Animal experiments are crucial in evaluating the preclinical safety and efficacy of new dental materials, drugs, instruments, and equipment by identifying and eliminating potential health risks to humans. An international team of several dental experts formulated a guideline named Preferred Reporting Items for Animal Studiesin Endodontology (PRIASE) 2021. Consisting of 11 domains, 43 individual items, and a flowchart. PRIASE provides guidance for animal experiments in dentistry and improves the quality of experiment design and reporting. This work introduces the process and basic content of the guideline and interprets the key items of its checklist with specific examples to provide reference for the reporting of animal experiment in dentistry in China.
Animals ; Animal Experimentation/standards* ; Checklist ; China ; Endodontics ; Guidelines as Topic ; Research Design

OBJECTIVE To provide reference for clarifying improvement effects of Anemarrhena asphodeloides on sepsis- induced myocardial injury and potential material basis. METHODS Water extract of A. asphodeloides was extracted by thermal reflux method. Total xanthone and total saponins in A. asphodeloides were separated by macroporous adsorption resin. The mice model of sepsis-induced myocardial injury was established by intraperitoneal injection of lipopolysaccharide. The effects of the location of three extraction fractions and the monomers of A. asphodeloides as mangiferin， timosaponin AⅢ and timosaponin BⅡ on the survival rate of the model mice were explored. HE staining was used to observe the effects of mangiferin， timosaponin AⅢ and timosaponin BⅡ on myocardial morphology in model mice. The effects of mangiferin on mRNA expressions of inflammatory cytokines [interleukin-6 （IL-6）， IL-1β and tumor necrosis factor-α （TNF-α）] and the level of reactive oxygen species （ROS） in myocardial tissue of model mice were detected. RESULTS Compared with the model group， the survival rate of mice in the intervention group of total xanthone， total saponins and water extract was increased to different extents， especially total xanthone fraction. Mangiferin， timosaponin AⅢ and timosaponin BⅡcould improve the degree of myocardial cell swelling and muscle bundle arrangement disorder in model mice， especially mangiferin. Compared with model group， mRNA expressions of IL-6， IL- 1β and TNF- α， ROS level in myocardium of mice after mangiferin intervention were decreased to different extents. CONCLUSIONS The different extraction fractions of A. asphodeloides can improve survival rate of mice with sepsis-induced myocardial injury， especially total xanthone fraction. Mangiferin is the best among the three monomers of A. asphodeloides to improve sepsis-induced myocardial injury， which may play a role in anti-sepsis myocardial injury by anti-inflammation and antioxidantion.

By reviewing ancient materia medica， medical and prescription books， combined with modern literature， the textual research of Stephaniae Tetrandrae Radix has been conducted to verify the name， origin， producing area， harvesting and processing methods. Through textual research， the results show that the mainstream name of this herb recorded in the past dynasties is Fangji， which is also called Hanzhong Fangji because it is produced in Hanzhong city， and after the Tang dynasty， it was gradually divided into Hanfangji and Mufangji， and there is the saying that Han Zhushuiqi， Mu Zhufengqi. The names of Fenfangji and Guangfangji were first seen in the republic of China. In addition， Fenfangji was once distributed in Hankou， so it was also known as "Hanfangji"， which is easily confused with the traditional Hanzhong Fangji for short. Based on the original research， it is concluded that Aristolochia heterophylla（Hanzhong Fangji）is the mainstream of Stephaniae Tetrandrae Radix used in the Qing dynasty and before， and the application history of Cocculus orbiculatus can be traced back to before the Tang dynasty. After the Ming dynasty， Stephania tetrandra gradually became another main origin， and in the Republic of China， A. fangchi was used as a medicine for Stephaniae Tetrandrae Radix， but in modern times it was banned because it contained aristolochic acid as a toxic ingredient， and S. tetrandra has become the mainstream legal origin. The traditional production area of Hanzhong Fangji is Hanzhong， Shaanxi province， while today the mainstream of S. tetrandra is manly produced in Jiangxi and other places. Based on the quality evaluation research， the quality of Hanzhong Fangji is better with the radial texture of section used as radial solution， yellow solid and fragrant. Fenfangji with solid quality， white inside， powdered enough， less fiber and radiating texture is better. From the harvesting and processing research， the root of Fangji is mostly harvested in spring and autumn， and the outer bark should be removed in some literature. Before the Ming dynasty， this herb was dried in the shade， and after the Ming dynasty， it was dried in the sun. The modern production processing of Fangji is to harvest it in autumn， wash it， remove the rough bark， dry it to half dry， cut it into sections， and then cut it longitudinally if it is large， and dry it. Based on the results， combined with current studies on the toxicity of aristolochic acid and influencing factors such as commercial circulation， it is suggested that S. tetrandra should be used as the origin of Fangji， the processed products are selected according to the prescription requirements， and those without specified requirements can be processed by referring to the raw products in the 2020 edition of Chinese Pharmacopoeia.

