This study follows the guidelines of the Ethics Committee of Shunde Hospital of Southern Medical University. Informed consent was obtained from the patient before the surgery for the data and video.
1. Patient selection
<ol>
	<li>Apply this surgical method in the following cases:
	<ol>
		<li>Include patients experiencing abdominal pain or discomfort, coupled with radiological examinations confirming the presence of benign lesions.</li>
		<li>Do not impose specific restrictions on tumor size, but maintain a residual spleen volume exceeding 25%.</li>
		<li>Ensure patients exhibit normal levels of AFP, CEA, and CA-199 or confirm the absence of malignancy.</li>
		<li>Consider obesity, which results in increased visceral fat potentially affecting intraoperative anatomy, but does not view it as an absolute limitation.</li>
	</ol>
	</li>
	<li>Exclude patients from this surgery in the following conditions:
	<ol>
		<li>Exclude if the patient is highly suspected of splenic metastasis from malignant tumors.</li>
		<li>Exclude patients with splenomegaly resulting from hematologic diseases or lymphoma.</li>
		<li>Exclude patients with splenomegaly secondary to liver cirrhosis.</li>
		<li>Exclude patients having life-threatening traumatic spleen rupture.</li>
		<li>Exclude patients who have poor overall health and inability to withstand surgery.</li>
	</ol>
	</li>
</ol>
2. Surgical technique
<ol>
	<li>Set up for the operation.
	<ol>
		<li>Position the patient under general anesthesia in a reverse Trendelenburg posture with the left side of the body inclined approximately 10-30&#176;. 
		NOTE: Make adjustments in position as needed throughout the surgical procedure.</li>
		<li>Position the first surgeon and the assistant with the laparoscope on the patient&#39;s right side and the first assistant on the patient&#39;s left side.</li>
		<li>Establish and maintain pneumoperitoneum at 12-14 mmHg.</li>
		<li>Install four ports on the abdominal wall with the assistance of laparoscopic visualization, as depicted in Figure 1.</li>
	</ol>
	</li>
	<li>Examine the peritoneal cavity under laparoscopy to confirm the absence of malignancies.</li>
	<li>Perform temporary occlusion of the splenic artery.
	<ol>
		<li>Use a vessel sealing system to dissect the gastrosplenic ligament along the greater curvature of the stomach,entering the lesser sac.</li>
		<li>Grasp the stomach and move it to the upper right for better exposure to the surgical field.</li>
		<li>Meticulously dissect the main trunk of the splenic artery at the superior edge of the pancreas and temporarily occlude it with a bulldog clip. 
		NOTE: Following the occlusion of blood flow, the spleen exhibits a decrease in volume and a softer texture, which will provide adequate surgical space.</li>
	</ol>
	</li>
	<li>Dissect the perisplenic ligaments, including the splenocolic, splenorenal, and splenophrenic ligaments. 
	NOTE: Caution must be taken when separating the pancreatic tail from the spleen, particularly in cases of splenomegaly or massive splenic lesions, to prevent uncontrolled hemorrhage and the occurrence of postoperative pancreatic fistula (POPF).</li>
	<li>Perform temporary occlusion of the splenic hilum.
	<ol>
		<li>Expose the splenic hilum by resecting the attachments around it with an ultrasonic scalpel, then implement a temporary occlusion of the splenic hilum with a bulldog clip. 
		NOTE: In cases with variations in splenic blood vessels, merely occluding the splenic artery may not achieve the desired splenic ischemic changes.</li>
		<li>Following this step, conduct a thorough reassessment of the spleen&#39;s color, size, and texture.</li>
	</ol>
	</li>
	<li>Perform intraoperative ultrasound for lesion boundary identification.
	<ol>
		<li>Use intraoperative ultrasound to identify the boundary of the lesion during surgery.</li>
		<li>Apply electrocautery to mark the demarcation line at least 1 cm away from the lesion boundary.</li>
	</ol>
	</li>
	<li>Perform spleen parenchyma dissection.
	<ol>
		<li>Insert a bipolar radiofrequency device into the splenic parenchyma along the demarcation line for coagulation and ablation, setting the radiofrequency energy at 80 W.</li>
		<li>Use an ultrasonic scalpel to dissect the splenic parenchyma in the necrotic coagulation zone.</li>
		<li>Securely clamp thick ducts using Hem-o-lok vascular clips and then carefully cut, ensuring a safe resection of the upper spleen containing the lesion. 
		NOTE:Vessels supplying the removed portion of the spleen need to be carefully ligated and transected</li>
	</ol>
	</li>
	<li>Remove the specimen.
	<ol>
		<li>Release the bulldog clip, ensuring no bleeding from the splenic cut edge, and confirm adequate blood supply to the remnant spleen.</li>
		<li>Use bipolar electrocautery to cauterize the splenic cut edge and apply absorbable hemostatic agents over it.</li>
		<li>Position a drain tube in the splenic fossa.</li>
		<li>Place the specimen into a specimen bag, fragment it with oval forceps, and remove it through the enlarged port.</li>
		<li>Release the pneumoperitoneum and suture the puncture wounds.</li>
	</ol>
	</li>
</ol>
3. Postoperative details
<ol>
	<li>Perform continuous electrocardiography for 24 h postoperatively. Monitor essential signs, including heart rate, blood pressure, respiration, oxygen saturation, central venous pressure, pupillary response, and level of consciousness, todetect early postoperative complications in a timely manner.</li>
	<li>Administer antibiotics (cefuroxime sodium, 1.5 g every 12 h) and proton pump inhibitors routinely following the surgery.</li>
	<li>Remove the gastric tube on the second postoperative day and allow the patient a full liquid diet.</li>
	<li>Remove the abdominal drain tube when the drainage output is less than 100 mL/24 h.</li>
	<li>Schedule a follow-up abdominal CT scan 2 months after the surgery to assess the abdominal condition.</li>
</ol>

