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BACKGROUND: Local recurrence of colorectal cancer is associated with poor prognosis and quality of life. For patients not eligible for curative surgery, chemoradiation could be a promising therapeutic option, but there is no consensus yet for the concurrent chemotherapy regimen. This study evaluated the effects and safety of intensity-modulated radiation therapy (IMRT) when administered concurrently with raltitrexed and irinotecan to patients with unresectable recurrent colorectal cancer. METHODS: Eligible patients developed unresectable recurrent colorectal cancer, and were refractory to, or intolerant of, chemotherapy with fluoropyrimidine and oxaliplatin. IMRT was delivered (total dose: 50–60 Gy in 25–30 fractions) concurrently with irinotecan and raltitrexed (200 and 3 mg/m(2), respectively, on days 1 and 22). After treatment completion, patients underwent surgery or continued the same regimen of chemotherapy and were assessed by a multidisciplinary team. The primary endpoint was the objective response rate, defined as the proportion of patients with a confirmed complete response or partial response, assessed by radiologist and investigator after the completion of radiotherapy and reconfirmed a month later, in accordance with the Response Evaluation Criteria in Solid Tumors version 1.1. RESULTS: All 30 patients enrolled in this study between January 2019 and July 2020 completed radiotherapy and received a median of five chemotherapy cycles (range, 2–10 cycles). Twelve patients (40.0%) experienced an objective response (two complete responses and ten partial responses) and 17 patients exhibited stable disease [disease control rate (DCR): 96.7%]. The median follow-up was 22 months (range, 4–35 months), by the end of follow-up, six (20.0%) patients had local failure in the irradiation field, four (13.3%) had regional progression outside the irradiation field, 13 (43.3%) had distant metastasis or metastatic progression and nine (30.0%) died. The median progression-free survival (PFS) and local PFS (LPFS) were 13.5 and 23 months, respectively. The incidence of grade 3 or 4 adverse events was 26.7%, the most common of which was neutropenia (13.3%). CONCLUSIONS: IMRT with concurrent raltitrexed and irinotecan is a feasible treatment for unresectable recurrent colorectal cancer, which allows good tumor response and local control with acceptable toxicity profile.

Primordia formation is the first and most critical step in the development of fruiting bodies of edible fungi. In this study, the effects of exogenous ascorbic acid (ASA) on the Pleurotus ostreatus mycelia growth and primordia formation were researched and the results showed that the growth rate of P. ostreatus mycelia was accelerated and the time of primordia formation was advanced. The protein content and ascorbate oxidase (AAO) activity analysis showed that with the increase of ASA concentration, the protein content of mycelia first decreased and then increased, and in a certain concentration range, exogenous ASA could significantly promote the activity of AAO. Further expression analysis of the development regulating genes (Pofst3 and Pofst4) as well as blue light receptor coding genes (PoWC-1 and PoWC-2) showed the expression levels of those four genes all changed after the exogenous ASA addition, which indicated that the expression changes of PoWC-1 and PoWC-2, two key genes in the light morphogenesis, might affect the expression levels of development regulating genes Pofst3 and Pofst4, so as to lead to the formation of primordia in advance.

Objective Optimal approaches to patients with local recurrence of rectal cancer are unclear in China. This study aimed to evaluaty -30te the clinical outcomes and toxicity associated with different treatment regimens for patients with local recurrence of rectal cancer. Methods A retrospective chart review of patients with local recurrence of rectal cancer and previous radical surgical treatment between March 2010 and December 2017 with curative intent was performed. Disease-related endpoints included treatment progression-free survival (PFS) and overall survival (OS) using the Kaplan-Meier method. Toxicities were assessed using Common Terminology Criteria for Adverse Events, version 5.0, and complications were scored according to the Clavien-Dindo classification. Results A total of 71 patients met the inclusion criteria in this study. The recurrence sites were mainly local recurrence in the pelvic cavity and regional lymph node metastasis. Twenty patients received chemoradiotherapy combined with surgery, 10 underwent surgery alone, and others received chemoradiotherapy-alone (n = 27) and chemotherapy-alone (n = 14) treatment. A clear difference was found in PFS between surgery/chemoradiotherapy with surgery and chemoradiotherapy/chemotherapy groups (26.6 months vs 14.1 months, P = 0.033). The PFS of patients in the surgery combined with chemoradiotherapy, surgery alone, and chemotherapy/chemoradiotherapy groups was 65.2 months, 20.2 months, and 14.2 months, respectively (P = 0.042). The multivariate analysis of PFS demonstrated that surgery was an independent factor. The proportion of patients with distant metastases after chemoradiotherapy/chemotherapy was higher than that of patients undergoing surgery (36.6% vs 21.4%, P = 0.179). The OS of patients in the surgery combined with chemoradiotherapy, surgery alone, and chemotherapy/chemoradiotherapy groups was 89.4 months, 66.0 months, and 62.8 months, respectively (P = 0.189). Radiation treatment and surgery did not increase extra severe toxicities. Conclusion Surgery combined with chemoradiotherapy was a beneficial treatment mode for managing patients with locally recurrent, nonmetastatic rectal cancer. It was associated with better local disease control, no increase in toxicity, and prolonged survival among patients with locally recurrent rectal cancer.

Epidermolysis bullosa (EB) is a heritable blistering disorder. We performed a next-generation sequencing-based multigene panel test and successfully predicted 100% of the EB types, including, 36 EB simplex (EBS), 13 junctional EB (JEB), 86 dystrophic EB (DEB), and 3 Kindler EB. Chinese JEB and recessive DEB (RDEB) patients have relatively mild phenotypes; for severe type separately accounts for 45.5% and 23.8%, respectively. We identified 96 novel and 49 recurrent pathogenic variants in 11 genes, although we failed to detect the second mutation in one JEB and five RDEB patients. We identified one novel p.E475K mosaic mutation in the clinically normal mother of one out of 13 EBS patients with KRT5 mutations, one recurrent p.G2034R mosaic mutation, and one novel p.G2043R mosaic mutation in the clinically normal relatives of two out of 19 dominant DEB patients. This study shows that next-generation technology could be an effective tool in diagnosing EB.

