Impaired CBS-H2S signaling axis contributes to MPTP-induced neurodegeneration in a mouse model of Parkinson's disease.

Hydrogen sulfide (H2S), a novel neuromodulator, is linked to the pathogenesis of several neurodegenerative disorders. Exogenous application of H2S exerts neuroprotection via anti-inflammation and anti-oxidative stress in animal and cellular models of Parkinson's disease (PD). However, the role of endogenous H2S and the contribution of its various synthases in PD remain unclear. In the present study, we found a decline of plasma and striatal sulfide level in 1-methy-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) induced PD mouse model. Interestingly, among the three H2S generating enzymes, only cystathionine ß-synthase (CBS) expression was largely reduced in the striatum of MPTP-treated mice. The in vitro study confirmed a significant decrease of CBS expression in 1-methyl-4-phenylpyridinium (MPP+)-stimulated astrocytes and microglia, but not in neurons or SH-SY5Y dopaminergic cells. Striatal CBS overexpression, elicited by stereotaxic delivery with Cbs gene using recombinant adeno-associated-virus (rAAV-Cbs), successfully enhanced the sulfide level in the striatum and partially rescued the MPTP-induced dopaminergic neurotoxicity in the midbrain. Specifically, striatal CBS overexpression alleviated the motor deficits and dopaminergic neuron losses in the nigro-striatal pathway, with a concomitant inhibition of glial activation in MPTP-treated mice. Furthermore, compared to rAAV-Vector, rAAV-Cbs injection reduced the aberrant accumulation of nitric oxide and 3-nitrotyrosine (an indicator of protein nitration) in the striatum of MPTP-treated mice. Notably, it also attenuated the increase of nitrated α-synuclein level in MPTP mice. The in vitro study demonstrated that lentivirus-mediated CBS overexpression elevated the sulfide generation in glial cells. Moreover, glial CBS overexpression offered protection to midbrain dopaminergic neurons through repressing nitric oxide overproduction in both glial and neuronal cells induced by MPP+. Taken together, our data suggest that impaired CBS-H2S axis may contribute to the pathogenesis of PD, and that modulation of this axis may become a novel therapeutic approach for PD.

Urate promotes SNCA/α-synuclein clearance via regulating mTOR-dependent macroautophagy.

Serum urate levels are reported to be significantly lowered in patients with Parkinson's disease (PD) and inversely correlated to the risk and progression of PD. However, the mechanism by which urate affects PD is poorly understood. Here we showed that treatment with uric acid (UA) resulted in an autophagy activity enhancement in PC12 cells in dose- and time-dependent manners, as indicated by LC3-II increase and P62 decrease. Moreover, UA was still able to increase the LC3-II level and the number of LC3 puncta in the presence of Bafilomycin A1, a lysosomal inhibitor. These changes of autophagic markers were preceded by mTOR inhibition and ULK1 activation. Co-treatment with 3-benzyl-5-((2-nitrophenoxy) methyl)-dihydrofuran-2(3H)-one (3BDO), an mTOR activator, abolished the UA-induced LC3-II increase. More importantly, UA reduced SNCA/α-synuclein accumulation in PC12 cells that overexpress wildtype or A53T mutant SNCA, and this was blocked by Bafilomycin A1 co-treatment. The in vivo study showed that UA administration was able to modulate the levels of autophagy markers, increase the autophagosome/autolysosome formation, and reduce SNCA accumulation in the midbrain of SNCAA53T transgenic mice. Taken together, our findings suggest that UA could induce autophagy activation via an mTOR-dependent signaling and ameliorate SNCA accumulation. This implicates that urate-elevating agent may become a potential strategy for PD therapy.

The role of follicular T helper cells in patients with malignant lymphoid disease.

