AIMTo investigate whether free fatty acid impair insulin signaling proteins to inhibit insulin action in rat islet cells.

METHODSAfter rat islet cells were incubated with palmitate (0.25 mmol/L) or oleate (0.125 mmol/L) for 12 hours, 24 hours and 36 hours, western bolt was used to assess the protein abundance of cPKCalpha, Grb2 and FERK2.

RESULTSThe protein content of cPKCalpha were significantly upregulated and the protein abundance of Grb2 and ERK2 was decreased comparing with control in rat islet cells after incubated with free fatty acids.

CONCLUSIONSFree fatty acids may inhibit insulin activity in rat islet cells through up-regulating the expression of cPKCalpha or down-regulating the expression of Grb2 and ERK2.

Animals ; Cells, Cultured ; Fatty Acids, Nonesterified ; blood ; GRB2 Adaptor Protein ; metabolism ; Insulin ; metabolism ; Islets of Langerhans ; metabolism ; Mitogen-Activated Protein Kinase 1 ; metabolism ; Protein Kinase C-alpha ; metabolism ; Rats ; Rats, Sprague-Dawley ; Signal Transduction

AIM: To investigate the clinical effect for treating bullous keratopathy ( BK ) by anterior corneal stromal acupuncture combined with amniotic membrane transplantation.
METHODS: Totally 35 patients ( 35 eyes ) with bullous keratopathy were treated by corneal stromal acupuncture combined with amniotic membrane transplantation. All patients preoperative and postoperative underwent anterior segment OCT, corneal topography, corneal sensitivity and confocal microscopy. To observe postoperative ocular symptoms in patients with BK, recurrence of bulla, changes in corneal thickness, corneal sensitivity and changes in the organizational structure of the layers of the cornea.
RESULTS: Thirty-five were followed up for 6-18mo. The symptom of pain disappeared in 32 cases ( 91%) in the first day after operation and did not recur during follow-up. The symptom of pain relieved in 3 cases (9%) in the first day after operation and disappeared in 3 days. Corneal epithelium of 9 cases ( 26%) were all healed within 1wk, 21 cases (60%) were all healed within 2wk, and 5 cases (14%) were all healed within 3wk. Following up for 6-18mo, there was no recurrence of symptoms or bulla. A small amount of tiny bubbles in the surrounding area appeared in 2 cases after 3 and 4wk. All patients had no neovascularization, and had smooth corneal surface. The amnions of 30 cases (86%) were thinning after 2mo, partially dissolved and absorped, not seen with the naked eye after 3mo. Thirty-four cases (97%) had no changes in vision, one case (3%) was from the light to front of the manual. After 2mo, corneal sensation decreased in 30 cases ( 86%) , corneal thickness increased from preoperative 788±35μm to 940±43μm. After 12mo, corneal thickness increased to 1060±27μm. Results of confocal microscopy: after 3mo, the number density of the trigeminal nerve fibers under corneal basement membrane reduced, shallow stromal cells became into fibrotic stroma, deep stroma was more loose, and cells swelled significantly. The number of endothelial cells reduced and form swelled more obviously compared with preoperatively.
CONCLUSION: Corneal acupuncture combined with amniotic membrane transplantation can effectively control the symptoms of BK, prevent the recurrence of BK, and especially it is a simple, safe and practical way for patients with poor visual function.

