Persistent type I interferon (IFN-I) signaling contributes to immune exhaustion and promotes HIV-1 persistence. While we and others have demonstrated that blocking IFN-I signaling in vivo restores anti-HIV-1 T-cell function and reduces viral reservoirs, the underlying mechanism remains unclear. Here, we showed that in humanized mice (hu-mice) and cells from people living with HIV-1 (PLWH), IFN-I signaling impaired mitochondrial activity in CD8+ T cells during chronic HIV-1 infection with effective antiretroviral therapy. Reprogramming immunometabolism by transient inhibition of glycolysis with 2-deoxy-D-glucose (2-DG) rescued mitochondrial activity, reversed aberrant immune activation, and enhanced CD8+ T-cell activity in HIV-infected hosts, both ex vivo and in vivo. When combined with an HIV-1 reservoir-activating agent, 2-DG reduced the viral reservoir size in hu-mice and suppressed HIV-1 amplification in cells from PLWH. These findings indicate that 2-DG-mediated immunometabolic reprogramming represents a novel strategy to restore host immunity and control HIV-1 reservoirs.

Opportunistic infections (OIs) remain a major clinical challenge in people living with HIV/AIDS (PLWHA), and CD4⁺ T-cell counts alone may not fully reflect immune dysfunction. This study aimed to establish predictive cut-off values for lymphocyte subsets to improve the assessment of OIs risk in PLWHA. Peripheral blood samples were collected to assess lymphocyte subsets using flow cytometry between August 17, 2021, and April 19, 2023. Correlation analysis was performed to identify lymphocyte subset indicators strongly associated with CD4+ T-cell counts. Receiver operating characteristic (ROC) analysis was then performed to determine cut-off values for predicting OIs. Finally, the performance of cut-off values was tested in hospitalized PLWHA. Even when CD4 cell counts were above 200 cells/µl or close to normal, patients with abnormal immunological phenotypes had a higher prevalence of OIs. Other lymphocyte subsets have a high positive correlation with CD4+ T-cell counts, with the exception of CD3+CD8+/CD3+, CD3+HLA-DR+/CD3+, and CD3+CD8+HLA-DR+/CD8+, which are strongly negatively correlated. The correlation coefficients of CD4+CD28+ cells were particularly close to 1.0. Furthermore, CD4+CD28+ T lymphocytes demonstrated an AUC of 0.819 with a cut-off of 129 cells/µl, second only to CD4⁺ T cells in terms of AUC. Furthermore, the AUCs for CD3+CD4+CD45RA+, CD3+CD4+CD45RO+, CD3+CD4+CD25+CD127low, CD45RA+CD3+CD4+CD25+CD127low, and CD45RO+CD3+CD4+CD25+CD127low T cells were all > 0.800, with sensitivities exceeding 90

BackgroundDespite the success of antiretroviral therapy (ART) for human immunodeficiency virus (HIV) in China, immune non-response (INR) remains critical for the long-term quality of life of people living with HIV (PLWH). Although the consensus on diagnosis and management of immunological non-responders in HIV infection (Version 2023) was published in China to standardize diagnostic criteria, prediction models for INR based on these criteria remain scarce. This study aims to develop and validate a nomogram model for early identification of INR risk based on the diagnostic criteria in this consensus, so as to facilitate clinical intervention.MethodsIn this retrospective study, the primary cohort included 615 PLWH who initiated ART and completed over 4 years of follow-up at Zhongnan Hospital of Wuhan University (January 2016 to May 2025). They were randomly split into a training set (n=433) and an internal validation set (n=182) in a 7:3 ratio. An external validation set comprised 213 PLWH from Xishui County People’s Hospital (January 2012 to August 2025). Least absolute shrinkage and selection operator (LASSO) and multivariable logistic regression were used to identify independent predictors of INR, and a nomogram was constructed. The receiver operating characteristic (ROC) curves, calibration curves, and decision curve analysis (DCA) were employed to evaluate the discrimination, calibration, and clinical utility of the model in the training set and validation sets, respectively.ResultsLASSO regression identified seven candidate variables: age at ART initiation, baseline CD4+ T cell count, body mass index (BMI), white blood cell count (WBC), hemoglobin (Hb), aspartate aminotransferase (AST), and World Health Organization (WHO) clinical stage. Subsequent multivariable logistic regression confirmed baseline CD4+ T cell count (p < 0.001), age at ART initiation (p = 0.001), and AST level (p = 0.014) as independent INR predictors. The resulting nomogram demonstrated area under the curve (AUC) values of 0.896, 0.903, and 0.766 in the training, internal validation, and external validation sets, respectively. The calibration curve and DCA further indicated satisfactory consistency and clinical net benefit.ConclusionThe developed nomogram effectively predicts INR risk in PLWH initiating ART, providing clinicians a practical tool for individualized management to improve patient prognosis.

