Transfer of Invitro CD4 + T Cells with Hypomethylation of Perforin Promoter into Rats' Abdomens Causes Autoimmune Emphysema.

Our previous study suggested that hypomethylation of perforin promoter of CD4 + T cells might be involved in the pathogenesis of autoimmune emphysema of rats. Whether transfer of this kind of cells hypomethylated in vitro into naive immunocompetent rats also results in emphysema is unknown yet. To test the hypothesis above, thirty Sprague Dawley (SD) rats were randomly divided into three groups: a model group (n = 10), a normal control group (n = 10) and a sham operation group (n = 10). In the model group, spleen-derived CD4 + T cells of normal rats were treated with 5-azacytidine (5-Aza), complete Freund's adjuvant and Phosphate Buffered Saline (PBS), then transferred into naive immunocompetent rats. The normal control group was injected with CD4 + T lymphocytes from spleens of normal rats and the same amount of adjuvant and PBS as above. In sham operation group, normal rats were injected intraperitoneally with complete Freund's adjuvant and PBS. Histopathological evaluations (mean linear Intercept (MLI) and mean alveolar numbers (MAN)), anti-endothelial cell antibodies (AECA) in serum and bronchoalveolar lavage fluid (BALF), lung vascular endothelial growth factor (VEGF)), the apoptotic index (AI) of alveolar septal cells and the methylation levels of perforin promoter of CD4 + T cells were investigated. The levels of the methylation above and MAN were lower in the model group than in the control and the sham operation group, while the AECA in serum and BALF, VEGF, MLI and the AI were greater (all p < 0.05). The methylation levels of perforin promoter were positively correlated with the MAN (r = 0.747, p < 0.05) and negatively correlated with AI, AECA, MLI, and VEGF (r was -0.789, -0.746, -0.743, -0.660, respectively, all p < 0.05). This study suggests that transfer of invitro CD4 + T cells with hypomethylation of perforin promoter into rats causes autoimmune emphysema, possibly by increasing expression of VEGF and promoting alveolar septal cell apoptosis.

lncRNA NEAT1 aggravates sepsis-induced lung injury by regulating the miR-27a/PTEN axis.

Sepsis is an acute inflammatory reaction and a cause of acute respiratory distress syndrome (ARDS). In the present study, we explored the roles and underlying mechanism of the lncRNA Nuclear enriched abundant transcript 1 (NEAT1) in ARDS. The expression levels of genes, proteins and pro-inflammatory cytokines in patients with ARDS, LPS-stimulated cells and septic mouse models were quantified using qPCR, western blotting and ELISA assays, respectively. The molecular targeting relationship was validated by conducting a dual-luciferase reporter assay. Cell proliferation was assessed using the Cell Counting Kit-8 (CCK-8) assay. The cell cycle phase was determined by flow cytometry assay. The expression levels of NEAT1 and pro-inflammatory cytokines were higher in patients with ARDS and septic models than in controls. Knockdown of NEAT1 significantly increased cell proliferation and cycle progression and prolonged mouse survival in vitro and in vivo. Mechanistically, miR-27a was identified as a downstream target of NEAT1 and directly inhibited PTEN expression. Further rescue experiments revealed that inhibition of miR-27a impeded the promoting effects of NEAT1 silence on cell proliferation and cycle progression, whereas inhibition of PTEN markedly weakened the inhibitory effects of NEAT1 overexpression on cell proliferation and cycle progression. Altogether, our study revealed that NEAT1 plays a promoting role in the progression of ARDS via the NEAT1/miR-27a/PTEN regulatory network, providing new insight into the pathologic mechanism behind ARDS.

Clinical option of pemetrexed-based versus paclitaxel-based first-line chemotherapeutic regimens in combination with bevacizumab for advanced non-squamous non-small-cell lung cancer and optimal maintenance therapy: evidence from a meta-analysis of randomized control trials.

