A New Auto-RPA-Fluorescence Detection Platform for SARS-CoV-2.

OBJECTIVE: The outbreak of COVID-19 caused by SARS-CoV-2 has led to a serious worldwide pandemic. Quantitative reverse transcription-polymerase chain reaction (qRT-PCR)-based methods were recommended for routine detection of SARS-CoV-2 RNA. Because the reaction time and analytical sensitivity of qRT-PCR limits the diagnosis of SARS-CoV-2, development of a quick process of SARS-CoV-2 detection technology with high analytical sensitivity remains urgent. METHODS: We combined isothermal amplification and fluorescence detection technology to develop a new auto-recombinase polymerase amplification (RPA)-fluorescence platform that could be used in the diagnosis of SARS-CoV-2. RESULTS: By optimization of primers and probes, the RPA platform could detect SARS-CoV-2 nucleotides within 15 min. The limits of detection and specificity of the auto-RPA-fluorescence platform were 5 copies/µL and 100%, respectively. The accuracy of detection of the auto-RPA-fluorescence platform in the 16 positive samples was 100%. CONCLUSION: The RPA platform is a potential technology for the diagnosis of SARS-CoV-2 infection.

Shh Overexpression Is Correlated with GRP78 and AR Expression in Primary Prostate Cancer: Clinicopathological Features and Outcomes in a Chinese Cohort.

INTRODUCTION: Shh plays an important role in prostate cancer progression, but its correlation with GRP78 and AR is elusive. METHODS: The study included 539 patients in total, of which 443 had primary prostate carcinoma and 96 patients had benign prostatic hyperplasia (BPH). The clinicopathologic features, histologic scores of protein expression, and correlations between protein and disease state were studied in this cohort. Kaplan-Meier and Pearson correlation analyses were used to compare measures between groups. We performed immunohistochemistry to evaluate the expression of the Shh protein in benign prostatic hyperplasia (n=96) and prostate cancer (Gleason scores ≤6 [n=399] or ≥7 [n=44]). We quantified the expression of Shh, AR, and GRP78 using the weighted histoscore method, studied the correlation between Shh expression and AR and GRP78, and evaluated the impact of Shh protein expression on patient survival. RESULTS: Shh expression was significantly higher in prostate cancer with Gleason scores ≥7 than in cancer with lower Gleason scores or benign hyperplasia and was much higher in AR-positive cancer than in AR-negative cancer. Shh is overexpressed in high-grade prostate cancer and is positively correlated with the expression of both GRP78 and AR. CONCLUSION: Therefore, Shh may be a useful prognostic marker and therapeutic target for prostate cancer.

Quercetin Enhanced Paclitaxel Therapeutic Effects Towards PC-3 Prostate Cancer Through ER Stress Induction and ROS Production.

INTRODUCTION: Prostate cancer is one of the most common cancers threatening public health worldwide. Although chemotherapy plays an important role in treating prostate cancer, it leads to many adverse effects and is prone to drug resistance. Quercetin, a natural product, is used in traditional Chinese medicine because of its strong antitumor activity and few side effects. METHODS: In this study, we combined quercetin and paclitaxel to kill prostate cancer cells in vivo and in vitro, and we investigated the relevant mechanism of this combination treatment. After the cancer cells were treated with quercetin or/and paclitaxel, cell growth inhibition, apoptosis, the cell cycle, reactive oxygen species (ROS) generation, and several endoplasmic reticulum (ER) stress signaling pathway related gene expressions were evaluated. RESULTS: The combined treatment with quercetin and paclitaxel significantly inhibited cell proliferation, increased apoptosis, arrested the cell cycle at the G2/M phase, inhibited cell migration, dramatically induced ER stress to occur, and increased ROS generation. In a PC-3 cancer-bearing murine model, this combination treatment exerted the most beneficial therapeutic effects, and quercetin increased the cancer cell-killing effects of paclitaxel, with nearly no side effects compared with the single paclitaxel treatment group. CONCLUSION: Combination treatment possessed enhanced anti-cancer effects, and these results will provide a basis for treating prostate cancer using a combination of quercetin and paclitaxel.

Codelivery of GRP78 siRNA and docetaxel via RGD-PEG-DSPE/DOPA/CaP nanoparticles for the treatment of castration-resistant prostate cancer.

