The authors would like to thank the second affiliated hospital of KMMU for providing cases and medical records related to this work. There is no funding support for this study.

The surgical method described in this paper has been approved by the Ethics Committee of the Second Affiliated Hospital of Kunming Medical University, and the use of patient surgical videos has been authorized. Informed consent was obtained from the patients, and patient data was used for presentation.
1. Instruments for operation
<ol>
	<li>Ensure that the display camera system and holmium laser are working properly.</li>
</ol>
2. Preparation for operation
<ol>
	<li>Use the following inclusion criteria to select participants for this study.
	<ol>
		<li>All patients who had been diagnosed with azoospermia or oligospermia and had consented to surgery.</li>
		<li>Semen volume less than 1.5 mL, pH&#60;7.2. Seminal plasma fructose &#8804;13 &#181;M/single ejaculation.</li>
		<li>Magnetic resonance imaging (MRI) revealed cysts in the ejaculatory duct area and enlarged bilateral seminal vesicle glands (Figure 1). 
		NOTE: It is difficult to identify prostate cysts and ejaculatory duct cysts through imaging accurately. We usually confirm the diagnosis through surgery when seminal vesicle dilatation has a width greater than 17 mm10.</li>
		<li>Sex hormone levels are within the normal range. The testicular volume was more than 15 mL (individual testicular volume). 
		NOTE: The purpose of testicular volume &#62;15 mL is to rule out azoospermia caused by some testicular factors. Testicular volume was measured using ultrasound report data and Lambert&#39;s formula: L x W x H x 0.71.</li>
		<li>There were no diseases that affected the safety of surgery, such as poorly controlled hypertension and hyperglycemia, respiratory diseases, etc.</li>
	</ol>
	</li>
	<li>Use the following exclusion criteria to exclude participants from this study.
	<ol>
		<li>Patients with urethral stricture, acute urinary tract infection, severe coagulopathy, or other contraindications to anesthesia.</li>
		<li>The testicular volume was less than 12 mL.</li>
	</ol>
	</li>
	<li>Administer intravenous antibiotics 30 min before surgery to prevent infection. Typically, use 1.5 g cefuroxime sodium with 100 mL of 0.9% sodium chloride solution.</li>
</ol>
3. Operational procedure
<ol>
	<li>Position the patient in lithotomy for anesthesia induction using sufentanil (0.3 &#181;g/kg), propofol (2 mg/kg), and rocuronium bromide (0.8 mg/kg). Perform tracheal intubation once drugs take effect, connecting to the anesthesia machine for mechanical ventilation.</li>
	<li>Maintain with propofol (2 mg/kg/h), remifentanil (0.15 &#181;g/kg/min), and 2% sevoflurane to achieve a BIS value of 40-60. Inject rocuronium bromide intermittently for inotropic relaxation. Disinfect the lower abdomen and perineal area 3x with iodophor.</li>
	<li>Connect the seminal vesiculoscope to the display system. Insert the endoscope through the external urethral opening and carefully advance it toward the posterior urethra. The urethral mucosa appears smooth and reddish, confirming the accurate positioning of the urethra within the visual field and extending towards the posterior urethra.</li>
	<li>If successful, check that the raised structure of the urethra, known as the colliculus seminalis, is visible.On either side of the colliculus seminalis, check for two small openings of the ejaculatory ducts (Figure 2A).</li>
	<li>Guide a wire (Bard) through the affected side&#39;s ejaculatory duct opening, entering the ejaculatory duct cyst. Observe a significant area filled with cloudy fluid during the procedure (Figure 2B).</li>
	<li>Flush the cyst with saline until achieving clear vision, examining for abnormal openings in the seminal canal. Enter the ipsilateral seminal vesicle through the abnormal ejaculatory duct cyst opening, revealing multiple honeycombed ductal lumens (Figure 2C).</li>
	<li>Using a 40 W holmium laser, expand the diameter of the ejaculatory tube to approximately 5 mm, facilitating improved circulation of the flush fluid and providing a clearer view (Figure 2D).</li>
	<li>Cut the ejaculatory duct opening along the direction of urine flow, which may be conducive to preserving the anti-urine reflux mechanism.</li>
	<li>Identify and explore the symmetrical position of the abnormal ejaculatory duct opening on the affected side.</li>
	<li>Enter the seminal vesicle of the healthy side for further exploration.Gently insert it into the seminal vesicle at the lower left and right of the cysts. It is important to note that the exact location may vary (Figure 2E).</li>
	<li>Use the holmium laser to precisely incise and widen the contralateral artificial opening, allowing smooth passage for the endoscope. This marks the pivotal endpoint of the operation, where sperm in the vas deferens on both sides is effectively released through the opening of the ejaculatory duct on the affected side (Figure 2F).</li>
	<li>Indwell a Fr18 catheter and rinse continuously with physiological sodium chloride to prevent blood clots from blocking the catheter.</li>
</ol>
4. Postoperative care
<ol>
	<li>On the 1st day post-surgery, continuously irrigate the bladder for 6 h and supplement with glucose and electrolytes as needed.</li>
	<li>Remove the catheter on the 2nd day post-surgery. Ensure that the patient experiences no evident pain after surgery; instead, they are able to endure the primary discomfort from the catheter, which is manageable without medication. Do not administer antibiotics postoperatively to patients without pre-existing infections. Keep the hospital stay for 2 days.</li>
	<li>Instruct the patients to increase water intake, urinate frequently, and ejaculate as soon as possible after discharge, aiming for 1-2 times per week. Additionally, closely observe the semen volume.</li>
</ol>
5. Follow up
<ol>
	<li>Perform semen analysis in the 3rd month after surgery to ensure that the gag time requirement was met, and follow up after 3 months mainly through telephone interviews.</li>
	<li>Perform semen samples using the automatic sperm motility testing instrument and the automatic seminal plasma biochemical instrument. Compare the semen analysis results obtained before the surgery with those at the 3rd month post-surgery.</li>
</ol>
6. Statistical analysis
<ol>
	<li>Analyze the results using GraphPad software and expressed as mean &#177; SD. Normality tests were performed using Shapiro-Wilk tests.If the preoperative and postoperative differences are normally distributed, analyze using the paired t-test; otherwise, use non-parametric tests (Mann-Whitney). Analyze differences between preoperative and postoperative data by paired t-test. Statistical differences were regarded as significant when P &#60;0.05.</li>
</ol>

