Drug-induced liver injury （DILI） is the main cause of failures in drug development and the withdrawal of approved drugs from the market， and therefore， there is an increasing demand for accurate prediction and in vitro testing. However， the two-dimensional cell culture system of hepatocytes is not suitable for the toxicity study of long-term drug use due to the fact that it cannot accurately simulate and reproduce the real environment and micro-ecosystem of hepatocytes in vivo. In view of this， there is an urgent need for liver models with higher predictability to assess the hepatotoxicity of drugs in drug development and the safety evaluation of active compounds. This article reviews the construction and application of three-dimensional in vitro hepatocyte culture systems for DILI， in order to provide a reference for their effective implementation in DILI analysis.

Objective: To investigate the potential mechanism of Wendan decoction in obesity by screening target genes with promoter region methylation changes and constructing a multiple signaling pathways network based on promoter methylation. Methods: The methylation degree of Itgad, Col8a1, Adra2b, Jund, Rab2a, Wnt8b, Fzd9, B4galt7, Pik3cd, Creb1, Stard8, and Mmp1 in the abdominal adipose tissue of obese rats was determined using the Agena MassARRAY system. Western blot was performed to assess protein expression levels. Target genes were identified based on the methylation degree in the promoter region and protein expression. Enrichment analysis of signaling pathways was conducted to identify relevant target genes and obtain a multiple signaling pathway network associated with obesity. Core and terminal effector molecules in the pathway networks were selected as research targets for reverse transcription-polymerase chain reaction (RT-PCR) analysis. Results: Four genes (Adra2b, Creb1, Itgad, and Pik3cd) showed a degree of promoter methylation consistent with their respective protein expression levels. Among them, Adra2b, Creb1, and Pik3cd expression increased, while that of Itgad decreased. Enrichment analysis revealed that Creb1 and Pik3cd were involved in 6 signaling pathways related to obesity: tumor necrosis factor (TNF) signaling pathway, growth hormone synthesis/secretion and action, adenosine 5′-monophosphate-activated protein kinase (AMPK) signaling pathway, relaxin signaling pathway, cyclic nucleotide (cAMP) signaling pathway, and phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt) signaling pathway. Subsequently, a multiple signaling pathways network was constructed based on promoter methylation. Key molecules including protein kinase B (AKT), mechanistic target of rapamycin complex 1 (mTORC1), and unc-51 like autophagy activating kinase 1 (ULK1), as well as terminal effector molecules interleukin-1β (IL-1β), interleukin-6 (IL-6), and chemokine (C-X-C motif) ligand 2 (CXCL2) were selected as research targets. Wendan decoction decreased the expressions of AKT, mTORC1, IL-1β, IL-6, and CXCL2 while up-regulating ULK1 expression. Conclusion The mechanism of Wendan decoction in preventing obesity involves the regulation of multiple signaling pathways through the control of Creb1 and Pik3cd gene promoter methylation. However, the associated multi-path gene regulation mechanism in preventing obesity is complex. Thus, further exploration is needed to elucidate the role of methylation changes in this mechanism.

