Protocol for assessing myogenic tone and perfusion pressure in isolated mouse kidneys.

The isolated perfused kidney is a classic ex vivo preparation for studying renal physiology in general and vascular function. Here, we present a protocol for assessing myogenic tone in isolated mouse kidneys as well as vasodilatory and vasoconstrictive responses, expressed as perfusion pressure. We describe steps for pre-operative preparation, kidney and renal artery isolation, and connection of renal artery with glass cannula. We then detail how to measure pressure changes in perfused kidneys and the myogenic tone. For complete details on the use and execution of this protocol, please refer to Cui et al.1.

Ureteroscopic holmium laser to transect the greater omentum to remove an abdominal drain: Four case reports.

BACKGROUND: Drainage tube removal is difficult when the greater omentum becomes incarcerated in the drainage tube through the side holes. Currently, known removal methods are either ineffective or will cause additional damage to the patient in a secondary operation. Ureteroscopy and the holmium laser have been used in various surgical techniques in urology, and in theory, they are expected to be a good strategy for solving the problem of tissue incarceration. CASE SUMMARY: Four patients diagnosed with difficult removal of an abdominal drainage tube following abdominal surgery are reported. All patients underwent surgery to remove the incarcerated greater omentum in the drainage tube using a holmium laser and a ureteroscope, and a new 16-F drain was then placed in the abdominal or pelvic cavity. The efficacy of this technique was evaluated by intraoperative conditions, success rate, and operating time; safety was evaluated by perioperative conditions and the probability of postoperative complications. All four operations went smoothly, and the drains were successfully removed in all patients. The average operating time was 24.5 min. Intraoperatively, the average irrigation volume was 892.0 mL, the average drainage volume was 638.5 mL, and no bleeding or damage to surrounding tissues was observed. Postoperatively, the average drainage volume was 32.8 mL and the new drains were removed within 36 h. All patients were able to get out of bed and move around within 12 h. Their visual analogue pain scores were all below 3. The average follow-up duration was 12.5 mo and no complications such as fever or bleeding were noted. CONCLUSION: Ureteroscopic holmium laser surgery is an effective, safe and minimally invasive technique for removing drains where the greater omentum is incarcerated in the abdominal drain.

Contriving Multi-Epitope Subunit of Vaccine for COVID-19: Immunoinformatics Approaches.

COVID-19 has recently become the most serious threat to public health, and its prevalence has been increasing at an alarming rate. The incubation period for the virus is ~1-14 days and all age groups may be susceptible to a fatality rate of about 5.9%. COVID-19 is caused by a novel single-stranded, positive (+) sense RNA beta coronavirus. The development of a vaccine for SARS-CoV-2 is an urgent need worldwide. Immunoinformatics approaches are both cost-effective and convenient, as in silico predictions can reduce the number of experiments needed. In this study, with the aid of immunoinformatics tools, we tried to design a multi-epitope vaccine that can be used for the prevention and treatment of COVID-19. The epitopes were computed by using B cells, cytotoxic T lymphocytes (CTL), and helper T lymphocytes (HTL) base on the proteins of SARS-CoV-2. A vaccine was devised by fusing together the B cell, HTL, and CTL epitopes with linkers. To enhance the immunogenicity, the ß-defensin (45 mer) amino acid sequence, and pan-HLA DR binding epitopes (13aa) were adjoined to the N-terminal of the vaccine with the help of the EAAAK linker. To enable the intracellular delivery of the modeled vaccine, a TAT sequence (11aa) was appended to C-terminal. Linkers play vital roles in producing an extended conformation (flexibility), protein folding, and separation of functional domains, and therefore, make the protein structure more stable. The secondary and three-dimensional (3D) structure of the final vaccine was then predicted. Furthermore, the complex between the final vaccine and immune receptors (toll-like receptor-3 (TLR-3), major histocompatibility complex (MHC-I), and MHC-II) were evaluated by molecular docking. Lastly, to confirm the expression of the designed vaccine, the mRNA of the vaccine was enhanced with the aid of the Java Codon Adaptation Tool, and the secondary structure was generated from Mfold. Then we performed in silico cloning. The final vaccine requires experimental validation to determine its safety and efficacy in controlling SARS-CoV-2 infections.