Strategies that concurrently reactivate latent reservoirs and enhance immune-mediated clearance hold significant promise for achieving an HIV cure. Here, we developed hyperIL-15×sCD4-Fc (15×sCD4-Fc), a fusion protein that integrates latency reactivation, targeted immune engagement, and effector-mediated killing into a single molecule. This agent not only potently reverses HIV-1 latency in CD4+ T cells from people living with HIV-1 (PLWHs) through coordinated IL-15 receptor signaling and sCD4-mediated HIV-1 envelope (Env) engagement but also enhances the antigen-specific CD8+ T cell response in peripheral blood mononuclear cells (PBMCs) derived from PLWHs. Furthermore, 15×sCD4-Fc enables Env-specific elimination of reactivated latently infected cells by NK cells while preventing off-target cytotoxicity. In PBMCs from PLWHs, 15×sCD4-Fc reduced replication-competent HIV-1 DNA by 93.8%. In antiretroviral-treated HIV-1-infected humanized mice, the molecule demonstrated both safety and efficacy in diminishing the viral reservoir in lymphoid organs. This spatiotemporally coupled approach to reservoir exposure and immune recognition establishes a clinically viable strategy for clearing the HIV-1 reservoir.

Objective The immune responses of COVID-19 convalescent patients have not been well described. Methods Blood from thirty COVID-19 convalescent patients who were virus-free were collected. Their clinical laboratory findings and SARS-CoV-2-specific humoral and cellular immunity were detected. Results At 283 days after diagnosis of SARS-CoV-2 infection, the levels of clinical laboratory indicators and lymphocyte subtypes returned to normal levels. However, the ratio of memory/naive CD4+ T lymphocytes cells was greater in COVID-19 convalescent patients and severe COVID-19 convalescent patients, when compared with that in healthy blood donors (P=0.0135) and non-severe patients (P=0.0431), respectively. The levels of anti-SARS-CoV-2-IgM (P=0.014), S1-IgM (P=0.0004) and RBD-IgM (P=0.0002) in severe COVID-19 patients were all significantly greater than those in non-severe COVID-19 patients. When the serums of COVID-19 convalescent patients were diluted as 1:125, the predictive of serum neutralization capabilities were persistent in all patients. SARS-CoV-2-specific T cells were generated and maintained in majority of tested convalescent COVID-19 patients, regardless of the severity of disease in acute phase. Conclusion At 283 days after diagnosis of SARS-CoV-2 infection, specific cellular and humoral immunity against SARS-CoV-2 could be detectable. The severity of disease in acute phase cannot affect the strength of cellular and humoral immunity in convalescent phase.

IntroductionThis study aimed to assess the diagnostic values of bronchoalveolar lavage fluid (BALF) real-time polymerase chain reaction (PCR) and BALF metagenomic next-generation sequencing (mNGS) for Pneumocystis jirovecii pneumonia (PJP) in patients infected with human immunodeficiency virus (HIV).MethodsA total of 99 HIV-infected PJP patients and 61 HIV-infected patients diagnosed with non-PJP pneumonia between March 2019 and December 2022 were enrolled. P. jirovecii and multiple other co-pathogens detected in BALF by mNGS were analyzed. The clinical final diagnosis was employed as a benchmark. We compared the diagnostic performance of mNGS in PJP with serum BDG and BALF real-time PCR. The mixed infections detected by mNGS and modifications of antimicrobial treatment were also analyzed.ResultsThe sensitivity of mNGS test of BALF samples reached 85.86%, which was significantly higher than serum BDG (39.39%, P < 0.001). The sensitivity of BALF P. jirovecii PCR (84.85%) was similar with mNGS (P > 0.05). The specificity of mNGS (100%) was also same as PCR (100.0%), and superior to serum BDG (88.52%, P < 0.001). Besides, mNGS performs remarkably well in identifying co-pathogens of PJP patients infected with HIV. In addition to P. jirovecii, 82 cases (82.83%) of other co-pathogens were identified based on mNGS. Moreover, thirty-four patients (34.34%) increased therapeutic dose of trimethoprim-sulfamethoxazole (TMP-SMZ) based on BALF P. jirovecii PCR. Based on the mNGS results, initial antimicrobial treatment was modified in 86.87% (86/99) of PJP patients.ConclusionBALF mNGS and real-time PCR are two powerful techniques for rapid diagnosis of PJP with high specificity and sensitivity. Moreover, the benefit of mNGS is that it may identify other organisms besides PJP and it may benefit proper and prompt treatment.

