ObjectiveTo investigate the interventional effects of Shugan Jianpi Yangxin prescription on the expression of orexin-A （OXA）， orexin-1 receptor （OX1R）， and orexin-2 receptor （OX2R） in the mouse model of insomnia. MethodThe mouse model of insomnia was established by intraperitoneal injection of DL-4-chlorophenylalanine （PCPA）. Fifty BALB/c mice were randomized into a blank group， a model group， an eszopiclone （0.13 mg·kg^-1） group， and low- and high-dose （8.4 and 33.6 g·kg^-1， respectively） Shugan Jianpi Yangxin prescription groups and treated with the corresponding drugs for 14 consecutive days. The weight changes of mice were monitored， and Morris water maze and pentobarbital-induced sleep tests were conducted. Immunohistochemistry （IHC） was employed to examine the expression of OXA in the hypothalamus. Enzyme-linked immunosorbent assay was used to measure the levels of OXA and 5-hydroxytryptamine （5-HT） in the hypothalamus， serum， and spleen. Real-time fluorescence quantitative polymerase chain reaction was employed to determine the mRNA levels of OXA， OX1R， and OX2R in the hypothalamus. ResultCompared with the blank group， the model group had decreased body weight （P<0.01）， increased escape latency （P<0.01）， increased sleep latency （P<0.01）， shortened sleep duration （P<0.01）， elevated OXA level and lowered 5-HT level in the hypothalamus， serum， and spleen （P<0.05）， and up-regulated mRNA levels of OXA， OX1R， and OX2R in the hypothalamus （P<0.01）. Compared with the model group， the low- and high-dose groups of Shugan Jianpi Yangxin prescription showed increased body weight （P<0.05， P<0.01）， shortened escape latency （P<0.05）， shortened sleep latency and prolonged sleep duration （P<0.01）， and lowered OXA level and elevated 5-HT level in the hypothalamus， serum， and spleen （P<0.05， P<0.01）. Moreover， the two doses of Shugan Jianpi Yangxin prescription down-regulated the mRNA levels of OXA， OX1R， and OX2R in the hypothalamus （P<0.01）. ConclusionShugan Jianpi Yangxin prescription exerts sedative and hypnotic effects in mice by increasing the content of 5-HT in the brain and inhibiting the expression of OXA and its receptors in the hypothalamus.

Objective To investigate the genetic association between nonalcoholic fatty liver disease(NAFLD)and type 2 diabetes mellitus(T2DM)using bidirectional two-sample Mendelian randomization(MR),as well as the causal relationship between NAFLD and T2DM across different body mass index(BMI)categories.Methods The data were derived from genome-wide association studies conducted in European populations,with a sample size of 32 941 cases for NAFLD,312 646 cases for T2DM,and 681 275 cases for BMI.The univariate and multivariate MR methods were used to assess the bidirectional causal relationship between NAFLD and T2DM in the general population and across different BMI subtypes.The methods of inverse-variance weighting,MR-Egger regression,constrained maximum likelihood and model averaging,and weighted median were used to conduct the MR analysis,and MR-Pleiotropy Residual Sum and Outlier,radial MR,the MR-Egger intercept method,and the Cochrane Q test were used for sensitivity analysis.Results The univariate MR analysis revealed a bidirectional causal relationship between NAFLD and T2DM in the general population(forward analysis:odds ratio[OR]=9.75,95%confidence interval[CI]:2.57-37.00,P＜0.001;reverse analysis:OR=1.01,95%CI:1.00-1.01,P＜0.01).After adjustment for BMI,the multivariate MR analysis showed that the causal relationship between NAFLD and T2DM remained significant in the general population(OR=33.12,95%CI:7.57-144.95,P＜0.000 1).The subgroup analysis showed a causal relationship between NAFLD and T2DM across all BMI subtypes(lean subgroup:OR=12.19,95%CI:3.35-44.40,P＜0.001;overweight subgroup:OR=4.30,95%CI:1.69-10.92,P＜0.01;obese subgroup:OR=1.67,95%CI:1.14-2.44,P＜0.01).Conclusion This study reveals the causal relationship between NAFLD and T2DM in the general population of NAFLD and across different BMI subtypes from a genetic perspective.

