Objective To explore the regulatory role of farnesol in Candida albicans(C.albicans)biofilm cAMP-PKA signaling pathway and its correlation with drug resistance.Methods Standard,fluconazole-resistant,wild and high RAS1 gene expression strains of C.albicans were cultured to different phases of the biofilm(6,12,24,36 h),and the sessile minimal inhibitory concentration 50％(SMIC50)of fluconazole were determined by XTT reduction after farnesol treatment.The regulatory effects of farnesol on the ex-pression of genes related to the cAMP-PKA signaling pathway in standard and fluconazole-resistant strains of C.albicans,such as RAS1,CYR1,PDE2 were examined using qPCR;the effects of farnesol on the protein expression of the pathway were analyzed by Western blot.RAS1 gene expression of the wild and high RAS1 gene expression strains was measured by qPCR.Results ① Compared with the standard strain,resistant strains of C.albicans had higher levels of biofilm SMIC50 at 6,12 and 24 h;there was no significant difference in RAS1 expression(P＞0.05),while CYR1 expression increased significantly at 6 and 24 h in the biofilm(P＜0.01),and PDE2 expression decreased at 6 h in the biofilm(P＜0.01).②After treatment with farnesol,the resistance of the biofilm of the standard strain and drug-resistant strain decreased.Compared with no treatment with farnesol,the expression of RAS1 in the biofilm of the standard strain and drug-resistant strain decreased at all time points(P＜0.01);CYR1 expression decreased in the biofilm at 6,24 and 36 h,and in-creased in the biofilm at 12 h(P＜0.01);PDE2 expression increased in the 12 h biofilm(P＜0.01).③Compared with the wild strain,the high expression strain of RAS1 gene showed higher SMIC50 in the biofilm at 12 and 24 h,and significantly higher expression of RAS1 gene in the biofilm at 12,24 and 36 h(P＜0.01).④After treatment with farnesol,the resistance of wild-type strains and high expres-sion strains of RAS1 gene decreased.Compared with the untreated group,the expression of RAS1 gene in the biofilm of wild-type and RAS1 gene high expression strain decreased at 12 and 24 h(P＜0.01).Conclusion Farnesol can affect the sensitivity of C.albicans biofilm to fluconazole by regulating the expression of resistance molecules RAS1,CYR1 and PDE2 in the cAMP-PKA pathway.The regulatory effect varies at different stages of biofilm formation.

Objective To explore the regulatory role of farnesol in Candida albicans(C.albicans)biofilm cAMP-PKA signaling pathway and its correlation with drug resistance.Methods Standard,fluconazole-resistant,wild and high RAS1 gene expression strains of C.albicans were cultured to different phases of the biofilm(6,12,24,36 h),and the sessile minimal inhibitory concentration 50％(SMIC50)of fluconazole were determined by XTT reduction after farnesol treatment.The regulatory effects of farnesol on the ex-pression of genes related to the cAMP-PKA signaling pathway in standard and fluconazole-resistant strains of C.albicans,such as RAS1,CYR1,PDE2 were examined using qPCR;the effects of farnesol on the protein expression of the pathway were analyzed by Western blot.RAS1 gene expression of the wild and high RAS1 gene expression strains was measured by qPCR.Results ① Compared with the standard strain,resistant strains of C.albicans had higher levels of biofilm SMIC50 at 6,12 and 24 h;there was no significant difference in RAS1 expression(P＞0.05),while CYR1 expression increased significantly at 6 and 24 h in the biofilm(P＜0.01),and PDE2 expression decreased at 6 h in the biofilm(P＜0.01).②After treatment with farnesol,the resistance of the biofilm of the standard strain and drug-resistant strain decreased.Compared with no treatment with farnesol,the expression of RAS1 in the biofilm of the standard strain and drug-resistant strain decreased at all time points(P＜0.01);CYR1 expression decreased in the biofilm at 6,24 and 36 h,and in-creased in the biofilm at 12 h(P＜0.01);PDE2 expression increased in the 12 h biofilm(P＜0.01).③Compared with the wild strain,the high expression strain of RAS1 gene showed higher SMIC50 in the biofilm at 12 and 24 h,and significantly higher expression of RAS1 gene in the biofilm at 12,24 and 36 h(P＜0.01).④After treatment with farnesol,the resistance of wild-type strains and high expres-sion strains of RAS1 gene decreased.Compared with the untreated group,the expression of RAS1 gene in the biofilm of wild-type and RAS1 gene high expression strain decreased at 12 and 24 h(P＜0.01).Conclusion Farnesol can affect the sensitivity of C.albicans biofilm to fluconazole by regulating the expression of resistance molecules RAS1,CYR1 and PDE2 in the cAMP-PKA pathway.The regulatory effect varies at different stages of biofilm formation.

