A method was developed for purification of 20S proteasome (20S core particle, CP) by combining differential centrifugations with nondenaturing polyacrylamide gel electrophoresis (native-PAGE), irrespective of species origins of CPs. CP purified from human erythrocytes was subjected to proteomic analysis by two-dimensional gel electrophoresis (2-DE) and matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS), revealing 33 spots of subunit isoforms with different molecular weights and isoelectric points, more than 14 constituent subunits. Furthermore, other four CPs were purified from yeast, mouse liver, two pancreatic cancer cell lines SW1990 and PANC-1 using this method mentioned above, and subjected to proteasome heterogeneity analysis by native/SDS-PAGE (native/sodium dodecyl sulphate polyacrylamide gel electrophoresis), together with CP from erythrocytes. The method described acts as a rapid and effective tool for CP isolations, and the results obtained may be served as a footstone for the investigations of species-dependent proteasome heterogeneity.

AIM: To detect whether N-acetyl-seryl-aspartyl-lysyl-proline (Ac-SDKP) inhibits epithelial-mes-enchymal transition in A549 cells induced by TGF-β1 through suppressing the expression of heat shock protein 27 (HSP27) and zinc finger proteins Snail (including SNAI1and SNAI2) which ultimately inhibited the deposition of type I and type III collagens.METHODS:The colocalizations of HSP27 and SNAI1/SNAI2 respectively on A549 alveolar epi-thelial cells induced by TGF-β1 were measured by confocal microscopy .The expression of HSP27, SNAI1 and SNAI2 at mRNA level was detected by real-time PCR.Western blotting analysis was used to detect the expression of HSP 27, SNAI1 and SNAI2 on epithelial-mesenchymal transition in A549 cells induced by TGF-β1 and also the deposition of type I and type III collagens in A549 cells transfected with HSP27shRNA prior to TGF-β1 stimulation.RESULTS: Compared with control group, TGF-β1 increased the expression of HSP27, SNAI1, SNAI2, type I and type III collagen, which decreased significantly followed by Ac-SDKP intervention.The expression of SNAI1, type I and type III collagen decreased signifi-cantly after transfected with HSP27shRNA in A549 cells, which had the similar effect on Ac-SDKP intervention.CON-CLUSION:Ac-SDKP inhibits the transition of cultured A 549 cells to myofibroblasts and attenuates collagen synthesis by suppressing the expression of HSP 27 and zinc finger proteins SNAI 1 and SNAI2.

Objective To observe the analgesic and anti-inflammatory effects of the water extract of Glycosmis citrifolia (Willd.) Lindl.on mice and explore the mechanism. Methods The analgesic and anti-inflammatory effects were evaluated by 0.7% acetic acid-induced writhing test,the hot plate test,tests of dimethylbenzene-induced ear swelling,1% carrageenan-induced paw edema,determination of PGE2 in inflammatory feet,0.6% acetic acid-induced increase in peritoneal capillary permeability and cotton ball granuloma. Results The water extract of Glycosmis citrifolia (Willd.) Lindl.at low,medium and high doses can reduce the acetic acid-induced writhing times (P<0.01 or P<0.05),increase the pain threshold of mice (P<0.01 or P<0.05), inhibit dimethylbenzene-induced ear swelling (P<0.01 or P<0.05),1% carrageenan-induced paw edema (P<0.01 or P<0.05) and PGE2 production (P<0.01),0.6% acetic acid-induced increase of peritoneal capillary permeability (P<0.05),and the de-velopment of cotton ball granuloma (P<0.01 or P<0.05). Conclusion The water extract of Glycosmis citrifolia (Willd.) Lindl.shows analgesic and anti-inflammatory effects on mice.