To evaluate clinical value of metagenomic next-generation sequencing (mNGS) in people living with HIV/AIDS (PLWHA) who had CNS disorders. Cerebrospinal fluid (CSF) samples from 48 PLWHA presenting with CNS disorders were sequenced using mNGS and compared with clinical conventional diagnostic methods. In total, 36/48 ss(75%) patients were diagnosed with pathogen(s) infection by mNGS, and the positive detection proportion by mNGS was higher than that by clinical conventional diagnostic methods (75% vs 52.1%, X 2 = 5.441, P = 0.020). Thirteen out of 48 patients (27.1%) were detected with 3–7 pathogens by mNGS. Moreover, 77 pathogen strains were detected, of which 94.8% (73/77) by mNGS and 37.0% (30/77) by clinical conventional methods ( X 2 = 54.206, P < 0.001). The sensitivity and specificity of pathogens detection by mNGS were 63.9% (23/36) and 66.7% (8/12), respectively, which were superior to that by clinical conventional methods (23/36 vs 9/25, X 2 = 4.601, P = 0.032; 8/12 vs 5/23, X 2 = 5.029, P = 0.009). The application of mNGS was superior for its ability to detect a variety of unknown pathogens and multiple pathogens infection, and relatively higher sensitivity and specificity in diagnosis of CNS disorders in PLWHA.

Objective:To evaluate the clinical value of metagenomic next-generation sequencing (mNGS) of cerebrospinal fluid in acquired immune deficiency syndrome (AIDS)? complicated with central nervous system (CNS) infection, and to investigate the pathogetic spectrum.Methods:AIDS patients complicated with CNS infection in Zhongnan Hospital of Wuhan University from January 2021 to August 2022 were analyzed, including the clinical characteristics and pathogen information detected by mNGS and routine etiological examination. The pathogens detected by routine etiological examination and mNGS were compared, and the consistency with clinical diagnosis was evaluated.Results:Among the 61 patients with CNS infection, 60 patients (98.4%, 60/61) were identified by mNGS combined with routine etiological examination, which was significantly higher than that of routine etiological examination alone (49.2%, 30/61) and that of mNGS alone (83.6%, 51/61), with significant differences (χ2 = 38.125, P < 0.001; χ2 = 8.093, P = 0.004). The positive rate of mNGS 83.6% (51/61) was significantly higher than that of routine etiological examination [49.2% (30/61)], with significant difference (χ2 = 16.201, P < 0.001). The detection rate of two or more pathogens by mNGS was 41.0% (25/61), which was significantly higher than that by routine etiological examination (9.8%, 6/61), with significant difference (χ2 = 15.612, P < 0.001). For the detection rate of cryptococcal meningitis, conventional detection method [14.8% (9/61)] was in good agreement with mNGS [13.1% (8/61)], and the consistency with clinical diagnosis were 90% (9/10) and 80% (8/10), respectively. At least one virus was detected in 43 (70.5%) patients, of which 24 (39.3%) cases detected human herpesvirus 4 (EBV), 15 (24.6%) cases detected human herpesvirus 5 (CMV), and 8 (13.1%) cases detected JC polyomavirus. Total of 18 strains of fungi were detected in 14 (23.0%) patients, including 8 strains of Cryptococcus neoformans, 4 strains of Aspergillus, 3 strains of Candida parapsilosis, 1 strain of Talaromyces marneffei, 1 strain of Pneumocystosis jirovecii and 1 strain of Dosporium apiospermum. There were 8 (13.1%) atypical pathogens detected, including 3 strains of Toxoplasma gondii, 3 strain of Treponema pallidum, 1 strain of Legionella and 1 strain of Rickettsia felis. Tuberculosis and non-tuberculous mycobacterium were detected in 2 (6.6%) cases respectively. Total of 3 cases died, the mortality rate was 4.9% (3/61).Conclusions:Mixed infections are common in advanced AIDS patients complicated with CNS infection, and the pathogetic spectrum includes virus, fungi, bacterial and atypical pathogens. mNGS combined with routine etiological examination could significantly improve the rapid pathogen detection rate of intracranial infections and the survival rate of patients.

