ObjectiveTo investigate the expression levels of miR-128-3p， SIRT1， and AMPK in the peripheral blood of patients with type 2 diabetes mellitus （T2DM） comorbid with nonalcoholic fatty liver disease （NAFLD）， as well as the role of miR-128-3p in predicting NAFLD in T2DM patients. MethodsA total of 80 patients with T2DM who were hospitalized in The First Affiliated Hospital of Anhui University of Chinese Medicine from September 2022 to August 2023 were enrolled and divided into T2DM group with 40 patients and NAFLD group with 40 patients， and according to the NAFLD fibrosis score （NFS）， the patients were further divided into progressive liver fibrosis group with 16 patients and non-progressive liver fibrosis group with 64 patients. General data and biochemical parameters were collected； quantitative real-time PCR was used to measure the mRNA expression levels of miR-128-3p， SIRT1， and AMPK in peripheral blood， and Western blot was used to measure the protein expression levels of SIRT1 and AMPK. The independent-samples t test was used for comparison of normally distributed data between two groups， and the Mann-Whitney U test was used for comparison of data with skewed distribution between two groups； the chi-square test was used for comparison of categorical data between two groups. The logistic regression analysis was used to identify the influencing factors for the presence of NAFLD and progressive liver fibrosis， and the receiver operating characteristic （ROC） curve analysis was used to determine the optimal cut-off value of miR-128-3p for predicting NAFLD. ResultsThere were significant differences between the NAFLD group and the non-NAFLD group in body mass index， fasting plasma glucose， glycated hemoglobin， fasting insulin， fasting C-peptide， alanine aminotransferase （ALT）， aspartate aminotransferase， gamma-glutamyl transpeptidase， alkaline phosphatase， fibronectin， triglycerides， high-density lipoprotein cholesterol， total triiodothyronine （TT3）， Homeostasis Model Assessment of Insulin Resistance （HOMA-IR）， and NFS （all P<0.05）. Compared with the non-NAFLD group， the NAFLD group had a significantly higher mRNA expression level of miR-128-3p in peripheral blood （t=-8.765， P<0.001） and significant reductions in the mRNA and proteins expression levels of SIRT1 and AMPK （P<0.001）. There were significant differences between the progressive liver fibrosis group and the non-progressive liver fibrosis group in age， ALT， free triiodothyronine， TT3， superoxide dismutase， and miR-128-3p （all P<0.05）. The logistic regression analysis showed that miR-128-3p was an independent risk factor for the development of NAFLD （odds ratio ［OR］=8.221， 95% confidence interval ［CI］： 2.735 — 24.714， P<0.001） and progressive liver fibrosis （OR=1.493， 95%CI： 1.117‍ ‍—‍ ‍1.997， P=0.007）. The ROC curve analysis showed that miR-128-3p had an area under the ROC curve of 0.890 （95%CI： 0.829 — 0.950）， with an optimal cut-off value of 13.165， a sensitivity of 89.3%， and a specificity of 72.7%. ConclusionThere is an increase in the expression of miR-128-3p in peripheral blood of T2DM patients with NAFLD， while there are reductions in the expression levels of SIRT1 and AMPK， suggesting that miR-128-3p has a certain diagnostic value in identifying NAFLD and liver fibrosis in such population.

Sha （痧） is an acute infectious disease characterised by the appearance of rashes on the skin， caused by exposure to epidemic toxin and pestilent qi. Sha Zhang Yu Heng （《痧胀玉衡》） discussed the treatment principles and methods， and listed collateral bloodletting as one of the main treatments. Through organizing the articles and proved cases， we found that the author believes Sha （痧） is caused by epidemic pathogen， belonging to heat toxin with rapid changes， so timely treatment for qi and blood simultaneously could achieve the effect of transforming qi into defensive qi. Sha Zhang Yu Heng focuses on patient's position during treatmet， the material of the needle， the site of treatment， the quantum of stimulation and the operation of the contraindications and other essentials. According to the depth of the disease location， use traditional Chinese herbal medicine， scraping together to identify the root of the disease. In addition， diet suggestions for the prevention of the recrudescence of disease are also described in detail.

