Acute pancreatitis (AP) is a severe disease with an increasing incidence rate in clinical practice. Although most patients have mild pancreatitis, the fatality rate of severe pancreatitis remains at a relatively high level, and therefore, early-stage, simple, and accurate clinical scoring systems are urgently needed to determine the severity of AP, so as to facilitate effective disease management and symptomatic treatment and reduce the fatality rate of patients. At present, a large number of studies have demonstrated that the scoring systems such as Ranson score, APACHE Ⅱ score, BISAP score, CTSI score, and some serological markers have been used to evaluate the severity and prognosis of AP, but all of them have certain limitations. This article reviews the research advances in the existing scoring systems, single serological markers, and related modified scoring systems in recent years. Through a literature review, it is concluded that there is no a single scoring system or a single indicator that can cover the whole process of AP diagnosis and treatment and accurately judge the severity of AP, and therefore, it is necessary to develop a new scoring system or combine various indicators for comprehensive evaluation.

OBJECTIVE：To provid e reference for the medicinal resources and clinical application of Tibetan medicine “Dida”. METHODS：Delphi method was adopted. By reviewing literatures ，confirming consultation scope ，inviting experts engaged in clinical，scientific research ，teaching and production of Tibetan medicine. Two methods ，online inquiry and on-site questionnaire ， were used for expert consultation to evaluate the differences and problems existing in the utilization of “Dida”medicinal resources until a consensus was reached ，consensus on the medicinal resources and clinical application of Tibetan medicine “Dida”was determined finally. RESULTS & CONCLUSIONS ：A total of 33 experts participated in the two rounds of consultation. According to the results of literature research ，the first round set up 16 inquiry indicators ；and then according to expert opinions to modify the index system ，the second round set up 18 inquiry indicators. After two rounds of inquiry ，a consensus was finally reached on 16 items on the original name ，quality standards and clinical application of Tibetan medicine “Dida”，and 2 items related to the characteristics and compatibilities of “Dida”had not reach common views. The consensuses of 16 items mainly cover the original name of “Dida”and the evolution of geographic information ，the rational selection of “Dida”medicinal materials ，the effectiveness and safety of clinical use of “Dida”.

We have previously reported that Cystatin C (CysC) is a pivotal mediator in the neuroprotection induced by hyperbaric oxygen (HBO) preconditioning; however, the underlying mechanism and how CysC changes after stroke are not clear. In the present study, we demonstrated that CysC expression was elevated as early as 3 h after reperfusion, and this was further enhanced by HBO preconditioning. Concurrently, LC3-II and Beclin-1, two positive-markers for autophagy induction, exhibited increases similar to CysC, while knockdown of CysC blocked these elevations. As a marker of autophagy inhibition, p62 was downregulated by HBO preconditioning and this was blocked by CysC knockdown. Besides, the beneficial effects of preserving lysosomal membrane integrity and enhancing autolysosome formation induced by HBO preconditioning were abolished in CysC rats. Furthermore, we demonstrated that exogenous CysC reduced the neurological deficits and infarct volume after brain ischemic injury, while 3-methyladenine partially reversed this neuroprotection. In the present study, we showed that CysC is biochemically and morphologically essential for promoting autophagic flux, and highlighted the translational potential of HBO preconditioning and CysC for stroke treatment.
Animals ; Autophagy ; physiology ; Beclin-1 ; metabolism ; Brain ; metabolism ; pathology ; Brain Ischemia ; metabolism ; pathology ; therapy ; Cystatin C ; genetics ; metabolism ; Disease Models, Animal ; Gene Expression ; Gene Knockdown Techniques ; Hyperbaric Oxygenation ; Lysosomes ; metabolism ; pathology ; Male ; Microtubule-Associated Proteins ; metabolism ; Neurons ; metabolism ; pathology ; Neuroprotection ; physiology ; Oxygen ; therapeutic use ; Random Allocation ; Rats, Sprague-Dawley ; Rats, Transgenic ; Reperfusion Injury ; metabolism ; pathology ; therapy

Stroke is a leading cause of death worldwide. Up to one thousand potential drugs or interventions have been developed to treat stroke, out of which ~160 have gone on to clinical trials. However, none of them has been successful. New insights into the molecular and cellular mechanisms of ischemia-induced injury are needed for discovering new therapeutic targets. Recently, Drosophila has been used to uncover new hypoxia-related genes. In this study, we describe an efficient and reliable assay with a sophisticated apparatus for studying the effects of oxygen deprivation on flies. Using this assay, wild-type flies were exposed to an anoxic environment for varying lengths of time, then the cumulative death rate and mobility recovery were systematically analyzed. We found that anoxia for over one hour caused lethality. The cumulative death rate on day 5 after anoxia was linearly and positively correlated with the duration of anoxia, and reached 50% when the duration was 2.5 h-3 h. We also found that the mobility recovery in normoxia was slow, as the climbing ability remained largely unchanged 4 h-6 h after 2.5-h of anoxia. We suggest that 2.5 h-3 h of anoxia and 4 h-6 h of recovery before mobility analysis are appropriate for future use of the anoxia assay.
Animals ; Behavior, Animal ; Disease Models, Animal ; Drosophila melanogaster ; Hypoxia

