Discovery of selective and potent USP22 inhibitors via structure-based virtual screening and bioassays exerting anti-tumor activity.

Ubiquitin-specific protease 22 (USP22) plays a prominent role in tumor development, invasion, metastasis and immune reprogramming, which has been proposed as a potential therapeutic target for cancer. Herein, we employed a structure-based discovery and biological evaluation and discovered that Rottlerin (IC50 = 2.53 µM) and Morusin (IC50 = 8.29 µM) and as selective and potent USP22 inhibitors. Treatment of HCT116 cells and A375 cells with each of the two compounds resulted in increased monoubiquitination of histones H2A and H2B, as well as reduced protein expression levels of Sirt1 and PD-L1, all of which are known as USP22 substrates. Additionally, our study demonstrated that the administration of Rottlerin or Morusin resulted in an increase H2Bub levels, while simultaneously reducing the expression of Sirt1 and PD-L1 in a manner dependent on USP22. Furthermore, Rottlerin and Morusin were found to enhance the degradation of PD-L1 and Sirt1, as well as increase the polyubiquitination of endogenous PD-L1 and Sirt1 in HCT116 cells. Moreover, in an in vivo syngeneic tumor model, Rottlerin and Morusin exhibited potent antitumor activity, which was accompanied by an enhanced infiltration of T cells into the tumor tissues. Using in-depth molecular dynamics (MD) and binding free energy calculation, conserved residue Leu475 and non-conserved residue Arg419 were proven to be crucial for the binding affinity and inhibitory function of USP22 inhibitors. In summary, our study established a highly efficient approach for USP22-specific inhibitor discovery, which lead to identification of two selective and potent USP22 inhibitors as potential drugs in anticancer therapy.

The Impact of Chemosensitivity on the Outcome of Brain Metastases in Small-Cell Lung Cancer: A Retrospective Analysis.

PURPOSE: The purpose of this study was to investigate the prognostic differences between patients with small-cell lung cancer (SCLC) with different chemosensitivity to first-line chemotherapy who developed brain metastasis (BM) as the first site of progression. METHODS: Patients with a BM after first-line treatment in the Tianjin Cancer Hospital were retrospectively analyzed. According to the time-free interval (TFI) between the completion of first-line chemotherapy and the onset of the BM, the patients were divided into the chemo-sensitive group (TFI ≥ 90 days, n = 145) and the chemo-resistant group (TFI < 90 days, n = 97). The survival time, which was calculated from the diagnosis of the BM, was analyzed after the onset of brain metastasis (BM-OS). Survival curves were plotted using the Kaplan-Meier method, and differences between groups were compared using the log-rank test. RESULTS: In total, the median BM-OS was 8.4 months. The median BM-OS in the chemo-sensitive group was 8.8 months, and it was 8.0 months in the chemo-resistant group (p = 0.538). In patients without extracranial progression (n = 193), the median BM-OSes in the chemo-sensitive and chemo-resistant groups were 9.4 months and 9.7 months, respectively (p = 0.947). In patients with extracranial progression (n = 49), the median BM-OSes in the chemo-sensitive and chemo-resistant groups were 5.4 months and 4.2 months, respectively (p = 0.161). Conclusions: After the development of a BM as the first site of progression following chemotherapy in patients with SCLC, the prognosis of chemo-sensitive patients was not necessarily superior to chemo-resistant patients, especially in patients without extracranial progression.

Efficacy and safety of EGFR inhibitors and radiotherapy in locally advanced non-small-cell lung cancer: a meta-analysis.

Aim: To assess the efficacy and safety of EGFR inhibitors combined with (chemo)radiotherapy in unresectable, locally advanced non-small-cell lung cancer. Materials & methods: A systematic review and meta-analysis of prospective trials was performed. Results: Twenty-eight studies of 1640 patients were included. In patients harboring EGFR-sensitive mutations, the pooled objective response rate, 1-year overall survival rate and 1-year progression-free survival rate of EGFR-TKIs + (chemo)radiotherapy were 0.803, 0.766 and 0.554, respectively. Compared with chemoradiotherapy, the addition of EGFR inhibitors did not significantly increase the risk of grade ≥3 pneumonitis and esophagitis. Conclusion: EGFR-tyrosine kinase inhibitors combined with (chemo)radiotherapy are tolerable and the clinical benefit is promising, especially in patients with EGFR-sensitive mutations.

