Interpretation and Elaboration for the ARRIVE Guidelines 2.0—Animal Research: Reporting In Vivo Experiments (V) / 实验动物与比较医学

Improving the reproducibility of biomedical research results is a major challenge. Transparent and accurate reporting of the research process enables readers to evaluate the reliability of the research results and further explore the experiment by repeating it or building upon its findings. The ARRIVE 2.0 guidelines, released in 2019 by the UK National Centre for the Replacement, Refinement and Reduction of Animals in Research (NC3Rs), provide a checklist that is applicable to any in vivo animal research report. These guidelines aim to improve the standardization of experimental design, implementation, and reporting, as well as enhance the reliability, repeatability, and clinical translation of animal experimental results. The use of the ARRIVE 2.0 guidelines not only enriches the details of animal experimental research reports, ensuring that information on animal experimental results is fully evaluated and utilized, but also enables readers to understand the content expressed by the author accurately and clearly, promoting the transparency and completeness of the fundamental research review process. At present, the ARRIVE 2.0 guidelines have been widely adopted by international biomedical journals. This article is based on the best practices following the ARRIVE 2.0 guidelines in international journals, and it interprets, explains, and elaborates in Chinese the fifth part of the comprehensive version of the ARRIVE 2.0 guidelines published in PLoS Biology in 2020 (the original text can be found at https://arriveguidelines.org). This section includes the items 6-11 of Recommended 11 section, covering "Animal Care and Monitoring", "Interpretation/Scientific Implications", "Generalisability/Translation", "Protocol Registration", "Data Access" and "Declaration of Interests". Its aim is to promote a comprehensive understanding and use of the ARRIVE 2.0 guidelines among domestic researchers, to enhance the standardization of experimental animal research and reporting, and to promote high-quality development of experimental animal sciences and comparative medicine research in China.

Expression and significance of immune cells in patients with hepatitis B virus-related acute-on-chronic pre-liver failure / 临床肝胆病杂志

Objective To investigate the expression of myeloid-derived suppressor cells (MDSC), regulatory T cells (Treg), IL-17-producing CD4 + T cells (Th17), and CD8 + T cells (Tc17) in hepatitis B virus-related acute-on-chronic pre-liver failure (pre-ACHBLF), and to provide ideas for the early treatment of acute-on-chronic hepatitis B liver failure (ACHBLF). Methods A total of patients with pre-ACHBLF and 15 patients with ACHBLF who were hospitalized in Shijiazhuang Fifth Hospital, from August 2018 to May 2019 were enrolled as subjects, and 15 patients with chronic hepatitis B (CHB) and 15 healthy controls (HC) who underwent physical examination were enrolled as controls. Flow cytometry was used to measure the expression levels of MDSC and Th17, Treg, and Tc17 cells in peripheral blood; a blood analyzer was used to measure routine blood parameters and calculate neutrophil-to-lymphocyte ratio (NLR), monocyte-to-lymphocyte ratio (MLR), platelet-to-lymphocyte ratio (PLR), and systemic immune-inflammation index(SIRS) to evaluate the degree of inflammation, and the correlation between the expression of immune cells and the degree of inflammation was analyzed. An analysis of variance for independent samples was used for comparison of normally distributed continuous data between multiple groups, and the least significant difference t -test was used for further comparison between two groups; the Kruskal-Wallis H test was used for comparison of non-normally distributed continuous data between multiple groups, and the Nemenyi test was used for further comparison between two groups. A Pearson linear correlation analysis or Spearman's rank correlation analysis was used to investigate the correlation between variables. Results Compared with the CHB group, the ACHBLF and pre-ACHBLF groups had significant increases in the expression levels of Th17, Treg, and Tc17 cells, and the pre-ACHBLF group also had a significant increase in the expression level of MDSC (all P < 0.05). The correlation analysis showed that in pre-ACHBLF patients, MDSC were positively correlated with leukocyte count, neutrophil count, NLR, MLR, and SII ( r =0.775, 0.727, 0.571, 0.786, and 0.846, all P < 0.05), and Treg cells were only positively correlated with leukocyte count ( r =0.618, P =0.043); Th17/Treg ratio and Tc17 cells were negatively correlated with the number of lymphocytes ( r =-0.790 and -0.795, both P < 0.05). Conclusion Cellular immune dysfunction is observed in patients with pre-ACHBLF, and the expression of MDSC is closely associated with the degree of inflammation and should be taken seriously in the early stage.