Objective:To investigate the changing characteristics of the immunocytes after human immunodeficiency virus (HIV) infection and the dynamic changing patterns of different disease stages.Methods:Lymphocyte subsets of 173 patients with HIV/acquired immunodeficiency syndrome (AIDS) hospitalized in Department of Infectious Diseases, Zhongnan Hospital of Wuhan University from August 17th, 2021 to September 14th, 2022 (research group) were analyzed by flow cytometry, and were compared with 1 086 healthy individuals (control group). AIDS was staged according to the CD4+ T lymphocyte count; The lymphocyte subset classification counts of patients with HIV/AIDS at different AIDS stages were compared. The absolute counts of different types of lymphocytes in the measurement data were normally distributed, expressed by ±s, and the mean values of all types of lymphocytes between patients with HIV/AIDS and control group were compared by two independent samples t-tests.Results:The absolute counts of T lymphocytes, B lymphocytes and NK cells of patient in research group were (907 ± 105) cells/μl, (128 ± 25) cells/μl and (176 ± 16) cells/μl, respectively, which were significantly lower than those in control group (t = 8.508, P < 0.001; t = 8.265, P < 0.001; t = 18.552, P < 0.001). CD4+CD28+ cells and CD8+CD28+ cells of patient in research group were (123 ± 25) cells/μl and (245 ± 98) cells/μl, which were both significantly lower than those of control group (t = 28.522, P < 0.001; t = 5.820, P < 0.001). CD3+CD8+HLA-DR+ cell counts and percentages of patient of research group were (476 ± 129) cells/μl and 73.68%, respectively, which were both higher than those of control group (t = 8.482, P < 0.001; t = 34.651, P < 0.001). The absolute counts of na?ve and memory CD4+ T cells of patient of research group were (76 ± 9) cells/μl and (152 ± 12) cells/μl, respectively, which were significantly lower than those of control group (t = 19.823, P < 0.001; t = 20.815, P < 0.001). As the disease progressed, the absolute counts of CD4+CD28+ cells and CD8+CD28+cells gradually decreased from (540.77 ± 165.54) cells/μl and (452.57 ± 135.65) cells/μl to (13.56 ± 33.63) cells/μl and (102.96 ± 30.47) cells/μl, and the percentages of CD3+CD8+ HLA-DR+ cells increased from (36.00 ± 17.79)% to (58.29 ± 13.27)% of patients in research group.Conclusions:Abnormalities in lymphocyte count and function, as well as abnormal activation of the immune system are common in patients with HIV/AIDS. In late stage of AIDS, impaired lymphocyte function and abnormal immune activation are more significant.

Long noncoding RNAs (lncRNAs) play important roles in regulating HIV-1 infection and virus-host interactions. However, it is unclear whether and how ART alters lncRNAs in HIV-infected patients. In the present study, we investigated changes of lncRNAs in PBMCs from HIV-1 patients pre- and post-ART. We identified a total of 974 lncRNAs whose expression was restored to normal levels after ART. Cis-acting analysis showed that six lncRNAs have cis-regulated target genes, among which RP11-290F5.1 and interferon regulatory factor 2 (IRF2) were reported to promote HIV replication. Furthermore, we found that lncRNA CTB-119C2.1, which regulates most mRNAs with differential expression in PBMCs from HIV-1 infected patients after ART, was significantly upregulated by RNA-seq and qRT-PCR assays. KEGG analysis of CTB-119C2.1-associated genes revealed that most of the genes are involved in the p53 signaling pathway and pathways related to cell cycle and DNA replication. Our findings thus reveal the dynamic change of lncRNAs in people living with HIV-1 pre- and post-ART and warrant further investigation of the role of lncRNAs in HIV-1 pathogenesis and treatment.

