Does the presence of blind-ended vas deferens and spermatic vessels in laparoscopic exploration of non-palpable testes conclusively indicate testicular absence?

Objective: The purpose of this study was to determine whether the presence of blind-ended vas deferens and spermatic vessels (VDSV) during laparoscopic exploration of non-palpable testes (NPT) indicates testicular absence or atrophy. Materials and methods: A retrospective analysis was conducted on clinical data of patients diagnosed with NPT and treated with surgical intervention at our center from April 2013-April 2023. The dataset encompassed information such as the children's age, affected side, size of the contralateral testis, surgical procedures employed, outcomes, and histopathological examination results. All patients underwent physical examination and ultrasonography preoperatively, followed by a combination of laparoscopic exploration and exploration through inguinal or scrotal incisions during surgery. Long-term follow-up was conducted postoperatively. Results: A total of 476 cases comprising 504 NPT were included in this study: 302 cases on the left side, 146 cases on the right side, and 28 cases bilaterally. All patients underwent surgical treatment within 6-126 months (median 13 months). During laparoscopic exploration, blind-ended VDSV were found in 90 testes (72 on the left side, 18 on the right side), while exploration through inguinal or scrotal incisions revealed 52 (57.8%) testicular nodules with atrophy, which were excised, leaving 38 (42.2%) without any findings. Histopathological examination of atrophic nodules revealed fibrosis as the most common finding in 41 cases (78.8%), followed by involvement of the vas deferens in 33 cases (63.5%), calcification in 24 cases (46.2%), epididymis in 23 cases (44.2%), and hemosiderin deposition in 7 cases (13.6%). Fibrosis, calcification, hemosiderin deposition, involvement of the vas deferens, and epididymis were found in combination in 47 specimens (90.4%). Seminiferous tubules (SNT) were found in 3 specimens (5.7%), and germ cells (GC) were found in 1 specimen (1.9%). Conclusion: The presence of blind-ended VDSV during laparoscopic exploration of NPT does not necessarily indicate testicular absence or disappearance. It is possible that atrophic testicular nodules are located within the inguinal canal or scrotum. This understanding contributes to the management of non-palpable testes. Considering their unpredictable malignant potential, we recommend excision.

Efficacy comparison between Mathieu combined urethral plate incision and onlay island flap urethroplasty for distal hypospadias in patients with urethral plate stenosis.

The aim of the study was to compare the efficacy of Mathieu combined urethral plate incision (Mathieu-IP) versus onlay island flap urethroplasty in patients with distal hypospadias complicated by urethral stenosis. The clinical data of 70 patients with distal hypospadias complicated by urethral plate stenosis treated in the Department of Urology, Anhui Provincial Children's Hospital (Hefei, China), from May 2019 to May 2022, were retrospectively analyzed. Thirty-eight patients underwent Mathieu-IP (Mathieu-IP group) and 32 underwent onlay island flap urethroplasty (Onlay group). Follow-ups at 1 month, 6 months, and 12 months postoperatively assessed operative time, complications, urethral meatus morphology, and family satisfaction. The Mathieu-IP group had significantly shorter operative time (mean ± standard deviation [s.d.]: 81.58 ± 5.18 min) versus the Onlay group (mean ± s.d.: 110.75 ± 6.05 min; P < 0.05). Surgical success rates were 78.9% (Mathieu-IP group) and 75.0% (Onlay group), with no significant difference ( P > 0.05). Complications were comparable between the groups. The Mathieu-IP group resulted in a vertical slit-shaped urethral meatus in 89.5% versus 13.8% in the Onlay group ( P < 0.05). Family satisfaction with general penile appearance and skin shape showed no significant differences, but the Mathieu-IP group had higher satisfaction with meatal position ( P < 0.05). Mathieu-IP offers simplicity, safety, and shorter operative time compared to Onlay. Both the techniques effectively treat urethral plate stenosis in distal hypospadias, with reduced postoperative complications compared to tubularized incised plate urethroplasty. Mathieu-IP results in a vertical slit-shaped urethral meatus which enhances urinary stream, indicating its potential for broader adoption.

Management of pediatric vanishing testes syndrome based on pathological diagnosis: a single-center retrospective study.

