Mycena, a symbiont of Gastrodia elata, promotes seed germination of G. elata and plays a crucial role in the sexual reproduction of G. elata. However, the lack of genetic transformation system of Mycena blocks the research on the interaction mechanism of the two. In order to establish the protoplast transformation system of Mycena, this study analyzed the protoplast enzymatic hydrolysis system, screened the resistance markers and regeneration medium, and explored the transient transformation. After hydrolysis of Mycena hyphae with complexes enzymes for 8 h and centrifugation at 4 000 r·min~(-1), high-concentration and quality protoplast was obtained. The optimum regeneration medium for Mycena was RMV, and the optimum resistance marker was 50 mg·mL~(-1) hygromycin. The pLH-HygB-HuSHXG-GFP-HdSHXG was transformed into the protoplast of Mycena which then expressed GFP. The established protoplast transformation system of Mycena laid a foundation for analyzing the functional genes of Mycena and the molecular mechanism of the symbiosis of Mycena and G. elata.
Agaricales ; Gastrodia/genetics* ; Protoplasts ; Symbiosis/genetics* ; Transformation, Genetic

Aim To explore the regulatory effect of Danggui-chuanxiong herb pair (GX) on JAK-STAT signaling pathway in rats with cerebral ischemia/reper-fusion injury (I/R). Methods The I/R injury rat model was constructed by modified suture occlusion method. After 24 hours of perfusion, Zea Longa scoring method was used to score the neurological function, TTC staining to detect the cerebral infarct volume of rats, HE staining to observe the pathological changes of brain tissues, the biochemical method to determine the MDA, SOD, GSH-Px expression, ELISA to detect the expression of NF-κB, VEGF, ICAM-1 and PAH in brain tissues, and immunohistochemical method to detect JAK2, p-STAT3, AKT And ERK1/2 expression of the brain tissue ischemic penumbra area. Results Compared with sham group, model rats had severe neurological damage, larger cerebral infarction, necrosis, edema, inflammation, disorder of nerve cell arrangement, abnormal cell enlargement, vacuole-like changes, neuron reduction and other pathologies in brain tissues. The expression JeveJs of MDA, NF-κB, VEGF, PAI-1 and ICAM-1 in brain tissues of model group significantly increased, and the expression levels of GSH-Px and SOD were significantly reduced. Compared with model group, the neurological scores of rats in GX

Objective:To investigate the prognostic significance of different IDH mutations and accompanying gene mutations in patients with non-M 3 acute myeloid leukemia (AML) . Methods:Second-generation sequencing was performed to detect the mutations of 22 genes in 389 patients with AML in the First Affiliated Hospital of Zhengzhou University from June 2016 to December 2018, and Kaplan-Meier and Cox regression models were used to analyze the prognostic factors.Results:The mutation frequency of IDH1 and IDH2 was 6.2% and 8.7% , respectively, in all patients without co-mutation. The IDH2 mutant group had an older age, higher proportion of bone marrow primitive cells, more common normal karyotype, and more common RUNX1 and SRSF2 mutations compared with IDH2 wild-type group. Univariate analysis of variance showed that the median OS and PFS of IDH1 mutation group were significantly shorter than those of the wild-type group ( P<0.05) . IDH2 mutation as a single variable and IDH2R140 mutation had no significant effect on the prognosis, while different mutation sites had different effects. Compared with the IDH2 wild-type group, the IDH2R172 mutation group had lower complete remission (CR) rate and shorter median OS and PFS ( P<0.05) . In patients with normal karyotypes or aged ≥50 years, IDH2 mutation as a single variable had no significant effect on the prognosis, IDH1 mutation and IDH2R172 mutation were associated with poor OS and PFS ( P<0.05) , and IDH2R140 mutation had no significant effect on OS and PFS. Approximately 74.1% (43/58) of patients with IDH mutation simultaneously carried other gene mutations; however, the number of accompanying gene mutations had no significant effect on the prognosis. Among 58 patients with IDH mutation, the CR rate of patients with NPM1 mutation was significantly higher than that of patients in the NPM1 wild-type group (81.8% vs 36.4% , P=0.014) , the median OS in patients with DNMT3A mutation was lower than that of patients with DNMT3A wild type [4.0 months (95% CI 3.8-4.2) vs 6.3 months (95% CI 2.4-10.2) , P=0.041) ]. Multivariate analysis showed that age ≥60 years and white blood cell count ≥100×10 9/L were independent risk factors for OS and PFS, while CR after two courses of treatment and hematopoietic stem cell transplantation were independent prognostic favorable factors for OS and PFS. Conclusion:In patients with AML (non-M 3) , IDH gene mutations often coexisted with other gene mutations, and different subtypes and accompanying gene mutations of IDH have different prognostic significance.

