ObjectiveTo investigate the anti‑pancreatic fibrosis mechanism of Klotho‑derived peptide 7 （KL7） by observing its effect on a mouse model of chronic pancreatitis （CP） induced by cerulean， and to provide a basis for clinical medication. MethodsA total of 40 male BALB/c mice were randomly divided into control group， model group， low-dose KL7 group （2 mg/kg）， and high-dose KL7 group （4 mg/kg）， with 10 mice in each group. All mice except those in the control group were given intraperitoneal injection of cerulean （50 μg/kg） 6 times a day at an interval of 1 hour， twice a week for 4 consecutive weeks to establish a model of CP. The mice in the low-dose KL7 group and the high-dose KL7 group were treated with different doses of KL7 once a day for 4 consecutive weeks. In vivo imaging was used to observe the accumulation of KL7 in the pancreas； molecular docking was used to detect the binding of KL7 to transforming growth factor-β type Ⅱ receptor （TβRⅡ）； the mice were measured in terms of body weight and pancreatic weight； HE staining was used to observe the pathological changes of pancreatic tissue； Masson staining was used to observe the degree of pancreatic fibrosis； immunohistochemical staining was used to measure the expression of α-smooth muscle actin （α-SMA） and type Ⅰ collagen （COL1A1）； Western blotting was used to measure the protein expression levels of α-SMA， TβRII， and phosphorylated small mothers against decapentaplegic homolog 2/3 （p-Smad2/3） in pancreatic tissue. A one-way analysis of variance was used for comparison of continuous data between multiple groups， and the least significant difference t-test and the Dunnett’s-T3 test were used for further comparison between two groups. ResultsKL7 was significantly enriched in the pancreatic tissue of CP mice， and there was a strong binding activity between KL7 and TβRⅡ. Compared with the control group， the model group had significant reductions in pancreatic mass and relative pancreatic mass （P<0.000 1）， with disordered structure of pancreatic tissue， an increase in inflammatory cell infiltration， and significant increases in fibrosis degree， the positive areas of α-SMA and COL1A1 （P<0.000 1）， and the protein expression levels of α-SMA， TβRⅡ， and p-Smad2/3 （P<0.05）. Compared with the model group， the high-dose KL7 group had significant increases in pancreatic mass and relative pancreatic mass （P<0.01）， with alleviation of structural damage of pancreatic tissue and inflammatory cell infiltration， a significant reduction in fibrosis degree， and significant reductions in the positive areas of α-SMA and COL1A1 （P<0.001） and the protein expression levels of α-SMA， TβRⅡ， and p-Smad2/3 （P<0.01）. ConclusionKL7 has a significant targeted therapeutic effect on pancreatic fibrosis in CP mice through specific binding of KL7 to TβRⅡ， thereby inhibiting the activation of the TGF-β/Smad signaling pathway.