Objective:To study preoperative MRI imaging and its enhanced mode on tumor features in predicting microvascular invasion (MVI) in patients with hepatocellular carcinoma (HCC).Methods:The clinical data of patients with a solitary HCC who underwent MRI examination followed by surgical resection at the General Hospital of Ningxia Medical University from January 2017 to June 2019 were studied. The patients were divided into the MVI (+ ) and MVI (-) groups according to the findings on postoperative pathological diagnosis. The relationship between the rates of MVI and MRI tumor features including diffusion weighted imaging (DWI) signal, enhancement mode, enhancement type and other imaging characteristics were analysed.Results:Of 84 patients with HCC enrolled into this study, there were 65 males and 19 females. Their age (Mean±SD) was (54.94±11.51) years. MVI (+ ) was found in 46 patients and MVI (-) in 38 patients. The maximum tumor diameters (Mean±SD) of the two groups were (7.08±3.45) cm and (4.28±2.47) cm ( P<0.01). Single-factor analysis and comparison of imaging characteristics of the two groups of patients showed tumor DWI signal, tumor encapsulation, enhancement mode, tumor edge smoothness, abnormal enhancement around tumors, and intratumoral arteries were significantly different ( P<0.05); There were no significant differences in T 1WI signals, T 2WI signals, tumor periphery, and enhancement types between groups. After inputting MVI(+ ) as a risk factor into the logistic regression model, tumor maximum diameters >6.33 cm, type 3/4 enhancement mode, and unsmoothness of tumor edge were independent risk factors (all P<0.05). Through combined diagnosis using ROC curve analysis with a cut-off value of 0.53, the area under the curve was 0.881, the sensitivity 0.870, specificity 0.789, and the Youden index 0.659. Conclusion:The multivariate logistic regression model and combined diagnosis using ROC curve analysis improved the diagnostic efficacy of MVI in its prediction of HCC on imaging studies. The risk predictors were easy to use and to promote in clinical practice.

Ongoing study on the chemical constituents of the roots of Macleaya microcarpa led to the isolation of eight compounds of derivatives of triterpenes and organic acids in addition to some previously identified benzophenanthridines. The eight compounds were identified by spectroscopic methods as well as comparison with literature values as 1-oxo-2, 22 (30)-hopandien-29-oic acid (1), 3-oxo-12-oleanen-30-oic acid (2), 3α-hydroxy-12-oleanen-30-oic acid (3), 3β-hydroxy-12-oleanen-30-oic acid (4), ferulic acid (5), ferulic acid 4-O-β-D-glucoside (6), 3-O-feruloylquinic acid (7), and methyl 3-O-feruloylquinate (8). Of which, 1 is a new triterpenoid of hopanes and 2-8 are isolated from M microcarpa for the first time. In order to discover natural active compounds as potential agents of anti-ulcerative colitis (UC), an in vitro drug high-throughput screening model targeted x-box-binding protein 1 (xbp1) was employed to evaluate the activity of the major chemical constituents of M microcarpa. The result confirmed that two dihydrobenzophenanthridines, dihydrosanguinarine (9) and dihydrochelerythrine (10), showed a certain activity on activating the transcription of xbpl, a transcription factor (TF) associated with the occurrence, development, and potential treatment of UC, with their relative activating ratios being 1.76 and 1.77 times, respectively, as compared with control group.

