Coronaviruses using different strategies to antagonize antiviral responses and pyroptosis

Viral infection triggers inflammasome-mediated caspase-1 activation. However, less is known about how viruses use the active caspase-1 to evade host immune response. Here, we use porcine epidemic diarrhea virus (PEDV) as a model of coronaviruses (CoVs) to illustrate the sophisticated regulation of CoVs to counteract IFN-I signaling and pyroptosis. We show that PEDV infection stabilizes caspase-1 expression via papain-like protease PLP2s deubiquitinase activity and the enhanced stabilization of caspase-1 disrupts IFN-I signaling by cleaving RIG-I at D189 residue. Meanwhile, PLP2 can degrade GSDMD-p30 by removing its K27-linked ubiquitin chain at K275 to restrain pyroptosis. Papain-like proteases from other genera of CoVs (PDCoV and SARS-CoV-2) have the similar activity to degrade GSDMD-p30. We further demonstrate that SARS-CoV-2 N protein induced NLRP3 inflammasome activation also uses the active caspase-1 to counter IFN-I signaling by cleaving RIG-I. Therefore, our work unravels a novel antagonistic mechanism employed by CoVs to evade host antiviral response.

The nonstructural protein 5 of coronaviruses antagonizes GSDMD-mediated pyroptosis by cleaving and inactivating its pore-forming p30 fragment

Coronaviruses (CoV) are a family of RNA viruses that typically cause respiratory, enteric and hepatic diseases in animals and humans. Here, we used porcine epidemic diarrhea virus (PEDV) as a model of coronaviruses (CoVs) to illustrate the reciprocal regulation between CoVs infection and pyroptosis. For the first time, we clarified the molecular mechanism of porcine Gasdermin D (pGSDMD)-mediated pyroptosis and demonstrated that amino acids T239 and F240 within pGSDMD-p30 are critical for pyroptosis. Furthermore, 3C-like protease Nsp5 from SARS-CoV-2, MERS-CoV, PDCoV and PEDV can cleave human/porcine GSDMD at the Q193-G194 junction upstream of the caspase-1 cleavage site to produce two fragments which fail to trigger pyroptosis or inhibit viral replication. Thus, we provide clear evidence that coronoviruses may utilize viral Nsp5-GSDMD pathway to help their host cells escaping from pyroptosis, protecting the replication of the virus during the initial period, which suggest an important strategy for coronoviruses infection and sustain.

High power lateral green laser assisted non-blocking laparoscopic partial nephrectomy for T 1a renal tumors / 中华泌尿外科杂志

Objective:To explore the safety and efficacy of laparoscopic non-blocking partial nephrectomy assisted by high power lateral green laser in the treatment of T 1a renal tumor. Methods:The clinical data of 10 patients with T1a stage renal tumor from February 2021 to April 2021 in department of urology, Gongli hospital affiliated to Naval Military Medical University were retrospectively analyzed. There were 7 males and 3 females, aged 47.0-74.0 years, with average of(58.8±9.7)years old. The diameter of the tumor ranged from 2.0 cm to 3.8 cm, with an average of (3.1±0.6)cm. There were 6 cases on the left side and 4 cases on the right side, locate on lumbar side in 9 cases and ventral sied in 1 case. The R. E.N.A.L score was 4.0-6.0, with an average of (5.0±0.8). The preoperative creatinine was 66.9-90.1μmol/L, with an average of (75.1±9.0)μmol/L, preoperative GFR of 44. 6- 67. 3 ml /min, with an average of(56.7±7.7)ml/min, preoperative hemoglobin level of 119.0-156.0g/L, with an average of (135.8±11.4)g/L. All patients underwent laparoscopic non-blocking partial nephrectomy assisted by 180w lateral green laser, free the surrounding area of the tumor fully and completely expose the renal tumor. The laser fiber was placed through the green laser hand piece, and the fiber was connected with normal saline to wash the strip. The initial green laser vaporization power was set at 80W, and the hemostasis power at 35W.About 3mm away from the edge of the tumor, and one optical fiber away from the renal parenchyma, the renal parenchyma was cut with 80W power. In order to reduce the interference by smoke, high-pressure flushing was used through the optical fiber while vaporizing, and an attractor was used to push and peel the tumor. In case of bleeding during operation, hemostatic power can be used to close the bleeding point and gradually advance until the tumor was completely removed. The wounds of renal inner medulla and renal outer cortex were continuously sutured in 1-3 layers with barbed suture. It involved 9 cases via retroperitoneal approach and 1 case via abdominal approach. The operation time, postoperative hemoglobin decrease, extraction time of negative pressure drainage, postoperative hospital stay, postoperative pathology and postoperative complications were recorded, and the serum creatinine level and GFR level of the affected side were followed up 1 month after operation.Results:All the operations were successfully completed, and there was no conversion to open surgery or radical nephrectomy. One case changed to scissors fast resection and sutured hemostasis due to severe intraoperative bleeding. The operation time was 90.0-120.0 min, with the average of (104.5±9.0)min. The postoperative hemoglobin level was 96.0-132.0g/L, with an average of (115.2±11.8)g/L, and the difference was statistically significant ( P<0.05). The postoperative hemoglobin decreased from 12.0g/L to 25.0g/L, with an average of (20.6±4.6)g/L. The time of vacuum drainage was 5.0-7.0 days, with an average of (5.7±0.7)d. Postoperative hospital stay was 6.0-8.0 days, with an average of (6.7±0.7)d. No bleeding, urinary leakage and other complications occurred in all patients. There were 7 cases of clear cell carcinoma, 2 cases of papillary renal cell carcinoma and 1 case of angiomyolipoma. All margins were negative. One month after operation, creatinine ranged from 66.0 to 90.4μmol/L, with an average of (76.8±8.3)μmol/L, which was not significantly different compared with that before operation ( P>0.05). One month after operation, GFR was 45.1-60.8 ml/min, and with an average of (55.5±4.7)ml/min, and there was no significant difference compared with preoperative data( P>0.05). Conclusions:For T 1aN 0M 0 stage and exophytic renal tumors, laparoscopic non-blocking partial nephrectomy assisted by lateral green laser is safe and effective.

