OBJECTIVE: To establish a technical process for the separation and purification of total flavones from Licorice. METHODS: The static absorption capacity of macroreticular adsorptive resins D101、Hz-806、AB-83 for total flavones from licorice were compared. The macroreticular adsorptive resin columns on the Licorice extractives were eluted respectively with different concentrations of ethanol, then the content, the weight of residue and purity coefficient of flavones from Licorice in the eluant were detected. RESULTS: The optimal technological conditions were found in AB-8 as follows: flow rate=3ml/min, sample concentration=1.5 mg/ml, pH=5 and 80% ethanol was used as eluting agent. CONCLUSIONS: AB-8 macroreticular adsorptive resin can effectively separate and purify total flavones from licorice, the purity coefficient thus obtained being over 50%, which meets the requirement of the study of effective components of herbal medicine.

Objective To study the functional roles of 1,25(OH)2D3 in osteogenic differentiation of the dental papilla stem cells. Methods The dental papilla stem cells were isolated and cultured in medium supplemented with different con-centrations of 1,25(OH)2D3 (1, 10 and 100 nmol/L). MTT assay was used to detect the cell growth, and flow cytometry was used to detect the cell cycle. Western blot assay was used to detect protein expression levels of receptor activator of nuclear factor-κB ligand (RANKL), osteoprotegerin (OPG) and vitamin D receptor (VDR). After siRNA silencing VDR expression, protein levels of RANKL and OPG were detected. Results MTT and flow cytometry results showed that there were no sig-nificant differences in the cell proliferation between different concentrations of 1,25(OH)2D3 (1, 10 and 100 nmol/L) and con-trol groups (P>0.05). Western blot results showed that there were protein expressions of VDR, RANKL and OPG in control group. The protein expressions of VDR, RANKL and OPG were increased after adding 1,25(OH)2D3, in which the upward trend was the most significant in VDR. After VDR expression was silenced by siRNA, the protein expression levels of VDR, RANKL and OPG were decreased. Conclusion 1,25(OH) 2D3 affects the osteoblast differentiation process of the dental pa-pilla stem cells by adjusting the VDR expression.

Objective To study the effect of carbonated drinks on primary and permanent teeth replacement in Chil-dren. Method Dog tooth enamel samples were soaked in coca-cola, sprite and pure soda, and the calcium, phosphorus lev-el were analysed. Dental papilla stem cells were separated and cultured in the conditioned medium by adding three drinks. PCR and western blot were used to detect mRNA and protein levels of activator of nuclear factor-k B receptor ligand (RANKL), osteoprotegerin (OPG) and vitamin D receptor (VDR) , then the possible role of each gene and interactions rela-tionship were analyzed. Results Compared with saline, coca-cola and sprite showed their significantly decalcification and dephosphorization role, while plain soda water showed calcium and phosphorus protective effect. These three drinks had no effect on mRNA and protein levels of RANKL gene (P>0.05). Coca-cola and sprite can reduce OPG mRNA and protein lev-els, and at the same time increase transcription and expression of the VDR gene. Plain soda water has no effect on the OPG gene manifestation, but can significantly reduce the transcription and translation level of the VDR gene. Conclusion Car-bonated drinks may affect the dental health of the children's primary and permanent teeth replacement by regulating bone re-lated gene expression and vitamin D receptor family.

