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Objective To explore the effect of toll-like receptor 4 (TLR4), myeloid differentiation protein-2 (MD2), and stromal interaction molecular 1 (STIM1) for regulating human vascular endothelial calcium overload injury and inflammatory reaction induced by bacterial endotoxin (LPS).Methods Human umbilical vein endothelial cells (HUVECs) were cultured in Dulbecco's modification of Eagle's medium (DMEM). ① The levels of TLR4, MD2 and nuclear factor-κB (NF-κB) were detected by reverse transcriotion-polymerase chain reaction (RT-PCR) before and 0.5, 1, 6, 12, 24 hours after LPS stimulation. ② Intracellular calcium peak level was detected by confocal following probe fluo-3 AM loading in HUVEC cells induced with LPS and transfected by psiSTIM or psiTLR. ③ MD2, STIM1 or NF-κB protein level was detected by immunoprecipitation (IP) and immuno-blotting in HUVEC cells which were transfected by TLR4 inhibited expression (psiTLR) for 12 hours and followed by LPS stimulation for 6 hours. ④ HUVEC cells were randomly divided into 6 groups: control group, LPS group, PDTC 0.1 mg/L group, PDTC 1 mg/L group, psiTLR 1 h group and psiTLR 12 h group. Tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) were detected by enzyme linked immunosorbent assay (ELISA) in supernatant. The mRNA levels of STIM1 and NF-κB were detected by RT-PCR.Results ① The mRNA levels of TLR4, MD2, and NF-κB gradually increased after LPS induction and peaked at 6 hours (2-ΔΔCt: 23.52±2.88, 17.43±3.43, 18.13±2.99, respectively), which were statistically significant before the stimulation with LPS (2-ΔΔCt: 7.02±2.81, 5.19±3.22, 8.11±1.42, allP < 0.05). ② Extracellular calcium influx in LPS group was increased significantly higher than control group (nmol/L: 108.13±22.33 vs. 41.57±13.19, P < 0.01). Extracellular calcium influx in psiSTIM+LPS group (nmol/L: 62.61±14.12 vs. 108.13±22.33,P < 0.05) and psiTLR+LPS group (nmol/L: 50.78±8.05 vs. 109.43±20.21,P < 0.01) were both suppressed as compared with LPS group. While extracellular calcium peak level in psiTLR+psiSTIM+LPS group further decreased (nmol/L: 39.31±6.42 vs. 109.43±20.21,P < 0.01). ③ MD2 protein but not STIM1 or NF-κB can be detected in anti-TLR4 precipitates in control (ctrl-) by immunoprecipitation. MD2 protein level increased in anti-TLR4 precipitates in LPS group (ctrl+) and was suppressed in TLR4 inhibiting group (psiTLR). ④ The levels of TNF-α in PDTC 1mg/L group were significantly lower than those of LPS group (ng/L: 0.60±0.24 vs. 1.77±0.66,P < 0.01). The levels of IL-6 in PDTC 0.1 mg/L, 1 mg/L group and psiTLR 12 h group decreased significantly lower than that of LPS group (ng/L: 232.10±63.54, 134.32±37.23, 284.23±56.14 vs. 510.22±89.23, allP < 0.05). Compared to LPS group, the mRNA levels of NF-κB and STIM1 were obviously inhibited in PDTC1 mg/L group and psiTLR 12 h group [NF-κB mRNA (2-ΔΔCt): 17.22±2.35, 13.24±3.54 vs. 30.16±2.06; STIM1 mRNA (2-ΔΔCt): 12.57±2.43, 12.21±2.46 vs. 25.12±2.02, allP < 0.05]. Conclusions TLR4, MD2, NF-κB signal and SOC calcium channel STIM1 mediate LPS induced-calcium influx and inflammatory mediators level in HUVEC cells. Extracellular calcium overload and inflammatory response by endotoxin induction can be effectively inhibited by down-regulation of TLR4, NF-κB and/or STIM1.
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Objective To investigate the classification of cerebral arteriovenous malformation (AVM) associated with hemodynamics correlative aneurysms and its efficiency treated by endovascular treatment.Methods The clinical data of 17 patients of AVM associated with hemodynamics correlative aneurysms undergoing endovascular treatment were analyzed retrospectively.Results Eleven cases of aneurysms with great divergence between the aneurysm and arteriovenous malformation were embolized,6 cases of aneurysms without a great divergence between the aneurysm and arteriovenous malformation,1 case of aneurysm was treated with stent,other 5 cases of aneurysms were not treated.Eleven cases of arteriovenous malformations were embolized completely,4 cases were embolized 71％-90％ and 2 cases were embolized 50％-70％.Six cases with residual were given radiotherapy.Follow-up 3 months to 3 years,there were no cases of cerebral hemorrhage or death.Unhandled 5 cases of aneurysm and 1 case of stent implantation with the follow-up by using digital subtraction angiography,laneurysm with a stent was closed.Three aneurysms were disappeared and 2 aneurysms were reduced significantly among the 5 cases of aneurysms without treatment.One case of aneurysm occlusion in patients with stent implantation.Conclusions Classification based on a great divergence artery or not between the aneurysm and arteriovenous malformation is more instructive for clinical treatmen of cerebral arteriovenous malformation associated with hemodynamics correlative aneurysm.If it has not a great divergence artery between aneurysm and arteriovenous malformation,arteriovenous malformation after a thorough treatment,aneurysms need not be treated.The endovascular treatment for cerebral arteriovenous malformation associated with hemodynamics correlative aneurysms has a good efficiency and can be treated as a priority.
