We thank ZHZ for the conception and design of the study. JW and YPZ performed and supervised the operation. YH, YBX, SL contributed to data analysis. JW and WJL prepared the draft of the video. All authors participated in the revision of the manuscript. All authors read and approved the final manuscript.

All methods described here were approved by the Ethics Committee of Union Hospital Tongji Medical College, Huazhong University of Science and Technology. We obtained the patient&#39;s consent to use his surgical footage as a video after the procedure.
1. Instruments for operation
<ol>
	<li>Ensure the availability of a surgical microscope system, microsurgical instrument, and microscopic bipolar electrocoagulation.</li>
</ol>
2. Preparation for operation
<ol>
	<li>Inclusion criteria
	<ol>
		<li>Include all patients diagnosed with azoospermia who agree to undergo surgery. Have all female partners of the patients undergo gynecologic fertility testing 
		NOTE: In the presented study, patients underwent surgery at the hospital between June 2013 and July 2020. All female patients&#39; partners&#39; gynecologic fertility test results were normal.</li>
		<li>Ensure normal serum levels of follicle-stimulating hormone and testosterone in all patients.</li>
		<li>Ensure that there are no genital malformations, hypertension, diabetes, hyperthyroidism, heart disease, pelvic trauma, abnormal psychological or mental disorders, or other serious preexisting diseases or conditions.</li>
	</ol>
	</li>
	<li>Exclusion criteria
	<ol>
		<li>Exclude patients with genital malformations, abnormal psychological or mental disorders, acute urinary tract infection or urinary tuberculosis, severe abdominal trauma or surgery, severe coagulation disorders, or other contraindications to anesthesia or laparoscopy.</li>
	</ol>
	</li>
	<li>Perform an anesthesia risk assessment on patients prior to the surgery. Ask patients who have been taking aspirin to discontinue it.</li>
	<li>Mark the skin 15 cm around the perineum with a skin preparation knife to prepare for the surgery.</li>
	<li>Administer antibiotics to the patient via intravenous drip 30 min prior to surgery. Ideally, use cefoperazone sodium (2.0 g) together with 100 mL of 0.9% sodium chloride solution.</li>
	<li>Make sure that the patient is under general or intraspinal anesthesia.</li>
</ol>
3. Procedure
<ol>
	<li>Place the patient under general anesthesia with endotracheal intubation. Then, perform standard disinfection with 1% iodophor while the patient is in the supine position.</li>
	<li>Make a 3 cm incision along the mid-scrotal fold. Then, pinch one testicle and incise the flesh membrane to expose the testicular sheath and spermatic cord.</li>
	<li>Dissect the fascia between the vas deferens and the spermatic cord vessels. Separate along the space between the vas deferens vasculature and the vasculature of the spermatic cord with a vascular clamp. Secure the vas deferens with tissue forceps and prepare it for vesicocentesis.</li>
	<li>Insert a 24 G cannula needle obliquely 15&#176; downward along the long axis of the vas deferens. Pull out the needle core after the obvious tactile breakthrough, and slide the soft trocar to the lumen.</li>
	<li>Inject 2 mL of 0.9% sodium chloride solution into the vas deferens using a 5 mL syringe connected to the 24 G cannula. If resistance is high, this confirms that there is an obstruction in the vas deferens.</li>
	<li>Take a sample of fluid from the syringe and give it to an examiner to examine it under the microscope for sperm.</li>
	<li>Advance the 3-0 polypropylene suture through the trocar. If there is resistance, it indicates obstruction. Measure the length of the suture to determine the site of obstruction.</li>
</ol>
4. Postoperative care
<ol>
	<li>Make the patient stay in the ICU for ~1-2 h postoperatively, until he fully regains consciousness.</li>
	<li>Send the patient back to the ward when he fully wakes up.</li>
	<li>Implant a catheter for 6-24 h.</li>
	<li>Lift the scrotum with underwear for 1 week.</li>
</ol>

