[Robotic-assisted laparoscopic modified ventral onlay lingual mucosal graft in complex ureteral stricture construction: experience of eight cases].

Objective: To examine the efficacy of robot-assisted laparoscopic modified ventral onlay lingual mucosal graft for complex ureteral stricture. Methods: The clinical data of 8 patients with ureteral stricture admitted to the Department of Urology, General Hospital of Southern Theater Command from May to October 2022 were retrospectively analyzed. There were 6 males and 2 females, aged (45.1±10.2) years (range: 34 to 64 years), body mass index (24.6±2.0) kg/m2 (range: 20.7 to 26.6 kg/m2). Five cases on the left side, 3 cases on the right side, the length of the ureteral structure was (3.1±0.7) cm (range: 2.2 to 4.5 cm). The value of preoperative serum creatinine was (113.8±22.3) µmol/L (range: 96 to 15 µmol/L). Before excising the structure segment, the titched anastomosed part of the dorsal wall of the ureter, and then the posteriorly augmented anastomotic, the remaining ventral side was augmented with a onlay lingual mucosa graft, then the omentum flap was used to wrap the reconstructed ureteral segment. The lingual mucosa graft with a length of 2.5 to 5.0 cm and a width of 1.0 to 1.5 cm was cut according to the actual structure. The surgery information of the patient, complications, and recent follow-up were recorded. Results: The operation under robot-assisted laparoscopy was performed successfully in the 8 patients without conversion to open surgery. The duration of the operation was (226.9±22.8) minutes (range: 210 to 255 minutes), estimated blood loss was (93.8±25.9) ml (range: 75 to 150 ml), the retention time of the postoperative drainage tube was (4.8±1.3) days (range: 3 to 7 days), and the duration of postoperative hospitalization was (11.1±3.6) days (range: 9 to 14 days). One week after the operation, the patient could pronounce correctly, enunciate clearly, and eat normally. Double J tubes were removed 4 to 8 weeks after the operation. The follow-up time in this group was 3 to 9 months, the follow-up patients underwent imaging and other examinations, which showed a significant improvement in hydronephrosis on the affected side, and the value of renal pelvic separation on the affected side was (1.4±0.8) cm (range: 0 to 2.3 cm). The serum creatinine value was (100.1±24.9) µmol/L (range: 76 to 155 µmol/L). Three months after the operation, the ureteroscopy showed that the ureter was smooth and the mucosa was normal. Conclusions: Robot-assisted laparoscopic ureteroplasty with a lingual mucosal graft is a safe and feasible operation for complex ureteral stricture without serious complications, which provides a surgical option for repairing ureteral stricture.

[Clinical and immunological characteristics of patients with anti-synthetase syndrome complicated with cardiac involvement].

