Objectives To investigate whether degradation of anterior pharynx decfective-1(Aph-1) goes through proteasomal pathway or lysosomal pathway.Methods Various methods such as cell culture,Western blotting,pulse-chase metabolic labeling technique,double immunofluoresecnt staining,combined with proteasomal and lysosomal inhibition were used to check Aph-1 expression level in stable Aph-1-transfected or non-transfected neuronal(SH-SY5Y)cell line.Results Using Western blotting,treating the neuronal cells with proteasome specific inhibitors significantly increased the expression of both endogenous and exogenous Aph-1.The effect of the proteasome inhibitors on Aph-1 expression was dose-and time-dependent Lysosomal pathway was not involved in Aph-1 degradation. Pulse-chase metabolic labeling experiment showed that the turnover of newly-synthesized radiolabeled Aph-1 protein was blocked by Lactacystin.Double immunofluorescent staining revealed colocalization of Aph-1 and ubiquitin in the same cells.Conclusion Degradation of Aph-1 protein is mediated by proteasomal pathway in neuronal cells,and is not related to lysosomal pathway.Aph-1 protein is ubiquitinated before degradation.

OBJECTIVEThe purpose of this study is to investigate the synergistic effects of human tumor necrosis factor-alpha (hTNF-alpha) transfection and interferon-gamma (IFN-gamma) on the growth of tongue carcinoma cells.

METHODSThe cultured Tca8113 tongue carcinoma cells was divided into 2 groups, one group was transferred with hTNF-alpha gene. Each of the 2 groups was then divided into 5 subgroups, and the subgroups were added IFN-gamma until the final IFN-gamma concentrations respectively were 0, 10, 100, and 1000 U/ml. After culturing for 48 hours, the survival rates of the all groups of cells were assayed by MTT enzymatic labeling technique, and the expression of hTNF-alpha in Tca8113 cells was observed with immunocytochemistry.

RESULTSIFN-gamma did not affect the growth of Tca8113 cells without hTNF-alpha, however, the transfection of hTNF-alpha with the above different concentrations of IFN-gamma synergistically inhibited the growth of Tca8113 cells, the concentrations of IFN-gamma were positively correlated with the inhibition effects (r = 0.733, P < 0.01), the transferred Tca8113 cells displayed remarkable overexpression of hTNF-alpha, compared with the non-transferred.

CONCLUSIONIFN-gamma can enhance the inhibition of hTNF-alpha transfection on the tongue carcinoma cells.

Carcinoma, Squamous Cell ; genetics ; pathology ; Cell Division ; drug effects ; Cell Survival ; drug effects ; Dose-Response Relationship, Drug ; Drug Synergism ; Genetic Vectors ; Humans ; Interferon-gamma ; pharmacology ; Plasmids ; genetics ; Tongue Neoplasms ; genetics ; pathology ; Transfection ; Tumor Cells, Cultured ; Tumor Necrosis Factor-alpha ; genetics

OBJECTIVEThe aim of this study was to investigate suppression effects of the transfection of human tumor necrosis factor-alpha (hTNF-alpha) gene on tongue carcinoma cells.

METHODSThe shuttle plasmid containing hTNF-alpha gene was extracted and purified, then it was transferred into Tca8113 tongue carcinoma cells with cationic liposome DOSPER. The control group was only given equivalent liposomes, except the plasmid. After culturing for 24, 48, 72 and 96 hours, the expression of hTNF-alpha gene in Tca8113 cells was analysed by ELISA and, the survival rate of transferred cells was assayed by MTT enzymatic labeling technique.

RESULTSThe transferred Tca8113 cells displayed significantly overexpression of hTNF-alpha (P < 0.05). The survival rate of the transferred Tca8113 cells was decreased significantly (P < 0.05).

CONCLUSIONTransfection of hTNF-alpha gene in vitro mediated by cationic liposomes can induce the overexpression of hTNF-alpha and inhibit the growth of tongue carcinoma cells.

Carcinoma, Squamous Cell ; genetics ; pathology ; Cell Division ; Gene Transfer Techniques ; Genetic Vectors ; Humans ; Plasmids ; genetics ; Tongue Neoplasms ; genetics ; pathology ; Transfection ; Tumor Cells, Cultured ; Tumor Necrosis Factor-alpha ; genetics

Subthalamic nucleus (STN) deep brain stimulation (DBS) has become an important surgical treatment of Parkinson disease, but its exact mechanism is still unclear.In this study, a 16-channel implantable microelectrode array (MEA) was prepared by micro-electromechanical system (MEMS) technique and later modified with platinum black/reduced Graphene Oxide/Nafion (Pt/RGO/Nafion) nanocomposites.Extracellular dopamine (DA) content and spike of dorsal striatum neurons were synchronously recorded before and after STN stimulation.The results showed that the dopamine content began to increase within 20 s after electrical stimulation and dropped to normal level after about 50 s, with the highest rising concentration of 1.72 μmol/L.At the same time, there was an increased spike activity of interneurons in the dopamine ascending phase, and the spike firing rate of medium spiny projection neurons (MSNs) was high when the concentration of DA was higher than the normal level.The MEA sensor can simultaneously record dopamine flux and physiological signals in situ, thus providing an ideal tool for neural information detection.

