Follow-up analysis of 40 cases of robotic intracorporeal studer orthotopic neobladder in a single center / 中华泌尿外科杂志

Objective:To explore the curative effect of robotic intracorporeal studer orthotopic neobladder (RISON).Methods:The clinical data of 40 patients who underwent RISON in our hospital from April 2018 to March 2020 were analyzed retrospectively, including 39 males and 1 female with an average age of (56.4±9.9)years, an average body mass index of (25.5±3.1) kg/m 2. Four patients received preoperative neoadjuvant chemotherapy. Comorbidity component index score was 0-2 points in 6 cases, 3-5 points in 33 cases, 6-8 points in 1 case. All patients had definite pathological diagnosis for the high grade urothelial carcinoma or recurrent bladder tumors, and the tumor staging forecast within T 2 stage. Biopsy of the posterior urethra and bladder neck revealed negative result of tumor invasion. The patients had normal kidney function and without any clear bowel disease history. All patients underwent robotic radical intracorporeal studer orthotopic neobladders and standard lymphadenectomy. About 30 to 40 cm from terminal ileum was selected as the new bladder neck and 50 cm ileum as neobladder. Restore intestinal continuity with EndoGIA. The intestinal canal was cut off along the offside of mesentery except for proximal 10 cm. After U-shape suture of the new bladder back wall, the new anterior bladder wall was sutured with barbs line further in pulp muscularis. "Wallace" was performed bilateral ureteral anastomosis and closing the proximal end of the anterior wall of the new bladder. Further follow-up results were also collected to evaluate the clinical treatment effect, including pathological results of tumor, bladder capacity, urine control (usage of pads), sexual function, short-term(≤30 d) and long-term(>30 d) complications. Results:All the RISONs were successfully performed without conversion to open operation. Median operation time was 360(300.0, 442.5) min, blood loss was 200(200.0, 337.5) ml, indwelling time of gastric tube was 3(3, 4)d, full tolerance time was 3(3, 5)d, and postoperative hospital stay was 9(8, 10)d. Postoperative pathological results showed 39(97.5%) cases were no more than T 2N 0M 0, 1 case was confirmed T 3aN 0M 0 pathologically, 1 case was positive in surgical margin, 1 case was diagnosed as incidental prostate cancer, 15(12, 20) lymph nodes were dissected and no lymph node metastasis was found. Short-term (within 30 days) complications occurred in 12 patients, including Clavien grade Ⅰ 7 cases, Clavien grade Ⅱ 5 cases. Long-term (out of 30 days) complications occurred in 18 patients, including Clavien grade Ⅰ 10 cases, Clavien grade Ⅱ 7 cases, Clavien grade Ⅲ 1 case. And the follow-up time was 1 to 24 months. The median bladder volume 300 (0, 400)ml of 25 patients who were followed up for more than one year and 1 in 34 female patient developed incontinence. Other 33 patients had continence rate of 93.9% (0 pad) during the day, and needed regular urination at night (1-3 times), with an average of 2 pads. All 11 patients with nerve-sparing achieved satisfactory erectile function after operation, with an average IIEF-6 score of (21.5±2.7). No tumor recurrence or death occurred in all patients during the follow-up period. Conclusions:Robotic intracorporeal studer orthotopic neobladder is a safe and feasible urinary diversion operation. The patients achieved good clinical efficacy in tumor control, bladder volume, daytime and nighttime continence, sexual function recovery.

Agrobacterium-mediated transformation of LJAMP2 gene into 'Red Sun' kiwifruit and its molecular identification / 生物工程学报

Bacterial canker caused by Pseudomonas syringae pv. Actinidiae is one of the most important diseases of kiwifruit (Actinidia chinensis) and leads to considerable yield losses. In order to obtain transgenic plants with resistance for 'Red Sun' kiwifruit to canker disease, a non-specific lipid transfer protein-like antimicrobial protein gene (LJAMP2) from motherwort (Leonurus japonicus) was introduced into 'Red Sun' kiwifruit through Agrobacterium-mediated transformation. After two days of co-cultivation with A. tumefaciens strain LBA4404 harboring 35S:LJAMP2, the transformed explants were transferred to the selection medium containing 25 mg/L kanamycin+3.0 mg/L BA+1.0 mg/L NAA. The regeneration efficiency of kanamycin-resistant shoots reached to 85%. All (100%) of kanamycin-resistant shoots rooted on half-strength MS medium supplemented with 0.8 mg/L IBA and a total of 40 regenerated plantlets were obtained. PCR and histochemical GUS activity analysis show that 23 of 40 lines (57.50%) were positive, suggesting that the LJAMP2 gene was integrated into the genome of 'Red Sun' kiwifruit. Taken together, we established an efficient genetic transformation method for 'Red Sun' kiwifruit using A. tumefaciens and the transformation frequency reached 5.11%. This protocol will be useful for the genetic breeding of 'Red Sun' kiwifruit for improvement of disease resistance.

Establishment of high frequency regeneration via leaf explants of 'Red Sun' kiwifruit (Actinidia chinensis) / 生物工程学报

A high efficient in vitro regeneration protocol was developed from leaf explants of the female 'Red Sun' kiwifruit (Actinidia chinensis) and the multiplication coefficient and rooting rate of adventitious buds were also optimized. This method does not require formation of callus tissues which leads to somaclonal variations. The results show that the adventitious buds developing directly from explants tissue were noticed after 30 d of culture. The maximum regeneration frequency of adventitious buds is 100% and 18.67 shoots was observed in each leaf explants when MS medium was supplemented with 3.0 mg/L BA+1.0 mg/L NAA. The optimal culture medium for bud multiplication is MS+2.0 mg/L BA+1.0 mg/L NAA+0.1 mg/L GA3 and the multiplication coefficient reached 8.63. On the rooting medium with 1/2 MS+0.8 mg/L IBA for 15 d, the adventitious plantlets were transferred into matrix perlite supplied with 1/2 MS liquid medium for 15 d and the rooting rate reached 100%. 95 out of 98 plantlets (96.94%) survived acclimatization, producing healthy plants in the greenhouse. Taken together, a highly efficient regeneration method via leaf explants of 'Red Sun' kiwifruit was successfully established. This protocol may be useful for micropropagation and genetic transformation studies of 'Red Sun' kiwifruit.