BACKGROUND/OBJECTIVES: Previous research has shown maternal betaine supplementation alleviates fetal-derived hepatic steatosis. Therefore, this study examined the anti-inflammatory effect of maternal betaine intake in offspring mice and its mechanism.MATERIALS/METHODS: Female C57BL/6J mice and their offspring were randomly divided into 3 groups according to the treatment received during gestation and lactation: control diet (CD), fatty liver disease (FLD), and fatty liver disease + 1% betaine (FLD-BET). The FLD group was given a high-fat diet and streptozotocin (HFD + STZ), and the FLD-BET group was treated with HFD + STZ + 1% betaine. After weaning, the offspring mice were given a normal diet for 5 weeks and then dissected to measure the relevant indexes. RESULTS: Compared to the CD group, the offspring mice in the FLD group revealed obvious hepatic steatosis and increased serum levels of alanine aminotransferase, interleukin (IL)-6, and tumor necrosis factor (TNF)-α; maternal betaine supplementation reversed these changes. The hepatic mRNA expression levels of IL-6, IL-18, and Caspase-1 were significantly higher in the FLD group than in the CD group. Maternal betaine supplementation reduced the expression of IL-1β, IL-6, IL-18, and apoptosis-associated speck-like protein containing C-terminal caspase recruitment domain (ASC). Maternal betaine supplementation also reversed the increasing protein expressions of nitric oxide dioxygenase-like receptor family pyrin domain containing 3 (NLRP3), ASC, Caspase-1, IL-1β, and IL-18 in offspring mice exposed to HFD + STZ. Maternal betaine supplementation decreased the homocysteine (Hcy) and s-adenosine homocysteine (SAH) levels significantly in the livers. Furthermore, the hepatic Hcy concentrations showed significant inverse relationships with the mRNA expression of TNF-α, NLRP3, ASC, and IL-18. The hepatic SAH concentration was inversely associated with the IL-1β mRNA expression. CONCLUSIONS The lipotropic and anti-inflammatory effect of maternal betaine supplementation may be associated with the inhibition of NLRP3 inflammasome in the livers of the offspring mice.

Objective:To preliminarily analyze the expression of angiotensin-converting enzyme 2 (ACE2) and to investigate its potential regulatory mechanism in chronic rhinosinusitis with nasal polyps (CRSwNP).Methods:Patients underwent nasal endoscopic surgery in the Third Affiliated Hospital of Sun Yat-sen University from February 2020 to May 2021 were selected, including 17 males and 6 females, aging from 23 to 66 years old. Expression of ACE2 was evaluated via immunohistochemical staining in controls with non-chronic rhinosinusitis, non-eosinophilic CRSwNP (non-ECRSwNP), and eosinophilic CRSwNP (ECRSwNP) tissue, respectively. Correlations between ACE2 and the indicated Th1/Th2-related cytokines (IFN-γ, IL-4, IL-5, IL-13, IL-25, IL-33, TSLP and periostin) were analyzed based on GSE72713 dataset. Protein-protein interaction (PPI) network was constructed via string database, immune infiltration of GSE72713 dataset was evaluated using cibersort algorithm. ACE2 was comprehensively analyzed by microRNA regulatory network, gene set enrichment analysis (GSEA) and pharmacological analysis. Statistical analysis was performed using GraphPad 7.0 and SPSS 20.0 software.Results:ACE2 was up-regulated in non-ECRSwNP compared with ECRSwNP. Microarray analysis showed that ACE2 was positively correlated with IFN-γ while inversely correlated with IL-5, IL-13 and periostin significantly. Analysis of immune infiltration suggested that ACE2 expression correlated positively with the number of M1 macrophage while negatively with M2 macrophage. GSEA demonstrated that interferon-related signaling pathways were up-regulated in non-ECRSwNP, and miRNA-200B/miRNA-200C/miRNA-429 pathways targeting ACE2 were enriched in ECRSwNP. Results of pharmacological analysis indicated that ampicillin was able to promote the expression of ACE2 whereas acetaminophen could down regulated the expression of ACE2.Conclusion:Expression pattern of ACE2 is varied in non-ECRSwNP and ECRSwNP, which may be related to the different infiltration of indicated cytokines and different regulatory pathways of miRNA.

AIM: To investigate the effect of nobiletin (Nob) on cardiomyocyte hypertrophy induced by high glucose and its mechanism. METHODS: Neonatal rat cardiomyocytes (NRCMS) were stimulated with high glucose (HG) to establish cardiomyocyte hypertrophy and nobiletin was given. Cell viability was measured by MTT assay. C-myc and Nppa mRNA levels were detected by qRT-PCR. Cellular surface area was detected by immunofluorescence, and Nrf2 and HO-1 protein expressions were detected by Western blot. RESULTS: After stimulation with 33.3 mmol/L HG for 48 h, the survival rate of NRCMS was significantly decreased, C-myc, Nppa mRNA levels and cellular surface area were significantly increased, Nrf2 and HO-1 protein expression were significantly decreased (P<0.05). After Nob treatment, compared with HG group, cellular surface area, Nrf2 and HO-1 protein expression were significantly increased, C-myc and Nppa mRNA levels were significantly decreased. The above indexes were reversed by using Nrf2 inhibitor. CONCLUSION: Nob inhibits cardiomyocyte hypertrophy induced by high glucose, and its mechanism may be related to the activation of Nrf2/HO-1 signaling pathway.