Seminal Vesiculoscope Laser-Assisted Surgery to Treat Ejaculatory Duct Blockage

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	<li>Avellino, G. J., Lipshultz, L. I., Sigman, M., Hwang, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transurethral+resection+of+the+ejaculatory+ducts:+etiology+of+obstruction+and+surgical+treatment+options.">Transurethral resection of the ejaculatory ducts: etiology of obstruction and surgical treatment options.</a> Fertil Steril. 111 (3), 427-443 (2019).</li><li>Modgil, V., Rai, S., Ralph, D. J., Muneer, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+update+on+the+diagnosis+and+management+of+ejaculatory+duct+obstruction.">An update on the diagnosis and management of ejaculatory duct obstruction.</a> Nat Rev Urol. 13 (1), 13-20 (2016).</li><li>Paick, J., Kim, S. H., Kim, S. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ejaculatory+duct+obstruction+in+infertile+men.">Ejaculatory duct obstruction in infertile men.</a> BJU Int. 85 (6), 720-724 (2000).</li><li>Wang, 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=Transurethral+seminal+vesiculoscopy+using+a+6F+vesiculoscope+for+ejaculatory+duct+obstruction:+initial+experience.">Transurethral seminal vesiculoscopy using a 6F vesiculoscope for ejaculatory duct obstruction: initial experience.</a> J Androl. 33 (4), 637-643 (2012).</li><li>Galloway, M., Woods, R., Nicholson, S., Foggin, J., Elliott, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+audit+of+waiting+times+in+a+haematology+clinic+before+and+after+the+introduction+of+point-of-care+testing.">An audit of waiting times in a haematology clinic before and after the introduction of point-of-care testing.</a> Clin Lab Haematol. 21 (3), 201-205 (1999).</li><li>WD, Z., 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+effect+analysis+of+two+surgical+methods+for+the+treatment+of+ejaculatory+duct+obstruction.">Clinical effect analysis of two surgical methods for the treatment of ejaculatory duct obstruction.</a> Chinese J Androl. 4, 43-45 (2013).</li><li>Halpern, E. 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C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+and+treatment+of+ejaculatory+duct+obstruction+in+infertile+men.">Evaluation and treatment of ejaculatory duct obstruction in infertile men.</a> Can J Urol. 13 (Suppl 1), 18-21 (2006).</li><li>Tu, X. A., 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=Transurethral+bipolar+plasma+kinetic+resection+of+ejaculatory+duct+for+treatment+of+ejaculatory+duct+obstruction.">Transurethral bipolar plasma kinetic resection of ejaculatory duct for treatment of ejaculatory duct obstruction.</a> J Xray Sci Technol. 21 (2), 293-302 (2013).</li><li>El-Assmy, A., El-Tholoth, H., Abouelkheir, R. T., Abou-El-Ghar, M. 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The authors declare that they have no competing interests.