[Abstract] Objective: To investigate the effect of sphingosine kinase 1 (SphK1) knockdown on the proliferation and migration of colon cancer RKO cells induced by mesenchymal stem cells (MSCs). Methods: RKO cells were treated with MSCs conditioned medium (MSC-CM) or control medium (Control-CM), respectively. Cell proliferation was detected by CCK-8 assay. Cell migration ability was tested by Transwell chamber assay. The proteins expression of Ki-67, MMP-2/9, CD44 and CD133 was detected by Western blotting. Then, the expression of SphK1 in RKO cells was suppressed by targeted gene lentivirus shRNA vector transfection. The effects of SphK1 knockdown on the proliferation, migration and protein expressions of Ki-67, MMP-2/9, CD44 and CD133 of RKO cells induced by MSC-CM were observed. Results: The RKO cells proliferation was promoted by MSC-CM in a time-dependent manner; moreover (P<0.05), the migration ability of cells was significantly enhanced after being treated with MSC-CM(P<0.01). In addition, MSC-CM significantly increased the protein expressions of Ki-67, MMP-2/9, CD44 and CD133(all P<0.05 or P<0.01). Lentiviral ShRNA vector transfection could significantly inhibit the expression of SphK1. Down-regulation of SphK1 significantly inhibited the proliferation, migration and protein expressions of Ki-67, MMP-2/9, CD44 and CD133 of RKO cells induced by MSC-CM(all P<0.05 or P<0.01). Conclusion: MSC-CM promotes the proliferation and migration of colon cancer RKO cells. Down-regulation of SphK1 reverses the cell proliferation and migration induced by MSC-CM via inhibiting the expression of MMP-2/9, CD44 and CD133.

[ ABSTRACT] AIM:To investigate the effects of sphingosine kinase l ( SphK1) and focal adhesion kinase ( FAK) on the epithelial-mesenchymal transition ( EMT) of human colon cancer HCT 116 cells.METHODS:Human colon cancer HCT116 cells were divided into 3 groups.N, N-dimethylsphingosine (DMS) was used to suppress the activity of SphK1. PF573228 was used to suppress the activation of FAK .The cells treated with equal volume of culture medium severed as control group.The cell viability was measured by MTT assay .The protein expression of SphK1, FAK and the EMT relative protein E-cadherin, N-cadherin, vimentin and matrix metalloproteinase (MMP) 2 was analyzed by Western blot.The mR-NA expression of SphK1, sphingosine-1-phosphate (S1P), FAK, E-cadherin and vimentin was detected by real-time PCR. The ability of tumor cell migration was measured by wound-healing assay.RESULTS:The cell viability of HCT116 cells was suppressed by DMS and PF 573228 in dose and time dependent manners .DMS significantly suppressed the expression of SphK1, FAK, N-cadherin, vimentin and MMP2, meanwhile enhanced the expression of E-cadherin.PF573228 reduced the expression of FAK , SphK1, N-cadherin, vimentin and MMP2, meanwhile increased the expression of E-cadherin (P<0.01).In addition, the migration ability of HCT116 cells was significantly decreased by treating with DMS and PF573228 (P<0.01).Compared with control group , the mRNA expression of FAK, SphK1, S1P and vimentin was de-creased, while the expression of E-cadherin was increased significantly in PF573228 group and DMS group (P<0.05). CONCLUSION:SphK1 and FAK signaling pathways may play an important role in the occurrence of EMT in the colon cancer HCT116 cells.

Objective To evaluate the therapeutic effect of endoscopic management of plastic stents of post-liver transplantation anastomotic biliary stricture. Methods From January 2010 to October 2015, clinical data of patients with post-liver transplantation anastomotic biliary stricture and received endoscopic retrograde cholangiopancreatog﹣raphy and plastic stents management was collected. The technical success rate, ERCP-related complications, clinical remission rate and long-term complications were main outcome measurements to compare the efficacy and safety of different number of stents in managing post-liver transplantation anastomotic biliary stricture. Results Among the 18 patients (0.5 ~ 60.0 months) with post-liver transplantation ABSs, seven patients received less plastic stents treat﹣ment (< 3 stents), nine patients with persistent anastomotic or recurrent stricture received multiple plastic stents treatment (≥ 3 stents), two patients received multiple plastic stents treatment once suffered with post-liver trans﹣plantation ABSs. The endoscopic technical success rate was seventy-six over eighty (95.0 %). Among the seven pa﹣tients received less plastic stents treatment, one loss to follow-up, two were still under treatment, one died of acute hepatic failure, one died of septic stock, one combined with biliary fistula resulted in treatment failure, one achieved clinical remission, the clinical remission rate was one third (33.3 %). Among the eleven patients received multiple plastic stents treatment, two loss to follow-up, one was still under treatment, two received surgery because of failed treatment, six achieved clinical remission, the clinical remission rate was 75.0 % (6/8). The average diameters and stent durations of management of 1 stent, 2 stents, 3 stents, 4 stents, 5 stents, 7 stents were 8.5 F, 17.0 F, 24.0 F, 28.0 F, 36.0 F, 50.0 F. Among the six early postoperative complications, five cases occurred in less stent manage﹣ment and one occurred in MPSs management, the early postoperative complication rate was 7.5 %(6/80). No severe ERCP-related complications and procedure-related deaths. Conclusions Endoscopic management of plastic stents is safe and effective for post-liver transplantation ABSs. Providing larger biliary support, the multiple plastic stents treatment was superior to less plastic stents treatment in view of clinical remission rate, especially for refractory one. Multiple plastic stents did not increase the incidence of complications, it could be used as the first-line treatment of post-liver transplantation duct-to-duct biliary anastomosis for its safety and effectivity.

