Objective To study the differentially expressed proteins and their biological behavior in colorectal carcinoma tissues and the normal colonic mucosa by proteomics and molecular biology techniques. Methods The technique of fluorescence two dimension differential gel electrophoresis (2-D DIGE) was used to analyze the expression of differential proteins in normal colorectal mucosa and primary cancer foci. Liquid chromatography with electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) was used to identify the differential proteins. Transfection experiment of colorectal cancer cells was performed with the differential protein cDNA, and the changes in cytobiological behavior were observed. Results Significant differences of protein expression levels were found by two-dimension electrophoresis. Eight differential protein spots were analyzed and identified. Human carbonic anhydrase Ⅱ and protein disulfide isomerase were detected in normal colorectal mucosa, but not in primary cancer foci. Phosphoglycerate kinase 1, fumarate hydratase and aldolase A were expressed in primary cancer. After transfection with human carbonic anhydrase Ⅱ cDNA, the abilities of Lovo cells were obviously reduced in invasiveness, chemotaxy motor and drug resistance. Conclusions Differences on protein expression levels are found between normal colorectal mucosa and primary cancer foci by 2-DE DIGE. The pathogenesis of colorectal carcinoma is related to the reduced expressions of carbonic anhydrase II and protein disulfide isomerase and enhanced expression of aldolase A. The technique of differential proteomics is useful in reaching a indepth understanding of the pathogenesis mechanisms of human colorectal cancer.

Objective To investigate the protective effects and molecular mechanism of peroxisome proliferate-activated receptor α(PPARα) activation on acute myocardial damage induced by isoproterenol (Iso) in rats. Methods Thirty male Wistar rats, weighting 160～180 g, were randomly divided into control group, Iso group, fenafibrate(FF) group(each n=10) according to physique quantity. Acute myocardial injury caused by Iso abdomen cavity injection induced ischemia was established and the protective effects of peroxisome proliferate-activated receptor α activation were accessed by the level of ereatine kinase(CK), lactic dehydrogenase(LDH) in serum as well as the activities of myoperoxidase(MPO) in myocardium, and the protein expressions of PPABα in myocardium by Western blot. Results The level of serum CK in control group, lso group and FF group, was (62.41±9.47),(101.71±11.05),(75.64±11.73)kU/L, respectively(F= 34.34, P<0.01). Whereas the level of serum CK in Iso group and FF group was higher than that in control group(P<0.01 or<0.05), the level of serum CK in FF group was lower than that in Iso group(P<0.01). The levels of LDH in these three groups were (5912.20±204.44), (6365.78±137.10), (6089.76±169.60) U/L, respectively(F= 17.54, P<0.01). Compared with the control group, the levels of LDH in Iso and Fir groups were significantly increased(P<0.01 or<0.05). But the level of LDH in FIr group was decreased compared with that in Iso group(P<0.01). The activities of myocardial MPO in these three groups were (1.95±0.10),(3.89±0.17),(2.49±0.19)U/g, espectively(F=391.68,P< 0.01). The activities of myocardial MPO in Iso and FF groups were higher than that in the control group (all P< 0.01), while the activities of myocardial MPO in FIr group were lower than that in lso group(P<0.01). The protein expressions of PPARα in myocardium of these three groups were 251.57±10.95,191.97±10.74,215.08±9.61, respectively(F=82.69, P<0.01). Conclusion PPARα activation by its actor FF can exert protective effects on the acute myocardial ischemia injury induced by lso in rats through inhibiting the release of inflammatory cell factors.

Aim of the present study is to investigate activation effect of nobiletin on cystic fibrosis transmembrane conductance regulator (CFTR) chloride channel activity. CFTR-mediated iodide influx assay and patch-clamp tests were done on FRT cells stably co-transfected with human CFTR and EYFP/H148Q. Nobiletin potently activated CFTR chloride channel activity in a dose- and time-dependent manner. The CFTR blocker CFTR(inh)-172 could completely reverse the effect. Preliminary mechanism study indicated that nobiletin activated CFTR chloride channel through a direct binding way. In addition, ex vivo tests done on mice trachea showed that nobiletin time-dependently stimulated submucosal gland fluid secretion. Nobiletin may be a therapeutic lead compound in treating CFTR-related diseases including disseminated bronchiectasis.

OBJECTIVEThis study was performed to determine whether recombinant human angiopoietin-1 (rhAng-1) decreases the permeability of the blood-brain barrier (BBB) in focal cerebral ischemia and reperfusion rats, whether RhAng-1 opens the BBB by affecting tight junction associated proteins zonnula occludin-1 (ZO-1), occludin and adherens junction protein vascular endothelial (VE)-cadherin.