Three polysaccharides (WZP1, WZP2, WZP3) and their Se-enriched products (SeWZP1, SeWZP2 and SeWZP3) were obtained from Pleurotus ostreatus using a simple, rapid method and HNO3-Na2SeO3 method, respectively. The molecular weight distribution profiles of all samples except SeWZP2 showed double peaks. The average molecular weights (Mw) of WZP1–3 were 48.6 kDa, 20.2 kDa and 11.8 kDa, respectively, and of SeWZP1–3 were 19.6 kDa, 37.7 kDa, 14.5 kDa, respectively. The complexity of monosaccharide composition of WZP1–3 was inversely proportional to the ethanol concentration used in the ethanol precipitation process. Additionally, the results of biological activity tests indicated that α-glucosidase inhibitory activity of WZP1–3 was related to the molecular weight and the monosaccharide composition complexity. The selenized modification can improve the α-glucosidase-inhibiting, hydroxyl radical-scavenging activity of P. ostreatus polysaccharides. Therefore, by improving their bioactivities by selenization, the polysaccharides of P. ostreatus could be utilized as a natural health food supplement.

Background:Locally recurrent colorectal cancer is often associated with considerable morbidity and poor quality of life. Moreover, surgical resection is frequently not viable because of tumor fixation in the pelvis. This study aimed to evaluate the effects and safety of intensity-modulated radiotherapy when administered concurrently with raltitrexed and irinotecan to patients with unresectable recurrent colorectal cancer.Methods:Eligible patients had unresectable pelvic recurrence of colorectal cancer, UGT1A1 genotype *1*1 or *1*28, and were refractory to, or intolerant of, chemotherapy with fluoropyrimidine and oxaliplatin. Intensity-modulated radiation therapy (IMRT) was delivered to the pelvis with a total dose of 50–60 Gy in 25–30 fractions, concurrently with irinotecan (200 mg/m2 on days 1 and 22) and raltitrexed (3 mg/m2 on days 1 and 22). After completion of radiation treatment, patients underwent surgery or continued the same regimen of chemotherapy and were assessed by a multidisciplinary team. The primary endpoint was the objective response rate, evaluated using RECIST version 1.1. Secondary endpoints were disease control rate (DCR), progression-free survival (PFS), local progression-free survival (LPFS), and safety. Clinical and imaging evaluations were scheduled every month during treatment.Results:Between January 1, 2019, and July 14, 2020, 30 patients were enrolled in this study at the Fudan University Shanghai Cancer Center. All patients completed radiotherapy with a median number of 5 chemotherapy cycles (range, 2–10). Twelve patients (40%) experienced an objective response, including two complete responses and ten partial responses. Seventeen patients exhibited stable disease, leading to a DCR of 96.7%. With a median follow-up of 13 (range, 4–25) months, progression was observed in 20 patients (11 loco-regional failures, 11 distant metastases, and 5 deaths). The median PFS was 13 (95% confidence interval [CI], 9–18) months; the median LPFS was 15 (95% CI, 14 to not reached) months. The incidence of grade 3 or 4 adverse events was 26.7%. The most common grade 3 or 4 adverse event was neutropenia (13.3%).Conclusions:IMRT with concurrent raltitrexed and irinotecan exhibited encouraging efficacy with an acceptable toxicity profile in patients with unresectable recurrent colorectal cancer.Trial registration: ClinicalTrials.gov: NCT04499586, registered on July 31, 2020 - Retrospectively registered, https://clinicaltrials.gov/ct2/show/NCT04499586.

Background This study discusses the effectiveness of artificial intelligence in one-to-one psychological intervention system for middle school students. The purpose of this study is to determine whether middle school students have enough trust in AI machines, to explore their internal psychology, and to evaluate the intervention effect of AI in middle school students' one-to-one psychological intervention system. Subjects and Methods 76 eighth grade students from two parallel classes in the same grade were randomly assigned to the same teacher. The research tools used include two sets of autonomic nerve detectors produced by South Korean medicore company and two sets of self-developed artificial intelligence one-to-one psychological intervention systems, including two artificial intelligence robots Xiaoxin and two emotional bracelets. 36 students in the intervention group received one-on-one intervention of artificial intelligence robot Xiaoxin, and t-test was conducted on physical stress, psychological stress, stress index, sympathetic nerve activity, stress resistance and average heart rate. Test participants can effectively alleviate psychological stress and prevent depression. The data showed that the DASS-21 comprehensive score of subjects participating in the test was 1.7 times that of subjects not participating in the test. In contrast, without participation, they are more likely to show depression, anxiety and stress, and tend to be passive in simple coping styles. The improved method can effectively alleviate the psychological pressure of employees and improve the ability to actively deal with pressure. Results Except for the four indexes of nerve stress and sympathetic nerve activity, P < 0.001. After the intervention, the average heart rate, physical stress and stress index decreased significantly, while the anti stress ability increased significantly. Statistical analysis shows that the one-to-one intervention of artificial intelligence robot Xiaoxin can effectively alleviate students' physical stress and stress index, and improve students' stress resistance. The intermediary effect analysis of emotional arousal found that the direct effect of anger on MCT fluency was significant, B = 3.80, P < 0.001, CI = [1.99, 5.62], and the indirect effect of arousal on MCT fluency was significant, B = 0.58, CI = [0.08, 1.66]; The direct effect of anger on MCT anger was significant, B = 6.08, P < 0.001, CI = [3.26, 8.89], and the indirect effect of arousal on MCT anger was significant, B = 0.80, CI = [0.08, 2.41]; The direct effect of anger on MCT harmfulness was significant, B = 0.42, P = 0.004, CI = [0.14, 0.69], and the indirect effect of arousal on MCT harmfulness was not significant, B = 0.03, CI = [− 0.03, 0.18]. The direct effect of anger on aut fluency was significant, B = 2.58, P = 0.032, CI = [0.23, 4.92], and the indirect effect of arousal on aut fluency was significant, B = 1.22, CI = [0.27, 2.62]); The direct effect of anger on aut novelty was significant, B = 4.60, P = 0.008, CI = [1.23, 7.96], and the indirect effect of arousal on aut novelty was significant, B = 1.79, CI = [0.42, 3.91] (see Figure 4D). The above results show that emotional arousal plays a partial mediating role in the influence of anger on the fluency and novelty of malicious creativity and general creativity, but it has no mediating role in the influence of anger on the harmfulness of malicious creativity. Conclusion Artificial intelligence one-to-one psychological intervention system is effective. Among the 38 students in the intervention group, this shows that most students have a high sense of trust in the artificial intelligence robot Xiaoxin. This sense of trust is enough to make the artificial intelligence one-to-one psychological intervention system play the role of psychological intervention. We should fully understand the important position of artificial intelligence robot in emotional factors, actively improve its role psychological ability, pay special attention to its impact on life attitude and healthy behavior, and be a healthy self-manager, coordinator and collaborator. According to the psychological characteristics of these groups, we should actively and flexibly organize colorful activities, strive to create a relaxed and harmonious living atmosphere, create a friendly and mutually beneficial emotional artificial intelligence relationship, encourage and guide these groups, adjust their psychology to the best state, and promote the healthy development of their good cognition and life.

