Nonalcoholic fatty liver disease （NAFLD） is one of the most prevalent chronic liver diseases in the world， affecting about one quarter of the global population， and it is estimated that NAFLD will become the main indication for liver transplantation by 2030. NAFLD can lead to significant abnormalities in the levels of a variety of amino acids including branched-chain amino acids， thereby promoting the development and progression of NAFLD. These results suggest that in addition to glucose and lipid metabolism， amino acid metabolism also plays an important role in the progression of NAFLD. In order to systematically understand the role and mechanism of amino acid metabolism in NAFLD， this article reviews the research advances in amino acid metabolism in NAFLD. This article aims to explore the role and mechanism of amino acid metabolism in the progression of NAFLD， so as to provide ideas and a theoretical basis for clinical prevention and treatment.

Tropane alkaloids (TAs) are a class of anticholinergic drugs widely used in clinical practice and mainly extracted from plant, among which Atopa belladonna is the main commercial drug source. It is of great industrial value to obtain TAs in large quantities by plant metabolic engineering. In TAs pathway, cytochrome oxidase CYP82M3 catalyze the synthesis of tropinone and then tropinone reductase I (TRI) compete with TRII for tropinone to form tropine leading to the TAs synthesis (drainage). In this study, based on the "increasing flow and drainage" metabolic engineering strategy, two genes, namely HnCYP82M3 and DsTRI from Hyoscyamus niger and Datura stramonium, respectively, were overexpressed in the hair roots of A. belladonna, with a view to promote the TAs accumulation. The HnCYP82M3 gene was cloned from the root of H. niger, and it encoded amino acid with 91.7% sequence identity with AbCYP82M3 from A. belladonna. Overexpression of HnCYP82M3 alone did not affect the content of TAs in hair roots of A. belladonna, indicating that CYP82M3 was not a key enzyme in TAs biosynthesis. Simultaneous overexpression of HnCYP82M3 and DsTRI greatly promoted the accumulation of the three TAs, and the contents of hyoscyamine, anisodamine and scopolamine were 4.97 times, 2.83 times and 2.19 times that of the control, respectively, and the increase amplitude was greater than that of single overexpression of DsTRI. This study showed that the "increasing flow and drainage" strategy of enzyme genes co-expression at branch points was a promising metabolic engineering method to effectively improve the biosynthesis of TAs in A. belladonna, and laid a theoretical and technical foundation for the large-scale industrial acquisition of TAs.

As a serious cardiovascular disease, atherosclerosis (AS) causes chronic inflammation and oxidative stress in the body and poses a threat to human health. Lipoprotein-associated phospholipase A2 (Lp-PLA2) is a member of the phospholipase A2 (PLA2) family, and its elevated levels have been shown to contribute to AS. Lp-PLA2 is closely related to a variety of lipoproteins, and its role in promoting inflammatory responses and oxidative stress in AS is mainly achieved by hydrolyzing oxidized phosphatidylcholine (oxPC) to produce lysophosphatidylcholine (lysoPC). Moreover, macrophage apoptosis within plaque is promoted by localized Lp-PLA2 which also promotes plaque instability. This paper reviews those researches of Chinese medicine in treating AS via reducing Lp-PLA2 levels to guide future experimental studies and clinical applications related to AS.
Humans ; 1-Alkyl-2-acetylglycerophosphocholine Esterase ; Medicine, Chinese Traditional ; Atherosclerosis/drug therapy* ; Lipoproteins ; Plaque, Atherosclerotic ; Biomarkers