TURED is a primary surgical approach to treat ejaculatory duct obstruction caused by the ejaculatory duct cyst, and its main operation mode is to reveal the ejaculatory duct opening after the excision of cysts with an electric incision to relieve the pressure and dreg the seminal canal11. The study subjects were asked about their medical history during the visit, and all had normal sexual activity frequency (1-2 times per week) and no sexual dysfunction or hematospermia. The preoperative semen exa...

Ejaculatory duct obstruction is a rare disease of the male reproductive system, with a reported incidence of 1%-5%1,2. Ejaculatory duct cysts represent the predominant cause of ejaculatory duct obstruction. Semen examination in typical EDO patients reveals four distinctive characteristics: 1. Semen volume less than 2 mL, with a direct correlation between obstruction severity and decreased volume; 2. Oligospermia, with bilateral complete obstruction resulting in azoospermia; 3. Decreased pH value of semen; 4. Reduced levels of seminal plasma fructose, sometimes even dropping to 0 mM/L3. Male infertility caused by EDO can be treated with surgery and is less effective with conservative treatment4. In the past, the main method was transurethral resection of the ejaculatory duct. Although this approach boasts benefits like reduced trauma and fewer intraoperative complications, the surgical removal of the verumontanum disrupts the normal physiological structure of the distal ejaculatory duct. This, in turn, increases the postoperative risk of complications such as urinary reflux, epididymitis, retrograde ejaculation, and urinary incontinence5. At the same time, heat production during the operation may lead to the injury of the ejaculatory duct, seminal vesicle, and even rectum, and the thermal effect of the electric incision may cause new obstruction6.
The verumontanum stands as a crucial anatomical element within the male reproductive system, ensuring the precise and regulated discharge of semen during ejaculation while also helping to prevent retrograde flow. Whether the disadvantages of TURED can be ameliorated by preserving the seminal caruncle is unclear. Several studies have attempted to utilize laser-assisted endoscopy for the treatment of EDO while preserving verumontanum2,7,8,9. Although the surgical approaches varied, post-operative semen recovery was notably successful with minimal complications. This indicates that preserving the epididymal head may be beneficial. However, the method they used is relatively complex and does not intervene in the healthy ejaculatory duct, which may increase the risk of recurrence. Therefore, we present a simple and effective surgical method.
In this study, the seminal vesiculoscope was guided into the ejaculatory duct cyst on the affected side by a wire guide. Then, the holmium laser was used to enlarge the ejaculatory duct opening on the affected side to ensure that it had a sufficiently large outflow channel.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Camera system</td>
			<td>Karl Storz, Germany</td>
			<td>TC200EN</td>
			<td>Endoscopic camera system</td>
		</tr>
		<tr>
			<td>Fr18 Cathete</td>
			<td>Zhanjiang City Shida Industrial Co., Ltd.</td>
			<td>2660476</td>
			<td>Drainage of urine</td>
		</tr>
		<tr>
			<td>Fr6/7.5 vesiculoscope</td>
			<td>Richard Wolf, germany</td>
			<td>8702.534</td>
			<td>Operative procedure</td>
		</tr>
		<tr>
			<td>iodophor</td>
			<td>Shanghai Likang Disinfectant Hi-Tech Co., Ltd.</td>
			<td>31005102</td>
			<td>Skin disinfection</td>
		</tr>
		<tr>
			<td>Nitinol Guidewire 0.035&#34;</td>
			<td>C. R Bard, Inc. Covington, GA&#160;</td>
			<td>150NFS35</td>
			<td>Guide</td>
		</tr>
		<tr>
			<td>Propofol</td>
			<td>Sichuan Kelun Pharmaceutical Research Institute Co., Ltd.</td>
			<td>H20203571</td>
			<td>Induction and maintenance of anesthesia</td>
		</tr>
		<tr>
			<td>Remifentanil</td>
			<td>Yichang Humanwell Pharmaceuticals CO,Ltd.</td>
			<td>H20030200</td>
			<td>Maintenance of anesthesia</td>
		</tr>
		<tr>
			<td>Rocuronium bromide</td>
			<td>Zhejiang Huahai&#160; Pharmaceuticals CO,Ltd.</td>
			<td>H20183264</td>
			<td>Induction and maintenance of anesthesia</td>
		</tr>
		<tr>
			<td>Sevoflurane</td>
			<td>Jiangsu Hengrui Pharmaceuticals Co., Ltd.</td>
			<td>H20070172</td>
			<td>Maintenance of anesthesia</td>
		</tr>
		<tr>
			<td>Slimline EZ 200</td>
			<td>LUMENIS, USA</td>
			<td>0642-393-01</td>
			<td>Dissect capsule wall</td>
		</tr>
		<tr>
			<td>Sodium Chloride Physiological Solution</td>
			<td>Hua Ren MEDICAL TECHNOLOGY CO. Ltd.</td>
			<td>H20034093</td>
			<td>Flushing fluid</td>
		</tr>
		<tr>
			<td>Sufentanil</td>
			<td>Yichang Humanwell Pharmaceuticals CO,Ltd.</td>
			<td>H20054171</td>
			<td>Induction and maintenance of anesthesia</td>
		</tr>
		<tr>
			<td>Syringe 50 mL&#160;</td>
			<td>Double Pigeon Group Co. Ltd.</td>
			<td>20163141179</td>
			<td>Inject 0.9% sodium chloride solution into the vesiculoscope</td>
		</tr>
		<tr>
			<td>VersaPulse PowerSuite 100W Laser System</td>
			<td>LUMENIS, Germany</td>
			<td>PS.INT.100W</td>
			<td>Provide energy</td>
		</tr>
	</tbody>
</table>

