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Objective To apply the BestSeqTM new generation pathogenic gene detection technology to perform the genetic detec‐tion in the patients with progressive muscular dystrophy (PMD) validating its sensitivity and specificity .Methods The BestSeqTM new generation pathogenic gene detection technology was used to perform the gene sequencing in 2 cases of limb‐girdle muscular dystrophy(LGMD) and 6 cases of Dunchenne′s muscular dystrophy(DMD) ,and the found point mutations were confirmed by the Sanger sequencing method .Results This study completed the genetic detection in above 8 cases ,2 cases of large fragment deletion and 10 cases of micromutations were detected ,in which 8 micromutations were the new mutation discovered ffor the first time and verified by the Sanger sequencing .Conclusion The BestSeqTM new generation pathogenic gene detection technology greatly increa‐ses the detection efficiency by using the high density imbricate type probe and multiple tag technology ,and has the better clinical ap‐plication prospects .
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Objective To explore the effects of natural avocado oil and olive oil on the proliferation and differentiation of HaCaT cells. Methods MTT assay was performed to determine the optimal work concentration of avocado oil and olive oil. Cultured HaCaT cells were divided into 3 groups, i.e., avocado oil group treated with avocado oil of 3% (v/v), olive oil group treated with olive oil of 3% (v/v), and control group without any treatment. Immunocytochemistry and immuno-dot-blot method were used to detect the expressions of c-myc, mitogen-activated protein kinase ( MAPK ), nuclear factor ( NF)-κB, filaggrin, involucrin and keratin10 in HaCaT cells. Results As immunocytochemistry showed, the mean grey values (staining intensity) of c-myc,MAPK, and NF-κB in HaCaT cells were 131.4 ± 6.6,136.3 ± 4.5 and 134.3 ± 5.2 respectively in the avocado oil group, 121.1 ± 4.5, 107.9 ± 7.3 and 106.4 ± 5.4 respectively in the olive oil group, significantly higher than that in the control group (101.9 ± 8.9,91.4 ± 5.1 and 94.3 ± 7.0, respectively, all P＜ 0.05), and the avocado oil group was higher than the olive oil group in all the above parameters (all P ＜ 0.05). Increased expressions of filaggrin, involucrin and keratin 10 were observed in the avocado oil group and olive oil group compared with the control group (all P＜ 0.05), and in the olive oil group than in the avocado oil group (all P＜ 0.05).The mean grey values of these proteins obtained by immunocytochemisty were significantly correlated with those obtained by immuno-dot-blot method in avocado oil group (r = 0.94, P ＜ 0.01 ) and olive oil group (r=0.97, P ＜ 0.01 ). Conclusions Certain concentrations of avocado oil and olive oil can promote the proliferation and differentiation of HaCaT cells; avocado oil is more capable to accelerate their growth and proliferation, and olive oil to enhance their differentiation.
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Objective To explore the expression of Tiaml in clear cell renal cell carcinoma and analyze its correlations to pathology of disease and prognosis.Methods The expressions of Tiam1 protein in 107 specimens of human clear cell renal cell carcinoma and 20 specimens of normal renal tissues were detected by immunohistochemical staining and its clinical significance was then analyzed.Results The expression of Tiam1 protein was higher in renal cancers than in the adjacent normal tissues ( P ＜ 0.01 ).Tiam1 protein expression rates were 47.6％ and 72.7％ in Ⅰ - Ⅱ and Ⅲ - Ⅳ tumors,while 49.3％ and 76.5％ in T1 - T2 and T3 - T4 tumors,respectively ( P ＜ 0.01 ).Expression of Tiam1 protein was higher in lymph node positive renal carcinoma tissues than in lymph node negative renal carcinoma tissues ( 71.7％ versus 47.5％,P ＜ 0.05 ).The expression of Tiam1 in carcinoma tissues showed a positive relationship with tumor vascular invasion (81.3％ versus 48.0％,P ＜ 0.01 ).In patients followed-up 5 - 8 years,Kaplan-meier analysis and the log-rank test showed that the 5-year survival was significantly different between the group of lower and higher Tiaml expression groups ( 84.4％ versus 46.8％,P ＜ 0.05 ).Conclusions The expression of Tiaml protein was higher in human primary renal carcinoma than in normal renal tissues.The positive rate of Tiam1 protein expression was related to classification,TNM stage,lymph node metastasis and vascular invasion.The detection of the expression of Tiaml protein may be helpful in the diagnosis and prognosis of renal carcinoma.
