| With the advance of urbanization,environmental pollution of food,water and air has intensified,due to an annuallyincreased morbidity and mortality of gastrointestinal cancer,among them,the morbidity of gastric cancer and colorectal cancer is especially high.For early and mid-term gastric and colorectal cancer patients without surgical contraindications,surgical removal is the first choice of treatment;and for the advanced,the main strategy is chemotherapy,targeted therapy.Because of the development of anastomosis consumables as gastrointestinal staplers and hemostatic and tissue separating equipmentas ultrasonic scalpel,surgeons havemore convenient and safe choices for gastrointestinal anastomosis and tissue separation.With the development of medical technology and devices,the new gastrointestinal stapling technology,products and hemostatic equipments have been upgraded,to providedgastrointestinal surgeons more convenient and effective surgical instruments,greatly shorten the operation time,reduce the risk of surgery on patients;however,surgeonswill never stop to seek more efficient,safe and convenient method of gastrointestinal anastomosis and vascular closure.High-frequency electric welding(HFEW)on soft tissue in vivo is a new field of electrosurgery in recent years.This technology is the latest technology researched and developed by the Ukrainian National Academy of Sciences cooperated with"E.O.Paton Electric Welding Institute",its technologicalapplication form is anEKVZ-300PATONMED high-frequency electronic welding equipment.It is a bipolar electrosurgical equipment which has a closed loopautomatic control systemtechnology,accurately heating human soft tissues and organs woundsto connectby high-frequency current.When heating soft tissue by high-frequency current,which proteinswill be denatured,the thermal molecular of which accelerated,so that space and form a plurality of joints in endosperm at the bonding interface,the cooling process is accompanied with protein denaturation and refolding,it can make sure the minimized damage and cell death of surrounding tissue when occluding the blood vessels or other tissues,and prevent rupture of anastomotic ends to ensure the safety and efficiency of the blood vessel disconnection.It replaces the surgical technique involves the application of standard surgical tools.According to instruction ofthe instrument,coagulation mode can be used to close vessels and separate tissue,bond(welding)model can be used to anastomose archentericorgans;adhesion(welding)model is set into 10 grades:dA0,dAI,dA2,dA3,dA4,dA5,dA6,dA7,dA8,dA9.In the operation of stomach andintestines,effective transection,of vessels and separation of tissues is the key technique.Ligationof small vessels and separation of tissues has developed from the early application of silk thread into a variety of electrical surgical equipment or ultrasonic equipment;due to consuming time,demanding large space,and difficult to operate,silk suture in laparoscopic surgery is low efficiency,and has been replaced by and other surgical equipments,among which,ultrasonic knives and LigaSure are used to transection 1-3mm vasculars;and titanium clip and Hem-O-lock ligating usually used to transection vesselsmore than 3mm after clipping.However,on the one hand,common clinical vascular ing devicesas LigaSure,ultrasound knife are not very dependable when deal with large vessels,vesselsmore than 7mmstill can not be ligatured and transected,and titanium clips,Hem-O-lock ligating clips still have the problems as fall-off and rejection reaction;in another hand,common clinical vascular devices costs too long time and makes large injury when ligatingvessels,allof those have prompted new generation of surgical vascular ligation and tissue separation equipments.With the development of laparoscopic gastrointestinal surgery,a fast,safety and stability vascular and tissue separation equipment is more and more urgently required;in addition,there is no data to compare the safety,scope and efficiency of the instruments and equipments for vascular ligation.Gastrointestinal anastomosis is the keytechnique of the