Background Survival benefit of adjuvant radiotherapy for locally advanced gastric cancer following gastrectomy plus D2 lymphadenectomy has always been controversial. Esophagogastric junction (EGJ) adenocarcinoma, which is usually classified as gastric cancer in East Asia, often has a higher locoregional recurrence rate after operation because of its special anatomical characteristics. The aim of this study is to determine whether adjuvant radiotherapy can improve survival of locally advanced EGJ adenocarcinoma after D2 radical resection. Methods In this phase III, randomized, open label, controlled trial, we plan to recruit 378 patients with Siewert type II and III adenocarcinoma of EGJ, who had undergone transabdominal radical surgery and D2 lymphadenectomy, and were divided into pathological stage IIB to IIIC. All patients will be randomized 1:1 to receive either adjuvant chemotherapy alone (control group) or adjuvant chemotherapy plus chemoradiotherapy (experimental group). Patients allocated to control group will receive eight cycles of S-1 plus oxaliplatin (SOX), while the experimental group will receive two cycles of SOX followed by 45-Gy RT combined with S-1 and four additional cycles of SOX. The primary endpoint is 3-year disease-free survival rate (DFS). The secondary endpoints are 3-year overall survival rate (OS), 3-year locoregional recurrence-free survival rate (LRFS), 3-year distant metastasis-free survival rate (DMFS), and quality of life (QoL). Discussion In the past, the adjuvant treatment of EGJ adenocarcinoma needs to draw on the experience of esophageal adenocarcinoma or gastric adenocarcinoma. In this study, EGJ adenocarcinoma is considered as an independent disease, and the conclusion will provide evidence for optimal adjuvant therapy of locally advanced EGJ adenocarcinoma after D2 radical resection. Trial registration ClinicalTrials.govNCT03973008. Registered on 1 June 2019 (retrospectively registered), URL: https://clinicaltrials.gov/ct2/show/NCT03973008?term=NCT03973008&draw=2&rank=1

Background Atopic dermatitis (AD) is the most common skin disorder in infancy. However, the diagnosis and definite significance of infantile AD remains a debated issue. Objective To analyse the phenotypes of AD in infancy, to establish diagnostic criteria and to estimate the prevalence of this condition in China. Methods This is a multicentric study, in which 12 locations were chosen from different metropolitan areas of China. Following careful and complete history-taking and skin examination, the definite diagnosis of AD was made and the severity based on the SCORAD index was determined by local experienced dermatologists. Based on the detailed phenotyping, the major and representative clinical features of infantile AD were selected to establish the diagnostic criteria and evaluate their diagnostic efficacy. Results A total of 5967 infants were included in this study. The overall point prevalence of AD was 30.48%. The infantile AD developed as early as at the second month of life, and its incidence peaked in the third month of life at 40.81%. The proportion of mild, moderate and severe AD was 67.40%, 30.57% and 2.03%, respectively. The most commonly seen manifestations in the infantile AD were facial dermatitis (72.07%), xerosis (42.72%) and scalp dermatitis (27.93%). We established the novel diagnostic criteria of infants, which included: (i) onset after 2 weeks of birth; (ii) pruritus and/or irritability and sleeplessness comparable with lesions; and (iii) all two items above with one of the following items can reach a diagnosis of AD: (i) eczematous lesions distributed on cheeks and/or scalp and/or extensor limbs, and (ii) eczematous lesions on any other parts of body accompanied by xerosis. Conclusions In China, the prevalence of AD in infancy is 30.48% according to clinical diagnosis of dermatologists. The novel Chinese diagnostic criteria for AD in infants show a higher sensitivity and comparable specificity.

