Objective　To explore the significance of the dynamic changes of pro-and anti-inflammatory cytokines in the onset and development of acute panreatitis (AP). Methods　Pro-inflammatory cytokines TNFα, IL-1β, IL-6, and IL-8 and anti-inflammatory cytokines IL-10 and IL-1ra in the plasma of 48 patients with AP and 20 healthy individuals were determined with ELISA. Results　The levels of all pro-and anti-inflammatory cytokines in plasma was significantly higher in AP patients than in control group (P<0.05) in early stage of the disease, and then all levels were decreased gradually, consistent with the alterations of clinical symptoms of the AP patients. Conclusion　The dynamic changes of pro-and anti-inflammatory cytokines might play important role in the onset and development of AP.

Objective To investigate the effect of Ginkgolide B (BN52021) on lipopolysaccharide (LPS) induced pancreas microvascular endothelialv (MS1) cells, and to explore its molecular mechanism.Methods The optimal concentration and best time point of LPS inhibing MS1 cell survival and the optimal concentration of BN52021 increasing survival of LPS induced MS1 cells were determined by MTT.The mRNA and protein expression of adenylate cyclase (AC), phospholipase A2 (PLA2), phospholipase Cβ (PLCβ),protein tyrosine kinase (PTK) and G protein coupled receptor kinase (GRK) in platelet activating factor receptor(PAFR) signal pathway in MS1 cells were determined by real-time PCR and Western blot.Results It was showed that 10 μg/ml LPS for 24 h was the optimal concentration and best time point to induce the decrease of MS1 cells.50 mmol/L of BN52021 was the optimal concentration of increacing survival of LPS induced MS1 cells.After LPS induction, AC, GRK, PLA2, PLCβ, PTK mRNA expressions of MS1 cells were 4.02 ±0.14, 2.63 ± 0.03, 3.31 ± 0.12, 2.09 ± 0.08, 1.85 ± 0.07, which were significantly higher than those in control group (P ＜ 0.01).After BN52021 treatment, AC, GRK, PLA2, PLCβ mRNA expressions of LPS induced MS1 cells were 2.35 ±0.13, 1.17 ±0.14, 1.87 ±0.11, 1.65 ±0.10, which were significantly lower than those in LPS induction group (P ＜ 0.01).The expression of PTK mRNA was 1.83 ± 0.13, which was not significantly different from that in LPS induction group.Western blot showed that the levels of protein expression were consistent with those of mRNA expression.Conclusions BN52021 can down-regulate the up-regulated genes expression of AC, GRK, PLA2 and PLCβ in the PAFR signal pathway in LPS induced MS1 cells.

Signal transducer and activator of transcription 3 (STAT3), an acute-phase response protein, is ac-tivated to over-express by cytokines .STAT3 also acts as a transcriptional factor to regulate the expression of cytokines .O-ver-expression of cytokines is accompanied by STAT 3 activation and over-expression in acute pancreatitis .Meanwhile , the proliferation of pancreatic stellate cells in chronic pancreatitis is mediated by STAT 3.In this review, the research progress in STAT3 function is summarized to elaborate its potential role in the pathogenesis of pancreatitis .

Objective:To design a kind of head holder for small animal imaging without artifact. Methods: The head holder was designed for small animal imaging according to rodent head stereotaxic apparatus with lightweight and easy molding material (polymethyl methacrylate). The holder can be easily made with ordinary lathe and can be easily operated in small animal imaging device. Results:After the trial in small animal PET-CT, the holder can effectively solve the problems in small animal imaging such as the artifact caused by tiny displacement, tilt head position, deviating from the central field of view and respiratory depression. Conclusion: The small animal imaging device was designed in simple structure to overcome the shortcomings of the existing technology. The operation of the device is simple, which is worthy of popularization head holder of application.

Objective To investigate the inhibitory effect of ginkgolide B (BN52021) on severe acute pancreatitis (SAP) via detecting the antagonistic effect of BN52021 on platelet-activating factor (PAF). Methods One hundred and eighty Wistar rats were randomly divided into control group (n = 60), SAP group (n = 60) and BN52021 group (n =60) according to the random number table. The 3 groups were divided into 6 subgroups at different time points after operation (1 h, 2 h, 3 h, 6 h, 12 h, and 24 h). The changes of serum amylase in each group were monitored. The expression of platelet-activating factor receptor (PAFR) mRNA and protein was detected by RT-PCR and Western blot, and the pathological changes of pancreatic tissues were observed. All the data were analyzed by one-way ANOVA. Results Serum amylase level and pathological results showed that it was successful in preparing SAP model. The serum amylase levels at postoperative hour 3, 6 and 24 were (4185 ±148) U/L, (3785 ± 124) U/L and (1360 ± 161) U/L in BN52021 group, which were significantly lower than those in SAP group [(4799 ± 107) U/L, (4920 ± 140) U/L, (2283 ± 127) U/L)]. The pathological scores at postoperative hour 3, 6, 12 were 5.95±0. 19, 5.55±0.36, 6.72±0. 30 in BN52021 group, which were significantly lower than those in SAP group (8.85 ± 0.39, 9.15 ± 0.55, 10.10 ±0. 65). The mRNA and protein expression of PAFR were gradually increased at the early stage (0.49 ± 0.09-0.71 ± 0.14 vs 0. 43 ～ O. 06-1.69 ± 0.06), and reached peak at postoperative hour 3. The expression levels of PAFR mRNA and protein in BN52021 group and SAP group at postoperative hour 3 had statistical difference among the 3 groups (F = 4.58, 6.24, P < 0.05). Conclusions The expression of PAFR mRNA and protein in the pancreatic tissue of SAP rats is dynamically changing. PAFR plays an important role in the occurrence and progression of SAP. BN52021 can reduce the expression of serum amylase and improve the pancreatic pathological changes, but it has no effect on the expression of PAFR in pancreatic tissue.

