Objective • To establish the transforming growth factor-β2 (TGF-β2) induced epithelial-mesenchymal transition (EMT) model of retinal pigment epithelium cells, and investigate the effect and mechanism of lutein on EMT. Methods • ARPE-19 cells were cultured and divided into 4 groups including control group, TGF-β2 group, TGF-β2+lutein group and lutein group. The mRNA levels of α-smooth muscle actin (α-SMA), fibronectin (FN) and E-cadherin were analyzed by real-time PCR. The protein expression of α-SMA, FN and occludin were assayed by Western blotting. Immunofluorescence was used to detect the change of α-SMA. Meanwhile, Western blotting was performed to detect the expression levels of pSmad3 in the TGF/Smad signaling pathway. Results • TGF-β2 induced EMT was inhibited by lutein. Lutein decreased the mRNA and protein levels of the mesenchymal markers α-SMA and FN, and increased the expression of the epithelial markers E-cadherin and occludin (all P<0.05). Immunofluorescence showed that lutein can inhibit the conversion of epithelial cells into myofibroblasts. Lutein significantly downregulated the high expression of pSmad3 in TGF-β2 treated ARPE-19 cells (P=0.001). Conclusion • Lutein inhibits TGF-β2 induced EMT by downregulating the expression of pSmad3 in TGF-β/Smad signaling pathway, indicating it may attenuate subretinal fibrosis.

Studies have found that metformin is not only the preferred drug for lowering blood sugar, but also shows lipid-lowering and weight-loss effects. The purpose of this study was to use a hyperlipidemia hamster model to investigate the lipid-lowering effect of metformin and its effect on important metabolic pathways in lipid metabolism disorders. Fifty golden hamsters were divided into a control group, a model group, metformin high- and low-dose groups, and a simvastatin group. A high-fat diet was fed for 1 week to create the model, and then drug was administered for 11 weeks with the high-fat diet. Serum was taken for measurement of blood lipid and blood glucose at 2, 6, and 9 weeks after administration, and at weeks 3, 5, and 9 feces and urine were collected for ¹H NMR metabolomics tests. After 11 weeks of intravenous injection of [U-¹³C₆] glucose, serum was collected for a ¹³C NMR metabolic flux test. The results showed that the administration of metformin can significantly reduce blood lipids and glucose levels and can significantly affect metabolic pathways such as sugar metabolism, lipid metabolism, ketone metabolism, amino acid metabolism, and intestinal flora metabolism. The results of the metabolic flux analysis showed that the high-fat diet reduced the metabolism of tricarboxylic acids by 37.48%. After administration of low and high doses of metformin the metabolism of tricarboxylic acid increased by 98.14% and 143.10%, respectively. After administration of simvastatin tricarboxylic acid metabolism increased by 33.18%. The results indicate that metformin has a significant effect on promoting energy metabolism. This study used a combination of metabolomics and metabolic flow to explore the effect of metformin on lipid metabolism disorders and quantifies changes in the key pathway of energy metabolism-the tricarboxylic acid cycle. This study provides useful information for the study of the efficacy and mechanism of metformin, as well as a practical technical method for the screening of lipid-lowering drugs based on a hamster model.

OBJECTIVE: To investigate the application of ITO method and discriminant functions method in full sibling and half sibling identification. METHODS: Five hundred pairs of full siblings (FS), 50 pairs of half siblings (HS) and 500 pairs of unrelated individuals (UR) were genotyped by PowerPlex 16 system. Full sibling index (FSI), half sibling index (HSI) and the FSI:HSI ratio were calculated with ITO method. Allelic matching of each pair of the three groups was compared. The locus numbers of no-allele sharing (x0), half-allele sharing (x1) and two-alleles sharing (x2) were calculated, respectively. The discriminant functions about full-siblings, half-siblings and unrelated individuals (UR) were established by SPSS 13.0 statistical software. RESULTS: (1) Regard FSI > or = 19 or FSI < 1 as the standard of distinguishing full sibling from unrelated individual, the alternate correct percentage was 96.4%. Regard HSI > or = 19 or HSI < 1 as the standard of distinguishing half sibling from unrelated individual, the alternate correct percentage was 85.3%. Regard FSI:HSI > or = 1 or FSI:HSI < 1 as the standard of distinguishing full sibling from half sibling, the alternate correct percentage was 87.5%. (2) Four groups of discriminant functions were established. The alternate correct percentage of these discriminant functions were 84.4%-97.7%, with the highest one in full sibship-unrelated individual group. CONCLUSION Both ITO method and discriminant functions method are efficient in identification of full sibling or half sibling.
Alleles ; Discriminant Analysis ; Forensic Genetics ; Genetic Variation ; Genomic Imprinting/genetics* ; Genotype ; Humans ; Paternity ; Siblings ; Tandem Repeat Sequences/genetics*