Only a few transcription factors involved in mushroom development have been reported till now, Therefore, identification of transcription factors in common edible mushroom has commercial and scientific importance. In this study, the Pofst3 gene from Pleurotus ostreatus was cloned and characterized. Bioinformatics analysis showed that Pofst3 protein had 71% sequence similarity with fst3 of model mushroom Schizophyllum commune. Furthermore, the function of Pofst3 gene was analyzed by overexpression and antisense silencing in P. ostreatus via Agrobacterium-mediated transformation. Expression verification of Pofst3 in transformants through qRT-PCR showed that compared with the wild type strains, the transcription level was about 1.26 ∼ 9.59 and 0.01 ∼ 0.30 fold in Pofst3 overexpressing and silencing strains, respectively. Petri dish and bag cultivation tests of transgenic strains showed that the number of primordia and the type of fruiting bodies of Pofst3 overexpressing strains were consistent with the wild type strains, i.e. fewer primordia and larger fruiting bodies; the number of primordia formed by Pofst3 silencing strains were more than those of wild type strains, but fruiting bodies were smaller. It was very likely that Pofst3 was involved in the regulation of P. ostreatus development through inhibiting the formation of clusters of primordia.

As one of the epidermal nevus syndromes, Schimmelpenning-Feuerstein-Mims (SFM) is characterized by craniofacial nevus sebaceous (NS) and extracutaneous abnormalities (e.g., brain, eyes, and bone). Here, we report a case of a 4-year-old boy who presented with significant skin abnormalities (NS in the scalp, extensive epidermal nevus along Blaschko's lines), ocular abnormalities (strabismus), central nervous system abnormalities (seizure and mental retardation), lymphatic dysplasia (chylous pleural and pericardial effusion), cardiac abnormalities (patent foramen ovale), urogenital system abnormalities (cryptorchidism, hypospadias), and a tumor predisposition (embryonal rhabdomyosarcoma). DNA samples from NS, rhabdomyosarcoma, and peripheral blood leukocytes were analyzed by next-generation sequencing. A novel mutation in the HRAS gene (c.38G>T; p.Gly13Val) was detected in a mosaic state in NS, rhabdomyosarcoma, and peripheral blood leukocytes, with different ratio of heterozygous mutation (HRAS c.38G>T) of 39.90% (9412/23 588 reads), 73.03% (205 562/281 468 reads), and 14.16% (15 837/111 842 reads), respectively. By predicting the impact of the mutation on the biological function of protein, we found that the novel HRAS mutation (c.38G>T; p.Gly13Val) had the highest damaging scores among other HRAS mutations reported so far. This is the first reported SFM syndrome patient with novel mosaic HRAS mutation, which may help to expand the mutational spectrum of HRAS and better understand the role of HRAS in the disease.

Colorectal cancer (CRC) is one of the most common types of malignancy and the third most commonly diagnosed form of cancer worldwide, ranking as the fourth leading cause of cancer-associated mortality. MicroRNA (miR)-576-5p has been reported to be highly expressed in patients with CRC; however, its biological role remains unclear. The present study aimed therefore to investigate the biological role and underlying mechanism of miR-576-5p in CRC cell line SW480. The viability of SW480 cells following transfection with miR-576-5p mimic or inhibitor was analyzed using MTT assay. Wound healing and Transwell assays were performed to determine the cell migratory and invasive abilities, respectively. A dual luciferase reporter assay was used to verify the predicted binding site between miR-576-5p and Wnt5a. Reverse transcription-quantitative PCR and western blotting were used to analyze the expression levels of miR-576-5p, E-cadherin, N-cadherin, vimentin, Snail1, Wnt5a, β-catenin, c-myc, cyclin D1 and p/t-c-Jun. Using bioinformatics analysis, high expression of miR-576-5p was found not only in tumor tissues, compared with the normal tissue, but also in CRC cells, compared with NCM460 cells. Furthermore, the inhibition of miR-576-5p expression significantly decreased the cell viability and the migratory and invasive abilities of SW480 cells, and suppressed the epithelial-to-mesenchymal transition (EMT). In addition, miR-576-5p could interact with Wnt5a and regulate the expression level of Wnt5a in order to influence the activity of Wnt/β-catenin signaling. The results from rescue experiments further demonstrated that the effect of miR-576-5p overexpression on cell metastasis and EMT was reversed by Wnt5a overexpression or treatment with XAV-939, which is an inhibitor of the Wnt/β-catenin signaling pathway. In conclusion, the findings from the present study suggested that inhibition of miR-576-5p may suppress SW480 cell metastasis and EMT by targeting Wnt5a and regulating the Wnt5a-mediated Wnt/β-catenin signaling pathway, providing a potential therapeutic target for the treatment of CRC.

In the present study, the complete mitogenome of Clavaria fumosa, was sequenced, assembled, and compared. The complete mitogenome of C. fumosa is 256,807 bp in length and is the largest mitogenomes among all Basidiomycota mitogenomes reported. Comparative mitogenomic analysis indicated that the C. fumosa mitogenome contained the most introns and intronic ORFs among all fungal mitogenomes. Large intergenic regions, intronic regions, accumulation of repeat sequences and plasmid-derived genes together promoted the size expansion of the C. fumosa mitogenome. In addition, the rps3 gene was found subjected to positive selection between some Agaricales species. We found frequent intron gain/loss events in Agaricales mitogenomes, and four novel intron classes were detected in the C. fumosa mitogenome. Large-scale gene rearrangements were found occurred in Agaricales species and the C. fumosa mitogenome had a unique gene arrangement which differed from other Agaricales species. Phylogenetic analysis for 76 Basidiomycetes based on combined mitochondrial gene sets indicated that mitochondrial genes could be used as effective molecular markers for reconstructing evolution of Basidiomycota. The study served as the first report on the mitogenomes of the family Clavariaceae, which will promote the understanding of the genetics, evolution and taxonomy of C. fumosa and related species.