BACKGROUND: In the era of immunotherapy, it is still unclear which is the best first-line therapy for patients with oncogenic driver negative advanced non-squamous non-small cell lung cancer (NS-NSCLC) who cannot tolerate immunotherapy, or subsequent therapy for patients with oncogenic driver positive NS-NSCLC whose disease progressed on prior targeted therapy. To assess the optimal choice of first-line and maintenance treatment regimens, we performed a meta-analysis of prospective randomized controlled clinical trials (RCTs) of patients with NS-NSCLC on bevacizumab combined with chemotherapy. METHODS: All eligible RCTs comparing pemetrexed-platinum with or without bevacizumab (PP ± B) and paclitaxel-carboplatin with bevacizumab (PC + B) as a first-line therapy, or comparing bevacizumab plus pemetrexed (Pem + B) and bevacizumab alone (B) as a maintenance treatment for advanced NS-NSCLC, were included after systematically searching web databases and meeting abstracts. The main research endpoints were comparisons of overall survival (OS) and progression-free survival (PFS). The other endpoints were objective response rate (ORR), 1-year PFS rate (PFSR1y) and major grade 3/4 treatment-related adverse events. RESULTS: Data of 3139 patients from six RCTs were incorporated into analyses. Three RCTs were included in an analysis that compared PP ± B and PC + B as a first-line therapy for advanced NS-NSCLC. Patients treated with first-line PP ± B showed similar OS and ORR, but significantly improved PFS (hazard ratio [HR], 0.88) and PFSR1y (risk ratio [RR], 0.83), as compared to patients treated with PC + B (all P < 0.05). PP ± B resulted in higher rates of grade 3/4 anemia and thrombocytopenia, but lower rates of neutropenia, febrile neutropenia, and sensory neuropathy than PC + B (all P < 0.001). The other three RCTs were included in an analysis that compared Pem + B and B as a maintenance treatment. Compared with B, Pem + B maintenance treatment resulted in significant improvements in OS (HR, 0.88), PFS (HR, 0.64), and PFSR1y (RR, 0.70), but higher rates of anemia, thrombocytopenia, and neutropenia (all P < 0.001). CONCLUSION: Although the first-line PP + B regimen had longer PFS and PFSR1y than the PC + B regimen, no OS difference was observed. Addition of pemetrexed to bevacizumab as maintenance therapy significantly improved OS compared with bevacizumab maintenance alone, but led to more toxicity.

Immunotherapeutic effects of allogeneic mesenchymal stem cells on systemic lupus erythematosus.

Mesenchymal stem cells have been applied to treat graft versus host disease as they have immunosuppressive ability and can overcome the major histocompatibility complex-histocompatibility barrier. The potential of allogeneic mesenchymal stem cells in treating systemic lupus erythematosus (SLE) was investigated in this study. MRL/lpr mice which can develop acquired SLE-like phenotypes were selected as an animal model. Mesenchymal stem cells obtained from green fluorescent protein-transgenic ICR mice were infused into MRL/lpr mice at either the early or late stage of disease. The dosage was 1 × 106/mice per infusion. Mice were stratified into six groups including negative controls and those receiving one, two, three, four or five doses at 2-weekly intervals. The phenotypes were monitored regularly. After treatment, the spleen CD3+CD4-CD8- T and CD19+ B cells of two-dose mesenchymal stem cell-treated mice were significantly lower than those of the phosphate-buffered saline control. In terms of reducing the severity of SLE such as hair loss, skin ulcers, proteinuria and anti-dsDNA level, mesenchymal stem cells given at the early stage responded better and mice receiving two doses of mesenchymal stem cells performed better than those receiving either a lower dose (one dose) or higher doses (three, four or five doses). In conclusion, early treatment and an optimal dose of mesenchymal stem cells can effectively suppress the murine SLE model.

Hepatitis E virus seroprevalence among farmers, veterinarians and control subjects in Jilin province, Shandong province and Inner Mongolia Autonomous Region, China.

China is commonly considered to be a HEV-endemic region but limited epidemiological data for HEV among farmers and veterinarians are available. Thus, a case-control study was carried out to detect the seroprevalence and assess potential risk factors associated with the acquisition of HEV infection by farmers and veterinarians in China from July 2013 to May 2015. Three hundred veterinarians and 600 farmers recruited from Jilin province, Shandong province, and Inner Mongolia Autonomous Region and 600 control subjects matched by gender, age, and residence were detected for the presence of anti-HEV IgG and IgM antibodies using enzyme immunoassays. The seroprevalences of HEV infection in farmers, veterinarians, and control subjects were 34.8%, 26.7%, and 20.2%, respectively. Farmers (P < 0.001) and veterinarians (P = 0.027) have significantly higher seroprevalence than control subjects. The highest seroprevalence of HEV infection was detected in swine farmers (49.1%) and the lowest seroprevalence was found in cattle farmers (26.5%). In veterinarians, farm animal veterinarians have a higher seroprevalence than pet veterinarians, but the difference was not significant (P > 0.05). Residence area, contact with swine and exposure with soil were significantly associated with HEV infection in the study farmers; contact with swine and source of drinking water were significantly associated with HEV infection in the study veterinarians. These results implied the high prevalence of HEV and the considerable potential for the dissemination of HEV infection in farmers and veterinarians in China. J. Med. Virol. 89:872-877, 2017. © 2016 Wiley Periodicals, Inc.