This study aims to explore the optimal management strategy for pediatric vanishing testes syndrome (VTS) based on pathological characteristics. We retrospectively analyzed clinical data and pathological results of children with unilateral VTS who underwent surgical treatment at our center from July 2012 to July 2023. The children were categorized into the testicular excision group and testicular preservation group based on the surgical approach. Clinical characteristics and outcomes were compared between the two groups. Pathological examination results of excised testicular tissues were collected and analyzed, and long-term follow-up was conducted. A total of 368 children were included in this study. The age of the children at the time of surgery was 27 months (range, 6-156). Among them, 267 cases (72.6%) had VTS on the left side, and 101 cases (27.4%) on the right side. There were no statistically significant differences (P > 0.05) in age, affected side, contralateral testicular hypertrophy (CTH), testicular location, and preferred surgical incision between the testicular excision group (n = 336) and the testicular preservation group (n = 32). In the preservation group, two children experienced scrotal incision infections, showing a statistically significant difference compared to the excision group (P < 0.05). Pathological examination of excised tissues revealed fibrosis as the most common finding (79.5%), followed by vas deferens involvement (67%), epididymis involvement (40.5%), calcification (38.4%), and hemosiderin deposition (17.9%). Seminiferous tubules (SNT) was present in 24 cases (7.1%), germ cells (GC)in 15 cases (4.5%), and ectopic adrenal cortical tissue(EACT) in 1 case (0.3%). VTS belongs to a type of non-palpable testes (NPT) and requires surgical exploration. Considering the risk of scrotal incision infection after preserving atrophic testicular remnants and the unpredictable malignant potential, we recommend excision.

NRTPredictor: identifying rice root cell state in single-cell RNA-seq via ensemble learning.

BACKGROUND: Single-cell RNA sequencing (scRNA-seq) measurements of gene expression show great promise for studying the cellular heterogeneity of rice roots. How precisely annotating cell identity is a major unresolved problem in plant scRNA-seq analysis due to the inherent high dimensionality and sparsity. RESULTS: To address this challenge, we present NRTPredictor, an ensemble-learning system, to predict rice root cell stage and mine biomarkers through complete model interpretability. The performance of NRTPredictor was evaluated using a test dataset, with 98.01% accuracy and 95.45% recall. With the power of interpretability provided by NRTPredictor, our model recognizes 110 marker genes partially involved in phenylpropanoid biosynthesis. Expression patterns of rice root could be mapped by the above-mentioned candidate genes, showing the superiority of NRTPredictor. Integrated analysis of scRNA and bulk RNA-seq data revealed aberrant expression of Epidermis cell subpopulations in flooding, Pi, and salt stresses. CONCLUSION: Taken together, our results demonstrate that NRTPredictor is a useful tool for automated prediction of rice root cell stage and provides a valuable resource for deciphering the rice root cellular heterogeneity and the molecular mechanisms of flooding, Pi, and salt stresses. Based on the proposed model, a free webserver has been established, which is available at https://www.cgris.net/nrtp .

The diagnosis and treatment of testicular torsion in children with non-scrotal initial symptoms.

Objective: To explore the clinical characteristics of testicular torsion in children with non-scrotal initial symptoms who were misdiagnosed. Methods: A retrospective analysis of 73 cases children with testicular torsion and non-scrotal symptoms who were admitted to our department from October 2013 to December 2021 was performed. Patients were divided into misdiagnosis (27 cases) and clear diagnosis at first visit (46 cases) groups. Clinical data, including age at surgery, clinical presentation, physical examination, number of visits (≥2 times), affected side, time from initial symptoms to surgery, and surgical outcomes, were collected. The TWIST (Testicular Workup for Ischemia and Suspected Torsion) score was calculated and analyzed. Results: Statistically significant differences between the misdiagnosis and clear diagnosis groups were seen in the time from initial symptoms to surgery, the number of visits, the degree of testicular torsion, and the rate of orchiectomy (P < 0.05). There were no statistically significant differences (P > 0.05) in age, affected side, TWIST score, guardian, direction of testicular torsion, intra-vaginal or extra-vaginal torsion, and Arda classification. Postoperative follow-up was 6-40 months. Of the 36 patients who required an orchiopexy, 1 had testicular atrophy at six months and 2 were lost to follow-up. The contralateral testis of the 37 children who underwent orchiectomies developed normally without torsion. Conclusions: The clinical manifestations of testicular torsion in children are diverse and can easily lead to misdiagnosis. Guardians should be aware of this pathology and seek timely medical attention. When the initial diagnosis and treatment of testicular torsion is difficult, the TWIST score during the physical examination may be useful, especially for patients with intermediate-to-high risk scores. Color Doppler ultrasound can assist in making the diagnosis, but when testicular torsion is highly suspected, routine ultrasound is not necessary as it may lead to delayed surgical treatment.