ObjectiveTo investigate the protective effect of fecal microbiota transplantation (FMT) on mice with acute-on-chronic liver failure (ACLF) and its effect on intestinal flora. MethodsA total of 40 mice were randomly divided into control group (CON group), model group (MOD group), FMT group (feces of the mice in the CON group were used as fecal microbiota donor), and FMT model group (ANFMT group, with feces of the mice in the MOD group as fecal microbiota donor), with 10 mice in each group. All mice were observed in terms of body weight, death, liver histopathology, and changes in aspartate aminotransferase (AST), alanine aminotransferase (ALT), and intestinal flora. A one-way analysis of variance was used for comparison of normally distributed continuous data between multiple groups, and the SNK-q test was used for further comparison between two groups. ResultsCompared with the CON group, the MOD group had a significant reduction in body weight and significant increases in AST and ALT (all P＜0.05), as well as large patchy necrosis of hepatocytes, significant increases in Verrucomicrobia, Akkermansia, and Erysipelatoclostridium, and significant reductions in Dubosiella and Duncaniella (all P＜0.05). Compared with the CON group, the ANFMT group had a significant increase in AST (P＜0.05), hepatocyte swelling and mild ballooning degeneration, significant increases in Unclassified and Faecalibaculum, and significant reductions in Patescibacteria, Deferribacteres, Muribaculum, Candidatus_Saccharimonas, Rikenella, Odoribacter, Mucispirillum, and Lachnospiraceae_unclassified (all P＜0.05). Compared with the MOD group, the FMT group had significant reductions in AST and ALT (both P＜0.05), mild hepatocellular necrosis and marked ballooning degeneration, significant increases in Paramuribaculum and Bilophila, and significant reductions in Firmicutes, Rikenella, and Absiella (all P＜0.05). ConclusionIntestinal flora disturbance is observed in ACLF mice, and dysbacteriosis may lead to liver injury. FMT can alleviate liver inflammation in ACLF mice and thus exert a protective effect.

Objective To evaluate the clinical efficacy and hemorheological effects of traditional Chinese medicine injection combined with conventional Western medicine in the treatment of stable angina pectoris in coronarrt heartdisease using a mesh meta-analysis system.Methods Computer searches were conducted on China National Knowledge Infrastructure(CNKI),WanFang Data Knowledge Service Platform(WanFang Data),VIP,SinoMed,PubMed,Cochrane Library,Web of Science,and other clinical randomized controlled trials(RCTs)related to the combination of traditional Chinese medicine injection and conventional Western medicine for the treatment of stable angina pectoris in coronary heart disease.The search was conducted until April 2023.The Cochrane bias risk assessment tool was used to evaluate the quality of the included studies,and Stata13.0 software was used for meta-analysis.Results 57 RCTs were ultimately included,involving 5 types of traditional Chinese medicine injections.The results of the network meta-analysis showed that:① in terms of clinical total effective rate,the efficacy of traditional Chinese medicine injection was ranked in the following order:conventional Western medicine treatment combined with Shenmai injection>Shenxiong glucose injection>Shenfu injection>Ciwujia injection>Huangqi injection>conventional Western medicine treatment;② In terms of the effective rate of angina symptoms,the order of efficacy of traditional Chinese medicine injection is conventional Western medicine treatment combined with Shenxiong glucose injection>Ciwujia injection+conventional therapy>Huangqi injection>Shenfu injection>Shenmai injection>conventional Western medicine treatment;③ In terms of the effective rate of electrocardiogram,the order of efficacy of traditional Chinese medicine injection is conventional Western medicine treatment combined with Shenmai injection>Shenxiong glucose injection>Huangqi injection>Ciwujia injection>Shenfu injection>conventional Western medicine treatment;④ In terms of improving the high shear viscosity of whole blood,the efficacy of traditional Chinese medicine injection is ranked in the following order:conventional Western medicine treatment combined with Shenmai injection>Shenxiong glucose injection>Shenfu injection>Huangqi injection>Ciwujia injection>conventional Western medicine treatment;⑤ In terms of improving whole blood low shear viscosity,the efficacy of traditional Chinese medicine injections is ranked in the following order:conventional Western medicine treatment combined with Shenfu injection>Shenmai injection>Shenxiong glucose injection>Ciwujia injection>Huangqi injection>conventional Western medicine treatment;⑥ In terms of improving plasma viscosity,the order of efficacy of traditional Chinese medicine injection is conventional Western medicine treatment combined with Huangqi injection>Shenfu injection>Shenmai injection>Shenxiong glucose injection>Ciwujia injection>conventional Western medicine treatment;⑦ In terms of improving fibrinogen,the efficacy ranking of traditional Chinese medicine injection is conventional Western medicine treatment combined with Shenfu injection>Shenxiong glucose injection>Shenmai injection>Ciwujia injection>conventional Western medicine treatment.Conclusion The combination of traditional Chinese medicine injection and conventional Western medicine treatment has a significant effect on stable angina pectoris in coronary heart disease.Among them,Shenmai Injection has the best therapeutic effect in terms of clinical total effective rate,electrocardiogram,and improvement of whole blood high shear viscosity;In terms of improving angina symptoms,Shenxiong Glucose Injection has the best effect;In terms of improving whole blood low shear viscosity and fibrinogen,Shenfu Injection has the best effect;In terms of improving plasma viscosity,Shenfu Injection has the best effect,but currently it is limited by the number and quality of studies included,and the above conclusions still need to be verified by more high-quality studies.