Objective To investigate the roles of toll like receptor7 (TLR7) and toll like receptor 9 (TLR9) in the pathogenesis of acute pancreatitis.Methods AR42J cells were treated by lipopolysaccharide at different dosages (0,1,10,100 mg/L),and cell model of acute pancreatitis in vitro was established.AR42J cells without lipopolysaccharide treatment were as control.Cells and culture supernatant were collected after 24 hours cultivation.TLR7,TLR9 mRNA and protein expressions were detected by RT-PCR and Western Blot,and levels of TNF-α,IL-10 in culture supernatant were measured by ELISA.Results The TLR 7 mRNA expression levels in control group,1,10,100 mg/L lipopolysaccharide group were 0.12 ± 0.09,0.28 ± 0.06,0.49 ± 0.04,0.78 ± 0.04,and the TLR9 mRNA expression levels were 0.06 ± 0.02,0.32 ± 0.03,0.56 ± 0.14,0.84 ± 0.12; the TLR7 protein expression levels were 0.04 ± 0.01,0.26 ± 0.05,0.49 ±0.04,0.77 ±0.16,and the TLR9 protein expression levels were 0.10 ±0.14,0.62 ±0.23,1.21 ± 0.26,1.75 ± 0.13 ; the TNF-α levels in culture supernatant were (8.01 ± 5.32),(25.64 ± 8.71),(49.06 ± 10.23),(75.83 ± 6.65) ng/L,and the IL-10 levels were (155.54 ± 25.47),(105.16 ± 10.49),(69.36 ± 8.19),(14.07 ± 9.06)ng/L.The expression levels of TLR7 and TLR9's mRNA,protein in cell,as well as the levels of TNF-α in culture supernatant increased with the lipopolysaccharide concentration,while the levels of IL-10 in culture supernatant decreased with the lipopolysaccharide concentration,and the difference among these groups was statistically significant (P ＜ 0.01).Conclusions The expressions of TLR7 and TLR9 in AR42J cells treated by using lipolysaccharide are obviously up-regulated,and it suggests that TLR7 and TLR9 may be vital in the pathogenesis of acute pancreatitis.

Objective: To investigate the relationship between type 2 diabetes mellitus (T2DM) and the pathogenesis and metastasis of colorectal cancer. Methods: A case-control study was performed to compare 852 colorectal cancer patients with 940 controls (patients without cancer) recruited from 2001 to 2006, with respect to their sex, cancer subsite, the course of T2DM, hepatic metastasis, smoking and drinking. Correlated risk factors were analyzed. Results: The risk of colorectal cancer was increased in patients with T2DM and the relative risk (OR) was 2.466. The OR of male patients was higher than that of female patients, but with no significant difference (2.775 vs 2.070, P=0.394). The incidence of T2DM in patients with left hemicolon cancer was higher than that in those with right hemicolon cancer and rectal cancer, but with no significant difference between them. The colorectal cancer risk in T2DM patients with a DM course of 10 ～ 20 years was the highest, and the OR was 4.696. The rate of hepatic metastasis was higher in T2DM patients with colorectal cancer than that in celorectal cancer patients without T2DM and the OR was 2.888. Conclusion: T2DM may be one of the important pathogenic risk factors for colorectal cancer. The OR is increased with the extension of DM course within 20 years. Colorectal cancer patients with T2DM may be more prone to hepatic metastasis.

In order to solve the problem of poor communication, low efficiency of consultation, and even affecting self-confidence caused by unskilled skills and insufficient cooperation with patients in the real clinic, and to solve the problem during the clinical thinking training that beginners do not know how to organize effective information and complete the process of diagnosis and differential diagnosis more efficiently. By applying the artificial intelligence (AI) virtual patient (VP) system to the process of teaching diagnostic knowledge and clinical thinking training. It provides the students with the experience of simulating the diagnosis and treatment of the clinical real scene. Let the students talk with the VP system for inquiry training and then go to the clinic to give the real patients inquiry and by simulating the process of treating the real patients, let the students take the initiative to complete the collection of medical records and clinical decision-making under the simulated scene to train the clinical thinking. This can not only solve the shortcomings of the previous simulation teaching and clinical teaching, but also stimulate students' interest in learning. According to the results of the questionnaire, students have a high acceptance of VP system simulation teaching. Through the results of homework and assessment and evaluation, the teaching results are better than before, and this teaching method should be further popularized.