Background:Severe fever with thrombocytopenia syndrome (SFTS) is an emerging zoonosis caused by a novel bunyavirus. Until recently, the SFTS related acute kidney injury (AKI) was largely unexplored. This study aimed to investigate the clinical characteristics and outcomes of AKI in patients with SFTS.Methods:The non-AKI and AKI groups were compared in terms of general characteristics, clinical features, laboratory parameters and cumulative survival rate. The independent risk factors for in-hospital mortality in patients with SFTS were analyzed by multivariate logistic regression to identify the population with poor prognosis.Results:A total of 208 consecutive patients diagnosed with SFTS were enrolled, including 153 (73.6%) patients in the non-AKI group and 55 (26.4%) patients in the AKI group. Compared with patients without AKI, patients with AKI were older and had a higher frequency of diabetes. Among these laboratory parameters, platelet count, albumin and fibrinogen levels of patients with AKI were identified to be significantly lower than those of patients without AKI, while ALT, AST, ALP, triglyceride, LDH, BUN, uric acid, creatine, Cys-C, β2-MG, potassium, AMY, lipase, CK-MB, TnI, BNP, APTT, thrombin time, D-dimer, CRP, IL-6, PCT and ESR levels were significantly higher in patients with AKI. A higher SFTS viral load was also detected in the AKI patients than in the non-AKI patients. The cumulative survival rates of patients at AKI stage 2 or 3 were significantly lower than those of patients without AKI or at AKI stage 1. However, there was no significant difference in the cumulative survival rates between patients without AKI and those with stage 1 AKI. Univariate and multivariate binary logistic regression analyses demonstrated that stage 2 or 3 AKI was an independent risk factor for in-hospital mortality in patients with SFTS.Conclusion:AKI is associated with poor outcomes in patients with SFTS, especially patients at AKI stage 2 or 3, who generally have high mortality. Our findings support the importance of early identification and timely treatment of AKI in patients with SFTS.

Background:To analyze the changing characteristics of continuous monitoring of refined lymphocyte subsets in people living with HIV/AIDS (PLWHA) during ART period.Methods:Refined lymphocyte subsets was continuously monitored using flow cytometry for 173 PLWHA, who were hospitalized in Zhongnan Hospital of Wuhan University from August 17, 2021 to September 14, 2022. The effect of ART status and duration of ART on changes of refined lymphocyte subsets were compared in different groups. Then, the levels of refined lymphocyte subsets in PLWHA treated for more than 10 years were compared to those of 1086 healthy individuals.Results:In addition to conventional CD4+ T lymphocytes and CD4+/CD8+ ratio, gradually increasing in numbers of CD3+CD4+CD45RO cells, CD3+CD4+CD45RA cells, CD45RA+CD3+CD4+CD25+CD127low and CD45RO+CD3+CD4+CD25+CD127low cells were found with the increase of ART duration. The number of CD4+CD28+ cells and CD8+CD28+ cells were 174/ul and 233/ul at 6 months post-ART, which gradually increased to 616/ul and 461/ul after ART initiation more than 10 years. Moreover, in ART ≤ 6 months, 6 months-3years, 3-10 years and >10 years groups, the percentage of CD3+CD8+HLA-DR+/CD8 were 79.66%, 69.73%, 60.19% and 57.90%, respectively, and the differences between groups showed statistical significance (F=5.727, P=0.001). For those PLWHA with ART more than 10 years, the levels of CD4+ T lymphocytes, CD3+CD4+CD45RO cells, CD3+CD4+CD45RA cells, CD4+CD28+ cells and CD8+CD28+ cells can increase to levels similar to those of healthy control. However, for those PLWHA with ART more than 10 years, CD4+/CD8+ ratio was 0.86 ± 0.47, which was lower than that of healthy control (0.86 ± 0.47 vs 1.32 ± 0.59, t=3.611, P=0.003); absolute counts and percentage of CD3+CD8+HLA-DR+ cells were 547/ul and 57.90%, which were higher than those of healthy control(547/ul vs 135/ul, t=3.612, P=0.003; 57.90% vs 22.38%, t=6.959, P<0.001).Conclusion:Persistent ART can gradually improve the immune status of PLWHA, which is manifested in the increase of lymphocytes, function recovery of lymphocytes and reduction of aberrant activation status of the immune system. After 10 years of standardized ART, most lymphocytes could return to levels of healthy persons, although it may take longer to complete recovery for CD4+/CD8+ ratio and CD3+CD8+HLA-DR+ cells.