Optimized Hemorrhage Control Method for Laparoscopic Partial Splenectomy

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	<li>Lewis, S. M., Williams, A., Eisenbarth, S. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Structure+and+function+of+the+immune+system+in+the+spleen.">Structure and function of the immune system in the spleen.</a> Sci Immunol. 4 (33), eaau6085 (2019).</li><li>Kristinsson, S. Y., Gridley, G., Hoover, R. 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J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+spleen+in+local+and+systemic+regulation+of+immunity.">The spleen in local and systemic regulation of immunity.</a> Immunity. 39 (5), 806-818 (2013).</li><li>Christo, M. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Partial+regulated+splenectomies.+Preliminary+note+on+the+first+3+cases+operated+on.">Partial regulated splenectomies. Preliminary note on the first 3 cases operated on.</a> Hospital (Rio J). 56, 645-650 (1959).</li><li>Redmond, H. P., Redmond, J. M., Rooney, B. P., Duignan, J. P., Bouchier-Hayes, D. 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A review of the first 457 published cases.</a> J Laparoendosc Adv Surg Tech A. 31 (10), 1130-1142 (2021).</li><li>Buzel&#233;, R., Barbier, L., Sauvanet, A., Fantin, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Medical+complications+following+splenectomy.">Medical complications following splenectomy.</a> J Visc Surg. 153 (4), 277-286 (2016).</li><li>Bisharat, N., Omari, H., Lavi, I., Raz, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Risk+of+infection+and+death+among+post-splenectomy+patients.">Risk of infection and death among post-splenectomy patients.</a> J Infect. 43 (3), 182-186 (2001).</li><li>Kruetzmann, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Human+immunoglobulin+M+memory+B+cells+controlling+Streptococcus+pneumoniae+infections+are+generated+in+the+spleen.">Human immunoglobulin M memory B cells controlling Streptococcus pneumoniae infections are generated in the spleen.</a> J Exp Med. 197 (7), 939-945 (2003).</li><li>Leone, G., Pizzigallo, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bacterial+infections+following+splenectomy+for+malignant+and+nonmalignant+hematologic+diseases.">Bacterial infections following splenectomy for malignant and nonmalignant hematologic diseases.</a> Mediterr J Hematol Infect Dis. 7 (1), e2015057 (2015).</li><li>Romano, F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Thrombosis+of+the+splenoportal+axis+after+splenectomy.">Thrombosis of the splenoportal axis after splenectomy.</a> Langenbecks Arch Surg. 391 (5), 483-488 (2006).</li><li>Watters, J. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Splenectomy+leads+to+a+persistent+hypercoagulable+state+after+trauma.">Splenectomy leads to a persistent hypercoagulable state after trauma.</a> Am J Surg. 199 (5), 646-651 (2010).</li><li>Poulin, E. C., Thibault, C., DesC&#244;teaux, J. G., C&#244;t&#233;, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Partial+laparoscopic+splenectomy+for+trauma:+technique+and+case+report.">Partial laparoscopic splenectomy for trauma: technique and case report.</a> Surg Laparosc Endosc. 5 (4), 306-310 (1995).</li><li>Ignjatovic, D., Stimec, B., Zivanovic, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+basis+for+splenic+segmental+dearterialization:+a+post-mortem+study.">The basis for splenic segmental dearterialization: a post-mortem study.</a> Surg Radiol Anat. 27 (1), 15-18 (2005).</li><li>Costi, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Partial+splenectomy:+Who,+when+and+how.+A+systematic+review+of+the+2130+published+cases.">Partial splenectomy: Who, when and how. A systematic review of the 2130 published cases.</a> J Pediatr Surg. 54 (8), 1527-1538 (2019).