Objective To observe the expression of adrenomedullin (ADM) mRNA in hepatic tissue of rats with acute necrotizing pancreatitis (ANP) complicated with hepatic injury.Methods Sixty-four SD rats were randomly divided into control group and ANP group with 32 rats in each group.In ANP group,ANP model was induced by retrograde injection of 5％ sodium taurocholate into biliopancreatic duct of rats.Rats in control group only received sham operation and pancreas manipulation.All the rats were sacrificed at 3,6,12,24 h after the operation.The serum levels of amylase,alanine aminotransferase (ALT),aspartate aminotransferase (AST) and ADM were detected.Pathological changes in pancreatic and hepatic tissue were examined.The expressions of ADM mRNA in hepatic tissue were evaluated by fluorescence quantitative PCR.Results The serum concentrations of amylase,ALT,AST were (7229 ±968),(174.2 ±28.0),(657.7 ± 139.0) U/L,which were significantly higher than those in control group [(2036 ± 292),(104.3 ± 22.1),(419.7 ± 86.3) U/L],and the difference between the two groups was statistically significant (P ＜ 0.05 or P ＜0.01).Pathological injury of pancreas and liver tissue in ANP gradually aggravated with time,and the pathological scores at 12 h were (11.60 ± 1.51),(2.60 ± 0.89),which were significantly higher than those in control group (1.20 ± 0.77,0),and the difference between the two groups was statistically significant (P＜0.01).The serum concentrations of ADM in ANP group increased at 3 h after ANP induction,and reached (38.53 ± 6.25)pg/ml at 12 h,which was significantly higher than that in control group [(28.99 ±3.92)pg/ml] ; the concentrations of ADM in liver tissue increased at 3 h after ANP induction,and reached (3.00 ± 1.49) at 6 h,which was significantly higher than that in control group (1.04 ± 0.20),and the difference between the two groups was statistically significant (P＜0.05 or P＜0.01).Conclusions The expression of ADM mRNA in rat 's hepatic tissue increases in the early stage of ANP,and the serum concentration of ADM also increases.

BACKGROUND:Many studies have showed that enough blood supply is an essential condition of bone repair and regeneration. OBJECTIVE:To construct the endothelial progenitor cells/bone marrow mesenchymal stem cells (EPCs/BMSCs) composite sheet. METHODS:After isolation and culture, EPCs and BMSCs were co-cultured directly to form EPCs/BMSCs sheet by cellsheet-inducing medium. After 10 days of induction, the sheet was investigated by gross observation, inverted microscope and hematoxylin-eosin staining. The distribution and communication of EPCs and BMSCs during the process of cellsheet induction were observed after the fluorescence labeling separately. Alkaline phosphatase assay and alizarin red staining were applied to examine the ability of osteogenic differentiation of EPCs/BMSCs sheet.
RESULTS AND CONCLUSION:EPCs/BMSCs sheet was harvested after 10-day induction. Cel-cellcontact between EPCs and BMSCs could be observed during the process of the cellsheet preparation. The harvested sheet was composed of multiple layers of cells and cel-produced extracellular matrix. Alkaline phosphatase assay and alizarin red staining both demonstrated that EPCs/BMSCs sheet had good osteogenic differentiation ability. These results suggested that EPCs/BMSCs sheet can be constructed successful y, and the sheet has strong osteogenic differentiation capability in vitro, providing the foundation for the repair of bone defects.

