Objective To explore the effect and mechanism of Cyclosporin A (CsA) and cholic acid on reducing the human liver cell damage induced by α-amanitin (AMA).Methods According to the previous research results,the minimum hepatocellular survival concentration against αt-AMA (1.4 g/L),the experiment was conducted in 5 groups:control group,damage group,glycochenodeoxycholic acid group,CsA group,and CsA combined with cholic acid group (CsA+ taurocholic acid,CsA+ chenodeoxycholic acid,CsA+ glycocholic acid,CsA+ glycochenodeoxycholic acid,and CsA+ taurochenodeoxycholic acid).For each group,there were 3 time points for observation (24 h,48 h and 72 h after attacking),CsA and CsA+ glycochenodeoxycholic acid was used to protect hepatocytes,respectively,and morphological changes in cells were observed with inverted phase contrast microscope,and the live cells were counted by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) method,and aspertate aminot ransferase (AST) and alanine aminotransferase (ALT) in the culture supernatant were detected by biochemical method.Results Compared with the control group,hepatocellular growth in the injury group was obviously suppressed,with progressive cellular apoptosis and significantly decreased absorbance value of MTIT (0.345 ± 0.021);the activity of AST and ALT increased gradually,reaching the highest after 72 h [(98.4 ± 6.7) U/L and (116.2 ± 9.5) U/L,respectively].Compared with injury group,broken organelles decreased significantly and absorbance value elevated in glycochenodeoxycholic acid group,CsA group and CsA combined with cholic acid group,and at each time point,the highest absorbance value in the CsA+ taurochenodeoxycholic acid group [the highest was (0.656 ± 0.014),P ＜ 0.05];at the same time,the activity of AST and ALT didn't increase obviously,at each time point,the lowest in CsA+ taurochenodeoxycholic acid group [the lowest was (22.3 ± 6.2) U/L and (20.2 ± 5.4) U/L,P ＜ 0.05,respectively].Conclusions CsA,as well as cholic acid,can protect human liver cells from the attack of α-AMA.The mechanism may be CsA,as an organic anion transfer peptide in humans (OATP1B1 and OATP1B3) suppressant,inhibits the absorption of α-AMA.The joint application of CsA and taurochenodeoxycholic acid is superior to the single OATP substrate or inhibitor.

In recent years, cancer stem cells have become a hotspot for global researchers. Cancer stem cell theory deems that cells with self-renewal and differentiation potential play a key role in tumor resistance and relapse. These cells are named cancer stem cells. At present, the sorting methods include the side population cell sorting technique, screening techniques based on cell surface special markers, tumor sphere cultures, label retaining cell, ALDEFLUOR assays and so on. Side population cells are a small part of cells with the capacity of efflux DNA fluorescent dye Hoechst 33342 and present a low staining intensity in flow cytometry plot. Side population cells are rich in cancer stem cells, and its sorting method has been considered simple and effective in cancer stem cell research.
Flow Cytometry ; Humans ; Neoplasms ; Neoplastic Stem Cells ; Side-Population Cells

Objective To investigate the peri-nasopharyngeal invasion patterns of nasopharyngeal carcinoma (NPC) on MRI and its relationship with tumor staging. Methods One thousand five hundred and seventy-three patients with newly diagnosed NPC which were histo-pathologically proved were retrospectively studied. The MRI manifestations and invasion patterns of the NPCs were elevated according to the 2008 Tstaging system of NPC. Z test was used to analyze the rate of adjacent structures invasion in NPCs. Results The structures invaded by NPCs included pharyngobasilar fascia in 1299 cases (82. 58% ); parapharyngeal space, 1090 ( 69. 29% ); nasal cavities, 304 ( 19. 33% ); oropharynx, 49 ( 3. 12% ); carotid space,514(32. 68% ); medial pterygoid muscle, 661 (42. 02% ); lateral pterygoid muscle, 210( 13. 35% ); skull base bones, 943(59. 95% ); cranial nerves, 630(40. 05% ) and paranasal sinuses, 242 ( 15.38% ). The T-stage distribution was T1, 242 cases ( 15.38% ); T2, 288 ( 18. 31% ); T3, 410 (26. 06% ) and T4,633 (40. 24% ). Among the cases with nasal cavities invasion, 90. 46% (275/304)showed the involvement of the structures seen in T3 or T4 stage, which was found in all cases with oropharynx invasion. In addition,69. 14% (457/661) of cases with medial pterygoid muscle invasion and 92. 15% (223/242) of cases with paranasal sinuses invasion showed the involvement of structures seen in T4 stage. As for the invasion patterns of NPC, the lateral invasion of pharyngobasilar fascia was more frequent than upward invasion of skull base (Z = 14. 025, P ＜ 0. 01 ) and downward invasion of oropharynx ( Z = 45.032, P ＜ 0. 01 ), and the downward invasion of oropharynx was less frequent than upward invasion of skull base ( Z = 34. 301, P ＜ 0. 01 ) and forward invasion of nasal cavities ( Z = 14. 404, P ＜ 0. 01 ). Conclusion NPC has a predilection of lateral invasion rather than upward and downward invasion, and its upward and forward invasion are more common than downward invasion.

