Adult ; Female ; Humans ; Liver ; pathology ; Male ; Middle Aged ; Non-alcoholic Fatty Liver Disease ; pathology ; Obesity ; pathology ; Young Adult

Objective To evaluate the prevalence of hepatic steatosis in patients with chronic hepatitis B (CHB) and the impact of hepatic steatosis on the progress of fibrosis. Methods Five hundred and sixty two untreated CHB patients (405 males and 157 females) with an average age of 31.3 underwent liver biopsy from January to August 2007. On the day of liver biopsy, a questionnaire was completed and a blood sample was obtained for laboratory analysis. The degree of liver steatosis, necroinflammation and fibrosis was assessed; demographic information and clinical data including age, gender, body mass index (BMI), history of diabetes mellitus, hypertension, dyslipidemia, and HBeAg status, HBV DNA viral load were documented. Results In 562 patients, 102 (18. 2% ) had steatosis. Multivariate analysis demonstrated that liver steatosis was associated with the levels of TG, APO-B, UA, FSG, and higher BMI; and the progress of fibrosis was associated with high degree of hepatic steatosis and necroinflammation, age over 35 years, HBV DNA > 103 copies/L, high BMI and GGT. Conclusions The results show that obesity and dyslipidemia in CHB patients are associated with the hepatic steatosis, and the latter seems to be an important determinant for fibrosis.

Objective: To investigate the clinical features and reliable diagnostic method in HIV/AIDS patients with smear negative pulmonary tuberculosis (TB). Methods: Clinical data of 112 HIV/AIDS patients complicated with smear negative pulmonary TB who were treated in our hospital from January 2013 to September 2015 were retrospectively analyzed. These clinical data includeded clinical symptom, blood routine test, blood biochemistry, T lymphocyte subsets classification, sputum acid-fast bacillus smear, mycobacterium tuberculosis culture, purified protein derivatives tuberculin (PPD) test, interferon gamma-release assay for Mycobacterium tuberculosis (T-SPOT.TB), TB-DNA and chest computed tomography (CT). Diagnostic specificity and sensitivity of these parameters were analyzed. Results: No specific clinical manifestation of these patients was identified. The chest CT feature was also atypical. The positive rates including T-SPOT, TB, TB-DNA and PPD test were all low. The positive rates of T-SPOT.TB and PPD test in patients with a CD4+ cell count >200 cells/μl was significantly higher than that of patients with a CD4+ cell count ≤50 cells/μl and 51≤CD4≤200 cells/μl (P<0.01). Conclusions The clinical feature of smear negative pulmonary TB in HIV/AIDS patients is atypical. For a definite diagnosis, a comprehensive analysis with clinical manifestations, laboratory test, imaging examination and etiologic detection is required.

Lumbosacral nerve root anomalies (LNRA) refers to the abnormality of lumbosacral nerve root in morphology, course and initial position, which may be related to the abnormal migration of nerve root in embryo. Imaging examination is a commonly used method for localization and qualitative diagnosis of LNRA. Clinically, many classification systems have been established based on imaging results, autopsy reports and Book observations. However, the lack of evaluation on the advantages and disadvantages of each classification is not conducive to the development of clinical work. In this paper, the classification and characteristics of LNRA at home and abroad were reviewed, and the advantages and limitations of each classification were evaluated. Through the literature retrieval and summary, it is not reliable to design the corresponding classification based on the imaging results. Not only is the detection rate of MRI as the preferred examination method far lower than that of autopsy, but also it is not an intuitive study of the lesions, and its authenticity is questionable. Based on the classification of autopsy results, the spinal cord is directly stripped in the study, excluding the surrounding soft tissue and bone marrow. The interference of spinal bone structure is more conducive to the comprehensive observation of variation, which has a certain guiding significance for clinical; based on the classification of intraoperative observation results, the number of research samples is large, easy to obtain, and there are matching imaging data to do the follow-up comparative study, among which the Neidre classification is themost widely used classification, which provides considerable convenience for clinical work. LNRA may not be treated as a common disease due to the limitation of technology and lack of understanding. The lack of specific symptoms and the characteristics of combined compression will affect the detection rate. In terms of treatment, traditional discectomy and spinal canal decompression cannot meet the clinical needs. More new surgical methods need to be proposed to improve the prognosis and quality of life of patients.

