Objective:To investigate the effects and the possible mechanisms of Neiguan(PC6)and Gongsun(SP4)on the hypothalamic-pituitary-adrenal(HPA)axis in rats with functional dyspepsia(FD),thus to provide a theoretical basis for the clinical application of the Eight Confluent Points.Methods:Forty specific-pathogen-free Sprague-Dawley rats were divided into a blank group,a model group,an electroacupuncture(EA)group,and a Western medicine group by the random number table method,with 10 rats in each group.Rats in the blank group did not receive modeling or intervention.Rats in the other three groups were subjected to the FD with mood disorder model using the compound etiology modeling method.After the successful modeling,rats in the model group did not receive any interventions,rats in the Western medicine group received deanxit and mosaprid intervention,and those in the EA group received EA intervention on the ipsilateral Neiguan(PC6)and Gongsun(SP4)for 21 d.The sugar-water consumption rate was measured before the experiment and before and after interventions to assess the emotional status.The gastric emptying rate was measured after interventions to assess the gastrointestinal dynamics.The expression levels of hypothalamic corticotropin-releasing hormone(CRH),pituitary adrenocorticotropic hormone(ACTH),and adrenal corticosterone(CORT)were measured by enzyme-linked immunosorbent assay.Results:Compared with the blank group,the sugar-water consumption rate and the gastric emptying rate were decreased(P<0.01),and the hypothalamic CRH,pituitary ACTH,and adrenal CORT expression levels were increased(P<0.01)in the model group.Compared with the model group,the sugar-water consumption rate and the gastric emptying rate were significantly increased(P<0.01),while the expression levels of hypothalamic CRH,pituitary ACTH,and adrenal CORT were significantly decreased(P<0.01)in the EA group and the Western medicine group.The differences between the EA group and the Western medicine group were not statistically significant(P>0.05).Conclusion:The Eight Confluent Points Neiguan(PC6)and Gongsun(SP4)can improve the mood and gastrointestinal dynamics in FD rats,which may be achieved by down-regulating the hypothalamic CRH,pituitary ACTH,and adrenal CORT,as well as by correcting the HPA axis hyperfunction.

OBJECTIVE To investigate the effects of astaxanthin on oxidative stress and inflammatory reaction in rats with traumatic brain injury （TBI）. METHODS Male SD rats were randomly divided into sham operation group， model group， astaxanthin low-dose group （20 mg/kg）， astaxanthin high-dose group （40 mg/kg）， astaxanthin+ML385 group [astaxanthin 40 mg/kg+ nuclear factor-erythroid 2-related factor 2 （Nrf2） inhibitor ML385 30 mg/kg]， with 14 rats in each group. Except for the sham operation group， TBI model was induced by the modified Feeney free-fall impact method in other groups. The rats in each drug group were given the corresponding drug intragastrically or intraperitoneally， and the rats in the sham operation group and model group were intragastrically given a constant volume of normal saline. The neurological function of rats in each group was scored on the 1st， 3rd and 7th day after drug intervention； on the 7th day of drug intervention， the changes of cerebral histomorphology and inflammatory infiltration score were observed in each group， and the ultrastructure of nerve cells in brain tissue was also observed. The contents of oxidative stress indexes [superoxide dismutase （SOD）， catalase （CAT）， glutathione peroxidase （GSH-Px）， malondialdehyde （MDA）， nitric oxide （NO）] and inflammatory reaction indexes [tumor necrosis factor-α， interleukin-1β （IL-1β）， IL-6， inducible nitric oxide synthase] as well as protein and mRNA expressions of Nrf2， heme oxygenase-1 （HO-1） and NAD（P）H： quinone oxidoreductase 1 （NQO1） were detected in cerebral tissue. RESULTS Compared with the sham operation group， the brain edema of rats in the model group was obvious， accompanied by a large number of inflammatory cells infiltrated， the shape of organelles was damaged and their number was reduced， and the ultrastructure of nerve cells was seriously damaged. The neurological function score， the contents of SOD， CAT， GSH-Px and NO and the relative expression levels of Nrf2， HO-1 and NQO1 protein and mRNA in brain tissue were significantly decreased， while the inflammatory infiltration scores， the contents of MDA and inflammatory reaction indexes were significantly increased （P＜0.05）. Compared with the model group， low-dose and high-dose astaxanthin could significantly improve the pathological status of brain tissue and nerve cells and neurological function scores （except for the first day of drug intervention in the astaxanthin low-dose group）， increase the contents of SOD， CAT， GSH-Px and NO and the relative expression levels of Nrf2， HO-1， NQO1 protein and mRNA in brain tissue in a dose-dependent manner， and reduce inflammatory infiltration scores， the contents of MDA and inflammatory reaction indexes （P＜0.05）. ML385 could significantly inhibit the above effects of astaxanthin （P＜0.05）. CONCLUSIONS Astaxanthin may reduce the oxidative stress of TBI model rats， alleviate the neurological damage and reduce the level of inflammation reaction by activating the Nrf2/HO-1 signaling pathway.