Depression is a major cause of disability and has adverse effects.Despite the many types of anti-depressants,clinical treatments of depression remain poor.Therefore,novel anti-depressant mechanisms need to be explored.The beneficial effects of irisin on the nervous system are gradually being elucidated,and studies have found that irisin has an anti-depressant effect,which may become a new treatment for depression.This study explored the mechanism of irisin and its upstream and downstream anti-depressants by reviewing the existing studies explaining the link between irisin and depression,and proposes that SIRT1/PGC-1α may mediate FNDC5/irisin to regulate BDNF to promote neurogenesis and improve depression,which provides a new idea to study irisin and its upstream and downstream anti-depressants.

Irritable bowel syndrome （IBS） is a functional gastrointestinal disease， but it often causes extreme gastrointestinal discomfort and prolonged illness， which seriously affects the quality of life of patients. The global incidence rate is increasing year by year. Clinically， western medicine mainly uses oral antispasmodics， secretagogues， and antidepressants， but there are many disadvantages such as adverse reactions and poor long-term efficacy. Therefore， finding an efficient and safe treatment method is an urgent problem to be solved. A large number of studies have shown that traditional Chinese medicine has definite curative and long-lasting effects on the treatment of IBS， which has become a hot research direction in recent years. By searching Chinese and foreign literature， it is found that electroacupuncture， moxibustion， Chinese medicine monomers， and compound decoctions are the main methods in the mechanism research of traditional Chinese medicine in the treatment of IBS-related pathways， and their signaling pathways involve nuclear transcription factor kappa B （NF-κB）， transient receptor potential vanillin subfamily 1 （TRPV1）， 5-hydroxytryptamine （5-HT）， mitogen-activated protein kinase （MAPK）， and so on. Traditional Chinese medicine can repair intestinal inflammation， reduce visceral sensitivity， enhance intestinal mucosal barrier， and regulate intestinal motility by regulating this series of signaling pathways， thereby playing an important role in the treatment of IBS with multi-level， multi-link and multi-target characteristics. Based on the cell signaling pathways， this paper reviewed the research progress on the mechanism of traditional Chinese medicine in the treatment of IBS， hoping to provide theoretical support and diagnosis and treatment ideas for the clinical treatment of IBS with traditional Chinese medicine.