abdominal surgery,once there is a complication asanastomotic leakage or bleeding complications,may lead to serious consequences,and even threatenlife.The traditional methods such as gastrointestinal anastomosis stapler,manual suture play an important role in recovery of gastrointestinal system continuity.However,manual suture of gastrointestinal tract conducted by less skilled surgeons will not only spend a lot ofoperating time,but also increase the risk of complicationspostoperative;usingstapler can save time,but also hasdisadvantages of cutting and anastomosis incomplete,tissue incarceration,excitation on unsuitable position,difficult to withdraw,and expensive,so itis limited when applyingon the entire gastrointestinal digestive tract.However,there is no creative development in the field of gastrointestinal anastomosis.At present,gastrointestinal anastomosis stapler and GIAare mainly used to cut and suturesmall intestineboth at home and abroad,but itis expensive and nonreusable.The usage of soft tissue high-frequency welding technique for gastrointestinal anastomosishas not been reported inland.It has been reported abroad that the HFEW-300PATONMED high frequency electronicwelding apparatus,which is a kind of applications of soft tissue high-frequency welding technology,can effectively weld detachedretina of rabbit.Our preliminary study shows that the equipment caneffectively ligate animal(pig)small intestinein vitro,compared with hand suture,intestine ligated with thewelding equipmenthas obvious advantages in aspect of the bursting pressure,operation time,etc,but it has not been reported whether it can effectivelyligate small intestine of animal(pig)in vivo.This study will study whether soft tissue high-frequency welding technology can effectively ligate small intestine of animal(pig)in vivo,and the relevant ligating operation skills and welding mechanism;to provide scientific basis for subsequent clinical trials,to lay the foundation of validation and promotion of soft Tissue High-Frequency welding technology.Patients with advanced gastric cancer as no significant obstructive symptoms,often were given to chemotherapy or targeted therapy。Research shows,Induce metabolic changes in the microenvironment and uncontrolled inflammatory response were the important factors of gastric carcinogenesis and development.Warburg effect was discovered by the German biochemist Otto Warburg(Otto Warburg,1883～1970)in 1930,the way of tumor cells produce energy was very special:the healthy cells rely on mitochondrial oxidative carbohydrate molecules release more energy,but most of the tumor cells to release a small amount of energy in the way of glycolysis for themselves energy supply。In the case of hypoxia,HIF-la activation of a key enzyme in the glycolytic key enzyme upregulation and downregulation of oxidative phosphorylation to enhance the Warburg effect,HIF-1α was the key transcriptional activator which regulatd tumor cells adapt to hypoxia microenvironment.Study confirms that intracellular highly expressed HIF-1αcould promote tumor cell heterogeneity and tumor angiogenesis,led to tumor cell invasion and metastasis.This study demonstrated that RA effectively inhibit the expression of HIF-1α,inhibition Warburg effect of MKN45 cells,promote MKN45 gastric cancer cells transferred.Research shows,STAT3 was over expressed in gastric cancer cells,and related to tumorigenesis.Proinflammatory cytokines such as IL-6,IL-1β,TNF-α were effective STAT3 activator.Cancer survival,proliferation and angiogenesis-related genes are downstream targets of STAT3.This study shows,RA regulate of proinflammatory cytokines such as IL-6,IL-1β,TNF-α by inhibiting the expression of miR-155-5p,which inhibitd the IL-6/STAT3 pathway of cellular suppressing the role of the Warburg effect in MKN45 cells.We demonstrated that,miR-155 is the goal target,which RA acts on Warburg effect by inhibiting IL-6/STAT3 pathway.Section I:Comparison of a new high-frequency electric welding system with other vessel ligation modalities in a rabbit modelObjective:Various hemostatic surgical devices are in current use,but improvements are always