[Abstract ] Objective Silicosis is one of the most serious occupational diseases in China .In this study,we explored the reg -ulatory effect of N-acetyl-seryl-aspartyl-lysyl-proline ( Ac-SDKP ) on angiotensin ( Ang ) Ⅱ-induced extracellular signal-regulated ki-nase ( ERK1/2) and Jun N-terminal kinase ( JNK) signals and its inhibitory effect on the differentiation of human embryonic lung MRC-5 fibroblasts to myofibroblasts via Ang Ⅱ-induced ERK1/2 and JNK signals . Methods Human embryonic lung MRC-5 fibro-blasts were induced by Ang Ⅱand pre-treated with the JNK signal inhibitor ( SP600125 ) , the ERK1/2 signal inhibitor ( PD98059 ) or Ac-SDKP.The proliferation of the cells was measured by MTT assay .The expressions of αS-MA, SRF, p-ERK1/2 and p-JNK were determined by immunocytochemical staining , and the expression levels of these proteins and collagen Ⅰwere detected by Western blot .Results The A value of Ang Ⅱ group (0.56 ±0.08) measured by MMT assay was 2.07 fold as control group ( 0.27 ±0.05 ). Pretreatment with SP600125 , PD98059 and Ac-SDKP, the A value were (0.39 ±0.02), (0.40 ±0.03) and (0.36 ±0 0.5) that had a statistical significance with Ang Ⅱgroup.The up-regulation of colla-gen type Ⅰ,α-SMA, SRF were induced by Ang Ⅱ by 4.50, 3.50 and 3.00 fold compared with control group.Moreover, the expression of p-ERK1/2 and p-JNK were increased as 6.71 and 7.90 fold as control. Pre-treatment with Ac-SDKP could inhibit p-JNK and p-ERK1/2 to 29.79% and 46.84% compared with AngⅡ group. Conclusion Ac -SDKP can inhibit the differentiation of human embryonic lung MRC-5 fibroblasts to myofibroblasts by regulating AngⅡ-induced JNK and ERK1/2 signals.

OBJECTIVETo explore the inhibition effect of N-acetyl-seryl-aspartyl-lysyl-proline (Ac-SDKP) on myofibroblast differentiation of MRC-5 human fetal lung fibroblasts induced by angiotensin (Ang) II.

METHODSThe study was divided into 2 step: (1) MRC-5 human fetal lung fibroblasts was induced for 48 h at different dose of Ang II and at different time point by 100 nmol/L Ang II. Then the expression of collagen type I and α-smooth muscle actin (α-SMA) were mesaured by western blot. (2) MRC-5 human fetal lung fibroblasts were divided into 4 group: (1) control, (2) Ang II, (3) Ang II+Ac-SDKP, (4) Ang II+8-Me-cAMP (a specific activator of Epac). The α-SMA expression was observed by immnocytochemical stain. The protein expression of collagen type I, α-SMA, serum response factor (SRF), myocardin-related transcription factor (MRTF)-A, exchange protein directly activated by cAMP (Epac) 1, 2 were measured by Westen blot.

RESULTSMyofibroblast differentiation could be induced by Ang II from MRC-5 cells with a dose- and time-dependent manner. The up-regulation of SRF and MRTF-A were observed in MRC-5 cells induced by Ang II and accompanied with collagen I and α-SMA increased. Pre-treatment with 8-Me-cAMP or Ac-SDKP could attenuated all this changes induced by Ang II, and promoted the expression of Epac1.

CONCLUSIONAc-SDKP can inhibit the myofibroblast differentiation of MRC-5 cells induced by Ang II via Epac1 activating.

Actins ; Angiotensin II ; Cell Differentiation ; drug effects ; Collagen ; Collagen Type I ; Cyclic AMP ; analogs & derivatives ; Fetus ; cytology ; Fibroblasts ; cytology ; Guanine Nucleotide Exchange Factors ; Humans ; Lung ; cytology ; Myofibroblasts ; drug effects ; Oligopeptides ; pharmacology ; Serum Response Factor ; Trans-Activators

OBJECTIVETo perform a comparative proteomic analysis for identification of pulmonary proteins related to the progression of silicosis and anti-fibrotic effect of N-acetyl-seryl-aspartyl-lysyl-proline (Ac-SDKP).

METHODSBronchial instillation of SiO₂powder (for 4 or 8 weeks) was applied in rats to establish a silicosis model. Ac-SDKP treatment was performed before (prevention group) or after (treatment group) SiO₂instillation. The control group was treated by bronchial instillation of sodium chloride solution of the same volume as SiO₂powder for 4 or 8 weeks. Proteins in lung tissue were separated by two-dimensional gel electrophoresis and stained with colloidal Coomassie brilliant blue. The gel images were scanned with the Lab Scan III system and analyzed with Imagemaster 6.0. The protein spots with significant differences between two groups (i.e., P value was less than 0.05 in One-way ANOVA) and with a change in volume over 30% were defined as differential proteins. Comparison was performed between the silicosis group and control group after 4 or 8 weeks, between the Ac-SDKP treatment group and silicosis group after 8 weeks, and between the Ac-SDKP prevention group and silicosis group after 8 weeks. The differentially expressed proteins were subjected to in-gel digestion with trypsin and MALDI-TOF-MS and Mascot search engine analysis to identify these proteins.