<ol>
	<li>Chen, X. 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=Clinical+features+and+therapeutic+strategies+of+obstructive+azoospermia+in+patients+treated+by+bilateral+inguinal+hernia+repair+in+childhood.">Clinical features and therapeutic strategies of obstructive azoospermia in patients treated by bilateral inguinal hernia repair in childhood.</a> Asian Journal of Andrology. 16 (5), 745-748 (2014).</li><li>Sparkman, R. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bilateral+exploration+in+inguinal+hernia+in+juvenile+patients.+Review+and+appraisal.">Bilateral exploration in inguinal hernia in juvenile patients. Review and appraisal.</a> Surgery. 51, 393-406 (1962).</li><li>Pollak, R., Nyhus, L. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Complications+of+groin+hernia+repair.">Complications of groin hernia repair.</a> The Surgical Clinics of North America. 63 (6), 1363-1371 (1983).</li><li>Sheynkin, Y. R., Hendin, B. N., Schlegel, P. N., Goldstein, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microsurgical+repair+of+iatrogenic+injury+to+the+vas+deferens.">Microsurgical repair of iatrogenic injury to the vas deferens.</a> The Journal of Urology. 159 (1), 139-141 (1998).</li><li>Wang, J., 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+for+Vas+Deferens+Obstruction+Following+Childhood+Herniorrhaphy.">Treatment for Vas Deferens Obstruction Following Childhood Herniorrhaphy.</a> Urology. 112 (1), 80-84 (2018).</li><li>Smith, R. P., 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=The+significance+of+sperm+heads+and+tails+within+the+vasal+fluid+during+vasectomy+reversal.">The significance of sperm heads and tails within the vasal fluid during vasectomy reversal.</a> Indian Journal of Urology. 30 (2), 164-168 (2014).</li><li>Jungwirth, A., Diemer, T., Kopa, Z., Krausz, C., Tournaye, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=European+Association+of+Urology+guidelines+on+male+infertility.+2017+edition.">European Association of Urology guidelines on male infertility. 2017 edition.</a> , 23-24 (2017).</li><li>Bromage, S. J., Falconer, D. A., Lieberman, B. A., Sangar, V., Payne, S. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sperm+retrieval+rates+in+subgroups+of+primary+azoospermic+males.">Sperm retrieval rates in subgroups of primary azoospermic males.</a> European Urology. 51 (2), 539-540 (2007).</li><li>Du, J., 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=Differential+diagnosis+of+azoospermia+and+etiologic+classification+of+obstructive+azoospermia:+role+of+scrotal+and+transrectal+US.">Differential diagnosis of azoospermia and etiologic classification of obstructive azoospermia: role of scrotal and transrectal US.</a> Radiology. 256 (2), 493-503 (2010).</li><li>Pasqualotto, F. F., Pasqualotto, E. B., Agarwal, A., Thomas Jr, A. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Results+of+microsurgical+anastomosis+in+men+with+seminal+tract+obstruction+due+to+inguinal+herniorrhaphy.">Results of microsurgical anastomosis in men with seminal tract obstruction due to inguinal herniorrhaphy.</a> Revista do Hospital das Clinicas. 58 (6), 305-309 (2003).</li><li>Shaeer, O. K. Z., Shaeer, K. Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Laparoscopy-assisted+pelvi-scrotal+vasovasostomy.">Laparoscopy-assisted pelvi-scrotal vasovasostomy.</a> Andrologia. 36 (5), 311-314 (2004).</li></ol>

The authors have no conflict of interest and nothing to disclose.

As mentioned before, the location of the vas deferens stump cannot be determined at the inguinal incision, which requires exploration of the original inguinal incision. In this study, we can use this simple operation to determine the site of obstruction and make surgical decisions. If the site of obstruction is in the original incision, MVV can be considered. If the vas deferens injury is not found in the inguinal canal, we can use lapa-VV or SV.
When pasty fluid without sperm was found and th...