Objective To investigate the correlation between sperm-nucleporotein transition and sperm parameters and embryo development,also to evaluate the influence of pregnancy out comes of assisted reproductive technology (ART).Methods Sperm-nucleoprotein transition assay of a total of 676 patients underwent ART treatment were detected by aniline blue staining,and the correlation analysis between spermnucleoprotein transition and sperm parameters,DNA damage,acrosin activity,fertilization rate,cleavage rate,quality of early embryo development as well as blastocyst formation rate was performed.Results The sperm concentration,(a+b) ％ sperm,sperm count and acrosin activity was (66.5±4.6) × 109/L,(149.2±9.9)×109/L,(51.2±1.3)％ and (72.2±3.3) mU/106 sperm in abnormal group,and (91.9±2.7) ×109/L,(240.0±8.0) ×109/L,(57.3±0.8)％ and (85.7±1.9) mU/106 sperm in normal group,which reached significant difference (P＜0.01).DNA fragmentation index (DFI) was (17.3± 1.0)％ in abnormal group,which was significantly higher than (14.6±0.5)％ in normal group.The cleavage rate of 95.0％,D3/D5 high quality embryo rates of 34.2％ and 1.28％,D5 blastocyst formation rate and the total rate of blastocyst formation rate of 22.4％ and 38.6％ in abnormal group,which were significantly lower than that in normal group (96.9％,38.2％,2.70％,27.9％ and 46.4％) (P＜0.01).The rate of spontaneous abortion was 12.3％ in abnormal group,which was significantly higher than that in normal group (4.7％) (P＜0.01).However,there was no significant difference in biochemical pregnancy rate and ectopic pregnancy rate between the 2 groups (P＞0.05).Conclusions Sperm-nucleoprotein transition was positively related with sperm parameters,DNA damage,acrosin activity,and also has an adverse effect on embryo development and the outcomes of ART.It is suggested that the sperm-nucleoprotein transition should be detected before ART.

Objective To investigate the expression of sphingosine kinase 1(SphK1)and NF-κB in colon carcinoma tissues and their correlation with clinicopathologic features.Methods Sixty-six paraffinembedded colon carcinoma samples and 66 fresh colon carcinoma samples were tested using immunohistochemistry,RT-PCR and Western blot,respectively.Results In 66 fresh colon carcinoma samples,the positive rate of SphK1 and NF-κB mRNA expression were 84.85％(56/66)and 74.24％(49/66),while the positive rate of SphK1 and NF-κB protein detected by Western blot were 78.79％(52/66)and 69.70％(46/66).The positive rates were higher than those in the adjacent tissues[mRNA:63.64％(42/66),48.49％(32/66);protein:57.58％(38/66),45.45％(30/66)]and the normal mucosa [mRNA:42.42％(28/66),25.76％(17/66); protein:36.36％(24/66),24.24％(16/66)],with statistical significances(all P values ＜ 0.05).The mean expressive levels of SphK1 and NF-kB mRNA and protein in colon carcinoma were both significantly higher than those in the adjacent tissues and the normal mucosa(mRNA:0.55±0.06 vs0.35 ±0.05 vs0.25±0.05,0.75 ±0.06 vs0.43±0.05 vs0.30±0.04 ; protein:0.77 ± 0.05 vs 0.38 ± 0.06 vs 0.12 ± 0.03,0.45 ± 0.08 vs 0.23 ± 0.05 vs 0.13 ± 0.03 ;all P values ＜ 0.05).There was a close correlation between SphK1 and NF-kB expression levels (r =0.459,P =0.036).The results of immunohistochemistry were similar to those of RT-PCR and Western blot.Overexpression of SphK1 and NF-κB in colon carcinoma was related with depth of invasion,distant and lymph node metastasis and Dukes'stages(all P values ＜0.05).The expression of SphK1 was also related with differentiation(P ＜ 0.05).Conclusions Overexpression of SphK1 and NF-κB may be involved in the pathogenesis and progression of colon carcinoma.Moreover,SphK1 and NF-κB may be correlated with the invasion and metastasis of colon carcinoma.