Practical exploration of blended teaching in Histology and Embryology / 中华医学教育探索杂志

Blended teaching can promote the learning engagement of students and enhance their experience by combining online independent learning and offline class learning. We applied blended teaching in the course of Histology and Embryology in the large class of 147 nursing undergraduates in Chongqing Medical University. First, the teaching method was designed by a pre-course survey, and online resources were constructed. Second, students' online learning activities and group discussions were guided by a learning map. Then, flipped classes were carried out in offline class hours. Finally, an end-of-course survey and final exam scores were used evaluate the effectiveness of blended teaching. A total of 142 valid questionnaires were returned; 123 students (86.6%) approved of the effectiveness of blended teaching, 133 students (93.7%) showed improved abilities in various aspects, and 79 students (55.6%) were able to complete their online learning tasks in fewer hours than required for face-to-face lectures. The final exam results showed that the average score of blended teaching class increased by 3.5 points compared with that of the traditional face-to-face lecture class. In conclusion, blended teaching in Histology and Embryology in the large class can achieve good learning and teaching effects and thus holds promise for application.

S100A9 activates NF-κB to promote TLR7 expression and inflammatory cytokine release in microglia / 神经解剖学杂志

Objective:To investigate the effect of S100 calcium-binding protein A9(S100A9)activation of nuclear factor kappa-B(NF-κB)on the upregulation of toll-like receptor 7(TLR7)expression and the release of inflammatory factors in microglia,as well as its underlying mechanism.Methods:The viability of BV2 microglia was assessed using CCK-8 kit.Transcriptome sequencing was employed to compare differential genes(DEGs)and identify target genes from the pool of differentially expressed genes.This analysis was complemented by GO analysis,KEGG enrichment analysis and the STRING database.The expression of TLR7 mRNA was verified by real time RT-PCR.The expressions of CD68 and CD206 were detected using immunofluorescence.The expressions of CD68,CD206,TLR7,p65,and p-p65 were detected using Western Blot.The level of interleukin 6(IL-6)and tumor necrosis factor alpha(TNF-α)were verified by ELISA.Results:Moderate concentrations of S100A9 had no inhibitory effect on microglial viability.Compared to the control group,the experimental group showed a significant increase in the expression level of CD68 pro-tein,while the CD206 protein was decreased.This suggests that S100A9 promotes the activation of BV2 microglia into pro-inflammatory types.TAK-242,an inhibitor of toll-like receptor 4(TLR4),significantly inhibited the expression levels of TNF-α and IL-6 after S100A9 stimulated BV2 cells.Activation of the TLR4/NF-κB pathway promoted the ex-pression of TLR7 protein.Conclusion:The moderate concentration of S100A9 can promote the polarization of microglia towards a proinflammatory direction.It also promotes the expression of TLR7 and the release of various inflammatory factors,including TNF-α and IL-6,through the activation of the TLR4/NF-κB pathway.This activation has an obvious proinflammatory effect.

Explanation and Elaboration for the ARRIVE Guidelines 2.0—Reporting Animal Research and In Vivo Experiments (Ⅲ) / 实验动物与比较医学

Improving the reproducibility of biomedical research results is a major challenge.Researchers reporting their research process transparently and accurately can help readers evaluate the reliability of the research results and further explore the experiment by repeating it or building upon its findings. The ARRIVE 2.0 guidelines, released in 2019 by the UK National Centre for the Replacement, Refinement and Reduction of Animals in Research (NC3Rs), provide a checklist applicable to any in vivo animal research report. These guidelines aim to improve the standardization of experimental design, implementation, and reporting, as well as the reliability, repeatability, and clinical translatability of animal experimental results. The use of ARRIVE 2.0 guidelines not only enriches the details of animal experimental research reports, ensuring that information on animal experimental results is fully evaluated and utilized, but also enables readers to understand the content expressed by the author accurately and clearly, promoting the transparency and integrity of the fundamental research review process. At present, the ARRIVE 2.0 guidelines have been widely adopted by international biomedical journals. This article is a Chinese translation based on the best practices of international journals following the ARRIVE 2.0 guidelines in international journals, specifically for the complete interpretation of the ARRIVE 2.0 guidelines published in the PLoS Biology journal in 2020 (original text can be found at https://arriveguidelines.org). The third part of the article includes the items 8-10 of ARRIVE 2.0 Essential 10, which covers "experimental animals" "experimental procedures" and "results". Its aim is to promote the full understanding and use of the ARRIVE 2.0 guidelines by domestic researchers, enhance the standardization of experimental animal research and reporting, and promote the high-quality development of experimental animal technology and comparative medicine research in China.

