Objective To evaluate the effect of intrathecal methotrexate on activation of spinal astrocytes in a rat model of bone cancer pain (BCP).Methods Sixty female unmated Sprague-Dawley rats,aged 5-7 weeks,weighing 150-180 g,were randomly divided into 3 groups (n =20 each) using a random number table:sham operation group (group S),BCP group (group P),and BCP + methotrexate group (group PM).BCP was induced by injecting Walker-256 cancer cells into the medullary cavity of tibia.On day 7 after BCP,methotrexate 100 μg (diluted to 15 μl in artificial cerebrospinal fluid) was injected intrathecally over 10 s in group PM,and artificial ccrebrospinal fluid 15 μ1 was injected intrathecally over 10 s in S and P groups.The mechanical paw withdrawal threshold (MWT) was measured at 1 day before BCP and 3,7,14 and 21 days after BCP.Five rats were sacrificed after measurement of the pain threshold at 7,14 and 21 days after BCP,and the lumbar enlargement segments of the spinal cord were harvested for detection of the expression of glial fibrillary acidic protein (GFAP) by immuno-histochemistry.Five rats were sacrificed after measurement of the pain threshold at 14 days after BCP,and the expression of GFAP in the spinal cord was detected by Western blot.Results Compared with group S,the MWT was significantly decreased at 3,7,14 and 21 days after BCP in C and CM groups,the expression of GFAP was significantly up-regulated at each time point after BCP in group C,and the expression of GFAP was significantly up-regulated at 7 and 14 days after BCP in group CM (P＜0.05).Compared with group C,the MWT was significantly increased,and the expression of GFAP was significantly down-regulated at 14 and 21 days after BCP in group CM (P＜0.05).Conclusion The mechanism by which intrathecal methotrexate reduces BCP may be related to inhibition of spinal astrocyte activation in the rats.

Objective To observe different effects of cyclic and continuous nutrition infusion on serum nutritional indicators, peripheral white blood cell counts (WBC), arterial partial pressure of oxygen (PaO2), arterial partial pressure of carbon dioxide (PaCO2) and acute physiology and chronic health evaluation Ⅲ (APACHE Ⅲ ) score in patients with mechanical ventilated respiratory failure (MVRF).Methods A cross-over and self-controlled trial was conducted in 48 patients with MVRF treated in a medical intensive care unit during December 2006 to June 2009, and continuous nutrition (group A) and cyclic nutrition (group B) were infused respectively for patients of the two groups.Serum levels of albumin (ALB), pre-albumin (PA), transferrin (TR), PaO2, PaCO2, WBC and APACHE Ⅲ score were measured for the patients with 24-hour continuous nutrition (group A) and 16-hour cyclic nutrition (group B)infusion.Effects of the two nutritional therapies were compared.Results After nutrition infusion, serum levels of ALB, PA and TR were (34±3)g/L, (196±28)mg/L and (2.1±0.3 ) g/L in group A, and (35 ±4) g/L, (198 ±25) mg/L and (2.0 ±0.4) g/L in group B, respectively; and PaO2 and PaCO2levels were (92 ± 12) mm Hg ( 1 mm Hg =0.133 kPa) and (42 ± 10) mm Hg in group A, and (91 ±9)mm Hg and (42 ± 10) mm Hg in group B, respectively.WBC and APACHE Ⅲ score were ( 11.8 ± 1.7) ×109/L and 38 ±7 in group A, and ( 12.6 ± 1.2) × 109/L and 40 ±6 in group B, respectively.Significant difference in serum levels of PA and TR was found between the two groups (PPA =0.019 and PTR =0.013),while there was no significant difference in other indictors between the two groups.Conclusions 24-hour continuous nutrition infusion for patients with MVRF can obviously improve their serum levels of PA and TR,but has no effect on serum level of ALB, PaO2, WBC and APACHE Ⅲ score in critical ill patients, as compared to those with 16-hour cyclic nutrition infusion.

