Objective To evaluate the role of nuclear factor kappa B (NF-κB) in sevofluraneinduced release of inflammatory factors in mouse microglias.Methods Microglial cells obtained from newborn C57BL/6 mice (aged 2-3 days) were seeded in 24-well plates (density 1 × 105 cells/ml, 1 ml/well) , a total of 80 wells.The cells were randomly divided into 4 groups using a random number table (n =20 each) : control group (group C);sevoflurane group (group S);NF-κB selective inhibitor pyrrolidine dithiocarbamate (PDTC) group (group P);PDTC + sevoflurane group (group P +S).In S and P+S groups, 4.1％ sevoflurane was inhaled for 6 h.In P+S group, 10 μmol/L PDTC was added at 1 h before sevoflurane inhalation.At 6 h of incubation with sevoflurane, NF-κB activity and expression and levels of interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) were determined.Results Compared with group C, the NF-κB activity and levels of IL-6 and TNF-α were significantly increased in group S, and no significant change in the above parameters was found in the other two groups.Compared with group S, the NF-κB activity and levels of IL-6 and TNF-α were significantly decreased in group P+S.There was no significant difference in NF-κB expression between the four groups.Conclusion NF-κB mediates sevoflurane-induced release of inflammatory factors in mouse microglias.

Objective To determine the serum level of chernokine CCL27 in patients with psoriasis vulgaris,and to analyse its clinical relevance.Methods A total of 61 patients(40 in progressive stage and 21 in stable stage)with psoriasis vulgaris,with an average disease duration of 37.97±14.34 years,were included in this study.Appropriate thempy was given to these patients.Serum samples were collected from the patients before and after therapy,as well as from 45 healthy human controls.ELISA was applied to examine the serum concentration of CCL27.Clinical severity of psoriasis vulgaris was assessed by psoriasis area and severity index(PASI)score.Results Serum level of CCL27 was 670.02±262.15 ng/L in psoriatic patients,compared to 373.10±92.84 ng/L in the controls(t=8.18.P＜0.01).Increased serum level of CCL27 was observed in patients with progressive psoriasis vulgaris compared to those with stable psoriasis (799.94±214.54 ng/L vs 422.57±135.53 ng/L,t=8.39,P＜0.01).After 8 weeks of therapy,a significant decrease was noticed in the serum level of CCL27 in patients who experienced≥70%reduction in PASI score(t=9.95,P＜0.01).but not in those experiencing a PASI reduction of＜70%(t=1.84,P＞0.05).The serum level of CCL27 was positively correlated with PASI score(r=0.58,P＜0.01).Conclusions The serum level of CCL27 is significantly elevated in patients with psoriasis vulgaris,and it is correlated with the disease severity.

Carcinoma, Non-Small-Cell Lung ; genetics ; Exons ; genetics ; Genes, erbB-1 ; genetics ; Humans ; Lung Neoplasms ; pathology

OBJECTIVETo determine the expression of Skp2 in different prostate cancer (PCa) cell lines and tissues, and explore its influence on the androgen receptor (AR) signaling pathway and development of castration-resistant prostate cancer (CRPC).

METHODSThe expression levels of Skp2 and AR in different PCa cell lines were detected by Western blot. After knockdown of Skp2 in the C4-2 and 22RV1 cells transfected with shRNA, the expressions of AR and P27 were determined and the activity of ARR3-Luc measured by dual-luciferase reporter gene assay following treatment with dihydrotestosterone (DHT). The expressions of AR and Skp2 in human naïve PCa or CRPC specimens were detected by immunohistochemical staining followed by analysis of their differences and correlation.

RESULTSThe Skp2 protein expression level was significantly higher in the C4-2 or 22RV1 cells than in the LNCaP cells. DHT treatment increased the expression of Skp2 in the C4-2 cells, but knock-down of Skp2 significantly up-regulated the expression of the well-known downstream protein P27 and down-regulated that of AR. Consistently, DHT treatment increased the activity of ARR3-Luc, while knockdown of Skp2 remarkably decreased it in the C4-2 and 22RV1 cells (P < 0.05). In addition, significantly higher expressions of Skp2 and AR were observed in the CRPC than in the naïve specimens (P < 0.05), with a positive correlation between the two proteins (r = 0.658 1, P < 0.05).