Objective To investigate the levels of trace elements such as fluorine(F), and aluminium (Al)etc. of osteomalacia malformation children and to make etiological diagnosis in reference with clinical manifestations.Methods Urine and occipitalia hairs of 14 diseased children(patient group) from endemic fluorosis area and 13 healthy children(control group) from non-endemic area were included in the study on November, 2008, and contents of 10 elements of fluorine(F), aluminum(Al), chromium(Cr), manganese(Mn), ferrum(Fe), cuprum(Cu), zinc(Zn), arsenic (As), selenium(Se), strontium(Sr), and barium(Ba) were tested. The data were analyzed with medical soft package PEMS 3.1. Results Urinary contents of F, Al, Mn, Cu, Sr, and Se(1.18 mg/L, 112.6 μg/L,6.62,29.86 mg/L, 177.5,4.23 ng/L) in patient group were significantly different from those in control group (0.48,47.1,2.04,16.61 mg/L, 55.17,15.52 ng/L, t = 4.592,2.486,4.850,2.210 2.078,2.912, all P＜ 0.05); Hair contents of Al, Mn, As, Sr, Ba, Fe, and Se in patient group(59.27,5.26,0.96,1.50,1.29,297.13,0.45 mg/kg)were significantly different from those of control group( 18.69,0.72,1.09,0.62,0.68,69.02,1.323 mg/kg, t = 4.583,6.318,3.309,2.704,5.606,6.294, all P ＜ 0.05); in patient group, the correlation coefficients of urinary Fe to Al,Zn, As, and Se were all bigger tan 0.662(all P＜ 0.05), those of urinary Se to Mn, Ba, Cu, Zn, Sr, and As were all bigger than 0.694(all P＜ 0.05), those among urinary Mn, Sr, As, and Ba were bigger than 0.550(all P＜0.05), those of hair Al to Mn, Cr, Fe, and Cu were bigger than 0.732(all P＜ 0.05), those of hair Ba to Mn,Cr, Fe, and Sr, and of hair Mn to Cr and Fe, and those between Cr and As, between Cu and Sr were all bigger than 0.686 (all P ＜ 0.05). In control group, the correlation coefficients of urinary Cu to Zn, Se, and Ba, those of Zn to Se and Ba, and those of Cr to Mn and Ba were all bigger than 0.516(all P ＜ 0.05), those of hair Al to Mn,Fe, Cu, As, and Se, and those of hair Se to Fe, Cu, and As, those of hair Fe to Mn, Cu, and As, those of hair Cu to Zn and As, and that between Zn and As were bigger than 0.739(all P ＜ 0.05). The correlation coefficient of urinary F to Se in patient group(0.762) was significantly different from that in control group( - 0.469, u = 2.079,P ＜ 0.05). Conclusions The burden of F and Al of osteomalacia malformation children in endemic fluorosis area of Shuicheng county is too high. The contents of multi-elements in urine and hairs and their correlation are coincident with high levels of Al and F and they cause network increase of multi-element content changes and their correlation. According to bone X-ray features combining with the living environment, the diagnosis of endemic Al-F fluorosis can be made. The biological significance of reducing urinary and hair Se levels and the correlations of F and Al need to be further studied.

A strain of P. Aeruginosa,which was seperated from clinical environment,shows a special characteristic. It keeps normal short rod shape when cultured at 37℃, however,it forms filament without pyocyanin producing when cultured at 25℃ overnight. The filaments will divide and form short rods, simultaneously, produce pyocyanin when culture time is prolonged to over 72h or culture temperature is raised to 37℃. The preliminary study indicates that this phenomenin has nothing to do with nutritive conditions and could the inbluenced by inoculating density and irradiating with ultraviolet rays The absence of pyocyanin was not the cause of filamentous formation by the test results.

Objective To discuss a optimal culture method of primary hippocampal neurons and a more suitable method of mor-phological observation ,and provide basis to the study of synapse in Alzheimer′s Disease .Methods Postnatal 0 -1 days (P0 -1 ) C57BL/6J mice were decollated and bilateral hippocampus were separated .Low level concentration of trypsin and mechanical disso-ciation were adopted .And culture medium without serum was used to culture neurons .After 17 days culturing ,transfected neurons with Green Fluorescent Protein(GFP) by calcium phosphate precipitation ,and then observed neurons and spines by fluorescence mi-croscope .Results The neurons looked good and healthy by using this method .And the axons ,dendrites and spines which were typ-ical structure of neurons were observed clearly after transfected with GFP .Conclusion The cultured hippocampal neurons look good by this method .And the morphological characteristics of neurons and spines are observed much more clearly after transfected GFP by calcium phosphate precipitation .

This study aims to investigate the serotype distribution of non-polio enterovirus (NPEV) isolated from patients with acute flaccid paralysis (AFP) during 2011-2012 in Hebei Province, China and to analyze the relationship between these viruses and AFP. NPEV strains were isolated from the stool specimens from AFP cases in Hebei using human rhabdomyosarcoma cells (RD) and the mouse cell line expressing the gene for the human cellular receptor for poliovirus (L20B) according to the WHO requirements. The nucleotide sequence of VP1 region was determined, and the serotypes of NPEV were identified by molecular typing. The results showed that among the 82 strains of NPEV isolated from the AFP cases during 2011-2012, 42 isolates (55.3%) were identified as human enterovirus A (HEV-A), which were classified into 4 serotypes, 34 (44.7%) as human enterovirus B (HEV-B), which were classified into 13 serotypes, 2 as adenovirus, and 4 were untyped; human enteroviruses C and D were not found in these cases. Enterovirus A71 (EV-A71) was the main type of HEV-A, accounting for 85.7% of all HEV-A strains. HEV-A, especially EV-A71, was predominant among the NPEV strains isolated from AFP patients during 2011-2012 in Hebei Province.
Acute Disease ; China ; epidemiology ; Enterovirus ; classification ; physiology ; Humans ; Paralysis ; epidemiology ; virology ; Seasons ; Serotyping