OBJECTIVE To study the absorption characteristics of phenylpropanoids of Mongolian medicine Tabson-2 decoction(TBD) in Caco-2 cells and to preliminarily clarify the oral absorption mechanism of TBD. METHODS Caco-2 cell monolayer model was used to analyze the uptake components of TBD in Caco-2 cells by UPLC-MS/MS, and UPLC-MS/MS analysis method was established to determine the nine best absorbed components of TBD, protocatechuic acid, neochlorogenic acid, chlorogenic acid, cryptogenic acid, 1,5-dicaffeinate quinic acid, isochlorogenic acid C, caffeic acid, dihydrocaffeic acid, chlorogenic acid. The effects of time, concentration and P-glycoprotein inhibitor on the absorption of each component were investigated. RESULTS The overall intake of caffeic acid and dihydrocaffeic acid showed an upward trend in 0-180 min, and did not show saturation. The absorption of 3-hydroxycinnamic acid was constant at about 90 min and tended to saturation. The intakes of cryptochlorogenic acid, 1,5-dicaffeinate, quinic acid, isochlorogenic acid C, neochlorogenic acid, chlorogenic acid and protocatechuic acid first decreased and then increased with time from about 90 min. The addition of P-glycoprotein inhibitor verapamil and cyclosporin A had an effect on the absorption of dihydrocaffeic acid compared with the phenylpropanoid components, indicated that dihydrocaffeic acid was the substrate of P-glycoprotein. CONCLUSION The main phenylpropanoids of TBD enter Caco-2 mainly by passive diffusion, supplemented by active transport, and the absorption process of the other eight components is not affected by the efflux of P-glycoprotein except dihydrocaffeic acid.

Objective: To investigate characteristics of intestinal flora in elderly patients with coronary heart disease (CHD) versus CHD plus complicated heart failure (HF). Methods: The 80 CHD patients admitted into our hospital from April 2017 to April 2018 were consecutively enrolled in this prospective study.They were divided into the CHD group (without HF, n=40) and the HF group (CHD plus HF, n=40), and 40 healthy elderly people taking health examination during the same period were considered as the control group.The 0.5 g stool specimen from each observer was collected.The specific DNA fragments of 16S rDNA/18S rDNA/ITS/ function genes were amplified, the Polymerase Chain reaction(PCR)amplified products were sequenced by Illumina Genome Analyzer IIx.And the sequencing results were followed by Read splicing, OTU clustering, alpha diversity analysis and species diversity analysis.Finally, the sample species information was obtained. Results: The mean abundances of gut microflora in CHD group and in HF group were lower than in control group (P<0.05). The abundance of phylum bacteroidetes was higher in the control group (66.41±3.25)% than in the CHD group (45.21±9.07)% and in HF groups (27.29±13.27)%(P<0.05). And the abundance of phylum firmicutes was lower in the control group than in the CHD and HF groups (P<0.05). LEfSe analysis showed that the flora with high abundances in the healthy elderly group were mainly Bacteroides, Bacteroidia and Bacteroidales.Ruminococcus and Clostridium occurred mainly in the CHD group, and Firmicutes and Enterobacte occurred mainly in CHD plus heart failure group. Conclusions The intestinal flora disturbance occurs in elderly patients with coronary heart disease or CHD with heart failure.