Background and Objectives: This study was to investigate the role of microRNA-29a-3p (miR-29a-3p) in human bone marrow mesenchymal stem cells (hBMSCs), and its relationship with steroid-associated osteonecrosis. Methods: and Results: The online tool GEO2R was used to screen out the differentially expressed genes (DEGs) in GSE123568 dataset. Quantitative real time-polymerase chain reaction (qRT-PCR) was performed to detect the expression of miR-29a-3p, forkhead box O3 (FOXO3), alkaline phosphatase (ALP), bone gamma-carboxyglutamate protein (OCN) and RUNX family transcription factor 2 (Runx2) in the hBMSCs isolated from the patients with steroid-associated osteonecrosis. CCK-8 assay was executed to measure cell viability; western blot assay was utilized to detect FOXO3, ALP, Runx2, OCN and β-catenin expression. Cell apoptosis and cell cycle were detected by flow cytometry. Immunofluorescence assay was used to detect the sub-cellular localization of β-catenin. Bioinformatics analysis and luciferase reporter gene assay were performed to confirm whether miR-29a-3p can combine with FOXO3 3’UTR. MiR-29a-3p was markedly up-regulated in the hBMSCs of patients with steroid-associated osteonecrosis, while FOXO3 mRNA was significantly down-regulated. Transfection of miR-29a-3p mimics significantly inhibited the hBMSCs’ proliferation, osteogenic differentiation markers’ expressions, including ALP, Runx2, OCN, and repressed the ALP activity, as well as promoted cell apoptosis and cell-cycle arrest. FOXO3 was identified as a target gene of miR-29a-3p, and miR-29a-3p can inhibit the expression of FOXO3 and β-catenin, and inhibition of miR-29a-3p promoted translocation of β-catenin to the nucleus. Conclusions MiR-29a-3p can modulate FOXO3 expression and Wnt/β-catenin signaling to inhibit viability and osteogenic differentiation of hBMSCs, thereby promoting the development of steroid-associated osteonecrosis.

Objective:To explore the clinical application value of sound touch elastography (STE) in the evaluation of the curative effect of acupuncture and moxibustion in the treatment of stroke patients with upper limb spasm.Methods:Fifty-five patients with upper limb spasm after stroke were selected as stroke group, and thirty healthy volunteers matched with age, gender and body mass index (BMI) were selected as control group between September 2020 and March 2021. The shear wave velocity (SWV) of biceps brachii and triceps brachii were measured in both groups. In stroke group, the SWVs of biceps brachii and triceps brachii on spastic side and non-spastic side were measured before and after acupuncture treatment, and the correlations between SWV and spastic grade (X), muscle response angle (R1), passive range of motion (R2), spastic angle (R2-R1) of modified Tardieu scale (MTS) were analyzed.Results:At 0 degree, the SWV of spastic biceps brachii (3.85±0.62)m/s was significantly higher than those of the non-spastic side [(3.34±0.35)m/s] and control group [(3.15±2.66)m/s] (all P<0.001), and decreased [ (3.45±0.47)m/s] after acupuncture treatment ( P<0.001). At 90 degree, the SWV of non-spastic biceps brachii was larger than those of spastic side and control group [1.98(1.92, 2.14)m/s vs 1.98(1.92, 2.11) m/s, 1.95(1.86, 2.05)m/s, all P<0.05], and there was no significant difference between the SWV of spastic side and control group ( P>0.05). There were no significant differences in SWV of triceps brachii between the spastic side and control group at both 0 degree or 90 degree ( P>0.05). At 0 degree, SWV positively correlated with MTS (x) ( rs=0.639, P<0.001) and negatively with R1 ( rs=-0.527, P<0.001) and R2 ( rs=-0.825, P<0.001) on the spastic biceps brachii. Conclusions:STE can provide quantitative index for the objective evaluation of the curative effect of acupuncture in upper limb spasm after stroke, and it is proposed to be an effective means of clinical diagnosis and efficacy evaluation in patients with upper limb spasm after stroke.