</li><li>Borie, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Laparoscopic+partial+splenectomy:+Surgical+technique.">Laparoscopic partial splenectomy: Surgical technique.</a> J Visc Surg. 153 (5), 371-376 (2016).</li><li>Ouyang, G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Laparoscopic+partial+splenectomy+with+temporary+occlusion+of+the+trunk+of+the+splenic+artery+in+fifty-one+cases:+experience+at+a+single+center.">Laparoscopic partial splenectomy with temporary occlusion of the trunk of the splenic artery in fifty-one cases: experience at a single center.</a> Surg Endosc. 35 (1), 367-373 (2021).</li><li>Mignon, F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preoperative+selective+embolization+allowing+a+partial+splenectomy+for+splenic+hamartoma.">Preoperative selective embolization allowing a partial splenectomy for splenic hamartoma.</a> Ann Chir. 128 (2), 112-116 (2003).</li><li>Zheng, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Treatment+of+hemangioma+of+the+spleen+by+preoperative+partial+splenic+embolization+plus+laparoscopic+partial+splenectomy:+A+case+report.">Treatment of hemangioma of the spleen by preoperative partial splenic embolization plus laparoscopic partial splenectomy: A case report.</a> Medicine (Baltimore). 97 (17), e0498 (2018).</li><li>Vasilescu, C., Stanciulea, O., Tudor, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Laparoscopic+versus+robotic+subtotal+splenectomy+in+hereditary+spherocytosis.+Potential+advantages+and+limits+of+an+expensive+approach.">Laparoscopic versus robotic subtotal splenectomy in hereditary spherocytosis. Potential advantages and limits of an expensive approach.</a> Surg Endosc. 26 (10), 2802-2809 (2012).</li><li>Wang, W. -. D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Partial+splenectomy+using+a+laparoscopic+bipolar+radiofrequency+device:+a+case+report.">Partial splenectomy using a laparoscopic bipolar radiofrequency device: a case report.</a> World J Gastroenterol. 21 (11), 3420-3424 (2015).</li><li>Liu, D. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Anatomy+of+vasculature+of+850+spleen+specimens+and+its+application+in+partial+splenectomy.">Anatomy of vasculature of 850 spleen specimens and its application in partial splenectomy.</a> Surgery. 119 (1), 27-33 (1996).</li><li>Habib, N. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=How+we+do+a+bloodless+partial+splenectomy.">How we do a bloodless partial splenectomy.</a> Am J Surg. 186 (2), 164-166 (2003).</li><li>Gumbs, A. A., Bouhanna, P., Bar-Zakai, B., Briennon, X., Gayet, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Laparoscopic+partial+splenectomy+using+radiofrequency+ablation.">Laparoscopic partial splenectomy using radiofrequency ablation.</a> J Laparoendosc Adv Surg Tech A. 18 (4), 611-613 (2008).</li><li>Liu, G., Fan, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Feasibility+and+safety+of+laparoscopic+partial+splenectomy:+A+systematic+review.">Feasibility and safety of laparoscopic partial splenectomy: A systematic review.</a> World J Surg. 43 (6), 1505-1518 (2019).</li><li>Teperman, S. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bloodless+splenic+surgery:+The+safe+warm-ischemic+time.">Bloodless splenic surgery: The safe warm-ischemic time.</a> J Pediatr Surg. 29 (1), 88-92 (1994).</li></ol>

For years, total splenectomy was the primary treatment for splenic tumors, splenomegaly, and hematological disorders. However, with extensive cases followed up, complications after total splenectomy, including infectious complications and thromboembolic complications, have gradually aroused attention8. Overwhelming post-splenectomy infections (OPSIs) are the most severe complication after splenectomy, characterized by rapid disease progression with a mortality rate of approximately 50%<sup class='...