OBJECTIVE To establish the fingerprints of c ultivated and wild Anemarrhena asphodeloides，and to identify their differential components. METHODS Using an evaporative light-scattering detector ， the high performance liquid chromatography combined with Similarity Evaluation System of TCM Chromatographic Fingerprint （2012 edition） were used to establish fingerprints of 14 batches of cultivated A. asphodeloides and 14 batches of wild medicinal materials ，and evaluate their similarity. The common peaks were identified by comparison with the chromatogram of the mixed control. At the same time ，the contents of components corresponding to common peaks in cultivated and wild A. asphodeloides were determined. The principal component analysis and orthogonal partial least squares discrimination analysis were adopted to identify differential components of them ，and compare the contents of them. RESULTS Among 28 batches of A. asphodeloides ，10 common peaks were found ，i.e. neomangiferin（peak 1），mangiferin（peak 2），isomangiferin（peak 3），timosaponin B Ⅱ（peak 7），timosaponin B Ⅲ（peak 8）， timosaponin Ⅰ（peak 9），timosaponin A Ⅲ（peak 10）. The similarities of fingerprints of samples with control fingerprint were no less than 0.963. The average total contents of seven components in cultivated and wild A. asphodeloides were 74.18 and 84.72 mg/g， respectively；there was statistical significance （P＜0.05）. The cultivated and wild A. asphodeloides could be divided into two categories. The differential components were neomangiferin ，mangiferin，timosaponin B Ⅱ and timosaponin A Ⅲ（VIP values were all higher than 1）. The content of neomangiferin in cultivated products was significantly higher than that in wild products （P＜ 0.05），and the contents of mangiferin ，timosaponin B Ⅱ and ti mosaponin A Ⅲ were significantly lower than those in wild products （P＜0.05）. CONCLUSIONS Fingerprint of A. asphodeloides is established ，and differential components of cultivated and wild A. asphodeloides are identified primarily.