Value of serum complement C3 in the diagnosis of liver fibrosis associated with primary biliary cholangitis / 临床肝胆病杂志

‍ ObjectiveTo investigate the value of serum complement C3 level in determining the stage of liver fibrosis in primary biliary cholangitis （PBC）. MethodsClinical data were collected from 108 patients with PBC who attended Tianjin Second People’s Hospital and underwent liver biopsy from January 2012 to October 2022. The degree of liver fibrosis （S0-4） was assessed according to the Scheuer scoring system， with ≥S2 defined as significant liver fibrosis， ≥S3 defined as progressive liver fibrosis， and S4 defined as liver cirrhosis. The independent samples t-test was used for comparison of normally distributed continuous data between two groups， and a one-way analysis of variance was used for comparison between multiple groups； the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between two groups， and the Kruskal-Wallis H test was used for comparison between multiple groups； the chi-square test or the Fisher’s exact test was used for comparison of categorical data between groups. The area under the ROC curve （AUC） was used to evaluate the efficacy of complement C3 in the diagnosis of liver fibrosis in patients with PBC. The Spearman correlation analysis was used to investigate the correlation between complement C3 and liver fibrosis stage. ResultsAmong the 108 patients with PBC， there were 87 （80.6%） female patients and 102 patients （94.4%） with positive autoantibody. As for the stage of liver fibrosis， there were 5 patients （4.6%） in S0 stage， 41 （38.0%） in S1 stage， 23 （21.3%） in S2 stage， 25 （23.1%） in S3 stage， and 14 （13.0%） in S4 stage. There was a significant difference in the level of complement C3 between the patients with different liver fibrosis stages （H=42.891， P<0.001）. The level of complement C3 gradually decreased with the aggravation of liver fibrosis， with a negative correlation between them （r=-0.565， P<0.001）. Liver stiffness measurement （LSM）， aspartate aminotransferase/alanine aminotransferase ratio， aspartate aminotransferase-to-platelet ratio index， and fibrosis-4 were negatively correlated with complement C3， with a correlation coefficient of -0.439 （P<0.001）， -0.323 （P=0.001）， -0.206 （P=0.033）， and -0.291 （P=0.002）， respectively. The multivariate logistic regression analysis showed that complement C3 level was an independent predictive factor for significant liver fibrosis， progressive liver fibrosis， and liver cirrhosis， while LSM was an independent predictive factor for significant liver fibrosis and progressive liver fibrosis. The ROC curve analysis showed that complement C3 had an AUC of 0.731， 0.832， and 0.968， respectively， in the diagnosis of significant liver fibrosis， progressive liver fibrosis， and liver cirrhosis， with a corresponding cut-off value of 1.445， 1.235， and 1.005， respectively， and complement C3 combined with LSM had an AUC of 0.811， 0.941， and 0.976， respectively， in the diagnosis of significant liver fibrosis， progressive liver fibrosis， and liver cirrhosis. There was a significant difference in AUC between complement C3 combined with LSM and complement C3 alone in the diagnosis of significant liver fibrosis （Z=2.604， P=0.009）， and there was also a significant difference in AUC between complement C3 combined with LSM and complement C3 alone in the diagnosis of progressive liver fibrosis （Z=3.033， P=0.002）； there was no significant difference in AUC between complement C3 combined with LSM and complement C3 alone in the diagnosis of liver cirrhosis （Z=1.050， P=0.294）， while There was a significant difference in AUC between complement C3 combined with LSM and LSM alone in the diagnosis of liver cirrhosis （Z=2.326， P=0.020）. ConclusionSerum complement C3 level has a certain clinical value in assessing the degree of liver fibrosis in patients with PBC， and complement C3 combined with LSM can further improve the efficacy of complement C3 or LSM in the diagnosis of liver fibrosis in PBC.