Objective To investigate the influence of nonalcoholic fatty liver disease (NAFLD) on the antiviral response of patients with chronic hepatitis B (CHB), and to provide a reference for clinical treatment of such patients. Methods A total of 187 patients who attended Shenzhen Third People's Hospital from January 2011 to December 2017 were enrolled and divided into CHB group with 43 patients, NAFLD group with 41 patients, and CHB+NAFLD group with 103 patients. Related indices were measured at enrollment different time points of follow-up, including body height, body weight, alanine aminotransferase (ALT), aspartate aminotransferase, four blood lipid parameters, four indicators of liver fibrosis, aspartate aminotransferase-to-platelet ratio index, HBsAg, HBeAg, anti-HBe, and HBV DNA quantification, and the CHB patients and the CHB+NAFLD patients receiving antiviral therapy were compared in terms of treatment outcome at weeks 12, 24, 48, 72, and 96 of antiviral therapy. The Kruskal-Wallis H test was used for comparison of non-normally distributed continuous data between multiple groups, and the Wilcoxon rank-sum test was used for comparison between two groups; the chi-square test was used for comparison of categorical data between groups. Results Compared with the NAFLD group at baseline, the CHB group and the CHB+NAFLD group had significantly lower platelet count, ALT, gamma-glutamyl transpeptidase (GGT), alkaline phosphatase, and right lobe of liver oblique diameter (all P < 0.05), and compared with the CHB group, the CHB+NAFLD group had significantly higher body mass index, total cholesterol, and triglyceride and a significantly lower spleen thickness (all P < 0.05), while there were no significant differences in the other indicators between the two groups at baseline (all P > 0.05). At week 12 of antiviral therapy, there were no significant differences in liver fibrosis markers and inflammatory indices between the CHB group and the CHB+NAFLD group (all P > 0.05); compared with the CHB+NAFLD group at weeks 24 and 48, the CHB group had significantly greater reductions in ALT ( Z =-2.128 and -3.055, both P < 0.05) and GGT ( Z =-2.025 and -1.631, both P < 0.05); at week 48, the CHB group and the CHB+NAFLD group had a significant reduction in HBV DNA ( Z =-6.445 and -4.415, both P < 0.001), and the CHB group had a significantly greater reduction. The CHB+NAFLD group had a significantly lower HBV DNA clearance rate than the CHB group at different time points of antiviral therapy ( χ 2 =14.237, 13.961, 15.226, 10.462, and 13.030, all P < 0.05). At week 48 of antiviral therapy, the CHB+NAFLD group had a significantly lower HBeAg clearance rate than the CHB group ( χ 2 =5.309, P =0.021), while there was no significant difference between the two groups at week 96 ( χ 2 =0.117, P =0.732). At weeks 24, 48, 72, and 96 of antiviral therapy, the CHB+NAFLD group had a significantly lower ALT normalization rate than the CHB group ( χ 2 =12.049, 5.287, 11.407, and 11.375, all P < 0.05). Conclusion NAFLD reduces the antiviral response of CHB patients and prolongs the duration of antiviral therapy.

Objective:To study the effect of combined decoction and single decoction of Houpu Wenzhongtang on rats with deficiency-cold of spleen and stomach from the perspective of metabonomics, to find out the relevant potential biomarkers and metabolic pathways, and to explore the similarities and differences between the combined decoction and single decoction, so as to provide reference for the feasibility analysis of replacing traditional decoction with single dispensing granule of this formula. Method:SD rats were randomly divided into normal group, model group, Houpu Wenzhongtang combined decoction group and simgle decoction group. Rats in the normal group were given distilled water by intragastric administration, rats in the other three groups were given cold vinegar at 4 ℃ in the morning and refined lard in the afternoon for 10 days (the dosage of 10 mL·kg^-1). After the model was successfully established, rats in the combined decoction group and the single decoction group were given corresponding decoction with dosage of 1.8 g·kg^-1 (according to the amount of crude drugs), once a day for 7 days. Ultra-high liquid chromatography-quadrupole-time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MS) technique was used to analyze the small molecular endogenous metabolites in urine. Orthogonal partial least squares-discriminant analysis (OPLS-DA) and principal component analysis (PCA) were used to compare the changes of differential metabolites among the normal group, model group, Houpu Wenzhongtang combined decoction group and single decoction group, and the differential metabolites were introduced into Kyoto Encyclopedia of Genes and Genomes (KEGG) for metabolic pathway analysis. Result:Compared with the model group, the Houpu Wenzhongtang combined decoction group and single decoction group jointly regulated 13 potential biomarkers, including phosphatidylcholine(PC), lysophosphatidic acid(LysoPA) and cholic acid, etc. They played a role in treating deficiency-cold of spleen and stomach by influencing metabolic pathways such as glycerophospholipid metabolism, glycerolipid metabolism, phosphatidylinositol signaling system and so on. The combined decoction and single decoction of Houpu Wenzhongtang could obviously restore the body weight, motilin and gastrin contents of rats with deficiency-cold of spleen and stomach to normal levels. Conclusion:According to biochemical indexes, there is no obvious difference between combined decoction and single decoction of Houpu Wenzhongtang, but according to metabonomics, the combined decoction may be slightly better than the single decoction. The research shows that it is feasible to replace traditional decoction with single dispensing granule of Houpu Wenzhongtang in clinical application.