CONCLUSIONSkp2 can enhance the expression and transcription activity of the AR protein in CRPC cells or tissues and is promising to be a critical molecular therapeutic target.

Androgens ; pharmacology ; Cell Line, Tumor ; Dihydrotestosterone ; pharmacology ; Disease Progression ; Gene Knockdown Techniques ; Humans ; Male ; Neoplasm Proteins ; genetics ; metabolism ; Prostatic Neoplasms, Castration-Resistant ; metabolism ; Receptors, Androgen ; genetics ; metabolism ; S-Phase Kinase-Associated Proteins ; physiology ; Transcriptional Activation ; Up-Regulation

Objective To investigate the effect and regulation mechanism of mimic ischemia/reperfusion (I/R) cul-ture of human umbilical vein endothelial cells (HUVECs) on levels of tumor necrosis factor(TNF)-αand intercellular adhe-sion molecule (ICAM)-1. Methods HUVECs were randomly divided into four groups:control group (normal media cell cul-ture+control siRNA transfection), mimic I/R+control siRNA transfection group (HUVECs were transfected with control siR-NA, for 48 h ,and then received mimic ischemic media culture for 30 min followed by normal media culture for 4 h), normal culture+p65 siRNA transfection group and mimic I/R+p65 siRNA transfection group. The expression levels of TNF-αand ICAM-1 mRNA and protein were determined by real-time PCR and enzyme linked immunosorbent assay (ELISA), respec-tively. Results The mRNA and supernatant protein levels of TNF-αand ICAM-1 were significantly increased in mimic I/R culture group (4.96±0.16 and 3.33±0.30)μg/L than those of other tree groups (P<0.05). The level of ICAM-1 mRNA was significantly higher in I/R+p65 siRNA transfection group (1.87±0.21)μg/L than that of control group (1.58±0.15) μg/L and normal culture+p65 siRNA transfection group [(1.69±0.21)μg/L, P<0.05]. The levels of TNF-αand ICAM-1proteins were (329.98 ± 12.18) μg/L and (654.74 ± 64.79) μg/L in mimic I/R+control siRNA transfection group, which were significantly higher than those of other three groups (P<0.05). The levels of TNF-αand ICAM-1 proteins were (129.65±22.42)μg/L and (185.76±11.27)μg/L in mimic I/R+p65 siRNA transfection group, which were significantly lower than those of contro group [(183.50±11.77)μg/L and (280.43±13.76)μg/L, P<0.05]. Conclusion The silencing of p65 through transfection of p65 siRNA in HUVECs inhibited mimic I/R-induced mRNA and protein expressions of TNF-αand ICAM-1.

Objective To amplify the recombinant adenovirus vector carrying rat angiotensin Ⅱ type 2 receptor (AT2R) gene using human embryonic kidney (HEK) 293A cell lines and to construct a pancreatic islet βcell model overexpressing AT2R by transfecting the adenovirus vector into rat insulinoma (INS-1) cell lines.Methods Recombinant adenovirus vector Ad-G-AT2R-EGFP and control vector Ad-CMV-EGFP were amplified with HEK 293A cells and the titer of the adenovirus was detected .After both adenovirus vectors were transfected into INS-1 cells,AT2R and angiotensin Ⅱtype 1 receptor(AT1R) gene expressions were tested using real-time PCR, Western blotting, immunofluorescence staining and confocal laser-scanning microscopy .Results The titer of amplified Ad-G-AT2R-EGFP and Ad-CMV-EGFP was re-spectively 9 ×109 pfu/ml and 8 ×109 pfu/ml.Transfection of Ad-G-AT2R-EGFP into INS-1 cells induced an increase in AT2R mRNA expression in a dose-dependent manner , and significantly increased AT2R mRNA and protein expression compared with Ad-CMV-EGFP-or mock-transfection.Conclusion The recombinant adenoviral vector carrying AT2R gene is successfully amplified and an INS-1 cell model overexpressing AT2R is constructed by transient transfection , which can contribute to further study of the role of AT2R in pancreatic islet βcells.

