Fatal and Rapid Progressive Isolated Cerebral Mucormycosis Involving the Bilateral Basal Ganglia: A Case Report.

Isolated cerebral mucormycosis is a clinical type of mucormycosis that is estimated to account for 8% of all mucormycosis cases. The clinical symptoms of isolated cerebral mucormycosis are elusive, and thus conventional techniques often lake sensitivity and specificity. Moreover, cultures are often negative, even when direct microscopy examination is positive. Although histopathology will probably remain the gold standard for the diagnosis of mucormycosis, obtaining a biopsy specimen is not always feasible in most vulnerable populations. Thus, molecular approaches are currently used as an advantageous assistant examination method to improve the early identification of the causative agent and subsequently guide therapy to improve the prognosis of patients. Here, we report a case of isolated cerebral mucormycosis caused by Rhizopus microspores in a healthy young adult that was identified using next-generation sequencing technology.

The excretory-secretory products of Echinococcus granulosus protoscoleces directly regulate the differentiation of B10, B17 and Th17 cells.

BACKGROUND: Excretory-secretory products (ESPs) released by helminths are well-known to regulate T cell responses in the host. However, their direct influence in the differentiation of naïve T cells, and especially B cells, remains largely unknown. This study investigated the effects of Echinococcus granulosus protoscoleces ESPs (EgPSC-ESPs) on the differentiation of IL-10-producing B cells (B10), IL-17A-producing B cells (B17) and Th17 cells. METHODS: BALB/c mice injected with EgPSC were used to evaluate the in vivo profiles of B10, B17 and Th17 cells. In vitro purified CD19+ B and naïve CD4+ T cells were cultured in the presence of native, heat-inactivated or periodate-treated EgPSC-ESPs, and the differentiation of these cell subsets were compared. RESULTS: In contrast to the control group, infected mice showed higher frequencies of B10, B17 and Th17 cells, and higher levels of IL-10 and IL-17A in the sera. Interestingly, B17 cells were first identified to express CD19+CD1dhigh. In vitro, B cells cultured with native ESPs exhibited a higher percentage of B10 cells but lower percentage of B17 and Th17 cells compared to the PBS group. Moreover, the relative expression of IL-10 and IL-17A mRNA were consistent with the altered frequencies. However, ESPs subjected to heat-inactivation or periodate treatment exhibited an inverse effect on the induction of these cell subsets. CONCLUSIONS: Our findings indicate that ESPs released by EgPSC can directly regulate the differentiation of B10, B17 and Th17 cells, which appear to be heat-labile and carbohydrate-dependent.

Escitalopram attenuates ß-amyloid-induced tau hyperphosphorylation in primary hippocampal neurons through the 5-HT1A receptor mediated Akt/GSK-3ß pathway.

Tau hyperphosphorylation is an important pathological feature of Alzheimer's disease (AD). To investigate whether escitalopram could inhibit amyloid-ß (Aß)-induced tau hyperphosphorylation and the underlying mechanisms, we treated the rat primary hippocampal neurons with Aß1-42 and examined the effect of escitalopram on tau hyperphosphorylation. Results showed that escitalopram decreased Aß1-42-induced tau hyperphosphorylation. In addition, escitalopram activated the Akt/GSK-3ß pathway, and the PI3K inhibitor LY294002 blocked the attenuation of tau hyperphosphorylation induced by escitalopram. Moreover, the 5-HT1A receptor agonist 8-OH-DPAT also activated the Akt/GSK-3ß pathway and decreased Aß1-42-induced tau hyperphosphorylation. Furthermore, the 5-HT1A receptor antagonist WAY-100635 blocked the activation of Akt/GSK-3ß pathway and the attenuation of tau hyperphosphorylation induced by escitalopram. Finally, escitalopram improved Aß1-42 induced impairment of neurite outgrowth and spine density, and reversed Aß1-42 induced reduction of synaptic proteins. Our results demonstrated that escitalopram attenuated Aß1-42-induced tau hyperphosphorylation in primary hippocampal neurons through the 5-HT1A receptor mediated Akt/GSK-3ß pathway.

[T Follicular Helper Cells and Related Molecules in Schistosoma japonicum Infected Mice after Praziquantel Treatment].