Microsurgical vasovasostomy (MVV) is a common treatment for vas deferens obstruction after herniorrhaphy in childhood1,2. The site of obstruction cannot be identified preoperatively, and it is not clear whether sperm are present in the vas deferens or not. This may lead to difficulty in selecting the incision and injury to the vas deferens during semi-open exploration, which decreases the success rate of anastomosis, prolongs the operation time, and increases the risk of abdominal exploration3,4. 
Injury to the vas deferens in the inguinal canal is likely an after-effect of inguinal hernia surgery in childhood, which can usually be caused by ligation, incision, or overstretching1. The rate of azoospermia after herniorrhaphy in childhood is approximately 0.8%-2.0% in pediatric patients. However, in the infertile population, the rate of iatrogenic obstruction is significantly increased, with a history of vas deferens injury in 7.2% of andrological procedures2,3. Surgical exploration of the inguinal region is usually performed to look for vas deferens remnants for anastomosis4. In previous cases, the ruptured end of the vas deferens is usually not found during inguinal exploration, which leads to the termination of the operation and increases surgical trauma to the patient. Therefore, it is very important to use a simple method to determine the location of the obstruction to clarify the following surgical procedures. 
In this study, a 24 G cannula was used to puncture the vas deferens to confirm the presence of sperm in it, and a 3-0 polypropylene suture was passed through the cannula to determine the location of the obstructed site. Based on this simple operation, we can determine the next surgical procedure. After practicing and summarizing the clinical operations, our specific surgical method is presented below.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>0.9% sodium chloride solution</td>
			<td>Guangdong Otsuka Pharmaceutical Co. LTD</td>
			<td>&#160;21M1204&#160;</td>
			<td>Inject into the vas deferens</td>
		</tr>
		<tr>
			<td>1% iodophor</td>
			<td>Guangzhou Qingfeng Disinfection Products Co., LTD&#160;</td>
			<td>Q/QFXD2</td>
			<td>Skin disinfection</td>
		</tr>
		<tr>
			<td>24G cannula needle</td>
			<td>Suzhou Linhua Medical Instrument Co., LTD</td>
			<td>20193141896</td>
			<td>Size:24G, Type:Pen type, with wings, with injection port, Y type</td>
		</tr>
		<tr>
			<td>3-0 monofilament polypropylene</td>
			<td>Covidien Medical Equipment international trading (Shanghai) Co., LTD</td>
			<td>DOL2861y</td>
			<td>Taper Point, Size 3-0, Blue, 36&#34;, Needle V-20, 1/2 Circle</td>
		</tr>
		<tr>
			<td>5 mL syringe</td>
			<td>&#160;Kindly Medical, Shanghai</td>
			<td>&#160;K20210826</td>
			<td>Inject 0.9% sodium chloride solution into the vas deferens</td>
		</tr>
		<tr>
			<td>F16 Catheter</td>
			<td>&#160;Guangzhou Weili Co., Ltd&#160;</td>
			<td>20190612</td>
			<td>Drainage of urine</td>
		</tr>
		<tr>
			<td>micro haemostatic forceps</td>
			<td>Shanghai Medical Instrument (Group) Co., LTD.</td>
			<td>W40350</td>
			<td>Used in surgical procedures</td>
		</tr>
		<tr>
			<td>micro scissors</td>
			<td>Shanghai Medical Instrument (Group) Co., LTD.</td>
			<td>WA1010</td>
			<td>Used in surgical procedures</td>
		</tr>
		<tr>
			<td>micro tweezers</td>
			<td>Shanghai Medical Instrument (Group) Co., LTD.</td>
			<td>WA1020</td>
			<td>Used in surgical procedures</td>
		</tr>
		<tr>
			<td>Olympus microscope</td>
			<td>Olympus Corporation</td>
			<td>Scipu001158</td>
			<td>4&#215;N.A.0.10 W.D.22.0mm&#160;&#160;&#160;&#160; 10&#215;N.A.0.25 W.D.10.5mm&#160;&#160;&#160;&#160;&#160;&#160; 40&#215;N.A.0.65 W.D.0.56mm&#160;&#160;&#160;&#160; 100&#215;N.A.1.25 W.D.0.13mm&#160;</td>
		</tr>
		<tr>
			<td>Operating microscope system</td>
			<td>&#160;Carl Zeiss Co., Ltd&#160;</td>
			<td>OPMI VARIO 700</td>
			<td></td>
		</tr>
	</tbody>
</table>

clinical application of 24 g cannula needle and 3-0 polypropylene suture in vas deferens exploration