Background and purpose:Antracycline combined with paclitaxel is more widely applied in breast cancer as neoadjuvant chemotherapy.There are differences in applications of different paclita~els.In this research,the efficacy and toxicity of neoadjuvant chemotherapy with ET,ED regimen were compared for the patients with stageⅢbreast cancer.Methods:64 cases of stageⅢbreast cancer patients were divided in two groups.Before surgery,one group had received ET(EPI ivgtt 60 mg/m~2 d_1?21,PTX ivgtt 175 mg/m~2 d_2?21),the other group had received ED(EPI ivgtt 60 mg/m~2 d_2?21,DOC ivgtt 75 mg/m~2 d_2?21)neoadjuvant chemotherapy for three weeks. Curative effect and side effects were evaluated after 2-4 cycles.Results:Total effective rate was 87.5%.Effective rate in ED group was 92.9%,and effective rate in ET group was 83.5%.There was no significant difference(P=0.253).In pCR cases,8 cases in ED group achieved pathologically complete response compared to 3 cases in ET group(P=0.033). The number of patients in ED group(24 cases)hadⅣ-Ⅴgrade pathology evaluation after chemotherapy,it was higher than that in ET patients(21 cases).There was a significant difference(P=0.017).In both groups side effects including hair loss,nausea and vomiting,liver dysfunction were similar.Incidence rate of peripheral neurotoxicity in ET group was higher than that in ED group(P=0.002).Incidence rates of leukopenia,skin rash and phlebitis in ED group were higher than that in ET group.There was a significant difference between two groups in the leukopenia(P=0.034). Conclusions:For the patients with stageⅢbreast cancer patients,both two regimens could achieve better curative effect.ET and ED regimen have similar effect.But in ED regimen,the number ofpCR cases was obviously higher than in ET group.In both groups side effects were similar.There were significant differences in terms of leukopenia and peripheral neurotoxicity,but the side effects could be tolerated.

Objective To investigate the distribution of high-arsenic drinking water in Honghu city of Hubei province in order to provide a scientific basis for prevention and control of endemic arsenic disease.Methods Investigations were made in 22 townships(towns,districts),68 natural villages of the drainage areas of the Dongjing River,the Neijing River and the Yangtse River in 2006 and 2007,with the townships(towns,districts)around Shahu town in Xiantao city as the focal point.1000 water samples were drawn each year,which was 10% of all the wells in every natural village.Using sampling investigation,water arsenic Was determined by half-quantitative fast reagent kit.All samples of water with arsenic exceeding the standard(≥0.03 mg/L)were re-determined according to state standard.Surveys on the disease was carried out in the villages(brigades)where arsenic exceeded the standard.Results A total of 2000 samples were surveyed from 68 natural villages,of which there were 401 samples from 48 villages exceeding the standard in a rate of 20.05%(401/2000).The highest arsenic content Was 0.71 mg/L.The high arsenic water sources were distributed mainy in the drainage areas of the Dongjing River and the Neijing River,but no patients with endemic arsenic disease were found.Conclusions The high arsenic water sources are distributed mainly in the drainage areas of the Dongjing River and the Neijing River.It is suggested that the interrelated government departments should take precise measures to impmve the quality of drinking water and ensure safe water to the residents in high arsenic areas.

