Effect of Culture Supernatant Derived from Trichophyton Rubrum Grown in the Nail Medium on the Innate Immunity-related Molecules of HaCaT / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Trichophyton rubrum is superficial fungi characteristically confined to dead keratinized tissues. These observations suggest that the soluble components released by the fungus could influence the host immune response in a cell in contact-free manner. Therefore, this research aimed to analyze whether the culture supernatant derived from T. rubrum grown in the nail medium could elicit the immune response of keratinocyte effectively.</p><p><b>METHODS</b>The culture supernatants of two strains (T1a, T XHB ) were compared for the β-glucan concentrations and their capacity to impact the innate immunity of keratinocytes. The β-glucan concentrations in the supernatants were determined with the fungal G-test kit and protein concentrations with bicinchoninic acid protein quantitative method, then HaCaT was stimulated with different concentrations of culture supernatants by adopting morphological method to select a suitable dosage. Expressions of host defense genes were assessed by quantitative polymerase chain reaction after the HaCaT was stimulated with the culture supernatants. Data were analyzed with one-way analysis of variance, followed by the least significant difference test.</p><p><b>RESULTS</b>The T. rubrum strains (T1a and T XHB ) released β-glucan of 87.530 ± 37.581 pg/ml and 15.747 ± 6.453 pg/ml, respectively into the media. The messenger RNA (mRNA) expressions of toll-like receptor-2 (TLR2), TLR4, and CARD9 were moderately up-regulated in HaCaT within 6-h applications of both supernatants. HaCaT cells were more responsive to T1a than T XHB . The slight increase of dendritic cells-specific intercellular adhesion molecule 3-grabbing nonintegrin expression was faster and stronger, induced by T1a supernatant than T XHB . The moderate decreases of RNase 7, the slight up-regulations of Dectin-1 and interleukin-8 at the mRNA level were detected only in response to T1a rather than T XHB . After a long-time contact, all the elevated defense genes decreased after 24 h.</p><p><b>CONCLUSION</b>The culture supernatant of T. rubrum could directly and transiently activate the innate immune response of keratinocytes.</p>

Exposure to heat-inactivated Trichophyton rubrum resulting in a limited immune response of human keratinocytes / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Trichophyton rubrum (T. rubrum) represents the most important agent of dermatophytosis in humans. T. rubrum infection causes slight inflammation, and tends to be chronic and recurrent. It is suggested that it may result from the failure of epithelial cells to recognize T. rubrum effectively and initiate effective immune responses. The C-type lectin receptors (CLR) and toll-like receptors (TLR) are the two major pattern recognition receptors (PRRs) that recognize fungal components. Therefore, the purpose of the study was to analyze the expression of those PRRs and the cytokines in HaCaT cells stimulated with heat-inactivated T. rubrum conidia and hyphae, respectively.</p><p><b>METHODS</b>HaCaT cells were unstimulated or stimulated with heat-inactivated T. rubrum conidia and hyphae (1×10(6) and 1.5×10(5) colony-forming unit (CFU) in 2 ml medium, respectively) for 6, 12 and 24 hours. The mRNA expression of PRRs involved in recognizing fungal pathogen-associated molecular patterns (PAMPs) and signaling molecules were measured by quantitative reverse transcription polymerase chain reaction (RT-PCR). Meanwhile, surface toll-like receptor (TLR) 2, TLR4 and Dectin-1 were analyzed by fluorescence-activated cell sorter (FACS) 24 hours after treatment. The cytokines were detected in cell culture supernatants of HaCaT cells in 12 and 24 hours after treatment.</p><p><b>RESULTS</b>HaCaT cells constitutively expressed mRNA of membrane-bound TLR1, 2, 4 and 6, Dectin1 and DC-SIGN, but not Dectin-2 or Mincle. Heat-killed T. rubrum did not significantly upregulate gene transcriptions of the PRRs of HaCaT cells. Heat-inactivated T. rubrum conidia significantly reduced the surface expression of TLR2 and Dectin-1, and suppressed the secretions of interferon-inducible protein-10 (IP-10) and monocyte chemotactic protein-1 (MCP-1) of HaCaT cells, while heat-killed T. rubrum hyphae significantly induced the secretions of IP-10 and MCP-1.</p><p><b>CONCLUSION</b>The cell-wall antigens of T. rubrum fail to activate transcriptional expression of PRRs and induce a lower immune response of HaCaT cells by limited cytokines secretion.</p>