The goal of this article is to present the application of a 24 G cannula and 3-0 polypropylene suture as a simple method for the exploration of the vas deferens. During the exploration of the vas deferens, a 24 G cannula needle was used to puncture it. The fluid in the smear confirmed the presence of sperm, to determine whether or not there was concomitant obstruction at the junction of the epididymis and the vas deferens. Then, a 3-0 polypropylene suture (this suture specification has the advantages of a smooth surface, robust quality, and can be passed through a 24 G cannula needle) was passed through the cannula needle to probe the location of the obstructed site. With this technique, exploration of the vas deferens could be more targeted and accurate.

There were 67 patients enrolled in the study. As shown in Table 1, the mean age of the patients was 28.8 &#177; 3.7 years (range: 23-45 years). The time since herniorrhaphy was 24.5 &#177; 3.2 years (range: 21-43 years). The female partners were 25.2 &#177; 3.2 years (range: 23-42 years) years old. A total of 11 (16.4%) patients had pain in the scrotum. Four patients (6.0%) were underweight (BMI &#60; 18.5 kg/m2), 53 (79.1%) had normal weight (BMI 18.5-24.9 kg/m2), eight (11.9%) wer...

Watch this Scientific Journal Video about Clinical Application of 24 G Cannula Needle and 3-0 Polypropylene Suture in Vas Deferens Exploration at JoVE.com

Clinical Application of 24 G Cannula Needle and 3-0 Polypropylene Suture in Vas Deferens Exploration

This protocol presents the clinical application of a 24 G cannula and 3-0 polypropylene suture as a simple and effective method for the exploration of the vas deferens.

The goal of this article is to present the application of a 24 G cannula and 3-0 polypropylene suture as a simple method for the exploration of the vas ...

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2.5K Views.  Huazhong University of Science and Technology. This protocol presents the clinical application of a 24 G cannula and 3-0 polypropylene suture as a simple and effective method for the exploration of the vas deferens.

Video: Clinical Application of 24 G Cannula Needle and 3-0 Polypropylene Suture in Vas Deferens Exploration

The Use of Pharmacological-challenge fMRI in Pre-clinical Research: Application to the 5-HT System

Pharmacological MRI (phMRI) is a new and promising method to study the effects of substances on brain function that can ultimately be used to unravel underlying neurobiological mechanisms behind drug action and neurotransmitter-related disorders, such as depression and ADHD. Like most of the imaging methods (PET, SPECT, CT) it represents a progress in the investigation of brain disorders and the related function of neurotransmitter pathways in a non-invasive way with respect of the overall neuronal connectivity. Moreover it also provides the ideal tool for translation to clinical investigations. MRI, while still behind in molecular imaging strategies compared to PET and SPECT, has the great advantage to have a high spatial resolution and no need for the injection of a contrast-agent or radio-labeled molecules, thereby avoiding the repetitive exposure to ionizing radiations. Functional MRI (fMRI) is extensively used in research and clinical setting, where it is generally combined with a psycho-motor task. phMRI is an adaptation of fMRI enabling the investigation of a specific neurotransmitter system, such as serotonin (5-HT), under physiological or pathological conditions following activation via administration of a specific challenging drug.
The aim of the method described here is to assess brain 5-HT function in free-breathing animals. By challenging the 5-HT system while simultaneously acquiring functional MR images over time, the response of the brain to this challenge can be visualized. Several studies in animals have already demonstrated that drug-induced increases in extracellular levels of e.g. 5-HT (releasing agents, selective re-uptake blockers, etc) evoke region-specific changes in blood oxygenation level dependent (BOLD) MRI signals (signal due to a change of the oxygenated/deoxygenated hemoglobin levels occurring during brain activation through an increase of the blood supply to supply the oxygen and glucose to the demanding neurons) providing an index of neurotransmitter function. It has also been shown that these effects can be reversed by treatments that decrease 5-HT availability16,13,18,7. In adult rats, BOLD signal changes following acute SSRI administration have been described in several 5-HT related brain regions, i.e. cortical areas, hippocampus, hypothalamus and thalamus9,16,15. Stimulation of the 5-HT system and its response to this challenge can be thus used as a measure of its function in both animals and humans2,11.

