Determination of Th9 cells and IL-9 in children with Mycoplasma pneumoniae infection / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the clinical significance of T helper type 9 (Th9) cells and interleukin-9 (IL-9) in children suffering from Mycoplasma pneumoniae (MP) infection.</p><p><b>METHODS</b>A total of 86 children who were diagnosed with MP infection between January 2013 and June 2014 were classified into upper respiratory infection (URI) group (n=29), mild MP pneumonia (MPP) group (n=32) and severe MPP group (n=25). Twenty-eight healthy children were used as the control group. Peripheral blood mononuclear cells were isolated by Ficoll density gradient centrifugation, and the percentage of Th9 cells in peripheral blood was measured by flow cytometry. Serum IL-9 level was determined using ELISA.</p><p><b>RESULTS</b>The URI, mild MPP, and severe MPP groups had significantly higher percentages of Th9 cells and IL-9 levels than the control group (P<0.05); the mild MPP and severe MPP groups had significantly higher percentages of Th9 cells and IL-9 levels than the URI group (P<0.05), and the two indices were significantly higher in the severe MPP group than in the mild MPP group (P<0.01).</p><p><b>CONCLUSIONS</b>Children with MP infection have an elevated percentage of Th9 cells and IL-9 expression, both of which are positively correlated with the severity of the disease. It can be predicted that Th9 cells and IL-9 can be used as evaluation indicators for the progression and outcome of children with MP infection.</p>

Effects of safflower injection on HEL leukemia cell proliferation and apoptosis and relevant molecular mechanisms / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the effect of safflower injection on the proliferation and apoptosis of human leukemia cell line HEL and the relevant molecular mechanisms.</p><p><b>METHODS</b>HEL cells were treated with different concentrations of safflower injection. HEL cells without safflower injection treatment were used as the control group. MTT method was used to detect the inhibitory rate of the HEL cells at 24, 48 and 72 hours after various concentrations of safflower injection treatment (10, 20, 30, 40 and 50 mg/mL). The cell cycle and apoptosis were detected using flow cytometry and the HOXB3-mRNA expression was measured by RT-PCR at 48 hours after safflower injection treatment (10, 20 and 30 mg/mL).</p><p><b>RESULTS</b>Compared with the control group, various concentrations of safflower injection inhibited HEL cell proliferation in a dose-dependent manner (P<0.05). At 48 hours after various concentrations of safflower injection treatment, the number of treated cells in the G2/M phase increased, but that in the S phase decreased, and the apoptosis rate was significantly higher than that in the control group, with a dose-dependent manner (P<0.05). The expression of HOXB3-mRNA in safflower injection-treated cells decreased in a dose-dependent manner compared with the control group (P<0.05).</p><p><b>CONCLUSIONS</b>Safflower injection can inhibit proliferation and induce apoptosis of HEL cells in vitro, and its underlying mechanisms may involve down-regulation of the HOXB3-mRNA expression.</p>

Epigallocatechin gallate attenuates the expression of regulated upon activation normal T cell expressed and secreted induced by lipopolysaccharide in human retinal endothelial cells / 生理学报

The present study was undertaken to determine the effect of epigallocatechin gallate (EGCG) on lipopolysaccharide (LPS)-induced production of inflammatory chemokine regulated upon activation normal T cell expressed and secreted (RANTES) in human retinal endothelial cells (HRECs) and to explore the underlying regulatory mechanism. HRECs were stimulated with LPS in the presence or absence of EGCG at various concentrations (100, 50, 25, 12.5, 6.25 μmol/L). The optimum concentration of drug was determined by a real-time cell-electronic sensing (RT-CES) system, and MTS chromatometry was used to detect the toxicity of LPS and EGCG on HRECs. RANTES production in the culture supernatant was measured by ELISA. The expression levels of Akt and phosphorylated Akt were examined by Western blot assay. The result showed that LPS markedly stimulated RANTES secretion from HRECs. EGCG treatment significantly suppressed LPS-induced RANTES secretion in a dose-dependent manner. Furthermore, EGCG exhibited a dose-dependent inhibitory effect on LPS-induced phosphorylation of Akt. Taken together, our data suggest that EGCG suppresses LPS-induced RANTES secretion, possibly via inhibiting Akt phosphorylation in HRECs.

MGB probe assay for rapid detection of mtDNA11778 mutation in the Chinese LHON patients by real-time PCR / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