treatment of ejaculatory duct obstruction by seminal vesiculoscopy assisted flow modification

Transurethral resection of ejaculatory duct (TURED) is a primary surgical approach to treat ejaculatory duct obstruction (EDO) caused by the ejaculatory duct cyst. Intraoperative excision of the verumontanum is usually required to expose the ejaculatory ducts. However, preserving the verumontanum structure allows for a better simulation of normal physiological anatomy. Maintaining the verumontanum may increase the risk of postoperative distal ejaculatory duct scarring, leading to recurrent obstruction or reduced semen volume. Therefore, we attempted a novel technique that preserves the verumontanum, which is relatively easier and safer compared to TURED. The following were the procedural steps: 1. A 6F seminal vesiculoscope was introduced through the external urethral orifice to the vicinity of the verumontanum, locating the opening of the affected-side ejaculatory duct and introducing a guidewire into the cyst. This successful step preserved the verumontanum, maximizing the retention of the anti-reflux mechanism in the distal ejaculatory duct. 2. The holmium laser enlarged the affected-side ejaculatory duct opening to 5 mm, decreasing the likelihood of postoperative closure of the ejaculatory duct opening and simplifying the procedure. 3. A window was created within the cyst to access the contralateral seminal vesicle, and then a holmium laser was used to burn and dilate the opening to 5 mm, redirecting the contralateral ejaculatory duct into the cystic cavity. This modification preserved the opening of the healthy-side ejaculatory duct and provided a new outflow passage for semen, reducing the risk of decreased semen volume postoperatively. The patients experienced no complications postoperatively, had shorter hospital stays, and showed improvement in semen volume. Hence, this surgical approach is simple yet effective.

A total of 5 patients were enrolled in this study, ranging in age from 27 to 34 years (median 31 years), with a disease course of 6 to 15 months (mean 9 months). Follow-up was 12 to 48 months (mean 24.8 months). All patients successfully completed the operation. The average operation time was 26 min and the average hospital stay was 2 days. All 5 cases had unilateral EDO and contralateral seminal vesicle dilatation. The demographic information, incorporating preoperative and postoperative data, pertaining to the patients...

Watch this Scientific Journal Video about Advanced Treatment of Seminal Duct Blockage Employing Endoscopy-Mediated Semen Channel Refluviation at JoVE.com

Author Spotlight: Advanced Treatment of Seminal Duct Blockage Employing Endoscopy-Mediated Semen Channel Refluviation 

This protocol introduces the clinical application of seminal vesicle endoscopy combined with holmium laser in the treatment of ejaculatory duct obstruction caused by ejaculatory duct cyst.