The goal of this technique is to assess serotonin (5-HT) neurotransmitter function in the live and free-breathing animal with pharmacological magnetic resonance imaging (phMRI) and an intravenous challenge with a selective serotonin reuptake inhibitor (SSRI), fluoxetine.

Pharmacological MRI (phMRI) is a new and promising method to study the effects of substances on brain function that can ultimately be used to unravel ...

In Vivo, Percutaneous, Needle Based, Optical Coherence Tomography of Renal Masses

Optical coherence tomography (OCT) is the optical equivalent of ultrasound imaging, based on the backscattering of near infrared light. OCT provides real time images with a 15 &#181;m axial resolution at an effective tissue penetration of 2-3 mm. Within the OCT images the loss of signal intensity per millimeter of tissue penetration, the attenuation coefficient, is calculated. The attenuation coefficient is a tissue specific property, providing a quantitative parameter for tissue differentiation. 
Until now, renal mass treatment decisions have been made primarily on the basis of MRI and CT imaging characteristics, age and comorbidity. However these parameters and diagnostic methods lack the finesse to truly detect the malignant potential of a renal mass. A successful core biopsy or fine needle aspiration provides objective tumor differentiation with both sensitivity and specificity in the range of 95-100%. However, a non-diagnostic rate of 10-20% overall, and even up to 30% in SRMs, is to be expected, delaying the diagnostic process due to the frequent necessity for additional biopsy procedures. 
We aim to develop OCT into an optical biopsy, providing real-time imaging combined with on-the-spot tumor differentiation. This publication provides a detailed step-by-step approach for percutaneous, needle based, OCT of renal masses.

In Vivo, Percutaneous, Needle Based, Optical Coherence Tomography of Renal Masses

Optical coherence tomography (OCT) is a high resolution imaging technique that allows analysis of tissue specific optical properties providing the means for tissue differentiation. We developed needle based OCT, providing real-time imaging combined with on-the-spot tumor differentiation. This publication describes a method for percutaneous, needle based OCT of renal masses.

Optical coherence tomography (OCT) is the optical equivalent of ultrasound imaging, based on the backscattering of near infrared light. OCT provides real ...

Isolation and Culture of Primary Endothelial Cells from Canine Arteries and Veins

Cardiovascular disease is studied in both human and veterinary medicine. Endothelial cells have been used extensively as an in vitro model to study vasculogenesis, (tumor) angiogenesis, and atherosclerosis. The current standard for in vitro research on human endothelial cells (ECs) is the use of Human Umbilical Vein Endothelial Cells (HUVECs) and Human Umbilical Artery Endothelial Cells (HUAECs). For canine endothelial research, only one cell line (CnAOEC) is available, which is derived from canine aortic endothelium. Although currently not completely understood, there is a difference between ECs originating from either arteries or veins. For a more direct approach to in vitro functionality studies on ECs, we describe a new method for isolating Canine Primary Endothelial Cells (CaPECs) from a variety of vessels. This technique reduces the chance of contamination with fast-growing cells such as fibroblasts and smooth muscle cells, a problem that is common in standard isolation methods such as flushing the vessel with enzymatic solutions or mincing the vessel prior to digestion of the tissue containing all cells. The technique we describe was optimized for the canine model, but can easily be utilized in other species such as human.

Novel isolation methods of primary endothelial cells from blood vessels are needed. This protocol describes a new technique that completely inverts blood vessels of interest, exposing only the endothelial side to enzymatic digestion. The resulting pure endothelial cell culture can be used to study cardiovascular diseases, disease modelling, and angiogenesis.

Cardiovascular disease is studied in both human and veterinary medicine. Endothelial cells have been used extensively as an in vitro model to study ...