BACKGROUND: Ischemic preconditioning (IPC) can induce endogenous protection mechanism, which effectively prevent ischemia/reperfusion injury following organ transplantation. Cold and warm ischemia may induce ischemia/reperfusion injury of pancreas transplantation, and apoptosis of pancreatic acinar cells is one of the important reasons of pancreas graft functional defect after transplantation. Mitochondrial DNA has repair system, and its balance with mitochondrial DNA injury influences disease occurrence and outcome.OBJECTIVE: To observe the effect of IPC on apoptosis of transplanted pancreatic acinar cells, and the possible role of reactive oxygen (ROS) and mitochondrial DNA repair enzyme.METHODS: A total of 50 health, male, Sprague-Dawley rats were randomly divided into three groups: sham operated (n = 10), donors (n = 20) and recipients (n = 20). The recipients were randomly divided into ischemia/reperfusion group (IR, n = 10) and IPC group (n = 10). The sham operated group was subjected to abdominal open and close operation. IR group and IPC group received establishment of diabetic model by streptozotocin injection. IR rats received whole pancreatic-duodenal transplantation alone. IPC rats received whole pancreatic-duodenal transplantation exposed ischemic preconditioning with 5 minutes ischemia and 5 minutes reperfusion twice. All grafts were keep with warm ischemia time 15 minutes and cold ischemia (in 4 ℃ UW preservation solution) time 180 minutes. Twelve hours after reperfusion, serum amylase, blood glucose, Caspase-3, -9 activity were detected. Pancreatic acinar cell apoptosis was measured by flow cytometry. Mitochondrial cross-membrane potential (Δψ) was measured by monitoring the fluorescence spectrum of rhodamine 123. Mitochondrial H2O2 generation was determined using dichlorofluorescein as a probe. 8-oxodG in mitochondrial DNA (mtDNA) was measured with HPLC system. Release of cytochrome C, phosphorylation of Akt and mitochondrial OGG1 protein expression were determined by Western-blotting. RESULTS AND CONCLUSION: The ischemia preconditioning can relieve the pancreatic acinar cell apoptosis in pancreas graft and relieve IR injury by decreasing mitochondrial oxidative stress, mtDNA injury, and increasing phosphorylation of Akt and mitochondrial OGG1 expression.

Objective To clarify whether oxymatrine ( OM) could suppress the activation of pancreatic stellate cells ( PSC) and explore the potential molecular mechanism .Methods The proliferation of PSC line LTC 14 being activated by TGF-β1 with OM treatment at different concentrations (OM group) was measured. SOD level was determined by ELISA and p 38-MAPK mRNA was determined by real-time PCR.Results The proliferation of PSC in the control group , 0.1, 0.5, 1, 2, 5 g/L OM group was (1.51 ±0.08), (1.50 ± 0.07), (1.15 ±0.04), (1.15 ±0.04), (1.08 ±0.06), and (1.08 ±0.10), respectively.The level of the control group was lower than the groups where the concentration of OM reached or exceeded 0.5mg/ml ( all P=0.000).SOD level of LTC 14 cells in the control group, TGF-β1 group, 0.5 and 1 g/L OM group was (0.087 ±0.005), (0.073 ± 0.004), (0.085 ± 0.010), and (0.086 ± 0.007), respectively. No statistically significant difference existed among the groups (P=0.095).The p38-MAPK mRNA expression of PSC in the control group, TGF-β1 group, 0.5, and 1 g/L OM group was (1.000 ±0.000), (1.979 ± 0.505), (0.606 ±0.111), and (0.303 ±0.159), respectively.The p38-MAPK mRNA level of TGF-β1 group was higher than that of the control group (P=0.002), and that of 0.5 mg/ml OM group and 1 mg/ml OM group was lower that of TGF-β1 group ( P=0.000 ) , while no statistical difference was found between 0.5 mg/ml OM group and 1 mg/ml OM group.Conclusions OM could suppress the activation of PSC in vitro and the suppression of p38-MAPK mRNA expression may be involved .