The aim of this systemic review and meta-analysis was to assess the efficacy and safety of (chemo)radiotherapy combined with therapies targeting EGFR receptor, in unresectable, locally advanced non-small-cell lung cancer. Prospective clinical trials were searched and analyzed, and 28 studies of 1640 patients were included in this analysis. The results showed that the efficacy of (chemo)radiotherapy combined with tyrosine kinase inhibitors targeting EGFR, such as gefitinib and erlotinib, was promising, especially among patients harboring sensitive mutations in EGFR. Besides, this combination therapy was safe, which did not increase the risk of severe pneumonitis and esophagitis. Overall, tyrosine kinase inhibitors targeting EGFR combined with (chemo)radiotherapy are tolerable and the clinical benefit is promising, especially in patients with EGFR-sensitive mutations.

GATA6 activated long non-coding RNA PCAT1 maintains stemness of non-small cell lung cancer by mediating FRK.

PURPOSE: Long non-coding RNA (lncRNA) prostate cancer-associated transcripts 1 (PCAT1) is a noticeable lncRNA involved in the tumorigenesis of various cancers. Nowadays, the biological function of PCAT1 on the stemness of non-small cell lung cancer (NSCLC) is still unclear. Our purpose was to explore the molecular mechanism of PCAT1 and its target protein in advanced NSCLC. METHODS: The levels of PCAT1 and Fyn-related kinase (FRK) in NSCLC tissues and cell lines were evaluated by quantitative polymerase chain reaction (qPCR). The log-rank test was applied to evaluate the role of high PCAT1 levels in shortening the overall survival of NSCLC patients. Chi-square test was to assess the relation between PCAT1 expression and clinicopathological features of NSCLC patients. CCK8 assay tested the cell proliferation of NSCLC cells with PCAT1 overexpression. The underlying regulatory mechanism between PCAT1 and Fyn-related kinase (FRK) was predicted by bioinformatics and verified by RNA transfection, qPCR, and western blotting. Chromatin immunoprecipitation (ChIP) assay was done to examine the relation between GATA binding protein 6 (GATA6) and the PACT1 gene. Mice xenografts were applied to examine the function of PACT1 on NSCLC development in vivo. RESULTS: PCAT1 was aberrantly elevated in tissues from patients with NSCLC. High levels of PCAT1 expression were more likely to present in patients with late-stage, positive CD133, and inferior overall survival. PCAT1 knockdown reduced cell proliferation, stem cell properties (Sox2 and Nanog expression) of H226 and A549 cells in vitro, and repressed tumor development in vivo. Furthermore, FRK in NSCLC cells was downregulated by silencing PCAT1. The tissue level of FRK was positively correlated with PCAT1 expression in patients with NSCLC. Furthermore, GATA6 was found to be promoter of PCAT1 and increased PCAT1 levels in NSCLC cells. CONCLUSIONS: GATA6/PCAT1 may markedly maintain the stemness of NSCLC, resulting in late TNM stage and poor survival. These findings suggest that the GATA6-PCAT1-FRK axis may be a useful target for intervention in NSCLC.

Deep Vein Thrombosis in Hospitalized Patients With COVID-19 in Wuhan, China: Prevalence, Risk Factors, and Outcome.