Autonomic Function Following Concussion in Youth Athletes: An Exploration of Heart Rate Variability Using 24-hour Recording Methodology

Participation in organized sports makes a significant contribution to youth development, but places youth at a higher risk for sustaining a concussion. To date, return-to-activity decision-making has been anchored in the monitoring of self-reported concussion symptoms and neurocognitive testing. However, multi-modal assessments that corroborate objective physiological measures with traditional subjective symptom reporting are needed and can be valuable. Heart rate variability (HRV) is a non-invasive physiological indicator of the autonomic nervous system, capturing the reciprocal interplay between the sympathetic and parasympathetic nervous systems. There is a dearth of literature exploring the effect of concussion on HRV in youth athletes, and developmental differences preclude the application of adult findings to a pediatric population. Further, the current state of HRV methodology has primarily included short-term (5-15 min) recordings, by using resting state or short-term physical exertion testing to elucidate changes following concussion. The novelty in utilizing a 24 h&#160;recording methodology is that it has the potential to capture natural variation in autonomic function, directly related to the activities a youth athlete performs on a regular basis. Within a prospective, longitudinal research setting, this novel approach to quantifying autonomic function can provide important information regarding the recovery trajectory, alongside traditional self-report symptom measures. Our objectives regarding a 24 h recording methodology were to (1) evaluate the physiological effects of a concussion in youth athletes, and (2) describe the trajectory of physiological change, while considering the resolution of self-reported post-concussion symptoms. To achieve these objectives, non-invasive sensor technology was implemented. The raw beat-to-beat time intervals captured can be transformed to derive time domain and frequency domain measures, which reflect an individual&#39;s ability to adapt and be flexible to their ever-changing environment. By using non-invasive heart rate technology, autonomic function can be quantified outside of a traditional controlled research setting.

We demonstrate a 24 h heart rate recording methodology to evaluate the influence of concussion across the recovery trajectory in youth athletes, within an ecologically valid context.

Participation in organized sports makes a significant contribution to youth development, but places youth at a higher risk for sustaining a concussion. To ...

A Model of Glaucoma Induced by Circumlimbal Suture in Rats and Mice

The circumlimbal suture is a technique for inducing experimental glaucoma in rodents by chronically elevating intraocular pressure (IOP), a well-known risk factor for glaucoma. This protocol demonstrates a step-by-step guide on this technique in Long Evans rats and C57BL/6 mice. Under general anesthesia, a "purse-string" suture is applied on the conjunctiva, around the equator and behind the limbus of the eye. The fellow eye serves as an untreated control. Over the duration of our study, which was a period of 8 weeks for rats and 12 weeks for mice, IOP remained elevated, as measured regularly by rebound tonometry in conscious animals without topical anesthesia. In both species, the sutured eyes showed electroretinogram features consistent with preferential inner retinal dysfunction. Optical coherence tomography showed selective thinning of the retinal nerve fiber layer. Histology of the rat retina in cross-section found reduced cell density in the ganglion cell layer, but no change in other cellular layers. Staining of flat-mounted mouse retinae with a ganglion cell specific marker (RBPMS) confirmed ganglion cell loss. The circumlimbal suture is a simple, minimally invasive and cost-effective way to induce ocular hypertension that leads to ganglion cell injury in both rats and mice.

Chronic ocular hypertension is induced by applying a circumlimbal suture in rats and mice, leading to functional and structural deterioration of the retinal ganglion cells consistent with glaucoma.

The circumlimbal suture is a technique for inducing experimental glaucoma in rodents by chronically elevating intraocular pressure (IOP), a well-known ...

Proton Therapy Delivery and Its Clinical Application in Select Solid Tumor Malignancies

Radiation therapy is a frequently used modality for the treatment of solid cancers. Although the mechanisms of cell kill are similar for all forms of radiation, the in vivo properties of photon and proton beams differ greatly and maybe exploited to optimize clinical outcomes. In particular, proton particles lose energy in a predictable manner as they pass through the body. This property is used clinically to control the depth at which the proton beam is terminated, and to limit radiation dose beyond the target region. This strategy can allow for substantial reductions in radiation dose to normal tissues located just beyond a tumor target. However, the degradation of proton energy in the body remains highly sensitive to tissue density. As a consequence, any changes in tissue density during the course of treatment may significantly alter proton dosimetry. Such changes may occur through alterations in body weight, respiration, or bowel filling/gas, and may result in unfavorable dose deposition. In this manuscript, we provide a detailed method for the delivery of proton therapy using both passive scatter and pencil beam scanning techniques for prostate cancer. Although the described procedure directly pertains to prostate cancer patients, the method may be adapted and applied for the treatment of virtually all solid tumors. Our aim is to equip readers with a better understanding of proton therapy delivery and outcomes in order to facilitate the appropriate integration of this modality during cancer therapy.