Treatment of Ejaculatory Duct Obstruction by Seminal Vesiculoscopy Assisted Flow Modification

Transurethral resection of ejaculatory duct (TURED) is a primary surgical approach to treat ejaculatory duct obstruction (EDO) caused by the ejaculatory ...

treatment-ejaculatory-duct-obstruction-seminal-vesiculoscopy-assisted

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2.1K Views.  The Second Affiliated Hospital of Kunming Medical University. This protocol introduces the clinical application of seminal vesicle endoscopy combined with holmium laser in the treatment of ejaculatory duct obstruction caused by ejaculatory duct cyst.

Video: Author Spotlight: Advanced Treatment of Seminal Duct Blockage Employing Endoscopy-Mediated Semen Channel Refluviation 

Cannulation of the Mouse Submandibular Salivary Gland via the Wharton's Duct

Severe salivary gland hypofunction is frequently found in patients with Sj&ouml;gren's syndrome and those who receiving therapeutic 
irradiation in their head and neck regions for cancer treatment. Both groups of patients experience symptoms such as xerostomia (dry mouth), dysphagia 
(impaired chewing and swallowing), severe dental caries, altered taste, oro-pharyngeal infections (candidiasis), mucositis, pain and discomfort.
One innovative approach of regenerative medicine for the treatment of salivary gland hypo-function is speculated in RS Redman, E Mezey 
et al. 2009: stem cells can be directly deposited by cannulation into the gland as a potent method in reviving the functions of the impaired organ. Presumably, 
the migrated foreign stem cells will differentiate into glandular cells to function as part of the host salivary gland. Also, this cannulation technique is an 
expedient and effective delivery method for clinical gene transfer application.
Here we illustrate the steps involved in performing the cannulation procedure on the mouse submandibular salivary gland via the Wharton's duct 
(Fig 1). C3H mice (Charles River, Montreal, QC, Canada) are used for this experiment, which have been kept under clean conventional conditions at 
the McGill University animal resource center. All experiments have been approved by the University Animal Care Committee and were in accordance with the 
guidelines of the Canadian Council on Animal Care.
For this experiment, a trypan blue solution is infused into the gland through the opening of the Wharton's duct using a 
insulin syringe with a 29-gauge needle encased inside a polyethylene tube. Subsequently, the mouse is dissected to show that the infusions migrated 
into the gland successfully.

Cannulation of the Mouse Submandibular Salivary Gland via the Wharton&#039;s Duct

A protocol for the cannulation of the mouse submandibular salivary gland via the Wharton's duct is described. For this experiment, the trypan blue solution is used as a dyer to demonstrate how this technique effectively delivers infusions into the targeted gland, and to suggest the reliability of this new approach as a potential clinical drug/cell therapy for the regeneration of salivary glands.

Severe salivary gland hypofunction is frequently found in patients with Sj&ouml;gren's syndrome and those who receiving therapeutic 
irradiation in their ...

Accuracy in Dental Medicine, A New Way to Measure Trueness and Precision

Reference scanners are used in dental medicine to verify a lot of procedures. The main interest is to verify impression methods as they serve as a base for dental restorations. The current limitation of many reference scanners is the lack of accuracy scanning large objects like full dental arches, or the limited possibility to assess detailed tooth surfaces. A new reference scanner, based on focus variation scanning technique, was evaluated with regards to highest local and general accuracy. A specific scanning protocol was tested to scan original tooth surface from dental impressions. Also, different model materials were verified. The results showed a high scanning accuracy of the reference scanner with a mean deviation of 5.3 &#177; 1.1 &#181;m for trueness and 1.6 &#177; 0.6 &#181;m for precision in case of full arch scans. Current dental impression methods showed much higher deviations (trueness: 20.4 &#177; 2.2 &#181;m, precision: 12.5 &#177; 2.5 &#181;m) than the internal scanning accuracy of the reference scanner. Smaller objects like single tooth surface can be scanned with an even higher accuracy, enabling the system to assess erosive and abrasive tooth surface loss. The reference scanner can be used to measure differences for a lot of dental research fields. The different magnification levels combined with a high local and general accuracy can be used to assess changes of single teeth or restorations up to full arch changes.

Accuracy is a major demand in dental medicine. To verify accuracy, reference scanners are needed. This article presents a new reference scanner with an adjusted scanning method to acquire a broad variety of dental morphologies with high trueness and precision.

Reference scanners are used in dental medicine to verify a lot of procedures. The main interest is to verify impression methods as they serve as a base ...