TGF-β1 promotes migration and invasion of gastric cancer cells by regulating fatty acid synthase / 中国肿瘤临床

Objective: To investigate the effect of TGF-β1 on the migration and invasion of gastric cancer cells, by regulating fatty acid synthetase. Methods: From January 2016 to December 2018, 54 paraffin samples were collected from Shunde Hospital of Southern Medical University. Immunohistochemistry was used to detect the expression of transforming growth factor β1 (TGF-β1) and fatty acid synthetase (FASN) in human gastric cancer tissues, and the correlation between them was analyzed. Transient transfection of TGF-β1 and TGF-β1 siRNA was used to construct gastric cancer cell lines. Real-time PCR was used to detect FASN mRNA level, and Western blot was performed to examine FASN protein level. Transient transfection of FASN siRNA was used to construct gastric cancer cell lines. Wound healing assays and Transwell assays were performed to observe the effect of migration and invasion ability of gastric cancer cells. Results: Immunohistochemical results indicated that the expression of TGF-β1 and FASN in gastric cancer tissue was significantly positively correlated. TGF-β1 overexpression plasmid transfection significantly up-regulated the expression level of FASN mRNA and protein, whereas TGF-β1 siRNA significantly inhibited the expression of FASN. After co-transfection with FASN siRNA, TGF-β 1-induced N-cadherin protein expression was reduced and E-cadherin protein expression was enhenced. Scratch and Transwell experiments showed that the increased cell migration and invasion by TGF-β1 were significantly decreased. Conclusions: FASN plays an important role in the migration and invasion of gastric cancer cells regulated by TGF-β1.

Analysis of the effect of mechanical thrombectomy for acute anterior circulation ischemic stroke in over-time window / 中风与神经疾病杂志

@#Objective　To analyze the therapeutic effect of mechanical thrombectomy (MT) in patients with acute anterior circulation ischemic stroke (AACIS) in over-time window.Methods　From January 2018 to November 2018,40 patients with AACIS treated with MT were enrolled in our hospital.Patients were divided into in-time-window group (onset-treatment time ≤6 h,n=30) and over-time-window group (onset-treatment time>6 h,n=10) according to the time of onset-treatment.The therapeutic effect was evaluated by modified thrombolysis in cerebral infarction (mTICI).The National Institutes of Health Stroke Scale (NIHSS) scores were compared before and after treatment to evaluate the improvement of neurological deficits.The modified Rankin scale (mRs) score,complication rate,and mortality were compared at 90 days postoperatively to assess prognosis.Results　The success rate of patients in the in-time-window group was 83.33%（25/30）,and the success rate of patients in the over-time-window group was 80.00% (16/20).There was no significant difference in the success rate between the two groups (P>0.05).The NIHSS scores at the time of discharge were lower than those at admission (P<0.05).There was no significant difference in NIHSS scores between the two groups (P>0.05).Within 90 days after surgery,no deaths occurred in either group.There was no significant difference in the prognosis rate and complication rate between the two groups (P>0.05).Conclusion　MT can effectively improve the recanalization rate of patients in the treatment of patients with AACIS over-time-window,and has similar short-term and long-term treatment effects as patients in the in-time-window.

Molecular characteristics and association analysis with litter size trait for porcine KLK7 gene.

Kallikrein-related peptidase 7(KLK7) is a tumor-related gene. In this study, the full-length coding sequence of porcine KLK7 gene was cloned through RT-PCR. Sequence analysis revealed that the pig KLK7 gene encodes a protein of 257 amino acids which has high homology with the KLK7 protein of six species: polar bear (95%), Weddell seal(94%), dog (92%), Pacific walrus (95%), domestic cat (92%), and Amur tiger (91%). This gene is structured in five exons and four introns as revealed by computer-assisted analysis. Phylogenetic analysis showed that the pig KLK7 gene has a closer genetic relationship with the KLK7 gene of a domestic cat. PCR-Alu I-RFLP was established to detect the GU373714:c.390 C > T substitution of porcine KLK7 gene and eight pig breeds displayed obvious genotype and allele frequency differences at this mutation locus. Association of this SNP with litter size trait was assessed in Large White (n = 200) and Landrace (n = 200) pig populations and results demonstrated that this polymorphic locus was significantly associated with the litter size of all parities in Large White and Landrace sows (p < 0.05). Therefore, KLK7 is also an important reproduction related gene.