Cafe-au-lait macules (CALMs) affect the appearance of patients and can result in serious psychological problems. Successful treatments without adverse effects remain challenging. We designed a prospective, randomized, controlled, evaluator-blinded trial on 40 pediatric patients to compare the efficacy between a low-fluence 1064-nm Q-switched Nd:YAG laser and a Q-switched Nd:YAG 532-nm laser for the treatment of solitary CALMs in children. We randomly assigned participants into 2 groups. We treated those in the first group with 3 sessions of 532-nm QS laser at 1-month intervals, and those in the second group with 6 sessions of 1064-nm LFQS laser at 2-week intervals. We found no significant differences in treatment efficacy (p = 0.14). The 1064-nm laser group referred significantly less pain than the 532-nm laser group (p = 0.0001). Side effects were detected in 5 patients in the 532-nm laser group. The difference of the side effects was statistically significant (p = 0.04). Two patients in 532-nm laser group were recurred and none in 1064-nm laser group. On a univariate logistic regression analysis, lesions with brown color, small size, and irregular edges were significantly associated with better outcomes (> 50% clearance). Multivariate logistic regression analysis found that brown lesions and lesions with irregular edges had higher odds of getting > 50% clearance (p < 0.05). In conclusion, the 1064-nm LFQS laser produced fewer side effects, less pain, and shorter recovery time than the 532-nm laser. Irregular-bordered, smaller, brown lesions improved better than smooth-bordered, larger, light brown lesions. Moreover, the 1064-nm laser may be a better choice for treating large size CALMs. However, no significant differences were found in terms of the treatment efficacy and recurrence.

For the present work, a dual channel optical sensing composite for the detection of picric acid (PA) was constructed, using luminescent lanthanide MOF as supporting substrate and a rhodamine-based dye as sensing probe, respectively. This composite material was discussed in detail by means of electron microscopy images, XRD, IR, TGA, absorption and emission spectra. These rod-liked composite nanocrystals had dual sensing channels towards PA, including colorimetric sensing and ratiometric fluorescent sensing. A linear working curve with limit of detection as low as 10 μM was obtained. Good selectivity towards PA was confirmed. Their sensing mechanism was discussed and found as the combination of proton-triggered rhodamine structural transformation and nitrophenyl-triggered Eu emission quenching. This composite material showed a superior sensing performance over literature ones owing to its dual sensing channels and the possibility of naked eye detection.

The influence of different concentrations of fulvic acid at 0, 100, 200, and 400 mg/kg was evaluated during the course of composting with straw and mushroom residues as substrates. The optimal concentration of fulvic acid is 100 mg/Kg based on microbial characteristics, chemical parameters, and germination index testing. Nearly 80% of the microbial taxa responded significantly to fulvic acid over the composting period, with a dynamic change of the co-occurrence network from complex to simple and then to complex. Fulvic acid accelerated the progress of composting and reduced the emission of gases at the thermophilic phase. The optimal concentration of fulvic acid enriched the beneficial microorganisms Aeribacillus, Oceanobacillus, and Rhodospirillaceae, and decreased the abundances of pathogenic microorganisms Corynebacterium, Elizabethkingia, and Sarcocystidae. This study indicates a new strategy to optimize the composting process using the biostimulant fulvic acid.

Pleurotus ostreatus laccase gene poxc is transcriptionally induced by copper, and several putative MREs have been found and confirmed in its promoter region. However, the related transcription factors mediating copper response via MREs have not been reported. To isolate MRE binding proteins, we carried out yeast one-hybrid screens. Reporter genes containing two trimers of the cMRE2 and cMRE3 element were prepared and introduced into a yeast strain. The yeast was transformed with library cDNA that represents RNA isolated from CuSO4-treated fungi of P. ostreatus. From the screen of yeast transformants, we isolated ltf4 which encoded protein with HTH DNA binding domain. Electrophoretic mobility shift assay suggested direct and specific interaction of ltf4 with the MRE2 of poxc. Quantitative RT-PCR showed that the transcription of ltf4 was significantly up-regulated after the copper addition, and the expression trend was consistent with the poxc after copper introduction. The results indicated that ltf4 could interact with cMRE2 and thus participated in the regulation of copper mediated poxc transcription in P. ostreatus.

Cobalt and zinc bi-metal organic frameworks (CoZn-MOF) derived from double metal hydroxides (CoZn-DMH) is synthesized on the surface of rice husk carbon via hydrothermal method. The obtained CoZn-MOF/RHC is then used as electrochemical sensor for dopamine detection. SEM and TEM characterization displays that the morphology of CoZn-MOF/RHC is different from CoZn-DMH, as well as the electrochemical performance. Further investigation reveals that the kinetic process of the redox reaction of dopamine on CoZn-MOF/RHC electrode is controlled by diffusion process. Under the optimum conditions, the redox peak current linearly relates with the concentration of dopamine in the range of 2 μM~0.8 mM, with the low detection limit of 0.67 μM. During the anti-interference experiment, CoZn-MOF/RHC exhibits good selectivity toward dopamine detection but inappreciable response for interferent.

BACKGROUND Increasing evidence suggests that cancer-associated inflammation is associated with poorer outcomes. The neutrophil-to-lymphocyte ratio (NLR), considered as a systemic inflammation marker, is thought to predict prognoses in colorectal cancer. In this study, we explored the association between the NLR and prognoses following neoadjuvant chemoradiotherapy (CRT) for locally advanced rectal cancer (LARC). MATERIAL AND METHODS From February 2002 to December 2012, a group of 202 patients diagnosed with LARC and receiving neoadjuvant CRT followed by radical surgery was included in our retrospective study. The associations between the pre-CRT NLR and clinicopathological characteristics, as well as the predictive value of pre-CRT NLR against survival outcomes, were analyzed. RESULTS The average NLR was 2.7±1.5 (median 2.4, range 0.6-12.8). There were 63 (31.2%) patients with NLR ≥3.0, and 139 (68.8%) patients with NLR

This paper reported two rhodamine-derived chemosensors for cysteine optical sensing. To minimize their photobleaching, up-conversion NaYF 4 :Yb 3+ /Er 3+ nanocrystals were prepared and modified with α-cyclodextrin, serving as excitation host. Efficient energy transfer between this excitation host and our chemosensors was confirmed by spectral analysis, energy transfer radius analysis and emission decay lifetime comparison. Detailed analysis suggested that our chemosensors followed a simple recognition mechanism towards cysteine with binding stoichiometry of 1:1. Chemosensor emission was increased with increasing cysteine concentration. Good selectivity and linear sensing behavior were obtained. The correlation between sensing performance and chemosensor structure was tentatively discussed.