Toll-like receptor 4 (TLR4) plays an important role in inflammation and immune response.MicroRNAs (miRNAs) are involved in the regulation of TLR4 signaling pathway in multiple levels and various molecules,which play an important role in inflammatory reaction.A variety of miRNAs are involved in the regulation of TLR4 signaling pathway,and the TLR4 signaling pathway can induce a variety of miRNAs.Chronic diseases such as diabetes,Alzheimer's disease and cardiovascular disease are closely related to inflammatory response.The regulatory role of TLR4 signaling related miRNAs has attracted much attention in inflammatory diseases.In this review,the research progress of TLR4 signaling pathway related miRNAs in the regulation of inflammatory response is summarized,which provides a new research direction for the clinical diagnosis and treatment of inflammatory response related diseases.

Although empirically well understood in their clinical administration, volatile anesthetics are not yet well comprehended in their mechanism studies. A major conundrum emerging from these studies is that there is no validated model to assess the presumed candidate sites of the anesthetics. We undertook this study to test the hypothesis that the single-celled Paramecium could be anesthetized and served as a model organism in the study of anesthetics. We assessed the motion of Paramecium cells with Expert Vision system and the chemoresponse of Paramecium cells with T-maze assays in the presence of four different volatile anesthetics, including isoflurane, sevoflurane, enflurane and ether. Each of those volatiles was dissolved in buffers to give drug concentrations equal to 0.8, 1.0, and 1.2 EC50, respectively, in clinical practice. We could see that after application of volatile anesthetics, the swimming of the Paramecium cells was accelerated and then suppressed, or even stopped eventually, and the index of the chemoresponse of the Paramecium cells (denoted as I ( che )) was decreased. All of the above impacts were found in a concentration-dependent fashion. The biphasic effects of the clinical concentrations of volatile anesthetics on Paramecium simulated the situation of high species in anesthesia, and the inhibition of the chemoresponse also indicated anesthetized. In conclusion, the findings in our studies suggested that the single-celled Paramecium could be anesthetized with clinical concentrations of volatile anesthetics and therefore be utilized as a model organism to study the mechanisms of volatile anesthetics.

Objective To explore the influence of LPS treatment on related molecules in Smads and ERK1/2 signal pathway in pancreatic stellate cell line LTC-14.Methods LTC-14 cells were cultured in vitro, and were treated with LPS at different dose in different time points.Protein expressions of related molecules in Smads pathway and ERK1/2 pathway and α-SMA in LTC-14 Cells were examined by Western blot.Results On Treated LTC-14 cells by 0, 1, 5, 10, 20 and 50 mg/L LPS,protein expressions of Smad3 were 0.15±0.02, 0.37±0.02, 0.44±0.01, 0.46±0.02, 0.372±0.01 and 0.24±0.03;expressions of Smad7 were 0.79±0.05, 0.84±0.02, 0.55±0.03, 0.45±0.03, 0.34±0.02 and 0.92±0.07;p-ERK1/2 levels were 0.48±0.05, 0.74±0.03, 0.72±0.04, 0.89±0.02, 0.81±0.02 and 0.72±0.03;p-cPLA2 levels were 0.15±0.03, 0.30±0.01, 0.31±0.01, 0.30±0.02, 0.28±0.03 and 0.32±0.02;α-SMA levels were 0.56±0.06, 0.62±0.06, 0.54±0.04, 1.03±0.11, 1.39±0.08 and 1.28±0.10.The changes of protein expressions before and after LPS treatment were obvious (all P<0.01).The protein expressions of ERK1/2 were 0.56±0.03, 0.57±0.02, 0.53±0.02, 0.58±0.02, 0.59±0.05 and 0.55±0.04, which did not change obviously along with increased LPS dosages.LTC-14 cells treated with 10 mg/L LPS for 0, 1, 3, 6 and 9 h,the expressions of Smad3 were 0.69±0.05, 0.68±0.07, 1.02±0.14, 1.82±0.0 and 2.04±0.11,those of Smad7 were 2.77±0.10, 1.37±0.08, 1.45±0.14, 0.78±0.09 and 0.63±0.06,those of p-ERK1/2 were 0.16±0.03, 0.32±0.05, 0.79±0.03, 1.50±0.07 and 1.77±0.04,those of p-cPLA2 were 0.15±0.04, 0.32±0.06, 0.63±0.04, 0.95±0.04 and 1.49±0.10,those of α-SMA were 0.84±0.03, 1.26±0.21, 1.81±0.19, 4.28±0.26 and 4.37±0.15, all of which changed obviously as the treatment time increased (P<0.05 or 0.01).The expressions of ERK1/2 were 0.75±0.03, 0.72±0.02, 0.80±0.04, 0.74±0.03 and 0.85±0.09, which did not change obviously as the treatment time increased.Conclusions LPS could upregulate the expression of α-SMA in a time-and dose-dependent way, and activate intracellular Smads and ERK1/2 inflammatory pathways, which may be the potential molecular mechanism of the development of chronic pancreatitis.