Objective To investigate the application value of deep learning optical coherence tomography ( OCT) on wet age-related macular degeneration ( wAMD) assisted diagnosis. Methods Weakly supervised deep learning algorithms was applied on the premise that only disease or not can be provided as a marker. The OCT image automatically assisted in the diagnosis of diseased areas of wAMD,and thermograms were applied to provide a basis for doctors to detect disease areas. Based on the deep learning of weak supervision,a new network algorithm structure was proposed for detecting disease area in ophthalmic OCT images. At the same time, thermograms were adopted to improve the accuracy of the lesion map,which is the location of the lesion area. This study followed the Declaration of Helsinki. This study protocol was approved by Ethic Committee of Ningbo Eye Hospital ( No. 2018-YJ05 ) . Written informed consent was obtained from each subject before entering study cohort. Results Resnet-based deep learning algorithm gave a diagnostic accuracy rate of 94. 9％ for the disease, which was much higher than that of AlexNet 85. 3％,VGG 88. 7％, and Google-Net 89. 2％. The thermograms with different colors provided a more convenient auxiliary diagnosis basis for doctors. Conclusions Compared with the original classification network,which needs disease area markers as empirical knowledge, deep learning algorithm model not only provides better results in the classification of retinal diseases,but also marks potential disease areas. The lesion area provides a basis for judging the area of the lesion for the diagnosis of wAMD.

Objective:To investigate the protective effect and mechanism of silymarin on bronchiolitis caused by respiratory syncytial virus (RSV) in mice.Methods:A mouse model of bronchiolitis was established by intranasal instillation of RSV. After successful modeling, the mice were randomly divided into a model group, a positive control group (ribavirin, 10 mg/kg), a low-dose silymarin group (25 mg/kg), a medium-dose silymarin group (50 mg/kg), and a high-dose silymarin group (100 mg/kg). In addition, a control group was established, with 12 mice in each group. The pulmonary index and RSV virus load were determined in each group of mice. Hematoxylin-eosin (HE) staining was used to observe the pathological changes in the lungs. The levels of interleukin-4 (IL-4), interferon-γ (IFN-γ), transforming growth factor-β (TGF-β), IL-17, and IL-10 in bronchoalveolar lavage fluid (BALF) were detected by enzyme-linked immunosorbent assay (ELISA). Flow cytometry was used to detect the proportion of helper T cells 17 (Th17) and regulatory T cells (Treg) in peripheral blood. Western blot was used to detect the expression levels of Toll-like receptor 4 (TLR4), myeloid differentiation factor 88 (MyD88), nuclear factor κB subunit p65 (NF-κB p65), and phosphorylated nuclear protein p-NF-κB p65 in lung tissue.Results:Compared with the model group, the pulmonary injury and inflammatory response were significantly improved in the medium-and high-dose silymarin groups. The pulmonary indexes were (1.27±0.17)% and (0.94±0.10)%, respectively, and the RSV virus loads were (2.65±0.19) and (2.13±0.14), respectively, which were significantly lower than those in the model group (all P<0.05). The proportion of Th17 cells in peripheral blood was (4.47±0.19)% and (3.52±0.13)%, respectively, which was significantly lower than that in the model group (all P<0.05), while the proportion of Treg cells in peripheral blood was (0.88±0.08)% and (1.33±0.12)%, respectively, which was significantly higher than that in the model group (all P<0.05). The expression levels of IL-4 and IL-17 in BALF and the protein expression levels of TLR4/NF-κB signaling pathway related proteins in lung tissue were significantly lower than those in the model group (all P<0.05), while the expression levels of IFN-γ, TGF-β, and IL-10 in BALF were significantly higher than those in the model group (all P<0.05). Conclusions:Silymarin can regulate immune function and inhibit inflammatory response, thereby improving airway inflammation in bronchiolitis mice. The mechanism may be related to inhibit activation of TLR4/NF-κB signaling pathway.

