Adolescent ; Adult ; Carbon Monoxide Poisoning ; epidemiology ; China ; epidemiology ; Female ; Humans ; Male ; Middle Aged ; Young Adult

Animals ; Male ; Particle Size ; Rats ; Rats, Sprague-Dawley ; Respiratory Distress Syndrome, Adult ; chemically induced ; Silicon Dioxide ; chemistry ; toxicity

Objective:To explore and analyze the points-selection rules in acupuncture treatment of mammary gland hyperplasia (MGH) by data mining and statistical method. Methods:Clinical literatures about the treatment of MGH with acupuncture published in the recent 16 years were retrieved from Chinese Journal Full-text Database (CJFD) and established into a database by Excel. The SPSS 20 version software and Clementine 12.0 version software were adopted to analyze the frequency and association rules of points-selection in the treatment of MGH with acupuncture. Results:The top 3 points used most frequently in acupuncture treatment of MGH were Danzhong (CV 17), Taichong (LR 3) and Zusanli (ST 36); points from the Stomach Meridian of Foot Yangming and Liver Meridian of Foot Jueyin were most commonly used; the commonly selected points were predominantly distributed in thoracic and abdominal regions and lower limbs; emphasis on the combination use of local and distal points; of the specific points, the five Shu-Transmitting points were mostly used; association analysis showed that the associations among Taichong (LR 3), Danzhong (CV 17) and Zusanli (ST 36) were the most significant. Conclusion: The data mining results substantially accord with the general rules of acupuncture-moxibustion theories in traditional Chinese medicine, able to reflect the points-selection principles and features in acupuncture treatment of MGH and provide evidence for the points selection in the treatment of MGH in acupuncture clinic.

Objective To investigate the expression of Foxp3~+ lymphocytes in breast carcinoma tissues and their correlation with other pathological factors,and to investigate the mechanism of action of Treg cells.Methods The expression of Foxp3~+ lymphocytes in the breast cancer tissue and non-cancerous tissue was detected by flow cytometry (FCM) in 30 breast carcinoma patients, and its correlation with other pathological factors was statistically analyzed by multiple linear regression analysis.The expression of TGF-β and IL-10 in the lymphocytes infiltrated in breast cancer tissue and non-cancerous tissue was measured by immunohistochemistry, and their correlation with the expression of Foxp3~+ lymphocytes was statistically analyzed by linear correlation dependability analysis. Results There was significant difference in the expression of Foxp3~+ lymphocytes between the malignant and non-cancerous breast tissues(P<0.05),and it was positively correlated with the clinical stage,blood vessel invasion and the matter of axillary lymph node metastasis(P<0.05). The expression of IL-10 in the tumor infiltrating lymphocytes was positively correlated with the expression of Foxp3~+ lymphocytes(P<0.05).Conclusion The expression level of Foxp3~+ lymphocytes is correlated with invasion and metastasis of breast carcinoma, and the IL-10 secreted by Foxp3~+ lymphocytes may be involved in this effect.Foxp3~+ lymphocytes can be used as an assistant marker for prediction and new therpeutic target of breast cancer.

Objective:To obtain the high expression of Herpes Simplex Virus Type 1(HSV-1)Glycoprotein D gene. Methods:The Herpes Simplex Virus Type 1(HSV-1)Glycoprotein D(gD1) gene fragment containing dominant antigen epitopes confirmed by computer analysis was cloned by PCR technical and inserted into plasmid vector pTrxA. Then the recombinant plasmid was transformed into Rosetta. The expressed product was analyze by SDS-PAGE. Results:930 bp gene fragment was amplified by PCR as anticipated. Nucleotide sequencing showed a 100 % homology with that of the published sequence in GenBank. The molecular weight of the expressed protein was about 48kDa, Western blotting indicated that the antigenicity of the protein was good. Conclusion:The plasmid pTrxA-gd1 was constructed and a high efficiency expression of the gd1 gene from Herpes Simplex Virus Type 1(HSV-1)strain was made. The expressed product shows a good antigenicity.