Objective:To discuss the inhibitory effect of schisandrin B(SchB)on the migration and invasion of the pancreatic cancer Pan02 cells,and to clarify its mechanism.Methods:The pancreatic cancer Pan02 cells were treated with different concentrations of SchB(0,0.78,1.56,3.12,6.25,12.50,and 25.00 mg·L-1)for 24,48,and 72 h.CCK-8 method was used to detect the survival rates of the cells in various groups,and the concentration of SchB for the subsequent experiments was confirmed.The Pan02 cells were divided into control group,2.5 mg·L-1 SchB group,5.0 mg·L-1 SchB group,and 10.0 mg·L-1 SchB group.Wound healing assay was used to detect the wound healing rates of the Pan02 cells in various groups;Transwell chamber assay was used to detect the numbers of migration and invasion Pan02 cells in various groups;Western blotting method was used to detect the expression levels of Vimentin and N-cadherin proteins in the Pan02 cells in various groups.The mouse models of subcutameous transplanted tumor of pancreatic cancer cells were established.Ten successfully modeling mice were randomly divided into control group and SchB group(n=5).After 28 d of treatment,the weights of tumor of the mice were determined;immunohistochemistry method was used to detect the expressions of Vimentin and N-cadherin proteins in tumor tissue of the mice in various groups.Results:The CCK-8 results showed that compared with control group,the survival rates of the pancreatic cancer Pan02 cells in different concentrations of SchB groups were decreased(P＜0.05 or P＜0.01).The wound healing results showed that compared with control group,the wound healing rates of the cells in 2.5,5.0,and 10.0 mg·L-1 SchB groups were decreased(P＜0.05 or P＜0.01).The Transwell chamber results showed that compared with control group,the numbers of migration and invasion Pan02 cells in 2.5,5.0,and 10.0 mng·L-1SchB groups were decreased(P＜0.05 or P＜0.01).The Western blotting results showed that compared with control group,the expression levels of Vimentin and N-cadherin proteins in the Pan02 cells in 2.5,5.0,and 10.0 mg·L-1 SchB groups were decreased(P＜0.05 or P＜0.01).Compared with control group,the tumor volume and weight of the mice in SchB group were significantly decreased(P＜0.01).The immunohistochemistry results showed that compared with control group,the positive expression rates of Vimentin and N-cadherin proteins in tumor tissue of the mice in SchB group were significantly decreased(P＜0.01).Conclusion:SchB can inhibit the proliferation,migration,and invasion of the pancreatic cancer Pan02 cells,and its mechanism is related to the reduction of expressions of Vimentin and N-cadherin proteins.

Objective:To discuss the inhibitory effect of Schisandrin B on the proliferation of pancreatic cancer Pan02 cells,and to clarify the mechanism.Methods:CCK-8 method was used to detect the proliferation rates of the Pan02 cells after treated with different concentrations(0,0.78,1.56,3.12,6.25,12.50,and 25.00 mg·L-1)of Schisandrin B to select the optimal concentration and treatment time of Schisandrin B.The mouse pancreatic cancer Pan02 cells were divided into control group(0 mg·L-1 Schisandrin B),2.5 mg·L-1 Schisandrin B group,5.0 mg·L-1 Schisandrin B group,and 10.0 mg·L-1 Schisandrin B group.The morpholoy of Pan02 cells invarious groups was observed with light microscope;5-ethynyl-2'-deoxyuridine(EdU)staining assay was used to detect the positive expression rates of the Pan02 cells in various groups;flow cytometry was used to detect the percentages of the Pan02 cells at different cell cycles and the apoptotic rates of the cells in various groups;Western blotting method was used to detect the expression levels of cell cycle and apoptosis-related proteins in the cells in various groups.Results:The CCK-8 method results showed that after treated with Schisandrin B for 48 and 72 h,compared with 0 mg·L-1 Schisandrin B,the proliferation rates of the Pan02 cells after treated with different concentrations of Schisandrin B were decreased(P<0.01),especially at 72 h.0.25,5.0,and 10.0 mg·L-1 Schisandrin B were selected to treat the Pan02 cells,and 72 h was the treatment time.In control group,the Pan02 cells had a spindle shape,with good condition,and grew closely adhered to the wall with normal organelles and cytoplasm,in 2.5 and 5.0 mg·L-1 Schisandrin B groups,the cell volume was decreased,the intercellular adhesion was disappeared,and the cell membrane was intact but more permeable;the cytoplasm shrank and vacuolar structures appeared inside the cells,with some fragmented and floating on the surface of the solution;in 10.0 mg·L-1 