Objective:To explore and evaluate a appropriate suitable method for detection of Campylobacter and antibiotic sensitivity test for foodborne diarrhea in clinical laboratories. Methods:Pre-experiment:a total number of 400 fecal samples of patients with foodborne diarrhea were prospectively collected from the intestinal disease clinic of Beijing Tongren Hospital from September 2017 to January 2018. Double-hole filtration culture method and modified cefoperazone charcoal deoxycholate (CCD) agar culture method were used for fecal culture in micro-aerobic environment for 48 hours, and then suspicious colonies were identified by matrix assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry. Meanwhile, C. jejuni and C. coli were detected by real-time quantitative polymerase chain reaction(qPCR). Large sample verification: 2 062 fecal samples of patients with foodborne diarrhea in three hospitals of different levels in different areas of Beijing were collected for qPCR detection and culture from April 2018 to March 2019. The antimicrobial sensitivity test (AST) of C. jejuni and C. coli was performed according to the disk diffusion method and agar dilution method recommended by Clinical and Laboratory Standards Institute and National Antimicrobial Resistance Monitoring System for Enteric Bacteria. The results of the three detection methods and the consistency of the two antibiotic sensitivity tests were compared. Results:In the pre-experiment, the positive rates of Campylobacter ( jejuni/coli) detected of qPCR, double-hole filtration culture and modified CCD agar culture were 9.0% (36/400), 5.0% (20/400)and 3.5% (14/400), and the difference was statistically significant ( P<0.01). The samples with negative result of qPCR were negative by both culture methods. The total positive rates of Campylobacter detected by qPCR was 8.1% (168/ 2 062)including 7.0% (144/2 062) for C. jejuni and 1.2% (24/2 062) for C. coli. The samples with positive qPCR results were cultured by double-hole filtration culture method and the positive rate was 61.9%(104/168), among which, the positive rate of C. jejuni and C. coli were 58.3%(84/144) and 83.3%(20/24) respectively, which was not significantly different from the detection rate and culture positive rate in the pre-test ( P>0.1). The resistance rates of C. jejuni and C. coli to ciprofloxacin were 94.0%(94/100) and 100.0%(24/24) and to erythromycin were 6.0%(6/100) and 33.3%(8/24). The results from two antibiotic sensitivity test methods were consistent (Kappa>0.75). Conclusions:qPCR is rapid, sensitive and easy to operate, so it is suitable for routine development in clinical laboratories. The double-hole filtration culture method is beneficial to the acquisition of strains and is essential for the further study of Campylobacter. There was no significant difference between agar dilution method and disk diffusion method in antibiotic sensitivity test. Campylobacter showed a very high resistance rate to quinolones, which was no longer suitable for the treatment of Campylobacter foodborne diarrhea in Beijing area. Macrocyclic lipid antibiotics should be preferred.

The clinical characteristics, mutation analysis results, and family tree of a patient with autosomal dominant Alport syndrome (ADAS), who had nephrotic syndrome as the first manifestation were examined.The proband was a 11-month-old girl who presented with nephrotic syndromes and gross hematuria.During the treatment course, the patient had steroid resistance and a poor response to Cyclosporine and Cyclophosphamide pulse therapy.Renal biopsy was performed 2 years after disease onset.Under the light microscopy, glomerular segmental mesangio-proliferative lesions were observed.The staining of type Ⅳ collagen showed the loss of the α3 chain in the glomerular basement membrane (GBM) and tubular basement membrane, and α5 chain loss in GBM.Electron microscopy showed uneven thickness of GBM, with obviously delaminated and tearing dense basement membrane (BM) layer, showing a typical lace-like change.The segmental BM was loosened and widened.Her father did not develop microscopic hematuria until 10 years later, while her grandmother had asymptomatic hematuria and proteinuria when the proband was diagnosed.A new mutation in the COL4A4 gene was found in the proband, namely c. 1715delG (p.G572Vfs * 81). Her father and grandmother carried the same mutation, but her mother and sister did not have.The clinical manifestation of ADAS is clinically heterogeneous and its incidence may be higher than what we have expected.

Objective To analyze the correlation between urinary albumin/creatinine ratio (ACR) and 24-hour urinary microalbumin (UMA) and evaluate the predictive value of ARC for early diabetic nephropathy.Methods A total of 368 patients with type 2 diabetes mellitus were retrospectively collected.Early diabetic nephropathy was defined as 24h UMA 30～＜300 mg/24h.The correlation between ACR and 24hUMA,and the area under the receiver operating characteristic (ROC) curve of ACR in diagnosis of early diabetic nephropathy were calculated.Gender,age,course of disease,fasting venous blood glucose,glycosylated hemoglobin,blood pressure,triglyceride and total cholesterol were used as adjusting variables to establish univariate and multivariate logistic models of ACR for early diabetic nephropathy,respectively.A regression model was used to evaluate the diagnostic value of ACR for early diabetic nephropathy.Results The correlation between ACR and 24h UMA was 0.658.The area under ROC curve of ACR for early diabetic nephropathy was 0.907 before and 0.933 after adjustments of gender,age,course of disease,fasting venous blood glucose,glycosylated hemoglobin,blood pressure,triglyceride and total cholesterol,respectively.The OR value of ACR of diabetic nephropathy was 2.016 before and 2.762 after same adjustments.The calibration of Hosmer-Lemeshow chi-square test evaluation model was 19.362 before (P=0.13) and 14.928 after adjustments (P=0.061).Conclusion ACR is a better predictor for early diabetic nephropathy although its value is influenced by gender,age,course of disease,blood sugar,lipid,and blood pressure.