Objective: To study the clinicopathological features, diagnostic features and differential diagnoses of SMARCB1 (INI1)-deficient sinonasal carcinoma (SDSC). Methods: Six cases of SDSC diagnosed at Eye, Ear, Nose and Throat Hospital, Fudan University from 2016 to 2018 were retrieved; the clinical features, histomorphology, immunophenotype, radiology and outcome were analyzed with review of literature. Results: There were five men and one woman with age range of 37 years to 75 years (mean 56 years). One case was in stage T2, and 5 cases were in stage T4. Computer tomography and magnetic resonance imaging showed a mass occupying the sinonasal cavity with bone destruction in all six patients. Microscopically, the tumors had infiltrative margins. Four tumors were composed mostly of basaloid cells, which possessed high nuclear/cytoplasmic ratio,scant cytoplasm,and minimalnuclear pleomorphism; and the cells were arranged in sheets or nests in a desmoplastic stroma. Two tumors were composed of rhabdoid cells, which possessed abundant, eosinophilic cytoplasm and eccentric nuclei, often growing in a nests or sheets pattern. Immunohistochemical staining showed that 6/6 cases had complete loss of INI1, diffusely and strongly positive for CKpan, and were negative for S-100 and EBER ISH; 4/6 cases were focally positive for p63; 1/5 was focally positive for Syn and p16. The Ki-67 index was 30% to 70%. The follow-up period ranged 1-26 months, with one patient died of extensive metastases, one had local recurrence, and two had lymph node metastases; one was alive without disease, and one was lost to follow-up. Conclusions SMARCB1 (INI1)-deficient sinonasal carcinoma is mostly aggressive, with rapid progression and poor prognosis. Histomorphological spectrum predominantly consists of basaloid type and rhabdoid type. The complete loss of nuclear expression of INI1 can help to distinguish this tumor from its many mimickers.

Objective To analyze the risk factors of post-stroke disability in patients with acute cerebral infarction.Methods Total 1 137 consecutive patients with acute cerebral infarction admitted in Department of Neurology, General Hospital of Jinan Military Region, were prospectively recruited from August 2010 to August 2014.According to Oxfordshire Community Stroke Project(OCSP), 275 patients were classified as total anterior circulation infarction, 377 as partial anterior circulation infarction,305 as posterior circulation infarction and 180 as lacunar infarction.The baseline data including age, gender, National Institute of Health Stroke Scale ( NIHSS) score were recorded.Recovery was assessed by modified Rankin Scale ( mRS) 6 months after stroke by telephone interview ( mRS≤2:good prognosis, 2

Objective To introduce the operation skills of tail vein injection in conscious rats , and improve the success rate of ingection .Methods The rat was fixed by the operator with a self-made binding clothes , one person assists to fix the rat tail, one person performs puncture , and one person performs injection .During the injection process , we should minimize the injury to the rat tail and the stress caused by operation , strictly limit the needle point , the number of puncture , the maximum dose and injection speed , and to make the needle position away from the injection site to avoid contamination and waste of the drug solution .Results The operation method was successfully established and it was fast , stable, with good repeatability and high degree of coordination .Conclusion This operation is rapid, reliable and stable, worthy of recommendation , especially for the intravenous injection of expensive drugs .

Objective To investigate the effect of saw palmetto extract (SR) on GL261 glioma in rats and immune system.Methods The 40 rats were divided into 4 groups randomly,one was the control group without tumor (n =10),the other 30 rats were given subcutaneous inoculation of tumor cells and then divided into 3 groups:tumor-bearing group (n =10),low dose SR group (n =10),and high dose SR group (n =10).After 1 weeks feeded,the rats of SR groups were given the saw palmetto extract,low dose group 50 mg/kg once a day every other day and 300 mg/kg of high dose group every other day.The tumor-bearing groups received the same dose of distilled water.After 4 weeks feeding,we measured the tumor weight and the inhibition rate was calculated.The tumor cell apoptosis was detected by TdT mediated dUTP nick-end labeling (TUNEL) staining.The splenic T lymphocyte proliferation was tested by methyl thiazolyl tetrazolium (MTF).Results (1) The SR groups compared to tumor group,the tumor weight was significantly reduced (F =62.678,P =0.000).The tumor inhibition rate was significantly higher in high dose group.(2) The apoptosis of tumor cells in tumor-bearing group was significantly less than SR groups and the apoptosis was significantly increased after treatment with SR,especially in high dose SR group (F =1.287E3,P =0.000).(3) Compared to SR groups and control group,T lymphocyte proliferation of tumor-bearing group reduced obviously.After treated with SR,T lymphocyte proliferation significantly increased and was higher in high-dose group (F =103.565,P =0.000).Conclusions Saw palmetto extract can activate T lymphocytes and induce apoptosis to tumor cells.Its function was related to SR concentration.