INTRODUCTION: Family history of psychopathology is a risk factor for mood and anxiety disorders in children, but little is known about rates of parental psychopathology among treatment-seeking youth with affective disorders in the Asia Pacific region. This study examined patterns of emotional and behavioural problems in parents of clinically-referred youth in Singapore. We hypothesised that parents would have higher rates of affective disorders compared to the Singapore national prevalence rate of 12%. MATERIALS AND METHODS: In this cross-sectional study, 47 families were recruited from affective disorders and community-based psychiatry programmes run by a tertiary child psychiatry clinic. All children had a confirmed primary clinical diagnosis of depression or an anxiety disorder. Parents completed the Mini International Neuropsychiatric Interview (MINI) to assess for lifetime mood and anxiety disorders. They also completed the Adult Self Report (ASR) and Adult Behavior Checklist (ABCL) to assess current internalising and externalising symptoms. RESULTS: Consistent with our hypothesis, 38.5% of mothers and 10.5% of fathers reported a lifetime mood and anxiety disorder. Nearly 1/3 of mothers had clinical/subclinical scores on current internalising and externalising problems. A similar pattern was found for internalising problems among fathers, with a slightly lower rate of clinical/subclinical externalising problems. CONCLUSION Our findings are consistent with previous overseas studies showing elevated rates of affective disorders among parents - particularly mothers - of children seeking outpatient psychiatric care. Routine screening in this population may help to close the current treatment gap for adults with mood and anxiety disorders.
Adult ; Anxiety Disorders ; diagnosis ; epidemiology ; psychology ; Child ; Cross-Sectional Studies ; Family Health ; statistics & numerical data ; Female ; Humans ; Male ; Mood Disorders ; diagnosis ; epidemiology ; psychology ; Parent-Child Relations ; Parenting ; psychology ; Parents ; psychology ; Psychiatric Status Rating Scales ; Psychopathology ; Singapore ; epidemiology

Objective: To investigate the anti-inflammatory effect of hesperetin (HSP) on lung damage induced by paraquat (PQ) in rats by detecting the levels of inflammatory makers in rat lung tissues. Methods: 140 Wistar male rats were randomly divided into negative control group, HSP control group, HSP control group, paraquat model group, pirfenidone (PDF) positive control group, and 100, 200, 400 mg/kg HSP treatment groups. All groups were exposed to 50mg/kg paraquat by oral gavage except for the negative control group and HSP control group. After 24 hours, the rats in each group were given drug intervention once daily. 10 rats were randomly sacrificed at 7th day and 28th day after exposure to paraquat respectively. 3 rats were randomly selected from them and HE, Masson staining were used to observe the pathological changes in the lungs of each group. Each group randomly selected 6 rats at two time points to detect the levels of TGF-β₁, TNF-α, IL-4, IL-10, IL-1β and IFN-γ in rat lung tissues. Results: Histopathological examination found that the lung injury were reduced in the rats of PDF positive control group and all HSP treatment groups. Compared with the negative control group, the levels of TGF-β1, IL-1β, TNF-α, IL-4, and IL-10 in rat lung tissues were significantly increased (P<0.05, P<0.01) after PQ exposure at two points in time, and there was no significant difference in the level of IFN-γ in lung tissues compared with the negative control group (P>0.05) . The levels of TGF-β1, IL-1β, IL-4, IL-10 and TNF-α in the lung tissues of rats on the 7th day in different dose treatment groups of HSP were reduced compared with those in the PQ model group with varying degrees (P<0.05, P<0.01) . The level of IFN-γ in lung tissues of rats were not significantly different from that of model group (P>0.05) . The levels of TGF-β₁ and TNF-α in lung tissue of rats on the 28th day in PDF positive control group and different dose treatment groups of HSP were reduced compared with those in the PQ model group with varying degrees (P<0.05, P<0.01). The levels of IFN-γ in the rat lung tissues were increased compared with those in the PQ model group (P<0.05). Besides, there were no significant in the levels of IL-1β, IL-4 and IL-10 in lung tissues compared with PQ model group (P>0.05). Conclusion HSP can reduce lung damage induced by PQ in rats by inhibiting the release of inflammatory factors and promoting the secretion of anti-inflammatory factors.