Background: Excessive delivery of free fatty acids (FFAs) to the liver promotes steatosis and insulin resistance (IR), with IR defined as reduced glucose uptake, glycogen synthesis and anti-lipolysis stimulated by normal insulin levels. Whether the associations between FFAs and diabetes development differ between patients with and without nonalcoholic fatty liver disease (NAFLD) remains unclear. Methods: Consecutive subjects (2,220 NAFLD subjects and 1,790 non-NAFLD subjects according to ultrasound imaging) were enrolled from the First Affiliated Hospital of Sun Yat-sen University between 2009 and 2019. The homeostasis model assessment of insulin resistance (HOMA-IR) was calculated. Results: There was an approximate J-shaped relationship between FFA levels and HOMA-IR in the NAFLD group. Higher FFA concentration quartiles were associated with higher risks of IR (odds ratio [OR], 9.24; 95% confidence interval [CI], 6.43 to 13.36), prediabetes (OR, 10.48; 95% CI, 5.66 to 19.39), and type 2 diabetes mellitus (T2DM; OR, 19.43; 95% CI, 12.75 to 29.81) in the NAFLD group but not in the non-NAFLD group. The cut-off points for the FFA levels increased in a stepwise manner in discriminating IR, prediabetes and T2DM (573, 697, and 715 μmol/L) in the NAFLD group but not in non-NAFLD individuals. Conclusion A distinct dose-dependent relationship of FFA levels was found with IR, prediabetes and T2DM in NAFLD patients. Screening serum FFA levels in NAFLD patients would be valuable in preventing diabetes development.

Background: Excessive delivery of free fatty acids (FFAs) to the liver promotes steatosis and insulin resistance (IR), with IR defined as reduced glucose uptake, glycogen synthesis and anti-lipolysis stimulated by normal insulin levels. Whether the associations between FFAs and diabetes development differ between patients with and without nonalcoholic fatty liver disease (NAFLD) remains unclear. Methods: Consecutive subjects (2,220 NAFLD subjects and 1,790 non-NAFLD subjects according to ultrasound imaging) were enrolled from the First Affiliated Hospital of Sun Yat-sen University between 2009 and 2019. The homeostasis model assessment of insulin resistance (HOMA-IR) was calculated. Results: There was an approximate J-shaped relationship between FFA levels and HOMA-IR in the NAFLD group. Higher FFA concentration quartiles were associated with higher risks of IR (odds ratio [OR], 9.24; 95% confidence interval [CI], 6.43 to 13.36), prediabetes (OR, 10.48; 95% CI, 5.66 to 19.39), and type 2 diabetes mellitus (T2DM; OR, 19.43; 95% CI, 12.75 to 29.81) in the NAFLD group but not in the non-NAFLD group. The cut-off points for the FFA levels increased in a stepwise manner in discriminating IR, prediabetes and T2DM (573, 697, and 715 μmol/L) in the NAFLD group but not in non-NAFLD individuals. Conclusion A distinct dose-dependent relationship of FFA levels was found with IR, prediabetes and T2DM in NAFLD patients. Screening serum FFA levels in NAFLD patients would be valuable in preventing diabetes development.

Objective To examine the association between social isolation and cognitive function among the elderly living in the communities of Daqing city.Methods A total of 981 community residents aged 60 years or over,were surveyed with a questionnaire.Both Lubben Social Network Scale-6 (LSNS-6) and Montreal Congnitive Assessment (MoCA) Changsha Versions were used to respectively screen the status of social isolation and cognitive function,on these elderly.Results The average age was 71 years old for the 981 study participants.10.60％ (104/981) of the participants were assessed as having the status of social isolation,9.48％ (93/981) as having marginal family ties and 13.97％ (137/981) as having marginal friendship ties.Results from the multivariate linear regression analysis revealed that participants with higher scores of LSNS-6 presenting better cognitive function score,with a partial regression coefficient as 0.10 (P＜0.01).The MoCA scores in participants with social isolation (20.38 ± 5.54) were significantly lower than the ones without social isolation (22.10 ± 5.01) and the difference was statistically significant (P＜0.01).Social isolation was significantly related to the domain scores on visuo-spatial constructional executive functions (P=0.02),naming (P=0.03),language (P=0.01) and delayed memory functions (P＜0.01),but not with other domains as concentration (P=0.33),orientation (P=0.27) or abstraction (P=0.49).Conclusion The findings suggested that social isolation was mainly caused by the lack of friendship ties and associated with cognitive function and among the elderly in Daqing city,Heilongjiang province.