A Single-Center Study of the Efficacy of Transvesicoscopic Ureterovesical Reimplantation: With or Without Ureteral Tailoring in Children with Congenital Megaureter.

Objective: To investigate the efficacy of ureteral tailoring with or without ureteral bladder reimplantation in the treatment of primary megaureter (MGU) in children by transvesicoscopic ureteral reimplantation. Methods: Clinical data from 21 children with primary MGU who were treated with pneumo-bladder ureterovesical reimplantation in Anhui Children's Hospital from January 2019 to August 2022 were retrospectively analyzed. Thereafter, children were divided into two groups according to whether intraoperative ureteral tailoring was performed, including 9 in the trimmed group and 12 in the nontrimmed group. In the meantime, there were 16 males and 5 females, with the age of 4.7 ± 2.12 years. Furthermore, there were 17 obstructive MGU cases, 2 reflux MGU cases, and 2 obstructive with reflux MGU cases, with 16 cases on the left side (including 1 case of Y-shaped ureter in the left duplicated kidney), whereas 5 on the right side. All children underwent transvesicoscopic ureteral reimplantation (the Cohen procedure). Thereafter, changes in operation time, duration of retained catheterization, hematuria time, renal pelvis and ureteral dilatation, and tortuosity between two groups were compared 3 months after Double-J tube removal. Results: All children were effectively treated with transvesicoscopic ureteral reimplantation, the operation time (p = 0.02, p < 0.05), postoperative hematuria time (p = 0.00, p < 0.05), and retained urinary catheter time (p = 0.00, p < 0.05) were shorter in the nontrimmed group, and the mean postoperative follow-up time was 27.6 (3-22) months. Moreover, the postoperative anteroposterior diameters of the affected renal pelvis in both groups were 0.31 ± 0.39 cm and 0.27 ± 0.29 cm, respectively, whereas the postoperative maximum ureteral diameters were 0.33 ± 0.31 cm and 0.27 ± 0.36 cm, separately, which were significantly reduced compared with the preoperative counterparts. All children recovered well, with no postoperative complications. Conclusion: Whether intraoperative ureteral clipping is performed or not does not significantly affect the outcome of transvesicoscopic laparoscopic ureteral bladder reimplantation for primary MGU in children. Both groups of children improved with satisfactory surgical results, but the nontrimmed group has shorter operation time, simpler operation, and less trauma.

Case report: Localized xanthogranulomatous pyelonephritis in children: A case report and literature review.

Background: Xanthogranulomatous pyelonephritis (XGPN), which is featured by inflammatory destruction of renal parenchyma and fibrosis of kidney, occurs mainly among adults, sporadically among children and rarely among infants. Recurrent urinary tract infections, kidney stone-induced obstructive nephropathy, malnutrition, abnormal lipid metabolism, hypoimmunity, lymphatic obstruction and congenital urinary abnormalities may all cause XGPN among children. Its primary treatment is radical nephrectomy. Case description: In this study, we describe a rare case of XGPN in a 7-year-old boy infected with Staphylococcus aureus (S. aureus). The child presented with symptoms including recurrent fever, urine culture negative. The postoperative pathology confirmed XGPN. Besides, partial nephrectomy was performed. Conclusion: XGPN, as a special type of chronic pyelonephritis, is a rare pyelonephritis requiring surgical treatment. Early diagnosis and treatment are crucial to reducing its morbidity and mortality. Although radical nephrectomy is the primary therapeutic option for patients with XGPN, partial nephrectomy surgery should be considered for focal XGPN, aiming to preserve residual renal function in children as far as possible.

A single-center study of two types of upper kidney preservation surgery for complete duplicated kidney in children.