OBJECTIVETo study the clinical effects of Yupei Qisun Sequential Method (YQSM, by Shen supplementing and Pi invigorating) of Chinese medicine on correlated indices of repeated implantation failure patients in the fresh cycle.

METHODSSixty patients with more than three failure cycles of in virto fertilization and embryo transfer (IVF-ET) or intracytoplasmic sperm injection (ICSI) were recruited. They were assigned to the treatment group (treated by IVF/ICSI and Chinese medicine) and the control group (treated by IVF/ICSI alone), 30 in each group. The total dose of gonadotropin (Gn), the days of controlled ovary hyperstimulation (COH), the thickness of endometrium on the day of embryo transplantation, the number of retrieved oocytes, the fertilization number, the embryo number, the high quality embryo number, the pregnancy rate, and the implantation rate were compared.

RESULTSIn the treatment group the numbers of embryo and high quality embryo were 7.5 +/- 4.9 and 5.1 +/- 3.2 respectively, which were both higher than those of the control group with significant difference (5.1 +/- 3.2, 3.2 +/- 1.8; P < 0.05). No significant difference existed in aspects of the total numbers of Gn, the days of COH, the thickness of endometrium on the day of embryo transplantation, the numbers of retrieved oocytes, the fertilization number, the pregnancy rate, and the implantation rate between the two groups (P > 0.05).

CONCLUSIONYQSM combined with COH could improve the quantity and the quality of embryos, which was promising to increase the accumulative pregnancy rate of RIF patients.

Adult ; Embryo Transfer ; Endometrium ; physiology ; Female ; Fertilization in Vitro ; Gonadotropins ; administration & dosage ; Humans ; Medicine, Chinese Traditional ; methods ; Oocyte Retrieval ; Ovulation Induction ; Pregnancy ; Pregnancy Rate ; Sperm Injections, Intracytoplasmic ; Treatment Failure

In this study, we explored the mechanism of Huganning tablet (HGNP) in the treatment of nonalcoholic fatty liver disease (NAFLD) based on network pharmacology and computer-aided drug design. Firstly, the potential ingredients and targets of HGNP were identified from TCMSP database, Swiss Target Prediction database, Chinese pharmacopoeia (2015) and literatures, and then the targets of HGNP intersected with NAFLD disease targets that obtained in GeneCards database to acquired potential targets. The bioconductor bioinformatics package of R software was used for gene ontology (GO) enrichment and Kyoto encyclopedia of genes and genomes (KEGG) enrichment analysis. The network of “potential ingredient-key target-pathway” was formed in Cytoscape software to study the interactions between potential ingredients of HGNP, key targets, pathways and NAFLD. Based on the results of network pharmacology, the molecular docking analysis of the key targets and potential active ingredients in HGNP tablets with top degree in the network was conducted using Discovery Studio 2020 software, followed by molecular dynamics simulations, binding free energy calculation, drug-likeness properties analysis and ADMET (absorption, distribution, metabolism, excretion and toxicity) properties prediction. In vitro, HepG2 cells were used to establish steatosis model, and the effects of five key compounds on hepatocyte steatosis were analyzed by oil red O staining and triglyceride (TG) content determination. The results showed that 141 ingredients and 151 potential targets were obtained. A total of 2 526 items and 151 pathways were identified by GO and KEGG enrichment analysis. The molecular docking suggested that five components, isorhamnetin, salvianolic acid B, emodin, resveratrol and rhein, exhibited strong binding ability with key targets [retinoic acid receptor RXR-alpha (RXRA), tumor necrosis factor (TNF), glycogen synthase kinase-3 beta (GSK3B), serine/threonine-protein kinase 1 (AKT1)]. It was further verified that isorhamnetin and salvianolic acid B bind to key targets with good structural stability and binding affinity based on molecular dynamics simulations and binding free energy calculations. The drug-likeness properties, pharmacokinetic properties and toxicity of five key compounds were more comprehensively analyzed through drug-likeness properties analysis and ADMET properties prediction. In vitro, all five compounds, isorhamnetin, salvianolic acid B, emodin, resveratrol, and rhein, improved hepatocyte steatosis of HepG2 cells, confirming the reliability of the present study. In conclusion, based on network pharmacology, computer-aided drug design and in vitro validation, this study investigated the mechanism of HGNP for the treatment of NAFLD at multiple levels and provided a basis for its clinical application.

