Objective To construct RNAi eukaryotic expressing vectors of human transcription factor glioma-associated oncogene homolog 1 (GLI1) with pGCsi-U6-GFP plasmid and to identify its activity in interfering GLI1.Methods Three GLI1siRNA targeting GLI1 were designed and synthesized according to the GLI1cDNA sequence in GeneBank,and then were cloned into pGCsi-U6-GFP to construct the recombinant plasmids,and transformed into E.coli DH5a,then it was amplified and plasmids were extracted,which were further confirmed by PCR reaction and DNA sequencing,pGCsi-U6-siRNA-C was negative as control wector.Then recombinant plasmids pGCs-U6-GLI1siRNA-1,pGCs-U6-GLI1 siRNA-2,pGCs-U6-GLI1siRNA-3 pGCsi-U6-siRNA-C and a eukaryotic over-expression vector pEGFP-N1-GLI1 were co-transfected into HEK293 cells by Lipofectamine 2000 respectively.The ceils were collected at 48 h after transfection.Semi-quantitative RTPCR and Western Blot were performed to detect the expression of GLI1 mRNA and protein to screen the optimal vector which had the best interfering effect.Results A 369 bp fragment was amplified from all three recombinant plasmids,(pGCs-U6-GLI1 siRNA-1,pGCs-U6-GLI1 siRNA-2,pGCs-U6-GLIlsiRNA-3),showing that synthesized shRNA oligonucleotide fragments were correctly inserted into three recombinant plasmids,which were further confirmed by sequencing.Expression levels of GLIlmRNA and protein in cells in pGCs-U6-GLI1 siRNA-1,pGCs-U6-GLI1 siRNA-2,pGCs-U6-GLI1 siRNA-3 were 0.290 ± 0.011,0.421 ± 0.018,0.373 ±0.018,and 0.318 ± 0.026,0.443 ± 0.021,0.381 ± 0.018,which were significantly lower than those in negative control group (0.834 ± 0.022,0.818 ± 0.024,P =0.000),the inhibitory rates were 65.8 ％,50.7％,55.7％,and 63.9％,48.3％,53.9％.The interfering efficacy of pGCs-U6-GLIlsiRNA-1 was the strongest among the three recombinant plasmids.Conclusions RNAi eukaryotic vectors pGCs-U6-GLIlsiRNA-1,pGCs-U6-GLI1 siRNA-2,pGCs-U6-GLI1 siRNA-3 are successfully constructed and the optimal vector is identified,and this can provide a solid experimental foundation for further functional study of GLI1 gene.

Objective To investigate the expression of GLI1 and PTCH1 mRNA in pancreatic cancer and study its clinical significance. Methods Real-time fluorescence quantitative PCR (RFQ-PCR) was used to detect the expression of GLI1 and PTCH1 mRNA in 35 samples of pancreatic cancer tissues and 27 samples of adjacent normal pancreatic tissues, and the correlation of GLI1 and PTCH1 mRNA expression with clinical parameters was investigated. Results The relative expression of GLI1 mRNA in pancreatic cancer tissues was 1.12 ～ 3. 65 ( median 1.19), the relative expression of TCH1 mRNA was 1.82 ～ 4.36 ( median 2.36 ). The relative expression of GLI1 mRNA in adjacent normal pancreatic tissues was 0.23 ～ 2.76 ( median 0.87 ), the relative expression of PTCH1 mRNA was 1.11 ～ 2. 17 (median 0.58). Both the expression of GLI1 and PTCH1 mRNA in pancreatic cancer tissues were significantly higher than those in normal pancreatic tissues (P<0.05), and a positive correlation was found between GLIl and PTCH1 mRNA expression levels (P <0.05 ). The expression of GLI1 mRNA was significantly correlated with the differentiation degree and lymph node metastasis of pancreatic cancer (P < 0. 05). Conclusions GLI1 and PTCH1 may be involved in pancreatic carcinogenesis, and GLI1 may be related to invasion and lymph node metastasis of pancreatic cancer.