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Objective To assess the feasibility and safety of combined transgastric and transcolonic endoscopic cholecystectomy in porcine model.Methods Five female miniature swines were subjected to cholecystectomy by the combined transgastric and transcolonic approach using two flexible endoscopes.The gastric wall of the swine was punctured with a needle knife,followed by dilatation with a balloon-dilator via the transgastric access.then a double channel endoscope was advanced into the peritoneal cavity.Another double-channel endoscope was advanced via a transcolonic access.Gallbladder excision was performed using transcolonic endoscope assisted by transgastric endoscope.After eholecystectomy the gastric and the colonic incisions were closed with clips.The animal was examined 14 days after operation.Results Cholecystectomy was failed in 4 swines because of incomplete exposing the gallbladder(3 swines)and hemarrhage during separating the gallbladder from the liver bed(1 swine).Only one swine was successfully performed cholecystectomy.But 3 perforations were found in the diaphragm which might be done by transcolonic endoscope.The swine lost 4 kg 14 days after the operation.The necropsy revealed a complete transmural healing of the gastric incision with minimal adhesions and a complete healing of the colonic incision with heavy adhesions.An abscess with severe adhesions was found in the right lower quadrant.One perforation of the diaphragm healed,and the other two perfolrations merged and formed a diverticulum embedded with the liver.Conclusion Combined transgastric and transcolonic approach appears difficult for cholecystectomy and the safety remains tO be further studied.
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Objective To investigate the feasibility and safety of trans-gastric route combined with trans-colonic or trans-vaginal route endoscopic salpingectomy and liver biopsy, as methods of natural orifice translumenal endoscopic surgery (NOTES). Methods Endoscopic salpingectomy and liver biopsy were per-formed in 4 female porcine models, in 2 of which the procedure was achieved via trans-gastric and trans-co-Ionic route (Group A), while in other 2 was via trans-gastric and trans-vaginal route (Group B). The ani-mals were sacrificed after the procedure and the related complications were observed. Results Abdominal cavity exploration, salpingectomy and liver biopsy could be performed successfully through two combined routes. Perforation of adjacent colon was found in 1 animal with the trans-colonic route NOTES. Conclusion NOTES via trans-gastric and trans-colonic routes or via trans-gastric and trans-vaginal for salpingectomy and liver biopsy are both feasible, while trans-vaginal route seems to be safer than trans-colunic route.
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Objective To investigate the causes of success or failure of natural orifice translumenal endoscopic surgery(NOTES)in animals.Methods Data were collected from animal studies of NOTES carried out between August 2007 and September 2009.The studies were divided into acute phase and survival phase.In the studies,simple NOTES and laparoscopic-assisted NOTES were performed on female miniswines or adult mongrel dogs.Diagnostic abdominal exploration,liver biopsy,cholecystectomy,tubal ligation,oophorectomy,gastrojejunal anastomosis,and splenectomy were made in the animals.The intra-and post-operative complications and technical difficulties were recorded during the procedures.Results Totally 37 miniswines and 16 mongrel dogs were enrolled in the studies of NOTES procedures(91 procedures),including 22 acute experiments and 31 survival experiments.Simple NOTES was conducted through the transgastric route in 25 cases,transcolonic route in 8 cases,and combinations of transgastric and transcolonic routes in 16 cases,and combinations of transgastric and transvaginal routes in 6.In Laparoscopic-assisted cases,36 animals were treated via the transgastric route by traditional(26 cases)or single-port transumbilical laparoscopy(10 cases).Diagnostic exploration was carried out in 33 cases,liver biopsy was done in 11 cases,tubal ligation was made in 11,and oophorectomy was performed on 6;the procedures were all completed in these cases showing a success rate of 100%.Cholecystectomy was completed in 27 cases with a success rate of 44.4%(12/27).One case of splenectomy,and two cases of gastrojejunal anastomosis failed.The totally complication rate in the studies were 39.6%(21/53).The complications included perforation(8 cases),organ injury(5 cases),bleeding(4 cases),infection(3 cases)and pneumoperitoneum with hypertonia(1 case).Conclusions NOTES is feasible,safe and effective in diagnostic abdominal exploration and simple abdominal surgeries,but does not suit difficult abdominal surgeries.By using laparoscopy,the technique may be further developed and used in the future.