With a profound understanding of the physiological functions of the spleen, the research underscores its pivotal role in the body's immune response, hematopoiesis, and clearance of red blood cells1. Complications following splenectomy, such as overwhelming post-splenectomy infections (OPSIs), pulmonary hypertension, and thromboembolism, significantly influence the choice of surgical methods in clinical practice2,3. According to the literature, patients after total splenectomy exhibit a decreased capacity to clear malaria-parasitized RBCs and a higher risk of developing meningitis and sepsis following infections with Streptococcus pneumoniae, Neisseria meningitidis, and Haemophilus influenzae type B4. PS preserves splenic function while ensuring treatment effectiveness, making it widely applied in clinical practice.
In 1959, the first successful PS was reported by Cristo5. The spleen is a fragile organ comprising well-defined splenic segments, each with its distinctive arterial and venous supply, demarcated by relatively avascular regions6. These factors collectively establish the anatomical foundation for PS. However, conventional open surgery carries inherent drawbacks, including significant trauma, cosmetic disadvantages, and postoperative pain. In recent years, alongside the maturation of laparoscopic instruments and techniques, LPS has emerged as the preferred therapeutic modality for benign splenic lesions. Nonetheless, due to the spleen's rich blood supply, substantial intraoperative hemorrhage during laparoscopy may cause a conversion to open surgery. Romboli et al. reviewed 457 cases of LPS, revealing an average operative time of approximately 128 &plusmn; 43.7 min, and demonstrated that about 3.9% of patients required conversion due to hemorrhage, and the average postoperative stay is 4.9 &plusmn; 3.8 days7. Comprehensive knowledge of splenic anatomy and meticulous surgical skills have hindered the broad clinical application of LPS.
To mitigate the risk of intraoperative hemorrhage and expedite the learning curve, we try to perform LPS with complete blood flow occlusion. In this study, we present a 72-year-old female patient with a massive splenic vascular tumor located in the upper middle pole of the spleen and adjacent to the splenic hilum. This novel technique excels in effective hemorrhage control and ensures safety, efficacy, and a high level of reproducibility.

<table><tbody><tr><td>Absorbable hemostat</td><td>Ethicon, LLC</td><td>W1913T</td><td></td></tr><tr><td>Disposable trocar</td><td>Kangji Medical</td><td>101Y.307,101Y.311</td><td></td></tr><tr><td>Endo bag</td><td>Medtronic</td><td>https://www.medtronic.com/covidien/en-us/search.html#q=endo%20bag</td><td>specimen bag</td></tr><tr><td>Jaw sealer/divider</td><td>Covidien Medical</td><td>LF1737</td><td></td></tr><tr><td>Laparoscopic radiofrequency device</td><td>AngioDynamics, Inc</td><td>Rita 700-103659</td><td></td></tr><tr><td>Laparoscopic system</td><td>Olympus</td><td>WM-NP2 L-RECORDOR-01</td><td></td></tr><tr><td>LigaSure&amp;nbsp;</td><td>Medtronic</td><td>https://www.medtronic.com/covidien/en-us/products/vessel-sealing/ligasure-technology.html</td><td>vessel sealing system</td></tr><tr><td>Ligation clips (Hem-o-lok)</td><td>Teleflex Medical</td><td>544240,544230,544220</td><td></td></tr><tr><td>Ultrasonic scalpel</td><td>ETHICON Medical</td><td>HAR36</td><td></td></tr></tbody></table>

application of laparoscopic partial splenectomy with total blood flow occlusion in benign splenic lesions

Laparoscopic partial splenectomy (LPS) is gradually becoming the preferred method for treating benign splenic lesions. However, due to the abundant blood supply and its soft, fragile tissue texture, especially when the lesion is located near the splenic hilum or is particularly large, performing partial splenectomy (PS) in clinical practice is extremely challenging. Therefore, we have been continuously exploring and optimizing hemorrhage control methods during PS, and we here propose a method to perform LPS with complete spleen blood flow occlusion.
This study describes an optimized approach to control intraoperative hemorrhage during LPS. First, it involves the thorough dissection of the splenic ligaments and careful separation of the pancreatic tail from the spleen. With complete exposure to the splenic hilum, we temporarily occlude the entire blood supply of the spleen using a laparoscopic bulldog clip. Subsequently, we employ intraoperative ultrasound to identify the boundary of the lesion and resect the corresponding portion of the spleen under complete blood flow control. This approach embodies the essence of 'spleen preservation' through effective hemorrhage control and precise resection. It is particularly suitable for laparoscopic surgery and deserves further clinical promotion.

In this case, a 72-year-old female patient was admitted for a massive splenic lesion found on a routine examination at a local hospital. She had a history of previous abdominal surgery. Her medical history was unremarkable, and her BMI was normal (20.1 kg/m2). Abdominal contrast-enhanced CT showed a massive lesion located in the upper middle pole of the spleen, with a diameter of approximately 15 cm (Figure 2). Preoperative assessments revealed no evidence of malignancy. Due to th...

Application of Laparoscopic Partial Splenectomy with Total Blood Flow Occlusion in Benign Splenic Lesions

In this study, we describe an intraoperative hemorrhage control technique for laparoscopic partial splenectomy, improving spleen resection's safety and precision.

Laparoscopic partial splenectomy (LPS) is gradually becoming the preferred method for treating benign splenic lesions. However, due to the abundant blood ...

application-laparoscopic-partial-splenectomy-with-total-blood-flow

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140 Views.  First People's Hospital of Shunde. In this study, we describe an intraoperative hemorrhage control technique for laparoscopic partial splenectomy, improving spleen resection's safety and precision.

