Immunomodulation Effect of Kaempferia galanga Ethanolic Extract On In Vitro Lymphocyte Cell Proliferation

<p><em>Kaempferia galanga</em> L. is a traditional medicine with antitumor properties, as indicated by its immunomodulatory activities. This study aimed to determine the effect of <em>K. galanga</em> on lymphocyte cell proliferation activity as an indicator of immunomodulatory properties. This study was conducted at the Indonesian Research Center for Veterinary Science (February to April 2018). The immunomodulatory activity of the extract was evaluated with an <em>in vitro</em> splenocyte proliferation assay. The assay was based on cellular enzymatic synthesis to transform the XTT from formazan tetrazolium as an indicator. The <em>K. galanga</em> extract was obtained by 96% ethanol extraction. The test was conducted in an aseptic condition, consisted of five treatment groups with three replications each. Three groups of splenocyte cell culture, each with extract concentration of 2.5 µg.ml^-1, 25 µg.ml^-1, and 250 µg.ml^-1, as well as a positive (Concanavalin A/Con A) and negative (cell only) control. The cell suspension (10x10⁴cells/ml) was distributed on 96-well plates and cultured following the treatment groups. The same five plates were made for five days of observation and retrieved daily by observing an Elisa reader at 450 nm. The extract of <em>K. galanga </em>at 2.5 µg.ml^-1, 25 µg.ml^-1, and 250 µg.ml^-1 significantly (P &lt;0.05) promoted splenocyte proliferation compared to control. Therefore, it was expected that <em>K. galanga </em>has a high potential to be used as immunomodulators. Hence, further investigations should be done to clarify the mechanisms of the immunomodulatory effect of <em>K. galanga</em> as an antitumor <em>in vivo</em>.</p>

Probable Causes and Risk Factors for Positive SARS-CoV-2 Testing in Recovered Patients: Evidence From Guangzhou, China

Some patients retested positive for SARS-CoV-2 following negative testing results and discharge. However, the potential risk factors associated with redetectable positive testing results in a large sample of patients who recovered from COVID-19 have not been well-estimated. A total of 745 discharged patients were enrolled between January 30, 2020, and September 9, 2020, in Guangzhou, China. Data on the clinical characteristics, comorbidities, drug therapy, RT-PCR testing, and contact modes to close contacts were collected. Patients who tested positive for SARS-CoV-2 after discharge were confirmed by guidelines issued by China. The repositive rate in different settings was calculated. Among 745 discharged patients, 157 (21.1%; 95% CI, 18.2–24.0%) tested repositive and the repositive rate was 16.8% (95% CI, 14.1–24.0%) for nasopharyngeal swabs and 9.7% (95% CI, 7.0–12.5%) for anal swabs. Among them, 55 (35.0%) were asymptomatic, 15 (9.6%) had mild symptoms, 83 (52.9%) had moderate symptoms, and 4 (2.6%) had severe symptoms at the first admission. The days from discharge to repositivity was 8.0 (IQR, 8.0–14.0). Most repositive patients were without clinical symptoms, and lymphocyte cell counts were higher than before being discharged. The likelihood of repositive testing for SARS-CoV-2 RNA was significantly higher among patients who were of younger age (OR, 3.88; 95% CI, 1.74–8.66, 0–17 years old), had asymptomatic severity (OR, 4.36; 95% CI, 1.47–12.95), and did not have clinical symptoms (OR, 1.89; 95% CI, 1.32–2.70, without fever). No other positive patients emerged within the families or close contacts of repositive patients. Our findings support prolonged but intermittent viral shedding as the probable cause for this phenomenon; we need to familiarize with the possibility that the virus will remain endemic.

