Objective To investigate the effects of exosome(Exo)secreted by cholangiocarcinoma(CCA)cells under hypoxic conditions on the proliferation,migration,invasion,and Janus kinase 2(JAK2)/signal transducer and activator of transcription 3(STAT3)pathway of CCA cells.Methods Exo secreted by CCA cells was extracted and identified.Exo under normoxic and hypoxic conditions were co-cultured with CCA cells and divided into normal oxygen group,hypoxic group,hypoxic+inhibitor group,and blank group.The proliferation,migration,and invasion abilities of CCA cells were tested separately.Immunoblotting was applied to detect the expression levels of relevant proteins.Results Exo carrying fluorescent dye 3,3'-dioctadecyloxacarbocyanine perchlorate was detected in CCA cytoplasm.Compared with blank group,the number of clone formation,cell migration and invasion cells in normoxic group increased,the expression of proliferating cell nuclear antigen(PCNA),matrix metallopeptidase(MMP)-2,MMP-9,neural cadherin(N-Cadherin),Vimentin,phosphorylated JAK2(p-JAK2)/JAK2,phosphorylated STAT3(p-STAT3)/STAT3 increased,and the expression of epithelial cadherin(E-Cadherin)decreased(P＜0.05);Compared with normal oxygen group,the number of clone formation,cell migration and invasion cells in hypoxia group increased,the expression of PCNA,MMP-2,MMP-9,N-Cadherin,Vimentin,p-JAK2/JAK2 and p-STAT3/STAT3 increased,and the expression of E-Cadherin decreased(P＜0.05).Compared with hypoxia group,the number of clone formation,cell migration and invasion cells in hypoxia+inhibitor group decreased,the expression of PCNA,MMP-2,MMP-9,N-Cadherin,Vimentin,p-JAK2/JAK2 and p-STAT3/STAT3 decreased,and the expression of E-Cadherin increased(P＜0.05).Conclusion Exo secreted by CCA cells under hypoxic conditions promote proliferation,migration,invasion,and epithelial mesenchymal transition of CCA cell by activating the JAK2/STAT3 pathway.

Objective To investigate the effects of exosome(Exo)secreted by cholangiocarcinoma(CCA)cells under hypoxic conditions on the proliferation,migration,invasion,and Janus kinase 2(JAK2)/signal transducer and activator of transcription 3(STAT3)pathway of CCA cells.Methods Exo secreted by CCA cells was extracted and identified.Exo under normoxic and hypoxic conditions were co-cultured with CCA cells and divided into normal oxygen group,hypoxic group,hypoxic+inhibitor group,and blank group.The proliferation,migration,and invasion abilities of CCA cells were tested separately.Immunoblotting was applied to detect the expression levels of relevant proteins.Results Exo carrying fluorescent dye 3,3'-dioctadecyloxacarbocyanine perchlorate was detected in CCA cytoplasm.Compared with blank group,the number of clone formation,cell migration and invasion cells in normoxic group increased,the expression of proliferating cell nuclear antigen(PCNA),matrix metallopeptidase(MMP)-2,MMP-9,neural cadherin(N-Cadherin),Vimentin,phosphorylated JAK2(p-JAK2)/JAK2,phosphorylated STAT3(p-STAT3)/STAT3 increased,and the expression of epithelial cadherin(E-Cadherin)decreased(P＜0.05);Compared with normal oxygen group,the number of clone formation,cell migration and invasion cells in hypoxia group increased,the expression of PCNA,MMP-2,MMP-9,N-Cadherin,Vimentin,p-JAK2/JAK2 and p-STAT3/STAT3 increased,and the expression of E-Cadherin decreased(P＜0.05).Compared with hypoxia group,the number of clone formation,cell migration and invasion cells in hypoxia+inhibitor group decreased,the expression of PCNA,MMP-2,MMP-9,N-Cadherin,Vimentin,p-JAK2/JAK2 and p-STAT3/STAT3 decreased,and the expression of E-Cadherin increased(P＜0.05).Conclusion Exo secreted by CCA cells under hypoxic conditions promote proliferation,migration,invasion,and epithelial mesenchymal transition of CCA cell by activating the JAK2/STAT3 pathway.