BACKGROUND:Human bone marrow cells-derived extracellular matrix can promote proliferation of human periodontal ligament stem cells and maintain stem cellproperties.
OBJECTIVE:To preliminarily investigate the effect of human bone marrow cells-derived extracellular matrix on the proliferation of human periodontal ligament stem cells.
METHODS:Human periodontal ligament stem cells and bone marrow cells were separately derived from human periodontal tissue and jaw bone marrow, and human bone marrow cells-derived extracellular matrix was prepared. Human periodontal ligament stem cells were cultured and purified using limited dilution cloning method, and transmission electron microscope was used for ultrastructure observation. Human periodontal ligament stem cells at passage 2 were cultured with human bone marrow cells-derived extracellular matrix and normal culture medium (control group). The cellcounting kit-8 and flow cytometry were used to determine the proliferation potential of human periodontal ligament stem cells cultured on human bone marrow cells-derived extracellular matrix.
RESULTS AND CONCLUSION:Compared with the control group, human periodontal ligament stem cells cultured on human bone marrow cells-derived extracellular matrix had a superior capacity of proliferation (P<0.05), and the cells met their morphological and biological characteristics, and grew in good conditions. Human bone marrow cells-derived extracellular matrix is a promising matrix for large-scale expanding human periodontal ligament stem cells for future use in stem cel-based therapy.

AIM: To explore a new compute tomography (CT) for the localization of intraocular foreign bodies (IOFBs).METHODS: After CT ocular horizontal and ocular axial scan,the foreign bodies on the eyeball wall or retinal surface in 26eyes were localized by means of the combination of CT ocular axial scan and meridian plane reconstruction (new method),ocular horizontal scan and ocular axial scan (conventional method Ⅰ) as well as ocular horizontal scan right angle coordination (conventional method Ⅱ) separately. According to the criteria of indirect ophthalmoscope localization and direct observation during operation, the relative accuracies of corresponding points of the foreign bodies on sclera surface along meridian and latitude were measured.RESULTS: In the 26 cases, the mean relative accuracies of corresponding points of foreign bodies on sclera surface along meridian and latitude were 1.53mm、1.64mm (new method), 1.37mm、1.64mm (conventional method Ⅰ) and 2.02mm、2.55mm (conventional method Ⅱ) respectively.There was no statistical difference between the new method and the conventional method Ⅰ, whereas there was statistical difference between the new methods and the conventional method Ⅱ (along meridian: P＜0.05; along latitude:P＜0.01).CONCLUSION: Compared with the conventional methods,the new method is simpler, more visible and more potential in the clinic.

OBJECTIVE: To investigate the effects of Xiehuo Bushen Decoction (XHBSD), a compound Chinese herbal medicine, on the survival and differentiation of transplanted neural stem cells (NSCs) in brains of rats with intracerebral hemorrhage, and to explore the mechanism of Xiehuo Bushen formula in promoting the survival of transplanted NSCs. METHODS: NSCs separated from hippocampuses of neonatal SD rats were cultured. Sixty-five panel reactive antibody (PRA) positive SD rats were selected by lymphocytotoxicity methods. The PRA positive rats were made into intracerebral hemorrhagic model and divided into three groups: cerebral hemorrhage group (n=15), NSCs transplanted group (n=25) and XHBSD group (n=25). XHBSD was orally administered after 5-bromodeoxyuridine (BrdU)-marked NSCs were transplanted in brains of rats with intracerebral hemorrhage in the XHBSD group. Rats in the other two groups were administered distilled water. The expressions of interferon gamma (IFN-gamma) and interleukin-4 (IL-4) mRNAs were measured by reverse transcription polymerase chain reaction (RT-PCR); the numbers of BrdU and 200 kD neurofilament (NF200) positive cells were detected by double-labeling immunofluorescence method. RESULTS: The expression of IFN-gamma mRNA was down-regulated significantly in the XHBSD group, but the expression of IL-4 mRNA was up-regulated significantly (P<0.05). The numbers of BrdU and NF200 positive cells were also increased remarkably in the XHBSD group. CONCLUSION: XHBSD can promote the survival and differentiation of transplanted NSCs, which may be related to inducing the expression of IL-4 mRNA and inhibiting the expression of IFN-gamma mRNA.