Objective To investigate the function of tripartite motif protein 22 (TRIM22) and the interaction with eukaryotic translation initiation factor-4E (eIF4E) in the differentiation of NB4 cells, one kind of acute promyelocytic leukemia cells, which elucidates the mechanism of TRIM22 targeting to regulate eIF4E.Methods The model of NB4 cells inducing differentiation was established in vitro.The expression changes of gene and protein of TRIM22 and eIF4E were detected by using quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting.In addition, the effect on cell function and protein expression level of eIF4E after adopting electroporation technology to depress or over-express TRIM22 was detected by CCK-8 and flow cytometry.Finally, the interaction of TRIM22 and eIF4E was verified by using co-immunoprecipitation (CO-IP).Results The mRNA relative expression level of TRIM22 was gradually increasing from 1.01±0.15 to 30.98±2.79 (F=280.700, P=0.000), and the protein relative expression level was gradually increasing from 0.22±0.03 to 0.51±0.05 (F=51.430, P=0.000) after the all-trans-retinoic acid (ATRA) induction for NB4 cell.However, the mRNA relative expression level of eIF4E was gradually decreasing from 1.01±0.09 to 0.47±0.06 (F=20.520, P=0.000), with the same trend, the protein relative expression level was gradually decreasing from 0.97±0.02 to 0.64±0.09 (F=14.700, P=0.001).The expression level of PE-CD11b in the TRIM22 over-expression group with ATRA detected by flow cytometry [(78.80±2.00)%] was higher than that in the transfection group of empty vetor with ATRA [(58.70±2.70)%] (t=9.535, P=0.000) and the cotransfection group with ATRA [(61.60±3.80)%] (t=8.187, P=0.000).Meanwhile, the protein level of eIF4E changed reversely after over-expressing the gene level of TRIM22 (t=4.985, P=0.007).The CO-IP experiment was used to verify the interaction of TRIM22 and eIF4E.ConclusionTRIM22 is able to promote the cell differentiation during the process of NB4 cells differentiation.Furthermore, eIF4E is a target of TRIM22 for binding with, which plays an important role in depressing the expression of eIF4E.

Objective:To develop early gastric cancer (EGC) detection system of magnifying blue laser imaging (ME-BLI) model and magnifying narrow-band imaging (ME-NBI) model based on deep convolutional neural network, to compare the performance differences of the two models and to explore the effects of training methods on the accuracy.Methods:The images of benign gastric lesions and EGC under ME-BLI and ME-NBI were respectively collected. A total of five data sets and three test sets were collected. Data set 1 included 2 024 noncancerous lesions and 452 EGC images under ME-BLI. Data set 2 included 2 024 noncancerous lesions and 452 EGC images under ME-NBI. Data set 3 was the combination of data set 1 and 2 (a total of 4 048 noncancerous lesions and 904 EGC images under ME-BLI and ME-NBI). Data set 4: on the basis of data set 2, another 62 noncancerous lesions and 2 305 EGC images under ME-NBI were added (2 086 noncancerous lesions and 2 757 EGC images under ME-NBI). Data set 5: on the basis of data set 3, another 62 noncancerous lesions and 2 305 EGC images under ME-NBI were added(4 110 noncancerous lesions and 3 209 EGC images under ME-NBI and ME-BLI). Test set A included 422 noncancerous lesions and 197 EGC images under ME-BLI. Test set B included 422 noncancerous lesions and 197 EGC images under ME-NBI. Test set C was the combination of test set A and B (844 noncancerous and 394 EGC images under ME-BLI and ME-NBI). Five models were constructed according to these five data sets respectively and their performance was evaluated in the three test sets. Per-lesion video was collected and used to compare the performance of deep convolutional neural network models under ME-BLI and ME-NBI for the detection of EGC in clinical environment, and compared with four senior endoscopy doctors. The primary endpoint was the diagnostic accuracy of EGG, sensitivity and specificity. Chi-square test was used for statistical analysis.Results:The performance of model 1 was the best in test set A with the accuracy, sensitivity and specificity of 76.90% (476/619), 63.96% (126/197) and 82.94% (350/422), respectively. The performance of model 2 was the best in test set B with the accuracy, sensitivity and specificity of 86.75% (537/619), 92.89% (183/197) and 83.89% (354/422), respectively. The performance of model 3 was the best in test set B with the accuracy, sensitivity and specificity of 86.91% (538/619), 84.26% (166/197) and 88.15% (372/422), respectively. The performance of model 4 was the best in test set B with the accuracy, sensitivity and specificity of 85.46% (529/619), 95.43% (188/197) and 80.81% (341/422), respectively. The performance of model 5 was the best in test set B, with the accuracy, sensitivity and specificity of 83.52% (517/619), 96.95% (191/197) and 77.25% (326/422), respectively. In terms of image recognition of EGC, the accuracy of models 2 to 5 was higher than that of model 1, and the differences were statistically significant ( χ2=147.90, 149.67, 134.20 and 115.30, all P<0.01). The sensitivity and specificity of models 2 and 3 were higher than those of model 1, the specificity of model 2 was lower than that of model 3, and the differences were statistically significant ( χ2=131.65, 64.15, 207.60, 262.03 and 96.73, all P < 0.01). The sensitivity of models 4 and 5 was higher than those of models 1 to 3, and the specificity of models 4 and 5 was lower than those of models 1 to 3, and the differences were statistically significant ( χ2=151.16, 165.49, 71.35, 112.47, 132.62, 153.14, 176.93, 74.62, 14.09, 15.47, 6.02 and 5.80, all P<0.05). The results of video test based on lesion showed that the average accuracy of doctors 1 to 4 was 68.16%. And the accuracy of models 1 to 5 was 69.47% (66/95), 69.47% (66/95), 70.53% (67/95), 76.84% (73/95) and 80.00% (76/95), respectively. There were no significant differences in the accuracy among models 1 to 5 and between models 1 to 5 and doctors 1 to 4 (all P>0.05). Conclusions:ME-BLI EGC recognition model based on deep learning has good accuracy, but the diagnostic effecacy is sligntly worse than that of ME-NBI model. The effects of EGC recognition model of ME-NBI combined with ME-BLI is better than that of a single model. A more sensitive ME-NBI model can be obtained by increasing the number of ME-NBI images, especially the images of EGG, but the specificity is worse.