Objective To identify the sounces of occupational stress and explore the relationship between occupational slreas and social support for nurses in ICU.Methods A descriptive and correlation design was used in this study.From July to October in 2007,159 nurses from 8 ICU and 160 nurses from common wards completed the questionnaires.Statistical analysis was performed for the investigation results.Results The ayes age score of occupational stress for nurses in ICU was (3.41±0.73),which was in a moderate to high level.The main stress was that they regarded nursing as a high risk career,maximums working intensity,always overload work and frequent night-shift.While the average score of occupational stress for nurses in common wards was (3.29±1.05).The main stress was that they regarded nursing as a high risk career,Score of occupational stress between nurses in ICU and com wards had no statistical difference(P＞0.05).There was a significant negative correlation between occupational stress and social support for nurses in ICU(r=-0.159,P＜0.05).And the sup port by friends was into the equalion by multi-linear regression.Conclusions The level of occupational stress for murses in ICU was fairly hish.Scientific management of human resource,reducing work pressure of nurses and workloed,exerting various social support will benefit for reducing the stress of nurses in ICU.

Objective: To study the impact of ticagrelor on inlfammatory factors in patients with unstable angina pectoris (UAP) after percutaneous coronary intervention (PCI).
Methods: Based on conventional medication, a total of 100 UAP patients after PCI were randomly divided into 2 groups for anti-platelet therapy:Treatment group, the patients received aspirin+ticagrelor and Control group, the patients received aspirin+clopidogrel. n=50 in each group, all patients were treated for 12 months. Plasma levels of inlfammatory factors were examined before treatment and 24h, 7d, 28d after the operation. The patients were followed-up for 12 months for ischemic events, bleeding events and adverse drug reactions.
Results: Before treatment, plasma levels of IL-6, IL-18, TNF-α, PDGF were similar between 2 groups (t=0.1356, 0.1668, 0.2473, 0.5780, all P>0.05). At 24h post-operation, the above inlfammatory factors were signiifcantly increased by peaks in both groups and decreased thereafter. At 24h, 7d and 28d post-operation, the levels of IL-6, IL-18, TNF-a, PDGF in Treatment group were all lower than Control group at each corresponding time point (at 24 post-operation, t=2.0856, 2.4399, 2.2217, 2.2053, all P<0.05), (at 7d post-operation, t=6.0978, 5.9705, 4.4631, 4.3963, all P<0.01) and (at 28d post-operation, t=9.3779, 9.7724, 5.5855, 6.1700, all P<0.01). The follow-up study presented that at 12 months post-operation, the ischemic events in Treatment group was lower than Control group (χ2=4.3956, P<0.05);there were no small hemorrhage or major bleeding events occurred;the slight adverse drug reaction was similar between 2 groups (χ2=0.1773, P>0.05).
Conclusion: Compared to clopidogrel, ticagrelor could better decrease plasma levels of inlfammatory factors, it may reduce ischemic events without elevate bleeding events.

Objective To evaluate the efficacy and safety of magnesium aluminum suspension spraying in treating gastric ulcer after endoscopic submocosal dissection (ESD). Methods We made a prospective analysis of patients with gastric ESD surgery performed between January 2016 and June 2019. They were divided into control group, spraying group, and spraying + oral group on the basis of treatment method. All patients were followed up for 8 weeks to evaluate endoscopic ulcer healing and recovery rate. At the same time, postoperative symptoms and complications such as bleeding were also recorded. SPSS 24.0 software was used for statistical analysis. Results A total of 330 eligible cases were randomly divided into control group (124 cases), spraying group (108 cases), and spraying + oral group (98 cases). The results showed that the postoperative pain incidence in spraying group and spraying + oral group was significantly better than that in the control group (35.48% vs. 17.59%, P<0.01; 35.48% vs. 20.41%, P=0.01). The 4-week ulcer recovery rate was better in spraying + oral group than in the control group. It should be noted that ulcer healing rate achieved 100% for the antrum of patients in spraying + oral group. At the same time, the incidence of postoperative bleeding decreased, and no significant adverse drug reactions were observed during the follow-up. Conclusion Almagate suspension spraying is safe and effective in the treatment of upper gastrointestinal ESD postoperative ulcer. In addition, it can also relieve postoperative pain.