Regulatory effect of mTOR pathway-mediated autophagy in liver injury / 临床肝胆病杂志

Autophagy can regulate liver physiology and balance liver metabolism. Autophagy activation has a double-sided and complex effect on liver injury, and it is regulated by many factors and is associated with many protein pathways. This article summarizes the role of mTOR in the regulation of autophagy, which can inhibit or enhance autophagy through the PI3K/Akt upstream signaling pathway and participate in the physiological and pathological changes of related liver diseases. Therefore, this article reviews the research advances in the mTOR/PI3K/Akt autophagy pathway in liver injury, in order to provide new therapeutic targets for related liver diseases.

Effects of salvianolic acid B on the migration and tube formation of high glucose-induced retinal vascular endothelial cells and the underlying mechanism / 国际中医中药杂志

Objective:To observe the effects of salvianolic acid B on migration and tube formation of the retinal vascular endothelial cell (RVEC) in high glucose, and explore its mechanism with network pharmacology.Methods:The cells were divided into normal group, model group and 1.0, 0.5, 0.1 μg/ml salvianolic acid B group according to the random number table method. The cells of each group were added with 5.5 mmol/L glucose for intervention, and the salvianolic acid B group was added with 1.0, 0.5, and 0.1 μg/ml salvianolic acid B for intervention. After 72 h, the cell viability of each group was detected by the CCK-8 method. The cells were divided into normal group, model group and low-, medium-, and high-dose salvianolic acid B group according to the random number table method. Then the cells of the normal group were added with 5.5 mmol/L glucose; the model group was added with 25 mmol/L glucose; the low-, medium-, and high-dose salvianolic acid B group was added with 25 mmol/L glucose and 0.062 5, 0.1250, 0.250 0 μg/ml salvianolic acid B. Then by taking Transwell test to detect the number of cell migration, and Matrigel test to analyze the total length of cells tubes. The active targets of Salvianolic acid B were screened by SuperTarget and Swiss TargetPrediction. Then, the targets of diabetic retinopathy were obtained by searching the GAD database, pharmGkb database, TTD database, DiGSeE database and OMIM database. The effective targets of drug-disease interaction were screened, and the component-target-disease interaction network was constructed by Cytoscape. Finally, the effective targets were analyzed by DAVID for GO analysis and KEGG pathway enrichment analysis. Molecular docking was performed by using Accelrys Discovery Studio Client 2.5 software.Results:The CCK-8 method showed that the cell absorbance values of 0.5 and 0.1 μg/ml salvianolic acid B group were not significantly different from those of the normal group ( P>0.05). The results of Transwell experiment and Matrigel experiment showed that compared with the model group, the relative number of migrating cells and the total length of tubule formation in each dose group of salvianolic acid B decreased ( P<0.05 or P<0.01). The interaction network revealed that salvianolic acid B acted on 46 targets and 8 signaling pathways. Conclusions:Salvianolic acid B could inhibit the migrating and tube forming ability of RVEC cultivated by high glucose. The results suggest that salvianolic acid B may play roles in preventing diabetic retinopathy.

Factors associated with significant liver fibrosis in chronic hepatitis B patients with non-alcoholic fatty liver disease / 中华传染病杂志