BACKGROUND: The effects of acupuncture have varied in different randomized controlled trials (RCTs), and there are many factors that influence treatment effect of acupuncture in different outcomes, with conflicting results. OBJECTIVE: To identify factors and their impact on the treatment effect of acupuncture in different outcomes. METHODS: Acupuncture RCTs were searched from 7 databases including Medline (PubMed), Embase, Cochrane Central Register of Controlled Trials, China National Knowledge Infrastructure, Wanfang Database, VIP Database, and China Biology Medicine disc between January 1st, 2015 and December 31st, 2019. Eligible studies must compare acupuncture to no acupuncture, sham acupuncture, or waiting lists, and report at least 1 patient-important outcome. A multi-level meta-regression was conducted using a 3-level robust mixed model and univariate analyses were performed for all independent variables, even those excluded from the multivariable model due to collinearities. We used thresholds of 0.2 and 0.4 for the difference of standardized mean differences (SMDs), categorising them as small (<0.2), moderate (0.2-0.4), or large (>0.4) effects. RESULTS: The pain construct analysis involved 211 effect estimates from 153 studies and 14 independent variables. High-frequency acupuncture treatment sessions produced larger effects compared to low-frequency sessions [large magnitude, the difference of adjusted SMDs 0.46, 95% confidence interval (CI) 0.07 to 0.84; P=0.02]. The non-pain symptoms construct analysis comprised 323 effect estimates from 231 studies and 15 independent variables. Penetrating acupuncture showed moderately larger effects when compared to non-penetrating acupuncture (0.30, 95% CI 0.06 to 0.53; P=0.01). The function construct analysis included 495 effect estimates from 274 studies and 14 independent variables. Penetrating acupuncture and the flexible acupuncture regimen showed moderately larger effects, compared to non-penetrating acupuncture and fixed regimen, respectively (0.40, 95% CI 0 to 0.80; P=0.05; 0.29, 95% CI 0.06 to 0.53; P=0.01). CONCLUSIONS High-frequency acupuncture sessions appear to be a more effective approach to managing painful symptoms. Penetrating acupuncture demonstrated greater effect in relieving non-painful symptoms. Both penetrating acupuncture type and flexible acupuncture regimen were linked to significant treatment effects in function outcomes. Future studies should consider the factors that are significantly associated with the effects of acupuncture in patient-important outcomes.
Humans ; Randomized Controlled Trials as Topic ; Acupuncture Therapy/methods* ; Pain ; Pain Management ; China

Objective To evaluate the pharmacokinetics and bioequivalence of two entecavir tablets under fasting condition in Chinese healthy volunteers.Methods A single oral dose of 0.50 mg entecavir test and reference formulations was given to 30 healthy volunteers in the fasting state according to two randomized open-label crossover clinical studies.Plasma concentrations were determined by HPLC-MS/MS after deproteinized with acetonitrile.The pharmacokinetic parameters were calculated by WinNonlin 6.4.The bioequivalence of entecavir tablets test and its reference in the fasting state were evaluated respectively.Results In the fasting state,the main pharmacokinetic parameters of entecavir were as follows:Cmax were(4 390.00±1 257.56)and(4 421.00±1 239.55)pg·mL-1;tmax were 0.67(0.33,2.00)and 0.75(0.50,2.00)h;t1/2were(29.71±14.50)and(35.43±16.58)h,AUC0_72h were(12 252.43±2 671.28)and(12 063.43±2 151.50)pg·mL-1·h.The 90％confidence intervals of the mean ratio of Cmax,AUC0-72h and AUC0-∞ of the text and reference formulations were all within the equivalent interval of 80.00％-125.00％.Conclusion The entecavir tablets test and its reference were bioequivalent in the fasting state.

Objective To study the role of sphingosine-1-phosphate(S1P)signal on the proliferation of breast cancer BT549 cells.Methods Cells were divided into control group and experimental group,experimental group were treated with 0.1,1.0,10.0 μmol·L-1 S1P receptor agonist SEW2871 for 72 h.Control group was cultured with 0.1％fetal bovine serum.Cell proliferation was detected by methyl thiazolyl tetrazolium(MTT)assay.Cell models of overexpressing S1P receptors in BT549 were divided into three groups:blank plasmid group(LUC),wild type S1P receptor overexpression group(WT),S1P receptor phosphorylation site mutation overexpression group(MUT);the proliferation ratio was detected by MTT,the number of cell clones was counted by colony formation experiment.S1P antagonist W146(10 μmol·L-1)and protein kinase(AKT)signaling inhibitor MK2206(90 nmol·L-1)were used to detect the role of S1P signaling in the proliferation of breast cancer cells.The expression of phosphorylate signal transducer and activator of transcription 3(p-STAT3),c-Myc proteins were detected by Western blot.Results The growth ratio of BT549 cells in control group and 0.1,1.0,10.0 μmol·L-1experimental groups were 1.00±0.03,1.13±0.06,1.06±0.10 and 1.07±0.03,0.1 μmol·L-1 SEW2871 promot the cell proliferation(P＜0.05).Compared between WT group,MUT group and LUC group,the growth rate and the number of clonal colonies were increased after overexpression of S1P receptor(all P＜0.05).The growth ratio of BT549 cells after treatment with W146 and MK2206 in the LUC group,WT group and MUT group were 1.25±0.12,1.31±0.03,1.43±0.14 and 0.87±0.15,0.77±0.03,0.88±0.02.Compared between MUT group,WT group and corresponding DMSO group,the differences were statistically significant(all P＜0.01).The number of cell clony formation number after treatment with W146 were 65.65±5.12,141.48±5.63 and 93.64±5.14;compared between MUT,WT group and corresponding DMSO group,the differences were statistically significant(all P＜0.05).The relative protein expression levels of p-STAT3 in LUC group,WT group and MUT group were 0.67±0.04,0.69±0.08 and 0.81±0.06,the relative protein expression levels of proto-oncogene c-Myc were 1.69±0.03,0.70±0.10 and 0.67±0.07,compared between WT group,MUT group and corresponding DMSO group,the difference was statistically significant(P＜0.05).Conclusion S1P signaling can promote proliferation in breast cancer BT549 cells,and the mechanism could be related to AKT and STAT3 signaling pathway.

