AIM:This study aims to investigate the effects of aerobic exercise on hypothalamic autophagy and central leptin resistance in obese mice,and to explore the potential mechanisms.METHODS:Forty male C57BL/6J mice,aged 7 to 8 weeks,were randomly assigned to 5 groups:normal control(CON)group,high-fat diet(HFD)group,HFD+exercise(HFD+Exe)group,HFD+phosphate-buffered saline(PBS)group,and HFD+rilmenidine(autophagy ago-nist)group,with 8 mice in each group.Additionally,twelve fibronectin type Ⅲ domain-containing protein 5(Fndc5)gene(encoding irisin precursor protein)knockout(Fndc5 KO)mice were randomly allocated to Fndc5 KO+HFD group and Fndc5 KO+HFD+Exe group,with 6 mice in each group.All mice were fed for 28 weeks.The mice in CON group re-ceived a normal diet,while those in the remaining groups were provided with an HFD.The mice in HFD+Exe and Fndc5 KO+HFD+Exe groups engaged in aerobic treadmill exercise while continuing an HFD from weeks 17 to 28.The mice in HFD+PBS group received intraperitoneal injections of PBS as a control,while those in HFD+rilmenidine group received in-traperitoneal injections of rilmenidine(10 mg?kg-1?d-1),4 times a week over a total duration of 12 weeks(weeks 17 to 28).Following the intervention,serum metabolite levels,as well as concentrations of leptin and irisin,were quantified by ELISA.Morphological alterations in the liver and white adipose tissues were evaluated through oil red O staining and he-matoxylin-eosin(HE)staining.Western blot was utilized to assess the hypothalamic protein levels of autophagy markers,autophagy-related protein 7(ATG7),beclin-1,microtubule-associated protein 1 light chain 3(LC3)and p62,and leptin resistance markers,suppressor of cytokine signaling 3(SOCS3)and protein tyrosine phosphatase 1B(PTP1B).RE-SULTS:Observations of mouse phenotypes indicated that HFD feeding significantly increased body weight,blood lipid content and serum leptin level(P<0.05).The results of HE and oil red O staining demonstrated that HFD feeding marked-ly promoted lipid accumulation in the liver and caused ballooning of white adipocytes.Western blot analyses revealed that HFD feeding significantly down-regulated the protein levels of ATG7,beclin-1 and LC3-Ⅱ/LC3-I,but up-regulated the pro-tein level of p62(P<0.05),thus reducing cellular autophagy capacity.Furthermore,HFD feeding elevated the protein levels of leptin resistance markers SOCS3 and PTP1B(P<0.05).Aerobic exercise and autophagy agonist were found to partially reverse these changes,enhancing cellular autophagy capacity and alleviating leptin resistance.However,these effects were diminished after knockout of Fndc5 gene,further substantiating the role of irisin in exercise-mediated en-hancement of cellular autophagy and attenuation of leptin resistance.CONCLUSION:Aerobic exercise alleviates hypo-thalamic autophagy defect and central leptin resistance in obese mice,which may be associated with exercise-induced irisin.

OBJECTIVE To explore the functional substance basis of Jinhong Tablet in the treatment of chronic superficial gastri-tis(CSG).METHODS Three different models were constructed to investigate the anti-inflammatory and antioxidant effects,func-tional material basis of Jinhong Tablet:inflammatory model in human gastric epithelial cells(GES-1)induced by lipopolysaccharide(LPS),LPS-induced inflammatory model in mouse gastric organoids,and ethanol-induced oxidative damage model in GES-1 cells.MTS assay was performed to detect cell proliferation activity;qPCR was applied to measure the relative mRNA expression of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),interleukin-6(IL-6),and interleukin-8(IL-8)in cells and gastric organoids;and the levels of superoxide dismutase(SOD),malondialdehyde(MDA),and reactive oxygen species(ROS)in cells were detected.RESULTS Jinhong Tablet and 10 functional components significantly reduced the relative expression of inflammation-relat-ed genes TNF-α,IL-1β,IL-6,and IL-8 in LPS-induced GES-1 cells and gastric organoids,suggesting that these 10 components are the functional substance basis for the anti-inflammatory effects of Jin Hong Tablet.Jin Hong Tablet and 11 functional components markedly decreased the levels of MDA and ROS and increased the activity of SOD,indicating that these 11 components were the func-tional substance basis of the antioxidant effects of Jinhong Tablet.CONCLUSION Through in vitro cell and gastric organoid experi-ments,it has been preliminarily determined that allocryptopine,corydaline,dehydrocorydaline,palmatine hydrochloride,chlorogenic acid,costunolide,rutin,quercitrin,dehydrocostus lactone,tetrahydrocoptisine,isochlorogenic acid B,toosendanin,protopine,and quercetin are the functional material basis of Jinhong Tablet in treating CSG,accumulating scientific evidence for the enhancement of the quality standards of Jinhong Tablet.