Identification of lymphocyte cell-specific protein-tyrosine kinase (LCK) as a driver for invasion and migration of oral cancer by tumor heterogeneity exploitation

Background Cancer metastases are the main cause of lethality. The five-year survival rate for patients diagnosed with advanced stage oral cancer is 30%. Hence, the identification of novel therapeutic targets is an urgent need. However, tumors are comprised of a heterogeneous collection of cells with distinct genetic and molecular profiles that can differentially promote metastasis making therapy development a challenging task. Here, we leveraged intratumoral heterogeneity in order to identify drivers of cancer cell motility that might be druggable targets for anti-metastasis therapy. Methods We used 2D migration and 3D matrigel-based invasion assays to characterize the invasive heterogeneity among and within four human oral cancer cell lines in vitro. Subsequently, we applied mRNA-sequencing to map the transcriptomes of poorly and strongly invasive subclones as well as primary tumors and matched metastasis. Results We identified SAS cells as a highly invasive oral cancer cell line. Clonal analysis of SAS yielded a panel of 20 subclones with different invasive capacities. Integrative gene expression analysis identified the Lymphocyte cell-specific protein-tyrosine kinase (LCK) as a druggable target gene associated with cancer cell invasion and metastasis. Inhibition of LCK using A-770041 or dasatinib blocked invasion of highly aggressive SAS cells. Interestingly, reduction of LCK activity increased the formation of adherens junctions and induced cell differentiation. Conclusion Analysis of invasive heterogeneity led to the discovery of LCK as an important regulator of motility in oral cancer cells. Hence, small molecule mediated inhibition of LCK could be a promising anti-metastasis therapy option for oral cancer patients.

Human immunodeficiency virus-related renal cell carcinoma: a retrospective study of 19 cases

Purpose We aimed to investigate basic information, clinical findings, treatments for tumor, pathology, and outcomes of HIV-positive patients diagnosed with renal cell carcinoma (RCC). Patients and methods We collected 19 patients from 2012 to 2020 who are diagnosed with RCC with HIV-positive. A retrospective analysis was performed on their hospitalization course and tumor-related parameters, including basic information, clinical findings, HIV-associated data, pathology, treatments for tumor, and outcomes. Results In our study, patients were diagnosed with RCC at the median age of 51. Males took a great part (17 males, 89%) in all patients, while only 2 females were diagnosed. The median CD4+ T lymphocyte cell count was 462 cells/μl when diagnosed with RCC (range from 111 cells/μl to 1536 cells/μl). Eleven patients diagnosed with RCC and HIV infection at the same time, who may have high viral load and low CD4+ T lymphocyte cell count. Eight patients accepted a median HAART for 30 months (range from 11 months to 108 months) prior to diagnosis of RCC. All the patients performed operations successfully, and 4 of them performed partial nephrecotomy. Only 1 patient was identified with chromophobe cell carcinoma, 1 with partially clear cell and partially papillary carcinoma, and 17 with clear cell carcinoma. Two of the patients with Fuhrman grades 2–3 accepted cytokine therapy with IL-2 and IFN-α. Two patients died of lung metastasis 1 year and 6 months after surgery respectively, even though 1 patient accepted full dose targeted therapy (sorafenib) for 3 months, and one refused adjuvant therapy. The remaining 17 patients are still alive at a median follow-up of 34 months; however, 1 patient lives with lung and brain metastases at the last follow-up of 3 years after surgery. Conclusions RCC patients with HIV-positive were similar to the general population in terms of clinical characters, treatment measures, and pathology. RCC patients with HIV-positive seemed like to obey the same clinical practice guideline as in the general population. The outcomes of HIV-positive patients with partial nephrectomy are not inferior to patients with radical nephrectomy. Furthermore, experience in targeted therapy and immunal therapy (PD-1/PD-L1 inhibitors) needs to be learned.

