However, due to the heterogeneity of interventions and outcomes of the studies included, further high-quality clinical trials are needed on this topic in the future.

The efficacy evaluation of partial nephrectomy with selective renal artery branch occlusion by laparoscopy.

BACKGROUND: To investigate the clinical application and effect of laparoscopic partial nephrectomy with renal artery branch occlusion in the treatment of early renal tumors. METHODS: A retrospective analysis was conducted on the clinical data of 15 cases of renal tumor patients who underwent partial nephrectomy by laparoscopic selective renal artery branch occlusion in our department from January 2017 to January 2018. Nine male patients and 6 female patients were aged 46 to 65 years, with an average age of 54.3 ±â€Š7.2 years. The diameters of tumors were 2.2 to 4.0 cm, with an average of 3.3 ±â€Š0.7 cm. There are 10 tumors locating on the left side and 5 on the right side. Preoperative renal glomerular filtration rate (GFR) were 77.3 to 61.9 mL/min with an average of 47.6 ±â€Š7.5 mL/min. All patients' diseased kidneys underwent renal computer tomography angiography examination before surgery. And the diseased kidney underwent reexamination of renal GFR. The operation time, renal artery branch occlusion time, intraoperative blood loss, postoperative hospital stay, changes of renal function, and complications were evaluated. RESULTS: All surgery were completed successfully, the surgery time was 136.7 ±â€Š15.2 min, intraoperative renal artery branch occlusion time was 21.3 ±â€Š4.5 min, the intraoperative blood loss was 223.3 ±â€Š69.5 mL, the postoperative hospital stay was 6.5 ±â€Š1.7 days, and the postoperative 1-month GFR was 49.5 ±â€Š6.6 mL/min. There was no significant difference between the renal GFR before and after surgery (P > .05). There was no blood transfusion and transfer open surgery cases. The patients were followed up for 3 to 15 months without complications. CONCLUSIONS: Partial nephrectomy with selective renal artery branch occlusion by laparoscopy is a safe, feasible, and effective method for the treatment of early renal cancer. It makes good use of the technical advantages of clear operation field and fine operation of laparoscopic surgery, avoids the heat ischemia process of the whole kidney, and can better protect the renal function.

Herbal medicine for behavioral and psychological symptoms of dementia: A protocol for systematic review.

BACKGROUND: Dementia is becoming a major public health problem worldwide with the aging of the world's population. Behavioral and psychological symptoms of dementia (BPSD), associated symptoms of dementia, not only predicts the poor prognosis of patients with dementia, but is also a major factor causing the care burden on caregivers, especially informal caregivers. For BPSD management, an alternative to existing psychotropic drugs is needed, given the benefit-harm ratio. Therefore, in this systematic review, we will evaluate the effectiveness and safety of herbal medicine for BPSD. METHODS AND ANALYSIS: Thirteen electronic databases will be comprehensively searched. Clinical studies reporting the efficacy (or effectiveness) and safety of herbal medicines in BSPD management published from their inception to December 2020 will be included. The primary outcome will be BPSD symptoms assessed by the validated tool. Moreover, total effective rate, daily living activities and quality of life of patients, burden and quality of life of caregiver, placement in a long-term care facility from home, and safety data will be regarded as the secondary outcome. Two independent researchers will perform the study selection, data extraction, and quality assessment process. To assess the methodological quality of the included studies, validated tools according to its design, such as the Cochrane Collaboration's risk of bias tool will be used. To perform meta-analysis, RevMan version 5.3 will be used, with mean differences for continuous outcomes and risk ratio for binary outcomes, and 95% confidence intervals. According to the heterogeneity and number of included studies, a fixed- or random-effects model will be used. REGISTRATION NUMBER: OSF (URL: https://osf.io/3u8ch), PROSPERO (CRD42020211000) (URL: https://www.crd.york.ac.uk/prospero/display_record.php?ID=CRD42020211000).