Initial application experience of six-step method high power side-emitting greenlight laser transurethral anatomical vapor incision technique for the treatment of benign prostate hyperplasia / 中华泌尿外科杂志

Objective:To explore the efficacy and safety of transurethral anatomical vapor incision technique of prostate (VIT) with six-step method high power side-emitting greenlight laser in the treatment of benign prostatic hyperplasia (BPH).Methods:A retrospective analysis of 82 patients with BPH who used high power side-out green laser in the treatment from October 2018 to June 2020 in Gongli Hospital of Naval Medical University was performed. Among them, 40 patients were treated with six-step method VIT, and 42 patients were treated with photoselective vaporization of prostate (PVP). The two groups of patients were compared in age [(71.1±8.7)years vs.(72.1±7.0)years], prostate volume [75 (68.25, 89.00) ml vs. 73 (63.25, 85.00) ml], and peak urinary flow rate (Q max) [6.20 (5.20, 8.20) ) ml/s vs. 5.9 (4.75, 7.50) ml/s], post-void residual volume (PVR) [74.00 (42.50, 103.75) ml vs. 67.00 (58.00, 84.50) ml], international prostate symptom score (IPSS) [(21.2±5.2) vs. ( 21.0±3.9)], quality of life score (QOL) [5 (4, 6) vs. 5 (4, 6) ], prostate specific antigen (PSA) [6.20 (4.12, 8.43) ng/ml vs. 5.40 (3.88, 7.13) ng/ml ]. In general, there was no statistical difference ( P>0.05). The VIT group adopts the six-step method of marking, removing film, grooving, excision, trimming and crushing. In the PVP group, the prostate tissue was uniformly vaporized layer by layer from the inside to the outside. Perioperative indexes and complications were compared between the two groups. The Q max, IPSS, QOL, PVR and PSA between the two groups before and 3 months after surgery were compared. Results:All patients in the VIT group and PVP group successfully completed the surgery, and there was no case of transfer to TURP or open surgery. The average operation time was [60.00(50.00, 73.75)min vs. 70.00(50.00, 73.75)min] ( P<0.05). There was no significant difference in the amount of postoperative hemoglobin decline[15.00(10.00, 17.75)g/L vs. 16.00(14.00, 19.25)g/L], average bladder irrigation time[1(1, 1)d vs. 1(1, 1)d], indwelling catheterization time[3(3, 3)]d vs. 3(3, 3)d] and hospitalization time in patients after operation[4(3, 4)d vs. 4(4, 4)d] ( P>0.05). All patients had no blood transfusion, second bleeding, readmission, TURS, urethral stricture and urinary incontinence.There were 2 cases (5.0%) of postoperative urinary tract infection in the VIT group and 9 cases (21.4%) of postoperative urinary tract infection in the PVP group ( P<0.05), and they were cured after anti-inflammatory treatment. Three months after operation, Q max, IPSS, QOL, PVR and PSA in the two groups were significantly improved compared with preoperatively. Among them, the differences of IPSS [(5.7±2.5) points vs. (7.5±2.8) points] and PSA [2.65(2.10, 3.90)ng/ml vs. 4.00(2.45, 4.45)ng/ml] in the VIT group and PVP group after operation were statistically significant ( P<0.05). Conclusions:Applying the six-step method high power side-emitting greenlight laser transurethral anatomical VIT to treat BPH, there is less intraoperative and postoperative bleeding, short operation time, significant decrease in PSA, and fewer complications. It is a safe and effective minimally invasive technology for the treatment of BPH.

Comparison the efficacy and safety of lithotriptors with different focus size and energy in the management of renal calculi / 实用医学杂志

Objective To compare the clinical efficacy and safety of wide-focus high energy lithotriptor with narrow-focus low energy lithotriptor treating renal calculi. Methods A prospective study was conducted to compare both modalities for the management of renal calculi.Stone formers were randomly enrolled into two groups. Group A was managed with wide-focus high energy lithotriptor,while group B was managed with the other. Urine samples were collected to detect neutrophil gelatinase-associated lipocalin(NGAL)and α1-microglobulin(α1-MG)levels before and after SWL. Results 60 were randomized to the group A and 60 to group B. There was no significant difference between two groups in stone free rate and complication rate.For stones more than 10 mm,re-treat rate(11.1%vs 39.1%,P=0.021)was lower and complication rate(25.9%vs 4.3%,P=0.038)was higher in group A. There had a larger increase of NGAL in group A(P < 0.001)after SWL. Conclusion SWL with both lithotriptors are effective and safe for renal stones.Wide-focus high energy lithotripsy was associated with lower re-treat rate and higher complication rate for stones more than 10 mm.NGAL may play a potential role in the evalua-tion of SWL induced early renal injury.