BACKGROUND: To investigate deep vein thrombosis (DVT) in hospitalized patients with coronavirus disease 2019 (COVID-19), we performed a single institutional study to evaluate its prevalence, risk factors, prognosis, and potential thromboprophylaxis strategies in a large referral and treatment center. METHODS: We studied a total of 143 patients with COVID-19 from January 29, 2020 to February 29, 2020. Demographic and clinical data, laboratory data, including ultrasound scans of the lower extremities, and outcome variables were obtained, and comparisons were made between groups with and without DVT. RESULTS: Of the 143 patients hospitalized with COVID-19 (age 63±14 years, 74 [51.7%] men), 66 patients developed lower extremity DVT (46.1%: 23 [34.8%] with proximal DVT and 43 [65.2%] with distal DVT). Compared with patients who did not have DVT, patients with DVT were older and had a lower oxygenation index, a higher rate of cardiac injury, and worse prognosis, including an increased proportion of deaths (23 [34.8%] versus 9 [11.7%]; P=0.001) and a decreased proportion of patients discharged (32 [48.5%] versus 60 [77.9%]; P<0.001). Multivariant analysis showed an association only between CURB-65 (confusion status, urea, respiratory rate, and blood pressure) score 3 to 5 (odds ratio, 6.122; P=0.031), Padua prediction score ≥4 (odds ratio, 4.016; P=0.04), D-dimer >1.0 µg/mL (odds ratio, 5.818; P<0.014), and DVT in this cohort, respectively. The combination of a CURB-65 score 3 to 5, a Padua prediction score ≥4, and D-dimer >1.0 µg/mL has a sensitivity of 88.52% and a specificity of 61.43% for screening for DVT. In the subgroup of patients with a Padua prediction score ≥4 and whose ultrasound scans were performed >72 hours after admission, DVT was present in 18 (34.0%) patients in the subgroup receiving venous thromboembolism prophylaxis versus 35 (66.0%) patients in the nonprophylaxis group (P=0.010). CONCLUSIONS: The prevalence of DVT is high and is associated with adverse outcomes in hospitalized patients with COVID-19. Prophylaxis for venous thromboembolism may be protective in patients with a Padua protection score ≥4 after admission. Our data seem to suggest that COVID-19 is probably an additional risk factor for DVT in hospitalized patients.

Time-effect of penile color duplex Doppler ultrasound for diagnosing vascular erectile dysfunction.

INTRODUCTION: This study aimed to explore the time-effect of color duplex Doppler ultrasound (CDDU) in the diagnosis of vascular erectile dysfunction (ED). MATERIAL AND METHODS: Using a self-control study, we included patients who underwent penile CDDU and cavernosography in our hospital. We compared the arterial peak systolic velocity (PSV) of CDDU among different intervals for the diagnosis of arterial ED. We included 357 patients who were under consideration for vascular ED. RESULTS: We found significant differences in all the pairwise comparison of PSV in the 1st (0-5 min), 2nd (6-10 min), 3rd (11-15 min), and 4th (16-20 min) 4 intervals after the injection of prostaglandin E1 (p<0.001), except the 11-15 min vs. the 16-20 min interval (p=0.387). Using cavernosography, 294 patients were diagnosed with venous ED. Compared with other intervals, the diagnosis of CDDU 11-15 min after the intracavernous injection of 20 ug prostaglandin E1 (PGE1) had the best consistency with cavernosography (Kappa=0.761; p<0.001). Compared with other intervals, CDDU at 11-15 min had the highest specificity (93.65%), the highest Youden index (0.85), positive likelihood ratio of 14.46, positive predictive value of 98.54%, negative predictive value of 71.08% and a coincidence rate of 92.16%. CONCLUSIONS: Our findings support the increased utilization of CDDU for the diagnosis of both arterial and venous ED. The diagnosis at 11-15 min after intracavernous injection of PGE1 is accurate and stable, which would help to simplify the process and shorten the time of CDDU.

Diagnostic Accuracy of Different Criteria of Pharmaco-penile Duplex Sonography for Venous Erectile Dysfunction.