Objective To investigate the expression of DNA methyltransferase 3b (DNMT3B) in hepatocellular carcinoma (HCC) and its effect and mechanism on the proliferation,invasion and migration of HCC cells.Methods The expression of DNMT3B gene was detected by qRT-PCR in 46 cases of HCC tissues and corresponding adjacent tissues;the results and clinical pathological parameters were analyzed.SiRNA targeting DNMT3B was transfected into MHCC97-H cells by RNA interference (RNAi) technique.The mRNA and protein expression levels of related genes were detected by qRT-PCR and Western blot.The cell proliferation was measured by MTT assay,and the invasion and migration abilities were measured by Transwell assay.Results In 46 HCC patients,the expression of DNMT3B (73.91％) was significantly higher in HCC than in adjacent normal tissue.The high expression of DNMT3B gene was associated with histological type and tumor size of HCC (all P＜0.05).Inhibition of DNMT3B gene expression decreased proliferation,invasion and migration of MHCC97-H cells.Interference with DNMT3B gene increased the expressions of tumor suppressor genes RASSFA1,APC and MTSS1 at mRNA and protein levels.Conclusion DNMT3B is associated with the progression of HCC.It may inhibit the proliferation,invasion and migration of HCC cells by regulating the methylation of downstream tumor suppressor gene.

ABSTRACT:Objective To observe the influence of saikosaponin-d (SSd)on the proliferation and the function of autophagy of human hepatocellular carcinoma (HCC)cell line SMMC-7721 to explore the possible mechanisms. Methods SMMC-7721 was cultured invitro and then treated with SSd of various concentrations (5.0,7.5,10.0, 12.5,15.0 and 17.5 mg/L)for 24,48 and 72 h.We used MTT to detect cell proliferation,selected the optimal concentration and time,and detected the expressions of BECN1 at mRNA and protein levels by PCR and Western blot.Results The inhibition rate of the proliferation of SMMC-7721 cell line increased with the increase of the concentration of SSd,and the highest inhibition rate (60%)appeared when the concentration reached 12.5 mg/L. The expression of BECN1 in the group with SSd was obviously higher than that in the control group (P<0.05). 3-MA decreased not only the expressions of BECN1 at mRNA and protein levels but also the expression of BECN1 when used in conjunction with SSd.Conclusion The inhibiting function of SSd on SMMC-7721 presents a dependency between drug concentration and function time,basically in line with the drug dose-effect relationship. SSd induces the occurrence of autophagic cell death through up-regulating the expression of BECN1 ,thus inhibiting the proliferation of SMMC-7 7 2 1 .

Objective To investigate whether epithelial-mesenchymal transition (EMT)is involved in the anti-invasion and anti-metastasis effects of curcumin on MDA-MB-2 3 1 breast cancer cell line and further analyze the underlying mechanisms.Methods MTT method was used to detect the anti-proliferative effect of curcumin on MDA-MB-2 3 1 cell line induced by lipopolysaccharide (LPS).The morphological changes were determined by optical and transmission electron microscopy,respectively.The expressions of Vimentin,E-cadherin,NF-κB and Snail were analyzed using RT-PCR and Western blotting.Results Curcumin could inhibit the occurrence of LPS-induced EMT of MDA-MB-2 3 1 breast cancer cell line, decrease the expression of LPS-induced EMT marker Vimentin and increase the expression of E-cadherin, resulting in the inhibition of in vitro cell motility and invasiveness.These actions were mediated by inactivating NF-κB-Snail signaling pathways.Conclusion Our data provide a new perspective of the anti-invasion mechanism of curcumin,indicating that the effect is in part due to its ability to inhibit the EMT process.