Video: Application of Laparoscopic Partial Splenectomy with Total Blood Flow Occlusion in Benign Splenic Lesions

Visualization and Analysis of Blood Flow and Oxygen Consumption in Hepatic Microcirculation: Application to an Acute Hepatitis Model

There is a considerable discrepancy between oxygen supply and demand in the liver because hepatic oxygen consumption is relatively high but about 70% of the hepatic blood supply is poorly oxygenated portal vein blood derived from the gastrointestinal tract and spleen. Oxygen is delivered to hepatocytes by blood flowing from a terminal branch of the portal vein to a central venule via sinusoids, and this makes an oxygen gradient in hepatic lobules. The oxygen gradient is an important physical parameter that involves the expression of enzymes upstream and downstream in hepatic microcirculation, but the lack of techniques for measuring oxygen consumption in the hepatic microcirculation has delayed the elucidation of mechanisms relating to oxygen metabolism in liver. We therefore used FITC-labeled erythrocytes to visualize the hepatic microcirculation and used laser-assisted phosphorimetry to measure the partial pressure of oxygen in the microvessels there. Noncontact and continuous optical measurement can quantify blood flow velocities, vessel diameters, and oxygen gradients related to oxygen consumption in the liver. In an acute hepatitis model we made by administering acetaminophen to mice we observed increased oxygen pressure in both portal and central venules but a decreased oxygen gradient in the sinusoids, indicating that hepatocyte necrosis in the pericentral zone could shift the oxygen pressure up and affect enzyme expression in the periportal zone. In conclusion, our optical methods for measuring hepatic hemodynamics and oxygen consumption can reveal mechanisms related to hepatic disease.

An optical system was developed to visualize hepatic microcirculation with FITC-labeled erythrocytes and to measure the partial pressure of oxygen in the microvessels with laser-assisted phosphorimetry. This method can be used to investigate physiological and pathological mechanisms by analyzing microvascular structure, diameter, blood flow velocity, and oxygen tension.

There is a considerable discrepancy between oxygen supply and demand in the liver because hepatic oxygen consumption is relatively high but about 70% of ...

Embolic Middle Cerebral Artery Occlusion (MCAO) for Ischemic Stroke with Homologous Blood Clots in Rats

Clinically, thrombolytic therapy with use of recombinant tissue plasminogen activator (tPA) remains the most effective treatment for acute ischemic stroke. However, the use of tPA is limited by its narrow therapeutic window and by increased risk of hemorrhagic transformation. There is an urgent need to develop suitable stroke models to study new thrombolytic agents and strategies for treatment of ischemic stroke. At present, two major types of ischemic stroke models have been developed in rats and mice: intraluminal suture MCAO and embolic MCAO. Although MCAO models via the intraluminal suture technique have been widely used in mechanism-driven stroke research, these suture models do not mimic the clinical situation and are not suitable for thrombolytic studies. Among these models, the embolic MCAO model closely mimics human ischemic stroke and is suitable for preclinical investigation of thrombolytic therapy. This embolic model was first developed in rats by Overgaard et al.1 in 1992 and further characterized by Zhang et al. in 19972. Although embolic MCAO has gained increasing attention, there are technical problems faced by many laboratories. To meet increasing needs for thrombolytic research, we present a highly reproducible model of embolic MCAO in the rat, which can develop a predictable infarct volume within the MCA territory. In brief, a modified PE-50 tube is gently advanced from the external carotid artery (ECA) into the lumen of the internal carotid artery (ICA) until the tip of the catheter reaches the origin of the MCA. Through the catheter, a single homologous blood clot is placed at the origin of the MCA. To identify the success of MCA occlusion, regional cerebral blood flow was monitored, neurological deficits and infarct volumes were measured. The techniques presented in this paper should help investigators to overcome technical problems for establishing this model for stroke research.

Embolic Middle Cerebral Artery Occlusion MCAO for Ischemic Stroke with Homologous Blood Clots in Rats

In this paper, we demonstrate a standard method for producing an embolic middle cerebral artery occlusion with homologous blood clots (fibrin-rich) in adult rat. This model closely mimics human ischemic stroke and is suitable for preclinical study of thrombolytic therapy for ischemic stroke.

Clinically, thrombolytic therapy with use of recombinant tissue plasminogen activator (tPA) remains the most effective treatment for acute ischemic ...

