Diversity of Trichoderma species associated with soil in the Zoige alpine wetland of Southwest China.

The ecology of soil fungi is poorly understood, and recent comprehensive reports on Trichoderma are unavailable for any region, including the Zoige alpine wetland ecological region in China. One hundred soil samples were collected from different soil types and soil layers in Zoige alpine wetland ecological regions. Using the traditional suspension plating method, 80 Trichoderma strains were chosen to analyze species diversity. After a preliminary classification of morphological characteristics and the genes glyceraldehyde-3-phosphate dehydrogenase (gpd), 57 representative strains were selected and eventually identified as seven species via phylogenetic analyses of multilocus sequences based on the genes transcription elongation factor 1 alpha (tef1), encoding RNA polymerase II subunit B (rpb2) and ATP citrate lyase (acl1). Among them, T. harzianum was the dominant species isolated from five soil layers and four soil types, and had the highest isolation frequency (23%) in this zone, while T. polysporum and T. pyramidale were rare species, with isolation frequencies of less than 1%. Our detailed morphological observation and molecular phylogenetic analyses support the recognition of Trichoderma zoigense was described for the first time as a new species, while T. atrobrunneum as a new record for China was found. Our results will be used as a reference for a greater understanding of soil microbial resources, ecological rehabilitation and reconstructions in the Zoige alpine wetland.

A fatal outcome in a patient with coeliac disease who suspended a strict gluten-free diet: a case report.

Coeliac disease (CD) is an autoimmune small bowel disease that occurs in susceptible individuals that develop an immunological reaction to gluten. A strict gluten-free diet (GFD) is the primary treatment for CD. This case report describes a patient with CD recurrence due to a discontinuation of a strict GFD by the patient. After recurrence, the patient developed fever and pancytopaenia, and quickly died of haemophagocytic lymphohistiocytosis (HLH). To the best of our knowledge, this is the first description of a case of CD associated with HLH due to discontinued GFD, which may contribute to improving the awareness of the importance of maintaining a strict GFD and having regular follow-up examinations.

Pseudoglandular myxoid adrenocortical adenoma with positive epithelial markers: a case report and review of the literature.

Myxoid adrenal cortical adenoma with a pseudoglandular structure is a special histological variant and is extremely rare. We report about a 32-year-old Chinese woman with a right adrenal mass during a routine physical examination. The cut surface of the mass had a vague nodular, which gross appearance was pale, yellowish, and semitransparent. Histologically, the region is mostly characterised by pseudoglandular pattern with myxoid stroma. They are filled with clear cells or eosinophilic cells, as well as semitransparent regions, in which anastomosing small eosinophilic cells arranged in pseudoglandular, cord-like, or wreath-shaped structure float in the mucous pool. Immunohistochemical staining shows Melan-A, vimentin, and CD56 were positive and CK (AE1/AE3) were nucleus-side staining. A small number of tumor cells were positive for alpha-inhibin and synaptophysin, ki-67 labeling index was 3%. EMA, chromogranin A, WT-1, and P63 were negative. This report aimed to emphasize pseudoglandular patterns with mucus secretion which could occur in adenomas of the adrenal cortex, nucleus-side positive for CK is remarkable. However, this type may have malignant potential, so regular follow-up is needed.

Low-Dose Sirolimus Immunoregulation Therapy in Patients with Active Rheumatoid Arthritis: A 24-Week Follow-Up of the Randomized, Open-Label, Parallel-Controlled Trial.