BACKGROUND:Cellular senescence is closely related to the development and progression of osteoarthritis,but the specific targets and regulatory mechanisms are not yet clear. OBJECTIVE:To mine key genes in cellular senescence-mediated osteoarthritis by integrating bioinformatics and machine learning approaches and validate them via experiments to explore the role of cellular senescence in osteoarthritis. METHODS:The osteoarthritis gene expression profiles obtained from the GEO database were intersected with cellular senescence-related genes obtained from the CellAge database and the expression of the intersected genes was extracted for differential analysis,followed by GO and KEGG analysis of the differential genes.The key osteoarthritis cellular senescence genes were then screened by protein-protein interaction network analysis and machine learning,and in vitro cellular experiments were performed.Finally,the expression of the key genes was detected by qPCR. RESULTS AND CONCLUSION:A total of 31 osteoarthritis cell senescence differential genes were identified.GO analysis showed that these genes were mainly involved in the biological processes,such as regulation of leukocyte differentiation,monocyte differentiation,regulation of T cell differentiation and exerted roles in DNA transcription factor binding,histone deacetylase binding,chromatin DNA binding,and chemokine binding.KEGG analysis showed that osteoarthritis cell senescence differential genes were mainly activated in the JAK/STAT signaling pathway,PI3K/Akt signaling pathway and FoxO signaling pathway.MYC,a key gene for osteoarthritis cellular senescence,was identified by protein-protein interaction network topology analysis and machine learning methods.The results of the in vitro cellular assay showed that the mRNA expression of MYC was significantly lower in the experimental group(osteoarthritis group)than the control group(normal group)(P＜0.05).To conclude,MYC can be a key gene in the senescence of osteoarthritic cells and may be a new target in the prevention and treatment of osteoarthritis by mediating immune response,inflammatory response and transcriptional regulation.

Objective The aim of this study was to evaluate the efficacy and side effects of sacubitril/valsartan in the treatment of patients with chronic kidney disease(CKD)at stage 5 with resistant hypertension,and to explore the cardiovascular benefits and security of medical in the patients.Methods Patients with CKD5 resistant hypertension diagnosed and treated in the First Affiliated Hospital of Guangxi Medical University from September 2020 to March 2022 were selected and divided into the observation group(treated with routine treatment of kidney disease at end-stage and sacubitril/valsartan)and control group(include droutine treatment of renal disease at end-stage and ACEI or ARB drugs)according to treatment strategy.The patients in both two groups were treated with adequate dialysis treatment and conventional drug treatment of renal disease at end-stage.The patients were followed up for at least 3 months,the clinical efficacy of three months after treated with sacubitril/valsartan was observed,and the efficacy indicators and security indicators and adverse cardiovascular events were observed,the occurrence of adverse effects during the period of drug use were compared with the control group.Results A total of 110 patients were included in this study and there were 55 cases in each group.There were no significant differences in gender,age,age of dialysis,etiology,dialysis mode and blood pressure between the two groups(P>0.05).The Systolic blood pressure(SBP),diastolic blood pressure(DBP),b-type urinary natriuretic peptide precursor(Pro-BNP)and cardiac function grade in the observation group after treatment was significantly decreased compared with before treatment.The left ventricular ejection fraction(LVEF)and the ratio of LVEF<50%in the observation group was significantly reduced after treatment(P<0.05).SBP,DBP and Pro-BNP decreased 3 months after treatment compared with the baseline before treatment,and improved significantly in the first month after treatment(P<0.05).The decrease of DBP and BNP before and after treatment was significantly different between the two groups,and the decrease of DBP and BNP was more significant in the observation group(P<0.05).The difference of LVEF and left ventricular end diastolic diameter(LVEDD)between the two groups before and after treatment was statistically significant,and the improvement was more obvious in the observation group(P<0.05).There were no significant differences in the safety indicators of serum potassium,estimated glomerular filtration rate(eGFR)and liver function between two groups before and after treatment(P>0.05).In terms of adverse reactions,only 1 case in the control group developed hyperkalemia within 3 months of follow-up,and no hypotension or other adverse reactions occurred in the two groups.Conclusions The treatment of patients with CKD stage 5 hypertension with sacubitril/valsartan has obvious cardiovascular benefits.Sacubitril/Valsartan has efficacy in lowering blood pressure,improving cardiac function and reducing volume load,with less adverse events and higher safety than control group.