Exploring the links between gut microbiome changes and irritable bowel syndrome in Han populations in the Tibetan Plateau / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

The gut microbiome shows changes under a plateau environment, while the disbalance of intestinal microbiota plays an important role in the pathogenesis of irritable bowel syndrome (IBS); however, the relationship between the two remains unexplored. In this work, we followed up a healthy cohort for up to a year before and after living in a plateau environment and performed 16S ribosomal RNA (rRNA) sequencing analysis of their fecal samples. Through evaluating the participants' clinical symptoms, combined with an IBS questionnaire, we screened the IBS sub-population in our cohort. The sequencing results showed that a high-altitude environment could lead to changes in the diversity and composition of gut flora. In addition, we found that the longer the time volunteers spent in the plateau environment, the more similar their gut microbiota composition and abundance became compared to those before entering the plateau, and IBS symptoms were significantly alleviated. Therefore, we speculated that the plateau may be a special environment that induces IBS. The taxonomic units g_Alistipes, g_Oscillospira, and s_Ruminococcus_torques, which had been proved to play important roles in IBS pathogenesis, were also abundant in the IBS cohort at high altitudes. Overall, the disbalance of gut microbiota induced by the plateau environment contributed to the high frequency of IBS and the psychosocial abnormalities associated with IBS. Our results prompt further research to elucidate the relevant mechanism.

Effects of oxymatrine on the expression and release of high mobility group box 1 in pancreatic acinar cells stimulated by hydrogen peroxide / 中华胰腺病杂志

Objective@#To observe the effects of oxymatrine(OM) on the expression and release of high mobility group box 1(HMGB1) in rat pancreatic acinar cell line AR42J stimulated by hydrogen peroxide.@*Methods@#MTT method was used to detect the effects of H2O2 in different concentrations on the survival of AR42J cells. AR42J cells cultured in vitro were divided into control group, H2O2 group and H2O2+ OM group. An equal volume of H2O2(final concentration 0.16 mmol/L) was added in H2O2 group and H2O2+ OM group, respectively, while an equal volume of triple distilled water was added in control group. In H2O2+ OM group, OM(final concentration 0.5 g/L)was added 0.5 h before the addition of H2O2, and cell samples and supernatant were collected after 24 h culture. The expression of HMGB1 protein was detected by Western blotting, the level of HMGB1 protein in cell supernatant was detected by enzyme-linked immunosorbent assay, and the intracellular distribution of HMGB1 protein was detected by immunofluorescence.@*Results@#In the H2O2 group, the expression of HMGB1, the secretion of HMGB1 in the supernatant and the proportion of cytoplasmic HMGB1 in the total HMGB1 were significantly higher than those in the control group[1.04±0.04 vs 0.69±0.02, (4.84±0.13)μg/L vs (2.68±0.07)μg/L, (35.7±2.5)% vs (10.7±1.9)%], and all the differences were statistically significant (all P<0.05). After OM intervention, HMGB1 protein expression, secretion and cytoplasmic proportion were [0.82±0.02, (3.97±0.10)μg/L and (27.3±1.7)%], respectively, which were obviously lower than those in the H2O2 group, and all the differences were statistically significant (all P<0.05).@*Conclusions@#H2O2 can induce the expression and release of HMGB1 in rat pancreatic acinar cells; OM treatment could alleviate the severity of oxidative stress injuries induced by H2O2 in AR42J cells.

Effects of oxymatrine on the expression and release of high mobility group box 1 in pancreatic acinar cells stimulated by hydrogen peroxide / 中华胰腺病杂志

Objective:To observe the effects of oxymatrine(OM) on the expression and release of high mobility group box 1(HMGB1) in rat pancreatic acinar cell line AR42J stimulated by hydrogen peroxide.Methods:MTT method was used to detect the effects of H 2O 2 in different concentrations on the survival of AR42J cells. AR42J cells cultured in vitro were divided into control group, H 2O 2 group and H 2O 2+ OM group. An equal volume of H 2O 2(final concentration 0.16 mmol/L) was added in H 2O 2 group and H 2O 2+ OM group, respectively, while an equal volume of triple distilled water was added in control group. In H 2O 2+ OM group, OM(final concentration 0.5 g/L)was added 0.5 h before the addition of H 2O 2, and cell samples and supernatant were collected after 24 h culture. The expression of HMGB1 protein was detected by Western blotting, the level of HMGB1 protein in cell supernatant was detected by enzyme-linked immunosorbent assay, and the intracellular distribution of HMGB1 protein was detected by immunofluorescence. Results:In the H 2O 2 group, the expression of HMGB1, the secretion of HMGB1 in the supernatant and the proportion of cytoplasmic HMGB1 in the total HMGB1 were significantly higher than those in the control group[1.04±0.04 vs 0.69±0.02, (4.84±0.13)μg/L vs (2.68±0.07)μg/L, (35.7±2.5)% vs (10.7±1.9)%], and all the differences were statistically significant (all P<0.05). After OM intervention, HMGB1 protein expression, secretion and cytoplasmic proportion were [0.82±0.02, (3.97±0.10)μg/L and (27.3±1.7)%], respectively, which were obviously lower than those in the H 2O 2 group, and all the differences were statistically significant (all P<0.05). Conclusions:H 2O 2 can induce the expression and release of HMGB1 in rat pancreatic acinar cells; OM treatment could alleviate the severity of oxidative stress injuries induced by H 2O 2 in AR42J cells.

Research progress of TLR4 signaling pathway related miRNAs in inflammatory response / 天津医药

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.

The influence of LPS on the protein expression of related molecules in Smads and ERK1/2 signal pathway in LTC-14 cells / 中华胰腺病杂志

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.

STAT3 signaling in pathogenesis of pancreatitis / 中国病理生理杂志

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 .

Oxymatrine suppressed the activation of pancreatic stellate cells and p38-MAPK mRNA expression in vitro / 中华胰腺病杂志

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 .

Design of artifact-free head holder for small animal imaging / 中国医学装备

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.

Mechanism of effect of Ginkgolide B on lipopolysaccharide induced microvascular endothelial cells / 中华胰腺病杂志

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.

Ischemic precondition inhibits pancreatic acinar cells apoptosis in rats with ischemia/reperfusion injury following pancreas transplantation: Role of reactive oxygen and mitochondrial DNA repair enzyme / 中国组织工程研究

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.

Effects of ginkgolide B (BN52021) on pathological changes Of pancreatic tissue in rats with severe acute pancreatitis / 中华消化外科杂志

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.

Dynamic changes of pro- and anti-inflammatory cytokines in acute pancreatitis / 第三军医大学学报

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.