Objective:To investigate the influencing factors of significant liver fibrosis in patients with chronic hepatitis B (CHB) concurrent with non-alcoholic fatty liver disease (NAFLD).Methods:Those who underwent liver pathological examination and confirmed diagnosis of CHB and NAFLD in Tianjin Second People′s Hospital from August 2014 to September 2017 were enrolled. Data regarding their demographic information, laboratory tests results, and liver pathology results were analyzed. The latter results were used to categorize the patients either in non-significant liver fibrosis group (Metavir stage<F2) or in significant liver fibrosis group (Metavir stage≥F2). The measurement data were compared using t test or Mann-Whitney U test, and the count data using chi-square test.The factors influencing the onset of significant liver fibrosis were subsequently explored with binary logistic regressions. Results:Out of 273 patients screened, 160 and 113 patients respectively belonged to the non-significant fibrosis group and the significant fibrosis group. Age, histologic activity, NAFLD type, liver stiffness measurement, hepatitis B e antigen (HBeAg) status (positive/negative), hepatitis B virus (HBV) DNA, aspartate aminotransferase, γ-glutamyl transpeptidase, total bilirubin, high blood glucose (with/without) and platelet count between the two groups were statistically significant( t=2.232, χ2=44.276, χ2=4.808, t=2.096, χ2=5.299, t=3.191, U=7 041.500, U=6 873.500, t=2.989, χ2=5.588, t=3.429, all P<0.05). Logistic regression showed that non-alcoholic steatohepatitis (NASH), histologicactivity, HBV DNA and platelet count were the independent influencing factors for significant liver fibrosis (odds ratio ( OR)=2.809, 6.730, 0.843, 0.995, respectively, all P<0.05). Patients were divided into two subgroups according to their HBeAg status, the results showed that for patients with negative HBeAg, NASH, histologic activity, HBV DNA and platelet count were the independent influencing factors for significant liver fibrosis ( OR=8.629, 3.626, 0.740, 0.992, respectively, all P<0.05). For patients with positive HBeAg, histologic activity and high blood glucose were the independent risk factors for significant liver fibrosis ( OR=12.738, 4.223, respectively, both P<0.01). Conclusion:Liver inflammation, NASH and high blood glucose are the serious risk factors during the onset and progression of significant liver fibrosis in patients with CHB and NAFLD, while HBV DNA and platelet count levels are negatively correlated with significant liver fibrosis.

Influence of Defects at Bonding Interface on Stress Distributions in Monolithic Ceramic Crown / 医用生物力学

Objective To explore the effects of the interfacial debonding caused by water environment in the mouth and the interfacial defects between the crown and cement on stress distributions in all-ceramic crowns. Methods The three-dimensional solid model of lithium disilicate CAD/CAM crowns for the first mandibular molar was established. Seven debonding states between inferior surface of the crown and top surface of the cement (Stage 1-7) as well as two interfacial defects (Case I and II) were defined in finite element software ABAQUS. The bottom of nine models was completely constrained. For stress calculation, the 600 N vertical load was applied at occlusal surface via an analytical rigid hemisphere with the diameter of 5 mm. Results Under occlusal vertical load, the stress on interior of the crown and top surface of the cement was mainly distributed at the boundary of the debonding areas and margin of the defects. The first principle stress on interior of the crown did not exceed its ultimate tensile strength, but the maximum tensile stress of the cement exceeded its ultimate tensile strength, leading to cohesive failure in the cement. Conclusions The axial wall played a critical role in maintaining the principal tensile stress of the crown at a lower level. The defects at bonding interface between the crown and cement had a more significantly impact on load capacity of the crown than the increase in debonding areas. In order to improve load bearing capacities of all-ceramic crowns, attention should be paid to avoid defects in clinical prosthodontic practices.

Influence of Defects at Bonding Interface on Stress Distributions in Monolithic Ceramic Crown / 医用生物力学

Objective To explore the effects of the interfacial debonding caused by water environment in the mouth and the interfacial defects between the crown and cement on stress distributions in all-ceramic crowns. Methods The three-dimensional solid model of lithium disilicate CAD/CAM crowns for the first mandibular molar was established. Seven debonding states between inferior surface of the crown and top surface of the cement (Stage 1-7) as well as two interfacial defects (Case I and II) were defined in finite element software ABAQUS. The bottom of nine models was completely constrained. For stress calculation, the 600 N vertical load was applied at occlusal surface via an analytical rigid hemisphere with the diameter of 5 mm. Results Under occlusal vertical load, the stress on interior of the crown and top surface of the cement was mainly distributed at the boundary of the debonding areas and margin of the defects. The first principle stress on interior of the crown did not exceed its ultimate tensile strength, but the maximum tensile stress of the cement exceeded its ultimate tensile strength, leading to cohesive failure in the cement. Conclusions The axial wall played a critical role in maintaining the principal tensile stress of the crown at a lower level. The defects at bonding interface between the crown and cement had a more significantly impact on load capacity of the crown than the increase in debonding areas. In order to improve load bearing capacities of all-ceramic crowns, attention should be paid to avoid defects in clinical prosthodontic practices.