Objective: The objectives of this study were to compare the efficacy, advantages, and disadvantages of insertable ureteral reimplantation (UC group) and ureteral end-to-side anastomosis (UU group) in the treatment of duplicated kidney and summarize the clinical experience in its diagnosis and treatment. Methods: The current retrospective study enrolled 20 cases with duplicated kidney in Anhui Provincial Children's Hospital from April 2016 to June 2021, including 11 in the UC group and 9 in the UU group. There were 8 boys and 12 girls, with 12 on the left side and 8 on the right side. Meanwhile, there were three cases with urinary tract infection and nine with urinary incontinence. The rest of them were found by B ultrasound during physical examination. The median age of these patients was 33.5 months. Later, preoperative and postoperative renal pelvis separation, ureteral dilation, operation time, and drainage tube indwelling time were compared between the two groups. Results: There were statistically significant differences in operation time (282 ± 50.55 vs. 176 ± 61.92, P = 0.03), drainage time (9.36 ± 5.00 vs. 5.33 ± 1.22, P = 0.02), and hospital stay (22.18 ± 5.40 vs. 14.78 ± 5.33, P = 0.007) between the two groups. In addition, the degree of hydronephrosis (UC: 1.86 ± 0.93 vs. 1.08 ± 0.77, P = 0.00; UU: 1.8 ± 0.95 vs. 0.89 ± 0.60, P = 0.02) and ureteral dilatation (UC: 1.57 ± 0.30 vs. 0.72 ± 0.22, P = 0.00; UU: 1.47 ± 0.50 vs. 0.88 ± 0.22, P = 0.001) were statistically different between the two groups before and after surgery. Conclusion: Compared with the UC method, the UU method has the advantages of less trauma, faster recovery, and fewer complications. Double J tube or ureter stent placement is beneficial for finding and protecting the lower ureter intraoperatively, without increasing the difficulty in operation, which can also prevent anastomosis or ureteral orifice stenosis.

Outcome analysis of early surgery and conservative treatment in neonates and infants with severe hydronephrosis.

OBJECTIVE: The treatment strategy and timing of ureteropelvic junction obstruction (UPJO) in infants remain controversial. This study aimed to compare the effect of early surgical treatment (EST) and conservative treatment (CT) on neonates and infants with UPJO and their recovery of renal function and morphology. METHODS: Eighty neonates and infants with severe hydronephrosis were enrolled in this study. They received early pyeloureteroplasty or CT. Diethylenetriamine pentaacetate was used to assess renal function. RESULTS: There were no significant differences in renal function or renal indices at baseline between the two groups. At 3 and 6 months of follow-up, the anteroposterior diameter of the renal pelvis and the Society of Fetal Urology grade in the EST surgery group were significantly lower compared with those at baseline. The thickness of the renal cortex was greater in the EST group than in the CT group at 3 and 6 months of follow-up. After follow-up for 6 months, renal function in the EST group was significantly better than that in the CT group. CONCLUSION: EST accelerates the recovery of renal morphological and functional indices in neonates and infants with severe hydronephrosis.

[Midline urethral-plate incision followed by Mathieu urethroplasty for repair of distal hypospadias].

OBJECTIVE: To compare the effect of midline urethral-plate incision followed by Mathieu urethroplasty (MUPI-MU) with that of tubularized incised-plate (TIP) urethroplasty in the treatment of distal hypospadias. METHODS: We retrospectively analyzed the clinical date on 72 cases of distal hypospadias treated in our hospital from August 2016 to January 2019, of which 21 (including 5 cases with a narrow urethral plate, small flat glans and shallow urethral groove) underwent MUPI-MU and the other 51 received TIP urethroplasty. We followed up the patients postoperatively and compared the shape and position of the urethral meatus and incidences of glanular dehiscence, fistula, stenosis and diverticulum between the two groups. RESULTS: After surgery, the urethral meatus was found vertical, slit-like and in a normal anatomical position in 19 cases (90.5%) in the MUPI-MU and 46 cases (90.2%) in the TIP group, with no statistically significant difference in the shape of the urethral meatus between the two groups (P>0.05). The postoperative incidence of fistula was significantly lower in the MUPI-MU than in the TIP group (1 ï¼»4.8%ï¼½ vs 15 ï¼»29.4%ï¼½, P = 0.048), and so was that of meatal stenosis (0 vs 12 ï¼»23.5%ï¼½, P = 0.037), but no statistically insignificant differences were observed between the MUPI-MU and TIP groups in the incidence of either glanular dehiscence (1 ï¼»4.8%ï¼½ vs 2 ï¼»9.8%ï¼½, P>0.05) or diverticulum (1 ï¼»4.8%ï¼½ vs 6 ï¼»11.8%ï¼½, P>0.05). No postoperative complications occurred in the 5 cases with a narrow urethral plate, small flat glans and shallow urethral groove. CONCLUSIONS: MUPI-MU can achieve a normal-looking, vertical, slit-like urethral meatus with a reasonable urethral diameter and minimized incidence of complications, especially applicable to the distal hypospadias patients with a narrow urethral plate, small flat glans and shallow urethral groove.