Bile Duct Ligation in Mice: Induction of Inflammatory Liver Injury and Fibrosis by Obstructive Cholestasis

In most vertebrates, the liver produces bile that is necessary to emulsify absorbed fats and enable the digestion of lipids in the small intestine as well as to excrete bilirubin and other metabolic products. In the liver, the experimental obstruction of the extrahepatic biliary system initiates a complex cascade of pathological events that leads to cholestasis and inflammation resulting in a strong fibrotic reaction originating from the periportal fields. Therefore, surgical ligation of the common bile duct has become the most commonly used model to induce obstructive cholestatic injury in rodents and to study the molecular and cellular events that underlie these pathophysiological mechanisms induced by inappropriate bile flow. In recent years, different surgical techniques have been described that either allow reconnection or reanastomosis after bile duct ligation (BDL), e.g., partial BDL, or other microsurgical methods for specific research questions. However, the most frequently used model is the complete obstruction of the common bile duct that induces a strong fibrotic response after 21 to 28 days. The mortality rate can be high due to infectious complications or technical inaccuracies. Here we provide a detailed surgical procedure for the BDL model in mice that induce a highly reproducible fibrotic response in accordance to the 3R rule for animal welfare postulated by Russel and Burch in 1959.

Disruption of bile flow results in severe inflammatory cholestatic liver injury with a characteristic time-dependent sequence of morphological alterations. Here we present a protocol for the surgical ligation of the common bile duct in mice that allows to induce a strong fibrotic response after 21 to 28 days.

In most vertebrates, the liver produces bile that is necessary to emulsify absorbed fats and enable the digestion of lipids in the small intestine as well ...

A Murine Model of Irreversible and Reversible Unilateral Ureteric Obstruction

Obstruction of the kidney may affect native or transplanted kidneys and results in kidney injury and scarring. Presented here is a model of obstructive nephropathy induced by unilateral ureteric obstruction (UUO), which can either be irreversible (UUO) or reversible (R-UUO). In the irreversible UUO model, the ureter may be obstructed for variable periods of time in order to induce increasingly severe renal inflammation and interstitial fibrotic scarring. In the reversible R-UUO model the ureter is obstructed to induce hydronephrosis, tubular dilation and inflammation. After a suitable period of time the ureteric obstruction is then surgically reversed by anastomosis of the severed previously obstructed ureter to the bladder in order to allow complete decompression of the kidney and restoration of urinary flow to the bladder. The irreversible UUO model has been used to investigate various aspects of renal inflammation and scarring including the pathogenesis of disease and the testing of potential anti-inflammatory or anti-fibrotic therapies. The more challenging model of R-UUO has been used by some investigators and does offer significant research potential as it allows the study of inflammatory and immune processes and tissue remodeling in an injured and scarred kidney following the removal of the injurious stimulus. As a result, the R-UUO model offers investigators the opportunity to explore the resolution of kidney inflammation together with key aspects of tissue repair. These experimental models are of relevance to human disease as patients often present with obstruction of the renal tract that requires decompression and are commonly left with significant residual kidney impairment that has no current treatment options and may lead to eventual end stage kidney failure.

The murine model of irreversible unilateral ureteric obstruction (UUO) is presented together with the model of reversible UUO in which the ureteric obstruction is relieved by anastomosis of the severed ureter into the bladder. These models enable the study of renal inflammation and scarring as well as tissue remodeling.

Obstruction of the kidney may affect native or transplanted kidneys and results in kidney injury and scarring. Presented here is a model of obstructive ...

Transcanalicular Diode Laser-assisted Dacryocystorhinostomy for the Treatment of Primary Acquired Nasolacrimal Duct Obstruction

Today&#39;s gold standard in the treatment of infrasaccal primary acquired nasolacrimal duct obstruction (PANDO) is external dacryocystorhinostomy (DCR), a relatively invasive procedure that can be performed after failure of recanalizing treatments. However, with progress in the field of diode laser technology, new approaches have emerged. Laser-assisted transcanalicular DCR with subsequent bicanalicular silicon intubation is a new option showing great promise as a viable minimally invasive procedure. Under permanent endoscopic visual control from the nasal cavity, a diode laser fiber is inserted into the lacrimal sac and laser energy is applied to create a bony ostium between the lacrimal sac and the nasal cavity. Since no skin incision needs to be made, advantages of this method comprise the sparing of the skin as well as the medial palpebral structures and the physiological palpebral-canalicular pump mechanism. The duration of surgery as well as reconvalescence is generally shorter than with external DCR. Complications include silicon tube prolapse, mild swelling and, rarely, canalicular infection and thermal injury. One-year functional success rates, defined as complete resolution of symptoms and ostium patency, are high, yet still range behind those of external DCR. However, secondary external DCR after failure of laser-assisted DCR can be performed without difficulty. Thus, laser-assisted transcanalicular DCR is a valid option that should be considered as a second-step procedure after failure of recanalization procedures and before external DCR.