Objective To evaluate the therapeutic effect of PLGA-PEG-PLGA-5-fluorouracil temperature-sensitive hydrogel interstitial chemotherapy for pancreatic cancer by using endoscopic ultrasonography.Methods PLGA-PEG-PLGA-5-fluorouracil temperature-sensitive hydrogel in vitro release experiments were performed in the following procedures:determination of lixivium drug concentration and calculation of its emission.Fifty nude mice with the pancreatic cancer cell line SW1990 were randomly divided into 5 groups,10 in each group.Group A was intratumorally injected with PLGA-PEG-PLGA-5-fluorouracil temperature-sensitive hydrogel at 4mg/kg ; group B with PLGA-PEG-PLGA-5-fluorouracil temperature-sensitive hydrogel at lmg/kg; Group C was intratumorally injected with 5-fluorouracil at 4 mg/kg ; Group D with PLGAPEG-PLGA matrix at 4mg/kg ;and group E was the control group.Tumor growth and audio-visual images of the nude mice tumor nodules were observed before administration,and 3,7,10,14 days after.Tumor growth curve was also drawn.Animals were sacrificed at 14 days Tumors were weighed to calculate the inhibitory rate and stained for pathological study.Results 1,3,5,8,10,and 14-day release of 5-fluorouracil temperature-sensitive hydrogel were 21.6％,33.8 ％,44.3％,63.6％,76.3％ and 91.8％.Tumor sizes of group A and group B were significantly different from those of other groups (P ＜ 0.05).Ultrasound endoscopic image characteristics were correlated with pathological findings.Conclusion PLGA-PEG-PLGA-5-fluorouracil temperature-sensitive hydrogel is able to release for 14 days in vitro,which constantly inhibits human pancreatic cancer cell line SW1990.Intratumoral injection of the agent can significantly inhibit the growth of pancreatic cancer of nude mice.Additionally,gelatinous preparations fixes better than liquid and is of clinical value.Therefore,monitoring temperature-sensitive 5-fluorouracil hydrogel interstitial chemotherapy for pancreatic cancer with endoscopic ultrasound is convenient and safe.

OBJECTIVETo detect the toxicity and teratogenicity of 2, 2-dinitroethene-1, 1-diamine (FOX-7) in rats.

METHODS125 adult SD rats were randomly divided into five groups, which are negative control (0 mg/kg) , positive control (280 mg/kg aspirin) , and three dose groups (5, 15, and 45 mg/kg) . They were administrated by gavage once a day from the 5th days to 19th days after pregnancy. The weight changes and toxicity of pregnant rats are recorded within the study, and the skeleton and internal organs malformations are detected by the recommended methods.

RESULTSAfter 5 or 6 days being poisoned, the pregnant rats appear significantly toxicity symptoms, such as exciting, irritability, and so on. The net weight raise in high dose group is less than the negative group, while the numbers of dead foetus in median and high dose groups are both more than that of negative group. Comparing with the negative group, the body weight and body lenghth of foetus rats in median and high dose groups, and the tail lenghth in high dose group are lower significantly. There are no external malformations in negative group and three dose groups. However, the foetus of high dose group appear significant skeleton and internal organs malformation prevalences that are significant more than negative group, including lateral cerebral ventricles enlarged, which accounts for 9.17%, occipital bone lost, which accounts for 2.59%.

CONCLUSIONFOX-7 can induced maternal reproductive toxicity, foetus toxicity and teratogenicity hazards to rats.

