Single-cell RNA sequencing revealed potential targets for immunotherapy studies in hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) is a solid tumor prone to chemotherapy resistance, and combined immunotherapy is expected to bring a breakthrough in HCC treatment. However, the tumor and tumor microenvironment (TME) of HCC is highly complex and heterogeneous, and there are still many unknowns regarding tumor cell stemness and metabolic reprogramming in HCC. In this study, we combined single-cell RNA sequencing data from 27 HCC tumor tissues and 4 adjacent non-tumor tissues, and bulk RNA sequencing data from 374 of the Cancer Genome Atlas (TCGA)-liver hepatocellular carcinoma (LIHC) samples to construct a global single-cell landscape atlas of HCC. We analyzed the enrichment of signaling pathways of different cells in HCC, and identified the developmental trajectories of cell subpopulations in the TME using pseudotime analysis. Subsequently, we performed transcription factors regulating different subpopulations and gene regulatory network analysis, respectively. In addition, we estimated the stemness index of tumor cells and analyzed the intercellular communication between tumors and key TME cell clusters. We identified novel HCC cell clusters that specifically express HP (HCC_HP), which may lead to higher tumor differentiation and tumor heterogeneity. In addition, we found that the HP gene expression-positive neutrophil cluster (Neu_AIF1) had extensive and strong intercellular communication with HCC cells, tumor endothelial cells (TEC) and cancer-associated fibroblasts (CAF), suggesting that clearance of this new cluster may inhibit HCC progression. Furthermore, ErbB signaling pathway and GnRH signaling pathway were found to be upregulated in almost all HCC tumor-associated stromal cells and immune cells, except NKT cells. Moreover, the high intercellular communication between HCC and HSPA1-positive TME cells suggests that the immune microenvironment may be reprogrammed. In summary, our present study depicted the single-cell landscape heterogeneity of human HCC, identified new cell clusters in tumor cells and neutrophils with potential implications for immunotherapy research, discovered complex intercellular communication between tumor cells and TME cells.

Insufficient tuberculosis treatment leads to earlier and higher mortality in individuals co-infected with HIV in southern China: a cohort study.

BACKGROUND: Tuberculosis (TB) and Acquired Immune Deficiency Syndrome (AIDS) are leading causes of death globally. However, little is known about the long-term mortality risk and the timeline of death in those co-infected with human immunodeficiency virus (HIV) and Mycobacterium tuberculosis (MTB). This study sought to understand the long-term mortality risk, factors, and the timeline of death in those with HIV-Mycobacterium tuberculosis (MTB) coinfection, particularly in those with insufficient TB treatment. METHODS: TB-cause specific deaths were classified using a modified 'Coding of Cause of Death in HIV' protocol. A longitudinal cross-registration-system checking approach was used to confirm HIV/MTB co-infection between two observational cohorts. Mortality from the end of TB treatment (6 months) to post-treatment year (PTY) 5 (60 months) was investigated by different TB treatment outcomes. General linear models were used to estimate the mean mortality at each time-point and change between time-points. Cox's proportional hazard regressions measured the mortality hazard risk (HR) at each time-point. The Mantel-Haenszel stratification was used to identify mortality risk factors. Mortality density was calculated by person year of follow-up. RESULTS: At the end point, mortality among patients with HIV/MTB coinfection was 34.7%. From the end of TB treatment to PTY5, mortality and loss of person years among individuals with TB treatment failure, missing, and adverse events (TBFMA) were significantly higher than those who had TB cure (TBC) and TB complete regimen (TBCR). Compared to individuals with TBC and with TBCR, individuals with TBFMA tended to die earlier and their mortality was significantly higher (HRTBFMA-TBC = 3.0, 95% confidence interval: 2.5-3.6, HRTBFMA-TBCR = 2.9, 95% CI: 2.5-3.4, P < 0.0001). Those who were naïve to antiretroviral therapy, were farmers, had lower CD4 counts (≤200 cells/µL) and were ≥ 50 years of age were at the highest risk of mortality. Mortality risk for participants with TBFMA was significantly higher across all stratifications except those with a CD4 count of ≤200 cells/µL. CONCLUSIONS: Earlier and long-term mortality among those with HIV/MTB co-infection is a significant problem when TB treatment fails or is inadequate.

[Analysis of clinical characteristics of severe and critically ill influenza A (H1N1)].

OBJECTIVE: To investigate the clinical features, laboratory results, chest CT imaging manifestations and treatments of severe and critical influenza A (H1N1), and to analyze the relationship with the prognosis. METHODS: The clinical data of 54 adult patients with severe and critical H1N1 admitted to the Fourth People's Hospital of Nanning from November 2018 to February 2019 were analyzed retrospectively. Throat swab specimens of the patients were determined for nucleic acid detection of influenza A (H1N1) virus, and all of the patients were confirmed. The gender, age, course of disease, underlying diseases, symptoms, body temperature, hospital stays, chest CT findings and laboratory results were collected, and the treatments and prognosis were recorded. RESULTS: Of 54 patients, 38 patients were enrolled in severe group, and 16 in critical group. Fever, cough, sputum, shortness of breath and so on could be found in the two groups. The CD4+T lymphocytes were less than normal reference value (410-1 590/µL) in both groups. The chest CT findings manifestations of severe group were scattered patchy shadows and ground glass appearance, all of them were cured and discharged after antiviral, antibiotics, and oxygen treatment. In critical group, the time in hospital was longer, the disease progresses varied faster, the shortness of breath was more apparent, and a large patch of fuzzy and real change shadows on both lungs could be found from CT findings. Compared with the severe group, creatine kinase (CK), lactic dehydrogenase (LDH), C-reactive protein (CRP) and procalcitonin (PCT) levels in the critical group were increased more significantly [CK (U/L): 704.50 (908.50) vs. 146.00 (220.75), LDH (U/L): 614.50 (492.25) vs. 217.00 (142.75), CRP (mg/L): 85.65 (56.13) vs. 18.80 (50.63), PCT (µg/L): 1.30 (5.00) vs. 0.10 (0.16), all P < 0.01], white blood cells count (WBC) and neutrophil ratio were also increased more significantly [WBC (×109/L): 12.37±7.63 vs. 8.29±3.32, neutrophil ratio: 0.81±0.11 vs. 0.75±0.11] without statistical differences (both P > 0.05). Nine patients in critical group were cured with cure rate of 56.25%. Seven patients died with mortality of 43.75%, including 2 patients with acquired immunodeficiency syndrome (AIDS) and uremia respectively, who had multiple organ failure (MOF) on admission and waive the mechanical ventilation treatment; 3 patients complicated with acute renal failure but abandon hemodialysis; 1 patient with nasopharyngeal carcinoma radiotherapy after operation; and 1 patient with chronic renal failure uremia period combined multiple drug-resistant bacteria infection, and died from MOF finally. CONCLUSIONS: The patients with severe and critical influenza A (H1N1) show fever, cough, dyspnea, and organ dysfunction in varying degrees. Severe patients were mainly pulmonary lesions, while critical patients show MOF such as heart, lung and kidney, and the lesions progressed rapidly. The major cause of death for critical influenza A (H1N1) may be chronic underlying diseases and MOF.