ObjectiveTo explore the beliefs and attitudes about sleep in patients with comorbid depressive disorder and insomnia， and to explore its influence on sleep quality. MethodsPatients with comorbid depressive disorder and insomnia （n=61） and patients with primary insomnia （n=62） who met criteria specified in the Diagnostic and Statistical Manual of Mental Disorders， fourth edition （DSM-IV） in Beijing Anding Hospital Affiliated to Capital Medical University were enrolled， meantime， another 64 healthy controls were recruited. All subjects were assessed using Dysfunctional Beliefs and Attitudes about Sleep （DBAS） and Pittsburgh Sleep Quality Index （PSQI）. Additionally， patients with comorbid depressive disorder and insomnia were evaluated using Hamilton Depression Scale-17 item （HAMD-17）. The PSQI and DBAS scores were compared among three groups using analysis of covariance， and multiple linear regression analysis was used to screen the factors affecting PSQI score in patients with comorbid depressive disorder and insomnia. ResultsCompared with healthy controls， higher scores of PSQI （t=18.932， 18.610， P<0.01） along with lower scores of DBAS （t=-5.561， -5.791， P<0.01） were observed in patients with comorbid depressive disorder and insomnia and patients with primary insomnia. Taking the PSQI score of patients with comorbid depressive disorder and insomnia as the dependent variable， statistically significant equations were generated using multiple linear regression analysis （F=14.095， R²=0.327， P<0.05）， and the predictive and control factors of sleep in DBAS and age were found to be the influencing factors of PSQI score in patients （B=-0.100， -0.279， P<0.05 or 0.01）. ConclusionCompared with the normal，depression patients with insomnia have more dysfunctional beliefs and attitudes towards sleep，and dysfunctional cognition may be the influencing factor of their sleep quality.

Objective To investigate the effect of long non-coding RNA (lncRNA) LNC01309 on the proliferation and migration abilities of human hepatocellular carcinoma (HCC) cells and its mechanism of action. Methods HCC samples and corresponding adjacent tissue samples were collected from 12 patients with HCC who underwent surgical treatment in The Sixth Medical Center of PLA General Hospital from February 2018 to June 2019, and quantitative real-time PCR was used to measure the relative expression level of LNC01309. Quantitative real-time PCR was also used to measure the expression level of LNC01309 in human hepatoma cell lines (HepG2, SNU-398, and Hep3B) and the human immortalized normal liver cell line THLE-2. After LNC01309 was overexpressed in HepG2 cells, the cells were divided into plasmid control group (pEXP-control) and overexpression group (pEXP-LNC01309). CCK-8 assay was used to observe the change in cell proliferation, and wound healing assay and Transwell assay were used to observe migration ability. RNA co-immunoprecipitation was used to detect the interaction between LNC01309 with RBM38, with cells divided into IgG group and RBM38 antibody group, and cycloheximide chase assay was used to analyze the effect of LNC01309 on the stability of RBM38 protein. RBM38 was overexpressed in HepG2 cells to conduct the recovery experiment, and CCK-8 assay, wound healing assay, and Transwell assay were used to observe the changes in cell proliferation and migration abilities. The t -test was used for comparison of continuous data between two groups. Results The mean expression level of LNC01309 in HCC tissue was significantly higher than that in adjacent tissue (4.225±2.285 vs 1.541±0.530, t =3.618, P =0.004), and the relative expression level of LNC01309 in hepatoma cells (HepG2, SNU-398, and Hep3B) was also significantly higher than that in normal hepatocytes (THLE-2) ( t =4.231、6.489、14.480, all P < 0.05). Compared with the control group, HepG2 cells with the overexpression of LNC01309 had significant increases in growth rate (OD 450 value at 96 hours: 1.885±0.107 vs 2.527±0.234, t =4.330, P =0.012) and migration ability (11.65%±2.40% vs 35.66%±4.90%, t =9.837, P < 0.001; 100.00%±3.11% vs 161.00%±35.93%, t =4.399, P =0.005); however, the upregulated proliferation and migration abilities of hepatoma cells induced by LNC01309 overexpression were partially inhibited by RBM38 (OD 450 value at 96 hours: 2.500±0.227 vs 1.913±0.282, t =2.812, P =0.048; 168.00%±9.43% vs 117.20%±18.03%, t =6.622, P < 0.001). Compared with the IgG control group, RBM38 antibody significantly enriched the precipitation of LNC01309 ( t =3.846, P =0.031). The results of cycloheximide chase assay showed that the LNC01309 overexpression group had a significant reduction in the stability of RBM38 protein ( t =8.038, P =0.001). Conclusion The newly identified LNC01309 reduces the stability of RBM38 protein through interaction with RBM38 and promotes the proliferation and migration of HCC cells.