Objective: To investigate the pathological changes of liver and spleen of mice infected with Schistosoma japonicum and the changes of T follicular helper (Tfh) cells and surface molecules after praziquantel treatment. Methods: Fifteen female C57BL/6 mice （6-8 weeks） were randomly assigned into the praziquantel treated infection group （treated group）, infection control group (untreated group) and uninfected group （n=5 in each group）. The mice in the treated group and untreated group were each infected with 20 S. japonicum cercariae through the abdominal skin, and mice in the treated group were further administered with intragastric praziquantel [200 mg/（kg·d）] at week 6 post-infection for 3 consecutive days. Mice were sacrificed at week 4 after treatment to observe the morphological changes of liver and spleen and calculate the worm reduction rate and the liver egg reduction rate. The Tfh cell to CD4+ T cell ratio, as well as the expression of inducible T-cell costimulator （ICOS） and programmed cell death protein 1（PD-1） on peripheral blood and spleen, were determined by flow cytometry. Schistosome soluble egg antigen (SEA) specific IgG antibodies in serum were detected by ELISA. Results: The pathological changes of liver and spleen in the treated group were less severe compared with those of the untreated group, with a worm reduction rate of 84.1% and liver egg reduction rate of 69.1%. Flow cytometry showed that the percent of Tfh cells in peripheral blood and spleen was significantly higher in the treated group（14.7%-18.0%, 15.6%-25.0%） and the untreated group（13.7%-16.7%, 12.4%-18.2%） than that in the uninfected group（2.5%-6.8%, 4.9%-8.0%）, but there was no significant difference between the treated and untreated groups. The expression of ICOS in the peripheral blood and the spleen was significantly higher in untreated group（1.3%-3.2%, 4.1%-7.0%） than in the treated group（0.7%-1.1%, 1.8%-6.8%） and the uninfected group（0.2%-0.3%, 0.5%-0.8%）（P<0.01）, The expression of ICOS in the spleen was significantly higher in the treated group than in the uninfected group (P<0.01), while this difference was not found for ICOS expression in the peripheral blood. The PD-1 expression in the peripheral blood and spleen was significantly higher in the untreated group（0.8%-1.9%, 4.1%-10.7%） than in the uninfected group（0.4%-0.8%, 1.2%-1.8%）（P<0.01）, while there was no significant difference between the treated group（0.5%-1.5%, 4.5%-8.9%） and the untreated group (P>0.05). In addition, there was no significant difference in the level of SEA specific IgG between the treated group（2.015±0.061） and the untreated group（1.969±0.038） at 4 weeks after praziquantel treatment. Conclusion: Praziquantel treatment can significantly alleviate the lesions of the liver and the spleen and decrease the ICOS expression by Tfh cells in the peripheral blood and spleen.

[Serological Analysis of Antibodies against Cysticercus cellulosae and Toxoplasma gondii in 122 Patients with Meningitis Encephalitis Syndrome].

Objective: To examine the IgG and IgM antibodies for parasites Cysticercus cellulosae and Toxoplasma gondii in 122 patients with meningitis encephalitis syndrome, and provide basis for clinical diagnosis of the meningitis encephalitis syndrome. Methods: The sera were collected from patients with meningitis encephalitis syndrome in Shanghai Jiaotong University Affiliated Sixth People's Hospital, People's Hospital of Danyang City, and Jiangsu University Affiliated Hospital from August, 2014 to December, 2015. Serum IgG and IgM antibodies for cysticercus and T. gondii were examined using antibody test kits. The antibody positive rate was calculated and its distribution was analyzed by gender, season, age and occupation. Results: A total of 122 patients with meningitis encephalitis syndrome were included. Seventeen and 22 patients of them were positive for IgG （13.9%, 17/122） and IgM（18.0%, 22/122） against cysticercus, respectively, while 29 and 8 cases were positive for IgG （23.8%, 29/122） and IgM （6.6%, 8/122） against T. gondii. The positive rate of cysticercus and T. gondii in males was 30.6%（22/72） and 31.9%（23/72） respectively, while that in females was 26.0%（13/50） and 24.0% （12/50）. The positive rate of IgM against cysticercus was 12.0%（3/25）, 27.0%（17/63）, 6.9% （2/29）, and 0（0/5） from spring to winter, highest within 13-25 years（45.0%, 9/20） among age groups, and highest in workers（7/14） among various occupations. The positive rate of IgM against T. gondii was 4.0%（1/25）, 11.1% （7/63）, 0（0/29）, and 0（0/5） from spring to winter, highest in ages >65 years（44.0%, 11/25）, and highest in patients with other occupations（4/10）. There was no statistically significant difference in the positive rate between males and females, and among different seasons, ages and occupations. Conclusion: The positive rate of antibodies against cysticercus and T. gondii is high in the patients included, suggesting that a serological test for parasite infection might be performed during clinical diagnosis of meningitis encephalitis syndrome.

Escitalopram Ameliorates Tau Hyperphosphorylation and Spatial Memory Deficits Induced by Protein Kinase A Activation in Sprague Dawley Rats.

Here, we investigated the effect of escitalopram pretreatment on protein kinase A (PKA)-induced tau hyperphosphorylation and spatial memory deficits in rats using western blot and behavioral tests, respectively. We demonstrated that escitalopram effectively ameliorated tau hyperphosphorylation and the spatial memory deficits induced by PKA activation. We measured the total and activity-dependent Ser9-phosphorylated levels of glycogen synthase kinase (GSK)-3ß in hippocampal extracts. No significant change in the total level of GSK-3ß was observed between the different groups. However, compared with forskolin injection alone, pretreatment with escitalopram increased the level of Ser9-phosphorylated GSK-3ß. We also demonstrated that escitalopram increased Akt phosphorylation at Ser473 (the active form of Akt). Furthermore, we identified other important kinases and phosphatases, such as protein phosphatase 2A, extracellular signal-regulated kinases 1 and 2, and MAP kinase kinase-1/2, that have previously been reported to play a crucial role in tau phosphorylation; however, we did not detect any significant change in the activation of these kinases or phosphatases in our study. We unexpectedly demonstrated that forskolin caused anxiety-like behavior in rats, and pretreatment with escitalopram did not significantly ameliorate the anxiety-like behavior induced by forskolin. These data provide the first evidence that escitalopram ameliorates forskolin-induced tau hyperphosphorylation and spatial memory impairment in rats; these effects do not occur via the anti-anxiety activity of escitalopram but may involve the Akt/GSK-3ß signaling pathway.