E. coli-Produced Monophosphoryl Lipid a Significantly Enhances Protective Immunity of Pandemic H1N1 Vaccine.

Emerging influenza viruses pose an extreme global risk to human health, resulting in an urgent need for effective vaccination against influenza infection. Adjuvants are vital components that can improve vaccine efficacy, yet only a few adjuvants have been licensed in human vaccines. Here, we investigate the adjuvant effects of Escherichia coli-produced monophosphoryl lipid A (MPL), named EcML, in enhancing the immunogenicity and efficacy of an influenza vaccine. Similar to MPL, EcML activated dendritic cells and enhanced the antigen processing of cells in vitro. Using ovalbumin (OVA) as a model antigen, EcML increased OVA-specific antibody production, cytotoxic T lymphocyte activity. The safety of EcML was demonstrated as being similar to that of MPL by showing not significant in vitro cell cytotoxicity but transient systemic inflammatory responses within 24 h in OVA immunized mice. Importantly, mice vaccinated with pandemic H1N1 (pH1N1) vaccine antigen, combined with EcML, were fully protected from pH1N1 virus infection by enhanced influenza-specific antibody titers, hemagglutination inhibition titers, and IFN-γ- secreting cells. Taken together, our results strongly suggest that EcML might be a promising vaccine adjuvant for preventing influenza virus infection.

Diagnostic performance of convolutional neural network-based Tanner-Whitehouse 3 bone age assessment system.

BACKGROUND: Bone age can reflect the true growth and development status of a child; thus, it plays a critical role in evaluating growth and endocrine disorders. This study established and validated an optimized Tanner-Whitehouse 3 artificial intelligence (TW3-AI) bone age assessment (BAA) system based on a convolutional neural network (CNN). METHODS: A data set of 9,059 clinical radiographs of the left hand was obtained from the picture archives and communication systems (PACS) between January 2012 and December 2016. Among these, 8,005/9,059 (88%) samples were treated as the training set for model implementation, 804/9,059 (9%) samples as the validation set for parameters optimization, and the remaining 250/9,059 (3%) samples were used to verify the accuracy and reliability of the model compared to that of 4 experienced endocrinologists and 2 experienced radiologists. The overall variation of TW3-metacarpophalangeal, radius, ulna and short bones (RUS) and TW3-Carpal bone score, as well as each bone (13 RUS + 7 Carpal) between reviewers and the AI, were compared by Bland-Altman (BA) chart and Kappa test, respectively. Furthermore, the time consumption between the model and reviewers was also compared. RESULTS: The performance of TW3-AI model was highly consistent with the reviewers' overall estimation, and the root mean square (RMS) was 0.50 years. The accuracy of the BAA of the TW3-AI model was better than the estimate of the reviewers. Further analysis revealed that human interpretations of the male capitate, hamate, the first distal and fifth middle phalanx and female capitate, the trapezoid, and the third and fifth middle phalanx, were most inconsistent. The average image processing time was 1.5±0.2 s in the TW3-AI model, which was significantly shorter than manual interpretation. CONCLUSIONS: The diagnostic performance of CNN-based TW3 BAA was accurate and timesaving, and possesses better stability compared to diagnostics made by experienced experts.

Metabolic engineering of Escherichia coli to produce a monophosphoryl lipid A adjuvant.

Monophosphoryl lipid A (MPLA) species, including MPL (a trade name of GlaxoSmithKline) and GLA (a trade name of Immune Design, a subsidiary of Merck), are widely used as an adjuvant in vaccines, allergy drugs, and immunotherapy to boost the immune response. Even though MPLA is a derivative of lipopolysaccharide (LPS), a component of the outer membrane of Gram-negative bacteria, bacterial strains producing MPLA have not been found in nature nor engineered. In fact, MPLA generation involves expensive and laborious procedures based on synthetic routes or chemical transformation of precursors isolated from Gram-negative bacteria. Here, we report the engineering of an Escherichia coli strain for in situ production and accumulation of MPLA. Furthermore, we establish a succinct method for purifying MPLA from the engineered E. coli strain. We show that the purified MPLA (named EcML) stimulates the mouse immune system to generate antigen-specific IgG antibodies similarly to commercially available MPLA, but with a dramatically reduced manufacturing time and cost. Our system, employing the first engineered E. coli strain that directly produces the adjuvant EcML, could transform the current standard of industrial MPLA production.