Geographic tongue (GT) is a benign inflammatory disorder of unknown etiology. Epidemiology and histopathology in previous studies found that generalized pustular psoriasis (GPP) is a factor associated with GT, but the molecular mechanism remains obscure. To investigate the mechanism of GT, with and without GPP, three cohorts were recruited to conduct genotyping of IL36RN, which is the causative gene of GPP. In a family spanning three generations and diagnosed with only GT (“GT alone”), GT was caused by the c.115+6T>C/p.Arg10ArgfsX1 mutation in the IL36RN gene. An autosomal dominant inheritance pattern with incomplete penetrance was observed. In the cohort consisting of sporadic cases of “GT alone” (n = 48), significant associations between GT and three IL36RN variants (c.115+6T>C/p.Arg10ArgfsX1, c.169G>A/p.Val57Ile and c.29G>A/p.Arg10Gln) were shown. In the GPP patient cohort (n = 56) and GPP family member cohort (n = 67), a significant association between the c.115+6T>C mutation and the simultaneous presence of GPP and GT was observed when compared to the presence of GPP without GT (P

Blue light is necessary for initiation of mushroom formation of Pleurotus ostreatus. In this study, we isolated homologues of the blue light receptor genes wc-1 and wc-2 from P. ostreatus, PoWC-1 and PoWC-2. The PoWC-1 contained three typical PAS domains and one PAS domain exhibited significant similarity to the LOV domain of known blue light receptors. The PoWC-2 had one typical PAS domain and one ZnF domain. The qRT-PCR analysis showed that PoWC-1 and PoWC-2 expression increased in a short time, and the final level tended to be stable along with the light illumination. The PoWC-1 and PoWC-2 expression levels of the primordium period was higher than that of mature fruiting-body period; and in the pileus were the highest, followed by the stipe and the gills. The expression of PoWC-1 and PoWC-2 in pre-primordial mycelia was induced by light exposure. In vivo analysis through yeast two-hybrid experiment disclosed that PoWC-1 and PoWC-2 could form heterologous complex to activate the reporter genes and the complex perform the transcription factor function requiring the addition of FAD.

Ethylene biosynthesis and function in Agaricus bisporus (the button mushroom) remain uncertain. The enzyme activities of 1-aminocyclopropane-1-carboxylate (ACC) synthase (ACS) and ACC oxidase (ACO) were detectable in A. bisporus AS2796 and inhibited by α-aminooxyacetic acid and Co2+. We cloned and sequenced 2 ACS genes (Ab-ACS1 and Ab-ACS2) and 1 ACO gene (Ab-ACO) from the mushroom strain. Ab-ACS1 and Ab-ACS2 demonstrated low amino acid sequence similarity. Ab-ACO demonstrated an amino acid sequence completely identical to that of ACO1_AGABI from A. bisporus. Antisense ACO significantly reduced ACO gene expression level, ACO enzyme activity, and ethylene production in the mushroom transformants. The transformants grew faster than the wild-type strain in sterilized compost and normally formed primordia when cultivated in sterilized compost with the sterilized casing vermiculite, but the wild-type strain did not. Our results show that ethylene is synthesized in button mushrooms via the ACC pathway. Ethylene inhibited button mushroom mycelial growth and development.

In order to isolate the differentially expressed genes in the primordium stage of Pleurotus ostreatus, the SSH cDNA library was constructed using the cDNA from dikaryotic mycelium stage as a driver and the cDNA from primordium stage as a tester. There were 423 significantly differently expressed clones among 2055 positive clones after three times of reverse Northern blot differential screening. After the repeated sequences being removed, 46 genes were identified which were putatively involved in cell rescue and defense, energy metabolism, transcription and protein regulation, membrane proteins, and signal transduction; 18 genes encoding hypothetical proteins with unknown function; 5 genes without any homology. PoALDH1 and its full-length cDNA sequence were cloned using the Aldehyde dehydrogenase EST isolated from the library. The amino acid sequence of PoALDH1 contains conservative glutamic acid and cysteine residues active sites of aldehyde dehydrogenase family. When exposed to different concentrations of sodium chloride, the mycelium growth was inhibited and the expression level of PoALDH1 was significantly higher than that of the control one, which indicated that PoALDH1 may have the ability to relieve salt stress. The results of this study will provide useful information for isolating growth and development related genes of P. ostreatus.

Recent evidences demonstrate that preoperative chemoradiotherapy (pCRT) followed by mesorectal excision is an effective therapy for patients with locally advanced rectal cancer (LARC). Nevertheless, the predictive molecular biomarkers for the response of patients to CRT remain largely unknown. Here we showed that the expression of miR-519b-3p was correlated with the responsiveness to pCRT in patients with LARC. We found that miR-519b-3p was highly expressed in responsive LARC samples. And we showed that miR-519b-3p may serve as a novel predictive marker by ROC analysis. In addition, overexpression of miR-519b-3p enhanced responsiveness to chemoradiation in vitro. Mechanistically, we found that miR-519b-3p directly bond to the 3' UTR of ARID4B mRNA whose expression was inversely correlated with miR-519b-3p expression. Finally, we performed functional experiments and showed that miR-519b-3p was directly involved in response to pCRT in rectal cancer patients in an ARID4B-dependent way.