BACKGROUND: Flubendazole is an anthelmintic and categorized in benzimidazole. Previous evidence indicates its suppression on proliferation of colon cancer and breast cancer cells. Our study aims to explore the effects of flubendazole on non-small cell lung cancer A549 and H460 cell lines and the underlying mechanism. METHODS: CCK-8 assay was used to detect the effect of flubendazole at different concentrations on viability of both cell lines A549 and H460. We used western blot to detect the expression levels of autophagy-related proteins p62 and LC3 after flubendazole treatment. Cells were transfected with tandem fluorescent adenovirus (mRFP-GFP-LC3), and the impact of flubendazole treatment on autophagic flux were analyzed. RESULTS: Cell viability analysis showed a dose-dependent inhibitory effect on proliferation of both A549 and H460, comparing to cells without flubendazole treating (P<0.001). Level of p62 decreased and LC3 II/I ratio increased in cells treated with 2 μmol/L flubendazole for 24 h and 48 h, compared to control groups (P<0.005). Red fluorescence signals increased in mRFP-GFP-LC3 transfected cells after flubendazole treating, suggesting an elevation in autophagic flux. CONCLUSIONS Flubendazole may inhibit the proliferation of A549 and H460 cells and promote autophagy.

Objective To prepare biofilm scaffolds by using mesenteric acellular matrix, so as to investigate their physicochemical and biological characteristics. Methods The mesenteric tissues were subjected to trypsin digestion, and the mesenteric cells were removed after repeated freezing and thawing of mesenteric tissue. Mesenteries were divided into mesenteric matrix group (Group A) and acellular mesenteric matrix group (Group B). The physical and chemical properties of mesenteric matrix in two groups were tested by HE staining, electron microscopy, DNA detection, cytotoxicity test and tensile mechanics test. The blood flow of the vessels was detected by ultrasonography at 1st week, 1st month and 2nd month, and the vessels were observed by pathological examination. Results HE staining and electron microscopy showed that the mesentery of Group B was loose in acellular mesentery matrix, and the arrangement of fibers was neat, with no cells remaining. Compared with Group A, the expression level of DNA in Group B was lower, with more completely decellularized cells. CCK-8 cytotoxicity test showed that there was no cytotoxicity in Group A and Group B. FDA-PI fluorescence staining showed no cytotoxicity of cells in both groups. Cells in Croup A and Group B survived well, and no dead cells were found. Tensile mechanics test showed that there were no significant differences in maximum tensile force, maximum elongation, yield strength, yield point elongation between Group A and Group B. The early patency of acellular mesenteric stent implantation was good, and endothelial hyperplasia was obvious at 2nd month after stent implantation. Conclusion sMesenteric cells were removed by freeze-thaw and enzymatic digestion. Mesenteric stroma was completely removed without cytotoxicity, which showed good mechanical characteristics. Mesenteric stent implantation had good early patency and endothelial proliferation after 2 months.

Objective To construct a model of small caliber vascular endothelial growth factor(VEGF) in tissue engineering,to investigate the performance of the sustained-release microspheres and vascular stent,and to provide materials and theoretical basis for animal experiment.Methods The sheep carotid arteries were treated with a cellular reagents,the cellular conditions and the stent properties were observed.Preparation of sustained release microspheres containing VEGF,particle size,encapsulation efficiency,drug loading and release curve were measured.The effective combination of the slow release microsphere and the vascular stent was used in the freeze drying technology.The rat vascular endothelial cells grown in tissue engineered blood vessel model release lumen,observe the growth of endothelial cells.Results After the treatment,the original performance of the vascular stent can be maintained.The average particle size of the microspheres was (9.8 ± 6.0) μm,which could be released slowly in 20 days,and the release rate was 70％.Microspheres can effectively with the tissue.engineering blood vessel tight binding.Rat vascular endothelial cells can grow in the vascular stent surface.Conclusion Using Triton X-100,DNA/RNA ribozyme for acellular reagent,stent performance is good.PLGA microspheres have good sustained release performance,and constructing appropriate tissue engineered small caliber vascular release model by using freeze drying technology can make the stent compact structure.