Digital loop-mediated isothermal amplification (digital LAMP, dLAMP) is a novel nucleic acid amplification technique developed in recent years. It divides the target nucleic acid and LAMP reagent into a large number of independent detection regions, and uses a highly active chain replacement DNA polymerase and four specially designed primers for rapid amplification under isothermal conditions, which provides a good platform for quantitative detection of target nucleic acids. The advantages of high accuracy, high sensitivity, absolute quantification, high tolerance to inhibitors and simple instrumentation make the dLAMP technique very promising in molecular diagnosis, especially in rapid detection of pathogenic microorganisms, it shows a good application prospect in the fields of clinical diagnosis, food safety and environmental monitoring. Certainly, the development of dLAMP still faces some challenges, such as how to avoid non-specific amplification in multiple primer designs, multi-target nucleic acids and simultaneous detection of a large number of samples. With the development of dLAMP technology, this technology will greatly enrich the future development of molecular diagnostics. Applying rapid and effective molecular diagnostic techniques to the diagnosis of pathogenic microorganisms has important social significance for disease prevention and control.

Digital loop-mediated isothermal amplification (digital LAMP, dLAMP) is a novel nucleic acid amplification technique developed in recent years. It divides the target nucleic acid and LAMP reagent into a large number of independent detection regions, and uses a highly active chain replacement DNA polymerase and four specially designed primers for rapid amplification under isothermal conditions, which provides a good platform for quantitative detection of target nucleic acids. The advantages of high accuracy, high sensitivity, absolute quantification, high tolerance to inhibitors and simple instrumentation make the dLAMP technique very promising in molecular diagnosis, especially in rapid detection of pathogenic microorganisms, it shows a good application prospect in the fields of clinical diagnosis, food safety and environmental monitoring. Certainly, the development of dLAMP still faces some challenges, such as how to avoid non-specific amplification in multiple primer designs, multi-target nucleic acids and simultaneous detection of a large number of samples. With the development of dLAMP technology, this technology will greatly enrich the future development of molecular diagnostics. Applying rapid and effective molecular diagnostic techniques to the diagnosis of pathogenic microorganisms has important social significance for disease prevention and control.

OBJECTIVETo develop and evaluate a mouse model of nonalcoholic steatohepatitis (NASH) induced by a high-fat and high-fructose (HFHFr) diet.

METHODSSix-week-old C3H mice were randomly divided into groups for HFHFr diet experimental modeling, high fat-only (HF) diet controls, high fructose-only (HFr) diet controls, and standard chow (SC) diet controls. The standard HFHFr diet was modified so that it consisted of 76.5% standard chow, 12% lard, 1% cholesterol, 5% egg yolk powder, 5% whole milk powder, and 0.5% sodium cholate, along with 20% fructose drinking water. At the end of experimental weeks 4, 8, and 16, measurements were taken for the NASH-related parameters of body mass, serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST), lipid profile, and wet liver weight (upon sacrifice). In addition, histological changes in the liver were evaluated by hematoxylin-eosin (HE) and oil red O staining. The significance of differences between groups was assessed by statistical analysis, using the

METHODSof t-test, Wilcoxon rank sum test, x2 test, F test or Fisher's test as appropriate.

RESULTSAs compared to the mice in the SC group at the corresponding time points, the mice in the HFHFr and HF groups showed significantly higher body mass and wet liver weight, as well as more extensive and robust lipid disposition in hepatic tissues as evidenced by oil red O staining. However, HE staining indicated that the HFHFr and HF groups had different degrees of macrosteatosis accompanied with intralobular inflammatory foci, with the former showing more remarkable NASH-related histological changes. Analysis at the end of week 16 showed that about 80% of the mice in the HFHFr group had developed NASH [nonalcoholic fatty liver disease (NAFLD) activity score (NAS): less than 5]. The levels of low-and high-density lipoprotein (LDL and HDL) cholesterol, as well as the levels of ALT and AST, were increased from the end of week 4 to the end of week 8 for the HFHFr and HF groups. At the end of week 16, the two groups differed in the extent of increase in total cholesterol and LDL and HDL cholesterol, with only the HFHFr group showing statistically significant changes. Specifically, at the end of week 16, the HFHFr group showed ALT levels of 108.5 +/- 93.34 U/L (F=5.099, P =0.005 vs. HF group: 44.30 +/- 35.71 U/L, HFr group: 46.70 +/- 17.95 U/L, SC group: 24.70 +/- 6.57 U/L), AST levels of 316.30 +/- 208.98 U/L (F=6.654, P=0.001 vs. HF: 132.12 +/- 75.43 U/L, HFr: 143.30 +/- 38.53 U/L, SC: 122.60 +/- 12.76 U/L), total cholesterol levels of 5.18 +/- 0.58 mmol/L (F=72: 470, P =0.000 vs. HF: 3.94 +/- 0.75 mmol/L, HFr: 2.30 +/- 0.50 mmol/L, SC: 2.02 +/- 0.24 mmol/L), HDL cholesterol levels of 3.05 +/- 0.49 mmol/L (F=25.413, P =0.000 vs. HF: 2.65 +/- 0.54 mmol/L HFr: 1.77 +/- 0.47 mmol/L, SC: 1.58 +/- 0.16 mmol/L), LDL cholesterol levels of 1.11 +/- 0.23 mmol/L (F =83.297, P =0.000 vs. HF: 0.72 +/- 0.17 mmol/L, HFr: 0.27 +/- 0.04 mmol/L, SC: 0.20 +/- 0.05 mmol/ L).