Application of light microscopy tissue slices and digital slicing in experimental teaching of micromorphology / 中华医学教育探索杂志

Histology teaching requires a combination of theory and experiment for a better understanding of microstructure and related functions of body.On the basis of the comparison and summary of the advantages and limitations of traditional light microscopy tissue slices and the emerging digital slicing,we combined them in the teaching of micromorphology experiments to achieve a better teaching results.Each experimental course (about 3 to 4 hours) was divided into four parts:teaching videos,observation of light microscopy tissue slices and digital slicing,discussion on course content and random quizzes.This teaching method contributed to the improvement of the students' interest and motivation in learning,the teachers' teaching efficiency and the overall teaching quality.

The role of intrahepatic regulatory T cells/helper thymphorytes 17 balance in patients with different phases of chronic hepatitis B virus infection / 中华传染病杂志

Objective To analyze the change of intrahepatic regulatory T cells (Treg )/helper thymphorytes (Th)17 balance in patients with different phases of chronic hepatitis B virus (HBV ) infection ,and to explore the role of Treg/Th17 balance in maintaining immune tolerance and inducing immune clearance ,and its influence on disease progression .Methods Sixty-eight patients with chronic HBV infection who underwent liver biopsy in Tianjin Second People′s Hospital were included .The 68 patients included 20 cases in immune tolerant (IT) phase ,36 cases in immune clearance (IC) phase and 12 cases in inactive phase .Eight healthy liver transplant donors were collected as healthy controls .The intrahepatic Treg/Th17 levels were detected by immuno-histochemical method . The changes of Treg/Th17 balance in patients with different phases of chronic HBV infection ,and the relationship between Treg/Th17 balance and the decreases of hepatitis B surface antigen (HBsAg ) , hepatitis B antigen (HBeAg) and HBV DNA levels in the peripheral blood were analyzed in patients with IC phase at two weeks of admission .Results The intrahepatic Treg and Th17 levels in IC phase group were the highest , then and they were higher in inactive phase group were higher than those in IT phase group ,And they were the lowest in control group .The Treg level in IC phase group increased significantly compared with the other three groups (all P< 0 .01) ,and there were no significant differences among the other three groups (all P> 0 .05) .The Th17 level between IT phase group and inactive phase group was not significantly different (P> 0 .05) ,while the differences were not significant in other groups (all P>0 .05) .Treg/Th17 ratio of IT phase group was the highest ,then the ratio of control group was higher than that of inactive phase group ,and IC phase group was the lowest ratio .The differences between IC phase group ,control group and IT phase group were significant (all P< 0 .05) ,and the difference between inactive phase group and IT phase group was also significant (all P<0 .05);and there was no significant difference among other groups (all P>0 .05) .The decreases of HBsAg ,HBeAg and HBV DNA levels in the peripheral blood at two weeks admission were negatively correlated with the intrahepatic Treg cell level in patients in IC phase of chronic HBV infection ( r= -0 .941 ,-0 .869 ,and -0 .883 ,respectively ,both P<0 .01) .The Treg ,Th17 levels and their ratio in IC phase group with different degree of inflammation and fibrosis had significant differences :G4 group > G3 group > G2 group ,S3 group > S2 group > S1 group (all P<0 .05) .Conclusions There is no change of the Treg/Th17 balance in IT phase ,and Treg has no influence on maintaining immune tolerance in chronic HBV infection .T he imbalance of Treg/Th17 is observed in IC phase .Th17 may actively participate in the immune-mediated liver injury and the development of hepatic fibrosis in CHB patients .Treg may inhibit inflammation and reduce liver injury via the negative feedback regulation mechanism ,and may impede the eradication of HBV simultaneously .