A chimeric recombinant infectious hematopoietic necrosis virus induces protective immune responses against infectious hematopoietic necrosis and infectious pancreatic necrosis in rainbow trout.

Infectious pancreatic necrosis virus (IPNV) and infectious hematopoietic necrosis virus (IHNV) are two common viral pathogens that cause severe economic losses in all salmonid species in culture, but especially in rainbow trout. Although vaccines against both diseases have been commercialized in some countries, no such vaccines are available for them in China. In this study, a recombinant virus was constructed using the IHNV U genogroup Blk94 virus as a backbone vector to express the antigenic gene, VP2, from IPNV via the reverse genetics system. The resulting recombinant virus (rBlk94-VP2) showed stable biological characteristics as confirmed by virus growth kinetic analyses, pathogenicity analyses, indirect immunofluorescence assays and western blotting. Rainbow trout were immunized with rBlk94-VP2 and then challenged with the IPNV ChRtm213 strain and the IHNV Sn1203 strain on day 45 post-vaccination. A significantly higher survival rate against IHNV was obtained in the rBlk94-VP2 group on day 45 post-vaccination (86%) compared with the PBS mock immunized group (2%). Additionally, IPNV loads decreased significantly in the rBlk94-VP2 immunized group in the liver (28.6-fold to 36.5-fold), anterior kidney (21.7-fold to 44.2-fold), and spleen (14.9-fold to 22.7-fold), as compared with the PBS mock control group. The mRNA transcripts for several innate and adaptive immune-related proteins (IFN-γ, IFN-1, Mx-1, CD4, CD8, IgM, and IgT) were also significantly upregulated after rBlk94-VP2 vaccination, and neutralizing antibodies against both IHNV and IPNV were induced on day 45 post-vaccination. Collectively, our results suggest that this recombinant virus could be developed as a vaccine vector to protect rainbow trout against two or more diseases, and our approach lays the foundations for developing live vaccines for rainbow trout.

Identification of the optimal insertion site for expression of a foreign gene in an infectious hematopoietic necrosis virus vector.

Infectious hematopoietic necrosis virus (IHNV) was developed as a vector to aid the construction of vaccines against viral diseases such as viral hemorrhagic septicemia virus, spring viremia of carp virus, and influenza virus H1N1. However, the optimal site for foreign gene expression in the IHNV vector has not been determined. In the present study, five recombinant viruses with the green fluorescence protein (GFP) gene inserted into different genomic junction regions of the IHNV genomic sequence were generated using reverse genetics technology. Viral growth was severely delayed when the GFP gene was inserted into the intergenic region between the N and P genes. Real-time fluorescence quantitative PCR assays showed that the closer the GFP gene was inserted towards the 3' end, the higher the GFP mRNA levels. Measurement of the GFP fluorescence intensity, which is the most direct method to determine the GFP protein expression level, showed that the highest GFP protein level was obtained when the gene was inserted into the intergenic region between the P and M genes. The results of this study suggest that the P and M gene junction region is the optimal site within the IHNV vector to express foreign genes, providing valuable information for the future development of live vector vaccines.

Vitamin D and Urinary Tract Infection: A Systematic Review and Meta-Analysis.

Whether or not Vitamin D deficiency is associated with urinary tract infections (UTI) remains controversial. We retrieved relevant articles from the PubMed, Google Scholar, the Cochrane Library, Med-line and Embase databases up to Mach 1, 2018 for studies investigating the association between Vitamin D and UTI. The meta-analysis of 9 studies included 1921 participants, of which 580 were diagnosed with UTI. They showed that Vitamin D insufficiency was associated with a significantly increased risk of having a UTI (pooled OR=3.01, 95%CI=2.31-3.91), with moderate heterogeneity (I2 =49.5%). Moreover, Vitamin D level was significantly lower in the UTI group (standardized mean difference (SMD)=-1.65, 95%CI=-2.69--0.60, P<0.001). Significant heterogeneity was also detected (I2 =97.9%, P<0.001). Meta-analysis also revealed a significant association between UTI and Vitamin D deficiency in children (OR=4.78, 95%CI=3.08-7.44, P<0.001). This meta-analysis indicated a significant association between Vitamin D insufficiency and increased risk of UTI, especially in children.