Objective:To investigate the expression and cellular provenance of interleukin 17A (IL-17A) in non-eosinophilic chronic rhinosinusitis with nasal polyps (nECRSwNP), and to analyze the possible reasons for its different expression.Methods:Samples were collected from 14 patients with eosinophilic chronic rhinosinusitis with nasal polyps (ECRSwNP) and 28 patients with nECRSwNP, who underwent functional endoscopic sinus surgery in the Third Affiliated Hospital of Sun Yat-sen University from January 2017 to May 2018, including 33 males and 9 females, with the age ranging from 18 to 65 years old. Enzyme linked immune sorbent assay (ELISA) and flow cytometry were used to investigate the expression and cellular origins of IL-17A in the nasal tissue of ECRSwNP and nECRSwNP groups. Then the difference of quantity and differentiation ability of the major cells producing IL-17A between ECRSwNP and nECRSwNP groups were analyzed by flow cytometry. Finally, the expressions of IL-6, transforming growth factor-β(TGF-β), and IL-23, which were considered as the important factors in promoting Th17/Tc17 differentiation in CRSwNP and their correlation with IL-17A, were analyzed by ELISA. Statistical analysis was performed using IBM SPSS 20.Results:The IL-17A protein levels and IL-17A +lymphocyte percentages were higher in nECRSwNP group compared with that of the ECRSwNP group (158.56 (167.76) pg/ml ( M( QR)) vs. 9.42 (11.33) pg/ml, 10.21%±1.54% ( ± s) vs. 3.93%±0.80%, Z=2.95, t=3.62, all P<0.01). Tc17 cells (CD8 +T cells producing IL-17A) and Th17 cells (CD4 +T cells producing IL-17A) were major IL-17A producers in both ECRSwNP and nECRSwNP group. Further analysis revealed that there was no significant difference in quantity of CD8 +and CD4 +T cells between ECRSwNP and nECRSwNP group, but the differentiation ability about CD8 +and CD4 +T cells differentiating into Tc17 and Th17 in nECRSwNP group was stronger than that in ECRSwNP. The high expressions of IL-6 and TGF-β, which were considered as the important factors in promoting Th17/Tc17 differentiation were also found in nECRSwNP group compared with ECRSwNP (56.07 (234.25) pg/ml vs. 8.27 (12.51) pg/ml, (5.44±0.34) pg/ml vs. (4.17±0.22) pg/ml, Z=2.426, t=2.29, all P<0.05). But the difference in expression of IL-23 was not significant difference between the two groups. Moreover, the expression of IL-17A showed significantly positive correlation with IL-6 ( r=0.615, P=0.009). No positive correlation between IL-17A and TGF-β or IL-23 was observed. Conclusions:The expression of IL-17A in nasal mucosa of nECRSwNP patients is significantly higher than that of ECRSwNP, which is due to the increase of expression and differentiation of Tc17/Th17 cells. IL-17A shows positive correlation with IL-6 in CRSwNP, which is the important factor in promoting Th17/Tc17 differentiation.

Objective To investigate the characteristics and significance of sural sparing pattern and radial sparing pattern in children immune-mediated acute or chronic inflammatory peripheral neuropathy.Methods Forty children with Guillain-Barre syndrome (GBS) and chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) aged<14 years, admitted to our hospital from September 2014 to August 2019, were chosen as immune-inflammatory group; 15 children (<14 years old) with multiple peripheral neuropathy of other etiologies from the same source were classified as other etiologies group. The characteristics of sensory nerve damage measured by nerve conduction velocity in the two groups were compared. The existences of sural sparing (SS) pattern and radial sparing (RS) pattern, and sensory ratio (SR)>1 were conformed. SR=(sural nerve sensory nerve action potentials+radial nerve sensory nerve action potentials)/(median nerve sensory nerve action potentials+ulnar nerve sensory nerve action potentials).Results (1) In 40 patients from immune-inflammatory group, 38 were GBS and two were with CIDP; Among the 38 with GBS, 34 were with acute inflammatory demyelinatingpolyneuropathies (AIDP), 3 were with acute motor axonal neuropathy (AMAN), and one was with acute motor sensory axonal neuropathy. Among the 15 patients from the other etiologies group, 9 were with charcot-marie-tooth (CMT) type 1, one was with CMT type 2, one was with CMT intermediate type, and two were with hereditary neuropathy with liability to pressure palsies (HNPP), and two were with mitochondrial disease involves polyneuropathy. (2) As compared with patients from other etiologies group, patients from immune-inflammatory group had significantly higher positive rates of SS (72.5％vs. 6.7％) , RS(42.5％vs.6.7％) and SR>1 (75.0％vs.13.3％). In the diagnoses of childhood acute or chronic inflammatory polyneuropathy, the sensitivity of SS pattern was 0.73, specificity was 0.93, and positive likelihood ratio was 10.88. SR>1 had moderate specificity (0.87) and low sensitivity (0.41).Conclusion The sensory sparing patterns, especially SS pattern, have valuable clinical significance in the diagnoses of acute or chronic inflammatory peripheral neuropathy in children.