BACKGROUND: We have reported previously the insufficient absolute number or functional defects of regulatory T cells (Tregs) in patients with rheumatoid arthritis (RA), challenging conventional unspecific immunosuppressive therapy. Sirolimus, a mTOR inhibitor, is reported to allow growth of functional Tregs; here, we investigated the efficacy of low-dose sirolimus combined with conventional immunosuppressants (sirolimus immunoregulation therapy) for RA treatment with lower side effects and better tolerance. METHODS: In this nonblinded and parallel-group trial, we randomly assigned 62 patients to receive conventional glucocorticoids and immunosuppressants with or without sirolimus at a dosage of 0.5 mg on alternate days for 24 weeks in a 2 : 1 ratio. The demographic features, clinical manifestations, and laboratory indicators including peripheral blood lymphocyte subgroups and CD4+T subsets were compared before and after the treatment. RESULTS: Finally, 37 patients in the sirolimus group and 18 in the conventional treated group completed the 6-month study. By 24 weeks, the patients with sirolimus experienced significant reduction in disease activity indicators including DAS28, ESR, and the number of tender joints and swollen joints (p < 0.001). Notably, they had a higher level of Tregs as compared with those with conventional therapy alone (p < 0.05), indicating that sirolimus could partly restore the reduced Tregs. Concomitantly, their usage of immunosuppressants for controlling disease activity was decreased as compared with the conventional group with no difference in blood routine, and liver and renal functions both before and after the treatment of sirolimus and between the two groups (p > 0.05). CONCLUSIONS: Low-dose sirolimus immunoregulatory therapy selectively upregulated Tregs and partly replaced the usage of immunosuppressants to control disease activity without overtreatment and evaluable side effect. Further study is required using a large sample of RA patients treated with sirolimus for a longer period. This trial is registered at the Chinese Clinical Trial Registry (http://www.chictr.org.cn/showproj.aspx?proj=17245).

Pathogenicity of an FAdV-4 isolate to chickens and its genomic analysis.

Fowl adenovirus serotype 4 (FAdV-4) strain SD1511 was isolated from chickens with severe inclusion body hepatitis and hydropericardium syndrome in Shandong Province, China. The isolate was cultured in primary chicken embryo kidney cells. A study of pathogenicity indicated that SD1511 readily infected 7-35-d-old chickens by intramuscular injection and intranasal and oral routes, causing 50%-100% mortality. The 35-d-old chickens suffered more severe infection than 7- and 21-d-old chickens with mortality highest in the intramuscular injection group. The serum from surviving chickens showed potent viral neutralizing capability. The complete genome of SD1511 was sequenced and analyzed. The strain was found to belong to the FAdV-4 cluster with more than 99% identity with the virulent FAdV-4 strains isolated in China in recent years except for some distinct variations, including deletions of open reading frame 27 (ORF27), ORF48, and part of ORF19. Our findings suggest that SD1511 might be used as a prototype strain for the study of pathogenesis and vaccine development.

Phenotypic and Genetic Characterization of Botrytis cinerea Population from Kiwifruit in Sichuan Province, China.

Botrytis cinerea (anamorph of Botryotinia fuckeliana) causes gray mold on numerous plants, including kiwifruit. The primary aim of this study was to investigate the phenotypic and genetic characteristics of the Botrytis cinerea population from kiwifruit in Sichuan Province, China. In all, 176 isolates were collected from kiwifruit orchards from eight geographic regions in Sichuan. All isolates were identified as B. cinerea sensu stricto based on the combined datasets, including morphological criteria, determination of the Bc-hch allele, and phylogenetic analysis of the genes RPB2, G3PDH, and HSP60. Three colony types (i.e., sclerotial, mycelial, and conidial) were observed on potato dextrose agar after 2 weeks, with sclerotial isolates, the predominant category, accounting for 40.91%. No obvious differences in microscopic characteristics were observed among the three types. Three genotypes of transposable elements were identified in the B. cinerea population: boty, flipper, and transposa types. The most prevalent genotype from different geographic populations of B. cinerea was transposa; in contrast, the flipper genotype accounted for only 3.98% of the total population, whereas the vacuma genotype was absent. According to MAT locus amplification, 87 and 89 isolates are MAT1-1 and MAT1-2 type, respectively, and the two mating types were found to be balanced overall in the population. Forty-eight representative isolates were all able to cause gray mold to some extent, and disease severities were significantly different between the cultivars Hongyang and Hort16A (P < 0.01). Disease severity was significantly greater on young leaves than on mature leaves (P < 0.01). No significant relationship was found between pathogenicity and geographical region, colony type, or transposon distribution. The results obtained in the present study suggest a relatively uniform species diversity of Botrytis but rich phenotypic and genetic differentiation within the B. cinerea population on kiwifruit in China. Utilizing resistant cultivars and rain-shelter cultivation instead of fungicides may be an effective approach to delaying pathogen variability.