OBJECTIVE To explore the functional substance basis of Jinhong Tablet in the treatment of chronic superficial gastri-tis(CSG).METHODS Three different models were constructed to investigate the anti-inflammatory and antioxidant effects,func-tional material basis of Jinhong Tablet:inflammatory model in human gastric epithelial cells(GES-1)induced by lipopolysaccharide(LPS),LPS-induced inflammatory model in mouse gastric organoids,and ethanol-induced oxidative damage model in GES-1 cells.MTS assay was performed to detect cell proliferation activity;qPCR was applied to measure the relative mRNA expression of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),interleukin-6(IL-6),and interleukin-8(IL-8)in cells and gastric organoids;and the levels of superoxide dismutase(SOD),malondialdehyde(MDA),and reactive oxygen species(ROS)in cells were detected.RESULTS Jinhong Tablet and 10 functional components significantly reduced the relative expression of inflammation-relat-ed genes TNF-α,IL-1β,IL-6,and IL-8 in LPS-induced GES-1 cells and gastric organoids,suggesting that these 10 components are the functional substance basis for the anti-inflammatory effects of Jin Hong Tablet.Jin Hong Tablet and 11 functional components markedly decreased the levels of MDA and ROS and increased the activity of SOD,indicating that these 11 components were the func-tional substance basis of the antioxidant effects of Jinhong Tablet.CONCLUSION Through in vitro cell and gastric organoid experi-ments,it has been preliminarily determined that allocryptopine,corydaline,dehydrocorydaline,palmatine hydrochloride,chlorogenic acid,costunolide,rutin,quercitrin,dehydrocostus lactone,tetrahydrocoptisine,isochlorogenic acid B,toosendanin,protopine,and quercetin are the functional material basis of Jinhong Tablet in treating CSG,accumulating scientific evidence for the enhancement of the quality standards of Jinhong Tablet.

AIM:This study aims to investigate the effects of aerobic exercise on hypothalamic autophagy and central leptin resistance in obese mice,and to explore the potential mechanisms.METHODS:Forty male C57BL/6J mice,aged 7 to 8 weeks,were randomly assigned to 5 groups:normal control(CON)group,high-fat diet(HFD)group,HFD+exercise(HFD+Exe)group,HFD+phosphate-buffered saline(PBS)group,and HFD+rilmenidine(autophagy ago-nist)group,with 8 mice in each group.Additionally,twelve fibronectin type Ⅲ domain-containing protein 5(Fndc5)gene(encoding irisin precursor protein)knockout(Fndc5 KO)mice were randomly allocated to Fndc5 KO+HFD group and Fndc5 KO+HFD+Exe group,with 6 mice in each group.All mice were fed for 28 weeks.The mice in CON group re-ceived a normal diet,while those in the remaining groups were provided with an HFD.The mice in HFD+Exe and Fndc5 KO+HFD+Exe groups engaged in aerobic treadmill exercise while continuing an HFD from weeks 17 to 28.The mice in HFD+PBS group received intraperitoneal injections of PBS as a control,while those in HFD+rilmenidine group received in-traperitoneal injections of rilmenidine(10 mg?kg-1?d-1),4 times a week over a total duration of 12 weeks(weeks 17 to 28).Following the intervention,serum metabolite levels,as well as concentrations of leptin and irisin,were quantified by ELISA.Morphological alterations in the liver and white adipose tissues were evaluated through oil red O staining and he-matoxylin-eosin(HE)staining.Western blot was utilized to assess the hypothalamic protein levels of autophagy markers,autophagy-related protein 7(ATG7),beclin-1,microtubule-associated protein 1 light chain 3(LC3)and p62,and leptin resistance markers,suppressor of cytokine signaling 3(SOCS3)and protein tyrosine phosphatase 1B(PTP1B).RE-SULTS:Observations of mouse phenotypes indicated that HFD feeding significantly increased body weight,blood lipid content and serum leptin level(P<0.05).The results of HE and oil red O staining demonstrated that HFD feeding marked-ly promoted lipid accumulation in the liver and caused ballooning of white adipocytes.Western blot analyses revealed that HFD feeding significantly down-regulated the protein levels of ATG7,beclin-1 and LC3-Ⅱ/LC3-I,but up-regulated the pro-tein level of p62(P<0.05),thus reducing cellular autophagy capacity.Furthermore,HFD feeding elevated the protein levels of leptin resistance markers SOCS3 and PTP1B(P<0.05).Aerobic exercise and autophagy agonist were found to partially reverse these changes,enhancing cellular autophagy capacity and alleviating leptin resistance.However,these effects were diminished after knockout of Fndc5 gene,further substantiating the role of irisin in exercise-mediated en-hancement of cellular autophagy and attenuation of leptin resistance.CONCLUSION:Aerobic exercise alleviates hypo-thalamic autophagy defect and central leptin resistance in obese mice,which may be associated with exercise-induced irisin.

