Microscopic Replantation of Complete Penile Amputation With Video Demonstration.

BACKGROUND: Penile amputation is an extremely rare genital injury. To the best of our knowledge, there are only about 200 cases reported in Chinese and English literature, most of them are case reports. So far, there is not any video demonstration of microscopic replantation of complete penile amputation with meticulous surgical skills. OBJECTIVE: To provide a successful example of penile replantation after complete penile amputation through video presentation of the application of meticulous microsurgical techniques and optimized procedures. MATERIALS AND METHODS: The 25-year-old patient was admitted to our hospital 3.5 hours after his penis was completely amputated due to self-mutilation. Microscopic penile replantation was immediately performed after preoperative preparation. After the surgical procedure, the patient was treated with broad-spectrum antibiotics, analgesia, antithrombotics and anxiolytic. RESULTS: The total ischemic time was about 10 hours. The duration of surgery was about 7 hours. On the 14th day post-surgery, the wound healed smoothly, the glans was ruddy in color, and the appearance returned to normal without obvious complications. The patient urinates normally with a maximal urinary flow rate of 25 ml/s after removing the catheter. Three months after surgery, the local sensation of foreskin and glans recovered significantly, which showed that slight needling could lead to obvious pain, and the penis erection hardness score was 3 during morning erection or urinary bladder distention. Six months after surgery, the patient reported that he was completely satisfied with the result, which showed that the sensation of the penis and glans surface returned to almost normal and the optimal erection hardness score was 4. CONCLUSION: Careful microsurgical anastomosis of the dorsal arteries, deep dorsal vein, superficial dorsal vein and multiple dorsal nerves could obtain ideal recovery of penile appearance and function and avoid any obvious complications.

Neoadjuvant Chemotherapy Improves the Immunosuppressive Microenvironment of Bladder Cancer and Increases the Sensitivity to Immune Checkpoint Blockade.

Although tumor immune microenvironment plays an important role in antitumor therapy, few studies explored the gene signatures associated with the tumor immune microenvironment of bladder cancer after neoadjuvant chemotherapy. We examined and analyzed differentially expressed genes from 9 patients with stage I-III bladder cancer by RNA immune-oncology profiling platform. After neoadjuvant chemotherapy, the expressions of 43 genes in 19 pathways and 10 genes in 5 pathways were upregulated and downregulated, respectively. Neoadjuvant chemotherapy also promoted the expression of genes related to the activation of antitumor immune responses and decreased the expression of genes related to tumor proliferation pathways. In addition, neoadjuvant chemotherapy improved tumor response to immune checkpoint blockade. Furthermore, this study also identified several genes that can be used to predict the efficacy of neoadjuvant chemotherapy and their possible molecular mechanisms. In conclusion, neoadjuvant chemotherapy may promote the activation of antitumor effects, improve the suppressive tumor immune microenvironment, and increase the sensitivity of bladder cancer to immune checkpoint blockade.

Upregulation of PD-L1 and APE1 is associated with tumorigenesis and poor prognosis of gastric cancer.

INTRODUCTION: Gastric cancer is a fatal malignancy with a rising incidence rate. Effective methods for early diagnosis, monitoring metastasis, and prognosis are currently unavailable for gastric cancer. In this study, we examined the association of programmed death ligand-1 (PD-L1) and apurinic/apyrimidinic endonuclease 1 (APE1) expression with the prognosis of gastric cancer. METHODS: The expressions of PD-L1 and APE1 were detected by immunohistochemistry in 107 cases of human gastric carcinoma. The correlation of PD-L1 and APE1 expression with the clinicopathologic features of gastric carcinoma was analyzed by SPSS version 19.0. RESULTS: The positive expression rates of PD-L1 and APE1 in gastric cancer tissues were 50.5% (54/107) and 86.9% (93/107), respectively. PD-L1 and APE1 positive expressions were significantly associated with depth of invasion, lymph node metastasis, pathological type, overall survival, and higher T stage. Furthermore, the expression of PD-L1 in highly differentiated gastric cancers was higher than that in poorly differentiated cancers (P=0.008). Moreover, the expression of APE1 and PD-L1 in gastric cancers was positively correlated (r=0.336, P<0.01). Multivariate analysis showed that the depth of invasion was a significant prognostic factor (risk ratio 19.91; P=0.000), but there was no significant relationship with PD-L1, APE1, prognosis, and other characteristics. CONCLUSION: The deregulation of PD-L1 and APE1 might contribute to the development and the poor prognosis of gastric cancer. Our findings suggest that high expression of PD-L1 and APE1 is a risk factor of gastric cancer and a new biomarker to predict the prognosis of gastric cancer. Furthermore, our findings suggest that targeting the PD-L1 and APE1 signaling pathways may be a new strategy for cancer immune therapy and targeted therapy for gastric cancer, especially in patients with deep invasion and lymph node metastasis.

Cloning and functional expression of a human lysozyme gene (hly) from human leukocytes in Pichia pastoris.

hly is a cDNA gene derived from human leukocytes that encodes a mature human lysozyme (abbreviated to hLY). The aim of the present study was to determine the effect of cloned hly on recombinant hLY (r-hLY) activity under optimized conditions. hly was amplified by RT-PCR and ligated into the pPIC9K plasmid. The cloned cDNA (hly) was 393 bp in length, encoding a 130 amino acid hLY with a calculated molecular mass of 14,698 Da. The recombinant expression plasmid, designated as pPIC9K-hly, was linearized with SacI and transformed into Pichia pastoris GS115 (his4, Mut+) by electroporation. The integration of hly into the P. pastoris genome was confirmed by PCR analysis using 5'-AOX1 and 3'-AOX1 primers. Yeast extract peptone dextrose (YPD) plates containing different concentrations of geneticin (G418) were used for the screening of P. pastoris transformants (His+, Mut+) with multiple hly copies. One transformant resistant to 4.0 mg/ml of G418, designated as P. pastoris GShLY4-6, expressing the highest r-hLY activity was selected by the shake-flask test, and used for the optimization of expression conditions. When the P. pastoris GShLY4-6 was induced under optimized conditions, the expressed r-hLY activity was up to 533 U/ml, which was 1.52 times as high as that (351 U/ml) expressed using the standard protocol. SDS-PAGE assay demonstrated that the r-hLY with an apparent molecular mass of approximately 14.7 kDa was extracellularly expressed in P. pastoris. In conclusion, r-hLY increased following the cloning of hly and the optimized conditions as compared to standard protocol.