METHODSThe rats were divided into eight groups randomly( n = 10): (1) sham-operated group; (2) ischemia group; (3)-(5) ischemia/reperfusion (middle cerebral artery occlusion and reperfusion (MCAO/R) 12 h, 48 h, and 7 days) and 0.9% saline groups; (6)-(8) ischemia/reperfusion (MCAO/R 12 h, 48 h, and 7 days) and rhAng-1 groups. The Bee permeability was assessed by Evans blue extravasation. The messenger RNA and protein expressions of ZO-1, occludin, and VE-cadherin were determined by reverse transcription-polymerase chain reaction, Western blot, and immunohistochemistry assays.

RESULTSThe BBB permeability and the brain infarct volume were significantly decreased after rhAng-1 injection. The expressions of ZO-1, occludin, and VE-cadherin were increased after rhAng-1 injection.

CONCLUSIONrhAng-1 may decrease the permeability of BBB in MCAO/R rats by upregulation of ZO-1, occludin, and VE-cadherin.

Angiopoietin-1 ; pharmacology ; Animals ; Antigens, CD ; metabolism ; Blood-Brain Barrier ; cytology ; metabolism ; Brain Ischemia ; metabolism ; Cadherins ; metabolism ; Endothelium, Vascular ; cytology ; drug effects ; Humans ; Male ; Occludin ; metabolism ; Rats ; Rats, Wistar ; Recombinant Proteins ; pharmacology ; Reperfusion Injury ; metabolism ; Zonula Occludens-1 Protein ; metabolism

Objective To explore the changes of Sema3A and it′s receptor Npl in temporal lobe epilepsy(TLE)rat brain and the roles in epileptogenesis mechanism.Methods TLE model was established with male healthy SD rats,in which mossy fiber sprouting(MFS)was verified using Neo-Timm staining method.Sema3A mRNA,Npl mRNA and protein was respectively analyzed by immunohistochemistry and in situ hybridization in the entorhinal cortex(EC)or dentate gyrus(DG)at different time after LiCL-PILO induced TLE.Results There were Mossy fiber sprouting(7d:0.70?0.42,15d:1.50?0.52,30 d:2.20 ?0.41,60 d:2.50?0.51)in DG inner molecular layer(IML)of TLE rat compared with those of controls (P

In this study, a combination of metabolomics and network pharmacology was used to study the pharmacodynamic substances and mechanism of action of Yiyi Fuzi powder (YYFZ) on rheumatoid arthritis (RA) rats. The animal experiments were conducted in accordance with the requirements of the Experimental Animal Ethics Committee of Tianjin University of Traditional Chinese Medicine (approval number: TCM-LAEC2021241). The metabolomic analysis using UPLC-Q-TOF/MS technique identified 22 metabolites, including arachidonic acid, tryptophan, linoleic acid, phenylalanine, as significant biomarkers for the treatment of RA with YYFZ, and they were significantly regressed after YYFZ treatment. The analysis of YYFZ blood components also revealed that 11 blood components, including hypaconitine, benzoylhypaconitine, and deoxyaconitine, may be the components that exert direct pharmacological effects in YYFZ in vivo, and further network pharmacological analysis of blood components obtained that YYFZ may exhibit anti-inflammatory effects through acting on PI3K/Akt signaling pathway, estrogen signaling pathway, vascular endothelial growth factor (VEGF) signaling pathway. The results of this study provide implications for the clinical application of YYFZ.

OBJECTIVETo explore the effects of stromal interaction molecule 1 (STIM1) on the expression of apoptosis-related proteins in prostate cancer PC-3 cells.

METHODSWe transfected the lentivirus vector STIM1-pGCSIL-GFP carrying STIM shRNA into human hormone-independent prostate cancer PC-3 cells, and 3 days later observed the transfection efficiency by fluorescence microscopy. At 7 days after transfection, we determined the expression of STIM1 in the PC-3 cells by RT-PCR and Western blot and those of apoptosis-related proteins Bcl-2, Bax, survivin and activated Caspase-3 by Western blot.

RESULTSAt 3 days, inverted microscopy revealed a transfection efficiency of > 80%. At 7 days, the STIM1 expression was significantly inhibited at both mRNA and protein levels. The Bcl-2/Bax rate was remarkably decreased as compared with that of the control group (0. 31 vs 1.24 ) , and the survivin expression was markedly reduced, 0. 14 times that of the relative expression in the control. However, the Caspase-3 cleavage was significantly activated, 1.52 times that of the control (P <0.05).

CONCLUSIONSTIM1 can be regarded as an oncogene in prostate cancer PC-3 cells. Inhibition of its expression can induce PC-3 cell apoptosis by reducing the Bcl-2/Bax rate, decreasing the survivin expression, and activating the Caspase-3 pathway.