Objective: To investigate the clinical significance and expression of microRNA(miR)-561 in esophageal squamous cell carcinoma. Methods: From January 2015 to February 2016, 96 specimens of adjacent tissues and cancer tissues from patients with esophageal squamous cell carcinoma who underwent surgery in Dongfeng Hospital Affiliated of Hubei University of Medicine were selected to detect the expression level of miR-561. Cell culture experiments were used to detect the expression level of miR-561 in Het-1a, Kyse150 and Eca109 cell lines. The correlation between clinical features and the expression level of miR-561 was counted. The expression level of miR-561 in patients with esophageal squamous cell carcinoma was analyzed. The relationship between the level of miR-561 and prognosis was investigated. Results: Relative expression of miR-561 in esophageal squamous cell carcinoma cell lines Eca109 and Kyse150 (1.61 ± 0.30, 1.21 ± 0.28) was significantly lower than that in Het-1a (2.56 ± 0.51), and the difference was statistically significant (P < 0.05). The relative expression of miR-561 in esophageal squamous cell carcinoma tissues was significantly lower than that in adjacent tissues (0.80 ± 0.17 vs. 1.51 ± 0.42), and the difference was statistically significant (P < 0.05). The expression of miR-561 in esophageal squamous cell carcinoma was correlated with alcohol drinking history, lymph node metastasis, differentiation degree and tumor stage (P < 0.05). Cox multivariate regression analysis showed that the independent risk factors affecting the prognosis of patients were the expression of miR-561, TNM stage and lymph node metastasis (P < 0.05). The survival rate of patients with esophageal squamous cell carcinoma whose expression of miR-561 did not decrease at follow-up was significantly higher than that of patients with decreased expression of miR-561 (P < 0.05). Conclusions In patients with esophageal squamous cell carcinoma, the expression of miR-561 is low, and the development, occurrence and prognosis of esophageal squamous cell carcinoma are closely related with it.

Objective The changes of Mg2+ concentration and cell apoptosis were detected by using siRNA to silence MagT1 in rat cardio myocytes.Methods MagT1 siRNA sequences were designed and synthetized,then transfected into primary cultured rat myocardial cell for silencing MagT1.The expression of MagT1 mRNA and protein were detected by RT-PCR and Western blot.The changes of Mg2+ concentration in the cells were detected by fluorescence microscopy.Flow cytometry (FCM) was used to detect the cell apoptosis.Results Compared with negative siRNA group,MagT1 siRNA transfected rat cardiomyocytes after 48 h,MagT1 mRNA silence efficiency was 51.83 ％ (P＜0.05),the silence of MagT1 protein efficiency was 56.75 ％ (P＜0.05),intracellular Mg2+ concentration was reduced by 29.13％ (P＜0.05),the apoptosis rate was 31.18％ (P＜0.01);MagT1 siRNA transfected rat cardiomyocytes after 60 h,MagT1 mRNA silence efficiency was 86.91％ (P＜0.01),the silence of MagT1 protein efficiency was 83.85％ (P＜0.01),intracellular Mg2+ concentration was reduced by 41.32％ (P＜0.01),the apoptosis rate was 40.61％ (P＜0.01).Conclusion After the silencing of MagT1,the concentration of Mg2+ in the cells decreased significantly,the apoptotic rate increased significantly,cell life activities are greatly affected.

Objective To prepare polyethylene glycol/cyclic asparagines-glycine-arginine functionalized gold nanoparticles (GNPs-PEG@cNGR) and evaluate their effectiveness in CT imaging of breast cancer angiogenesis.Methods The GNPs were synthesized by one-step reduction of chloroauric acid by sodium citrate.The thiolated PEG and cysteine-modified cNGR were coupled to the surface of GNPs through Au-S bonds,respectively.The GNPs-PEG@cNGR was characterized by transmission electron microscopy,Zeta potential/hydration particle size analyzer,and Fourier transform infrared spectrometer.The uptake and CT imaging effect of GNPs-PEG@cNGR were assessed by human umbilical vein endothelial cells (HUVEC) and human hepatoma cells (HepG2) positively expressed for aminopeptidase N (APN/CD 13).The in vivo CT imaging effects on tumor angiogenesis and biocompatibility in mice of GNPs-PEG@cNGR were studied by BALB/c mouse model of 4T1 breast cancer.Results A specific CT molecular probe,i.e.GNPs-PEG@cNGR,was successfully constructed,which can target angiogenesis.The probe was spherical,with a hydration particle size of (35.7± 1.0) nm and a Zeta potential of (-13.54± 1.12) mV,and had good stability and biocompatibility.The GNPs-PEG@cNGR has good CT imaging results and can specifically target CD13-positive HUVEC and HepG2 cells.The CT imaging results in 4T1 breast cancer mice indicated that GNPs-PEG@cNGR could be specifically enriched in the tumor tissue after injection.The CT value of tumors in GNPs-PEG@cNGRz group was higher than that of GNPs-PEG group,and the difference was statistically significant (P＜0.05).Conclusions GNPs-PEG@cNGR can specifically target CD13 positive cells and can be used as a CT contrast agent for imaging tumor angiogenesis.