Objective:To conduct a systematic study of the immunologic response of rats to transplanted glutaraldehyde ( GA)-treated porcine blood vessels in vivo.Methods: The experiment was divided into two groups:fresh group and glutaraldehyde-treated group.Twenty cases of fresh and glutaraldehyde-treated porcine pulmonary arteries were subcutaneously embedded in rats.We compared the changes using HE staining and immunohistochemistry.Results:HE staining showed that there were stronger expression on day 12 and day 30 in the fresh group than that in the glutaraldehyde group.There were similar results in morphology in CD68,C3,IgG.The results of integral optical density ( IOD) in immunohistochemistry showed that IOD started rising from day 4 and got the peak on day 12 or day 30 and or fell on day 60.Conclusion: Innate immunity played an important role in the research on xenogenic immunological rejection mechanism.The immunogenicity of glutaraldehyde-treated xenogenic blood vessels is lower than that in fresh blood vessels.However there is still immunogenicity in glutaraldehyde-treated xenogenic blood vessels.We will explore better ways to obviously weaken the rejection.

Objective:To observe serum cystatin C (Cys C)level in patients before and after off-pump coronary ar-tery bypass grafting (OPCABG),and explore its relationship with coronary artery lesion.Methods:A total of 321 patients,who received OPCABG in department of heart surgery of our hospital from Aug 2013 to Nov 2013,were selected.According to number of diseased coronary vessels,they were divided into single-vessel group (n= 11), double-vessel group (n=33)and multi-vessel group (n=277).Double antibody enzyme linked immunosorbent assay (ELISA)was used to measure serum Cys C level in all groups.Results:The Cys C levels in single-vessel group, double-vessel group,multi-vessel group before OPCABG were (0.732 ± 0.286)mg/L, (0.807 ± 0.265)mg/L, (0.911±0.273)mg/L respectively,there were significant difference (F=3.942,P =0.038);and after OPCABG were (0.616±0.198)mg/L,(0.607±0.201)mg/L, (0.646±0.166)mg/L respectively ,there were no signifi-cant difference (F=0.493,P =0.617),namely there was no significant relationship between coronary artery disease extent and Cys C level after OPCABG,P >0.05. Conclusion:There is no significant relationship between original coronary artery disease extent and serum cystatin C level after off-pump coronary artery bypass grafting,namely off-pump coronary artery bypass grafting is an effective therapeutic measure for patients with coronary heart disease.

Objective To explore the event-related potentials (ERPs) P300 is changeable or not before and after treatment. Methods 99 cases of patients with first onset of depression diagnosed by DSM-Ⅳ as case group,and 100 cases matched with patients as control group were collected. P300 of two groups were obtained before and after treatment for 6 weeks,12 weeks,24 weeks. T test was used to analysis the difference of indicators of P300 among groups; repeated measure analysis of variance was used to analysis the longitudinal changes. Results shorter latency of N2-P3 ( (P ＜ 0.01 ); and lower amplitude of N2, P3, N2-P3 (P ＜ 0. 05 ), higher amplitude of P2-tency and a upward one in N2-P3 latency in the four periods; a upward trend could also be found in P3, N2-P3 amplitude, but there were no statistical differences(P ＞ 0. 05 ). The results of paired-samples t test: P3, N2-P3 amplitude in case group were higher after treatment for 6 weeks than before, the difference was significant (P ＜ 0.01 ); no significant results were found in P300 latency or amplitude between the 62 cases of depression after treatment for 24 weeks and the 65 normal controls selected (P ＞ 0. 05 ). Conclusion P300 latencies and amplitudes tend to be partly recovered after the acute treatment in patients with depression, but after the long-term therapy not clear.