Aim To investigate the effect of intestinal cholesterol absorption inhibitor Ezetimibe on lipid accumulation in RAW264.7 cells and identify the underlying mechanism.Method RAW264.7 cells were pretreated with the indicated concentrations of Ezetimibe (0,0.003,0.01 and 0.03 mol·L~(-1))for 24 hours or pretreated with the optimal concentration(0.03 mol·L~(-1))of Ezetimibe for different periods (0,6,12 and 24 h),followed by incubation with 50 mg·L~(-1) oxLDL for 24 hours,then the number of intracellular lipid droplets and lipid content were measured by using oil red O staining and HPLC; the expression of NPC1L1 was measured by Western blot.Results Pretreatment with indicated concentrations of Ezetimibe caused a concentration-dependent inhibition of intracellular lipid accumulation;pretreatment with 0.03 mol·L~(-1) Ezetimibe caused a time-dependent inhibition of intracellular lipid accumulation.It was noted that pretreatment with 0.03 mol·L~(-1) Ezetimibe for 24 hours inhibited CE by about 47%+0.1% compared with control group(oxLDL alone).Immunoblotting results showed that NPC1L1 was expressed in RAW264.7 cells and it was down-regulated after Ezetimibe treatment.Conclusions Ezetimibe causes concentration-dependent and time-dependent inhibition of lipid accumulation in RAW264.7 cells;it also reduces NPC1L1 expression in RAW264.7 cells.

BACKGROUND: The shape and amount of bone regenerated depend on the existence and maintenance of hypolemmal space. Collapse and reduction of hypolemmal space will greatly inhibit bone regeneration. Therefore, the key to success of induced bone regeneration is to select membrane material.OBJECTIVE: This study is designed to investigate the feasibility of poly (hydroxybutyrate-hydroxyvalerate) (PHBV) membrane for guiding bone regeneration.DESIGN, TIME AND SETTING: This study, a self-control animal experiment, was performed at the Laboratory Animal Center, Third Hospital Affiliated to Sun Yat-sen University between May 2005 and October 2006.MATERIALS: PHBV membrane, (10×10×0.3)mm, was provided by Research of Biomaterials, College of Material Science and Engineering, South China University of Technology. Eight healthy hybrid dogs, aged 1 year old, weighing 10-12kg, were included for model of tibial defects.METHODS: Four bone defects were made on bilateral tibias in each dog. Bilateral proximal bone defects of all dogs were included as control group and only subjected to periosteal flap reduction. Bilateral distal bone defects of all dogs were included as experimental group and covered by PHBV membrane. Samples were harvested at weeks 2, 4, 8, and 12 post-surgery.MAIN OUTCOME MEASURES: In vivo degradation performance and ossific capability of PHBV copolymer as well as energy-dispersive X-ray analysis and elemental quantitative assay in bone defect regions.RESULTS: In the experimental group, at week 8 post-surgery, PHBV copolymer membrane had been gradually degraded, different sizes of hollows formed on its surface, bone defect surface was smooth and flat, and X-ray results showed increased bone density in the bone defect regions; at week 12 post-surgery, the PHBV copolymer was still not degraded completely, but bone defect regions had been completely fused with normal bone. Energy-dispersive X-ray analysis and elemental quantitative assay results demonstrated in the experimental group, peaks of Ca and P atom number of newly formed bone were predominant, and peaks of impurity were few. With time going, element Ca and P levels in the newly formed bone were increased, the ratio for Ca and P atoms approximated to that in the normal bone. There was significant difference between the. experimental group and the control group (P<0.05).CONCLUSION: PHBV copolymer membrane is a promising ideal natural membrane material for guiding bone regeneration.