Animals ; Body Weight ; Female ; Nitro Compounds ; toxicity ; Pregnancy ; Rats ; Rats, Sprague-Dawley ; Teratogens ; toxicity ; Toxicity Tests

Objective:To prepare loratadine nanosuspension lyophilized powder (LTD-NLP) and investigate the dissolution and stability. Methods:LTD-NLP was prepared by an anti-solvent precipitation method; the particle size and morphology were characterized by a laser nanometer particle sizer and a transmission electron microscope (TEM). The optimal lyophilized protective agent was screened out; the dissolution and stability of LTD-NLP were determined by HPLC. Results:The average particle size of LTD-NLP was 175.6 nm with PDI of 0.200, and the zeta potential was -56.3 mV. The shape of LTD-NLP was spherical. The nanosuspension lyophilized with 10% sucrose presented optimal properties. The dissolution of LTD-NLP was greater than raw material, and LTD-NLP had good stability at 4℃. Conclusion:The anti-solvent precipitation method for LTD-NLP is simple,and the product is expected to become a new preparation of loratadine.

Objective: To construct the cell models of TNF-α rs1800620 (SNP1) and TNF-α rs4645843 (SNP2) stable transgen-ic lines and study the effects of SNPs on the content of TNF-α in LX-2 cell line. Methods: TNF-α SNP1/TNF-α SNP2 mutant plasmid was synthesized by wild-type TNF-α targeting and then transfected into LX-2 cell line after lentivirus packaging. RT-qPCR and Western blot were used to detect the mRNA and protein levels of TNF-α in each recombinant cell. Results: The gene sequencing showed that plasmids were successfully constructed. Photographs from an inverted microscope showed that the recombinant cells had stable expres-sion of GFP. The results of RT-qPCR and Western blot showed that compared with the wild group, SNP1 and SNP2 could significantly reduce the mRNA (P<0. 05) and protein (P<0. 05) levels of TNF-α. Conclusion: The recombinant cells stably expressing TNF-α/TNF-α SNP1/TNF-α SNP2 LX-2 are constructed. SNP1 and SNP2 can significantly reduce the mRNA and protein levels of TNF-α in LX-2 cell line.

OBJECTIVETo screen out serum differential proteins between vinyl chloride monomer (VCM)-exposed workers and healthy controls by proteomics and analyze the functions of differential proteins, and to provide a basis for elucidating the pathogenesis of diseases caused by VCM exposure and searching for the protein biomarkers.

METHODSFasting venous blood was collected from 125 VCM-exposed workers and 40 healthy controls according to accumulated exposure doses. Proteins were precipitated by acetone precipitation. These proteins were identified by 2D-nano LC-ESI-TOF/MS and quantified by isobaric tags for relative and absolute quantitation. The functions of differential proteins were analyzed by gene ontology.

RESULTSA total of 596 proteins were identified, including 194 quantified proteins. There were 21 differential proteins according to the screening criteria (19 upregulated proteins and 2 downregulated proteins), including complement, apolipoprotein, and glycoprotein. The functions of these differential proteins were binding, enzyme regulator activity, catalytic activity, and transporter activity, and they were involved in the biological processes including immune system process and response to stimulus.

CONCLUSIONThe complement, apolipoprotein, and glycoprotein identified in the proteomics may be related to liver injury caused by VCM exposure, and they could be used as candidate protein biomarkers of diseases caused by VCM exposure.

Biomarkers ; blood ; Blood Proteins ; analysis ; Humans ; Liver ; injuries ; Occupational Exposure ; Proteins ; metabolism ; Proteomics ; Vinyl Chloride ; toxicity

OBJECTIVETo investigate whether pregnancy-induced hypertension (PIH) may increase oxidative stress in women with PIH, and to explore the mechanisms by which PIH may increase oxidative stress and potential free radical damage.

METHODSSeventy women with PIH and seventy women with uncomplicated normotensive pregnancy (UNP) whose age, nutritional conditions, levels of hemoglobin and albumin were all matched, were enrolled in a randomized controlled trial. Their plasma concentrations of nitric oxide (NO), vitamin C (VC), vitamin E (VE), and beta-carotene (beta-CAR) as well as their erythrocyte malondialdehyde (MDA), and activities of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPX) were determined by spectrophotometry.