Objective:To explore the expression of polypyrimidine tract-binding protein 1 (PTBP1) in gastric cancer (GC) tissues and GC cell lines, and the role of PTBP1 in the proliferation and metastasis of GC cells.Methods:From January to June in 2019 at The First Affiliated Hospital of Xi′an Jiaotong University, the cancer tissues and corresponding para-cancer tissues of GC patients underwent surgical resection were collected. The Kaplan-Meier Plotter database was used to analyze the survival of GC patients. The expression of PTBP1 was down-regulated by transfecting small interfering RNA (siRNA) in human GC cell lines SGC7901 and AGS with relatively high expression of PTBP1. The cells were divided into blank control group, negative control group, and PTBP1 knockdown group. The expression of PTBP1 at mRNA and protein level were detected by real-time fluorescence quantification polymerase chain reaction (RT-qPCR) and Western blotting. At 24, 48, 72 and 96-hour after transfection, the effect of PTBP1 on the proliferation of GC cells was observed by 3-(4, 5 dimethylthiazol)-2, 5 diphenyltetrazolium bromide (MTT) experiment. The changes of invasion and migration of GC cells after down-regulation of PTBP1 were detected by transwell assay. The expression changes of epithelial-mesenchymal transition (EMT) markers E-cadherin, N-cadherin and vimentin after down-regulation of PTBP1 in GC cells were determined by Western blotting. Indenpendent samples t test, analysis of variance and rank sum test were used for statistical analysis. Results:The Kaplan-Meier Plotter prognostic analysis showed that the overall survival of GC patients with high PTBP1 expression was shorter than that of GC patients with low PTBP1 expression (9.2 months, 6.2 months to 17.2 months vs. 19.0 months, 14.5 months to 28.4 months), and the difference was statistically significant ( Z=5.31, P<0.05). The results of RT-qPCR showed that in GC cell lines SGC7901 and AGS, the expression of PTBP1 at mRNA level of PTBP1 knockdown group was lower than that of blank control group and negative control group (SGC7901: 0.78±0.11 vs.3.10±0.19 and 2.99±0.23; AGS: 0.80±0.09 vs. 3.55±0.24 and 3.50±0.18), and the differences were statistically significant ( tSGC7901=10.57 and 8.08, tAGS=10.91 and 13.42; all P<0.01). The results of Western blotting indicated that in GC cell lines SGC7901 and AGS, the expression of PTBP1 at protein level of PTBP1 knockdown group was lower than those of blank control group and negative control group (SGC7901: 0.38±0.04 vs. 1.42±0.05 and 1.35±0.09; AGS: 0.17±0.02 vs. 1.52±0.08 and 1.38±0.45), and the differences were statistically significant ( tSGC7901=15.94 and 10.57, tAGS=16.60 and 20.80; all P<0.01). The results of MTT showed that at 48, 72 and 96-hour after transfection the absorbance values of PTBP1 knockdown group decreased by 0.25±0.01, 0.38±0.02, and 0.84±0.04 as compared with those of negative control group, and the decrease was the most significant at 96-hour after transfection, and the differences were statistically significant ( t=10.21、14.32, both P<0.01). The results of transwell experiment demonstrated that the number of invasion and migration cells of PTBP1 knockdown group were both less than that of the blank control group and the negative control group (SGC7901: 42.00±5.91 vs. 116.40±10.23 and 114.40±10.43; 39.60±6.77 vs. 125.80±11.51 and 122.40±5.90; AGS: 40.20±7.25 vs. 115.60±14.63 and 117.40±9.12; 36.00±5.20 vs. 122.40±12.10 and 125.40±12.74), and the differences were statistically significant ( tSGC7901=14.07, 13.50, 14.43 and 20.62; tAGS=10.27, 14.75, 14.68 and 16.76; all P<0.01). The results of Western blotting showed that the expression of E-cadherin of PTBP1 knockdown group was higher than that of the blank control group and the negative control group (SGC7901: 1.42±0.05 vs. 0.53±0.05 and 0.57±0.03; AGS: 1.34±0.04 vs. 0.54±0.03 and 0.61±0.01), however the expression levels of N-cadherin and vimentin were both lower than those of the blank control group and the negative control group (SGC7901: 0.50±0.03 vs. 1.64±0.05 and 1.46±0.07; 0.32±0.07 vs. 1.42±0.07 and 1.33±0.07; AGS: 0.37±0.06 vs. 1.47±0.04 and 1.36±0.04; 0.41±0.04 vs. 1.53±0.06 and 1.37±0.04), and the differences were statistically significant ( tSGC7901=11.63, 13.19, 18.83, 11.68, 11.43 and 10.43; tAGS= 15.02, 16.23, 14.67, 12.97, 14.45 and 17.18; all P<0.01). Conclusions:The expression levels of PTBP1 increase in GC tissues and cells, which may be involved in regulating the proliferation, metastasis and EMT of GC cells.

Objective To investigate the expression of Notch1 protein and its clinicopathologieal characteristics in colorectal carcinomas. Methods The expression and location of Notch1 protein were determined using immunohistochemical S-P method in 30 colorectal carcinomas and 18 normal colorectal tissues. Results The average positive rate of Notch1 protein was 45.5 ± 0. 7% in colorectal carcinomas and 6. 8 ± 0. 9% in normal colorectal tissues. There was significant difference between colorectal carcinomas and normal colorectal tissues (P ＜ 0. 05). The expression of Notchl protein in different age, sex, tumor site, nodal involvement and Dukes stage had no significant difference. The expression of Notch1 protein in different histology grade had significantly statistical difference. Conclusion Notch1 expression was associated with the development of colorectal carcinoma. Notch1 was the potential oncagenic gene. Nothe1 would become the new target of the colorectal tumor therapy in the future.