Apoptosis ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Humans ; Inhibitor of Apoptosis Proteins ; metabolism ; Male ; Membrane Proteins ; genetics ; Neoplasm Proteins ; genetics ; Prostatic Neoplasms ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; RNA, Small Interfering ; genetics ; Stromal Interaction Molecule 1 ; Transfection ; bcl-2-Associated X Protein ; metabolism

Adult ; Bone Cysts, Aneurysmal ; diagnosis ; surgery ; Bone Neoplasms ; diagnosis ; surgery ; Chondroblastoma ; diagnosis ; surgery ; Female ; Humans ; Male ; Talus ; surgery ; Young Adult

OBJECTIVETo compare the difference in the efficacy on thoracic facet joint disorder between the combined therapy of electroacupuncture and manual reduction and the simple manual reduction.

METHODSOne hundred and sixty patients were randomized into an electroacupuncture and manual manipulation group (group A) and a simple manual manipulation group (group B), 80 cases in each one. In the group A, Ashi points and three pairs of Jiaji (EX-B 2) bilateral to the painful sites were selected. The perpendicular puncture was used at Ashi points, the oblique puncture was used at Jiaji (EX-B 2) and connected with electric stimulation for 20 min, additionally, the corresponding manual reduction was adopted at the sites of facet joint disorder. In the group B, the simple manual reduction was applied to the affected sites. Acupuncture was given once every day, the manual reduction was applied once every 10 days. The treatment of 10 days made one session. The efficacy was analyzed statistically at the end of two sessions of treatment. Before and after treatment, McGill pain scale was adopted for the value statistical analysis. PRI score, VAS score and PPI score of patients were calculated before and after treatment and compared in the two groups. The efficacy was compared between the two groups.

RESULTSThe curative rate was 56.3% (45/80) in the group A, which was better than 18.8% (15/80) in the group B (P< 0.01). The total effective rate was 95.0% (76/80) in the group A, which was better than 76.3% (61/80) in the group B (P<0.01). The scores of PRI, VAS and PPI after treatment were all improved significantly in the two groups (all P<0.05), in which, the results in the group A were better than those in the group B (PRI: 4.00 +/- 0.97 vs 5.44 +/- 1.16, VAS: 3.29 +/- 0.72 vs 3.87 +/- 0.81, PPI: 1.07 +/- 0.74 vs 1.64 +/- 0.90, all P<0.05).

CONCLUSIONThe combined therapy of electroacupuncture and manual manipulation achieves the superior efficacy on thoracic facet joint disorder as compared with the simple manual manipulation. The combined therapy relieves the symptoms of thoracic facet joint disorder and reduces the severity of disorder.

Acupuncture Points ; Acupuncture Therapy ; Adult ; Aged ; Electroacupuncture ; Female ; Humans ; Male ; Middle Aged ; Thoracic Diseases ; therapy

Objective To investigate the effects of CO_2 pneumoperitoneum on the expression of focal adhesion kinase (FAK) of gastric cancer MKN-45 cells. Methods CO_2 pneumoperitoneum with different pressures was simulated in vitro, and the gastric cancer MKN-45 cells were divided into test and control groups. In the test group, gastric cancer MKN-45 cells were cultured in CO_2 pneumoperitoneum with different pressures [5, 10 or 15 mm Hg (1 mm Hg =0.133 kPa)] for 4 hours. The condition of the cells exposed to CO_2 pneumoperitoneum with a pressure of 15 mm Hg was observed at 0.5, 2 and 4 hours. Gastric cancer MKN-45 cells in control group were cultured at normal atmospheric pressure. The expression of FAK and phosphorylated FAK (FAK Tyr397) of each group was detected by Western blot. Multiple-group analysis was done by one-way ANOVA, and intergroup comparison was done by LSD test. Results In CO_2 pneumoperitoneum with pressures of 5, 10, 15 mm Hg, the expression of FAK was 2.14±0.17, 2.07±0.21 and 2.52±0.26, respectively, and the expression of FAK Tyr397 was 1.82±0.28, 1.93±0.52 and 3.71±0.37, respectively. The expression of FAK and FAK Tyr397 in the control group was 2.43±0.46 and 1.71±0.23, respectively. We found significant differences between the 2 groups (F = 2.171, 26.951, P < 0.01). After gastric cancer MKN-45 cells being treated for 0.5, 2 and 4 hours in CO_2 pneumoperitoneum with a pressure of 15 mm Hg, the expression of FAK Tyr397 was 3.41±0.44, 4.12±0.56 and 5.24±0.41 respectively, which is also significantly different (F =116.119, P < 0.01). The expression of FAK Tyr397 was back to 0.72±0.16 1 hour after the release of CO_2. Conclusions CO_2 pneumoperitoneum with different pressures can not promote the expression of FAK in gastric cancer MKN-45 cells which had been cultured for 4 hours, but can activate FAK through promoting its phosphorylation. The degree of FAK phosphorylation increases with pressure and time, and the activity of FAK decreases to pretreatment level rapidly once pressure is released.