Changes in sICAM-1 and GM-CSF levels in skin tissue fluid and expression of IL-6, IL-17 and TNF-α in blood of patients with vitiligo.

Changes in the levels of soluble intercellular adhesion molecule-1 (sICAM-1) and granulocyte-macrophage colony stimulating factor (GM-CSF) in the skin tissue fluid, and the expression of interleukin (IL)-6, IL-17 and tumor necrosis factor-α (TNF-α) in the blood of patients with vitiligo were investigated. One hundred and twenty patients diagnosed with vitiligo and treated in Daqing Long Nan Hospital from March 2014 to March 2016 were selected, including 88 patients with vitiligo vulgaris and 32 patients with segmental vitiligo. Comparative analyses were performed for research indexes. Another 80 healthy volunteers receiving physical examination were selected as healthy controls. The levels of GM-CSF in tissue fluid were detected via radioimmunoassay (RIA). The levels of sICAM-1 in tissue fluid and IL-6, IL-17 and TNF-α in the blood were detected via enzyme-linked immunosorbent assay (ELISA). The expression levels of IL-6, IL-17 and TNF-α in patients with progressive vitiligo were significantly higher than those in patients with stable vitiligo (P<0.05). The levels of sICAM-1 and GM-CSF in the skin tissue fluid at white spots of patients with vitiligo vulgaris were significantly higher than those in the skin tissue fluid at non-white spots (P<0.05). sICAM-1 levels had significant positive correlations with the levels of IL-6, IL-17 and TNF-α in the blood (P<0.05). The levels of sICAM-1 in the skin tissue fluid and IL-6 in the blood of patients with vitiligo were negatively correlated with the course of disease (P<0.05). The levels of sICAM-1 in the skin tissue fluid and IL-6 and IL-17 in the blood of patients with vitiligo were positively correlated with the skin lesion area of patients (P<0.05). The levels of sICAM-1 and GM-CSF in the skin tissue fluid, and the expression levels of IL-6, IL-17 and TNF-α in the blood of patients with vitiligo are abnormal.

The Correlation Between Electrodiagnostic Results and Ultrasonographic Findings in the Severity of Carpal Tunnel Syndrome in Females.

OBJECTIVE: To determine which ultrasonographic measurement can be used as an indicator reflecting the severity of carpal tunnel syndrome (CTS), by comparing electrodiagnostic results with ultrasonographic measurements in females. Many previous studies have tried to reveal that the ultrasonography (US) can possibility be used for diagnosis and severity of CTS. However, the criteria are different by gender. Thus far, there have been many efforts towards providing patients with a CTS diagnosis and severity prediction using US, but studies' results are still unclear due to lack of data on gender differences. METHODS: We collected data from 54 female patients. We classified the severity of CTS according to electrodiagnostic results. Ultrasonographic measurements included proximal and distal cross-sectional areas of the median nerve and carpal tunnel. RESULTS: The severity by electrodiagnostic results statistically correlated to the proximal cross-sectional area (CSA) of the median nerve and carpal tunnel. However, there was no relationship between the proximal and distal nerve/tunnel indexes and the severity by electrodiagnostic results. CONCLUSION: In female patients with CTS, the proximal CSAs of the median nerve and carpal tunnel increase. They correlate with the severity by electrodiagnostic findings. The CSA of the proximal median nerve could be particularly used as a predictor of the severity of CTS in female patients. However, the nerve/tunnel index is constant, irrespective of the severity of CTS.

A novel microphysiometer based on high sensitivity LAPS and microfluidic system for cellular metabolism study and rapid drug screening.

This study presents a novel microphysiometer for studying the mechanism of cellular metabolism and drug effect. Based on the photocurrent amplification of light-addressable potentiometric sensor (LAPS), the constant voltage detection mode was introduced to enhance the detection sensitivity to replace the conventional constant current mode with the slow feedback rate. The photocurrent amplification of LAPS was improved by developing the sensor structure and fabrication processes. The sensor unit with microfluidic system was designed to detect the concentration change of cellular acidic metabolites in the extracellular microenvironment rapidly. Characteristic test experiments and cellular metabolism experiments were carried out to determine the performance of microphysiometer. The result showed that sensitivity of microphysiometer is significantly enhanced to sense the fluctuation of cellular metabolism rapidly and sensitively in real-time detection of living cells under physiological condition. With these improvements, the novel microphysiometer holds promise as a utility platform for studying cellular metabolism and evaluating drug effect.