Effects of oxygen transfer coefficient on dihydroxyacetone production from crude glycerol

Abstract The principal objective of this study was to evaluate the kinetics of dihydroxyacetone production by Gluconobacter frateurii CGMCC 5397 under different oxygen volumetric mass transfer coefficient (kLa) conditions in submerged bioreactors using biodiesel-derived crude glycerol as the carbon source. kLa is a key fermentation parameter for the production of dihydroxyacetone. Cultivations were conducted in baffled- and unbaffled-flask cultures (the kLa values were 24.32 h−1 and 52.05 h−1, respectively) and fed-batch cultures (the kLa values were held at 18.21 h−1, 46.03 h−1, and 82.14 h−1) to achieve high dihydroxyacetone concentration and productivity. The results showed that a high kLa could dramatically increase dihydroxyacetone concentrations and productivities. The baffled-flask culture (with a kLa of 52.05 h−1) favored glycerol utilization and dihydroxyacetone production, and a dihydroxyacetone concentration as high as 131.16 g/L was achieved. When the kLa was set to 82.14 h−1 in the fed-batch culture, the dihydroxyacetone concentration, productivity and yield were 175.44 g/L, 7.96 g/L/h and 0.89 g/g, respectively, all of which were significantly higher than those in previous studies and will benefit dihydroxyacetone industrial production.

Effects of oxygen transfer coefficient on dihydroxyacetone production from crude glycerol.

The principal objective of this study was to evaluate the kinetics of dihydroxyacetone production by Gluconobacter frateurii CGMCC 5397 under different oxygen volumetric mass transfer coefficient (kLa) conditions in submerged bioreactors using biodiesel-derived crude glycerol as the carbon source. kLa is a key fermentation parameter for the production of dihydroxyacetone. Cultivations were conducted in baffled- and unbaffled-flask cultures (the kLa values were 24.32h(-1) and 52.05h(-1), respectively) and fed-batch cultures (the kLa values were held at 18.21h(-1), 46.03h(-1), and 82.14h(-1)) to achieve high dihydroxyacetone concentration and productivity. The results showed that a high kLa could dramatically increase dihydroxyacetone concentrations and productivities. The baffled-flask culture (with a kLa of 52.05h(-1)) favored glycerol utilization and dihydroxyacetone production, and a dihydroxyacetone concentration as high as 131.16g/L was achieved. When the kLa was set to 82.14h(-1) in the fed-batch culture, the dihydroxyacetone concentration, productivity and yield were 175.44g/L, 7.96g/L/h and 0.89g/g, respectively, all of which were significantly higher than those in previous studies and will benefit dihydroxyacetone industrial production.

[Pregnancy outcome in 54 patients with antiphospholipid syndrome: a retrospective clinical study].

OBJECTIVE: To summarize the maternal/fetal outcome of pregnancy in antiphospholipid syndrome (APS) patients to evaluate the influence of treatment on the outcomes of pregnancy, and to investigate the possible clinical predictors of unsuccessful pregnancy. METHODS: The clinical characteristics, laboratory profiles and the outcomes of delivery of 54 APS patients from January 2000 to March 2013 were investigated retrospectively. RESULTS: (1) Maternal/fetal outcome: 17 pregnancies (31.4%) resulted in full term delivery, 7 (12.9%) in stillbirth, 16 (29.6%) in spontaneous abortion,10 (18.5%) in premature birth due to eclampsia or severe preeclampsia or signs of placental insufficiency, 4 (7.4%)received therapeutic termination of pregnancy due to eclampsia or severe preeclampsia. In 27 live birth cases, 8 (29.6%) were fetal growth restriction, 4 (14.8%) were low birth weight infants, and 3 (11.1%) were very low birth weight infants. (2) Influence of treatment on the pregnancy outcomes and complications: 24 APS patients were given the treatment of aspirin or aspirin combined with low molecular weight heparin, and 30 patients received no treatment. Compared with the untreated group, the treated group had lower rate of fetal loss, higher rate of full-term delivery, increased gestational age and birth weight, decreased incidence of preeclampsia / eclampsia and thrombocytopenia. There was a significant difference between the two groups (P<0.05). (3)Possible risk factors of unsuccessful pregnancy: there were 17 successful pregnancies and 37 unsuccessful pregnancy. The rate of double APL positive and antibody titers ≥ three times the upper limit of normal were higher in the unsuccessful pregnancy group than the successful pregnancy group. Antibody negative rate before pregnancy proportion of patients received treatment and the level of complement 4 were lower in the unsuccessful pregnancy group. CONCLUSION: Pregnant women with APS are an extremely high risk group for adverse maternal /fetal outcome. Treatments can improve the pregnancy outcome of the APS patients. APL not turning negative before pregnancy double APL positive, antibody titers ≥ three times the upper limit of normal and complement 4 decrease may be the risk factors for pregnancy failure and treatment may be a protective factor for successful pregnancy.