course of Internal Medicine and Surgery Nursing. Methods From September 2014 to July 2016, the Grade 2013 nursing students at the Sino American cooperative nursing class in the Nursing College of Wenzhou Medical University were selected as the experimental group by convenient sampling method, with a total of 59 people. The Grade 2012 nursing students in the Sino American cooperative nursing class were selected as the control group, with a total of 60 people. The nursing students in the control group received traditional teaching methods and the nursing students in the experimental group received the self participating teaching method. We compared students' self-learning ability, theoretical knowledge and comprehensive practical ability between the two groups. Results The total score of self-learning ability and the scores of all dimensions of experimental group were higher than those in the control group after class, and the differences were statistically significant (P< 0.05). The scores of theoretical knowledge and comprehensive practice ability of the experimental group were higher than those of the control group, and the differences were statistically significant (P< 0.05). Conclusions Self participation teaching method is a teaching method that helps teachers play a leading role in giving play to students' main body status. It helps to promote the development of nursing students' independent learning ability, and improve their theoretical level and nursing practice ability.

In order to study the functions of the HSPs (Heat shock proteins) of Eimeria tenella, we cloned a novel gene (which designated EtHSP) coding HSP of Eimeria tenella by RT-PCR and RACE (Rapid-amplification of cDNA ends). The full-length cDNA sequence of EtHSP was 1802 bp, containing a 1455 bp ORF (Open reading frame) (GenBank Accession No. FJ911605) encoding a deduced protein of 484 amino acids. Real-time PCR revealed that the mRNA level of EtHSP was much higher in sporozoites of E. tenella than other developmental stages (unsporulated oocysts, sporulated oocysts and merozoites). We constructed the recombinant plasmids pET28a(+)-EtHSP, then transformed it into E. coli BL21(DE3) for expression. SDS-PAGE indicated that the fusion protein was expressed in included bodies, with peak expression 6 h after induction by IPTG Western blotting revealed that the protein was specifically recognized by polyclonal antibodies against E. tenella, showing that the fusion protein was native antigen.
Animals ; Chickens ; Eimeria tenella ; genetics ; metabolism ; Escherichia coli ; genetics ; metabolism ; Heat-Shock Proteins ; genetics ; immunology ; metabolism ; Inclusion Bodies ; metabolism ; Male ; Molecular Sequence Data ; Open Reading Frames ; genetics ; Rabbits ; Recombinant Fusion Proteins ; genetics ; immunology ; metabolism ; Sequence Analysis, Protein