RESULTSCompared with average values of the above experimental parameters in the women with UNP, the average value of erythrocyte MDA in the women with PIH significantly increased (P<0.0001), while the average values of plasma NO, VC, VE, and beta-CAR as well as those of erythrocyte SOD, CAT, and GPX in the women with PIH significantly decreased (P<0.0005-0.0001). The findings from partial correlation analysis (controlling for age) for 70 women with PIH showed that with elevated systolic blood pressure (SBP) and diastolic blood pressure (DBP), MDA value gradually increased (P<0.001), and NO, VC, VE, beta-CAR, SOD, CAT, and GPX values gradually decreased (P<0.02-0.001). The findings from reliability analysis for NO, VC, VE, beta-CAR, SOD, CAT, GPX, and MDA values used to reflect increased oxidative stress and potential free radical damage in women with PIH showed that the reliability coefficients (alpha, 8 items) = 0.7062, P<0.0001, and the standardized item alpha = 0.9116, P<0.0001.

CONCLUSIONThe findings in the present research suggest that pregnancy-induced hypertension can increase oxidative stress and potential free radical damage in women with pregnancy-induced hypertension.

Adult ; Case-Control Studies ; Female ; Free Radicals ; metabolism ; Humans ; Hypertension ; metabolism ; Oxidative Stress ; Pregnancy ; Pregnancy Complications, Cardiovascular ; metabolism

Objective To observe the effect of precision nursing based on visualization technolo-gy in treating patients with pulmonary nodule surgery.Methods A total of 186 patients with surgery for pulmonary nodules were randomly divided into control group and observation group,with 93 cases in each group.The control group was given routine nursing,while the observation group was given vi-sualization technology assisted precision nursing on the basis of the control group.The psychological status[the Self-rating Anxiety Scale(SAS)score and the Self-rating Depression Scale(SDS)score]and postoperative recovery[score of the Quality of Recovery Scale with 15 items(QoR-15)]were compared between two groups,and the nursing satisfaction degree was compared between the two groups as well.Results After the intervention,the scores of SAS and SDS in both groups de-creased,and the score of each item in the observation group was significantly lower than that in the control group(P＜0.05);after intervention,the each item of the QoR-15 and the total score in the observation group were significantly higher than those in the control group(P＜0.05);the nursing satisfaction degree of the observation group was 97.8％,which was significantly higher than 89.2％in the control group(P＜0.05).Conclusion Precision nursing based on visualization technology can improve the adverse psychological state of patients with pulmonary nodule surgery,enhance the quality of postoperative recovery,and increase nursing satisfaction degree.

Objective To observe the effect of precision nursing based on visualization technolo-gy in treating patients with pulmonary nodule surgery.Methods A total of 186 patients with surgery for pulmonary nodules were randomly divided into control group and observation group,with 93 cases in each group.The control group was given routine nursing,while the observation group was given vi-sualization technology assisted precision nursing on the basis of the control group.The psychological status[the Self-rating Anxiety Scale(SAS)score and the Self-rating Depression Scale(SDS)score]and postoperative recovery[score of the Quality of Recovery Scale with 15 items(QoR-15)]were compared between two groups,and the nursing satisfaction degree was compared between the two groups as well.Results After the intervention,the scores of SAS and SDS in both groups de-creased,and the score of each item in the observation group was significantly lower than that in the control group(P＜0.05);after intervention,the each item of the QoR-15 and the total score in the observation group were significantly higher than those in the control group(P＜0.05);the nursing satisfaction degree of the observation group was 97.8％,which was significantly higher than 89.2％in the control group(P＜0.05).Conclusion Precision nursing based on visualization technology can improve the adverse psychological state of patients with pulmonary nodule surgery,enhance the quality of postoperative recovery,and increase nursing satisfaction degree.