Objective:To establish the alcoholic fatty liver(AFL) animal models,and to explore the protective effect of Zhi Zi Da Huang Tang (ZZDHT) on the rats with AFL and its dosage.Methods:A total of 54 SD rats were divided into normal control group (n=10) and model control group (n=44).The rats in model control group were given alcohol by lavage (50% ethanol solution 6.0 mL·kg-1) combined with high-fat feed to establish the rat models of AFL.After 4 weeks,the rats in model control group were randomly divided into model group(treated with water),simvastatin group (10 mg·kg-1),low and high doses (5.0 and 10.0 g·kg-1) of ZZDHT groups,and there were 10 rats in each group.Every 2 weeks during the process,the body weights and levels of serum total cholesterol (TC),triglyceride (TG),aspertate aminotransferase (AST),and alanine aminotransferase (ALT) were detected.After 4 weeks,the body weights and liver weights of the rats were detected;the levels of TC,TG,AST,ALT,alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) in serum and liver tissue of the rats of were detected;the levels of tumor necrosis factor-alpha (TNF-α),interleukin-6 (IL-6) and interleukin-1β(IL-1β) in serum of the rats were detected;the morphology of liver tissue was observed by HE staining and the pathological examination was performed.Results:Compared with normal control group,the levels of serum TC,AST,ALT,TG,TNF-α,IL-6,and IL-1β of the rats in model group 4 weeks after administration were significantly increased (P<0.05 or P<0.01).Compared with model group,the levels of serum TC,TG,AST,and ALT of the rats in low and high doses of ZZDHT groups 4 weeks after administration were decreased (P<0.05 or P<0.01);the levels of serum TNF-α and IL-6,IL-1β of the rats in high dose of ZZDHT group were decreased (P<0.01);the levels of serum TNF-α and IL-1β of the rats in low dose of ZZDHT group were decreased (P<0.01);the levels of serum ADH and ALDH in liver tissue of the rats in low and high doses of ZZDHT groups and simvastatin group were decreased(P<0.05 or P<0.01).The HE staining result showed that compared with model group,the pathological conditions of the liver tissue of the AFL rats in ZZDHT groups were significantly improved.Conclusion:ZZDHT can significantly improve the liver injury caused by high fat diet combined with alcohol and fat liver lesions.

Objective To investigate the expression of LOXL2 protein (lysyl oxidase like-2 protein) and epithelial-mesenchymal transition (EMT) related markers in cholangiocarcinoma tissues and its relation with the malignant features. Methods The expression of LOXL2、E-cadherin and Vimentin protein in 48 cases of cholangiocarcinoma tissues was detected by immunohistochemistry and compared with the clinicopathological data of cholangiocarcinoma. Results The positive expression rate in cholangiocarcinoma was 71% ( 34/48 ) for LOXL2 and 46% ( 22/48 ) for Vimentin, the absent expression rate was 52% (25/48) for E-cadherin. The positive expression rate of LOXL2 was significantly associated with the absent expression of epithelium markers E-cadherin ( r = 0. 394, P < 0. 05 ) and the positive expression of fibroblast markers Vimentin ( r = 0. 406, P < 0. 05 ). There was no correlation between the expression of LOXL2 and patients gender, age, and cancer differentiation, but a significant correlation with tumor metastasis was found ( P < 0. 05 ). Conclusions LOXL2 protein overexpression in cholangiocarcinoma may accelerate invasion of cholangiocarcinoma through induced EMT.

Objective To investigate the correlation of the expression of KAI1/CD82 and laminin receptor (LNR) in cholangiocarcinoma, and study its role in the invasion and metastasis of cholangiocarcinoma. Methods The expressions of KAI1/CD82 and LNR in 48 cholangiocarcinoma tissue samples were detected by SP immunohistochemistry, and their relationships with clinicopathological factors were analyzed. Results The positive expression rates of KAI1/CD82 and LNR in cholangiocarcinoma were 31% (15/48) and 54% (26/48), respectively. In highly differentiated cholangiocarcinoma, the positive expression rate of KAI1/CD82 was high (χ2=3.911, P<0.05), while that of the LNR was low (χ2=6.970, P<0.05). The positive expression rate of KAI1/CD82 in cholangiocarcinoma with metastasis was significantly lower than that in cholangiocarcinoma without metastasis (χ2=5.765, P<0.05), while the positive expression rate of LNR in cholangiocarcinoma with metastasis was significantly higher than that in cholangiocarcinoma without metastasis (χ2= 9.952, P<0.05). The expression level of KAI1/CD82 was negatively correlated with that of the LNR ( r = -0.462, P < 0.01 ). Conclusions The up-regulated expression of LNR in cholangiocarcinoma correlates with the decreased expression of KAI1/CD82, and plays an important role in the invasion and metastasis of cholangio-carcinoma.