Estrogens are commonly used in gynecologic area, such as oral contraception, hormone replacement therapy, and in vitro fertilization-embryo transfer. Although estrogen is a common cause of acute drug-induced pancreatitis, there has been paucity of report in Korea. Clinical course of estrogen-induced acute pancreatitis is usually mild to moderate, but fetal case can occur. In addition, there can be a latency from the first administration to the symptom. Therefore, physicians should consider the possibility of the disease when a woman taking estrogen or previous history of taking estrogen presents with acute abdominal pain. Here, we report a case of estrogen-induced acute pancreatitis that occurred during the preparation for embryo transfer.
Abdominal Pain ; Contraception ; Embryo Transfer ; Estrogens ; Female ; Hormone Replacement Therapy ; Humans ; In Vitro Techniques ; Korea ; Pancreatitis*

objective To explore the role of Xanthatin in the targets of epithelial-mesenchymal transition (EMT) process using molecular docking method,and the effect on the target protein expression of HepG2 cells was detected by Western assay.Method Dhs,Vimentin,Snail and VEGFR3 are critical targets in EMT process,the spatial binding ability of Xanthium was evaluated by molecular docking method,compared with the corresponding endogenous substances:nicotinarnide adenine dinucleotide,Acetate ion,flavin adenine dinucleotide,and N-Acetyl-D-glucosamine.HepG2 cells were cultured,and the effects of Xanthatin of 1,5 and 20 mol/L concentrations on Dhs,Vimentin,Snail and VEGFR3 protein expression were detected by Western Blotting assay.Rusult Molecular docking show that Xanthatin has obvious affinity to key factors of EMT process such as Dhs,Vimentin,and VEGF-R3,higher than that of endogenous substance;and the affinity with Vimentin was less than that of endogenous substance;Western Blotting experiments proved the virtual results.The expression of Vimentin,Snail,VEGFR3 protein was significantly lowered,and the expression of e-cadherin was significantly raised.Conclusion The influence of Xanthatin to key factor e-cadherin,Vimentin,Snail,VEGFR3 are obvious,Which is likely to be a potential target.The results of computer virtual experiment and Western Blotting have certain similarity.Molecular virtual docking can pre hint the potential target factor.