desired.Thus,we compared the feasibility,effectiveness,and safety of a new high-frequency electric welding system for blood vessel ligation with other available hemostatic modalities.Methods:Thirty-two New Zealand white rabbits were randomly allocated into four groups.Large,small(1-3 mm)and medium(4-5 mm)blood vessels from the rabbits were ligated with ultrasonic coagulating shears,LigaSure vessel sealing system,a high-frequency electric welding system or tie ligation in vivo.Vessel diameter and bursting pressures were measured in vivo.Thermal damage caused by surgical modalities was evaluated by peripheral energy spread,histopathology,and mRNA expression of iNOS and eNOS from harvested blood vessels.Results:Vessel diameter,seal time,and burst pressures were measured in 98 arteries and 91 veins.The burst pressures of artery/vein from LigaSure,high-frequency electric welding(HFEW)and ultrasonic coagulating shears(HA)was 308.24/327.44 mm Hg,507.03/488.25 mm Hg,and 500.50/477.37 mm Hg,respectively.Compared to Ligasure,HFEW and UCS had greater bursting pressures for sealing small and medium blood vessels(p<0.001).The average temperature within 2 cm of the welding center from LigaSure,HFEW,UCS was 87,99 andl37℃,respectively.The full-thickness peripheral energy spread from LigaSure,HFEW,HA was 1.6,1.9 and 2.5 mm,respectively(p<0.001).HFEW produced the fewest acute histological changes.Inducible and endothelial nitric oxide synthase expression was not observed after application of these methods,compared to the tie ligation group(P>0.01).Conclusion:There may be no clinical limitations for applying various hemostatic devices to small and medium vessels(up to 5 mm)under physiologic blood pressures.However,HFEW offered the fewest acute histological differences,likely due to less thermal injury of whole vessel wall.Therefore,the HFEW is an efficient and safe vessel sealing system.Section Ⅱ:A New equipment of High-Frequency Electric Welding for comparison and evaluation of vessel ligation in vivo procine modelObjective:Compared of the feasibility,effectiveness,and safety of a new high-frequency electric welding system for blood vessel ligation with HA、Ligasure and TL other available hemostatic modalities in vivo procine model。Methods:The vessels of three adult pigs were randomly allocated into four groups.Small(1-3 mm)、medium(4-5 mm)and big(5-11 mm)blood vessels from the pigs were ligated with Ultracision harmonic scalpel(HA),LigaSure(LS)vessel sealing system,a high-frequency electric welding(HFEW)system or tie ligation(TL)in vivo.Vessel diameter and bursting pressures were measured in vivo.Thermal damage caused by surgical modalities was evaluated by Fluke Ti27 Thermal Imager。Results:1.A total of 42 arteries and 82 veins,including abdominal aorta,femoral artery and vein,were studied using different ligation modalities.2.The average seal time of LS,HA,HFEW and TL were 6.00±0.00s,9.94 ± 3.19s,7.45±1.64s and 18.15±3.31s,respectively.3.The vessel diameter of the Ligasure.HFEW、HA and TL were 4.51±2.38mm,4.89±2.87mm,4.62±1.88mm and 5.02±2.28mm。but,no significant difference was noted for vessel diameter among the four ligation modalities,respectively(p>0.05)4.The burst pressures of vessels from LigaSure、HFEW、HA and TL was:490.39±186.83mmHg、534.36±211.93 mmHg、391.50±184.00 mmHg 和602.00±132.15mmHg。5.The burst pressures of artery/vein from LigaSure,high-frequency electric welding(HFEW)and ultrasonic coagulating shears(HA)was 490.39±186.83/467.00±128.84mmHg,534±184.00/455.09±100.66mmHg,and 391 ±125.37/319.70±125.37 mmHg,respectively.6.The burst pressure analysis for vessels with a diameter greater than 5 mm,the burst pressure was statistically higher with HFEW(337.83±179.84mmHg)than by HA(218.26±172.59mmHg)(P>0.05).However,for vessels less than 5mm mean burst pressure caused by HFEW(548.00±179.19mmHg)was statistically significantly higher than HA(416.86±153.33mmHg),but this was not statistically significantly different compared to burst pressure caused by LS(513.95±168.96mmHg)。