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Objective To study the effect of modified Xiaochaihu Decoction on NF-kB and HSP70 of BRL cell induced by cisplatin. Methods BRL Cells were divided into control group, cisplatin group, high and low dose of modified Xiaochaihu decoction. Complete cell protein dot blot technique and immunohistochemisth analysis were appllied to detect the change of NF-kB and HSP70. Results There was a significant increase in NF-kB and decrease in HSP70 of BRL cell induced by cisplatin compared with control group. There was a significant decrease of NF-kB in modified Xiaochaihu decoction group compared with cisplatin group. There were increases of the expression of HSP70 in both high and low dose group, but only the high dose group demonstrated significant difference. Conclution The expression of NF-kB of BRL cell induced by cisplatin can be inhibited while the expression of HSP70 can be enhenced by modified Xiaochaihu decoction. It may be one of important mechanisms of relieving cell oxidize lesion and inhibiting apoptosis by modified Xiaochaihu decoction.
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Objective To compare the difference in quercetin against oxidative stress response in mouse and in NIH-3T3 cells before and after H2O2 treatment,to explore the underlying mechanism for the quercetin antioxidant.Methods The cultured NIH-3T3 cells were randomly divided into 4 groups: quercetin(Q) pre-protective group(Qb) firstly treated with quercetin for 24 h followed by incubation with H2O2 for 30 min;post-protective group(Qa) treated with H2O2 for 30 min followed by incubation with quercetin for 24 h;H2O2 group(H2O2) after exposure to H2O2 for 30 min,incubated with DMEM medium and the control group(C) only cultured with DMEM medium.The survival rate and apoptotic rate were detected respectively with MTT and TUNEL in NIH-3T3 cell sus-pension samples.The expression of cyclin D1,PTEN,NF-?B,HSP-70,BCl-2,BAX and caspase-3 were examined with immunocytochemistry and immunoblotting.Besides,20 Wistar rats were divided into control group and experimental group,the latter was given with quercetin in the doze of 0.13 mmol/kg.The levels of T-AOC,SOD,GSH-Px,GSH,MDA,NOS and NO2-/NO3-were detected both in the cleaved NIH-3T3 cells and in the plasma from both experimental and control animals prior to and post-1 h,2 h and after 24 h.Results When the Qb group was compared with H2O2 or Qa group,the survival rate was higher and the apoptotic rate was lower.When the H2O2 group was compared with C group,the expression of cyclin D1、PTEN or BCl-2 was down-regulated;while that of BAX、HSP-70、NF-?B or caspase-3 was up-regulated;the level of T-AOC,SOD,GSH-Px or GSH was decreased;that of NOS、NO2-/NO3-or MDA enhanced in the cleft NIH-3T3 cells.When the plasma level of the anti-oxidative enzyme system prior to-compared with post-1h and 2h-treatment with Q,the level of T-AOC,SOD,GSH-Px and GSH,especially the former two,were higher;MDA,lower;NOS or NO2-/NO3-promoted.However,the above parameters basically became normal 24 h after treatment with Q.Conclusion Quercetin down-regulates the promoted expression of HSP70,NOS,NO2-/NO3-and NF-?B etc.in H2O2-treatment NIH-3T3 cells.Qb could reverse the H2O2 damage effects more markedly.Moreover,the quercetin exerts anti-oxidant protective effect through modulating the anti-oxidative enzyme system both in vivo and in vitro.However,based on the cell heterogeneity in none-or pre/post-H2O2-treatment state,a difference in quercetin antioxidant response is noted.