The goal of this protocol is to present transcanalicular laser-assisted dacryocystorhinostomy as a minimally invasive approach in the treatment of primary acquired nasolacrimal duct obstruction.

Today&#39;s gold standard in the treatment of infrasaccal primary acquired nasolacrimal duct obstruction (PANDO) is external dacryocystorhinostomy (DCR), ...

OP-IVM: Combining In vitro Maturation after Oocyte Retrieval with Gynecological Surgery

The use of in vitro maturation (IVM) before gynecological operation (OP-IVM) is an extension of conventional IVM that combines IVM following oocyte retrieval with routine gynecological surgery. OP-IVM is suitable for patients undergoing benign gynecological surgery who have the need for fertility preservation (FP) or infertility treatments such as in vitro fertilization and embryo transfer (IVF-ET). In the operating room, patients undergoing benign gynecological surgery are first anesthetized and receive ultrasound-guided immature follicle aspiration (IMFA) treatment. As the subsequent gynecological surgery is performed, the cumulus-oocyte complexes (COCs) are examined, and the immature COCs are transferred into the IVM medium and cultured for 28-32 hours in the IVF laboratory. After assessment, mature oocytes in the MII stage will be selected and cryopreserved in liquid nitrogen for FP or fertilized by intracytoplasmic sperm injection (ICSI) for IVF-ET. By combining IVM with gynecological surgery, immature oocytes that would have been discarded can be saved and used for assisted reproductive technology (ART). The procedure, significance and critical aspects of OP-IVM are described in this article.

In vitro maturation (IVM) before gynecological operation (OP-IVM) combines IVM following oocyte retrieval with routine gynecological surgery and serves as an extension of conventional IVM applications for fertility preservation.

The use of in vitro maturation (IVM) before gynecological operation (OP-IVM) is an extension of conventional IVM that combines IVM following oocyte ...

Quality-Controlled Sputum Analysis by Flow Cytometry

Sputum, widely used to study the cellular content and other microenvironmental features to understand the health of the lung, is traditionally analyzed using cytology-based methodologies. Its utility is limited because reading the slides is time-consuming and requires highly specialized personnel. Moreover, extensive debris and the presence of too many squamous epithelial cells (SECs), or cheek cells, often renders a sample inadequate for diagnosis. In contrast, flow cytometry allows for high-throughput phenotyping of cellular populations while simultaneously excluding debris and SECs.
The protocol presented here describes an efficient method to dissociate sputum into a single cell suspension, antibody stain and fix cellular populations, and acquire samples on a flow cytometric platform. A gating strategy that describes the exclusion of debris, dead cells (including SECs) and cell doublets is presented here. Further, this work also explains how to analyze viable, single sputum cells based on a cluster of differentiation (CD)45 positive and negative populations to characterize hematopoietic and epithelial lineage subsets. A quality control measure is also provided by identifying lung-specific macrophages as evidence that a sample is derived from the lung and is not saliva. Finally, it has been demonstrated that this method can be applied to different cytometric platforms by providing sputum profiles from the same patient analyzed on three flow cytometers; Navios EX, LSR II, and Lyric. Furthermore, this protocol can be modified to include additional cellular markers of interest. A method to analyze an entire sputum sample on a flow cytometric platform is presented here that makes sputum amenable for developing high-throughput diagnostics of lung disease.

This protocol describes an efficient method for dissociating sputum into a single cell suspension and the subsequent characterization of cellular subsets on standard flow cytometric platforms.

Sputum, widely used to study the cellular content and other microenvironmental features to understand the health of the lung, is traditionally analyzed ...