OBJECTIVE: To compare the clinical therapeutic effect between electroacupuncture at "four points of sacral region" and transurethral Erbium laser in treatment of moderate to severe stress urinary incontinence after radical prostatectomy. METHODS: A total of 68 patients of moderate to severe stress urinary incontinence after radical prostatectomy were divided into an electroacupuncture group (34 cases) and an Erbium laser group (34 cases, 3 cases dropped off) according to the settings. In the electroacupuncture group, electroacupuncture was applied at "four points of sacral region", i.e. points of 0.5 cun beside bilateral sacrococcygeal joints and bilateral Huiyang (BL 35), with continuous wave, 2 Hz in frequency, 60 min each time, once every other day, 3 times a week, 12 times as one course of treatment. In the Erbium laser group, transurethral Erbium laser technology was given, once every 4 weeks as one course of treatment. Both groups were treated for 5 courses. The scores of the International Consultation on Incontinence questionnaire-short form (ICI-Q-SF) and the incontinence quality of life questionnaire (I-QOL) were observed before treatment, after each course of treatment and in follow-up after 1 and 2 months of treatment completion, respectively, and the clinical efficacy was evaluated after treatment in the two groups. RESULTS: Compared with those before treatment, the ICI-Q-SF scores were decreased while the I-QOL scores were increased after 5 courses of treatment and in follow-up after 1, 2 months of treatment completion in the two groups (P<0.01). The ICI-Q-SF score in follow-up after 2 months of treatment completion were higher than that after 5 courses of treatment in the Erbium laser group (P<0.05). After 3, 4, 5 courses of treatment and in follow-up after 1 and 2 months of treatment completion, the ICI-Q-SF scores in the electroacupuncture group were lower than those in the Erbium laser group (P<0.05, P<0.01); after 2, 3, 4, 5 courses of treatment and in follow-up after 1 and 2 months of treatment completion, the I-QOL scores in the electroacupuncture group were higher than those in the Erbium laser group (P<0.01). The change ranges of ICI-Q-SF score and I-QOL score between before treatment and after each course of treatment in the electroacupuncture group were lager than those in the Erbium laser group (P<0.01, P<0.05). The total effective rate was 61.8% (21/34) in the electroacupuncture group, which was superior to 19.4 (6/31) in the Erbium laser group (P<0.01). CONCLUSION Both electroacupuncture at "four points of sacral region" and transurethral Erbium laser can improve the clinical symptoms and the quality of life in patients of moderate to severe stress urinary incontinence after radical prostatectomy. The short-term efficacy and long-term efficacy of electroacupuncture are superior to the Erbium laser technology.
Male ; Humans ; Quality of Life ; Urinary Incontinence, Stress/therapy* ; Sacrococcygeal Region ; Electroacupuncture ; Erbium ; Prostatectomy/adverse effects*