A fitting method of calculating local helix parameters of proteins based on dual quaternions registration fitting (DQRFit) is proposed in this paper. First, the C and N atom coordinates of each residue in the protein structure data are extracted. Then the unregistered data and reference data are constructed using the sliding windows. The square sum of the distance of the data points before and after registration is regarded as an optimization goal. We calculate the optimal rotation matrix and the translation vector using the dual quaternion registration algorithm, and get the helix parameters of the secondary structure which contain the number of residues per turn( ), helix radius( )and helix pitch( ). Furthermore, we can achieve the fitting of three-helix parameters of , , simultaneously with the dual quaternion registration, and can adjust the sliding windows to adapt to different error levels. Compared with the traditional helix fitting method, DQRFit has some advantages such as low computational complexity, strong anti-interference, and high fitting accuracy. It is proven that the precision of proposed DQRFit for α helix detection is comparable to that of the dictionary of secondary structure of proteins (DSSP), and is better than that of other traditional methods. This is of great significance for the protein structure classification and functional prediction, drug design, protein structure visualization and other fields in the future.

Objective To observe the clinical effect of Xiaoke-Yuzu decoction on diabetic peripheral neuropathy (DPN). Methods A total of 100 DPN inpatients were recruited and randomly divided into the treatment and control groups. The two groups were both received basic therapy, while the treatment group additionally received Xiaoke-Yuzu decoction. Toronto clinical scores and Chinese medicine symptom scores of both groups were collected to evaluate the clinical effect before and after the therapy. Results The Toronto scores of treatment group were significantly lower than control group after treatment (symptoms score 1.50 ± 0.94 vs. 2.23 ± 1.01, reflection score 3.60 ± 1.77 vs. 4.27 ± 1.72, feeling test score 1.53 ± 0.63 vs. 2.10 ± 0.84,all P<0.05). Meanwhile, the Chinese medicine symptom scores of treatment group were also significantly lower than the control group (main symptom score 1.77 ± 1.17 vs. 3.17 ± 1.82, posterior symptom score 2.23 ± 1.59 vs. 4.27 ± 1.57, the tongue and pulse score 1.83 ± 0.65 vs. 2.47 ± 0.51, all P<0.05). Conclusion Xiaoke-Yuzu decoction plus basic therpy could improve the clinical symptoms of DPN patients.

Objective To develop a fluorescence signal activatable multifunctional molecular probe with theranostics function through combining ultrasmall gold nanorods(UGNRs) with fluorescein,and to evaluate its therapeutic effect on photothermal therapy (PTT) in breast cancer cells.Methods The UGNRs were synthesized by the one-pot seedless method,then the functionalized modification of UGNRs were conducted using cysteamine.Finally,the activatable ultrasmall gold nanorods (AUGNRs) were synthesized by amide condensation of —NH2 of cysteamine and —COOH of carboxylated fluorescein CyS.The cell uptake ability and GSH-mediated imaging ability of AUGNRs were studied using breast cancer 4T1 cells.4T1 cells co-cultured with AUGNRs were irradiated with 808 nm excitation light,and the PTT effects were assessed by MTT colorimetric staining and calcein-AM/PI staining.Results The AUGNRs were synthesized successfully,which could be uptaken by 4T1 cells quickly and efficiently,and could achieve intracellular glutathione (GSH) triggered fluorescence recovery.No obvious cytotoxicity of AUGNRs to 4T1 cells was observed in the co-cultivation.Moreover,obvious PTT effects could be induced by 808 nm laser,which could effectively kill 4T1 cancer cells.Conclusion The fluorescence signals of AUGNRs can be induced by intracellular GSH,and tumor cell destruction can be achieved by 808 laser-excitated PTT effects.