OBJECTIVES: To investigate the dynamic change of follicular T helper cells (TFH) in patients with malignant lymphoid disease (MLD) and to explore its clinical significance. METHODS: The dynamic change of TFH cells, ICOS+- and PD-1+ TFH cells at pretreatment and different treatment periods was determined by flow cytometry in 85 MLD patients. Concentration of interleukin 21 (IL-21) was evaluated by ELISA, and the correlation between clinical prognosis and the ratio of TFH cells was analyzed. RESULTS: Significantly increased ICOS+- and PD-1+ TFH cells were found in MLD patients at pretreatment compared to healthy controls. Decreased or even close to normal levels of ICOS+- and PD-1+ TFH cells were found at the end of treatment. However, in the patients with progressive disease, high levels of ICOS+- and PD-1+ TFH cells were found. Moreover, a significantly increased plasma IL-21 level was found in MLD patients. Negative correlation was found between the level of ICOS+-, PD-1+ TFH cells, as well as IL-21 and the prognosis of MLD. CONCLUSIONS: Significantly increased TFH cell ratios were found in patients with MLD, and decreased TFH cells ratios could be expected in those treatment-effective patients, which could be used as the therapeutic efficacy index.

Alteration of gene expression by zinc oxide nanoparticles or zinc sulfate in vivo and comparison with in vitro data: A harmonious case.

Granulosa cells (GCs) are those somatic cells closest to the female germ cell. GCs play a vital role in oocyte growth and development, and the oocyte is necessary for multiplication of a species. Zinc oxide (ZnO) nanoparticles (NPs) readily cross biologic barriers to be absorbed into biologic systems that make them promising candidates as food additives. The objective of the present investigation was to explore the impact of intact NPs on gene expression and the functional classification of altered genes in hen GCs in vivo, to compare the data from in vivo and in vitro studies, and finally to point out the adverse effects of ZnO NPs on the reproductive system. After a 24-week treatment, hen GCs were isolated and gene expression was quantified. Intact NPs were found in the ovary and other organs. Zn levels were similar in ZnO-NP-100 mg/kg- and ZnSO4-100 mg/kg-treated hen ovaries. ZnO-NP-100 mg/kg and ZnSO4-100 mg/kg regulated the expression of the same sets of genes, and they also altered the expression of different sets of genes individually. The number of genes altered by the ZnO-NP-100 mg/kg and ZnSO4-100 mg/kg treatments was different. Gene Ontology (GO) functional analysis reported that different results for the two treatments and, in Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment, 12 pathways (out of the top 20 pathways) in each treatment were different. These results suggested that intact NPs and Zn(2+) had different effects on gene expression in GCs in vivo. In our recent publication, we noted that intact NPs and Zn(2+) differentially altered gene expression in GCs in vitro. However, GO functional classification and KEGG pathway enrichment analyses revealed close similarities for the changed genes in vivo and in vitro after ZnO NP treatment. Furthermore, close similarities were observed for the changed genes after ZnSO4 treatments in vivo and in vitro by GO functional classification and KEGG pathway enrichment analyses. Therefore, the effects of ZnO NPs on gene expression in vitro might represent their effects on gene expression in vivo. The results from this study and our earlier studies support previous findings indicating ZnO NPs promote adverse effects on organisms. Therefore, precautions should be taken when ZnO NPs are used as diet additives for hens because they might cause reproductive issues.

microRNA-7 Protects Against 1-Methyl-4-Phenylpyridinium Iodide-Induced Cell Apoptosis in SH-SY5Y Cells by Directly Targeting Krüpple-Like Factor 4.

This study intended to investigate the role and underling mechanism of microRNA-7 (miR-7) on neuronal death in Parkinson's disease (PD). Human neuroblastoma cell line SH-SY5Y was employed and 1-methyl-4-phenylpyridinium iodide [MPP(+)] was used to generate PD model in vitro. Furthermore, an upregulation of miR-7 was performed in SH-SY5Y by transfection with miR-7 mimics. Cell viability and cell apoptosis were determined. Moreover, the target and the mechanism of miR-7 in MPP(+)-induced cell death were also investigated. The upregulation of miR-7 promoted cell viability and suppressed cell apoptosis in MPP(+)-treated SH-SY5Y cells. Furthermore, miR-7 could directly bind to the 3'-untranslated region of Krüppel-like factor 4 (KLF4, positions 574-580). Moreover, knockdown of KLF4 by the specific siRNA inhibited SH-SY5Y apoptosis under MPP(+) treatment. In addition, KLF4 overexpression apparently attenuated the protective effect of miR-7 in MPP(+)-induced SH-SY5Y apoptosis. This study indicated that miR-7 protects from MPP(+)-induced cell apoptosis in SH-SY5Y by directly targeting KLF4.