Objective:To investigate the expression and clinical significance of PD-L1 in colorectal cancer (CRC). Methods:A total of 210 CRC patients who accepted radical surgery in our hospital from January 2015 to January 2016 were divided into three groups, namely, high-frequency microsatellite instability (MSI-H), low-frequency microsatellite instability (MSI-L), and microsatellite stable (MSS). The expression of PD-L1 was detected by immunohistochemistry, and the expression characteristics of PD-L1 in different types of CRC were analyzed. Results:CRC cases with low differentiation had a higher expression of PD-L1 than CRC patients with high differ-entiation (P<0.05). PD-L1 had a positive rate of 75.8%in the MSI-H group and a rate of 9.3%in the MSI-L and MSS groups, wherein the difference between the two groups was statistically significant (P<0.05). Conclusion:PD-L1 was positively expressed in some CRC tu-mor tissues, and its positive rate was significantly higher in MSI-H than in MSI-L and MSS. The therapeutic effect of a PD-L1 blocker for patients with MSI-H CRC might be preferable.

Objective To establish the finite element model of uterosacral ligament (USL) and cardinal ligament (CL) and analyze the stress distribution and deformation with USL and CL under different working conditions. Methods Patients with stage Ⅲ-Ⅳpelvic organ prolapse (POP) and healthy female volunteers were selected for research subject, and divided into anterior uterus group and posterior uterus group. Two POP patients and two volunteers were selectd into the anterior uterus group and posterior uterine group respectively. Pelvic MRI scan was performed in two groups. Based on the original MRI data sets, the finite element model of USL and CL was constructed by using the software such as the Mimics, and the stress distribution and deformation of USL and CL were simulated. Results Under the premise of the elastic modulus fixed and three different working conditions such as 60 cmH2O, 99 cmH2O and 168 cmH2O (1 cmH2O=0.098 kPa) with abdominal pressure generated by maximum Valsalva maneuver, according to the present conditions and the simulation, the trend was analyzed: the stress and deformation of the uterus, anterior vaginal wall, USL and CL in two groups were mainly distributed in the middle and lower part of the anterior vaginal wall or the ligament and the cervix-vagina junction, the maximum stress and the maximum displacement were mainly concentrated in the lower region of the anterior vaginal wall. With increasing of abdominal pressure generated by the maximum Valsalva maneuver, the maximum stress values of the POP patient in anterior uterus group under three different working conditions were: 0.027 9, 0.046 0, 0.078 0 MPa, and the maximum displacement values were: 9.145 5, 15.090 0, 25.607 0 mm. The maximum stress values of the volunteer in anterior uterus group under three different working conditions were:0.012 6, 0.020 8, 0.035 3 MPa, and the maximum displacement values were: 1.816 7, 2.997 5, 5.086 7 mm. The maximum stress values of the POP patient in posterior uterine group under three different conditions were: 0.069 4, 0.114 6, 0.194 5 MPa, and the maximum displacement values were:11.658 0, 19.236 0, 32.643 0 mm. The maximum stress values of the volunteer in posterior uterus group under three different working conditions were:0.009 1, 0.015 1, 0.025 6 MPa, and the maximum displacement values were:2.581 6, 4.259 6, 7.228 4 mm. The maximum stress values and the maximum displacement values were all increased with increasing of abdominal pressure in the two groups. The maximum stress values and the maximum displacement values of the POP patients were greater than those of volunteers. Under different working conditions, the maximum stress values and maximum displacement values of the posterior uterus POP patient were all greater than those of the anterior uterus POP patient. Conclusions The finite element model of USL and CL is completely based on the MRI technology and the model is real and reliable. The increase of abdominal pressure will produce a larger stress and deformation of USL and CL, which is one of the reasons causing the injury of the ligament.