The fundamentals of radiation planning and delivery for proton therapy using prostate cancer as a model are presented. The application of these principles to other selected disease sites highlights how proton radiotherapy may enhance clinical outcomes for cancer patients.

Radiation therapy is a frequently used modality for the treatment of solid cancers. Although the mechanisms of cell kill are similar for all forms of ...

Immunoglobulin G N-Glycan Analysis by Ultra-Performance Liquid Chromatography

Glycomics is a new subspecialty in omics system research that offers significant potential in discovering next-generation biomarkers for disease susceptibility, drug target discovery, and precision medicine. Alternative IgG N-glycans have been reported in several common chronic diseases and suggested to have great potential in clinical applications (i.e., biomarkers for diagnosis and prediction of diseases). IgG N-glycans are widely characterized using the method of hydrophilic interaction chromatography (HILIC) ultra-performance liquid chromatography (UPLC). UPLC is a stable detection technology with good reproducibility and high relative quantitative accuracy. In addition, the structure of IgG N-glycan is clearly separated, and glycan composition and relative abundance in plasma are characterized.

Immunoglobulin G (IgG) N-glycan is characterized using hydrophilic interaction chromatography UPLC. In addition, the structure of IgG N-glycan is clearly separated. Presented here is an introduction to this experimental method so that it can be widely used in research settings.

Glycomics is a new subspecialty in omics system research that offers significant potential in discovering next-generation biomarkers for disease ...

Application of Lucilia sericata Larvae in Debridement of Pressure Wounds in Outpatient Settings

Biological therapy using Lucilia sericata larvae has numerous advocates worldwide, yet it is still fairly unknown and not commonly applied in daily practice because of the limited awareness and insufficient experience of medical and nursing personnel. There are case reports suggesting that maggot therapy can be applied and supported by lay caregivers, provided they are supervised and informed by physicians/nurses. The foregoing observation suggests that the method should be considered for implementation by a wider group of caregivers if accepted and meticulously supervised by trained and experienced medical staff. The concerns related to the therapeutic use of maggots in certain regions seem understandable, but are not supported by scientific facts. It should be noted that many therapeutic agents (including brood) used in medicine are of natural origin, and are associated with low production costs and high possibilities of implementation in the course of therapy. By analyzing the literature and using our own clinical and research experience, we have come to conclusions related to using larvae therapy, as a quick and safe method providing cleaning and revitalization in the process of treating wounds of various etiologies, especially pressure ulcers. In the current study, medical-grade Lucilia sericata maggots were applied to remove necrotic tissue from deep pressure sores. The treatment is mostly accepted by both caregivers and patients. In most cases, it is conducted by trained and experienced medical personnel in home and outpatient settings. Over the course of the conducted analyzes involving the collected specimens, there were no statistically significant relationships (p&gt; 0.05) confirmed between the wound surface successfully cleared by brood and variables, such as time from wound formation, location, surface size, and the depth of damage to the tissue structure. The lack of statistical dependence may result from the small size of the studied group. Based on the current findings, we have formulated the following conclusions: Maggot Debridement Therapy (MDT) is a fast and effective method enabling the preparation of the wound bed. The use of MDT in outpatient settings is safe and acceptable for patients and their caregivers.

Application of Lucilia sericata Larvae in Debridement of Pressure Wounds in Outpatient Settings

Maggots of Lucilia sericata were used in the debridement of deep pressure sores in outpatient settings for the treatment of pressure sores involving the complete skin thickness. The technique presented in the article does not pose risks for the patient.

Biological therapy using Lucilia sericata larvae has numerous advocates worldwide, yet it is still fairly unknown and not commonly applied in daily ...