Background: Castration-resistant prostate cancer (CRPC) accounts for the majority of prostate cancer deaths, and patients with CRPC are prone to developing drug resistance. Therefore, there is a need to develop effective therapeutics to treat CRPC, especially drug-resistant CRPC. Although various nanoparticles have been developed for drug or gene delivery and control release, approaches to reproducibly formulate the optimal treatment with nanoparticles that could effectively target CRPC and bone metastasis remain suboptimal. Recently, codelivery of a chemotherapeutic agent and a small interfering RNA (siRNA) has become a promising strategy for the treatment of drug-resistant prostate cancer. Methods: In a previous study, we prepared a novel RGD-PEG-DSPE/CaP nanoparticle as an effective and biocompatible drug and gene delivery system. In this study, we further modify the nanoparticle to obtain the LCP-RGD nanoparticle, which contains a calcium phosphate (CaP) core, dioleoyl phosphatidic acid (DOPA) and RGD modified poly(ethylene glycol)-conjugated distearoyl phosphatidylethanolamine (RGD-PEG-DSPE). This drug delivery system was used for codelivery of GRP78 siRNA and docetaxel (DTXL) for the treatment of the PC-3 CRPC. Results: The nanoparticles contain the CaP core, which can effectively compress the negatively charged siRNA, while the DOPA and RGD-PEG-DSPE component can effectively carry DTXL. The arginine-glycine-aspartic acid (RGD) segment can target the prostate cancer site, as the cancer site is neovascularized. This novel nanoparticle has good stability, excellent biocompatibility, high drug and siRNA loading capacity, and an in vitro sustainable release profile. Conclusion: Codelivery of DTXL and GRP78 siRNA has enhanced in vitro and in vivo anti-prostate cancer effects which are much greater than using free DTXL and free GRP78 siRNA together. Our study also indicated that codelivery of DTXL and GRP78 siRNA have an in vitro and in vivo combinational anti-prostate cancer effect and also could effectively sensitize the cell-killing effect of DTXL; this method may be especially suitable for drug-resistant CRPC treatment.

Clinicopathologic Characteristics of Thyroid-like Low-grade Nasopharyngeal Papillary Adenocarcinoma: A Case Report.

Thyroid-like low-grade nasopharyngeal papillary adenocarcinoma (TL-LGNPPA) is an extremely rare neoplasm. It morphologically resembles papillary thyroid carcinoma and is immunochemically positive for thyroid transcription factor 1. Herein, we reported a case of TL-LGNPPA in a female patient of 35 years old. She felt pharyngeal foreign body sensation of unknown cause and subsequently developed dyspnea after activities. Nasopharyngeal plain computerized tomography showed no abnormalities, but laryngoscopy showed a peanut-sized smooth neoplasm with a pedicel at the posterior edge of the nasal septum. The mass was completely resected by nasopharyngoscopy. Histologic examination showed the tumor was composed of papillary configuration and tubular glands; each papilla was covered with cuboidal or columnar epithelial cells; tubular architecture and spindle cell component were also observed; some tumor cells had psammoma bodies. Immunohistochemically, tumor cells were positive for CKpan, CK7, CK19, VIM, and thyroid transcription factor 1, but negative for thyroglobulin, CK20, S-100, P63, P40, smooth muscle actin, CDX-2, and glial fibrillary acidic protein. This patient was diagnosed with TL-LGNPPA and followed up for 16 months, and metastasis and recurrence were not observed.

Hepatocellular carcinoma metastasis to the mandibular ramus: a case report.

Hepatocellular carcinoma (HCC) is one of the most commonly reported cancers worldwide. Although the extrahepatic metastasis of this carcinoma has been reported frequently, the mandible is a rare site for HCC metastasis. Here we have presented a rare case of mandibular metastatic HCC in a 37-year-old man presenting with painless swelling over his right parotid gland for the previous 3 months. This patient had a history of HCC caused by a hepatitis C viral infection. The tumor cells exhibited eosinophilic and rich cytoplasm, and they appeared to be trabecular (similar to hepatic cells), with sinusoids between the tumor cells. Moreover, the tumor cells were positive for Hep Par-1 and Glypican-3 in the cytoplasm. A 64-multidetector-row computed tomography scan was performed, and it showed pathological tracer uptake in the right mandible. Therefore, an excisional biopsy was performed, and with the support of an immunohistochemical analysis, an HCC mandibular metastasis diagnosis was made. The patient was treated with radiotherapy and chemotherapy. This patient was followed up in the oncology department for 1 year, and he was still alive at the time this report was written. This extremely rare case highlights the differential diagnosis difficulties that can be encountered, and the importance of diagnostic clues, such as the clinical history, histopathology, and immunohistochemistry, in the establishment of a definitive diagnosis. Clinicians should be aware of the possibility of encountering HCC metastasis in the mandible, and take this into consideration in order to prevent a misdiagnosis.