OBJECTIVESTo investigate the clinical effect of sequential therapeutic intervention Yupei Qisun [compensating for weakness by invigorating Kidney (Shen) and Spleen (Pi) in advance] in Chinese medicine (CM) and hysteroscopic endometrial mechanical stimulation on the treatment of infertile patients with repeated implantation failure (RIF); and to study the differences in patients' endometrial thickness and type on the day of embryo transfer, serum hormone levels on embryo transfer day and clinical pregnancy outcomes.

METHODSIn the clinical study, 168 frozen-thawed embryo transfer (FET) cycles for couples with RIF conforming to the research protocol were randomly divided into three groups: a CM group with 56 cycles (CM combined with FET), a hysteroscopy group with 55 cycles (hysteroscopic endometrial mechanical stimulation), and a control group with 57 cycles (conventional FET). Differences in endometrial thickness on the embryo transfer day, levels of serum estradiol (E2) and progesterone (P) on the embryo transfer day, the E2/P ratio on the embryo transfer day, biochemical and clinical pregnancy rates, implantation rate, abnormal pregnancy rate and other indices were compared among the three groups.

RESULTSEndometrial thickness, E2 and P levels, and the E2/P ratio on embryo transfer day and other factors had no significant differences among groups. The biochemical pregnancy, clinical pregnancy, and implantation rates of the CM and hysteroscopy groups were significantly higher than the control group (P<0.05), and there were no significant differences between these two groups. The abnormal pregnancy rate had no significant difference among the three groups.

CONCLUSIONSSequential therapy of Yupei Qisun could significantly improve the clinical outcomes of rif-fet cycles, being equivalent to hysteroscopic endometrial mechanical stimulation, and provided a reliable method to treat such infertile couples.

Abortion, Habitual ; therapy ; Adult ; Embryo Implantation ; Embryo Loss ; therapy ; Embryo Transfer ; Endometrium ; pathology ; physiopathology ; Female ; Humans ; Hysteroscopy ; Infertility, Female ; pathology ; therapy ; Medicine, Chinese Traditional ; methods ; Physical Stimulation ; methods ; Pregnancy ; Retreatment ; statistics & numerical data

AIMTo prepare fluorescein sodium (FS) cationic liposomes and investigate the influence of cationic lipid (DC-chol) and polyethylene glycol (PEG) with different molecule weight (MW) on cationic liposome incorporation efficiency, cellular delivery and fluidity of liposome membrane.

METHODSUsing FS as a model material for encapsulation, the liposomes were prepared and separated (by sephadex G-50 1 cm x 20 cm column), and the liposome incorporation efficiencies was measured. The interaction between the FS and cationic liposomes was investigated by measuring the change of fluorescent spectrum. The cellular uptake of different liposome forms by choosing HepG2 2.2.15 as an in vitro cell culture assay model, and the influence of PEG on the fluidity of liposome membrane with the technique of fluorescence polarization were investigated.

RESULTSCationic lipid and different PEGs showed great effects on increasing liposome incorporation efficiency (from 0.64% to 86.57%), cellular uptake (from 2.18% to 48.46%) and fluidity of liposome membrane. The effect of PEG was MW dependent, and with the increase of MW, the incorporation efficiency and transfection was improved, and the fluidity of liposome membrane increased.

CONCLUSIONAddition of cationic lipid and high MW PEG into cationic liposomes can enhance the cellular delivery and fluidity of cationic liposomes. Also, they can improve the incorporation efficiency of cationic liposomes.

Cholesterol ; analogs & derivatives ; pharmacology ; Drug Delivery Systems ; Fluorescein ; Hepatoblastoma ; pathology ; Humans ; Liposomes ; Liver Neoplasms ; pathology ; Membrane Fluidity ; drug effects ; Molecular Weight ; Polyethylene Glycols ; chemistry ; pharmacology ; Transfection ; methods ; Tumor Cells, Cultured