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.

The patient was diagnosed with "motor retardation for 3 years" in the neurology department of our hospital,accompanied by static tremors and ataxia. Physical examination showed that there was a tendency to decrease for amplitude of repetitive finger movements of the left hand and the left hand clenched fist and toe tapping movements were slightly slower. The left heel knee tibia test and rotation test were not accurate,and the father and elder sister had similar symptoms. After admission,laboratory tests such as urinary tract ultrasound and residual urine in the bladder were completed,and the tests did not reveal any significant abnormalities. Brain MRI shows scattered non-specific white matter changes in the left parietal lobe. Levodopa challenge test was positive. The patient was then suspected to have Parkinsonism. considering that the patient has a family history. Genetic testing was ordered and the results showed abnormal amplification of the ATXN2 gene CAG sequence. Therefore,the final diagnosis was spinocerebellar ataxia type 2. The patient's symptoms are stable and there is no progression during the follow-up. SCA2 should be considered in the case of Parkinsonism accompanied by ataxia and a family history to avoid misdiagnosis and missed diagnosis.

ObjectiveTo evaluate the methodological and reporting quality of clinical practice guidelines for Chinese patent medicine （CPM） with internationally recognized tools the appraisal of guidelines for research and evaluation （AGEREE） Ⅱ and reporting items for practice guidelines in healthcare （RIGHT）， thereby providing refe-rence for the clinical application and future development of CPM guidelines. MethodsDatabases including CNKI， VIP， Wanfang and Sinomed were searched for CPM guidelines， as well as medlive.cn， websites of China Association of Chinese Medicine and Chinese Medical Association， and reference lists of the included papers. The quality of the guidelines was evaluated using the AGREE Ⅱand RIGHT tools， and consistency tests were performed using Interclass Correlation Coefficient， and descriptive analysis and chi-square test were used to analyze the reporting rate for each domain and the average score for each item. ResultsFinally， 140 CPM guidelines were included， of which 51 were disease-oriented and 89 were drug-oriented， all of which were issued by China. For 51 disease-oriented CPM guidelines， the highest average score of all six AGREE Ⅱ domains was 73.32% for clarity， and the lowest was 26.80% for application； for 89 drug-oriented CPM guidelines， the highest average score was 55.62% for scope and purpose， and the lowest was 31.32% for rigour of development. In terms of the seven domains of the RIGHT checklist， the highest reporting rate was 68.26% for background， and lowest was 27.45% for other areas regarding the disease-oriented CPM guidelines； the highest reporting rate was 61.31% for background， and the lowest was 4.49% for other areas regarding drug-oriented CPM guidelines. The average reporting rate was higher for disease-oriented than drug-oriented CPM guidelines in three domains of AGREE Ⅱ （rigour of development， clarity of presentation， editorial independence）， as well as four domains of RIGHT checklist （basic information， evidence， funding and declaration and management of interests， and other areas）. ConclusionThe overall methodology and reporting quality of the current CPM guidelines still need to be improved. It is recommended that future guideline development teams should strictly refer to the AGREE Ⅱ and RIGHT checklist， and take into account of the characteristics of CPM guidelines and relevant methodo-logical suggestions in the development and reporting of CPM guidelines， thereby guiding the clinical use of CPM in a better way.

Objective: To conduct a scoping review on prognostic prediction models for patients with comorbidity of chronic diseases, and understand modeling methods, predictive factors and predictive effect of the models, so as to provide the reference for prognostic evaluation on patients with comorbidity of chronic diseases. Methods: Literature on prognostic prediction models for patients with comorbidity of chronic diseases was collected through SinoMed, CNKI, Wanfang Data, VIP, PubMed, Embase, Cochrane Library and Web of Science published from the time of their establishment to November 1, 2023. The quality of literature was assessed using prediction model risk of bias assessment tool (PROBAST), then modeling methods, predictive factors and predictive effects were reviewed. Results: Totally 2 130 publications were retrieved, and nine publications were finally enrolled, with an overall high risk of bias. Thirteen models were involved, with three established using machine learning methods and ten established using logistic regression. The prediction results of four models were death, with main predictive factors being age, gender, body mass index (BMI), Barthel index and pressure ulcers; the prediction results of nine models were rehospitalization, with main predictive factors being age, BMI, hospitalization frequency, duration of hospital stay and hospitalization costs. Eleven models reported the area under the receiver operating characteristic curve (AUC), ranging from 0.663 to 0.991 6; two models reported the C-index, ranging from 0.64 to 0.70. Eight models performed internal validation, one model performed external validation, and four models did not reported verification methods. Conclusions The prognostic prediction models for patients with comorbidity of chronic diseases are established by logistic regression and machine learning methods with common nursing evaluation indicators, and perform well. Laboratory indicators should be considered to add in the models to further improve the predictive effects.