Characterization and anti-allergic effect of a polysaccharide from the flower buds of Lonicera japonica.

A water-soluble polysaccharide (LJP-1), with a molecular weight of 1.8×10(5) Da, was isolated from the flower buds of Lonicera japonica. Gas chromatography (GC) analysis showed that the LJP-1 was mainly composed of d-glucose and a small amount of d-arabinose. On the basis of methylation analysis, LJP-1 had the backbone chain mainly consisting of 1,6-linked Glc and 1,3,6-linked Glc, which was terminated with 1-linked Ara residues at the O-3 position of 1,3,6-linked Glc in a relative molar ratio of 2.9:1:0.9. The anti-allergic effect of LJP-1 was evaluated on allergic contact dermatitis (ACD) induced by picryl chloride (PC) in mouse ear. Similar to prednisolone, orally administrated LJP-1 (20, 40 and 80 mg/kg) potently inhibited the PC-induced ACD, leading to substantial reductions in ear thickness, serum level of IgE and histamine, as well as tissue TNF-α. These results demonstrate that treatment with LJP-1 may be effective for preventing the development of PC-induced ACD.

Ectopic expression of soybean GmKNT1 in Arabidopsis results in altered leaf morphology and flower identity.

Plant morphology is specified by leaves and flowers, and the shoot apical meristem (SAM) defines the architecture of plant leaves and flowers. Here, we reported the characterization of a soybean KNOX gene GmKNT1, which was highly homologous to Arabidopsis STM. The GmKNT1 was strongly expressed in roots, flowers and developing seeds. Its expression could be induced by IAA, ABA and JA, but inhibited by GA or cytokinin. Staining of the transgenic plants overexpressing GmKNT1-GUS fusion protein revealed that the GmKNT1 was mainly expressed at lobe region, SAM of young leaves, sepal and carpel, not in seed and mature leaves. Scanning electron microscopy (SEM) disclosed multiple changes in morphology of the epidermal cells and stigma. The transgenic Arabidopsis plants overexpressing the GmKNT1 showed small and lobed leaves, shortened internodes and small clustered inflorescence. The lobed leaves might result from the function of the meristems located at the boundary of the leaf. Compared with wild type plants, transgenic plants had higher expression of the SAM-related genes including the CUP, WUS, CUC1, KNAT2 and KNAT6. These results indicated that the GmKNT1 could affect multiple aspects of plant growth and development by regulation of downstream genes expression.

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Synbiotic modulation of gut flora: effect on minimal hepatic encephalopathy in patients with cirrhosis.

Minimal hepatic encephalopathy (MHE) is an important disorder that may seriously impair daily functioning and quality of life in patients with cirrhosis. Treatment with lactulose is of benefit. The possible role of synbiotics (probiotics and fermentable fiber) has not been assessed. We screened 97 consecutive cirrhotic patients without overt hepatic encephalopathy for MHE using the number connection test and measurement of brainstem auditory evoked potentials. MHE, defined by abnormality on at least one test modality, was present in 58 (60%) patients. Fifty-five of these patients with MHE were randomized to receive a synbiotic preparation (n = 20), fermentable fiber alone (n = 20), or placebo (n = 15) for 30 days. Cirrhotic patients with MHE were found to have substantial derangements in the gut microecology, with significant fecal overgrowth of potentially pathogenic Escherichia coli and Staphylococcal species. Synbiotic treatment significantly increased the fecal content of non-urease-producing Lactobacillus species at the expense of these other bacterial species. Such modulation of the gut flora was associated with a significant reduction in blood ammonia levels and reversal of MHE in 50% of patients. Synbiotic treatment was also associated with a significant reduction in endotoxemia. The Child-Turcotte-Pugh functional class improved in nearly 50% of cases. Treatment with fermentable fiber alone was also of benefit in a substantial proportion of patients. In conclusion, treatment with synbiotics or fermentable fiber is an alternative to lactulose for the management of MHE in patients with cirrhosis.