Objective To explore the effects of glycyrrhizic acid(GA)on neurons injury in traumatic brain injury(TBI)rats and the possible mechanism.Methods The rats were randomly divided into sham-operation group,model group,control group and experimental-L,-M,-H groups,with 20 rats each group.The sham-operation group was only treated with craniotomy;the other 5 groups were used to establish TBI models by extracorporeal shock method.At 0,24 and 48 h after modeling,the experimental-L,-M,-H groups were intraperitoneally injected with 10,50 and 100 mg·kg-1 GA solution,respectively;control group was intraperitoneally injected with 2 mg·kg-1 nimodipine;sham-operation and model groups were intraperitoneally injected with the equal volume of phosphate buffered solution.The degree of neurological dysfunction was evaluated by cerebral edema and modified neurological severity score(mNSS).The apoptosis rates of neurons in rat brain tissue was evaluated by apoptosis staining.Western blot was used to analyze the expression levels of apoptosis-related proteins and aquaporin 4(AQP4)protein.Results The mNSS scores of experimental-M,-H groups,control group,model group,sham-operation group were(6.98±0.82),(5.28±0.37),(5.91±0.52),(13.28±1.59)and(0.36±0.01)points;the degrees of brain edema were(63.27±10.33)％,(60.09±9.38)％,(66.86±9.91)％,(85.92±11.93)％and(52.17±8.53)％;the apoptosis rates of neurons were(6.81±0.73)％,(5.39±0.25)％,(5.87±0.62)％,(15.13±3.29)％and(2.56±0.03)％;the relative expression levels of B cell lymphoma 2(Bel-2)protein were 0.49±0.06,0.68±0.15,0.62±0.03,0.13±0.03 and 0.95±0.13;the relative expression levels of Bel-2 associated X protein were 0.61±0.08,0.55±0.17,0.39±0.09,0.92±0.19 and 0.16±0.02;the relative expression levels of AQP4 protein were 0.69±0.15,0.38±0.03,0.47±0.09,0.86±0.13 and 0.13±0.09,respectively.There were statistically significant differences in the above indexes between the model group and the experimental-M,-H groups and control group(all P＜0.05).Conclusion GA is able to reduce the brain edema degree and neurological dysfunction in TBI rats,and inhibit neuronal damage and apoptosis,and the mechanism of action may be associated with the inhibition of AQP 4 expression.

Objective:To summarize the clinical and genetic characteristics of children with β-ketothiolase deficiency (BKTD).Methods:The clinical characteristics, biochemical, markers detected by tandem mass spectrometry (MS/MS) and gas chromatography-mass spectrometry (GC/MS), as well as the variants in ACAT1 gene among 5 children with BKTD in Children′s Hospital of Chongqing Medical University between October 2018 and December 2022 were retrospectively analyzed.Results:The onset age of the disease in 5 patients (4 males and 1 female) ranged from 9.7 to 28.0 months. During the acute phase, severe metabolic acidosis was observed with a pH of 6.9-7.1, as well as hypoglycaemia (2.3-3.4 mmol/L) and positive urinary ketone bodies (+-++++). Blood levels of methylcrotonyl carnitine, methylmalonyl carnitine and malonyl carnitine were 0.03-0.42, 0.34-1.43 and 0.83-3.53 μmol/L respectively and were significantly elevated. Urinary 2-methyl-3-hydroxybutyric acid was 22-202 and 3-hydroxybutyric acid was 4-6 066, both were higher than the normal levels. Methylcrotonylglycine was mild elevated (0-29). The metabolites detected by MS/MS and GC/MS were significantly reduced after treatment. Analysis of ACAT1 gene mutation was performed in 5 children. Most variants were missense (8/9). Four previously unreported variants were identified: c.678G>T (p.Trp226Cys), c.302A>G (p.Gln101Arg), c.627_629dupTGA (p.Asn209_Glu210insAsp) and c.316C>T (p.Gln106Ter), the first 2 variants were predicted to be damaging by SIFT, PolyPhen-2 and Mutation Taster software. c.316C>T (p.Gln106Ter) is a nonsense variant.Conclusions:β-ketothiolase deficiency is relatively rare, lacks specific clinical manifestations, however severe metabolic acidosis, hypoglycemia, and ketosis during the acute onset were consistent findings. Missense mutations in the ACAT1 gene are common genetic causes of β-ketothiolase deficiency.