OBJECTIVE: To investigate the clinical and immunological features of cardiac involvement in patients with anti-synthetase syndrome (ASS). METHODS: In the study, 96 patients diagnosed with ASS hospitalized in the Department of Rheumatology and Immunology, Peking University People's Hospital from April 2003 to November 2020 were included. The patients were divided into two groups according to whether they were accompanied with cardiac involvement. Demographic features, clinical characteristics (Gottron's sign/papules, muscle damage, etc.), comorbidities, laboratory indices (creatine kinase, inflammatory indicators, immunoglobulin, complement, lymphocyte subset, autoantibodies, etc.) were collected and the differences between the two groups were analyzed statistically. RESULTS: The prevalence of cardiac involvement in the patients with ASS was 25.0% (24/96). The ASS patients complicated with cardiac involvement presented with elevated cardiac troponin I (cTnI, 75.0%, 18/24), pericardial effusion (33.3%, 8/24), reduction of left ventricular function (33.3%, 8/24) and valves regurgitation (33.3%, 8/24). The age of onset of the patients with cardiac involvement was older than that of the patients without cardiac involvement [(54.58±10.58) years vs. (48.47±13.22) years, P=0.043). Arthritis was observed less frequently in the patients with cardiac involvement than those without cardiac involvement (37.5% vs. 61.1%, P=0.044). In addition, rapidly progressive interstitial lung disease (54.2% vs. 30.6%, P=0.037) was observed more frequently in the patients with cardiac involvement than those without cardiac involvement. As compared with the ASS patients without cardiac involvement, C-reactive protein (CRP) [(13.55 (8.96, 38.35) mg/L vs. 4.60 (1.37, 17.40) mg/L, P=0.001], and lactate dehydrogenase (LDH) [408.0 (255.0, 587.0) U/L vs. 259.5 (189.8, 393.8) U/L, P=0.007] were significantly higher in the patients with cardiac involvement. Anti-Ro-52 antibody was detected more commonly in the ASS patients with cardiac involvement compared with the patients without cardiac involvement (91.7% vs. 69.4%, P=0.029). No significant differences were found in the comorbidities, alanine transaminase (ALT), aspartate transaminase (AST), creatine kinase (CK), erythrocyte sedimentation rate (ESR), ferritin (Fer), immunoglobulin G (IgG), complement 3 (C3), complement 4 (C4), lymphocyte subset between the two groups. CONCLUSION: Cardiac involvement is common in ASS, mainly manifested as myocardial damage. It is necessary to be aware of cardiac complications in patients with elevated CRP, elevated LDH and positive anti-Ro-52 antibody.

[Clinical and immunological characteristics of 88 cases of overlap myositis].

OBJECTIVE: To investigate the clinical and immunological characteristics of overlap myositis (OM) patients. METHODS: The data of 368 patients with idiopathic inflammatory myopathies (IIMs) admitted to Peking University People's Hospital from January 2004 to August 2020 were analyzed retrospectively, including demographic characteristics, clinical characteristics (including fever, Gottron' s sign/papules, Heliotrope rash, V-sign, Shawl sign, Mechanic' s hands, skin ulceration, periungual erythema, subcutaneous calcinosis, dysphagia, myalgia, myasthenia, arthritis, Raynaud' s phenomenon, interstitial lung disease, pulmonary hypertension and myocardial involvement), laboratory characteristics, immunological characteristics [including antinuclear antibodies, rheumatoid factors, myositis-associated autoantibodies (MAAs) and myositis-specific autoantibodies (MSAs)] and survival. The clinical and immunological characteristics and prognostic differences of OM and non-OM were compared. The Kaplan-Meier and Log Rank methods were used to analyze the survival. RESULTS: A total of 368 patients were included. 23.9% (88/368) of IIMs patients were OM patients. Among the 88 OM patients, 85.2% (75/88) of them were female, and the median interval between disease onset and diagnosis was 13.5 months. The incidence of overlapped connective tissue diseases in the OM patients was dermatomyositis (DM) in 60.2%, polymyositis (PM) in 3.4%, immune-mediated necrotizing myopathy (IMNM) in 2.3% and anti-synthetase syndrome (ASS) in 34.1%. Compared with the non-OM patients, the proportion of the females in the OM patients was higher (85.2% vs. 72.1%, P=0.016), the OM patients had longer disease duration [13.5(4.5, 48.0) months vs. 4.0(2.0, 12.0) months, P < 0.001]. As for clinical characteristics, compared with the non-OM patients, the incidence of V-sign (25.0% vs. 44.6%, P=0.001) and periungual erythema (8.0% vs. 19.6%, P=0.013) were lower; the incidence of Raynaud's phenomenon (14.8% vs. 1.8%, P < 0.001), interstitial pneumonia (88.6% vs. 72.1%, P=0.001), pulmonary hypertension (22.7% vs. 7.5%, P < 0.001) and myocardial involvement (18.2% vs. 9.3%, P=0.033) were higher. As for immunological characteristics, compared with the non-OM patients, the incidence of elevated aspartate aminotransferase (AST) (31.8% vs. 45.0%, P=0.035) was lower and elevated C-reactive protein (CRP) (58.0% vs. 44.6%, P=0.037) was higher; the positive rates of antinuclear antibodies (ANA) (85.1% vs. 63.4%, P=0.001) and rheumatoid factors (RF) (40.2% vs. 17.8%, P < 0.001) and anti-Ro-52 (71.6% vs. 56.1%, P=0.038) in serum were higher. There was no significant difference in the survival between the OM patients and non-OM patients. CONCLUSION: Pulmonary hypertension and myocardial involvement were frequently observed in OM.