Neuronavigation and Laparoscopy Guided Ventriculoperitoneal Shunt Insertion for the Treatment of Hydrocephalus

Hydrocephalus is a common adult neurosurgical condition typically requiring treatment with a cerebrospinal fluid (CSF) shunt, of which the ventriculoperitoneal (VP) shunt is the most common type. Unfortunately, the failure rates of VP shunts are alarmingly high, with up to 50% of patients requiring revision surgery within 2 years. VP shunt failure may occur due to infection, or catheter mispositioning, migration, and occlusion. We undertook a joint neurosurgery and general surgery collaboration in a 7-year prospective non-randomized consecutive quality improvement cohort study to reduce the rates of ventriculoperitoneal (VP) shunt failures in 224 adult patients at a tertiary care institution. The initiative combined the use of electromagnetic stereotactic neuronavigation to guide the placement of the proximal catheter and laparoscopy to place the distal catheter under direct visualization. With laparoscopic assistance, the distal catheter was anchored through a small hole created in the falciform ligament and placed into the right retrohepatic space, free from the omentum, adhesions, or bowel that might obstruct the catheter tip. The surgeries were performed using a shunt infection prevention protocol to reduce the risk of shunt infections. Here, we present an intraoperative video of the surgical procedure. Compliance with shunt infection reduction strategies and the combined utilization of neuronavigation and laparoscopy techniques in adult VP shunt surgery resulted in a 44% reduction in the risk of overall shunt failure. The significant positive impact with regard to shunt-failure-free patient outcomes among patients who underwent VP shunt surgery using this strategy underscores the value associated with the use of these modern intraoperative techniques and cross-specialty collaboration during VP shunt surgery.

Patient outcomes of ventriculoperitoneal (VP) shunt surgery, the mainstay treatment for hydrocephalus in adults, are poor due to high shunt failure rates. We present intraoperative footage of VP shunt insertion using neuronavigation and laparoscopy guidance, with the goal to reduce the risks of proximal and distal shunt catheter failures, respectively.

Hydrocephalus is a common adult neurosurgical condition typically requiring treatment with a cerebrospinal fluid (CSF) shunt, of which the ...

Polytetrafluoroethylene (PTFE) as a Suture Material in Tendon Surgery

With the evolution of suture materials, there has been a change in paradigms in primary and secondary tendon repair. Improved mechanical properties allow more aggressive rehabilitation and earlier recovery. However, for the repair to hold against higher mechanical demands, more advanced suturing and knotting techniques must be assessed in combination with those materials. In this protocol, the use of polytetrafluoroethylene (PTFE) as a suture material in combination with different repair techniques was investigated. In the first part of the protocol, both linear tension strength and elongation of knotted against not-knotted strands of three different materials used in flexor tendon repair were evaluated. The three different materials are polypropylene (PPL), ultra-high molecular weight polyethylene with a braided jacket of polyester (UHMWPE), and polytetrafluoroethylene (PTFE). In the next part (ex vivo experiments with cadaveric flexor tendons), the behavior of PTFE using different suture techniques was assessed and compared with PPL and UHMWPE.
This experiment is comprised of four steps: harvesting of the flexor tendons from fresh cadaveric hands, transection of the tendons in a standardized manner, tendon repair by four different techniques, mounting, and measurement of the tendon repairs on a standard linear dynamometer. The UHMWPE and PTFE showed comparable mechanical properties and were significantly superior to PPL in terms of linear traction strength. Repairs with four- and six-strand techniques proved stronger than two-strand techniques. Handling and knotting of PTFE are a challenge due to very low surface friction but fastening of the four- or six-strand repair is comparatively easy to achieve. Surgeons routinely use PTFE suture material in cardiovascular surgery and breast surgery. The PTFE strands are suitable for use in tendon surgery, providing a robust tendon repair so that early active motion regimens for rehabilitation can be applied.

Polytetrafluoroethylene PTFE as a Suture Material in Tendon Surgery

The present protocol illustrates a method for assessing the biophysical properties of tendon repairs ex vivo. A polytetrafluoroethylene (PTFE) suture material was evaluated by this method and compared to other materials under different conditions.

With the evolution of suture materials, there has been a change in paradigms in primary and secondary tendon repair. Improved mechanical properties allow ...