[Sensitivity to bortezomib of RPIM8226 cells after co-cultured with down-regulated Cav-1 expression HUVECs].

OBJECTIVE: To investigate the sensitivity to bortezomib of RPMI8226 cells after co-cultured with down-regulated Caveolin (Cav)-1 expression of HUVECs by transfection with Cav-1 shRNA (HUVECs(Cav-1 low)). METHODS: Exposure to bortezomib with or without 50 nmol/L dexamethasone at different concentration, the proliferation of RPMI8226 was analyzed by MTT assay when it was cultured alone or co-cultured with HUVECs(Cav-1 low). Cav-1 expression was detected by using of Western blot and cell cycle, apoptosis and the level of reactive oxygen species (ROS) were analyzed by flow cytometry. RESULTS: Cav-1 expression was notably down-regulated in HUVECs(Cav-1 low) (0.2199±0.0288 vs 1.3195±0.2393) (P<0.01). The IC(50) of bortezomib for RPMI8226 cultured alone, co-cultured with HUVECs orHUVECCav- 1 low were 20 nmol/L, 50 nmol/L and 65 nmol/L, respectively. The percentages of G0/G1 phase in RPMI8226 cultured alone, co-cultured with HUVECs and HUVECs(Cav-1 low) were 28.49%, 30.41%, and 36.15% respectively. The protection of RPMI 8226 against apoptosis by HUVECs was demonstrated that the apoptosis/death rates were 66.8%, 10.7% and 8.6% in RPMI8226 cultured alone, co-cultured with HUVECs and HUVECs(Cav-1 low) after exposure to 20 nmol/L bortezomib for 24 h. RPMI8226 could induce the oxidative stress of HUVECs before and after co-culture. The ROS level was raised from 15.0% to 35.2% in RPMI8226, from 80.4% to 91.0% in HUVECs, and from 84.6% to 96.8% in HUVECs(Cav-1 low). CONCLUSION: The down-regulated Cav-1 expression of HUVECs could promote proliferation and induce apoptosis of RMPI8226 cells, lead to G0/G1 phase arrest, and reduce the sensitivity to bortezomib.

A meningioma with peripheral rim enhancement on MRI.

Meningiomas are common, typically benign intracranial neoplasms with well-demarcated borders.Meningiomas with indistinct boundaries have been reported.These can invade surrounding structures, and present surgical and diagnostic challenges. We present the case of an unusual meningioma in a 53-year-old male in which preoperative magnetic resonance imaging (MRI) revealed an irregular lesion with clear boundaries and peripheral rim enhancement. Intraoperatively, however, no cleavage plane was apparent. Histological examination showed an increase of fibroconnective tissue with proliferation of dilated vessels in the periphery of the tumor concordant with the rim.Immunohistochemical staining of the tumor was positive for EMA and CD34, but negative for CEA, Ki67, and GFAP. Immunohistochemical staining of proliferating vessels in the periphery of the tumor was positive for CD34. A so-called 'capsule' structure was suggested according to MRI findings and pathological examination.The tumor was diagnosed as a mixed type meningioma,WHO grade I.

[Influence of iron ion on the growth of Trichomonas vaginalis in vitro].