Summer flounder (Paralichthys dentatus) sperm cryopreservation and application in interspecific hybridization with olive flounder (P olivaceus).

The present study established an efficient technology for summer flounder (Paralichthys dentatus) sperm cryopreservation, and successfully applied the cryopreserved sperm into interspecific hybridization with olive flounder (P olivaceus). The best motility of postthaw sperm (78.00 ± 4.70% and 76.60 ± 7.90%), fertilization rates (95.70 ± 3.60% and 79.40 ± 5.20%), and hatching rates (93.10 ± 4.00% and 77.20 ± 2.90%) were achieved when using propylene glycol or DMSO as cryoprotectant. Furthermore, we have successfully improved the cryopreservation method from using 2-mL cryotube to 5-mL cryotube, and the dilution ratio has been increased to 4:1. By this method, the cryopreservation efficiency has been improved to 30 times of that with routine method. Computer-assisted sperm motion analysis showed the freezing-thawing process decreased the sperm speed but did not significantly change the sperm movement pattern, and the progressive linear motion still was the dominant movement pattern. The ultrastructural analysis showed 50% to 60% of the spermatozoa had normal morphology, 20% to 30% were slightly damaged, such as swelling or rupture of head, midpiece, and tail region, and 10% to 20% were severely damaged. In the artificial hybridization experiment, high fertilization rates and hatching rates were achieved when using 15% DMSO (95.7 ± 3.6% and 79.4 ± 5.2%, respectively) and 15% propylene glycol (93.1 ± 4.0% and 77.2 ± 2.9%, respectively), with no significant difference in comparison with control (94.7 ± 2.6% and 72.5 ± 6.5%, respectively). In addition, we found the embryos and larvae from postthaw sperm of P dentatus developed and grew normally. The results of the present study further validated the safety of the cryopreserved sperm in breeding production by assessing the fertilization capacity, embryo development, and larval growth.

Germline-specific and sexually dimorphic expression of a dead end gene homologue in turbot (Scophthalmus maximus).

Germ cells are indispensable for gonadal development and fertility. However, the physiological mechanisms regulating germ cell development in marine fish are poorly understood due to a lack of germ cell markers. The dead end (dnd) gene is a vertebrate-specific component of germplasm crucial for primordial germ cells (PGCs) migration and development in teleosts. In this study, we identified a dnd homologue (Smdnd) in turbot (Scophthalmus maximus) and investigated its expression pattern during embryogenesis and gonadal development. The deduced amino acid sequence of Smdnd shared several conserved motifs of Dnd homologues as well as high identity to other Dnd proteins. Phylogenetic analysis revealed that the SmDnd was closely related to its teleost counterparts. Reverse transcription polymerase chain reaction (RT-PCR) and in situ hybridization revealed that Smdnd transcripts could be exclusively detected in germ cells, including presumptive PGC and adult male and female germ cells. In addition, an interesting sexually dimorphic expression of Smdnd during gonadal development was observed by real-time PCR. Female turbot showed greater (P < 0.05) Smdnd expression than male before sex maturation. This difference reduced gradually due to the upregulation of Smdnd in the male during the period corresponding to spermatogonia proliferation and meiosis. These results indicate that Smdnd can be used as a germ cell marker in turbot. In addition, the temporal and sex differences in Smdnd expression indicate that this gene may play different roles in gonadal development in both sexes.

Germ cells and fertilization differences among Japanese flounder Paralichthys olivaceus, summer flounder Paralichthys dentatus and their first and second generations.