Objective To discuss the comparability of blood urea nitrogen (BUN),creatinine (Cr),uric acid (UA) results and provide basis for result concordance in different biochemical systems.Methods The within-run precision and the between-day precision for the median and high values of quality control materials were tested in Beckman LX-20 and Hitachi 7600 chemistry analyzer respectively,and the evaluation criteria were according to the precision requirement of manufacturer and our laboratory.According to American Clinical and Laboratory Standards Institute (CLSI) document EP9-A2,serum BUN,Cr,UA from 40 patients were detected by Hitachi 7600 chemistry analyzer as experimental method and by Beckman LX-20 chemistry analyzer as comparative method.The relative bias ( SE％) between experimental method and comparative method was calculated and the concordance of biochemical analysis system was determined according to EQA requirements and acceptable performance criteria Results The between-run and between-day precision coefficients of variation for BUN,Cr and UA in two biochemical analysis systems were lower than the precision requirement of manufacturer and our laboratory.In Hitachi 7600 chemistry analyzer,the SE％ for BUN at three levels of medical decision were 9.4％,2.0％ and 0.8％,the SE％ for Cr were 7.3％,3.8％ and 2.8％,the SE％ for UA were 7.9％,1.7％ and 1.2％.Two biochemical analysis systems were not comparable in BUN analysis because SE％ of serum BUN levels detected by Hitachi 7600 at low medical decision level exceeded the specified range.The remaining items between two biochemical analysis systems were concordant.Conclusion While the same test is detected by more than two systems in one laboratory,it is necessary to do method comparison and bias evaluation for concordance in order to ensure the comparability among test results.

Objective To explore the surgical management of chronic pancreatitis complicated with pancreatolithiasis (CPPL). Methods The clinical data of 66 patients with CPPL were retrospectively analyzed. Pancreatolithiasis was classified into 4 types according to the location of stones: stones located in the head of the pancreas (type Ⅰ, n=28), stones located in the body of the pancreas (type Ⅱ, n=30), stones located in the tail of the pancreas (type Ⅲ, n=1) and stones located from the head to tail of the main duct of pancreas (type Ⅳ, n=7). Ten patients (including 4 with type Ⅰpancreatolithiasis, 5 with type Ⅱ and 1 with type Ⅳ) received conservative treatment; 10 patients with type Ⅰ pancreatolithiasis underwent lithotomy under endoscope; pancreaticoduodenectomy and Beger procedure were carried out on 14 patients with type Ⅰ pancreatolithiasis, pancreatolithotomy+pancreaticojejunostomy on 25 patients with type Ⅱ pancreatolithiasis, resection of pancreatic tail and spleen on 1 patient with type Ⅲ pancreatolithiasis, and Puestow-Gillesby procedure, dividing of the neck of pancreas+removing stones from both ends of pancreatic duct+Roux-en-Y pancreatojejunostomy on 6 patients with type Ⅳ pancreatolithiasis. Results Sixty-two patients were followed up for 2 months to 15 years, and the number of patients with recurrence for type Ⅰ, Ⅱ, Ⅲ and Ⅳ pancreatolithiasis was 4, 2, 0 and 3, respectively. Conclusions Early surgical management according to the location of stones should be carried out after confirmed diagnosis of CPPL. Individualized management based on correct diagnosis and classification plays an important role in the prevention of pancreatolithiasis recurrence.

Acute leukemia marks the main type of non-solid cancer in children.Studies have proved that long non-coding RNA plays important roles in the pathogenesis of hematological malignancies and can be applied as a biomarker for diagnosis, prognosis and efficacy monitoring of leukemia patients.This article briefly reviews the roles of long non-coding RNA in the tumorigenesis and progression of childhood acute leukemia.

OBJECTIVETo explore the genetic basis and phenotypic correlation with disease severity in a large cohort of Chinese patients with hypertrophic cardiomyopathy (HCM).

METHODSA total of 179 unrelated Chinese HCM patients admitted to our department from 2002 to 2011 were enrolled in this study. Direct gene sequencing of β-myosin heavy chain (MYH7), myosin binding protein-C ( MYBPC3), and cardiac troponin T (TNNT2) were performed and clinical data were obtained in these patients.