Objective:To investigate the pro-apoptotic effect of berberine (Ber) on human lung adenocarcinoma PC-9 cell line, and to detect the role of c-Jun N-terminal kinase (JNK)/forkhead box protein O3 (FOXO3) signaling in this process. Methods:The PC-9 cells were randomly divided into the control group and the Ber group, which was treated with 30 and 60μM Ber. The survival rate, apoptot-ic rate, ROS generation, caspase-3 activity, and mitochondrial membrane potential of cells were detected. Western blot was per-formed to detect the expression of JNK/FOXO3 signaling and apoptosis-related proteins. A JNK-specific activation inhibitor, SP600125, was used to block the phosphorylation of FOXO3 in PC-9 cells, and then treated with Ber (30 and 60μM) for further detection after 24 h. Results:Ber treatment resulted in an obvious reduction in cell viability, promotion of cell apoptosis, downregulation of mitochondri-al membrane potential, and an increase of ROS and caspase-3 in a dose-dependent manner. Western blot analysis demonstrated that Ber treatment resulted in a significant upregulation of p-JNK, FOXO3, and Bax expression, and a downregulation of p-FOXO3 and Bcl2 levels. Moreover, the inhibition of JNK activation by SP600125 antagonized the anti-FOXO3 phosphorylation role and the pro-apoptotic role of Ber on PC-9 cells. Conclusion:Ber treatment effectively inhibits the viability of PC-9 cells and enhances apoptosis and oxidative stress injury, which may be related to the upregulation of p-JNK and FOXO3 and the downregulation of p-FOXO3.

Genetically modified tool animal models are the animal models,which are generated by modifying a defined gene and can be used as a tool to help realize other objective.Genetically modified large animals have wide applications in agriculture and biomedicine.Tool animal models play important role in biological research and development of new drugs in biomedicine,especially,have made tremendous contribution in revealing gene function and pathway of signal transduction.Pigs are not only an economically important agriculture animals,but also an ideal animal model in biomedicine due to its close similarity to human in physiology,as well as organ structure and size.Thanks to the breakthrough of newly emerged gene editing technology,striking progress has made in establishment of genetically modified tool pig models which include the ones used for monitoring pluripotency of cells,tracing cell lineages,replacing genes mediated by Cre recombinase,mimicking immunodeficiency,as well as gene editing in vivo.These tool models have been widely applied in biological research.Here,we will review the progress in generation of genetically modified tool pig models and their applications.

Objective: To research the best processing method for ginger pinellia by orthogonal tests.Methods: The orthogonal tests included the soaking time, boiling water and cooking time as the influencing factors, an HPLC method was used for the determination of 4 nucleosides (uridine, guanosine, adenosine, inosine), and the alum limit and extract content were also studied.The results were evaluated by multi index comprehensive weighted score to optimize the processing technology of ginger pinellia.Results: The best processing technology of ginger pinellia was as follows: soaked for 60 hours, the proportion of boiling water and pinellia tuber was 15:1, and boiled for about 5 h.Conclusion: The optimum processing technology of ginger pinellia is reasonable, reliable and reproducible, which can be used as the reference for the processing standardization of Chinese crude drugs.

Objective To establish a method of quick three-dimensional (3D) reconstruction of pubic symphysis based on magnetic resonance imaging. Methods The pelvis images of adult male were generated on a 3.0 T scanner using a T1 Gradient Echo FLASH-3D (T1- FL3D) sequence and imported the images into medical image control system. Segmentation of binaryzation threshold was conducted and pelvic soft tissue image was extracted by regional growth, 3D structure model of pubic symphysis was obtained by Boolean operation. The 3D structure model of pubic symphysis was established by the noise reduction of reverse engineering software. And compared with the 3D reconstruction model pubic bone CT scan. Results The morphological characters of the MRI pubic symphysis 3D model, such as the ridges and furrows on the symphysial surface, lower extremity, dorsal margin (beveling), margin (beveling) and pubic tubercle, were highly consistent with the morphological characters of the 3D model established by CT scan. Conclusion MRI scan can be used to reconstruct the 3D structure of pubic symphysis quickly and effectively, and it can provide a safe radiation-free 3D visualization imaging technique for forensic age estimation for the living.