OBJECTIVE: To study the influence of iron ion on the growth of Trichomonas vaginalis in vitro. METHODS: Quantitative pure incubation of T. vaginalis trophozoites with an initial density of 1 x l0(5)/ml was carried out at 37 degrees C in TYM (trypticase-yeast extract-maltose) medium (pH 6.0) with additional 100, 200, 300, and 400 micromol/L iron ion, respectively, and in TYM medium without additional iron ion as control The numbers of live and dead trichomonads were counted under an optical microscope after trypan blue staining, and the growth curve and survival rate of T. vaginalis were drawn to calculate the generation time of T. vaginalis. The minimal lethal concentration (MLC) of metronidazole was tested by serial dilution method when the trophozoites of T. vaginalis were incubated in TYM medium with 200 micromol/L iron ion and in control group without additional iron ion. RESULTS: The maximum densities of 2.9 x 10(6), 3.2 x 10(6), 3.1 x 10(6), and 2.8 x 10(6)/ml were obtained when the trophozoites were incubated for 40 h in TYM medium with 100, 200, 300, and 400 micromol/L iron ion, respectively, while the maximum density was achieved at 50 h in the control, which was 2.5 x l0(6)/ml. The generation time was (6.8 +/- 0.7) h in 400 micromol/L iron ion group, longer than those in the groups with 100-300 micromol/L iron ion, which were (4.8 +/- 0.3), (4.8 +/- 0.2), and (5.0 +/- 0.4) h, respectively (P < 0.05). While the generation times in 100-400 micromol/L groups were all shorter than that of the control [(10.2 +/- 3.1) h] (P < 0.05). MLC of metronidazole for T. vaginalis in TYM medium with 200 micromol/L iron ion was (23.44 +/- 11.56) microg/ml, significantly lower than that of the control [(31.25 +/- 15.44) microg/ml] (P < 0.05). CONCLUSION: In TYM medium with 100-400 micromol/L iron ion, T. vaginalis trophozoites reproduce faster, and the MLC of metronidazole to the parasites is lower.

[Cryopreservation of Trichomonas vaginalis and its influence factors].

Different concentrations of dimethylsulfoxide (DMSO) and glycerol, various densities of Trichomonas vaginalis trophozoites and duration of storage were used to study the effects on the cryopreservation of T. vaginalis at -78 degrees C. The optimal densities of trichomonads for cryopreservation were (1-2) x10(6)/ml. Glycerol and DMSO showed a high protective effect, with a maximum effect at a concentration of 10%, and the survival rates were 38.0% and 31.7%, respectively. There was no significant difference between them (P > 0.05). The results showed that the above optimized factors can provide a good protection in short-term (1-16 weeks) cryopreservation, and the trichomonads can be incubated to a logarithmic growth phase at 37 degrees C.

Organocatalytic peroxy-asymmetric allylic alkylation.

The peroxy-asymmetric allylic alkylation of hydroperoxyalkanes with Morita-Baylis-Hillman carbonates was catalysed by modified cinchona alkaloids (up to 93% ee), from which chiral alpha-methylene-beta-hydroxy esters could be efficiently derived.

[The reperfusion injury model improvement and the tolerance time investigation of rabbit spinal cord ischemia under normothermia].

OBJECTIVE: This study is designed to improve the rabbit model of ischemic- reperfusion injury and determine the safe clamping duration relevant to the spinal cord tolerance to ischemia at normothermia. METHODS: 50 New Zealand white rabbits were assigned randomly to 5 groups (Group C20, C25, C30, C40 and C60, 10 rabbits in each group) according to different clamping durations, ranging from 20 min to 60 min. The rabbits were endotracheally intubated for ventilation, and their left ear arteries were catheterized for monitoring the mean artery pressure. The spinal cord ischemia was induced by infrarenal aorta occlusion. A catheter was inserted into the aorta distal clamped site for monitoring the distal artery pressure. The neurological functional status of animal was assessed with the Tarlov scale system (0 or 1 meaning the rabbit paraplegia), at the moment of revival, 6 h, 24 h, and 48 h after the reperfusion. After last scoring, the lumbar segments of spinal cord (L4-L6) were removed for pathological examination, and the normal motor neurons of anterior horn were counted. RESULTS: Forty-eight hours after the infusion, the severe neurological impairments were not detected in the rabbits whose aorta were only clamped for 20 min (Group C20). However, the rabbits in Group CSO became totally paraplegic, and the rabbits in Group C25 C30 or C40 developed the paraplegia at 30% , 80% or 90% respectively. The median number of normal motor neuron was 12. 5, 10 or 2 respectively in Group C20, C25 or C30, and 0 median number resulted in Group C40 and C60. CONCLUSION: The rabbit model of ischemic-reperfusion injury is successfully improved, of which the safe clamping duration without spinal cord injury is not more than 20 min at normothermia.