MicroRNAs (miRs) are 18-25 nucleotides non-coding RNAs, which contribute to tumorigenesis. Previous studies have demonstrated that miR-199a-3p is dysregulated in human nasopharyngeal carcinoma (NPC), but its role in NPC progression still largely unknown. The current study aimed to determine the potential role of miR-199a-3p in NPC progression and the underlying mechanisms. In this study, miR-199a-3p was found to be prominently down-regulated in NPC tissues and cells. The cellular assay showed that transfection of miR-199a-3p markedly repressed the migration, invasion and induced epithelial-mesenchymal transition (EMT) in both 5-8F and CNE-2 cell lines. By dual-luciferase reporter, western blotting and gas chromatography assays, we found that SCD1 is not only highly expressed in NPC tissues and negatively associated with the prognosis of NPC patients but also can be apparently downregulated by miR-199a-3p in NPC cells, suggesting that SCD1 is a direct target gene of miR-199a-3p. Moreover, inhibition of miR-199a-3p expression activated PI3K/Akt signaling and up-regulated the expression of MMP-2. With tumor xenograft models in nude mice, we also showed that miR-199a-3p repressed tumor growth in vivo. Our study demonstrated that miR-199a-3p inhibited migration and invasion of NPC cells through downregulating SCD1 expression, thus providing a potential target for the treatment of NPC.

Background and objectives Although fish consumption or omega-3 intake is associated with cardio- cerebrovascular disease including stroke, their correlation is still controversial. Therefore, this meta-analysis is to identify the relationship between the risk of stroke and fish consumption or omega-3 intake. Methods and study design We searched the PubMed, EMBASE and Cochrane Library databases as of May 2019. Multivariateadjusted risk ratios (RRs) with 95% confidence interval (CI) for stroke in different level intake of fish or Longchain omega-3 polyunsaturated fatty acids (LC ω3-PUFAs) were pooled using a random-effects meta-analysis. A dose-response analysis was conducted with the 2-stage generalized least-squares trend program. Results Our meta-analysis identified a total of 17 prospective cohort studies including 14986 strokes events in 672711 individuals. Meta-analysis revealed that the higher fish consumption was significantly associated with lower risk of stroke (RR=0.871, 95% CI: 0.779-0.975, p=0.016), especially with ischemic stroke (RR=0.808, 95% CI: 0.696- 0.937, p=0.005). Meantime, the combined RR of total stroke was 0.859 (95% CI: 0.769-0.959, p=0.007) for the highest versus lowest intake of LC ω3-PUFAs, and stratification analysis showed that higher LC ω3-PUFAs intake was associated with reduced stroke risk in women (RR=0.793, 95% CI: 0.706-0.891, p=0.000) but not in men. In addition, the dose-response analysis showed fish consumption with 1000g per month and LC ω3-PUFAs intake with 0.5g per month was associated with 17.3% (RR=0.927, 95% CI: 0.83-0.98) and 14% (RR=0.86, 95% CI: 0.78-0.95) lower risk of stroke, respectively. Conclusions Both fish consumption and LC ω3-PUFAs intake were negatively associated with the risk of stroke, especially in women, which suggest that increased intake of fishery products and LC ω3-PUFAs may benefit primary prevention of stroke.