Escitalopram Ameliorates Forskolin-Induced Tau Hyperphosphorylation in HEK239/tau441 Cells.

To investigate the effect of escitalopram (a widely used and highly efficacious antidepressant from the SSRI class) on tau hyperphosphorylation, HEK293/tau441 cells were pretreated with 4 µM of forskolin for 2 h. Then we treated the cells with different doses of escitalopram (0, 5, 10, 20, 40, 80 µM) for 22 h. We measured the phosphorylation level of tau by Western blotting. It was shown that escitalopram could protect tau from hyperphosphorylation induced by pharmacological activation of protein kinase A (PKA) at a dose of 20, 40, and 80 µM in vitro. Interestingly, the same dose of escitalopram could also increase the level of serine-9-phosphorylated GSK-3ß (inactive form) and the phosphorylation level of Akt at Ser473 (active form) with no significant change in the level of total GSK-3ß and Akt. Unexpectedly, 5-hydroxytryptamine 1A receptor (5-HT1A) agonist 8-OH-DPAT did not decrease forskolin-induced tau hyperphosphorylation. Our results suggest that escitalopram can ameliorate forskolin-induced tau hyperphosphorylation, which is not through the typical 5-HT1A pathway, and Akt/GSK-3ß signaling pathway is involved. These findings may support an effective role of antidepressants in the prevention of dementia associated with depression in patients.

Protein kinase Mζ is involved in the modulatory effect of fluoxetine on hippocampal neurogenesis in vitro.

The efficacy of chronic selective serotonin reuptake inhibitors (SSRIs) on depression is paralleled by the recovery of deficits in hippocampal neurogenesis related to sustained stress and elevated glucocorticoids. Previous studies have shown that atypical protein kinase C (aPKC) is implicated in the regulation of neurogenesis and the antidepressant response. Whether the specific aPKC isoforms (PKCζ, PKMζ and PKCι) are involved in SSRI-induced hippocampal neurogenesis and the underlying mechanisms is unknown. The present study shows that PKMζ and PKCι but not PKCζ are expressed in rat embryonic hippocampal neural stem cells (NSCs), whereas PKMζ but not PKCι expression is increased by the SSRI fluoxetine both in the absence and presence of the glucocorticoid receptor agonist dexamethasone. PKMζ shRNA significantly decreased neuronal proliferation and neuron-oriented differentiation, increased NSC apoptosis, and blocked the stimulatory effect of fluoxetine on NSC neurogenesis. Fluoxetine significantly increased PKMζ expression in hippocampal NSCs in a 5-hydroxytryptamine-1A (5-HT1A) receptor-dependent manner in both the absence and presence of dexamethasone. The PKMζ peptide blocker ZIP and MEK inhibitor U0126 significantly inhibited the increase in extracellular signal-regulated kinase 1/2 and cyclic adenosine monophosphate response element binding protein phosphorylation in the mitogen-activated protein kinase (MAPK) pathway and hippocampal NSC neurogenesis in response to fluoxetine and the 5-HT1A receptor agonist 8-OH DPAT. Collectively, our results suggest that the SSRI fluoxetine increases hippocampal NSC neurogenesis via a PKMζ-mediated mechanism that links 5-HT1A receptor activation with the phosphorylation of the downstream MAPK signaling pathway.

[Cloning, expression and immunodiagnostic evaluation of enolase from Echinococcus granulosus].

OBJECTIVE: To clone and express EgEno gene of Echinococcus granulosus, and to investigate the immunogenicity and diagnostic value of recombinant EgEno. METHODS: Total RNAS of E. granulosus was extracted and reversedly transcripted to cDNA. EgEno gene was amplified from cDNA and inserted into vector pET28a. The recombinant plasmid pET28a-EgEno was transformed into E. coli BL21 (DE3) for expression under the induction of IPTG. The expressed product was identified by SDS-PAGE and Western blotting. The purified recombinant EgEno protein was detected by ELISA with the sera of cystic echinococcosis patients, healthy persons and other patients. RESULTS: The EgEno gene was successfully amplified from cDNA of E. granulosus and a fusion protein was expressed in E. coli BL21 (DE3). The molecular weight of the expressed protein was around 50 kDa. The result of Western blotting indicated that the antigenicity of the protein was specific. The sensitivity of diagnosis by ELISA for cystic echinococcosis was 81.25%. CONCLUSION: EgEno of E. granulosus is cloned and expressed in E. coli BL21 (DE3) successfully, which might be used as a candidate antigen of immunodiagnosis for cystic echinococcosis.