RESULTSA total of 34 mutations were identified in 40 patients (22.3%), 79.4% (27/34) mutations occurred only once and a possible hot spot, A26 in MYH7, was found. Distribution of mutations was 52.9% (18/34) (MYBPC3), 35.3% (12/34) ( MYH7) and 11.8% (4/34) (TNNT2) respectively. Double mutations were identified in 2.2% (4/179) patients. Genotype-positive patients were associated with an earlier symptom onset, severer left ventricular hypertrophy, a higher incidence of syncope, and were more likely to have positive family history of HCM or sudden cardiac death (SCD) , and were more likely to progress into heart failure (24.2% vs. 5.0%, P = 0.002) and at a higher risk of SCD (9.1% vs. 0, P = 0.009) during the 6.5-year follow-up. No statistical difference in any clinical parameters and outcomes was found between patients carrying MYBPC3 and MYH7 mutations. Double mutations were associated with malignant clinical progression in this cohort. Different phenotype severity could be seen in HCM patients with same genotype (e. g. MYH7-1736T, TNNT2-R92W).

CONCLUSIONMYBPC3 is the most predominant gene mutation in this HCM cohort. The presence of a sarcomere mutation in patients with HCM is associated with poor clinical outcome, although no specific genes or mutations can exactly predict the severity of clinical phenotypes.

Asian Continental Ancestry Group ; Cardiomyopathy, Hypertrophic ; Carrier Proteins ; Death, Sudden, Cardiac ; Disease Progression ; Genotype ; Humans ; Hypertrophy, Left Ventricular ; Mutation ; Phenotype ; Sarcomeres ; Troponin T ; Ventricular Myosins

When the size of a neurosphere cultured in vitro reaches a certain critical value, a necrotic core will appear inside the neurosphere because of the limitation of oxygen or other nutrients transport from medium to the cells in the neurasphere. Large necrotic core will greatly reduce the expansion of NSCs. The cellular automaton (CA) model is applied in this article to model the growth of NSCs in sphere state. The appearance and enlargement of the necrotic core in a neurosphere is calculated by coupling the CA model with the nutrient diffusion analysis in bioreactors. The calculation results indicate that the culture conditions, such as seeding density, the concentration of nutrients in medium and the mass transfer coefficient between a neurosphere and medium, have some effects on the appearance of the necrotic core. However, the necrotic core mainly depends on the inner diffusion. It will certainly appear if the size of the neurosphere is large enough even the outside mass transfer is in a good condition in bioreactors. Additionally, the appearance of the necrotic core resulting from the shortage of oxygen is earlier than that caused by the limitation of glucose. And the growth of the necrotic core is very fast after its appearance, and the whole neurosphere may become necrotic. The model developed with cellular automaton and mass transfer is a good qualitative representation of NSCs growth in bioreactors.
Bioreactors ; Cell Culture Techniques ; methods ; Cell Differentiation ; Cell Proliferation ; Cells, Cultured ; Computer Simulation ; Models, Biological ; Neurons ; cytology ; Spheroids, Cellular ; Stem Cells ; cytology ; Tissue Engineering ; methods