RESULTSThirty-three differential proteins were identified. In comparison with the control group (4 weeks), the silicosis group (4 weeks) had 17 up-regulated proteins and 11 down-regulated proteins. In comparison with the control group (8 weeks), the silicosis group (8 weeks) had 16 up-regulated proteins and 12 down-regulated proteins. In comparison with the silicosis group (8 weeks), the Ac-SDKP treatment group had 5 up-regulated proteins and 6 down-regulated proteins, and the Ac-SDKP prevention group had 8 up-regulated proteins and 10 down-regulated proteins.

CONCLUSIONCritical regulatory proteins related to silicotic fibrosis and anti-silicotic effect of Ac-SDKP have been identified. These proteins may play an important role in proliferation, apoptosis, inflammation, epithelial-mesenchymal transition, and signal transduction in silicosis.

Animals ; Disease Models, Animal ; Lung ; metabolism ; Male ; Oligopeptides ; therapeutic use ; Proteome ; metabolism ; Rats ; Rats, Wistar ; Silicosis ; drug therapy ; metabolism

This paper was aimed to study the safety of moxibustion with FMEA method.Failure mode and effects analysis (FMEA) were used in every aspect of the operation process of moxibustion.And the local skin temperature was measured in 80 patients treated with moxa box moxibustion.The results showed that the skin temperature reached the highest when the moxibustion was given for 15 min,which was in consistence with the patients' chief complaints and their tolerances.It indicated that moxibustion for 15 min was the best moxibustion amount.Meanwhile,inspection should be made to avoid burning.After the application of FMEA,the RPN of the inspection activities,the temperature and distance of moxibustion were significantly decreased (P ＜ 0.05).It was concluded that the application of FMEA management mode strengthened the risk management of moxibustion treatment,standardized treatment process,provided the basis for the temperature and distance of moxibustion,and ensured the safety and efficacy of treatment.

Objective:To explore the value of clinical data and MRI image features in predicting and analyzing the degree of differentiation of hepatocellular carcinoma (HCC).Methods:The clinical and imaging data of 180 patients with surgical outcomes of HCC from March 2015 to June 2019 in the General Hospital of Ningxia Medical University were retrospectively analyzed. Alpha-fetoprotein (AFP)、aspartate aminotransferase (AST)、D-dimer、clinical stage、tumor length、apparent diffusion coefficient(ADC)、enhancement types and so on the clinical and imaging data of the poorly differentiated and non-differentiated HCC were compared and analyzed. Multivariate logistic regression was used to predict independent risk factors for poorly differentiated HCC.Results:Of the 180 HCC patients, 121 were moderately and highly differentiated, and 59 were poorly differentiated. Univariate analysis showed that the patient’s age, gender, AFP, AST, D-dimer level, clinical stage, Child-Pugh score, tumor length, whether the capsule was complete, tumor apparent diffusion coefficient, the maximum level ADC value, enhancement type with HCC differentiation degree were correlated(all P<0.05). Multivariate logistic regression analysis showed that the patients' gender ( OR=4.524, P<0.05), clinical stage ( OR=5.598, P<0.05), D-dimer ( OR=8.576, P<0.05), HCC diameter ( OR=0.498, P<0.05), enhancement types ( OR=2.988, P<0.05), tumour ADC value ( OR=0.059, P<0.05) were independent of poorly differentiated HCC risk factor. Conclusion:MRI image features can be used as an effective indicator to predict the degree of HCC differentiation before surgery. It is more valuable to accurately predict the degree of HCC combined with D-dimer and AFP value.