The morphology of gametes and the fertilization biology of Japanese flounder Paralichthys olivaceus (Po), summer flounder Paralichthys dentatus (Pd) and their hybrids were examined. Multiple generations (two hybrids: Poâ™€× Pd♂ (F1) and Pdâ™€× Po♂; two backcrosses: F1â™€× Po♂ and F1â™€× Pd♂) were obtained by artificial insemination. Comparison of egg ultrastructure among Po, Pd and F1 showed the morphology of micropyle region and the distribution density of pores were species specific. There were c. 100-200 accessory openings around the micropyle in Po, but not in Pd and F1. The zona radiata thickness and number of parallel bands were similar between F1 and Po, which were different from Pd. Comparison of spermatozoa ultrastructure revealed a close relationship between Po and Pd. Cytologically, the six crosses obeyed normal fertilization and cleavage processes, and only one male pronucleus was observed in a fertilized egg, indicating a monospermic fertilization pattern. Analysis of the time distribution from fertilization to first cleavage revealed an obvious delay at pronucleus fusion in the Pd × Po cross. The delay might indicate some cytoplasmic-nuclear incompatibility during the process of fertilization.

Skeletal development and abnormalities of the vertebral column and of the fins in hatchery-reared turbot Scophthalmus maximus.

To describe the skeletal development and abnormalities in turbot Scophthalmus maximus, samples were collected every day from hatching to 60 days after hatching (DAH). A whole-mount cartilage and bone-staining technique was used. Vertebral ontogeny started with the formation of anterior haemal arches at 5·1 mm standard length (L(S) ) c. 11 DAH, and was completed by the full attainment of parapophyses at 16·9 mm L(S) c. 31 DAH. Vertebral centra started to develop at 6·3 mm L(S) c. 16 DAH and ossification in all centra was visible at 11·0 mm L(S) c. 25 DAH. The caudal fin appeared at 5·1 mm L(S) c. 11 DAH and ossification was visible at 20·6 mm L(S) c. 37 DAH. The onset of dorsal and anal fin elements appeared at 5·8 mm L(S) c. 15 DAH and 6·3 mm L(S) c. 16 DAH, respectively. Ossifications of both dorsal fin and anal fin were visible at 20·6 mm L(S) c. 37 DAH. The pectorals were the only fins present before first feeding, their ossifications were completed at 23·5 mm L(S) c. 48 DAH. Pelvic fins began forming at 7·2 mm L(S) c. 19 DAH and calcification of the whole structure was visible at 19·8 mm L(S) c. 36 DAH. In the present study, 24 types of skeletal abnormalities were observed. About 51% of individuals presented skeletal abnormalities, and the highest occurrence was found in the haemal region of the vertebral column. As for each developmental stage, the most common abnormalities were in the dorsal fin during early metamorphic period (stage 2), vertebral fusion during climax metamorphosis (stage 3) and caudal fin abnormality during both late-metamorphic period (stage 4) and post-metamorphic period (stage 5). Such research will be useful for early detection of skeletal malformations during different growth periods of reared S. maximus.

Study on permeability of DMSO in embryos of red seabream (Pagrus major) by capillary electrophoresis.

The objectives were to investigate the permeability of DMSO to red seabream (Pagrus major) embryos by capillary electrophoresis and the effects of DMSO concentrations (5 to 40 percent, volume basis) and immersion times (10, 30 and 60 min) on hatching rate and morphology. The results suggested the internal DMSO concentrations were positively related with the external concentrations and exposure times, while the hatching rate was negatively related. The hatching rate decreased drastically (less than 50 percent) after exposure in 35 percent, 20 percent and 15 percent DMSO for over 10, 30 and 60 min, respectively. In all groups, when hatching rate was greater 50 percent, the internal DMSO concentration was less than 2 percent, which was still insufficient for successful cryopreservation. Morphological changes indicated the chorion was permeable to the cryoprotectant. A sign of dehydration in yolk were observed, for a significant decrease in the maximal yolk sac diameter. However, further research was needed to investigate whether the DMSO permeated into the yolk.