OBJECTIVETo evaluate the safety and utility of the methods of spleen sub-pedicle two steps severance with LigaSure Vessel Sealing System combined with ultrasound scalpel to resect peri-splenic ligaments during laparoscopic splenectomy.

METHODSThe methods and the curative effect of 32 patients with laparoscopic splenectomy performed by the way as mentioned above were summarized and analyzed. 4 males, 28 females, median 36 years (range 16 - 64 years). Mean dimensions of spleens were 17 cm (range 11 - 23 cm). Nineteen patients had idiopathic thrombocytopenia purpura (ITP), 6 hereditary spherocytosis (HS) (5 of them were the same family constellation), 3 hemolytic anemia (HA), 2 spleen injury with haematoma infection, 1 Evan syndrome, 1 leukemia. Peri-splenic ligaments were resected by LigaSure combined with ultrasound scalpel, splenic pedicle was resected by spleen sub-pedicle two steps severance with LigaSure when splenic pedicle were sufficiently free. Splenic pedicle was non-excisional occluded by LigaSure at pancreatic cauda, in the first step and was occluded and resected alongside of the spleen in the second step.

RESULTSAll of the 32 cases were completely succeed, the average operating time was 70 min (55 - 130 min), the estimated intraoperative blood loss was 200 ml (50 - 600 ml), no complications such as abdominal haemorrhage, visceral injury, pancreatic leakage and infection were observed. All the patients were healed well, the average hospital stay after an operation was 6 days. The average expense of each patients could save 8050 yuan because Endo-GIA was not used.

CONCLUSIONSSpleen sub-pedicle two steps severance with LigaSure Vessel Sealing System combined with ultrasound scalpel to resect peri-splenic ligaments during laparoscopic splenectomy, which shorten the operating persistence time with less hemorrhage is a safe, effective and low-cost minimally invasive surgery in selective cases.

Adolescent ; Adult ; Blood Loss, Surgical ; prevention & control ; Female ; Humans ; Laparoscopy ; methods ; Ligaments ; surgery ; Ligation ; methods ; Male ; Middle Aged ; Retrospective Studies ; Splenectomy ; methods ; Splenic Artery ; surgery ; Splenic Vein ; surgery ; Treatment Outcome

BACKGROUNDThe prevalence of chronic pancreatitis has increased during recent years in Asia-Pacific areas as well as in China. The etiologies vary in different regions and periods. This study aimed to investigate the changing etiologies of chronic pancreatitis within 20 years at Peking Union Medical College Hospital in China.

METHODSRetrospective analysis of the etiologies of 636 cases of chronic pancreatitis at Peking Union Medical College Hospital from 1990 to 2010 was performed. Patients were divided into two groups according to two time periods (1990 - 2000 and 2001 - 2010). Statistical analysis was performed using the chi-square test.

RESULTSThe morbidity rate of chronic pancreatitis in China has recently increased. The main etiology changed from biliary diseases in the 1990s (decreased from 36.8% to 28.1%) to alcohol abuse after the year 2000 (increased from 26.5% to 36.8%). The main etiology of biliary diseases is stones in the cholecyst or bile duct, and the percentage of cholecystitis cases has increased. Autoimmune disease, including autoimmune pancreatitis, has increased quickly and currently accounts for 7.3% of cases because a greater number of autoimmune pancreatitis cases are being diagnosed. Approximately 9.5% of chronic pancreatitis cases are caused by multiple factors such as alcohol abuse and bile duct stones. Other factors include cholecystectomy and acute pancreatitis.

CONCLUSIONSThe main etiology of chronic pancreatitis has changed from biliary disease to alcohol abuse in recent years. Autoimmune factors have also obviously increased.

