Acute pancreatitis is one of the most common acute abdominal disease requiring acute hospitalization worldwide.As severe acute pancreatitis associated with mortality up to 30％,rational diagnosis and management requires up-to-date evidence-based treatment guidelines.The Guidelines for the surgical management of acute pancreatitis composed by the International Association of Pancreatology (IAP) was published in 2002,since then there have been substantial improvements in the management of acute pancreatitis.The collaboration of the IAP and the American Pancreatic Association (APA) was undertaken to revise these guidelines using an evidence-based approach in 2013 and published their new guidelines 2013 IAP/APA evidence-based guidelines for the management of acute pancreatitis few months ago.This paper presented the understandings of this new guidelines and compared with related guidelines abroad.

As the development of orthopaedics and tranmstology, treatment of bone defect is a major puzzle, and is so apparent that we need a more effective therapeusis urgently. The periosteum contains a lot of osteogenitor cells, which can self-duplicate and multi-directional differentiation. When the tissue is damaged, these cells will participate in tissue repair. In vitro cultured periosteal cells and periosteal cells injected into the small coloboma of bone have been succeeded. On the adequate biodegradable scaffolds, being regulated by some cell factors, periosteal cells can generate bone tissue that are at equal pace in tissue morphology and chemical composition of natural bone. The new bone can be juncture with vivo-bone and continue growing. It is of importance to seek a suitable scaffold materials and in vitro induction condition and to make bone formation in vitro from periosteal cells in bone tissue engineering research for clinical application.

Objective To investigate the effects ofviaminate on the proliferation and differentiation of HaCaT cells,a human keratinocyte cell line.Methods Cultured HaCaT cells were treated with various concentrations (2,5,10,15,20,25 and 30?g/mL) of viaminate for various durations.The cell proliferation was assessed by MTT method,the changes of cell cycle and apoptosis rate by flow cytometry,the changes of keratin 10 and involucrin mRNA expressions by semi-quantitative reverse transcription PCR.Results The proliferation of HaCaT cells was inhibited by the treatment with viaminate of≥2?g/mL for 48 h,and the inhibition rate was raised with the increase of treatment time and dosage.The viaminate of 30?g/mL inhibited the proliferation of HaCaT cells by 57.67% and 82.00% at 48 and 72 h after the incubation respectively,and elevated the mRNA expression of involucrin from 40.80% to 156.12%,decreased the mRNA expression of keratin 10 from 96.46% to 14.60%.The mRNA expression of involucrin increased with the elevation of viarninate dosage.Under the treatment with viaminate for 48 h,the cell population at G_1 phase significantly increased,that at S and G_2 phases decreased;the switching of G_1 to G_2 was inhibited;but the cell apoptosis was not affected.Conclusion Viaminate could inhibit the proliferation and induce the differentiation of keratinocytes.

Objective To obtain abundant human betacellulin(BTC) with biological activity. Methods The whole mature protein coding sequence of BTC gene was amplified by polymerase chain reaction(PCR) method applied to human pancreatic ?-cell tumors cDNA.The fragment was cloned into prokaryotic expression vector pET32a(+) plasmid.The recombinant plasmid was transformed into E.coli BL21 and the fusion protein was expressed under isopropyl-beta-D-thiogalactopyranoside(IPTG).The fusion protein was purified by Ni2+ affinity chromatography.SDS-PAGE and Western blot were employed to determine the expression and purification of the expected protein.BTC was added to culture NIH3T3 cells for 5 days,and cell proliferation was detected by MTT. Results Lots of fusion protein were produced,and the purified protein can stimulate the proliferation of NIH3T3 cells. Conclusion The human BTC can be successfully obtained from the pET32a(+) system with the biological activity of stimulating the proliferation of NIH3T3 cells.