The patient was diagnosed with "motor retardation for 3 years" in the neurology department of our hospital,accompanied by static tremors and ataxia. Physical examination showed that there was a tendency to decrease for amplitude of repetitive finger movements of the left hand and the left hand clenched fist and toe tapping movements were slightly slower. The left heel knee tibia test and rotation test were not accurate,and the father and elder sister had similar symptoms. After admission,laboratory tests such as urinary tract ultrasound and residual urine in the bladder were completed,and the tests did not reveal any significant abnormalities. Brain MRI shows scattered non-specific white matter changes in the left parietal lobe. Levodopa challenge test was positive. The patient was then suspected to have Parkinsonism. considering that the patient has a family history. Genetic testing was ordered and the results showed abnormal amplification of the ATXN2 gene CAG sequence. Therefore,the final diagnosis was spinocerebellar ataxia type 2. The patient's symptoms are stable and there is no progression during the follow-up. SCA2 should be considered in the case of Parkinsonism accompanied by ataxia and a family history to avoid misdiagnosis and missed diagnosis.

Objective: To systematically evaluate risk factors for cardiometabolic multimorbidity (CMM), so as to provide the evidence for formulating CMM prevention and control strategies. Methods: Publications pertaining to the risk factors for CMM were retrieved from databases, including SinoMed, CNKI, Wanfang Data, VIP, PubMed and Cochrane Library from inception to March 31, 2023. Meta-analysis was performed using the software RevMan 5.4 and Stata 16.0, and sensitivity analysis was performed using the leave-one-out method. The publication bias was evaluated using Egger's test. Results: Totally 494 publications were screened, and 20 publications were included in the final analysis, including 13 cohort studies (covering 1 940 000 participants) and 7 cross-sectional studies (covering 13 000 000 participants). Meta-analysis revealed that female (OR=1.54, 95%CI: 1.40-1.71), middle age (OR=3.80, 95%CI: 3.33-4.34), elderly (OR=2.82, 95%CI: 1.48-5.37), urban resident (OR=1.41, 95%CI: 1.27-1.57), higher education level (OR=2.01, 95%CI: 1.35-3.01), higher economic level (OR=1.21, 95%CI: 1.16-1.25), overweight (OR=1.92, 95%CI: 1.64-2.26), obesity (OR=3.01, 95%CI: 2.30-3.93), central obesity (OR=1.70, 95%CI: 1.12-2.56), smoking (OR=1.27, 95%CI: 1.07-1.51), alcohol consumption (OR=1.27, 95%CI: 1.01-1.59), irregular diet (OR=1.10, 95%CI: 1.02-1.18), insufficient intake of vegetables and fruits (OR=1.12, 95%CI: 1.07-1.17), lack of sleep at night (OR=1.17, 95%CI: 1.08-1.27), and depression (OR=1.50, 95%CI: 1.33-1.69) were risk factors for CMM. Sensitivity analysis of effects of central obesity and alcohol consumption were not robust. No publication bias was examined by Egger's test. Conclusions Female, middle age, elderly, urban resident, higher education level, higher economic level, overweight, obesity, central obesity, smoking, alcohol consumption, irregular diet, insufficient intake of vegetables and fruits, lack of sleep at night and depression are risk factors for CMM.

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 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 study multiple-drug resistance expression of CNE1 and CNE2 cell lines induced by radiation and its reversal by cyclosporine A and interferon. Methods Drug-resistant CNE1 and CNE2 cell lines were induced by radiation, and expression of P-gp, MRP, TOPⅡ and GST-? on protein and mRNA level were analyzed by flow cytometry and reverse transcription-polymerase chain reaction methods before and after radiation. At the same time, effect of cyclosporine A and interferon on multiple-drug resistance expression induced by radiation was studied. Results The results showed that expression of P-gp, MRP and GST-? proteins on CNE1 and CNE2 cell lines were up-regulated by radiation, and cyclosporine A and interferon could reverse the up-regulation of P-gp and MRP proteins induced by radiation. Conclusion Radiation can induce multiple-drug resistance in CNE1 and CNE2 cell lines, and it can be reversed by cyclosporine A and interferon.