System-wide hematopoietic and immune signaling aberrations in COVID-19 revealed by deep proteome and phosphoproteome analysis

The COVID-19 pandemic, caused by the SARS-CoV-2 virus, has become a global crisis. To gain systems-level insights into its pathogenesis, we compared the blood proteome and phosphoproteome of ICU patients with or without SARS-CoV-2 infection, and healthy control subjects by quantitative mass spectrometry. We find that COVID-19 is marked with hyperactive T cell and B cell signaling, compromised innate immune response, and dysregulated inflammation, coagulation, metabolism, RNA splicing, transcription and translation pathways. SARS-CoV-2 infection causes global reprogramming of the kinome and kinase-substrate network, resulting in defective antiviral defense via the CK2-OPN-IL-12/IFN-I axis, lymphocyte cell death via aberrant JAK/STAT signaling, and inactivation of innate immune cells via inhibitory SIRPA, SIGLEC and SLAM family receptor signaling. Our work identifies CK2, SYK, JAK3, TYK2 and IL-12 as potential targets for immunomodulatory treatment of severe COVID-19 and provides a valuable approach and resource for deciphering the mechanism of pathogen-host interactions.

Human Immunodeficiency Virus Related Renal Cell Carcinoma: a Retrospective Study of 19 Cases

Purpose: We aimed to investigate basic information, clinical findings, treatments for tumor, pathology and outcomes of HIV-positive patients diagnosed renal cell carcinoma(RCC). Patients and Methods: We collected 19 patients from 2012 to 2020 who diagnosed RCC with HIV- positive. A retrospective analyze was performed on their hospitalization course and tumor related parameters, including basic information, clinical findings, HIV associated datas, pathology, treatments for tumor and outcomes.Results: In our study, patients were diagnosed RCC at the median age 51. Males took a great part(17 males,89%) in all patients, while only 2 females were diagnosed. The median CD4+ T-lymphocyte cell count was 462 cells/ul when diagnosed RCC(range from 111 cells/ul to 1536 cells/ul). 11 patients diagnosed RCC and HIV infection at the same time, who may have high viral load and low CD4+ T-lymphocyte cell count. 8 patients accepted a median HAART for 30 months(range from 11 months to 108months) prior to diagnosis of RCC. All the patients performed operations successfully, and 4 of them performed partial nephrecotomy. Only 1 patient was identified with chromophobe cell carcinoma, 1 with partially clear cell and partialy papillary carcinoma, and 17 with clear cell carcinoma. 2 of the patients with Fuhrman grade2-3 accepted cytokine therapy with IL-2 and IFN-α. 2 patients died of lung metastasis one year and six months after surgery respectively, even though 1 patient accepted full dose targeted therapy (Sorafenib) for 3 months, and one refused adjuvant therapy. The remaining 17 patients are still alive at a median follow-up of 34 months, however 1 patient live with lung and brain metastases at last follow-up of 3 years after surgery.Conclusions: RCC Patients with HIV-positive were similar to general population in terms of clinical characters, treatment measures, and pathology. RCC patients with HIV- positive seemed like to obey the same clinical practice guideline as in general population. The outcomes of HIV-positive patients with partial nephrectomy are not inferior to patients with radical nephrectomy. Further more experience in targeted therapy and immunal therapy( PD-1/ PD-L1 inhibitors) needs to be learned.

System-wide hematopoietic and immune signaling aberrations in COVID-19 revealed by deep proteome and phosphoproteome analysis

The COVID-19 pandemic, caused by the SARS-CoV-2 virus, has become a global crisis. To gain systems-level insights into its pathogenesis, we compared the blood proteome and phosphoproteome of ICU patients with or without SARS-CoV-2 infection, and healthy control subjects by quantitative mass spectrometry. We find that COVID-19 is marked with hyperactive T cell and B cell signaling, compromised innate immune response, and dysregulated inflammation, coagulation, metabolism, RNA splicing, transcription and translation pathways. SARS-CoV-2 infection causes global reprogramming of the kinome and kinase-substrate network, resulting in defective antiviral defense via the CK2-OPN-IL-12/IFN-I axis, lymphocyte cell death via aberrant JAK/STAT signaling, and inactivation of innate immune cells via inhibitory SIRPA, SIGLEC and SLAM family receptor signaling. Our work identifies CK2, SYK, JAK3, TYK2 and IL-12 as potential targets for immunomodulatory treatment of severe COVID-19 and provides a valuable approach and resource for deciphering the mechanism of pathogen-host interactions.