Objective To construct eukaryotic expression vector of human pancreatic duodenal homeobox 1(PDX-1) gene,and to detect its expression in NIH3T3 cell lines. Methods The whole coding sequence of PDX-1 gene was amplified by polymerase chain reaction(PCR) from human pancreatic-cell tumors cDNA.The fragment was inserted into eukaryotic expression vector pcDNA3.1 plasmid.The recombinant plasmid was verified by double digestion and DNA sequencing.The expression of PDX-1 gene in NIH3T3 cells was assayed by Western blot. Results The length of specific fragment amplified by PCR was 852 bp,and the recombinant plasmid pcDNA3.1-PDX-1 showed two bands of 5.5 kb and 852 bp by digestion using respective restriction enzymes BamHⅠand EcoRⅠ.The sequence of PDX-1 gene was approved or confirmed by blasting to GenBank.It was suggested that PDX-1 gene had been cloned into pcDNA3.1 vector correctly.Western blot showed that PDX-1 gene was expressed,which was detected 24 h after pcDNA3.1-PDX-1 plasmid was transfected into NIH3T3 cells. Conclusion The recombinant eukaryotic expression vector pcDNA3.1-PDX-1 was successfully constructed and expressed in NIH3T3 cell lines.

OBJECTIVETo observe the protection of Huanglian Jiedu Decoction (HJD) on high fat diet induced liver damage mice [hyperlipidemic mice lacking apolipoprotein E (ApoE(-/-))].

METHODSWild type mice were divided into the wild common food group and the wild hyperlipidemia group. ApoE(-/-) mice were divided into the ApoE(-/-) common food group, the ApoE(-/-) hyperlipidemia group, and the ApoE(-/-) hyperlipidemia plus HJD group, 5 in each group. In the present study, wild type mice and homozygous apoE(-/-) mice were fed with a chow diet or a high cholesterol Western diet for 4 weeks. HJD at the daily dose of 5 g/kg was given to mice in the ApoE(-/-) hyperlipidemia plus HJD group by gastrogavage. The plasma levels of total cholesterol (TC), triglyceride (TG), low density cholesterol protein (LDL-C) were detected. The pathohistological changes of the liver were observed by Eosin and Hematoxylin (HE) staining. The liver macrophages and their subtype ratios, as well as macrophage surface receptor CD206 and CD36 were detected by flow cytometry.

RESULTSTypical pathological changes of simple fatty liver were manifested in the ApoE(-/-) hyperlipidemia group, TC, TG, and LDL-C increased, the macrophage ratio increased, the expression level of macrophage surface receptor CD206 decreased, showing statistical difference when compared with the ApoE(-/-) common food group (P < 0.01, P < 0.05). The ratio of alternatively activated macrophages (M2) subpopulations was lower in the ApoE(-/-) hyperlipidemia group than in the wild common food group (P < 0.05). There was no obvious change in the expression level of CD36. After intervened by HJD for 4 weeks, there was no obvious improvement in blood lipids. But the ratio of CD206+ M2 macrophages was significantly improved, when compared with the ApoE(-/-) hyperlipidemia group (P < 0.05). The pathological changes of fatty liver were significantly attenuated.

CONCLUSIONSThe liver protection effect of HJD might be associated with immunoregulation of M2 macrophage subpopulations and injured tissue repairmen. Its immunoregulation and liver protection were independent from lipids lowering.

Animals ; Apolipoproteins E ; genetics ; Cholesterol, LDL ; blood ; Diet, High-Fat ; adverse effects ; Drugs, Chinese Herbal ; pharmacology ; Fatty Liver ; metabolism ; pathology ; prevention & control ; Hyperlipidemias ; metabolism ; pathology ; prevention & control ; Liver ; cytology ; metabolism ; pathology ; Macrophages ; metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; Triglycerides ; blood

Objective Determining the characteristics of human fetal hepatocytes in vitro. Methods Isolating and culturing human fetal hepatocytes by stepwise trypsinization of liver fragment in vitro; collecting the culture medium to determine the secretion of AFP, ALB and the functional enzymes (including ALT, AST, GGT, ALP and LDH) of different generations in culture; determining the expression of cytochrome C by immunohistochemistry; testing the effects of sodium byturate on human fetal hepatocytes. Results Human fetal hepatocytes were polygonal epithelial cells in DMEM medium. They could be maintained for 5.5 months (about 30 passages) in vitro. They secreted ALB and functional enzymes all over their cultivation. Conclusion Human fetal hepatocytes can be maintained keeping function in vitro for several months.