Recovery of recombinant infectious hematopoietic necrosis virus strain Sn1203 using the mammalian cell line BHK-21.

Reverse genetics systems are powerful tools for understanding the virulence mechanisms and gene functions of negative-sense RNA viruses. The reverse genetics systems commonly used for recombinant infectious hematopoietic necrosis virus (IHNV) are based on vaccinia virus infection. To avoid the potential biological safety risks associated with vaccinia virus, a recombinant IHNV virus strain Sn1203 (rIHNV-Sn1203) was rescued in this study using a mammalian cell line, BHK-21. The genome sequence authenticity of rIHNV-Sn1203 was confirmed using two silent genetic tags introduced by site-directed mutagenesis. Indirect immunofluorescence assays and transmission electron microscopy revealed that rIHNV-Sn1203 and wild-type IHNV-Sn1203 (wtIHNV-Sn1203) had identical immunogenicity and virion morphology. The virulence and pathogenicity of rIHNV-Sn1203 were assessed in vitro and in vivo. Although rIHNV-Sn1203 displayed trends toward delayed intracellular viral replication and lower virion yields compared with wtIHNV-Sn1203, statistical analyses revealed no significant differences between these two viruses. Moreover, rainbow trout challenged with rIHNV-Sn1203 and wtIHNV-Sn1203 showed indistinguishable mortality. Together, these results show that IHNV was successfully rescued using BHK-21 cells. This method is very convenient and may also be suitable for use in the recovery of other Novirhabdoviruses.

Kidney teratoma: A case report and literature review.

Teratomas are rare germline tumors that originate from embryonic germ cell layers. Teratoma of the kidney is extremely rare. We report the main radiologic features of an unusual case of mature cystic teratoma arising from the right kidney in a two-year-old boy. A left-sided abdominal mass was detected on physical examination and B-Mod Ultrasound (US) examination revealed a heterogeneous mass with central cystic component. Nephrectomy was performed and a large, fatty mass arising from the right kidney was excised. The final pathologic diagnosis was confirmed as cystic-solid renal teratoma.

Preliminary study of an oral vaccine against infectious hematopoietic necrosis virus using improved yeast surface display technology.

Infectious hematopoietic necrosis virus (IHNV) is a common pathogen that causes severe disease in the salmonid aquaculture industry. Because oral vaccines induce more efficient mucosal immunity than parenteral immunization, an oral vaccine was developed with an improved yeast cell surface display technology to induce an immune response to IHNV. The oral yeast vaccine, designated EBY100/pYD1-bi-G, was delivered orally to rainbow trout (Oncorhynchus mykiss) on days 1 and 32, and the nonspecific and specific immune responses were measured 50days after the first vaccination. In the hindgut, spleen, and head kidney, the expression of IFN-1 and Mx-1 was significantly upregulated after oral vaccination with EBY100/pYD1-bi-G, and the highest expression of IFN-1 and Mx-1 was observed in the spleen (7.5-fold higher than the control group) and head kidney (3.9-fold higher than the control group), respectively. Several markers of the adaptive immune response (IgM, IgT, CD4, and CD8) were also significantly upregulated, and the highest expression of these markers was observed in the hindgut, suggesting that the mucosal immune response was successfully induced by oral vaccination with EBY100/pYD1-bi-G. Sera from the orally vaccinated rainbow trout showed higher anti-IHNV neutralizing antibody titers (antibody titer 81±4) than the control sera (antibody titer 7±3), and the relative percentage survival after IHNV challenge was 45.8% compared with 2% in the control group. Although the protection afforded by this orally delivered vaccine was lower than that of a DNA vaccine (83%-98%), it is a promising candidate vaccine with which to protect larval fish against IHNV, which are most susceptible to the virus and difficult to inject with a DNA vaccine.

An efficient and simple method to increase the level of displayed protein on the yeast cell surface.

