Glutamate-ammonia ligase (GLUL, also known as glutamine synthetase) is a crucial enzyme that catalyzes ammonium and glutamate into glutamine in the ATP-dependent condensation. Although GLUL plays a critical role in multiple cancers, the expression and function of GLUL in gastric cancer remain unclear. In the present study, we have found that the expression level of GLUL was significantly lower in gastric cancer tissues compared with adjacent normal tissues, and correlated with N stage and TNM stage, and low GLUL expression predicted poor survival for gastric cancer patients. Knockdown of GLUL promoted the growth, migration, invasion and metastasis of gastric cancer cells in vitro and in vivo, and vice versa, which was independent of its enzyme activity. Mechanistically, GLUL competed with β-Catenin to bind to N-Cadherin, increased the stability of N-Cadherin and decreased the stability of β-Catenin by alerting their ubiquitination. Furthermore, there were lower N-Cadherin and higher β-Catenin expression levels in gastric cancer tissues compared with adjacent normal tissues. GLUL protein expression was correlated with that of N-Cadherin, and could be the independent prognostic factor in gastric cancer. Our findings reveal that GLUL stabilizes N-Cadherin by antagonizing β-Catenin to inhibit the progress of gastric cancer.

Retzius-sparing robot assisted radical prostatectomy (RS-RARP) can significantly improve the immediate urinary continence without increasing the positive rate of surgical margin.However, the learning curve is long, and fewer than 10% of the surgeons can master it.Therefore,we have optimized the procedures of RS-RARP, applying radical prostatectomy with retrograde release of neurovascular bundle to preserve it to the maximum extent.Urethral anastomosis can be performed with only one suture, which eliminates the need for Hem-o-lok and reduces subsequent complications.Our team routinely carries out this operation, and conlcudes that this surgical method can achieve good tumor control, good urinary continence, fast recovery of sexual function, few complications, and strong operability.This article details the key steps and operation experience of this technique.

Objective:This study aims to evaluate the performance of digital PCR (dPCR) detecting multiple and single copies genes of the Epstein-Barr virus (EBV) for nucleic acid quantification and explore their applicability in clinical settings.Methods:Compared the sensitivity, specificity, precision, lower limit of detection (LoD), and linearity for multicopy BamHI-W dPCR and single-copy EBNA1 dPCR systems. Linear regression analysis using the least squares method was employed to evaluate the linearity. Additionally, we analyzed plasma samples from 182 patients with suspected EBV-related diseases between January and July 2022 at the Southern Medical University Southern Hospital, using both dPCR and quantitative PCR (qPCR) for EBV DNA quantification. Linear regression analysis using the least squares method was conducted to assess their quantitative correlation.Results:The dPCR systems for both multicopy and single-copy genes showed excellent linearity ( R 2 values of 0.992 and 0.997, respectively, both P<0.001). The LoD were 188 IU/ml for BamHI-W gene and 358 IU/ml for EBNA1 gene dPCR systems. The logarithmic coefficient of variation ( CV) values for high-concentration samples (1 000 000 IU/ml) were 0.34% and 0.21% for the BamHI-W gene and EBNA1 gene dPCR assays, respectively, while for low-concentration samples (5 000 IU/ml) were 0.98% and 0.64%, respectively. In the detection of seven common clinical infectious pathogens and EBV positive samples, only EBV-positive samples yielded positive signals in the dPCR detection system, with no cross-reaction with other pathogens. In 182 samples, the positive detection rates were 47.80% (87/182) for BamHI-W gene and 35.16% (64/182) for EBNA1 gene dPCR, compared to 43.41% (79/182) for qPCR. Linear correlation analysis with qPCR showed R2 values of 0.837 for BamHI-W gene and 0.763 for EBNA1 gene dPCR (both P<0.001). The BamHI-W gene copy number ranged from 3 to 18 copies per clinical sample, with patient-specific variations. There was a high consistency in viral load trends between the multicopy BamHI-W gene and single-copy EBNA1 gene dPCR systems within individual patients. Conclusions:The dPCR methods detecting EBV multiple and single copies genes showed high sensitivity, specificity, precision, and quantitative accuracy, suitable for clinical sample analysis. The multicopy BamHI-W gene dPCR method notably enhances detection sensitivity and can be used as a supplement to current EBV DNA load detection methods, especially in low-concentration samples. For within-patient EBV DNA monitoring, the multicopy gene method proves more effective, while inter-patient comparisons might necessitate single-copy gene methods or normalize them using the same standard.