Conclusion:1.HFEW had shorter time for sealing blood vessels than HA and TL,but longer time than LS groups;2.The burst pressures of vessels form HFEW was more than HA,less than TL,there was no significant differences in mean burst pressures in vessels after HFEW and LigaSure;3.The HFEW,HA,LigaSure and TL can effectively sealed arteries in vivo procine model,The bursting pressure of vessel from the four ligation modalities were higher than normal blood pressure of experimental pigs;4.Compared to HA,HFEW had greater bursting pressures for sealing vein blood vessels,but not between HFEW and LS groups;5.The burst pressures of artery vessels form HFEW was more than HA(P<0.05),less than TL(P<0.05),there was no significant differences in mean burst pressures in artery vessels after HFEW and LigaSure;6.HFEW could seal less than 5mm in vivo vessels(artery and vein),the bursting pressure of vessels were greater than HA(P<0.05),less than TL(P<0.05);7.The three devices used in this study seal small、medium and big vessels safely in vivo pigs model;Section Ⅲ:Comparison of a new high-frequency electric welding system for intestinal closure with hand-sewn in vivo procine modelObjective:Various surgical small intestinal anastomosis method is in current use,but improvements are always desired.Thus,we compared the feasibility,effectiveness,and safety of a new high-frequency electric welding system(HFEW)for sealing small bowel with hand-sewn in vivo porcine model.Methods:The 96 bowel segments of three porcine were randomized to be sutured either by HFEW-300 PATONMED or hand-sewn and mucosa-to-mucosa fusions were subjected in vivo porcine.Bursting pressures,suture time,thermal damage and the temperature of sealed ends were measured.Results:Segments ligature with hand-sutured and double-sealed with HFEW were compared.Burst pressure of hand-sutured group had a significantly higher pressure than the HFEW group(136.19 mmHg vs 75.83 mmHg,P<0.01).All 48 porcine small bowels closed by the HFEW-300 generator showed a success rate of 100%.The closing time of HFEW group(108.79 ± 30.82)s was significantly shorter than TL group(285.12 ± 52.24)s(P<0.01).The pathological changes of the closed ends were mainly presented as acute thermal and pressure induced injury.Conclusion:Outcomes of the current in vivo experiment suggest that HFEW is an effective and safe method for ligation of small bowel in porcine.Section Ⅳ:Anti-Warburg Effect of Rosmarinic Acid via miR-155 in gastric cancer cellsObjective:The Warburg effect,glycolytic production of ATP under aerobic condition,is a universal property of most cancer cells.Chronic inflammation is a key promoting factor of Warburg effect.Our study was aimed to determine whether rosmarinic acid(RA)had the anti-Warburg effect activity against gastric carcinoma in vitro and in vivo.Meanwhile,the mechanism for anti-Warburg effect would be further investigated.Methods:MTT assay was for MKN45 cell growth assay in vitro.Enzyme-linked immunosorbent assay(ELISA)was for pro-inflammatory cytokines.Realtime PCR was used to evaluate levels of microRNA expressions in cells.Expreesions of proteins were determined by Western blotting.Mouse xenograft models were established with MKN45 cells to access anti-Warburg effect activity against gastric carcinoma in vivo.Results:Our study provided evidence that RA played as a candidate suppressor on Warburg effect.It demonstrated the ability to suppress glucose uptake and lactate production,meanwhile,RA inhibited expression of transcription factor HIF-1α that affected glycolytic pathway.Inflammation promoted Warburg effect of cancer cells.As we had expected,RA could inhibit pro-inflammatory cytokines and microRNAs related to inflammation,suggesting that RA may suppress Warburg effect by inflammatory pathway,such as IL-6/STAT3 pathway.MiR-155 was considered as an important mediator in this relationship between inflammation and tumourigenesis.We further showed that miR-155 was the target gene for regulating Warburg effect through the mechanism of inactivating the IL-6/STAT3 pathway.Moreover,we found RA suppressed Warburg effect in vivo.Conclusions:RA might be therapeutic potential agent against Warburg effect of gastric carcinoma. |
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