Effective reinnervation of the quadriceps femoris by spinal ventral root cross-anastomosis in rats / Reinervação efetiva do quadríceps femoral pela anastomose cruzada via raiz espinhal ventral em ratos

PURPOSE: To study the effective recovery of the quadriceps femoris by spinal ventral root cross-anastomosis in rats. METHODS: End-to-end anastomosis was performed between the left L1 and L3 ventral roots using autogenous nerve graft ,and the right L1 and L3 roots were left intact. In control animals, the left L3 ventral root was cut and shortened, and anastomosis was not performed. Six months postoperatively, the movement of low extremities was detected by electrophysiological examination, hindlimb locomotion and basso, beattie and bresnahan (BBB) scoring at one, three, seven, 14, 21 and 28 days after SCI. Fluorescence retrograde tracing with TRUE BLUE (TB) and HE staining were performed to observe the nerve regeneration. RESULTS: Six months after surgery, the anastomotic nerve was smooth and not atrophic. The amplitudes of action potential were 7.63±1.86 mV and 6.0±1.92 mV respectively before and after the spinal cord hemisection. The contraction of left quadriceps femoris was induced by a single stimulation of the anastomotic nerve. The locomotion of left hindlimb was partially restored after spinal cord hemisection while creeping and climbing. In addition, there was significant difference in the BBB score at one, three and seven days after SCI. TB retrograde tracing and neurophysiologic observation indicated efficient reinnervation of the quadriceps femoris. CONCLUSION: The cross-anastomosis between spinal ventral root can partially reconstruct the function of quadriceps femoris following SCI and may have clinical implication for the treatment of human SCI.

OBJETIVO: Investigar a recuperação efetiva do músculo quadríceps femoral pela anastomose cruzada via raiz espinhal ventral em ratos. MÉTODOS: Anastomose término-terminal foi realizada entre as raízes ventrais L1 e L3 à esquerda usando enxerto autógeno de nervo e, à direita, as raízes L1 e L3 foram mantidas intactas. Nos animais controles, à esquerda, a raiz ventral de L3 foi cortada e encurtada sem realização de anastomose. Após seis meses, o movimento das extremidades posteriores foi estudado por exame eletrofisiológico, e pelo escore de basso, beattie e bresnahan (BBB) com um, três, sete, 14, 21 e 28 dias após SCI. Fluorescência retrograde feita com TRUE BLUE (TB) e coloração com HE foram realizadas para observar a regeneração do nervo. RESULTADOS: Seis meses após a cirurgia, a anastomose do nervo estava lisa e sem atrofia. As amplitudes dos potenciais de ação foram 7,63±1,86 mV e 6,0±1,92 mV respectivamente antes e após a hemisecção da medula espinhal. A contração do músculo quadríceps femoral foi induzida por um único estímulo do nervo anastomosado. A locomoção do membro posterior esquerdo foi parcialmente restaurada após hemisecção da medula espinhal ao rastejar e escalar. Ademais, houve diferença significante no escore BBB nos dias um, três e sete após SCI. O traçado da TB retrógrada e a observação neurofisiológica indicaram reinervação eficiente do quadríceps femoral. CONCLUSÃO: A anastomose cruzada entre as raízes espinhais ventrais podem reconstruir parcialmente a função do quadríceps femoral após SCI e pode ter implicação clínica para o tratamento da SCI.

Phloroglucinol: safe and effective for the prevention of bladder spasm after TURP / 中华男科学杂志

<p><b>OBJECTIVE</b>To evaluate the efficacy of phloroglucinol in preventing bladder spasm after transurethral resection of the prostate (TURP).</p><p><b>METHODS</b>Using the random sampling method, we assigned 74 cases of TURP into a treatment group (n = 39), given 80 mg phloroglucinol every day for 3 days, and a control group (n = 35), left untreated. Then we observed the frequency, duration and pain of bladder spasm within the 3 days and compared them between the two groups.</p><p><b>RESULTS</b>The mean frequency, duration and pain visual analogue score of bladder spasm were (4.3 +/- 1.2) times, (7.2 +/- 2.1) min and 3.2 +/- 1.6 respectively in the treatment group, as compared with (7.5 +/- 2.4) times, (15.6 +/- 6.8) min and 4.7 +/- 2.3 in the control, with statistically significant differences between the two groups (P < 0.05). And no obvious adverse reactions were found in the treatment group.</p><p><b>CONCLUSION</b>Phloroglucinol is safe and effective for the prevention and treatment of bladder spasm following TURP.</p>