Objective:To investigate the diagnostic performance of CT-derived fractional flow reserve (CT-FFR) derived from standard images (STD), images processed by first-generation (SSF1) and second-generation (SSF2) whole-heart motion correction algorithm, respectively.Methods:Patients who underwent both coronary CT angiography (CCTA) and invasive coronary angiography (ICA) with FFR examination within 3 months in Shanghai General Hospital, Shanghai Jiao Tong Univerisity School of Medicine from January 2020 to December 2022 were screened in this retrospective study. Totally of 121 patients (134 lesions) were finally included in the study. CCTA images were reconstructed using iterative reconstruction, iterative reconstruction plus SSF1 and SSF2 algorithms. All images were divided into three groups: STD group, SSF1 group, and SSF2 group. The image quality of the CCTA images was assessed using the Likert scale, and differences between the two groups were compared using the Mann-Whitney U and Kruskal-Wallis test. The correlation and consistency between CT-FFR and FFR were evaluated using Spearman correlation coefficient and Bland-Altman plots. The diagnostic performance of CCTA and CT-FFR from three groups was compared by receiver operating characteristic (ROC) curves. The area under the curve (AUC) was compared using the DeLong test. Results:Compared to the STD group and SSF1 group, the SSF2 group showed the best performance in image quality score (median=3.7). Best correlation ( r=0.652, P<0.001) and consistency (mean difference=0.03) between CT-FFR and FFR were observed in SSF2 group. ROC analysis results revealed that, at the per-lesion level, in the diagnosis of ischemic lesions, the diagnostic performance of CT-FFR in the SSF2 group was significantly better than that of the SSF1 group (AUC=0.88 vs. 0.76, P=0.003), while no significant difference was observed between STD group and SSF1 group ( P=0.125). At the per-patient level, the SSF2 group also demonstrated the highest diagnostic performance. Conclusion:The SSF2 algorithm significantly improved CCTA image quality and enhanced its diagnostic performance for evaluating stenosis severity and CT-FFR calculations.

Objective:To investigate the impact of lifestyle, apolipoprotein E (ApoE) gene, and their interaction on the risk for cognitive frailty in the elderly population in China.Methods:The study participants were from the Chinese Longitudinal Healthy Longevity Survey. The information about their lifestyles were collected by questionnaire survey, and a weighted lifestyle score was constructed based on β coefficients associated with specific lifestyles to assess the combined lifestyle. ApoE genotypes were assessed by rs429358 and rs7412 single nucleotide polymorphisms. Cognitive frailty was assessed based on cognitive function and physical frailty. Cox proportional hazards regression model was used to analyze the association of lifestyle and ApoE gene with the risk for cognitive frailty and evaluate the multiplicative and additive interactions between lifestyle and ApoE gene. Results:A total of 5 676 elderly persons, with median age [ M ( Q1, Q3)] of 76 (68, 85) years, were included, in whom 615 had cognitive frailty. The analysis by Cox proportional hazards regression model indicated that moderate and high levels of dietary diversity could reduce the risk for cognitive frailty by 18% [hazard ratio ( HR)=0.82, 95% CI: 0.68-1.00] and 28% ( HR=0.72, 95% CI: 0.57-0.91), respectively; moderate and high levels of physical activity could reduce the risk by 31% ( HR=0.69, 95% CI: 0.56-0.85) and 23% ( HR=0.77, 95% CI: 0.64-0.93), respectively. Healthy lifestyle was associated with a 40% reduced risk for cognitive frailty ( HR=0.60, 95% CI: 0.46-0.78). ApoE ε4 allele was associated with a 26% increased risk for cognitive frailty ( HR=1.26, 95% CI: 1.02-1.56). No multiplicative or additive interactions were found between lifestyle and ApoE gene. Conclusions:Dietary diversity and regular physical activity have protective effects against cognitive frailty in elderly population. Healthy lifestyle can reduce the risk for cognitive frailty in elderly population regardless of ApoE ε4 allele carriage status.