Fear memories are temporarily suppressed after repeated retrieval, a phenomenon known as memory extinction.How to reduce or even eliminate fear memory is the key to the treatment of fear related diseases such as post-traumatic stress disorder(PTSD). A single extinction training based on Pavlov's fear regulation task could only inhibit the expression of conditioned fear memory traces, but it could not eliminate the acquired conditioned fear memory. However, according to the reconsolidation theory based on memory, the retrieval-extinction paradigm has a more lasting effect on the erasure and rewriting of fear memory, and can effectively prevent the return of fear memory. Studies have shown that extraction-regression is closely related to a variety of neurotransmitter receptors such as glutamate receptor(GluR), dopamine receptor(DAR), L-type voltage-gated calcium channels(LVGCs) and cannabinoid. Moreover, its effect is closely related with factors such as retrieval-extinction memory stage. At present, most of the researches on extracted boundary conditions only stay at the level of behavior, with little understanding and exploration on the level of molecular mechanism. From the perspective of molecular neurobiology, with different stages of memory and different types of receptors and molecular mechanisms, this research reviewed the mechanisms of retrieval-extinction in recent years.It provided valuable signaling pathways, molecular targets and research directions for the treatment of fear-related diseases such as PTSD.

Objective To investigate the effects of application of gauze packing oppression in severe liver trauma therapy.Methods Clinical data of gauze packing oppression treated 18 patients with severe liver laceration were retrospectively analysized,with gauze packing oppression,postoperative hemostatic,antibiotic therapy and nutritional support,a t 5 -7 d began plucking gauze,12 -14 d after pulling,no bleeding wounds gradually healed. Results 17 cases were cured,1 died,the cure rate of 95%,the cause of death as multiple injuries caused by the merger of multiple organ failure;postoperative pull gauze(or bandage)again bleeding in 6 cases,2 cases of secondary hemorrhage,the drug was difficult to control again laparotomy to stop bleeding;after 4 cases of subphrenic effusion, infection,complicated with biliary fistula in 5 cases,liver abscess in 2 cases,3 cases of abdominal infection,wound infection in 3 cases.Conclusion For patients with severe liver rupture gauze packing to stop bleeding is still simple and effective ways to deal under an emergency situation for the hospital,both a method of treatment,but also packing oppression to stop bleeding temporarily for processing and then sent to a higher level hospital completely win time,can effectively reduce the mortality and reduce complications.

Objective To investigate the effects of chronic ethanol exposure and withdrawal on the expression of actin-binding protein cofilin,p-cofilin and cyclin-dependent kinase-5 (cdk5) in the nucleus accumbens and striatum in rat brain.Methods Twenty-four male SD rats were randomly divided into one control group and three experimental groups.In the experimental groups,ethanol was administered in drinking water at the concentration of 6％ (V/V) for two months.Rats in control group drank normal drinking water.After two months ethanol was removed and ethanol withdrawal syndromes were evaluated.Rats were sacrificed on withdrawal 0 h,withdrawal 6 h and withdrawal 2 d.The expression levels of cofilin,p-cofilin(ser3)and cdk5 in the rat brain were measured by immunohistochemistry methods.Results Withdrawal syndrome scores of ethanol fed rats were obviously higher than those of control rats after ethanol was removed,the highest score occurred at 6 h after ethanol withdrawal.In the nucleus accumbens area of rat brain,the levels of cofilin on withdrawal 0 h significantly decreased compared with control group ((0.31±0.05),(0.39± 0.05),P＜ 0.05).The levels of cdk5 on withdrawal 0 h and withdrawal 6 h significantly increased compared with control group((0.36±0.07),(0.34±0.07),(0.25±0.05),P＜0.05).In the striatum of rat brain,the levels of cofilin on withdrawal 0 h significantly decreased compared with control group ((0.26±0.04),(0.34±0.05),P＜0.05).The levels of p-cofilin on withdrawal 6 h significantly increased compared with control group((0.43±0.06),(0.30±0.06),P＜0.01).The levels of cdk5 on withdrawal 0 h significantly increased compared with control group((0.35±0.06),(0.26±0.05),P＜0.05),and the levels of cofilin on withdrawal 6 h significantly decreased compared with control group((0.37±0.06),(0.26±0.05),P＜0.01).Conclusion Chronic ethanol exposure can induce the development of ethanol dependence,and it accompanies with changes in the expression of actin-binding protein and cdk5 in the brain of rats.