Clonorchis sinensis is a Group-I bio-carcinogen, associated with cholangiocarcinoma (CCA). The hamster is the only experimental model of C. sinensis-mediated CCA, but we oblige another animal model. The present study intended to develop a C. sinensis (Cs) mediated CCA model using C3H/He mice, co-stimulated with N-nitrosodimethyl-amine (NDMA) and dicyclanil (DC). The mice were divided into 8 groups with different combinations of Cs, NDMA, and DC. Six months later the mice were sacrificed and subjected to gross and histopathological examination. The body weights were significantly reduced among the groups treated with 2 or more agents (eg. Cs+NDMA, Cs+DC, NDMA+DC, and Cs+NDMA+DC). In contrast, liver weight percentages to body weight were increased in above groups by 4.1% to 4.7%. A Change of the spleen weight was observed only in Cs+NDMA group. Though C. sinensis infection is evident from hyperplastic changes, only 1 worm was recovered. T wo mice, 1 from Cs and the other from Cs+DC group, showed mass forming lesions; 1 (281.2 mm3) from the Cs group was a hepatocellular adenoma and the other (280.6 mm3) from the Cs+DC group was a cystic mass (peliosis). Higher prevalence of gray-white nodules was observed in Cs group (42.9%) followed by Cs+NDMA+DC group (21.4%). The mice of the Cs+NDMA+DC group showed hyper-proliferation of the bile duct with fibrotic changes. No characteristic change for CCA was recognized in any of the groups. In conclusion, C3H/He mice produce no CCA but extensive fibrosis when they are challenged by Cs, NDMA, and DC together.
Adenoma, Liver Cell ; Animals ; Bile Ducts ; Body Weight ; Cholangiocarcinoma* ; Clonorchis sinensis* ; Cricetinae ; Dimethylnitrosamine* ; Fibrosis ; Liver ; Mice* ; Models, Animal ; Models, Theoretical ; Prevalence ; Spleen

Objective To explore the differentiated diagnostic value of the morphological changes of follicular dendritic cell (FDC)meshwork between different subtype of lymphoma.Methods CD21 was stained by immunohistochemistry (IHC) method,FDC meshwork pattern was studied in 5 6 cases of various lymphomas(including 8 cases of diffuse large B cell lym-phoma,2 cases of burkitts lymphoma,6 cases of small lymphocytic lymphoma,6 cases of plasmacytoma,3 cases of MALT lymphoma,6 cases of peripheral T cell lymphoma,3 cases of anaplastic large cell lymphoma,8 cases of NK/T cell lympho-ma,4 cases of follicular lymphoma,3 cases of mantle cell lymphoma,3 cases of AITL,2 cases of FDC sarcoma).Results FDC meshwork in the morphological changes of various subtypes of lymphoma could be classified into four patterns:①FDC form a disappeared and disintegrated meshwork,most of the lymphoma FDC meshwork fully or partially destroyed,including diffuse large B cell lymphoma,burkitt lymphoma,small lymphocytic lymphoma,plasmacytoma,peripheral T cell lymphoma, anaplastic large cell lymphoma,NK/T cell lymphoma;②FDC meshtwork existence,even hyperplasia,including follicular lymphoma,mantle cell lymphoma,MALT lymphoma;③FDC meshtwork proliferation,disorder and deformation,such as AI-TL;④Full expression subtype:such as FDC sarcoma.Conclusion The morphologic pattern of the FDC meshwork in lym-phomas of follicular origin was differs according to the lymphoma subtypes,and it has important clinical value in lymphoma differential diagnostic.

Objective To investigate the quality of life and its influencing factors of families primary caregivers of dementia patients .Methods The typically self made questionnaire ,WHO quality of life measurement profile (WHOQOL‐BREF) ,zarit bur‐den of care scale(ZBI) ,social support rating scale (SSRS) et al were adopted to investigate 64 caregivers of AD patients .Results (1) The overall quality of life of family caregivers of total subjective feelings were significantly lower than the general health status of subjective feeling score (t= -2 .217 ,P=0 .030) .The field of physiological and psychology points have no statistically significant difference in comparison with Chinese norm (t= -0 .252 ,-1 .994 ,P=0 .801 ,0 .050);the field of social and environment points have statistically significant difference compared with norm(t= -3 .830 ,3 .658 ,P=0 .000 ,0 .001) .(2) caregiver who have spouse , poor parent child relationship and patients with somatic diseases and mental behavior symptoms have lower quality of life(P<0 .05) .(3)Total score of Quality of life was negatively associated with total score of care burden (r= -0 .433 ,P=0 .000) ,while was significant positive associated with total score of the social support (r=0 .346 ,P=0 .005) .(4)Marital status ,parent‐child rela‐tionship ,physical illness and social support were the main factors affecting the quality of life(r2 =0 .409) .Conclusion Strengthen the social support and psychological intervention of dementia caregivers who has a spouse ,poor parent-child relationship and poor physical condition may help to improve the quality of life .