Carboxylated pillar[5]arene functionalized silver nanoparticles(CP5A-AgNPs)were successfully prepared by Creighton method.The prepared CP5A-AgNPs composites were characterized by ultraviolet-visible absorption spectroscopy(UV-Vis),infrared spectroscopy(FT-IR)and transmission electron microscopy(TEM),etc.TEM results showed that when the molar ratio of CP5A to AgNO3 was 1:10,the prepared CP5A-AgNPs had good dispersion and uniform particle size,with an average particle size of 4.05 nm.The catalytic degradation ability of CP5A-AgNPs toward two kinds of organic dyes,Rhodamine B(RhB)and methyl orange(MO)was further investigated.The results showed that the degradation rates of these two dyes by CP5A-AgNPs were 99.91%and 98.83%respectively,and CP5A-AgNPs exhibited good cyclic catalytic ability.The catalytic efficiencies in the fifth cycle were 91.06%and 98.45%,respectively.In addition,the performance of functionalized silver nanoparticles using monomer compound of CP5(CMA)as stabilizer(CMA-AgNPs)was compared.The results showed that CP5A-AgNPs had strong catalytic degradation activity to RhB and MO,and had good recycling catalytic ability.

Objective To observe the clinical effect of PENG's tendon-separating tuina therapy combined with lateral needling technique for meridian sinew in the treatment of acute scapulohumeral periarthritis(referred to as frozen shoulder),and to provide therapeutic ideas and evidence for the treatment of acute scapulohumeral periarthritis.Methods Sixty patients with acute scapulohumeral periarthritis were randomly divided into treatment group and control group,with 30 cases in each group.The control group was treated with lateral needling technique for meridian sinew,and the treatment group was treated with PENG's tendon-seperating tuina therapy combined with lateral needling technique for meridian sinew.Both groups were treated for once a day,10 times as a course of treatment,and for a total of 2 courses of treatment.The changes of pain Visual Analogue Scale(VAS)score and Constant-Murley Scale(CMS)score of shoulder joint function in the two groups were observed before treatment and after one and two course(s)of treatment.Moreover,the clinical efficacy of the two groups was evaluated.Results(1)After two courses of treatment,the total effective rate of the treatment group was 90.00%(27/30),and that of the control group was 73.30%(22/30).The overall efficacy(tested by rank sum test)and total effective rate(tested by chi-square test)of the treatment group were significantly superior to those of the control group,and the differences were statistically significant(P＜0.05).(2)After one and two course(s)of treatment,the VAS scores of pain in the two groups were significantly decreased than those before treatment(P＜0.05 or P＜0.01);after two courses of treatment,VAS scores in the two groups were significantly lower than those after one course of treatment(P＜0.05).The decrease of VAS scores in the treatment group after one and two course(s)of treatment was significantly superior to that in the control group,and the difference were statistically significant(P＜0.05 or P＜0.01).(3)After one and two course(s)of treatment,the CMS scores of shoulder joint function in the two groups were significantly higher than those before treatment(P＜0.01);after two courses of treatment,the CMS scores in the two groups were higher than those after one course of treatment(P＜0.05).The increase of CMS scores in the treatment group after one and two course(s)of treatment was significantly superior to that in the control group,and the difference was statistically significant(P＜0.01).Conclusion PENG's tendon-separating tuina therapy combined with lateral needling technique for meridian sinew has a definite clinical effect in the treatment of patients with acute scapulohumeral periarthritis,which can effectively relieve the pain and discomfort of shoulder joint and improve the movement function of shoulder joint.