Comorbidities are important for the disease outcome of COVID‐19, however, which underlying diseases that contribute the most to aggravate the conditions of COVID‐19 patients are still unclear. Viral clearance is the most important laboratory test for defining the recovery of COVID‐19 infections. To better understand which underlying diseases that are risk factors for delaying the viral clearance, we retrospectively analyzed 161 COVID‐19 clinical cases in the Zhongnan Hospital of Wuhan University, Wuhan, China between January 5 and March 13, 2020. The demographic, clinical and laboratory data, as well as patient treatment records were collected. Univariable and multivariable analysis were performed to explore the association between delayed viral clearance and other factors by using logistic regression. Survival analyses by Kaplan‐Meier and Cox regression modeling were employed to identify factors negatively influencing the viral clearance negatively. We found that hypertension and intravenous immunoglobulin adversely affected the time of viral RNA shedding. Hypertension was the most important risk factor to delay the SARS‐CoV‐2 virus clearance, however, the use of Angiotensin‐Converting Enzyme Inhibitors(ACEI)/Angiotensin Receptor Blockers(ARB) did not shorten the time for virus clearance in these hypertensive patients’ virus clearance. We conclude that patients having hypertension and intravenous immunoglobulin may delay the viral clearance in COVID‐19 patients.

Background: Many studies have shown that long noncoding RNAs (lncRNAs) derived from the host and human immunodeficiency virus (HIV) itself play important roles in virus-host interactions and viral pathogenesis. To identify potential key lncRNAs in the regulation of HIV pathogenesis, transcriptome analysis of peripheral blood mononuclear cells (PBMCs), which were derived from 6 HIV/acquired immunodeficiency syndrome (AIDS) subjects pre-HAART and post-HAART with effective control of plasma viremia (<20 HIV RNA copies/ml) and 6 healthy subjects, was performed by RNA sequencing (RNA-seq). Results: We identified a total of 974 lncRNAs whose expression levels were restored to normal after ART therapy. The results of the cis-acting analysis showed that only six lncRNAs have cis-regulated target genes, among which the target gene RP11-290F5.1, interferon regulatory factors 2 (IRF2), could promote HIV replication. We also identified lncRNA CTB-119C2.1, which regulates most mRNAs with differential expression between pre- and post-HAART, and the differences were significant. We selected lncRNA CTB-119C2.1 for qRT–PCR verification, and the results were consistent with those of RNA-seq. RAB3A and GADD45A, two of the lncRNA CTB-119C2.1-associated genes, have been shown to be associated with HIV infection. KEGG analysis of lncRNA CTB-119C2.1-associated genes revealed that most of the genes are involved in the p53 signaling pathway or pathways related to cell circulation and DNA replication Conclusion: In this study, we used RNA-seq to systematically compare the expression profiles of lncRNAs in HIV subjects between untreated and treated time points. We successfully identified some lncRNAs with differential expression during certain periods (no HIV infection, HIV infection before treatment, and after treatment). Their expression is associated with viral loads, and some of their regulating genes were found to be involved in HIV pathogenesis through bioinformatic analysis. These findings could help to reveal the underlying molecular mechanism of the progression of AIDS.

Objective:To analyze the characteristics of clinical and laboratory data of coronavirus disease 2019 (COVID-19) patients with different severity on admission, and to find a laboratory indexes for early warning mild/moderate development into severe/critical type.Methods:Total of 44 patients confirmed with COVID-19 were included in the Department of Infectious Diseases, Zhongnan Hospital of Wuhan University in February 2020, retrospectively. Patients with COVID-19 were divided into mild/moderate group (28 patients) and severe/critical group (16 patients). Blood routine tests [white blood cell (WBC), neutrophil (NEU), lymphocyte (LYM), monocytes (MON), platelet (PLT)], biochemical examinations [alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TBil), albumin (ALB), creatinine (CREA)], inflammatory factors tests [C-reactive protein (CRP), interleukin-6 (IL-6)], new coronavirus nucleic acid tests and peripheral blood lymphocyte subsets tests performed within 3 days on admission were collected and analyzed by t test, rank sum test or chi-square test, respectively. The risk factors of severe disease were analyzed by Binary Logistic regression analysis.Results:Levels of peripheral blood WBC, NEU, MON, PLT, LYM, and absolute count of CD3+ T lymphocytes, CD3+CD4+T lymphocyte, CD3+CD8+T lymphocytes, CD4+/CD8+ T, CD19+B lymphocytes, CD16+CD56+NK cells of patients in severe/critical group were all lower than those of mild/moderate group, among which, the absolute count of CD3+ T lymphocytes (t = 2.24, P = 0.03), CD3+CD4+ T lymphocytes (t = 2.148, P = 0.037) and peripheral blood lymphocytes (t = 2.039, P = 0.047) were decreased with significant difference. Serum IL-6 level was 21.06 (11.02, 36.43) pg/ml of patients in severe/critical group and 6.13 (3.14, 12.54) pg/ml in mild/moderate group, with significant difference (Z = 2.952, P = 0.003). Novel coronavirus nucleic acid Ct values ??of respiratory tract specimens and fecal specimens of patients in severe/critical group were (30.03 ± 1.196) and (33.12 ± 1.48), which were significantly lower than those of mild/moderate group: (33.56 ± 0.75) of respiratory tract specimens (t =2.634, P = 0.012) and (35.76 ± 0.98) of fecal specimens (t =1.545, P = 0.130) with significant differences. The length of hospital stay of patients in severe/critical group was (25.06 ± 3.01) days, which was significantly higher than that of mild/moderate group [(11.79 ± 1.06) days], with significant difference (t = 4.986, P < 0.001). Binary Logistic regression analysis found that IL-6 (OR = 1.059, 0.039) and the length of hospital stay (OR = 1.173, 0.005) were both the influencing factors of the severity of patients with COVID-19.Conclusions:There were significant differences in lymphocyte subsets, serum IL-6 level and viral nucleic acid levels between COVID-19 patients with varying severity, and the length of hospital stay and abnormally increased IL-6 were risk factors for severe disease, and both were of great significance in predicting the severity and clinical classification of COVID-19 patients at early stage.