Objective To investigate the correlation between anemia and outcome in a large cohort of unselected patients with acute cerebral infarction.Methods Consecutive acute cerebral infarction patients who were hospitalized were prospectively recruited from August 2010 to November 2013.Eight hundred and fifty-eight patients were enrolled,and the baseline data including age,sex,National Institute of Health Stroke Scale(NIHSS) scores,type of Oxfordshire Community Stroke Project(OCSP:total anterior circulation infarct,partial anterior circulation infarct,posterior circulation infarct and lacunar infarct),serum creatinine,initial hemoglobin level,initial hematocrit level,etc,were recorded.Hemoglobin level and hematocrit level during hospitalization were also recorded.Domestic criteria were used to define if the patient had anemia on admission.Recovery was assessed by modified Rankin Scale (mRS) 180 days after stroke by telephone interview (mRS scores ≤ 2 reflected good prognosis,and mRS scores ＞ 2 reflected unfavorable prognosis).The influence on outcome by anemia on admission,initial hemoglobin level,nadir hemoglobin level,nadir hematocrit level was analyzed by multinomial Logistic regression analysis.Results Odds ratio of initial hemoglobin level for poor outcome was 1.013 (95％ CI 1.001-1.024,P =0.027) with each decrease in hemoglobin of 1 g/dl.Initial anemia(OR =2.417,95％ CI 1.202-4.859,P =0.013) was a independent prognostic factor for mortality;odds ratio of nadir hemoglobin level for mortality was 1.016(95％ CI 1.002-1.030,P =0.026) with each decrease in hemoglobin of 1 g/dl;odds ratio of nadir hematocrit level for mortality was 1.047(95％ CI 1.003-1.093,P =0.037) with decrease in hematocrit of one percentage point.Conclusions Initial hemoglobin level was a independent prognostic factor for poor outcome in patients with acute cerebral infarction.Anemia on admission,nadir hemoglobin level,nadir hematocrit level were independent prognostic factors for mortality in patients with acute cerebral infarction.

Objective To investigate the correlation between prestroke comorbidity and long-term outcomes in patients with acute cerebral infarction.Methods Consecutive acute ischemic stroke patients who were hospitalized were prospectively recruited from August 2010 to November 2012.Six hundred and forty-four patients were enrolled,the baseline data including Charlson Comorbidity Index (CCI),National Institute of Health Stroke Scale (NIHSS) score,type of Oxfordshire Community Stroke Project (OCSP:total anterior circulation infarct,partial anterior circulation infarct,posterior circulation infarct and lacunar infarct) were recorded.And recovery was assessed by modified Rankin Scale (mRS) 90 days after stroke by telephone interview (mRS score ≤ 2 reflected good prognosis,and mRS score ＞ 2 reflected unfavorable prognosis).Because CCI included specific comorbidity,we considered CCI,CCI without specific comorbidity and specific comorbidity as variable respectively.After screening the risk factors affecting prognosis using univariate analysis,the relationship between comorbidity and prognosis was estimated using multinomial logistic regression model.Results CCI was an independent predictor of good prognosis and unfavorable prognosis (OR =3.446,95％ CI 1.662-7.417; P =0.001).Congestive heart failure and diabetes were each independent predictor of good prognosis and unfavorable prognosis also (diabetes:OR =2.584,95％ CI 1.709-3.906,P =0.000; congestive heart failure:OR =6.229,95％ CI 1.705-22.755,P =0.006).Conclusions After acute ischemic stroke,the patients with the higher CCI score,diabetes and congestive heart failure are more likely to achieve unfavorable outcome.CCI,diabetes and congestive heart failure can each be used as a sensitive index to evaluate the 90 d prognosis of patients.Trial registration Clinical Research Center of China (CHiCTR-OCH-14004228)