<p><b>OBJECTIVE</b>Leber's hereditary optic neuropathy (LHON) is a maternally inherited degeneration of the optic nerve caused by point mutations of mitochondrial DNA (mtDNA). Many unsolved questions regarding the penetrance and pathophysiological mechanism of LHON demand efficient and reliable mutation testing. This study aims to develop a minor groove binder (MGB) probe assay for rapid detection of mtDNA11778 mutation and heteroplasmy in Chinese LHON patients by real-time polymerase chain reaction (PCR).</p><p><b>METHODS</b>Forty-eight patients suspected of having LHON and their maternal relatives underwent a molecular genetic evaluation, with 20 normal individuals as a control group at the same time. A real-time PCR involving two MGB probes was used to detect the mtDNA11778 mutation and heteroplasmy. A linear standard curve was obtained by pUCmLHONG and pUCmLHONA clones.</p><p><b>RESULTS</b>All 48 LHON patients and their maternal relatives were positive for mtDNA11778 mutation in our assay, 27 heteroplasmic and 21 homoplasmic. Eighteen cases did not show an occurrence of the disease, while 9 developed the disease among the 27 heteroplasmic mutation cases. Eleven did not show an occurrence of the disease, while 10 cases developed the disease among 21 homoplasmic mutation cases. There was a significant difference in the incidence between the heteroplasmic and the homoplasmic mutation types. The time needed for running a real-time PCR assay was only 80 min.</p><p><b>CONCLUSION</b>This real-time PCR assay is a rapid, reliable method for mtDNA mutation detection as well as heteroplasmy quantification. Detecting this ratio is very important for predicting phenotypic expression of unaffected carriers.</p>

Analysis on the effect of secondary mutations on Leber's hereditary optic neuropathy / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the effect of secondary mutations on Leber's hereditary optic neuropathy (LHON).</p><p><b>METHODS</b>Three primary and 24 secondary mutations were identified in 4 Chinese families which included male offspring.</p><p><b>RESULTS</b>All of the four pedigrees carried classic LHON mutations at nucleotide (nt) 11778, and did not carry any point of 24 secondary mutations. Nevertheless many polymorphic points were found in the nearby fragments of these pedigrees, such as 5178, 5108, 3705, 3721, 13734, etc.</p><p><b>CONCLUSION</b>Male offspring sequences should be analyzed in pedigrees with LHON to avoid the influence of familial inheritance characteristic which mitochondrial DNA polymorphism carried. Existence of the "repair genes" may affect the development of LHON.</p>

Effects of pioglitazone on MKP-1 and TSP-1 expression in early stages of diabetic retinopathy induced by streptozotocin / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To explore the effects of pioglitazone on MKP-1 and TSP-1 expression in the early stages of diabetic retinopathy induced by streptozotocin (STZ) and the relevant mechanism in it.</p><p><b>METHODS</b>Diabetic rats were induced by an intraperitoneal injection of STZ in SD rats. Thirty male SD rats were randomly divided into 3 groups: diabetes adding pioglitazone group (intragastric administration pioglitazone 20 mg x kg(-1) x d(-1)), diabetes adding BBS group and normal control group. The body weight and blood glucose were measured every two weeks. Eight weeks later, all rats were killed and the expression of TSP-1 and MKP-1 mRNA was quantified in retinal tissue by immunohistochemistry and reverse transcription-polymerase chain reaction (RT-PCR) respectively.</p><p><b>RESULT</b>TSP-1 and MKP-1 concentrations were significantly increased in the diabetic rats' retinal tissue compared to the control rats. Diabetes groups adding pioglitazone caused the upregulation of TSP-1 and MKP-1 expression in the retina among the three groups.</p><p><b>CONCLUSION</b>Pioglitazone treatment can significantly attenuate the evolutionary in the early stages of experimental diabetic retinopathy. Further studies should address the possible involvement of TSP-1 and MKP-1 in the correlational pathophysiology between pioglitazone and diabetic retinopathy.</p>

Experimental induction of posterior vitreous detachment in rabbits with hyaluronidase and perfluoroethane (C2F6) / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To evaluate the experimental induction of posterior vitreous detachment (PVD) by intravitreous injection of hyaluronidase and perfluoroethane (C(2)F(6)).</p><p><b>METHODS</b>Fifteen rabbits (30 eyes) were divided into 3 experimental groups,the contralateral eyes in same animals served as the controls. Eyes in group A and B were received two vitreous injections of 15 IU of hyaluronidase at an interval of 5 d. The eyes in group C and all control eyes were injected with balanced salt solution (BSS). Seven days after injection, the experimental eyes in group A and C were received 0.5 ml of Fifteen rabbits (30 eyes) were divided into 3 experimental groups, the contralateral eyes in same animals served as the controls. Eyes in group A and B were received two vitreous injections of 15 IU of hyaluronidase at an interval of 5 d. The eyes in group C and all control eyes were injected with balanced salt solution (BSS). Seven days after injection,the experimental eyes in group A and C were received 0.5 ml of C(2)F(6) injection. The ocular and retinal signs were examined for 8 following weeks and then killed for histological examination.</p><p><b>RESULT</b>Five eyes in group A (100.0%) showed complete separation of the vitreous cortex from the retina (PVD), three eyes in group B(60.0%) showed partial PVD, and no PVD was detected in group C and all control eyes. On electroretinogram no significant difference was found in amplitude and latency of a-(or b-) wave in both experimental and control eyes, between before and after experiments. No evidence of ocular or retinal toxicity was revealed by light or scanning electronic microscopy in all eyes.</p><p><b>CONCLUSION</b>Vitreous injection of hyaluronidase combined with perfluoroethane, as a safety method, can induce posterior vitreous detachment without mechanical vitrectomy.</p>