Objective:To explore the clinical efficacy of retzius-sparing robot-assisted laparoscopic radical prostatectomy with retrograde release of the neurovascular bundle.Methods:From January 2021 to January 2022, the clinical data of 113 patients with retzius-sparing robot-assisted laparoscopic radical prostatectomy (RS-RARP) was retrospectively analyzed. The ages of the optimized group and the standard group were (67.5±6.4) years and (67.7±6.1) years, and the body mass index (BMI) was (25.0±3.2) kg/m 2 and (24.9±3.1) kg/m 2, respectively. The prostate volume was (42.8±15.4) ml and (41.0±17.9) ml, the preoperative PSA was (13.5±13.1) ng/ml and (11.9±16.0) ng/ml, and the preoperative IIEF-5 score was (15.0±4.0) and (14.8±4.2) points, respectively. Gleason scores were (7.2±0.8) points and (7.1±0.9) points, respectively, with no statistical significance ( P>0.05). The clinical stages of cT 2 and cT 3 were 35 and 40 cases in the optimized group and 16 and 22 cases in the standard group, respectively. There were 5 and 11 cases of preoperative neoadjuvant therapy, respectively, with no statistical significance ( P>0.05). The optimized RS-RARP is the blunt dissection of the denonvilliers fascia and forward to the apex of prostate, and retrograde release of the NVB. The operation time, intraoperative blood loss, drainage tube retention time, days of hospital stay, positive rate of pathological margin and incidence of complications were compared. The recovery of urinary continence was evaluated at 2 weeks after the catheter was removed, and the postoperative IIEF-5 score and PSA were followed up at 1 month after the surgery. Immediate urinary continence was defined as the use of 1 pad on the day of radical prostatectomy. Follow-up intervals were no more than 3 months. The log-rank test compared urinary incontinence rates between the two modalities. Results:All 113 cases of surgery were completed, and median follow-up was 16 months. The operation time was (79.7±26.6) min and (149.8±40.1) min, and the intraoperative blood loss was (54.9±24.7) ml and (110.0±83.2) ml, respectively, and the difference was statistically significant ( P<0.01). The retention time of postoperative drainage tube was (5.3±2.1) d and (5.5±2.1) d in the optimal group and the standard group, and the days of hospital stay was (7.6±2.1) d and (8.5±2.3) d, respectively. The positive rate of postoperative pathological margin was 19.6% (10/51) and 24.2% (15/62), respectively. There was no significant difference ( P>0.05). immediate urinary continence was 86.3% (44/51) and 69.4% (43/62) in the optimized group and the standard group, respectively, and the difference was statistically significant ( P=0.033). Postoperative IIEF-5 scores were (13.2±3.3) and (11.0±4.3), respectively, and the difference was statistically significant ( P=0.012). Kaplan-Meier analysis showed that the risk of urinary incontinence was lower in the optimized group ( P=0.02). Conclusions:The optimized RS-RARP might shorten the operation time, reduce intraoperative bleeding, and help the recovery of urinary continence and sexual function to a great extent.

Objective:To summarize and analyze the clinicopathological characteristics of patients with DNAJ heat shock protein family member B9 (DNAJB9)-positive fibrillary glomerulonephritis (FGN).Methods:The clinical and pathological data of 5 patients with DNAJB9-positive FGN diagnosed in Peking University First Hospital from January 2011 to January 2021 were retrospectively collected and analyzed.Results:Among the 5 patients, the female to male ratio was 4∶1, and the median age was 29 years old (24-71 years old). The clinical manifestations included 2 cases with nephrotic syndrome and 3 cases with proteinuria. One patient had gross hematuria, and 4 cases had mild microscopic hematuria. None of the 5 patients had evidence of monoclonal gammopathy. The renal pathological pattern of FGN showed mesangial-proliferative glomerulonephritis, mesangial nodular sclerosis, membranoproliferative glomerulonephritis, and atypical membranous nephropathy. Crescents formation could be accompanied. Immunofluorescence staining showed smudgy and granular IgG and C3 deposition in the mesangial region and capillary wall, and the subtypes of IgG were mainly IgG1 and IgG4. Under electron microscopy, fibrillary deposits with a diameter of 8-30 nm were observed in the mesangial and subendothelial area, accompanied by deposition in basement membrane and occasionally subepithelial area. The renal prognosis of FGN patients was poor. One patient entered end-stage renal disease within one week, and another patient entered end-stage renal disease within one year despite immunosuppressant therapy in 2 cases with nephrotic syndrome at onset. One patient had worsening proteinuria despite renin-angiotensin system (RAS) blocker treatment. Two patients achieved complete renal remission and stable renal function after RAS blocker treatment.Conclusions:Most FGN patients in China are young people. The main clinical manifestations are proteinuria or mild microscopic hematuria. The diagnosis depends on the discovery of fibrillary deposits in the mesangial area and subendothelial area with a diameter of about 10-30 nm under the electron microscope. DNAJB9 protein immunohistochemical staining can be used as an important marker for the diagnosis of FGN. The prognosis of FGN kidney is poor, and there is no effective targeted treatment option now.