Objective:To investigate the influencing factors of recurrence after radical resection of middle and low rectal cancer, and to establish a prediction model based on magnetic resonance imaging (MRI) measurement of perirectal fat content and investigate its application value.Methods:The retrospective cohort study was constructed. The clinicopathological data of 254 patients with middle and low rectal cancer who were admitted to Tianjin Union Medical Center from December 2016 to December 2021 were collected. There were 188 males and 66 females, aged (61±9)years. All patients underwent radical resection of rectal cancer and routine pelvic MRI examina-tion. Observation indicators: (1) follow-up and quantitative measurement of perirectal fat content; (2) factors influencing tumor recurrence after radical resection of middle and low rectal cancer; (3) construction and evaluation of the nomogram prediction model of tumor recurrence after radical resection of middle and low rectal cancer. Measurement data with normal distribution were represented as Mean± SD, and measurement data with skewed distribution were represented as M(rang) and M( Q1, Q2). Count data were described as absolute numbers. Univariate and multivariate analyses were conducted using the COX regression model. The rms software package (4.1.3 version) was used to construct the nomogram and calibration curve. The survival software package (4.1.3 version) was used to calculate the C-index. The ggDCA software package (4.1.3 version) was used for decision curve analysis. Results:(1) Follow-up and quantitative measurement of perirectal fat content. All 254 patients were followed up for 41.0(range, 1.0?59.0)months after surgery. During the follow-up period, there were 81 patients undergoing tumor recurrence with the time to tumor recurrence as 15.0(range, 1.0?43.0)months, and there were 173 patients without tumor recurrence. The preoperative rectal mesangial fascia envelope volume, preoperative rectal mesangial fat area, preoperative rectal posterior mesangial thickness were 159.1(68.6,266.5)cm3, 17.0(5.1,34.4)cm2, 1.2(0.4,3.2)cm in the 81 patients with tumor recurrence, and 178.5(100.1,310.1)cm3, 19.8(5.3,40.2)cm2 and 1.6(0.3,3.7)cm in the 173 patients without tumor recurrence. (2) Factors influencing tumor recurrence after radical resection of middle and low rectal cancer. Results of multivariate analysis showed that poorly differentiated tumor, tumor pathological N staging as N1?N2 stage, rectal posterior mesangial thickness ≤1.43 cm, magnetic resonance extra mural vascular invasion, tumor invasion surrounding structures were independent risk factors of tumor recurrence after radical resection of middle and low rectal cancer ( hazard ratio=1.64, 2.20, 3.19, 1.69, 4.20, 95% confidence interval as 1.03?2.61, 1.29?3.74, 1.78?5.71, 1.02?2.81, 2.05?8.63, P<0.05). (3) Construction and evaluation of the nomogram prediction model of tumor recurrence after radical resection of middle and low rectal cancer. Based on the results of multivariate analysis, the tumor differentiation, tumor pathological N staging, rectal posterior mesangial thickness, magnetic resonance extra mural vascular invasion, tumor invasion surrounding structures were included to construct the nomogram predic-tion model of tumor recurrence after radical resection of middle and low rectal cancer. The total score of these index in the nomogram prediction model corresponded to the probability of post-operative tumor recurrence. The C-index of the nomogram was 0.80, indicating that the prediction model with good prediction accuracy. Results of calibration curve showed that the nomogram prediction model with good prediction ability. Results of decision curve showed that the prediction probability threshold range was wide when the nomogram prediction model had obvious net benefit rate, and the model had good clinical practicability. Conclusions:Poorly differentiated tumor, tumor pathological N staging as N1?N2 stage, rectal posterior mesangial thickness ≤1.43 cm, magnetic resonance extra mural vascular invasion, tumor invasion surrounding structures are independent risk factors of tumor recurrence after radical resection of middle and low rectal cancer. Nomogram prediction model based on MRI measurement of perirectal fat content can effectively predict the probability of postoperative tumor recurrence.

Objective: The aim of this study was to investigate the effect of changes in nasal tip protrusion on the sense of upper lip protrusion in different populations and to provide a reference for the improvement of soft tissue beauty. Methods : Informed consent and portrait authorization were obtained from the model. A female model whose face met the criteria was selected, and a 3D model of her was obtained using a 3D stereo camera. Based on the original model, ZBRUSH2019 software was used to simulate changes in nose tip protrusion. Then, 9 segments of facial dynamic rotation videos were produced, and an electronic questionnaire was created through a questionnaire website to evaluate the effect of different nose tip protrusions on perceived upper lip protrusion. Randomly selected orthodontic patients, orthodontics professionals and general adults completed the electronic questionnaire. In the questionnaire, a Visual Analog Scale was used to evaluate the perceived degree of protrusion of the upper lip. The higher the score, the more prominent the upper lip of the model in the image. The questionnaire data were statistically analyzed using a generalized linear mixed model. Results : As the nasal tip position became progressively more retracted, the subjects' upper lip protrusion scores increased. Among male subjects, the results showed that the general population thought that the upper lip protrusion was higher than the patients did when the nasal tip protrusion was +6 mm and +4.5 mm (P = 0.023, P = 0.047). When the nasal tip protrusion was +6 mm, the scores of the general population were higher than those of the orthodontics professionals (P = 0.023). However, when the nasal tip variable was -6 mm, their score was lower than that of the patients (P = 0.003), and there was no significant difference in other retest distances between groups (P>0.05). Conclusion When the protrusion of the nasal tip decreased, the three groups experienced a visual illusion of upper lip protrusion. When the nasal tip protrusion is too large, the general population perceived the visual illusion of the upper lip protrusion as being more obvious.