Objective To investigate the safety and efficacy of endoscopic submucosal dissection ( ESD) for early stage colorectal cancer and precancerous lesions. Methods Clinical data of 108 patients who received ESD for early stage colorectal cancer and precancerous lesions from December 2016 to June 2017 in Renmin Hospital of Wuhan University were analyzed. The lesion characteristics, postoperative pathological features, intraoperative and postoperative complications and postoperative follow-up outcomes were analyzed. Results The 108 patients all underwent ESD successfully with median operation time of 45 min. The rate of intraoperative perforation and postoperative delayed bleeding was 2. 8％ ( 3/108) and 2. 8％ (3/108), respectively. No postoperative delayed perforation occurred. Postoperative pathology showed that there were 41 cases ( 38. 0％) of tubular adenoma, 4 ( 3. 7％) villous adenoma, 39 ( 36. 1％) villous tubular adenoma [ including 41 ( 38. 0％) low-grade intraepithelial neoplasia and 16 ( 14. 8％) high-grade intraepithelial neoplasia] , 19 ( 17. 6％) adenocarcinoma, and 5 ( 4. 6％) other types. Among the 19 cases of adenocarcinoma, there were 11 cases of well-differentiated, 5 median-differentiated and 3 low-differentiated. The complete resection rate was 100. 0％ and the en bloc resection rate was 92. 3％ ( 100/108) . The mean follow-up time was 8. 1 months, and no recurrence was found during this period. Conclusion ESD is safe and effective in the treatment of early stage colorectal lesions. It is important to improve preoperative assessment, strengthen surgical skills, analyze postoperative pathological features and regularly follow up to guarantee the treatment quality of ESD.

Objective:To establish a mice model of inflammatory bowel disease (IBD) induced by dextran sulfate sodium (DSS), and to analyze the changes in intestinal inflammation and macrophage subsets at different stages, so as to find a new target for the treatment of IBD.Methods:Thirty male C57BL/6 mice of 6-8 weeks were randomly divided into control group, activation stage group and resolution stage group.The latter 2 groups were given 25 g/L DSS for 5 consecutive days to establish the IBD model.After 5 days, the mice were given filtered and sterilized water and sacrificed on the 10 th and 15 th day, respectively.Colon inflammation in mice was evaluated, including body weight, disease activity index (DAI) score, changes in colon length, histopathology and histopathological score.Then the expression levels of interleukin (IL)-1β, IL-6, tumor necrosis factor (TNF)-α, transforming growth factor (TGF)-β in colon tissues were detected by quantitative real-time PCR(qPCR). Finally, the changes of intestinal macrophage subsets were detected by flow cytometry. Results:The colon inflammation of mice in the activation stage group was significantly more severe than that in the control group, while the colon inflammation of mice in the resolution stage group was reduced.The colon length of mice in the activation stage group was (5.94±0.40) cm, which was significantly shorter than that in the control group [(7.25±0.29) cm], and the situation was slightly improved in the resolution stage with the colon length of [(6.87±0.95) cm], and the differences were statistically significant (all P<0.05). The mRNA expression levels of pro-inflammatory cytokines IL-1β, IL-6 and TNF-α in the activation stage were 53.40±6.58, 117.69±30.78 and 2.52±0.25, respectively, which were significantly higher than those in the control group (1.00±0.13, 1.00±0.39, 1.00±0.10); the mRNA expression levels of IL-1β, IL-6 and TNF-α in the resolution stage were 2.51±0.13, 5.43±0.51 and 1.73±0.14, respectively, which were significantly lower than those in the activation stages(all P<0.05). The expression level of anti-inflammatory cytokine TGF-β in the resolution stage was 2.41±0.17, which was significantly higher than that in the activation stage (0.94±0.12), and the diffe-rence was statistically significant ( P<0.05). During the progression of IBD, there were 3 groups of macrophages in the lamina propria of intestinal mucosa of mice, of which the number of F4/80 lowCD 64-MHCⅡ - macrophage subset with the lowest maturity increased significantly in the activation stage of IBD, accounting for (10.68±4.62)%, and it decreased and returned to the normal level in the resolution stage, accounting for (4.63±1.06)%, and the difference was statistically significant ( P<0.05). Conclusions:Macrophages play an important role in the progression of IBD, the hindrance of maturation and development may be the main cause of inflammatory injury in the activation stage of IBD, and the transformation of macrophage subsets may become a new target for the treatment of IBD.