Schisandrin B group,the Pan02 cells exhibited notable apoptotic bodies,indicating an apoptotic state.The EdU staining results showed that compared with control group,the rates of EdU positive cells in 2.5,5.0,and 10.0 mg·L-1 Schisandrin B groups were significantly decreased(P<0.01).The flow cytometry results showed that compared with control group,the percentages of the cells at S phase in 2.5,5.0,and 10.0 mg·L-1 Schisandrin B groups were significantly increased(P<0.01),while the percentages of the cells at G2/M phase were significantly decreased(P<0.01),and the percentages of the cells at G0/G1 phase in 5.0 amd 1.0 mg·L-1 Schisandrin groups were decreased(P<0.01);compared with control group,the apoptotic rates of the cells in 2.5,5.0,and 10.0 mg·L-1 Schisandrin B groups were significantly increased(P<0.01).The Western blotting results showed that compared with control group,the expression levels of p27,B-cell lymphoma 2(Bcl-2)associated X protein(Bax),cleaved cysteine aspartic acid protease-3(cleaved Caspase-3),and cleaved poly adenosine diphosphate(ADP)ribose polymerase(cleaved PARP)proteins in the cells in 2.5 mg·L-1 Schisandrin B group were significantly increased(P<0.05 or P<0.01),the expression levels of cyclin A2,cyclin E2,and Bcl-2 proteins in the cells in 5.0 and 10.0 mg·L-1 Schisandrin B groups were significantly decreased(P<0.05 or P<0.01),while the expression levels of p27,Bax,cleaved Caspase-3,and cleaved PARP proteins in the cells in 5.0 and 10.0 mg·L-1 Schisandrin B groups were significantly increased(P<0.01).Conclusion:Schisandrin B has an inhibitory effect on proliferation of the pancreatic cancer Pan02 cells,and its mechanism may be related to the activation of the cysteine aspartic acid protease-3(Caspase-3)pathway to induce the apoptosis and activating p27 protein to induce the arrest of cell cycle at S phase.

Bioremediation is considered as a cost-effective, efficient and free-of-secondary-pollution technology for petroleum pollution remediation. Due to the limitation of soil environmental conditions and the nature of petroleum pollutants, the insufficient number and the low growth rate of indigenous petroleum-degrading microorganisms in soil lead to long remediation cycle and poor remediation efficiency. Bioaugmentation can effectively improve the biodegradation efficiency. By supplying functional microbes or microbial consortia, immobilized microbes, surfactants and growth substrates, the remediation effect of indigenous microorganisms on petroleum pollutants in soil can be boosted. This article summarizes the reported petroleum-degrading microbes and the main factors influencing microbial remediation of petroleum contaminated soil. Moreover, this article discusses a variety of effective strategies to enhance the bioremediation efficiency, as well as future directions of bioaugmentation strategies.
Biodegradation, Environmental ; Petroleum ; Soil ; Soil Microbiology ; Soil Pollutants

Objective To evaluate feasibility of flap fenestration in management of the condition that a stent-graft is wrongly deployed in false lumen of aortic dissection.Methods A retrospective analysis was conducted on a case of Stanford type B aortic dissection who was confirmed that the stent-graft had been wrongly deployed in false lumen of aortic dissection by CT angiography (CTA) after endovascular repair.Thereafter,an additional surgical management was carried out.At length,flap fenestration at 2 cm below pre-deployed stent-graft was performed using an Outback catheter so as to link false and true lumens.Successively,another stent-graft was placed via the access to draw blood flow from false lumen back to true lumen and then coil embolization was performed for distal breach of the dissection.Results Surgery was carried out under local anesthesia,with operation duration of 170 minutes and intraoperative blood loss of 350 ml.The chest and back pain disappeared after operation.The patient commenced to take food and take off-bed activity at the very day of operation in absence of paraplegia,visceral ischemia,or other complications.Aortic CTA at postoperative one week showed ideal positioning of stent-graft,fluent blood flow of aorta and major visceral artery,and vanishing of false lumen.Conclusions Flap fenestration at far-end of pre-deployed stent-graft so as to draw blood flow back to true lumen by connecting the new placed stent graft to pre-deployed stent-graft is an effective treatment for mis-deployment of stentgraft in false lumen of aortic dissection.Outback catheter for flap fenestration is characterized by accurate location and high safety.