Objective:To seek any effect of extracorporeal shockwave treatment on the expression of transforming growth factor-β1 (TGF-β1) and interleukin-1β (IL-1β) in the cartilage tissue of rabbits with knee osteoarthritis (OA), and its therapeutic mechanism.Methods:Fifty female New Zealand rabbits were randomly divided into a normal control group, a model group, and three shockwave groups A, B and C, each of 10. Except for the normal control group, an OA model was established in the other groups using Hulth′s method. The shockwave groups were given 2000 shocks in each weekly session over 4 weeks. The energy flow density in group A was 0.05mJ/mm 2; in B it was 0.11mJ/mm 2 and in C 0.22mJ/mm 2. The normal control and model groups were not shocked. All the rabbits were then sacrificed and their right knee cartilage tissue was sampled to observe any pathological changes and assign improved Mankin scores. Immunohistochemistry was used to count the number of TGF-β1 and IL-1β-positive cells in the cartilage. Western blotting and real-time fluorescence quantitative polymerase chain reactions were employed to determine the protein and mRNA expression of IL-1β and TGF-β1. Results:Compared with the normal group, degeneration of articular cartilage was observed in the model group. The average Mankin′s score of the model group was significantly higher than that of the normal control group. The average expression of TGF-β1 and IL-1β protein and mRNA in the model group had increased significantly compared with the normal control group. The average Mankin′s scores of the shock wave groups were all significantly lower than the model group′s average. Group C′s average expression levels of TGF-β1 and IL-1β protein and mRNA were significantly lower than the model group′s averages.Conclusions:Extracorporeal shockwave therapy can reduce the expression of TGF-β1 and IL-1β in the cartilage of an arthritic knee, at least in rabbits. Its therapeutic effect is positively correlated with the density of the energy flow, suggesting that shock waves may reduce the expression of inflammatory factor IL-1β by regulating the expression of TGF-β1. They should be applied in the prevention and treatment of osteoarthritis.