CONCLUSIONThe present study suggests that a mouse model of NASH can be successfully induced by a 16-week modified HFHFr diet.

Purpose/Significance To alleviate digital health technology anxiety in elderly patients with chronic diseases.Method/Process A convenience sampling method is used to survey 1 222 elderly patients with chronic diseases in tertiary,secondary,and commu-nity hospitals in Shanghai,China,with respect to demographic information,the level of anxiety about digital health technologies,and the conditions of individual use of digital health technologies.Logistic regression analyse is used to investigate the factors influencing anxiety in the use of digital health technologies among elderly patients with chronic diseases.Result/Conclusion Elderly patients with chronic diseases as a whole have low moderate levels of anxiety about digital health technologies.Educational level,per capita monthly income,experience of using digital health technologies,and learning ability are independent factors affecting anxiety about digital health technolo-gies among elderly patients with chronic diseases.Aging-friendly design,assistance and support,improved facility accessibility,and precise training should be implemented so as to increase the acceptance and use of digital health technologies among elderly chronic dis-ease patients.

ObjectiveTo identify the members of the Rehmannia glutinosa miR166 gene family and clarify the response mode under adversity. MethodHigh-throughput sequencing technology was employed to obtain a small RNA database and the miR166 family members of R. glutinosa were screened out. The precursor structures were analyzed by RNAfold. DNAMAN and MEGA were used for conservative and evolutionary analyses， respectively. TargetFinder software was used to predict the target genes of R. glutinosa miR166 family members. The expression of miR166 family members in response to abiotic stress was analyzed by real-time polymerase chain reaction（Real-time PCR）. ResultFive miR166s were identified with precursors possessing complete stem-loop structures. As revealed by sequence alignment results， the precursors and matures were both highly conserved. Forty-eight target genes of miR166s were predicted， which were mainly annotated to the HD-ZIP Ⅲ family transcription factors. The expression characteristics showed that the expression of miR160s was up-regulated after R. glutinosa was infected by endophytic fungi， which was different from the expression of the family members under abiotic stress. The expression level of rgl-miR166b-5p in the drought-flood treatment group and the high-low temperature treatment group was significantly down-regulated compared with that in the control group， and the expression pattern was opposite under the endophytic fungal infection. ConclusionThe results of this study preliminarily clarified the expression patterns of R. glutinosa in response to biotic and abiotic stresses and provided a theoretical basis for future breeding and improvement of R. glutinosa.

Objective: To investigate the role of neutrophil elastase inhibitor, sivelestat, in preventing and treating nonalcoholic steatohepatitis (NASH) and its underling mechanisms. Methods: A total of forty 4-week-old male C57BL/6J ApoE-/-mice were equally divided into the following four groups: standard chow (SC)+isotonic saline; SC+sivelestat; high-fat, high-cholesterol (HFHC) diet+isotonic saline; and HFHC+sivelestat. These mice were treated with above methods for 12 weeks. Blood and liver tissue samples were collected to measure biochemical parameters, hepatic steatosis and non-alcoholic fatty liver disease (NAFLD) activity score (inflammation) were evaluated by oil red O staining and HE staining, respectively. The mRNA and protein expression levels of hepatic inflammatory cytokines, CD68, and F4/80 were determined by quantitative RT-PCR and immunohistochemistry, respectively. Comparison of means between the four groups was made by one-way analysis of variance, and comparison between any two groups was made by the LSD or SNK method (for data with homogeneity of variance) or the Tamhane or Dunnett method (for data with heterogeneity of variance). Results: Mice fed with an HFHC diet for 12 weeks developed typical pathological features of NASH compared with those fed with SC. Compared with mice fed with HFHC diet without sivelestat, those treated with HFHC and sivelestat exhibited the following features: (1) significantly reduced fast blood glucose, blood cholesterol, and hepatic biochemical parameters, as well as increased insulin sensitivity; (2) significantly reduced NAFLD activity score (5.71±1.11 vs 3.16±1.16, P < 0.05); (3) reduced monocyte chemoattractant protein-1 and tumor necrosis factor -α; (4) significantly reduced mRNA levels of CD68 and F4/80; and (5) reduced expression of CD68 in the liver. Conclusion Sivelestat alleviates the hepatic steatosis and inflammation of NASH in mice by inhibiting the activation of Kupffer cells.