OBJECTIVES: Perfluorooctanoic acid (PFOA) can cause lipid metabolism disorders in animal body and affect the lipolysis and synthesis of fatty acids. Peroxisome proliferators-activated receptor (PPAR) plays an extremely important role in this process. This study aims to explore the effects of PFOA on liver lipid metabolism disorders in Sprague Dewley (SD) rats and the expression of PPAR. METHODS: A total of 40 male SD rats were randomly divided into 4 groups (n=10 in each group): a control group (ddH₂O), a low-dose PFOA group [PFOA 1.25 mg/(kg·d)], a middle-dose PFOA group [PFOA 5.00 mg/(kg·d)], and a high-dose PFOA group [PFOA 20.00 mg/(kg·d)]. The rats were fed with normal diet, and PFOA exposure were performed by oral gavage for 14 days, and the rats were observed, weighted and recorded every day during the exposure. After the exposure, the blood was collected, and the livers were quickly stripped after the rats were killed. Part of the liver tissues were fixed in 4% paraformaldehyde for periodic acid-schiff (PAS) staining; the contents of HDLC, LDLC, TG, TC in serum and liver tissues, as well as the activities of their related enzymes were assayed; The expression levels of cyclic adenosine monophosphate-response element binding protein (Cbp), general control of amino acid synthesis 5-like 2 (Gcn5L2), peroxidation peroxisome proliferation factor activated receptor γ (PPAR), silent information regulator 1 (Sirt1) and human retinoid X receptor alpha 2 (Rxrα2) ) were detected by Western blotting. RESULTS: After 14 days of PFOA exposure, the PAS staining positive particles in the cytoplasm and nucleus of SD rats in the medium and high dose groups were significantly reduced compared with the control group. The serum levels of LDLC and TC in the low-dose and middle-dose groups were significantly reduced compared with the control group (all P<0.05), while the high-dose group showed an increasing tendency, without siginificant difference (P>0.05), there was no significant difference in HDLC and TG (both P>0.05). The activities of alkaline phosphatase (AKP) and alanine aminotransferase (ALT) were increased significantly (both P<0.05) compared with control group; the ratio of ALT/aspartate aminotransferase (AST) in the high-dose group was increased significantly (P<0.05), there was no significant difference in LDH and TG (both P>0.05); the HDLC content in the liver tissues in the high-dose group was significantly reduced, compared with the control group (P<0.05); the TC contents in the liver tissues in the low, medium and high-dose groups were significantly increased (all P<0.05), there was no significant difference in LDLC and TG (both P>0.05); the AKP activity in the livers in the medium and high-dose groups was significantly increased (both P<0.05), there was no siginificant difference in LDH, ALT, and the ratio of ALT/AST (all P>0.05); the protein expression levels of Ppar γ, Cbp and Rxrα2 in the liver in the high dose groups were significantly down-regulated compared with the control group (all P<0.05), while the protein expression levels of Sirt1 were significantly up-regulated (all P<0.05). CONCLUSIONS PFOA exposure can cause lipid metabolism disorder and glycogen reduction in SD rat livers, which may be related to the activation of Sirt1 and inhibition of Ppar γ expression, leading to affecting the normal metabolism of fatty acids and promoting glycolysis.
Animals ; Caprylates ; Fatty Acids/pharmacology* ; Fluorocarbons ; Lipid Metabolism ; Lipid Metabolism Disorders/metabolism* ; Liver/metabolism* ; Male ; PPAR gamma ; Rats ; Rats, Sprague-Dawley ; Sirtuin 1/metabolism*

OBJECTIVETo develop a HPLC method for the simultaneous determination of chlorogenic acid, cryptochlorogenic acid, caffeic acid, naringin, hesperidin and linarin in xiao' erjinning oral liquid.

METHODThe chromatographic separation was achieved on a Lichrospher C18 (4.6 mm x 250 mm, 5 microm) column with a mobile phase which was composed of acetonitrile(A) and 0.4% phosphoric acid(B) for gradient elution (10:90-18:82-27:73). The flow rate was (0.8-1.1-0.8) mL x min(-1), the column temperature was 30 degrees C and the detection wavelength was set at 300 nm.

RESULTThe results showed that 6 effective components were separated well and showed good linearity. The average recoveries were between 95%-105%.

CONCLUSIONThe method is proved to be rapid, accurate, credible and repeatable. It can be used for the quality control of Xiao'erjinning oral liquid.

Caffeic Acids ; analysis ; Chlorogenic Acid ; analysis ; Chromatography, High Pressure Liquid ; methods ; Dosage Forms ; Drugs, Chinese Herbal ; analysis ; Flavanones ; analysis ; Glycosides ; analysis ; Hesperidin ; analysis

In this research, we use mouse embryonic fibroblasts as feeder layers. To eliminate the influence of serum and mouse embryonic stem cells (ESCs) conditioned medium (ESCCM) on self-renewal of sheep embryonic stem-like cells, knockout serum replacement (KSR) was used to replace serum, then supplanted with ESCCM for the isolation and cloning of sheep embryonic stem-like cells. We found when inner cell masses (ICMs) cultured in the control group with medium supplanted with fetal bovine serum (FBS), sheep ES-like cells could not survive for more than 3 passages. However, sheep embryonic stem-like cells could remain undifferentiated for 5 passages when cultured in the medium that FBS was substituted by KSR. The result indicates that KSR culture system was more suitable for the isolation and cloning of sheep embryonic stem-like cells compared to FBS culture system. Finally we applied medium with 15% KSR as basic medium supplanted with 40% ESCCM as a new culture system to isolate sheep embryonic stem-like cells, we found one embryonic stem-like cell line still maintained undifferentiating for 8 passages, which characterized with a normal and stable karyotype and high expression of alkaline phosphatase. These results suggest that it is suitable to culture sheep ICM in the new culture system with 15% KSR as basic medium and supplanted with 40% ESCCM, which indicated that mouse ES cells might secrete factors playing important roles in promoting sheep ES-like cells' self-renewal.
Animals ; Cell Culture Techniques ; methods ; Cell Proliferation ; Cell Separation ; methods ; Clone Cells ; Culture Media ; Embryonic Stem Cells ; cytology ; Mice ; Sheep