The unusual amino acid l-cyclopropylalanine was isolated from the mushroom Amanita virgineoides after detection in an anti-fungal screening test. l-Cyclopropylalanine was found to exhibit broad-spectrum inhibition against fungi and bacteria. The anti-fungal activity was found to be abolished in the presence of the amino acid l-leucine, but not any other amino acids, indicating that l-cyclopropylalanine may block the biosynthesis of the essential amino acid l-leucine, thereby inhibiting fungal and bacteria growth. Further biochemical studies found l-cyclopropylalanine indeed inhibits α-isopropylmalate synthase (α-IMPS), the enzyme that catalyzes the rate-limiting step in the biosynthetic pathway of l-leucine. Inhibition of essential l-leucine synthesis in fungal and bacteria organisms, a pathway absent in host organisms such as humans, may represent a novel antibiotic mechanism to counter the ever-increasing problem of drug resistance to existing antibiotics.

ADDITIONAL INDEX WORDS. enzyme, respiration intensity, cultivation, Pleurotus ostreatus SUMMARY. We investigated a practical method for immobilizing liquid spawn of oyster mushroom (Pleurotus ostreatus) to prolong the storage time and provide convenient transportation of liquid spawn of edible mushrooms. The method was based on the mycelial pellets of liquid spawn adsorbed in carriers. Selected carriers were similar to cultivation substrates, and the best carrier was a mixture of cottonseed hull, corn core, and wheat bran with a ratio of 4.5:4.5:1 by weight. Immobilized spawn were prepared by mixing the pellets from liquid spawn with carriers using a ratio of 1:8 by weight. Within the first 15 days of storage at 20‐25 � C, the immobilized spawn grew strongly, respiration intensity and cellulase activitiesroserapidly,andthecountandbrightnessoftheisozymebandsofesterase, peroxidase, and polyphenol oxidase increased remarkably as well. From days 30 to 60, the cellulase activities fell and the brightness of the peroxidase and polyphenol oxidase bands gradually decreased, whereas the respiration intensity and the band count of esterase and peroxidase remained constant. After 60 days, the cultivated characteristics of the immobilized spawn were same as the fresh conventional solid cottonseed hull spawn. The results showed that immobilized spawn on the basis of the mycelial pellets of liquid spawn adsorbed in carrier can be used to extend the storage time and simplify transportation of liquid spawn of edible mushroom.

Two interesting cyathane-type diterpenoids, scabronines G and H, isolated from the fruiting bodies of the basidiomycete, Sarcodon scabrosus (L.: Fr.) Karst, were tested against the bacteria, Staphylococcus aureus, Bacillus thuringiensis, Escherichia coli, Bacillus megaterium and Bacillus subtilis. Both scabronine G and H showed promising activity in vitro against the five bacteria tested at 1 mg/ml and 100 μg/ml. The degree of antibacterial activity of both scabronine G and H was almost the same as streptomycin. Scabronines G and H were also tested against the pathogenic fungi, Gibberella zeae (Schwein.) Petch, Sclerotinia sclerotiorum (Lib.) de Bary, Fusarium moniliforme Sheldon, Fusarium oxysporum Schltdl., Fusarium oxysporum f. sp. vasinfectum, and Fusarium oxysporum f. sp. capsicum. Both scabronines G and H showed promising activity in vitro against G. zeae, S. sclerotiorum, F. moniliforme, F. oxysporum at 1 mg/ml and showed weak activity in vitro against F. oxysporum f. sp. vasinfectum and F. oxyspor...

This review surveys the chemical and biological literature dealing with the isolation, structural elucidation and bioactivity of hericenones and erinacines from the fruiting body and mycelium of Hericium erinaceus, concentrating on work that has appeared in the literature up to December 2009.

Locating sites of amino acids related to enzyme properties is still a challenge for rational engineering. Based on the strategy that sites of amino acids can be located by dipeptides, a computational model was made for pH-related dipeptides of xylanase. According to the dipeptide model, pH of a thermostable xylanase B from Thermotoga maritima was rationally designed by locating pH-related amino acids in its sequence and structure. In agreement with expectation, the optimum pH (pHopt) of the xylanase was improved by five amino acids substitutions: E70Q, E74Q, E77Q, G85Q and T87Q. In parallel assay reactions, at 90 °C, its pHopt increases to 5.5 from 5.1, and its whole pH profile also shifts 0.5 units towards alkaline area; at 80 °C, the relative activity decreases very little over a wide pH range from 5.25 to 6.0. This result demonstrated that the bioinformatics model is useful for pH rational design and engineering of xylanase, a model molecular of a large family of ∼10% proteins with (β/α)8-barrel structure.

This review surveys the chemical and biological literature dealing with the isolation, structure elucidation and bioactivity of diterpenoids from the fruiting bodies of macromycetes, concentrating on work that has appeared in the literature up to December 2007. In addition, this paper examines the research of diterpenoids produced by macromycetes grown in mycelial culture and the culture conditions for the fermentation of macromycetes. (© 2009 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim)

Dikarya is a subkingdom of fungi that includes Ascomycota and Basidiomycota. The gene expression patterns of dikaryon are poorly understood. In this study, we bred a dikaryon DK13 × 3 by mating monokaryons MK13 and MK3, which were from the basidiospores of Pleurotus ostreatus TD300. Using RNA-Seq, we obtained the transcriptomes of the three strains. We found that the total transcript numbers in the transcriptomes of the three strains were all more than ten thousand, and the expression profile in DK13 × 3 was more similar to MK13 than MK3. However, the genes involved in macromolecule utilization, cellular material synthesis, stress-resistance and signal transduction were much more up-regulated in the dikaryon than its constituent monokaryons. All possible modes of differential gene expression, when compared to constituent monokaryons, including the presence/absence variation, and additivity/nonadditivity gene expression in the dikaryon may contribute to heterosis. By sequencing the urease gene poure sequences and mRNA sequences, we identified the monoallelic expression of the poure gene in the dikaryon, and its transcript was from the parental monokaryon MK13. Furthermore, we discovered RNA editing in the poure gene mRNA of the three strains. These results suggest that the gene expression patterns in dikaryons should be similar to that of diploids during vegetative growth.