Complete and Partial Aortic Occlusion for the Treatment of Hemorrhagic Shock in Swine

Hemorrhage remains the leading cause of preventable deaths in trauma. Endovascular management of non-compressible torso hemorrhage has been at the forefront of trauma care in recent years. Since complete aortic occlusion presents serious concerns, the concept of partial aortic occlusion has gained a growing attention. Here, we present a large animal model of hemorrhagic shock to investigate the effects of a novel partial aortic balloon occlusion catheter and compare it with a catheter that works on the principles of complete aortic occlusion. Swine are anesthetized and instrumented in order to conduct controlled fixed-volume hemorrhage, and hemodynamic and physiological parameters are monitored. Following hemorrhage, aortic balloon occlusion catheters are inserted and inflated in the supraceliac aorta for 60 min, during which the animals receive whole-blood resuscitation as 20% of the total blood volume (TBV). Following balloon deflation, the animals are monitored in a critical care setting for 4 h, during which they receive fluid resuscitation and vasopressors as needed. The partial aortic balloon occlusion demonstrated improved distal mean arterial pressures (MAPs) during the balloon inflation, decreased markers of ischemia, and decreased fluid resuscitation and vasopressor use. As swine physiology and homeostatic responses following hemorrhage have been well-documented and are like those in humans, a swine hemorrhagic shock model can be used to test various treatment strategies. In addition to treating hemorrhage, aortic balloon occlusion catheters have become popular for their role in cardiac arrest, cardiac and vascular surgery, and other high-risk elective surgical procedures.

Here, we present a protocol demonstrating a hemorrhagic shock model in swine that uses aortic occlusion as a bridge to definitive care in trauma. This model has application in testing a wide range of surgical and pharmacological therapeutic strategies.

Hemorrhage remains the leading cause of preventable deaths in trauma. Endovascular management of non-compressible torso hemorrhage has been at the ...

Quantitative Analysis of Cellular Composition in Advanced Atherosclerotic Lesions of Smooth Muscle Cell Lineage-Tracing Mice

Atherosclerosis remains the leading cause of death worldwide and, despite countless preclinical studies describing promising therapeutic targets, novel interventions have remained elusive. This is likely due, in part, to a reliance on preclinical prevention models investigating the effects of genetic manipulations or pharmacological treatments on atherosclerosis development rather than the established disease. Also, results of these studies are often confounding because of the use of superficial lesion analyses and a lack of characterization of lesion cell populations. To help overcome these translational hurdles, we propose an increased reliance on intervention models that employ investigation of changes in cellular composition at a single cell level by immunofluorescent staining and confocal microscopy. To this end, we describe a protocol for testing a putative therapeutic agent in a murine intervention model including a systematic approach for animal dissection, embedding, sectioning, staining, and quantification of brachiocephalic artery lesions. In addition, due to the phenotypic diversity of cells within late-stage atherosclerotic lesions, we describe the importance of using cell-specific, inducible lineage tracing mouse systems and how this can be leveraged for unbiased characterization of atherosclerotic lesion cell populations. Together, these strategies may assist vascular biologists to more accurately model therapeutic interventions and analyze atherosclerotic disease and will hopefully translate into a higher rate of success in clinical trials.

We propose a standardized protocol to characterize the cellular composition of late-stage murine atherosclerotic lesions including systematic methods of animal dissection, tissue embedding, sectioning, staining, and analysis of brachiocephalic arteries from atheroprone smooth muscle cell lineage tracing mice.

Atherosclerosis remains the leading cause of death worldwide and, despite countless preclinical studies describing promising therapeutic targets, novel ...

A Rat Carotid Artery Pressure-Controlled Segmental Balloon Injury with Periadventitial Therapeutic Application

Cardiovascular disease remains the leading cause of death and disability worldwide, in part due to atherosclerosis. Atherosclerotic plaque narrows the luminal surface area in arteries thereby reducing adequate blood flow to organs and distal tissues. Clinically, revascularization procedures such as balloon angioplasty with or without stent placement aim to restore blood flow. Although these procedures reestablish blood flow by reducing plaque burden, they damage the vessel wall, which initiates the arterial healing response. The prolonged healing response causes arterial restenosis, or re-narrowing, ultimately limiting the long-term success of these revascularization procedures. Therefore, preclinical animal models are integral for analyzing the pathophysiological mechanisms driving restenosis, and provide the opportunity to test novel therapeutic strategies. Murine models are cheaper and easier to operate on than large animal models. Balloon or wire injury are the two commonly accepted injury modalities used in murine models. Balloon injury models in particular mimic the clinical angioplasty procedure and cause adequate damage to the artery for the development of restenosis. Herein we describe the surgical details for performing and histologically analyzing the modified, pressure-controlled rat carotid artery balloon injury model. Additionally, this protocol highlights how local periadventitial application of therapeutics can be used to inhibit neointimal hyperplasia. Lastly, we present light sheet fluorescence microscopy as a novel approach for imaging and visualizing the arterial injury in three-dimensions.