ObjectiveTo investigate the effect of Zishen Huoxue prescription on promoting neurogenesis in hippocampal CA1 region of vascular dementia （VD） rats by regulating mitophagy. MethodThe 2-VO method was used to establish the VD rat model and 60 SD rats were randomly divided into sham operation group， model group， donepezil hydrochloride group， and Zishen Huoxue prescription low-dose（8.9 g·kg^-1）， medium-dose（17.8 g·kg^-1） and high-dose（35.6 g·kg^-1） groups. Morris water maze test was performed to detect the escape latency and the number of crossing platform in each group. The expression of phosphatase and tensin homology-induced kinase 1 （PINK1） and Parkinson protein （Parkin） mRNA in hippocampal CA1 region was detected by Real-time fluorescent quantitative polymerase chain reaction（Real-time PCR）. Western blot was used to determine the expression of mitochondrial autophagy signaling pathway-related proteins Parkin， prohibitin 2 （PHB2）， mitofusin 2 （Mfn2） and dynamin-related protein 1 （Drp1） in hippocampal CA1 region. The neurogenesis in hippocampal CA1 region was tested by Brdu method. ResultCompared with the conditions in the sham operation group， the learning and spatial memory abilities of the model group were decreased （P<0.05）， with damaged mitochondrial structure and autolysosome formation in the hippocampal CA1 region. The expressions of Parkin， Pink1 mRNA and Parkin， PHB2， and Drp1 proteins were up-regulated （P<0.05）， while the expression of Mfn2 protein and the neuronal regeneration in hippocampal CA1 region were reduced （P<0.05， P<0.05）. Compared with the conditions in the model group， Zishen Huoxue prescription enhanced the learning and spatial memory abilities of VD rats （P<0.05）， increased the number of autophagosomes in hippocampal CA1 region and improved the mitochondrial structure. The expression of Parkin， Pink1 mRNA and Parkin， PHB2， and Drp1 proteins in hippocampal CA1 region was up-regulated （P<0.05， P<0.01）while the expression of Mfn2 protein was down-regulated（P<0.05， P<0.01）. The number of new neurons in hippocampal CA1 region was also increased（P<0.05， P<0.01）. ConclusionThe promoting effect of Zishen Huoxue prescription on the neurogenesis in hippocampal CA1 region of VD rats was related to the mitophagy mediated by Pink1/Parkin signaling pathway.

Objective:To evaluate the accuracy of two-dimensional ultrasound(2D-US), three-dimensional volume ultrasound (3D-US) and ultrasound strain elastography (USE) in the measurement of preoperative tumor size of breast cancer and its influencing factors.Methods:A total of 101 patients with breast cancer in Nanfang Hospital of Southern Medical University from April to November 2016 were recruited in this study. The maximum diameter of the lesion was examined by 2D-US 3D-US and USE before core needle biopsy or surgery biopsy. The Bland-Altman analysis and intraclass correlation coefficient (ICC) were used to analyze the consistency between the ultrasonic technique measurements and the pathological measurements of postoperative lesion. Chi-square test or Fisher exact test was used to analyze whether the accuracy of three imaging techniques was affected by different clinical pathologic factors and imaging characteristics.Results:3D-US showed better agreement with histology than 2D-US and USE, with a higher ICC (ICC 3D-US=0.90>ICC 2D-US=0.81>ICC SUE=0.78) and low variation. In 3D-US, the accuracy rate of the age >40 years old group was higher than ≤40 years old group. In 2D-US, the measurement accuracy of invasive ductal carcinoma (IDC) without intraductal carcinoma in situ (DCIS) group was higher than DCIS with DCIS group, non-microcalcifications group was more accurate than microcalcifications group. The long diameter of lesion ≤2 cm group was more accurate than >2 cm group, IDC group was more accurate than invasive lobular carcinoma(ILC) group. In USE, the measurement accuracy of IDC without DCIS group was higher than DCIS with DCIS group, non-microcalcifications group was more accurate than microcalcifications group. All the differences mentioned above were statistically significant(all P<0.05). Conclusions:For accurate measurement of the size of breast cancer lesions, 3D-US is the best, which is least affected by clinicopathological factors and imaging features, followed by 2D-US and USE. This has certain significance for clinically determining the extent of breast cancer lesions.

Objective:To investigate the prevalence of Escherchia albertii in Shanxi province. Methods:The chicken intestines were enriched in EC broth. The eae gene was detected by PCR, and the eae-positive EC enrichments were inoculated in MacConkey agar plate. The eae-positive lactose non-fermenting isolates were presumed as Escherchia albertii, and then analyzed by triplex-PCR, 16S rDNA sequencing and MLST. Results:Two suspected Escherchia albertiiwere isolated from 250 samples of chicken intestines. It was identified as Escherchia albertii by phenotypic, specific genes,16S rDNA sequencing, and MLST analyses . The cytolethal distending toxin B ( cdtB) showed positive by PCR,and they were clusted to Ⅱ/Ⅲ/Ⅴ group by sequencing. Conclusion:This study showed that the Escherchia albertii was existed in Shanxi province, China.