Efficient production of dihydroxyacetone from biodiesel-derived crude glycerol by newly isolated Gluconobacter frateurii.

The efficient production of dihydroxyacetone (DHA) on biodiesel-derived glycerol based media was developed. A newly isolated strain, Gluconobacter frateurii CGMCC 5397, could convert crude glycerol to DHA with high yield and productivity. In shake-flask fermentation, the DHA concentration of 73.1 gl(-1) was attained at 48 h using an optimum medium containing biodiesel-derived crude glycerol. When fed-batch fermentation was carried out in a 7-l stirred bioreactor with crude glycerol, the DHA concentration, productivity, and yield were 125.8 gl(-1), 2.6 gl(-1)h(-1), and 90.5% at 48 h, respectively. This study suggests that the inexpensive biodiesel-derived crude glycerol could be utilized for efficient production of DHA by G. frateurii.

[Enhancement pattern of thyroid carcinoma with contrast-enhanced ultrasound].

OBJECTIVE: To investigate the enhancement pattern of thyroid carcinoma with contrast-enhanced ultrasound (CEUS) for the diagnostic value. METHODS: Thirty-one cases with thirty-five thyroid Occupied were retrospectively reviewed. The final diagnosis was confirmed by biopsy pathology. Contrast agent SonoVue and contrast pulse sequencing (CPS) technique were used in this study. RESULTS: Thirty-five thyroid carcinoma presented three enhancement patterns with CEUS. Type I: twenty-three lesions enhanced in a pattern of ring with centripetal fill-in, however, the central part of no contrast agent filling. Type II: five lesions enhanced regularly and homogeneously. Type III: seven lesions enhanced irregularly and homogeneously. Thyroid benign and malignant lesions perfusion time curve, compared to papillary thyroid carcinoma with nodular goiter in the AT, TTP and WT indicators P < 0.05; papillary thyroid carcinoma and thyroid adenoma, compared to WT alone an indicator of P < 0.05. CONCLUSIONS: Identification of the different enhancement patterns of thyroid carcinoma could improve the diagnostic ability of CEUS.

Interventional effect of Ginkgo biloba extract on the progression of gastric precancerous lesions in rats.

OBJECTIVE: To investigate the effect of Ginkgo biloba extract on gastric precancerous lesions in rats. METHODS: 80 4-week-old Wistar rats were randomly divided into four groups: a control group, a model group, a low and a high dose Ginkgo biloba extract intervention group; 20 in each group. Gastric precancerous lesions were induced by giving them 100 mg/L N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) solution to drink ad libitum for 20 weeks. In addition to the MNNG, the intervention groups were lavaged with Ginkgo biloba extract (0.5 mg/kg/d in the low dose group, 1.5 mg/kg/d in the high dose group) for 20 weeks. Starting from week 21 all the rats were fed with normal rat chow and tap water. At the end of week 30 the rats were killed. The histopathological changes of their gastric mucosa, ISA, NGI, the serum and gastric mucosal SOD/MDA and the expressions of oncogenes were studied. RESULTS: The incidence of mild to severe intestinal metaplasia and dysplasia were significantly lower in the intervention groups than those in the model group (P < 0.01). The ISA and NGI in the intervention groups were significantly lower than those in the model group (P < 0.01). In the intervention groups the activity of SOD was increased and the concentration of MDA was decreased (P < 0.01). Expressions of Bcl-2, c-myc and FasL decreased in the intervention groups, whereas the expression of Fas increased. When compared with the model group, the differences were statistically significant (P < 0.01, P < 0.05, respectively). CONCLUSION: Ginkgo biloba extract can increase anti-oxidative activity and inhibit the progression of gastric precancerous lesions via the regulation of cell proliferation and apoptosis.

[Features of contrast-enhanced ultrasound imaging in malignant lymphoma after treatment with integrated Chinese and Western medicine].