Adult ; Alcoholism ; complications ; Bile Duct Diseases ; complications ; China ; epidemiology ; Female ; Humans ; Male ; Middle Aged ; Pancreatitis, Chronic ; epidemiology ; etiology ; immunology ; Retrospective Studies ; Risk Factors

OBJECTIVETo evaluate the inhibitory effect of Nandeshi, an acrosin inhibitor, on human acrosin activity.

METHODSWe collected sperm samples from 10 healthy fertile men and cultured them with Nandeshi at 30 degrees C for 5 minutes at the concentrations of 0. 100, 0.120, 0.144, 0.173, 0.207, 0.249, 0.299, 0.358 and 0.430 mmol/L, with the controls treated with a well-known acrosin inhibitor N-alpha-p-tosyl-L-lysine chloromethylketone (TLCK) at 150.0, 189.8, 213.6, 240.3, 270.3, 304.1 and 342.1 mmol/L. Then we determined the residual activity of human acrosin by improved Kennedy assay.

RESULTSThe residual activity of acrosin was negatively correlated with the Nandeshi concentration, and Nandeshi exhibited an inhibition rate about 800 times that of TLCK.

CONCLUSIONNandeshi has a powerful inhibitory effect on human acrosin, and improved Kennedy assay is a simple, practical and highly sensitive technique for the detection of human acrosin activity.

Acrosin ; antagonists & inhibitors ; metabolism ; Contraceptive Agents, Female ; pharmacology ; Enzyme Inhibitors ; pharmacology ; Humans ; Male ; Spermatozoa ; drug effects ; Tosyllysine Chloromethyl Ketone ; pharmacology

Da Chaihu decoction is a classic prescription for the treatment of cholecystitis that is widely used in clinical practice, and has a definite curative effect. However, due to its diverse components and complex functions, the traditional indexes fail to capture its overall efficacy. Therefore, this study analyzed and predicted the quality markers (Q-markers) of Da Chaihu decoction based on specific chromatogram and network pharmacology to provide a reference for the comprehensive control of the quality. The study obtained 35 potential practical components of Da Chaihu decoction through virtual screening. The specific chromatogram of 15 batches of Da Chaihu decoction was established by HPLC-DAD with neohesperidin as a reference. Compared with the chromatographic peaks and the reference substance, the chemical components were assigned to predict the nine components of albiflorin, paeoniflorin, naringin, hesperidin, neohesperidin, baicalin, wogonoside, saikosaponin b₂, saikosaponin b₁ as Q-markers of Da Chaihu decoction. Finally, the network of the "components-key targets-signal pathways-biological processes" was constructed by network pharmacology to explore the mechanism of Da Chaihu decoction in treating cholecystitis to clarify the accuracy of Q-markers. The results indicated that potential Q-markers could act on multiple targets to regulate inflammatory and metabolism, and then combine to treat cholecystitis. Q-markers could combine with the pharmacologic action of Da Chaihu decoction, which could elucidate the overall efficacy of Da Chaihu decoction. This study explored the Q-markers of Da Chaihu decoction combined with the specific chromatogram and network pharmacology, which provided a basis for the quality control and evaluation of Da Chaihu decoction.

In this study, the effect of benzo[α]pyrene (BaP) on chaperone-mediated autophagy (CMA) in a simulated hypoxia environment was observed and the relationship to heat shock protein 90 (HSP90) was clarified. With HSP90 inhibitor geldanamycin (GA) and HSP90α silenced, the mRNA and protein expression of hypoxia-inducible factor-1α (HIF-1α), HSP90, heat shock cognate protein 70 (HSC70), and lysosomal associated protein 2A (LAMP-2A) of A549 cells on hypoxic environment by BaP were tested. Alkaline comet experiment, immunofluorescence γ-H2AX focus experiment, quantitative real-time PCR (qPCR), and Western blot analyses were used to clarify the relationship between the DNA damage of different concentrations of BaP in A549 cells and the mRNA and protein expression of CMA-related factors. The results show that hypoxia can promote the expression of mRNA and protein of CMA-related factors in A549 cells. This study found that BaP has an inhibitory effect on CMA under the hypoxic environment. The inhibition or silencing of HSP90 will enhance the inhibitory effect of BaP on CMA. In a normoxic environment, BaP causes DNA damage and promotes CMA.