Efficacy and toxicities of combination maintenance therapy in the treatment of advanced colorectal cancer: a meta-analysis of randomized controlled trials.

PURPOSE: We performed a systematic review and meta-analysis to investigate the efficacy and toxicities of combination maintenance therapy for the treatment of advanced colorectal cancer (CRC). METHODS: Relevant trials were identified by searching electronic databases and conference meetings. Prospective randomized controlled trials (RCTs) assessing combination maintenance therapy in advanced CRC patients were included. Outcomes of interest included overall survival (OS), progression-free survival (PFS) and grade 3-4 toxicities. RESULTS: A total of 3,174 advanced CRC patients received combination maintenance treatment from 6 RCTs were included for analysis. The use of combination maintenance therapy did not significantly improved PFS (HR 0.95, 95%CI: 0.75-1.20, p =0.67) and OS (HR 1.05, 95%CI: 0.93-1.17, p =0.45) in comparison with single bevacizumab maintenance therapy for the treatment of advanced CRC, similar results were observed in sub-group analysis according to treatment regimens. In addition, combination maintenance therapy significantly improved PFS (HR 0.57, 95%CI: 0.41-0.80, p =0.001), but not for OS (HR 0.93, 95%CI: 0.76-1.14, p =0.47) in comparison with observation. Additionally, more incidences of any grade 3-4 toxicities (diarrhea, fatigue and hand-foot skin reaction) were observed in the combination maintenance therapy. CONCLUSIONS: The findings of this study show that the efficacy of combination maintenance therapy is comparable to that of bevacizumab alone in terms of PFS and OS for advanced CRC patients, but at the cost of increased grade 3-4 toxicities. Thus single agent bevacizumab remains the recommended maintenance treatment for advanced CRC patients.

Sigma-1 receptor (σ1R) is downregulated in hepatic malignant tumors and regulates HepG2 cell proliferation, migration and apoptosis.

Sigma-1 receptor (σ1R), an important transmembrane structural protein, has been demonstrated to be overexpressed in various types of human cancer, and has been confirmed to be involved in many biological behaviors during tumorigenesis and tumor progression. The aim of the present study was to explore the essential role of σ1R in hepatic malignant tumors (HMTs), which, to the best of our knowledge, has not been reported to date. We assessed σ1R expression in hepatocellular carcinoma (HCC) tissues and found that σ1R was significantly decreased in HCC when compared with that in benign liver tissues (P<0.01). Additionally, the expression of σ1R was shown to be inversely correlated with HCC grade (r=-0.424, P=0.021, Kendall's τ-b-test). We further used a FLAG­SV40­neomycin­plasmid strategy to increase σ1R expression in the HepG2 hepatoblastoma cell line. Overexpression of σ1R impaired cell proliferation, inhibited cell migration, induced cell cycle arrest at G1 phase, and increased cell apoptosis in vitro. Furthermore, overexpression of σ1R decreased the expression levels of STAT-3 and NF-κB, which provided insight into the underlying mechanisms of σ1R-associated HMT development and progression. These findings suggest that the decreased expression of σ1R plays an essential role in hepatic tumorigenesis, and that it may serve as a potential predictive factor and therapeutic target for the treatment of HMTs.

Clinical and prognostic significance of SOX11 in breast cancer.

Recently, the transcription factor SOX11 has gained extensive attention as a diagnostic marker in a series of cancers. However, to date, the possible roles of SOX11 in breast cancer has not been investigated. In this study, immunohistochemical staining for SOX11 was performed for 116 cases of breast cancer. Nuclear SOX11 was observed in 42 (36.2%) and cytoplasmic SOX11 in 52 (44.8%) of breast cancer samples. Moreover, high expression of cytoplasmic and nuclear SOX11 was associated with clinicopathological factors, including earlier tumor grade, absence of lymph node metastasis and smaller tumor size. Kaplan-Meier survival curves demonstrated high nuclear SOX11 expression to be associated with more prolonged overall survival than those with low expression and it could be an independent predictor of survival for breast cancer patients. It is worthwhile to note that cytoplasmic SOX11 was not correlated with prognosis of breast cancer patients. These data suggest the possibility that nuclear SOX11 could be as a potential target for breast cancer therapy.