LEOPARD syndrome (LS) is an autosomal dominant inherited disorder primarily caused by mutations in the PTPN11, RAF1 and BRAF genes. Characteristic features include lentigines, craniofacial dysmorphism, myocardium or valve abnormalities, eletrocardiographic conduction defects and deafness. LS, neurofibromatosis type 1, Noonan syndrome and Legius syndrome are a group of highly overlapped disorders termed 'RASopathies'. Therefore, clinical discrimination between these syndromes represents a huge challenge. The present study reports a young child diagnosed with LS via identification of a common p.Thr468Met mutation in PTPN11. Taking into account two Taiwanese LS cases with an identical mutation, Thr468Met is likely to be the most prevalent mutation in the Chinese population. Furthermore, this study suggests that a clinical diagnosis of LS should be considered for individuals with congenital cardiac defects and atypical lentigines (i.e., light brown freckles) scattered particularly on the face.

The methanolic extract of the fruiting bodies of the mushroom Armillariella tabescens was found to show antifungal activity against Gibberella zeae. The active compound was isolated from the fruiting bodies of A. tabescens by bioassay-guided fractionation of the extract and identifi ed as armillarisin B. Armillarisin B eventually corresponds to 2-hydroxy-2- phenylpropanediamide and its structure was confi rmed on the basis of spectroscopic studies including 2D NMR experiments.

Mushroom β-glucans are potent immunological stimulators in medicine, but their productivities are very low. In this study, we successfully improved its production by promoter engineering in Pleurotus ostreatus. The promoter for β-1,3-glucan synthase gene (GLS) was replaced by the promoter of glyceraldehyde-3-phosphate dehydrogenase gene of Aspergillus nidulans. The homologous recombination fragment for swapping GLS promoter comprised five segments, which were fused by two rounds of combined touchdown PCR and overlap extension PCR (TD-OE PCR), and was introduced into P. ostreatus through PEG/CaCl2-mediated protoplast transformation. The transformants exhibited one to three fold higher transcription of GLS gene and produced 32% to 131% higher yield of β-glucans than the wild type. The polysaccharide yields had a significant positive correlation to the GLS gene expression. The infrared spectra of the polysaccharides all displayed the typical absorption peaks of β-glucans. This is the first report of successful swapping of promoters in filamentous fungi.

Three hygromycin B phosphotransferase (hph) gene expression systems for culinary-medicinal Oyster mushroom, Pleurotus ostreatus, plasmid pSHC, pAN7-1, and pBHt1 were evaluated through PEG/CaCl(2)-mediated protoplast transformation. Plasmid pSHC is a newly constructed hph gene expression system, composed of Escherichia coli hph gene, the P. ostreatus sdi promoter, and the CaMV35S terminator. The vector pAN7-1 was commonly used for integrative transformation in filamentous fungi. Plasmid pBHtl is a T-DNA binary vector, usually introduced into fungi by Agrobacterium-mediated transformation. The results showed that plasmids pSHC, pAN7-1, and pBHt1 were all integrated into the host chromosomes and expressed hygromycin B resistance in P. ostreatus. pAN7-1 had the highest transformation efficiency and hph gene expression level, pSHC the second, and pBHt1 the lowest. Growth rates of the transformants on plates containing hygromycin B were in correspondence with their hph gene expression levels. To our knowledge, this is the first report on integrated transformation of plasmid pAN7-1 and pBHt1 in P. ostreatus.

isolation and structure elucidation of two new aromatic compounds, hericenone I (I) and hericene D (II), with marked cytotoxic activity against EC109 cell line

isolation and structure elucidation of two new aromatic compounds, hericenone I (I) and hericene D (II), with marked cytotoxic activity against EC109 cell line

Many cultivated mushroom strains, such as Pleurotus ostreatus TD300, displayed symptoms of degeneration. A spherical virus POSV and four dsRNA segments were extracted from mycelium of P. ostreatus TD300. POSV had a diameter of 23 nm and encapsidated a 2.5 kb dsRNA segment with coat proteins whose molecular weights were 39 kDa and 30 kDa. Four dsRNA segments were 8.2 kb, 2.5 kb, 2.0 kb, and 1.1 kb in size, respectively. The 1.1 kb dsRNA segment often escaped detection. The cDNA and the amino acid sequences of the 8.2 kb dsRNA were homologous to those of RNA-dependent RNA polymerases (RDRP) of ssRNA oyster mushroom spherical virus (OMSV), and contained conserved motifs A to D which were almost identical to those in RDRP of OMSV. The cDNA and amino acid sequences of the 2.5 kb and 2.0 kb dsRNA segments were homologous to that of RDRP and capsid protein of dsRNA virus P. ostreatus virus 1 (PoV1), respectively. In particular, the amino acid sequence of 2.5 kb dsRNA segment had high identity with the conserved motifs A to C in RDRP of PoV1, a Partiviridae virus. After eliminating the viruses in P. ostreatus TD300, the symptoms of degeneration completely disappeared. The results reveal that P. ostreatus TD300 was at least infected by a particle virus POSV, and two naked viruses, one was a dsRNA virus with a 2.0 kb dsRNA segment, the other was an ssRNA virus whose replicating form of genome was an 8.2 kb dsRNA segment. Mycoviruses infection is a causative agent of mushroom strain degeneration.

Recently there is an increasing interest in changing the criterion of tumor classification from morphologic to molecular. In this perspective, the problem can be regarded as a classification problem in machine learning. In this study wavelet analysis is used to extract the features from high dimensional microarray profiles. To make it easier to find the significant genes, we remove the small change contained in the high frequency part based on wavelet decomposition. A set of orthogonal wavelet approximation coefficients is used to compress gene profiles and reduce the dimensionality. Experimental results show that approximation coefficients at 1st and 2nd level achieve good performance.