OBJECTIVES: The aim of this study was to analyze the diagnostic accuracy of different criteria of pharmaco-penile duplex sonography in venous erectile dysfunction (ED). METHODS: The following parameters were measured after an intracavernous injection test in patients with ED from May 2016 to February 2017 at our hospital: diameter, peak systolic velocity, end-diastolic velocity, and resistance index of the cavernous artery; diameter and peak velocity (if leak occurred) of the deep dorsal vein. Three ultrasonographic diagnostic criteria of venous ED were applied. Criterion A: continuous blood flow signals in the deep dorsal vein, peak velocity greater than 3 cm/s, peak systolic velocity greater than 30 cm/s, end-diastolic velocity greater than 5 cm/s; Criterion B: resistance index less than 0.89 and other parameters corresponding with Criterion A; Criterion C: resistance index less than 0.80 and other parameters corresponding with Criterion A. The diagnostic results of each criterion were compared with the cavernosographic results. RESULTS: Thirty-six patients were diagnosed as venous ED by cavernosography in 54 ED cases. The diagnostic specificity, sensitivity, and accuracy of Criterion A were 70.6%, 91.7%, and 84.9%, respectively. Those of Criterion B were 82.4%, 69.4%, and 73.6%, while the results for Criterion C were 94.1%, 33.3%, and 52.8%, respectively. Criterion A had the highest diagnostic accuracy, the largest area under the receiver operating characteristic curve (area = 0.811), and the highest consistency (kappa = 0.642) with the cavernosographic results in the 3 criteria. The difference was statistically significant (P < .05). CONCLUSIONS: Among the 3 commonly used ultrasonographic criteria, Criterion A is most appropriate in the diagnosis of venous ED.

Ammonium removal using algae-bacteria consortia: the effect of ammonium concentration, algae biomass, and light.

In this study, the effects of ammonium nitrogen concentration, algae biomass concentration, and light conditions (wavelength and intensity) on the ammonium removal efficiency of algae-bacteria consortia from wastewater were investigated. The results indicated that ammonium concentration and light intensity had a significant impact on nitrification. It was found that the highest ammonia concentration (430 mg N/L) in the influent resulted in the highest ammonia removal rate of 108 ± 3.6 mg N/L/days, which was two times higher than the influent with low ammonia concentration (40 mg N/L). At the lowest light intensity of 1000 Lux, algae biomass concentration, light wavelength, and light cycle did not show a significant effect on the performance of algal-bacterial consortium. Furthermore, the ammonia removal rate was approximately 83 ± 1.0 mg N/L/days, which was up to 40% faster than at the light intensity of 2500 Lux. It was concluded that the algae-bacteria consortia can effectively remove nitrogen from wastewater and the removal performance can be stabilized and enhanced using the low light intensity of 1000 Lux that is also a cost-effective strategy.

Study on the effect of landfill leachate on nutrient removal from municipal wastewater.

In this study, landfill leachate with and without pre-treatment was co-treated with municipal wastewater at different mixing ratios. The leachate pre-treatment was achieved by air stripping to removal ammonia. The objective of this study was to investigate the effect of landfill leachate on nutrient removal of the wastewater treatment process. It was demonstrated that when landfill leachate was co-treated with municipal wastewater, the high ammonia concentration in the leachate did not have a negative impact on the nitrification. The system was able to adapt to the environment and was able to improve nitrification capacity. The readily biodegradable portion of chemical oxygen demand (COD) in the leachate was utilized by the system to improve phosphorus and nitrate removal. However, this portion was small and majority of the COD ended up in the effluent thereby decreased the quality of the effluent. The study showed that the 2.5% mixing ratio of leachate with wastewater improved the overall biological nutrient removal process of the system without compromising the COD removal efficiency.

Development of functional fish feed with natural ingredients to control heavy metals.

The effects of two natural ingredients, Chinese parsley (CP) and chitosan (CT), on growth, accumulation, and excretion of cadmium in fish body and preservation of essential trace metals in the body were investigated by using rainbow trout that had been fed cadmium-added diet, low and high concentration, for 3 weeks. This pretest confirmed that cadmium was accumulated in the liver, kidney, and intestine of the test fish. The cadmium level of the fish, fed diet with CP or CT, was decreased by 18% and 24%, respectively, compared to that of the fish given the control diet. But CP and CT did not have an influence on normal growth of test fish and the levels of essential trace metals in the body. In addition, the level of cadmium was higher in liver than kidney in the high-cadmium dietary group, indicating the Cd level in kidney follows that of liver as kidney lies in the final stage of metabolism. The cadmium accumulation in the fish body was supposed to be reduced, by giving CP to increase the solubility of Cd to body fluid by conjugation into metallothioneins (MTs), while CT was supposed to be responsible for the physical adsorption of cadmium ions by glucosamine groups.