Isolation and content determination of taxifolin, orobol and quercetin in Cudrania tricuspidata / 国际中医中药杂志

Objective To isolate and identify 3 flavonoids (taxifolin, orobol and quercetin) from Cudrania tricuspidata, and develop a method for determining 3 flavonoid constituents in Cudrania tricuspidata. Methods Three flavonoids was isolated from ethanol extract of Cudrania tricuspidata by chromatography, and its structure was identified by nuclear magnetic resonance. The analysis was conducted on an Aglient C18 column (4.6 mm ×250 mm, 5 μm) eluted with 1％ acetic acid and methanol as mobile phases in gradient mode. The flow rate was 1 ml/min and the detection wavelength was set at 310 nm. The column temperature was 25 ℃. Results Taxifolin, orobol and quercetin were isolated from ethanol extract of Cudrania tricuspidata by chromatography. The content of taxifolin, orobol and quercetin were 0.850 mg/g, 0.518 mg/g, 0.103 mg/g. Conclusion The method can be used for the quality control of Cudrania tricuspidata as a reference.

Efficacy of High-dose Morphine with Different Continuous Administration Routes:a Prospective Study in Pa-tients with Advanced Cancer Pain / 中国药师

Objective:To compare the efficacy of morphine with intravenous injection and subcutaneous injection in the treatment of advanced cancer,and explore the indications of different drug delivery methods for high-dose morphine. Methods: A prospective study was performed,and 46 cases of patients with advanced cancer pain were collected and divided into intravenous group and subcutaneous group according to the administration route. Pain was assessed during the administration,and the analgesic efficiency and the incidence of adverse reactions were observed to compare the efficacy and safety of two different ways to give high-dose morphine. Results:No statisti-cally significant differences were found in the number of outbreaks needed to be rescued,the frequency of morphine-induced drug deliver-y,the efficiency of analgesia after opioid transfer,and the incidence of opioid-related side effects between the groups (P>0.05). The dose of morphine in the subcutaneous group was higher than that of the intravenous group(P<0.05). Conclusion:The continuous ad-ministration of morphine with intravenous injection and subcutaneous injection can quickly,safely and effectively relieve pain. With the same analgesic efficacy,patients can choose appropriate administration route according to the dose of morphine, the influence degree of primary diseases and the individual will.

Study on the alkaloids in the different parts of Aconitum paniculigerum Nakai var.wulingense (Nakai) by HPLC-MS / 国际中医中药杂志

Objective To analyze the alkaloids in the different parts of Aconitum wulingense by HPLC-ESI-Trap-MS. Methods The Agilent XDB-C18(250 mm×4.6 mm, 5 μm) column with gradient elution of 0.1% solvent (A)-acetonitrile(B), at a flow rate of 1 ml/min was used. The column temperature was set at 30 ℃. The MS analysis was based on positive ions mode. Results In the roots, a total of 61 diterpenoid alkaloids were discovered, among which 46 were identified. In the stems, 38 alkaloids have been found, among which 33 alkaloids were identified and 27 were the same with the roots. In the leaves, 18 alkaloids have been detected and 8 were the same with the roots. Conclusions The method is accurate, reliable and efficient, and is suitable for rapid identification of ingredients in Aconitum wulingense, which provides a reference for the development and utilization of Aconitum wulingense and clarify its efficacy and material basis.

Optimization of preparation of Haoqin-Huaban granules / 国际中医中药杂志

Objective@#To optimize the preparation process for Haoqin-Huaban granules so as to provide experimental basis for the development and utilization of the compound.@*Methods@#Taking the extraction rate of gentiopicroside from Gentiana macrophylla Pall as the observation index, L9(34)orthogonal test was used to investigate the amount of water, extraction time, and extraction frequency for optimizing the water extracting technology of the Haoqin-Huaban granules. Then the dry paste was used for the raw material, the ratio of the diluent (dextrin) and the concentration and dosage of the binder (ethanol) were investigated by single factor, and the particles were prepared.@*Results@#The optimum water extraction process was A2B2C3, which indicated that the prescribed medicinal materials with 8 times water were extracted 1 h for 3 times. The extract was concentrated into paste, vacuum drying, pulverizing, mixing according to the ratio of 5:1, with ethanol, made of soft material, pelletizing, drying at 60 ℃ for granulation, in order to make Haoqin-Huaban granules.@*Conclusions@#The preparation process is reasonable and feasible, which provides experimental basis for the industrial production of Haoqin-Huaban granules.