Effect of cedemex on cAMP and cGMP levels of different brain areas in morphine withdrawal rats / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the effect of Cedemex on cAMP and cGMP contents in different brain regions in morphine withdrawal rats precipitated by naloxone.</p><p><b>METHOD</b>A physical morphine dependent model of rats was established by subcutaneous injection of morphine in gradually increasing dosage within 7 days. cAMP and cGMP contents of VTA, cortex and hippocampus of the rat brains were determined by radioimmunoassay.</p><p><b>RESULT</b>The morphine withdrawal symptoms of rats were relieved significantly by ig Cedemex. Compared with the controls, cAMP content in the region of VTA, cortex and hippocampus of the morphine dependent rats were significantly higher (P < 0.05), while cGMP contents in those regions were significantly lower (P < 0.05). cAMP contents in the area of VTA, cortex and hippocampus of the morphine dependent rats were significantly reduced, while cGMP contents were significantly increased by ig Cedemex.</p><p><b>CONCLUSION</b>Cedemex may significantly attenuate the morphine withdrawal symptoms in rats. The mechanism of this effect may be related to adjusting the contents of cAMP and cGMP in some brain regions.</p>

Effect of manganese on apoptosis in striatum neurons of rats / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To explore the mechanism of neurotoxicity induced by manganese, and observe the effects on the apoptosis of neurons in rat striatum.</p><p><b>METHODS</b>SD rats were divided into four groups, six rats each group. Three dose groups were exposed to high, middle, and low level of MnCl(2). At the end of experiment, all rats of the exposed groups and control group were decapitated, their striatums were removed and the Mn content of striatum, the apoptotic morphology, ratio and ultrastructural organization were analyzed.</p><p><b>RESULTS</b>The Mn content of striatum and apoptosis index of the three dose groups exposed to high, middle, and low level of Mn were significantly higher than control group (P < 0.05). The Mn content of striatum of the three dose groups exposed to high, middle, low level of MnCl(2) and control group were 2.98 +/- 0.52, 2.75 +/- 0.37, 2.61 +/- 0.73, 0.60 +/- 0.20 respectively. The apoptosis index of striatum of the three dose groups exposed to high, middle, low level of MnCl(2) and control group were 24.83 +/- 5.98, 17.00 +/- 5.33, 15.33 +/- 2.58, 2.83 +/- 0.41 respectively, and following higher level dose, the apoptosis index increased. The nucleus of neurons in striatum become smaller, condensed, etc, and these character showed apoptosis of neurons.</p><p><b>CONCLUSION</b>Mn can result in apoptotic morphology and increase level of apoptosis in striatum. The level of apoptos varies with Mn concentration.</p>

Effect of manganese on cytosolic free calcium concentration in cortical neurons / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To investigate the change of free Ca(2+) in cytoplasma in the neurotoxicity of the manganese (Mn).</p><p><b>METHODS</b>The cortical neurons were separated from the neonatal Wistar rats and cultured in vitro. The neurons were grouped as the Mn-treated groups and the untreated group. The neurons in the Mn-added groups were incubated in the culture media containing lower, medium and high dosage manganese chloride (MnCl(2 x 4) H2O) with the concentration at 0.2, 0.6, 1.0 mmol/L respectively. Meanwhile, neurons in control were cultured in the normal culture media. All treatments stopped 24 h later. Neurons were labeled Ca(2+) sensitive prober, Fluo-3/AM. The fluorescence intensity of Fluo-3 combined with Ca(2+) was examined by LSCM (Laser scanning confocal microscope) and was treated by the picture analysis technique. The intensity was equal to the free Ca(2+) concentrations in cytoplasma of neurons.</p><p><b>RESULTS</b>MnCl(2) can induce free Ca(2+) overloaded in cytoplasma of neurons, but the increasing degree varied in MnCl(2) dosage. Cytoplasma Ca(2+) concentration in the moderate dosage The moderate dosage MnCl(2) group and the high dosage MnCl(2) group were significantly higher than that in the lower dosage MnCl(2) group and the control group (P < 0.05).</p><p><b>CONCLUSION</b>The Ca(2+) overload is involved in the neurotoxicity of manganese, and a dosage response relationship is found between the manganese chloride dose and Ca(2+) overload in cortical neurons.</p>