The rat carotid artery balloon injury mimics the clinical angioplasty procedure performed to restore blood flow in atherosclerotic vessels. This model induces the arterial injury response by distending the arterial wall, and denuding the intimal layer of endothelial cells, ultimately causing remodeling and an intimal hyperplastic response.

Cardiovascular disease remains the leading cause of death and disability worldwide, in part due to atherosclerosis. Atherosclerotic plaque narrows the ...

Hemocompatibility Testing of Blood-Contacting Implants in a Flow Loop Model Mimicking Human Blood Flow

The growing use of medical devices (e.g., vascular grafts, stents, and cardiac catheters) for temporary or permanent purposes that remain in the body&#39;s circulatory system demands a reliable and multiparametric approach that evaluates the possible hematologic complications caused by these devices (i.e., activation and destruction of blood components). Comprehensive in vitro hemocompatibility testing of blood-contacting implants is the first step towards successful in vivo implementation. Therefore, extensive analysis according to the International Organization for Standardization 10993-4 (ISO 10993-4) is mandatory prior to clinical application. The presented flow loop describes a sensitive model to analyze the hemostatic performance of stents (in this case, neurovascular) and reveal adverse effects. The use of fresh human whole blood and gentle blood sampling are essential to avoid the preactivation of blood. The blood is perfused through a heparinized tubing containing the test specimen by using a peristaltic pump at a rate of 150 mL/min at 37 &#176;C for 60 min. Before and after perfusion, hematologic markers (i.e., blood cell count, hemoglobin, hematocrit, and plasmatic markers) indicating the activation of leukocytes (polymorphonuclear [PMN]-elastase), platelets (&#946;-thromboglobulin [&#946;-TG]), the coagulation system (thombin-antithrombin III [TAT]), and the complement cascade (SC5b-9) are analyzed. In conclusion, we present an essential and reliable model for extensive hemocompatibility testing of stents and other blood-contacting devices prior to clinical application.

This protocol describes a comprehensive hemocompatibility evaluation of blood-contacting devices using laser-cut neurovascular implants. A flow loop model with fresh, heparinized human blood is applied to mimic blood flow. After perfusion, various hematologic markers are analyzed and compared to the values gained directly after blood collection for hemocompatibility evaluation of the tested devices.

The growing use of medical devices (e.g., vascular grafts, stents, and cardiac catheters) for temporary or permanent purposes that remain in the ...

Bloodless Laparoscopic Partial Splenectomy Assisted by Bipolar Radiofrequency Excision Hemostatic Device

Among the lymphatic system in the human body, the spleen is the most extensive one and has hematopoietic, hemofiltration, blood storage, and immune functions. As a new method of preserving the spleen, laparoscopic partial splenectomy (LPS) has been increasingly applied in clinical practice with people&#39;s deeper insights into minimally invasive treatment and the development of technical equipment. Compared with conventional open splenectomy, LPS can preserve normal spleen tissue as much as possible, decrease the occurrence of complications after total splenectomy, and reduce postoperative hospital stay. The bipolar radiofrequency excision hemostatic device used for LPS can solidify the splenic tissue and close the small blood vessels, which reduces the hemorrhage of the spleen cross-section and clears the operative field, thus achieving the ideal effect of &#34;bloodless partial splenectomy&#34;. Therefore, under the premise of strictly mastering the indications and fully understanding the vascular anatomy of the spleen, the application of the bipolar radiofrequency excision hemostatic&#160;device in LPS is worthy of clinical promotion.

The present protocol describes that bipolar radiofrequency device-assisted laparoscopic partial splenectomy (LPS) is safe and effective. A bipolar radiofrequency device in LPS can reduce intraoperative bleeding and achieve the clinical effect of a "bloodless spleen incision", which is worthy of clinical application.

Among the lymphatic system in the human body, the spleen is the most extensive one and has hematopoietic, hemofiltration, blood storage, and immune ...