Objective:To investigate the efficacy and prognosis of hypofractionated intensity-modulated radiation therapy combined with hormonal therapy in the treatment of pelvic lymph node metastatic prostate cancer.Methods:Clinical data of 42 IV A prostate cancer patients who received hypofractionated intensity-modulated radiation therapy combined with hormonal therapy in Cancer Hospital of Chinese Academy of Medical Sciences between 2006 and 2018 were retrospectively analyzed. The total irradiation doses to the prostate and seminal vesicles were 67.5 Gy/25f, 2.7 Gy/f. The prophylactic irradiation doses to the pelvic lymph nodes were 45-50 Gy with a daily fraction dose of 1.8-2.0 Gy. Thirty-three patients with residual lymph nodes were boosted to 60.0-67.5 Gy for the residual area, 2.4-2.7 Gy/f. Androgen deprivation therapy included surgical castration or luteinizing hormone-releasing hormone agonists combined with antiandrogens. Survival rate was calculated using Kaplan- Meier method. The differences between two groups were analyzed by log-rank test. Prognostic factors were identified by univariate and multivariate analyses. Results:The median follow-up was 65.5 months (range, 5 to 150 months). The 5-year and 10-year failure-free survival (FFS) rates in the whole group were 67% and 45%, respectively. No clinical recurrence was observed in the irradiation field. The 5-year and 10-year prostate cancer-specific survival/overall survival (PCSS/OS) rates were 85% and 60%, respectively. Gleason score (≥8 and<8) and duration of hormonal therapy impacted the FFS (both P<0.05). The duration of hormonal therapy was an independent prognostic factor for PCSS/OS ( P=0.003). Conclusions:Hypofractionated intensity-modulated radiotherapy combined with hormonal therapy yields optimistic clinical efficacy in the treatment of pelvic lymph node metastatic prostate cancer. Gleason score (≥8 and <8) and duration of hormonal therapy are critical prognostic factors.

Objective:The clinical heterogeneity of non-metastatic castration-resistant prostate cancer is high, and precise and individualized treatment is required for different patients to achieve maximum benefits. Three cases of non-metastatic castration-resistant prostate cancer were reported in this paper. One case received apalutamide + leproprillin treatment, one received radical prostatectomy, and one received radiotherapy + abiraterone treatment. After a period of follow-up, the three patients all benefited to varying degrees.

Objective:To evaluate the toxicities and clinical efficacy of postoperative radiotherapy for children with nephroblastoma (Wilms’ tumor).Methods:In total, 116 WT Children (≤14-year-old) treated with radiotherapy (RT) in our center from 2005 to 2018 were recruited in this retrospective analysis. RT-induced toxicities and clinical efficacy were analyzed. RT was performed guided by Children′s Oncology Group (COG) protocol. The overall survival (OS), flank-field control (LC), abdominal control (AC), and distant metastasis-free survival (DMFS) were calculated using the Kaplan-Meier method. pathologically proved.Results:From January, 2005 to August, 2018, 116 pathologically proved WT patients were enrolled. Most of them were diagnosed with favorable WT (94.8%) and stage Ⅲ WT (87.1%). With a median follow-up time of 30.4(0.7-185.7) months, the 3-year OS, LC, AC and DMFS were 83.9%, 78.2%, 75.2% and 82.8%, respectively. Sixty-four (55.2%) patients suffered from Grade I to Ⅱ gastrointestinal toxicities and Grade I to IV hematological toxicities. Only 5 patients (4.3%) had Grade Ⅱ late toxicities. For 96 patients who received adjuvant RT, the median surgery-RT interval time was 1.2(0.5 to 7.1) months. The 3-year OS, LC, AC and DMFS were 88.1%, 96.7%, 92.7% and 86.9%, respectively. Patients with tumor rupture without whole abdomen irradiation (WAI) tended to have lower AC, DMFS and OS. Twenty children received salvage RT when they had disease relapse. The 3-year OS and DMFS of patients with salvage RT were significantly worse than those receiving adjuvant RT (OS: 68.2% vs. 88%, P=0.012; DMFS: 64.3% vs. 86.9%, P=0.032). Conclusions:Tumor bed irradiation for WT patients as per COG protocol can be well tolerated and achieve high efficacy. Salvage RT yields poor efficacy for tumor bed recurrence. Furthermore, tumor rupture without WAI possibly increases the abdominal and distant recurrence and the risk of death.