长链非编码RNA(long noncoding RNA, lncRNA)在多种生物通路中具有重要的调控功能，在病毒复制和先天性免疫应答调控网络中发挥着重要作用。本研究讨论了与人类免疫缺陷病毒(human immunodeficiency virus，HIV)感染相关的复杂的lncRNA网络，来自人类宿主和HIV自身的lncRNA影响了HIV复制、潜伏期的建立及维持、储存库的清除、病毒的重新激活等过程。由于有效的HIV疫苗或治愈方法的缺乏，以及目前大流行的规模，深入了解lncRNA和HIV之间的复杂调控网络将有助于制订治疗HIV感染的新战略。

The humoral and cellular immunity of convalescent COVID-19 patients is involved in pathogenesis and vaccine immunity. In this study, through CoV-psV neutralization assay and IFN-γ ELISpot testing in 30 cases of COVID-19 patients after 9 months post-SARS-CoV-2 infection, it found that the ratio of memory/naive CD4+ T lymphocytes cells and levels of anti-SARS-CoV-2-IgM and RBD-IgM were slightly but significantly higher in COVID-19 severe convalescent patients than that in non-severe patients. The specific cellular and humoral immunity against SARS-CoV-2 were detectable, regardless of the severity of the disease in the acute phase. This information may help understanding the immune status after SARS-CoV-2 infection.

Different primers/probes sets have been developed all over the world for the nucleic acid detection of SARS-CoV-2 by quantitative real time polymerase chain reaction (qRT-PCR) as a standard method. In our recent study, we explored the feasibility of droplet digital PCR (ddPCR) for clinical SARS-CoV-2 nucleic acid detection compared with qRT-PCR using the same primer/probe sets issued by Chinese Center for Disease Control and Prevention (CDC) targeting viral ORF1ab or N gene, which showed that ddPCR could largely minimize the false negatives reports resulted by qRT-PCR [Suo T, Liu X, Feng J, et al. ddPCR: a more sensitive and accurate tool for SARS-CoV-2 detection in low viral load specimens. medRxiv [Internet]. 2020;2020.02.29.20029439. Available from: https://medrxiv.org/content/early/2020/03/06/2020.02.29.20029439.abstract]. Here, we further stringently compared the performance of qRT-PCR and ddPCR for 8 primer/probe sets with the same clinical samples and conditions. Results showed that none of 8 primer/probe sets used in qRT-PCR could significantly distinguish true negatives and positives with low viral load (10(-4) dilution). Moreover, false positive reports of qRT-PCR with UCDC-N1, N2 and CCDC-N primers/probes sets were observed. In contrast, ddPCR showed significantly better performance in general for low viral load samples compared to qRT-PCR. Remarkably, the background readouts of ddPCR are relatively lower, which could efficiently reduce the production of false positive reports.