Objective:To compare the ability of deep convolutional neural network-crop (DCNN-C) and deep convolutional neural network-whole (DCNN-W), 2 artificial intelligence systems based on different training methods to dignose early gastric cancer (EGC) diagnosis under magnifying image-enhanced endoscopy (M-IEE).Methods:The images and video clips of EGC and non-cancerous lesions under M-IEE under narrow band imaging or blue laser imaging mode were retrospectively collected in the Endoscopy Center of Renmin Hospital of Wuhan University, for the training set and test set for DCNN-C and DCNN-W. The ability of DCNN-C and DCNN-W in EGC identity in image test set were compared. The ability of DCNN-C, DCNN-W and 3 senior endoscopists (average performance) in EGC identity in video test set were also compared. Paired Chi-squared test and Chi-squared test were used for statistical analysis. Inter-observer agreement was expressed as Cohen′s Kappa statistical coefficient (Kappa value).Results:In the image test set, the accuracy, sensitivity, specificity and positive predictive value of DCNN-C in EGC diagnosis were 94.97%(1 133/1 193), 97.12% (202/208), 94.52% (931/985), and 78.91%(202/256), respectively, which were higher than those of DCNN-W(86.84%, 1 036/1 193; 92.79%, 193/208; 85.58%, 843/985 and 57.61%, 193/335), and the differences were statistically significant ( χ2=4.82, 4.63, 61.04 and 29.69, P=0.028, =0.035, <0.001 and <0.001). In the video test set, the accuracy, specificity and positive predictive value of senior endoscopists in EGC diagnosis were 67.67%, 60.42%, and 53.37%, respectively, which were lower than those of DCNN-C (93.00%, 92.19% and 87.18%), and the differences were statistically significant ( χ2=20.83, 16.41 and 11.61, P<0.001, <0.001 and =0.001). The accuracy, specificity and positive predictive value of DCNN-C in EGC diagnosis were higher than those of DCNN-W (79.00%, 70.31% and 64.15%, respectively), and the differences were statistically significant ( χ2=7.04, 8.45 and 6.18, P=0.007, 0.003 and 0.013). There were no significant differences in accuracy, specificity and positive predictive value between senior endoscopists and DCNN-W in EGC diagnosis (all P>0.05). The sensitivity of senior endoscopists, DCNN-W and DCNN-C in EGC diagnosis were 80.56%, 94.44%, and 94.44%, respectively, and the differences were not statistically significant (all P>0.05). The results of the agreement analysis showed that the agreement between senior endoscopists and the gold standard was fair to moderate (Kappa=0.259, 0.532, 0.329), the agreement between DCNN-W and the gold standard was moderate (Kappa=0.587), and the agreement between DCNN-C and the gold standard was very high (Kappa=0.851). Conclusion:When the training set is the same, the ability of DCNN-C in EGC diagnosis is better than that of DCNN-W and senior endoscopists, and the diagnostic level of DCNN-W is equivalent to that of senior endoscopists.