The deubiquitinating enzyme ubiquitin specific peptidase 15 (USP15) is regarded as a regulator of TGFβ signaling pathway. This process depends on Smad7, the inhibitory factor of the TGFβ signal, and type I TGFβ receptor (TβR-I), one of the receptors of TGFβ. The expression level of USP15 seems to play vital roles in the pathogenesis of many neoplasms, but so far there has been no report about USP15 in psoriasis. In this study, immunohistochemical staining of USP15, TβR-I and Smad7 was performed in 30 paraffin-embedded psoriasis specimens and 10 normal specimens to investigate the expression of USP15, TβR-I and Smad7 in psoriasis and to explore the relevance among them. And USP15 small interfering RNA (USP15 siRNA) was used to transfect Hacat cells to detect the mRNA expression of TβR-I and Smad7. Of 30 cases of psoriasis in active stage, 28, 24 and 26 cases were positive for USP15, TβR-I and Smad7 staining, respectively. The positive rates of USP15 and Smad7 were significantly higher in psoriasis specimens than in normal skin specimens (44.1%±26.0% vs. 6.1%±6.6%, 47.2%±27.1% vs. 6.6%±7.1%), and positive rate of TβR-I (20.3%±22.2%) in psoriasis was lower than that in normal skin specimens (46.7%±18.2%). There was a significant positive correlation between USP15 and Smad7 expression, and significant negative correlations between USP15 and TβR-expression, an I d between TβR- and Smad7 expression I in psoriasis. After transfection of USP15 siRNA in Hacat cells, the expression of TβR-mRNA was up I -regulated and that of Smad7 was down-regulated. It is concluded that USP15 may play a role in the pathogenesis of psoriasis through regulating the TβR-I/Smad7 pathway and there may be other cell signaling pathways interacting with USP15 to take part in the development of psoriasis.

OBJECTIVETo construct the acid-sensitive potassium hannel-3(TASK3) eukaryotic expression plasmid and to establish a stable SH-SY5Y cell line expressing enhanced green fluorescent protein (EGFP)-tagged TASK3.

METHODSTASK3 coding region was subcloned into pEGFP-N1 plasmid to construct a recombinant vector alled pEGFP-TASK3. The correct recombinant expressing plasmid was transfected with X-feet transfection reagent to SH-SY5Y cells. The cell line stably expressiing EGFP tagged-TASK3 gene was established by screening with antibiotic G418 and fluorescence microscope. The expression and localization of the EGFP tagged-TASK3 fusion protein was detected by Western blot and confocal microscope. Exposure of the SH-SY5Y cell line expressing stably TASK3-eGFP fusion proteins was exposed to different pH media (7.0, 6.7, 6.4, 6.1) for 24 h, the cell viability was assessed with cell counting Kit-8 (CCK-8).

RESULTSAll the results of identification by PCR, digestion with restriction endonuclease and sequencing indicated that the recombinant eukaryotic expression plasmid pEGFP-TASK3 was constructed correctly. The stable SH-SY5Y cell line expressing EGFP tagged-TASK3 fusion protein was successfully established. Exposure of the wild type SH-SY5Y cells and the stable SH-SY5Y-GFP tag-TASK3 cell line to different pH media (7.0, 6.7, 6.4, 6.1) for 24 h, the cell viability of two group cells significantly reduced with pH declining, and the difference was statistically significant (P < 0.05). Compared with wild type SH-SY5Y cells, the cell viability of stable SH-SYSY-GFP tag-TASK3 cell line increased significantly with the same pH media, and the difference was statistically significant (P < 0.05).

CONCLUSIONThe eukaryotic expression vector pEGFP-TASK3 is successfully constructed and the cell line stably expressing TASK3-eGFP fusion is established which is important for their fundamental research and potential applications.

Blotting, Western ; Cell Line ; Gene Expression ; Genetic Vectors ; Green Fluorescent Proteins ; genetics ; Humans ; Plasmids ; Polymerase Chain Reaction ; Potassium Channels, Tandem Pore Domain ; genetics ; Transfection

OBJECTIVETo evaluate the antiandrogenic effect of heterocyclic fungicide dimethachlon and its mechanism.

METHODSA combination of in vivo and in vitro assays was selected. Hershberger assay was used to determine the antiandrogenic potential of dimethachlon in vivo. Six-week-old castrated male SD rats were administrated once daily for 7 days with testosterone propionate (TP, 100 micro g/d, sc) plus gavage doses of dimethachlon (50, 100 or 200 mg x kg(-1) x d(-1)), or procymidone (150 or 300 mg x kg(-1) x d(-1), positive control), or iprodione (100 mg x kg(-1) x d(-1), positive control), or flutamide (50 mg x kg(-1) x d(-1), positive control). Transcriptional activation assay in vitro was employed to determine the mechanism of antiandrogenic activity of dimethachlon. Human hepatoma liver cells HepG2 were transiently cotransfected with human androgen receptor (AR) expression plasmid and AR-dependent luciferase report plasmid. Transfected cells were exposed to various concentrations of dimethachlon or flutamide with or without dihydrotestosterone to induce the expression of luciferase gene.