Modified Laparoscopic Anatomic Hepatectomy: Two-Surgeon Technique Combined with the Simple Extracorporeal Pringle Maneuver

Laparoscopic anatomic hepatectomy (LAH) has become increasingly prevalent worldwide in recent years. However, LAH remains a challenging procedure due to the anatomical characteristics of the liver, with intraoperative hemorrhage being a primary concern. Intraoperative blood loss is the leading cause of conversion to open surgery; therefore, effective management of bleeding and hemostasis is crucial for a successful LAH.
The two-surgeon technique is proposed as an alternative to the traditional single-surgeon approach, with potential benefits in reducing intraoperative bleeding during laparoscopic hepatectomy. However, there remains a lack of evidence to determine which mode of the two-surgeon technique yields superior patient outcomes. Besides, to our knowledge, the LAH technique, which involves the use of a cavitron ultrasonic surgical aspirator (CUSA) by the primary surgeon while an ultrasonic dissector by the second surgeon, has been rarely reported before.
Herein, we present a modified, two-surgeon LAH technique, wherein one surgeon employs a CUSA while the other uses an ultrasonic dissector. This technique is combined with a simple extracorporeal Pringle maneuver and low central venous pressure (CVP) approach. In this modified technique, the primary and secondary surgeons utilize a laparoscopic CUSA and an ultrasonic dissectorconcurrently to achieve precise and expeditious hepatectomy. A simple extracorporeal Pringle maneuver, combined with the maintenance of low CVP, is employed to regulate the hepatic inflow and outflow in order to minimize intraoperative bleeding. This approach facilitates the attainment of a dry and clean operative field, which allows for the precise ligation and dissection of blood vessels and bile ducts. The modified LAH procedure is simpler and safer due to its effective control over bleeding as well as the seamless transition between the roles of primary and secondary surgeons. It holds great promise for future clinical applications.

Here, we present a protocol to perform a modified laparoscopic anatomic hepatectomy using improved techniques and instruments.

Laparoscopic anatomic hepatectomy (LAH) has become increasingly prevalent worldwide in recent years. However, LAH remains a challenging procedure due to ...

Reducing Bleeding in Laparoscopic Partial Splenectomy with Microwave Ablation

Application of Microwave Ablation in Laparoscopic Partial Splenectomy

Splenic hemangioma is the most common pathological classification of splenic tumors, and its surgical indication and treatment have been controversial. Before, open splenectomy was usually used to treat splenic hemangioma. Following the rapid development of laparoscopic techniques, people's requirements for minimally invasive treatment have gradually increased, and laparoscopic splenectomy has gradually become the main treatment method. However, through the deeper study of spleen function, it was found that partial splenectomy can cut down the incidence of postoperative thrombocythemia and decrease side effects on the physiological function of the body, so laparoscopic partial splenectomy came into being. However, due to the special anatomical structure, the incidence of hemorrhage during partial splenectomy is greater. Therefore, during the operation, we removed part of the splenic blood vessels, combined with microwave ablation, which perfectly solved the problem of intraoperative bleeding. Laparoscopic partial splenectomy combined with microwave ablation not only achieves the requirements of minimally invasive treatment but also reduces the risk of intraoperative bleeding, meriting clinical application and promotion.

The use of laparoscopic partial splenectomy has been limited due to the high risk of bleeding during surgery. Therefore, we introduce a laparoscopic method combined with microwave ablation to solve the problem of intraoperative bleeding.

Splenic hemangioma is the most common pathological classification of splenic tumors, and its surgical indication and treatment have been controversial. ...

Laparoscopic Splenectomy with Pericardial Devascularization for Hypersplenism and Esophageal Variceal Hemorrhage Due to Portal Hypertension

Hypersplenism and esophageal variceal hemorrhage caused by portal hypertension are common and serious complications of decompensated cirrhosis. In recent years, with the widespread application of various therapeutic methods such as drugs, endoscopy, splenic artery embolization, transjugular intrahepatic portal shunt, and liver transplantation, the role of surgery in the treatment of portal hypertension has gradually diminished, and the indications for surgical treatment have become more strictly defined. However, according to the clinical practice in China, surgical treatment of portal hypertension still holds an important role that other treatments cannot fully replace. In fact, surgical treatment of portal hypertension is widely performed in hospitals at all levels in China, saving numerous lives. Splenectomy combined with pericardial devascularization (SPD) is the most common surgical method for treating hypersplenism and esophageal variceal hemorrhage caused by portal hypertension. Long-term clinical practice has proven that SPD is a safe and effective treatment for hypersplenism and esophageal variceal rupture and hemorrhage due to portal hypertension. With the rapid development of laparoscopic techniques, the minimally invasive advantages of laparoscopic splenectomy combined with pericardial devascularization (LSPD) have become increasingly evident. However, the successful performance of LSPD mainly depends on the skill and proficiency of the surgeon. In this context, this article presents detailed techniques for LSPD.

With the rapid advancement of laparoscopic techniques, the minimally invasive benefits of laparoscopic splenectomy combined with pericardial devascularization have become increasingly evident. In this context, we present a protocol for performing laparoscopic splenectomy alongside pericardial devascularization to treat hypersplenism and esophageal variceal hemorrhage resulting from portal hypertension.

Hypersplenism and esophageal variceal hemorrhage caused by portal hypertension are common and serious complications of decompensated cirrhosis. In recent ...