Objective To study the effects of Zhishang Sanfang (ZS) in promoting the union of radius fracture. Methods Twenty-eight rabbit models with fr acture were randomly allocated to ZS group (Group A) and control group (Group B).His tological, histochemical and histomorphological quantitative detection was used to observe the morphological features of the frature end in the 14th and 28th da y after operation. Results The formation of blood capillaries and bone trab eculae in Group A was superior to those in Group B. The scores of the four callu s and th e depth of the lateral callus in Group A were higher than those in Group B. Con clusion ZS has obvious effect in promoting the union of fracture.

Objective To study the role of bone morphogenetic protein-7 in the osteogenic differentiation of periosteal cellsin vitro. Methods Periosteal cells, obtained from adult tibial periosteum, were cultured by routine method in vitro, and divided into two groups. One group cultured with BMP7 and the supplements of 100 nmol dexametasone, 10 mmol b-glycerophosphate and 50 mg/mL L-ascorbic acid (BMP7 group), the other cultured with the supplements alone as the control (control group). Ultrastructure and morphological changes of periosteal cells were observed by contrast phase microscope and electron microscope. In order to test the expression of markers of osteoblastic differantiation in periosteal cells, involved mineralized node and alkaline phosphatase. Each group was tested at the time of 5 d, 10 d, 15 d, 20 d, respectively, using ALP kit stain and Von Kossa stain with 3 samples at each time. Results The periosteal cells cultured by routine method and induced into osteoblast differentiation with BMP7 were both growing well, in vitro. Microscope observations showed that the periosteal cells were spindle-shaped, well-stacked, transparent and three-dimensional in the early stage, and cube-shaped or puncheon shaped in the mitotic phase, gradually became wide shuttle and irregular shape with a lot secretion in telophase. The positive cells were visible by the ALP kit staining and Von Kossa staining of calcium nodules at 5 d, 10 d, 15 d and 20 d in both groups.A difference of positive rate at each time point was found between BMP7 group and control group at 5 d, 10 d, 15 d, 20 d, and the difference was statistically significant (P ＜ 0.01). Conclusion It displayed well regeneration and osteogenesis ability in the periosteal cell. BMP7 has definite osteo-inductive activity, which can obviously enhance the proliferation and ossifyng differentiation of periosteal cells.

Background and purpose:Lung cancer is the leading cause of cancer-related deaths worldwide. Approximately 15%of all histological types consist of small cell lung cancer (SCLC). Chemotherapy is one of the major treatment methods. Though the current front-line standard chemotherapy regimen for SCLC is active in most SCLC cases, however, the disease recurs shortly after the ifrst successful treatment with multi-drug resistance phenotype. Our previously study showed that zinc ifnger protein X-linked (ZFX) was overexpressed in SCLC cells. This study aimed to investigate the expression of ZFX in SCLC tissues, and to clear their possible associations with clinical parameters and provide basis for therapy of SCLC. Methods:A total of 98 surgical specimens of small cell lung cancer were collected. The expression of ZFX was examined by quantiifcational real-time polymerase chain reaction in 78 specimens taken from patients with complete clinical data. Results:The expression of ZFX was signiifcantly increased in extensive stages than in limited stages. The expression of ZFX was associated with tumor stage, the sensitivity of chemotherapy, and survival times (all P<0.05), no data was found correlated with gender and ages (P>0.05). Conclusion: ZFX expression might be associated with the development of SCLC, and may be a potential prognosis predictor.