DUCT: Double Resin Casting followed by Micro-Computed Tomography for 3D Liver Analysis

The liver is the biggest internal organ in humans and mice, and high auto-fluorescence presents a significant challenge for assessing the three-dimensional (3D) architecture of the organ at the whole-organ level. Liver architecture is characterized by multiple branching lumenized structures, which can be filled with resin, including vascular and biliary trees, establishing a highly stereotyped pattern in the otherwise hepatocyte-rich parenchyma. This protocol describes the pipeline for performing double resin casting micro-computed tomography, or &#34;DUCT&#34;. DUCT entails injecting the portal vein and common bile duct with two different radiopaque synthetic resins, followed by tissue fixation. Quality control by clearing one lobe, or the entire liver, with an optical clearing agent, allows for pre-screening of suitably injected samples. In the second part of the DUCT pipeline, a lobe or the whole liver can be used for micro-computed tomography (microCT) scanning, (semi-)automated segmentation, and 3D rendering of the portal venous and biliary networks. MicroCT results in 3D coordinate data for the two resins allowing for qualitative as well as quantitative analysis of the two systems and their spatial relationship. DUCT can be applied to postnatal and adult mouse liver and can be further extended to other tubular networks, for example, vascular networks and airways in the lungs.

Double resin casting micro-computed tomography, or DUCT, enables visualization, digitalization, and segmentation of two tubular systems simultaneously to facilitate 3D analysis of organ architecture. DUCT combines ex vivo injection of two radiopaque resins followed by micro-computed tomography scanning and segmentation of the tomographic data.

The liver is the biggest internal organ in humans and mice, and high auto-fluorescence presents a significant challenge for assessing the ...

A Modified Vessel-Sparing Microsurgical Vasoepididymostomy

Microsurgical vasoepididymostomy (MVE) is the main surgical treatment for epididymal obstruction. The vasal vessels are ligated during MVE. However, preserving the vasal vessels during MVE might better simulate the normal physiological structure and be meaningful for patients who have undergone varicocelectomy. Nevertheless, preserving the vasal vessels might elevate the risk of increasing the tension of anastomosis, affecting the patency rate and leading to delayed postoperative bleeding. Therefore, we developed a novel vessel-sparing MVE to make it safer. Here is a summary of the improvements to the procedure. 1) The retrograde dissociation of the vasal vessels on the proximal testicular side was adopted as the main method, and the anterograde dissociation of the vasal vessels on the distal testicular side was adopted as a supplement to dissociate the vasal vessels to be preserved. This improvement ensures the blood supply to the vas deferens that will be used for anastomosis and also provides longer vasal vessels, which reduces the tension of anastomosis. 2) By fixing the vas deferens to be anastomosed and the broken end of the vas deferens, the free vasal vessels get fixed, which resolves the problem of transmission of vas tension to the vasal vessels and reduces the risk of vasal vessel hemorrhage. 3) Dissociation of the vas deferens after opening the tunica vaginalis increases the mobilization of the vas deferens, which also makes the new procedure easier to complete. The evaluation of the outcomes of this new procedure showed that no significant postoperative complications occurred in the patients, and the patency rate was no different from that of the conventional procedure. Therefore, this new, improved procedure can be considered safe, with satisfactory postoperative results.

Here, we present a protocol to preserve the vasal vessels in microsurgical vasoepididymostomy. The surgical security is enhanced by preserving the vasal vessels using a retrograde-anterograde dissociation and fixing the vasal vessels.

Microsurgical vasoepididymostomy (MVE) is the main surgical treatment for epididymal obstruction. The vasal vessels are ligated during MVE. However, ...

Orthopedic Robot-Assisted Femoral Neck System in the Treatment of Femoral Neck Fracture

Cannulated screw fixation is the main therapy for femoral neck fractures, especially in young patients. The traditional surgical procedure uses C-arm fluoroscopy to place the screw freehand and requires several guide wire adjustments, which increases the operation time and radiation exposure. Repeated drilling can also cause damage to the blood supply and bone quality of the femoral neck, which can be followed by complications such as screw loosening, nonunion, and femoral head necrosis. In order to make fixation more precise and reduce the incidence of complications, our team applied robot-assisted orthopedic surgery for screw placement using the femoral neck system to modify the traditional procedure. This protocol introduces how to import a patient's X-ray information into the system, how to perform screw path planning in software, and how the robotic arm assists in screw placement. Using this method, the surgeons can place the screw successfully the first time, improve the accuracy of the procedure, and avoid radiation exposure. The whole protocol includes the diagnosis of femoral neck fracture; the collection of intraoperative X-ray images; screw path planning in the software; precise placement of the screw under the assistance of the robotic arm by the surgeon; and verification of the implant placement.

This article introduces a method of robot-assisted orthopedic surgery for screw placement during the treatment of femoral neck fracture using the femoral neck system, which allows for more accurate screw placement, improved surgical efficiency, and fewer complications.

Cannulated screw fixation is the main therapy for femoral neck fractures, especially in young patients. The traditional surgical procedure uses C-arm ...