OBJECTIVE: To investigate the typical features of contrast-enhanced ultrasound imaging in malignant lymphoma after treatment with integrated Chinese and Western medicine. METHODS: Ten patients with malignant lymphoma after different treatment cycles were examined with contrast-enhanced sonography, including 21 lymphnodes (in the neck, space of iliac blood vessels, retroperitoneum) and 7 spleen infiltrations. RESULTS: The blood flow pattern distribution in lymph node before freatment was significantly different from that after trentment showed either by color Doppler flow imaging (CDFI) or by contrast-enhanced ultrasound (CEUS, P < 0.05), and the difference in splenic lesion was shown by CEUS (P < 0.05). CONCLUSION: Contrast-enhanced ultrasound imaging is a valuable tool for the follow-ups of malignant lymphoma after chemotherapy and guiding subsequent treatment.

[Diagnosis of thyroid space-occupying lesions using real-time contrast-enhanced ultrasonography with sulphur hexafluoride microbubbles].

OBJECTIVE: To explore the value of contrast-enhanced ultrasound (CEUS) in the diagnosis of thyroid occupied lesions with injection of sulphur hexafluoride microbubbles. METHODS: Fifty nine cases of conventional ultrasonic diagnosis of thyroid lesions in patients with 73 lesions re-sulfur hexafluoride microbubble ultrasound contrast real-time inspection, the use of CEUS and contrast pulse sequencing (CPS). RESULTS: Seventy-three lesions were satisfied with the dynamic contrast perfusion imaging. Ultrasound contrast prompted the 15 lesions (13 cases) of malignant lesions by postoperative pathology confirmed papillary thyroid carcinoma, ultrasound images showed a low or mixed echo, boundary ambiguity, accompanied by Microcalcification foci, blood flow distribution I rank or grade III. Ultrasound prompted 58 lesions (46 cases) of benign lesions, blood flow distribution of grade II or III, the performance of a variety of two-dimensional ultrasound image, showing cystic, solid or liquid-solid mass, border clearance. Contrast with the surrounding thyroid tissue, lesionsor=20 mm papillary thyroid carcinoma, manifested prior to the beginning of the adjacent thyroid substance to enhance and dissection, showing the form of high-enhanced perfusion; thyroid follicular adenoma as early as the beginning of the adjacent thyroid substance to enhance and dissection, mass retained within the contrast agent longer time, showing the form of high-enhanced perfusion; nodular goiter enhanced for thyroid nodules and adjacent synchronization started to pick up in real terms, after the 6 lesions showed nodular contrast slightly earlier than the adjacent thyroid substance started to pick up, 28 lesions with synchronous real beginning of the adjacent thyroid dissection, 11 lesions of nodular dissipated early on (started receding time < or = 25 s) adjacent to thyroid substance. CONCLUSION: The real-time CEUS with injecting sulphur hexafluoride microbubbles is valuable in the diagnosis and differential diagnosis of the thyroid occupied lesions.

Generation of the transgenic potato expressing full-length spike protein of infectious bronchitis virus.

To seek a new delivery system of vaccine for infectious bronchitis virus (IBV), transgenic potato expressing full-length spike (S) protein of IBV was produced and its immunogenicity in chickens was investigated. One to three copies of S gene of IBV were randomly and stably inserted into potato (Solanum tuberrosum cv. Dongnong 303) genome by Agrobacterium tumefaciens-mediated transformation. Transcription and translation of S gene for IBV were confirmed by Northern blot and Western blot analyses in transgenic plantlets. Chickens immunized orally and intramuscularly with transgenic potato tubers expressing S protein generated the detectable levels of serum neutralizing antibodies and were protected against the challenge with the virulent IBV. In vitro secretion of interleukin 2 and T lymphocyte proliferation of spleen cells from the immunized chickens varied with the dose and the manner of vaccination with S protein derived from transgenic plants. The results indicated that S protein expressed in transgenic plants might be a new source for the production of Coronaviridae IBV vaccine.

Expression of immunogenic S1 glycoprotein of infectious bronchitis virus in transgenic potatoes.

The expression of infectious bronchitis virus (IBV) S1 glycoprotein in potatoes and its immunogenicity in mice and chickens were investigated. Potato plants were genetically transformed with a cDNA construct encoding the IBV S1 glycoprotein with the Agrobacterium system. Genomic DNA and mRNA analyses of the transformed plantlets confirmed the integration of the foreign cDNA into the potato genome, as well as its transcription. Mice and chickens vaccinated with the expressed IBV S1 glycoprotein produced antibodies that neutralized IBV infectivity. After three immunizations, vaccinated chickens were completely protected from virulent IBV infection. These results demonstrate that transgenic potatoes expressing IBV S1 glycoprotein can be used as a source of recombinant antigen for vaccine production.