Objective To investigate the apoptosis induction effect of Cantharidin on pancreatic cancer cell line PANC1 and CFPAC-1 and possible mechanism. Methods PANC1 and CFPAC-1 was treated with Cantharidin. Cell growth was determined by MTT. Apoptosis was measured by flow cytometry. Caspase activity was measured by using enzyme chemical method. Apoptosis-related gene expressions were determined by using RT-PCR and Western blotting. Results Cantharidin significantly inhibited the growth of pancreatic cancer cells PANC1, CFPAC-1 and induced apoptosis in a dose-dependent manner. Seventy-two hours after 10 μmol/L Cantharidin treatment, the inhibitory rates of PANC1, CFPAC-1 were (52.95 ± 6.34)％ and (71.21 ±6.30)％. Twenty-four hours after treatment, the early and later period apoptotic cell of PANC1 was increased from 7.35％ to 24.89％, from 6.36％ to 17.73％. The early and later period apoptotic cell of CFPAC was increased from 6.39％ to 24.70％, from 9.21％ to 12.58％ (P＜0.01). Activity of caspase 8 and caspase 9 in PANC1 cells was (155.8 + 11.5)％ and (194.6 ± 14.7)％ when compared with that of control group. Activity of caspase 8 and caspase 9 in CFPAC- 1 was ( 182.5 ± 24.3 ) ％ and ( 215.8 ± 12.2) ％when compared with that of control group ( P ＜ 0. 01 ). The expression of pro-apoptotic genes, TNF-α,TRAILR1, TRAILR2, Bad, Bak and Bid was elevated, the expression of anti-apoptotic Bcl-2 gene was decreased. Conclusions Cantharidin can induce apoptosis in pancreatic cancer cell lines by activating caspase,up-regulating the expression of pro-apoptotic genes and down-regulating the expression of anti-apoptotic genes.

Objective To study on the effects of Huaweishu granule on cisplatin-induced Beagle dog vomiting models and its mechanism.Methods Cisplatin-induced Beagle dog vomiting models were adopted as research object.The changes of vomiting frequency,latency,serum motilin content in the medulla oblongata and ileum,5-HT and substance P content of these models after using Hua weishu Granule were observed.Results ① Latent period of vomiting of the model group was (60.8±37.1)min; while this period was (137.3± 53.4)min,(122.8 ± 50.7) min,(l16.8±44.6)min,in the Huawei-shu low,medium and high dose group respectively,all showing a statistic difference than the model group (P＜0.05).②Frequency of vomiting in the model group was (270.3±51.8),while this frequence was (111.5±45.0) and (149.5±26.8) in Huawei-shu low and medium dose group respectively; ③ serum motilin in the model group was (0.354±0.098)ng/ml,while serum motilin in Huawei-shu low,medium and high dose group was (0.230±0.074) ng/ml,(0.235± 0.071) ng/ml,and (0.245± 0.062)ng/ml respectively,all lower than the model group (P＜ 0.05).④ Medulla oblongata 5-HT in the model group was (2.028 ±0.198)ng/ml,while medulla oblongata 5-HT in Huawei-shu low,medium and high dose group was (1.620±0.329)ng/ml,(1.194±0.386)ng/ml,and (1.269 ± 0.251) ng/ml respectively,all were lower than the model group (P＜0.05); ileum 5-HT in the model group was (1.634± 0.221)ng/ml,while ileum 5-HT in Huawei-shu low,medium and high dose group was (1.108±0.291)ng/ml,(1.194±0.386)ng/ml,and (1.269 ± 0.251) ng/ml respectively,all were lower than the model group(P＜0.05) ;⑤ ileal substance P content of the model group was (0.356±0.063)ng/ml,while ildeal P content in Huawei-shu low dose group was (0.274±0.064)ng/ml,lower than the model group than(P＜0.05).The medullary substance P content was (0.432±0.021)ng/ml in the model group,while medullary P content in Huawei-shu low,medium and high dose group was (0.370±0.040) ng/ml,(0.385±0.029) ng/ml,and (0.386± 0.041)ng/ml respectively,all were lower than the model group(P＜0.05).Conclusion Huawei-shu granule can prevent cisplatin-induced Beagle vomiting.