BACKGROUND: The development of oral vaccines using yeast surface display technology is an area of intensive study in vaccine development, but the protein level displayed on yeast surfaces is not currently high enough to obtain a robust immune response. METHODS: To address this issue, we established an efficient and simple method of increasing the level of displayed protein on the yeast cell surface. We used the single chain variable fragment (scFv) of an antibody against the infectious hematopoietic necrosis virus isolate Sn1203 as a target display protein. The yeast-derived scFv was first displayed on the yeast surface by galactose induction, and then Escherichia coli-derived scFv was also displayed on the same yeast via an artificial anchoring condition to increase the total scFv level on the yeast surface. RESULTS: The levels of yeast- and E. coli-derived scFv displayed on the yeast cell surface were analyzed by flow cytometry, western blotting, and fluorescent microscopy. The flow cytometry results indicated that when the cells were suspended in phosphate-buffered saline with 1mmol/L glutathione, 0.2mmol/L oxidized glutathione, and 5% dimethyl sulfoxide at 4°C for 6h, the E. coli-derived scFv protein was stably anchored to the yeast cell surface. The mean fluorescence intensity in these experiments, which is an indirect quantitative representation of the surface scFv expression, was three times higher in the treated cells than that in control cells. The western blotting results show two specific protein bands, the smaller of which was identified as the E. coli-derived scFv that was displayed on the yeast cell surface. Cell immunofluorescence is a more direct way to detect differentially produced proteins that are displayed on the yeast cell surface. The fluorescence microscopy results show that both fluorescence corresponding to the yeast-derived scFv and fluorescence corresponding to the E. coli-derived scFv can exist on the cell surface of same yeast cell. This confirms that the E. coli-derived scFv protein was successfully displayed on the yeast cell surface. CONCLUSIONS: This method provides a rapid, simple, and high-efficiency strategy to increase the level of displayed protein on the yeast cell surface. Application of this technique may allow the yeast surface display system to be used to generate potential oral vaccines.

[Traversing the vertical pedicle flap for the treatment of severely buried penis in children].

OBJECTIVE: To assess the effect of traversing the vertical pedicle flap (TVPF) for the treatment of severely buried penis in children. METHODS: Totally 43 children with severely buried penis underwentTVPF (n = 21)or modifiedShiraki surgery (control, n = 22) in our hospitalfrom February to December 2014. Wecompared the operation time, intra-operation blood loss, foreskin swelling time, and parents' satisfaction with penile appearance between the two groups. RESULTS: No statistically significant differences were observed between the TVPFand control groups in the operation time (ï¼»45.0 ±6.8ï¼½ vs ï¼»42.0 ±5.3ï¼½ min, P>0.05) or intra-operation blood loss(ï¼»5.0 ±1.2ï¼½ vs ï¼»6.0 ±0.8ï¼½ ml, P>0.05). The average foreskin swelling time was markedly shorter in the TVPFgroup than in the control (ï¼»9.0 ±2.3ï¼½ vs ï¼»15.0 ±4.8ï¼½ d, P<0.05)and the parents' satisfaction with penile appearancewas higher in the former than in the latter (95.23vs31.81 %, P<0.05). The elastic bandages were removedfor all the patientsat 3 days postoperatively, and 3 to 6-month follow-up revealed no penile retraction or relapse. CONCLUSIONS: The method of traversing the vertical pedicle flap is a feasible surgical option for the treatment of severely buried penis in children, which can make a rational use of the foreskin,remove the tight ring, and achieve a satisfactory appearance of the penis.

Recombinant scFv antibodies against infectious pancreatic necrosis virus isolated by flow cytometry.

Infectious pancreatic necrosis is a significant disease of farmed salmonids in China. In this study, a single chain variable fragment (scFv) antibody library derived from rainbow trout (Oncorhynchus mykiss) and viral protein VP2 of a Chinese infectious pancreatic necrosis virus (IPNV) isolate ChRtm213 were co-expressed by a bacterial display technology. The library was subjected to three rounds of screening by flow cytometry (FCM) to select IPNV specific antibodies. Six antibody clones with different mean fluorescence intensities (MFI) were obtained by picking colonies at random. The antibody clones were expressed and purified. The purified IPNV-specific scFv antibodies were used successfully in Western blotting, enzyme linked immunosorbent assay (ELISA) and an immunofluorescence antibody test (IFAT). This method provides a high throughput means to screen an antibody library by flow cytometry, and isolate a panel of antibody that can be used as potential reagents for the detection and study of IPNV that are prevalent in China.