Objective:To study the effect of miR-539-5p on apalutamide (ARN-509) sensitivity and malignant phenotype of androgen independent prostate cancer cell line C4-2B and related mechanisms.Methods:Castrated resistant prostate cancer, castrated sensitive prostate cancer and benign prostate tissue were obtained. C4-2B cell lines were divided into blank group, transfection group (miR-539-5p plasmid) and control group (control plasmid). qPCR was used to detect the expression of miR-539-5p, androgen receptor (AR) and HSBP1 in the tissues and 3 group of cells. The protein expressions of AR and HSBP1 were detected by western blot. Transwell assay was used to detect the invasion and migration ability of three groups of cells. CCK-8 assay was used to detect the proliferation ability and semi-inhibitory concentration (IC50) of AR antagonist ARN-509. The colony forming ability of the three groups of cells was detected by plate cloning experiment.Results:Tissue-qPCR indicated that, in the benign prostate tissue, tumor tissue of castration sensitive patients and tumor tissue of castration resistant patients, the expressions of miR-539-5p were 0.29 ± 0.04, 0.17 ± 0.02 and 0.07 ± 0.01, the expressions of AR were 0.13 ± 0.02, 0.28 ± 0.04 and 0.79 ± 0.11, and the expressions of HSBP1 were 0.20 ± 0.03, 0.38 ± 0.04 and 0.72 ± 0.11, respectively. Compared with benign prostate tissue and prostate cancer tissue, the expression of AR and HSBP1 gene was higher in prostate cancer tissues with castration resistance, and the expression of miR-539-5p was lower. Cell-qPCR demonstrated that the expressions of miR-539-5p in blank group, control group and transfection group were 1.00±0.09, 1.07±0.11 and 7.19±0.51, the expressions of AR were 1.00±0.10, 1.03±0.14 and 0.51±0.08, and the expressions of HSBP1 were 1.00±0.10, 0.96±0.12 and 0.97±0.11. The expression of miR-539-5p in the transfection cells was significantly higher than that in the control group and the blank group, the expression of AR gene was significantly lower than that in the control group and the blank group, and there was no significant difference in the expression of HSBP1. Western blot showed that, in blank group, control group and transfection group, the protein expressions of AR were 1.00±0.10, 1.12±0.22 and 0.72±0.16, and the expressions of HSBP1 were 1.00±0.10, 0.94±0.18 and 0.48±0.11. The protein expression of AR and HSBP1 in the transfection group was significantly lower than that in the control group and the blank group. Transwell experiment showed that the invasion and migration of cells in the transfection group were significantly lower than that in the control group and the blank group. CCK-8 assay and plate cloning experiment showed that the proliferative capacity and the number of clone formation in the transfection group were significantly lower than those in the control group and the blank group, and the expression of AR and HSBP1 in the transfection group was significantly lower than that in the control group and blank group. Compared with the control group and blank group, the IC50 value of ARN-509 decreased significantly in the transfection group.Conclusion:miR-539-5p may inhibit the malignant phenotype and castration resistance of cells via interfering with the translation level of HSBP1.

Objective:To explore and analyze the correlation between miR-31 in peripheral blood and oxidative stress indicators of diabetic nephropathy.Methods:A total of 94 patients with diabetic nephropathy who were admitted to Affiliated Hospital of Jining Medical College from September 2019 to September 2020 were selected. Patients were divided into mild diabetic nephropathy [estimated glomerular filtration rate (eGFR) 60-90 ml/min, 36 cases] group, moderate diabetic nephropathy (eGFR 30-60 ml/min, 27 cases) group and severe diabetic nephropathy (eGFR 0-30 ml/min, 31 cases) group according to the severity of the disease, and 30 healthy people in the same period were selected as the control group. Real time quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression of miR-31 in peripheral blood. Serum superoxide dismutase (SOD), malondialdehyde (MDA), advanced oxidation protein products (AOPP) and other oxidative stress indicators, as well as serum urea nitrogen, creatinine and glomerular filtration rate. Pearson was used to analyze the correlation between peripheral blood miR-31 and oxidative stress indexes and renal function.Results:The expression of miR-31 in peripheral blood of patients with diabetic nephropathy was significantly lower than that in the control group, and the expression of miR-31 in peripheral blood of patients in severe and moderate diabetic nephropathy group was significantly lower than that in the mild diabetic nephropathy group (all P<0.05), with statistically significant difference (all P<0.05). Pearson correlation analysis showed that serum miR-31 expression was negatively correlated with the severity of diabetic nephropathy ( r=-0.526, P<0.05). The levels of serum MDA, SOD and AOPP in the diabetic nephropathy group were significantly higher than those in the control group, and the levels of serum MDA, SOD and AOPP in the severe and moderate diabetic nephropathy groups were higher than those in the mild diabetic nephropathy group, with statistically significant difference (all P<0.05). The levels of serum creatinine and blood urea nitrogen in the diabetic nephropathy group were higher than those in the control group, and the glomerular filtration rate was lower than that in the control group (all P<0.05). The levels of serum creatinine and blood urea nitrogen in the severe and moderate diabetic nephropathy group were higher than those in the mild diabetic nephropathy group, while the level of glomerular filtration rate was lower than that in the mild diabetic nephropathy group, with statistically significant difference (all P<0.05). Pearson correlation analysis showed that the expression of miR-31 in peripheral blood was negatively correlated with the levels of MDA, SOD, AOPP, serum creatinine and urea nitrogen (all P<0.05), but positively correlated with glomerular filtration rate ( P<0.05). Conclusions:The expression of miR-31 in peripheral blood gradually decreases with the severity of renal damage. Its level is negatively correlated with oxidative stress indicators of diabetic nephropathy, and positively correlated with glomerular filtration rate, which can be used for for clinical treatment and disease evaluation.