RESULTSIn Hershberger assay, dimethachlon, as well as other known antiandrogens, caused decrease in weight of androgen dependent organs or tissues. In 200 mg/kg group, the weight of seminal vesicle, ventral prostate, dorsolateral prostate, Cowper's gland, and levator ani plus bulbocavernosus muscles decreased by 57.8%, 44.8%, 43.9%, 30.1%, and 34.1% respectively, but did not decrease in the vehicle control group. The order of their antiandrogenic potencies was: flutamide > procymidone > dimethachlon > iprodione. In transcriptional activation assay, dimethachlon could inhibit dihydrotestosterone-dependent AR activity in transfected HepG2 cells in dose-effect relationship. The inhibiting potency of dimethachlon was about 1/100 of that of flutamide.

CONCLUSIONDimethachlon has antiandrogenic effect, and acts as an AR antagonist. Its antiandrogenic potency is lower than flutamide and procymidone, but higher than iprodione.

Aminoimidazole Carboxamide ; analogs & derivatives ; pharmacology ; toxicity ; Androgen Antagonists ; pharmacology ; toxicity ; Androgens ; blood ; metabolism ; Animals ; Body Weight ; drug effects ; Bridged Bicyclo Compounds ; pharmacology ; toxicity ; Cell Line, Tumor ; Chlorobenzenes ; pharmacology ; toxicity ; Dose-Response Relationship, Drug ; Flutamide ; pharmacology ; toxicity ; Fungicides, Industrial ; pharmacology ; toxicity ; Humans ; Hydantoins ; Luciferases ; genetics ; metabolism ; Male ; Pesticides ; pharmacology ; toxicity ; Plasmids ; genetics ; Rats ; Rats, Sprague-Dawley ; Receptors, Androgen ; drug effects ; genetics ; metabolism ; Succinimides ; pharmacology ; toxicity ; Transfection

Antineoplastic Agents ; chemistry ; Capillary Electrochromatography ; Chemistry, Pharmaceutical ; Drug Interactions ; Electrophoresis, Capillary ; methods ; trends ; Electrophoresis, Microchip ; Pharmaceutical Preparations ; metabolism ; Pharmacokinetics ; Platinum ; chemistry ; Stereoisomerism

OBJECTIVETo study whether zearalenone (ZEA), a fungal estrogen, can transcriptionally up-regulate the expression of cytochrome 450 3A4 (CYP3A4) transcription by activating human steroid hormone and xenobiotic receptor (SXR).

METHODTransient cotransfection reporter gene assays were performed with human SXR expression plasmid and a reporter plasmid containing the SXR in the CYP3A4 gene promoter in HepG(2) cells.

RESULTSThe transcriptional induction of CYP3A4 by ZEA with a dose, time-dependent manner. ZEA at the concentrations of 0.01, 0.10, 1.00 and 10.00 micromol/L, respectively, could induce CYP3A4 with (1.50 +/- 0.21), (1.66 +/- 0.27), (3.04 +/- 0.82) and (3.96 +/- 1.16) folds, as compared with 0.1% DMSO. Results from a time-dependent study show that 1.00 and 10.00 micromol/L of ZEA for 12 to 48 hours could enhance the transcription of CYP3A4 with (3.69 +/- 1.34) and (5.18 +/- 1.50) folds, and 10.00 micromol/L of ZEA for 48 hours could induce the CYP3A4 gene expression (5.18 +/- 1.50) folds, as compared with 0.1% DMSO by activating human SXR.

CONCLUSIONZEA could induce the expression of the CYP3A4 gene transcription through activating SXR, possibly by affecting the other substrates of the CYP3A4, especially affecting the metabolism of drugs in the body.

Carcinoma, Hepatocellular ; pathology ; Cytochrome P-450 CYP3A ; Cytochrome P-450 Enzyme System ; biosynthesis ; genetics ; Genes, Reporter ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Transcription, Genetic ; drug effects ; Tumor Cells, Cultured ; Zearalenone ; pharmacology