Seroprevalence of hepatitis E virus among pregnant women and control subjects in China.

Hepatitis E infection, caused by the hepatitis E virus (HEV), is an important global public health concern, with particularly high mortality in pregnant women. China is generally judged to be an HEV-endemic area, but epidemiological data for HEV among pregnant women are limited. Between June 2011 and July 2013, a case-control study was conducted to estimate the seroprevalence and potential risk factors associated with the acquisition of HEV infection by pregnant women in China. Nine-hundred and ninety pregnant women who visited hospitals for antenatal follow-up or medication in Qingdao and Weihai and 965 control subjects matched by age, gender and residence were examined for the presence of anti-HEV IgG and IgM antibodies by enzyme immunoassays. Socio-demographic and behavioral characteristics from the study subjects were obtained. The overall prevalence of anti-HEV IgG in all 1,955 samples was 20.7%. In pregnant women, 16.2% of samples were anti-HEV IgG positive whereas, in control subjects 25.3% of samples were anti-HEV IgG positive, (P < 0.01). For anti-HEV IgM detection, 62 (3.2%) of the 1,955 serum samples were positive and the seroprevalence in pregnant women and control subjects was 2.6% and 3.6%, respectively. Age, contact with cats, contact with pigs and exposure to soil were found to be associated with HEV infection. These findings demonstrated the high prevalence of HEV and the considerable potential for the transmission of HEV infection in pregnant women in China.

Hypomethylation of perforin regulatory elements in CD4+ T cells from rat spleens contributes to the development of autoimmune emphysema.

BACKGROUND AND OBJECTIVE: It is widely accepted that perforin regulatory elements are hypomethylated in CD4+ T cells from patients with active lupus, but whether this is the case in autoimmune emphysema is not known. METHODS: Twenty rats were randomly divided into a normal control group and an emphysema group. Rat models of emphysema were established by intraperitoneal injection with xenogeneic endothelial cells. The levels of tumour necrosis factor-α, interleukin-8 and matrix metalloproteinase (MMP)-9 in bronchoalveolar lavage fluid (BALF) were measured, lung mean linear intercept and destructive index measured. Mean methylation of perforin gene promoter in CD4+ T cells and the expression of perforin were investigated. Terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling methods were used to examine the percentage of apoptotic cells in the alveolar septa. RESULTS: The levels of MMP-9 in BALF were higher in emphysema group than in control group (P < 0.05). The mean linear intercept and destructive index were higher in emphysema group than in control group (P < 0.05). The mean perforin gene promotor methylation of emphysema group was significantly decreased as compared with control group, while the expression levels of perforin gene were relatively higher (P < 0.05). There were more terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling-positive cells in the alveolar septa in control group than in emphysema group. CONCLUSIONS: Hypomethylation of perforin regulatory elements in CD4+ T cells may result in the lung septal cell apoptosis associated with the development of experimental autoimmune emphysema. MMP-9 may play an important role in the pathogenesis of this kind of disease.

[The effect of continuous renal replacement therapy on the outcome of severe pneumonia in patients receiving long-term immunosuppressants].

OBJECTIVE: To investigate the effect of continuous renal replacement therapy (CRRT) on the outcome of severe pneumonia in patients receiving long-term immunosuppressants. METHODS: Thirty-four patients, who had been treated with long-term immunosuppressants, were admitted for severe pneumonia. After admission, the dose of immunosuppressants including glucocorticoids was decreased, and the patients were divided into 2 groups: antibiotic treatment group (n = 16) and antibiotic + CRRT treatment group (n = 18). Before and after treatment, the changes of the patients' condition, chest CT and blood gas analysis were monitored. Biomarkers including C-reactive protein (CRP), interferon-γ (IFN-γ), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), interleukin-8(IL-8), white blood cell and neutrophil counts were determined. RESULTS: After treatment, the levels of CRP[(109 ± 24) vs (40 ± 13) mg/L], IFN-γ [(151 ± 28) vs (42 ± 12) ng/L], TNF-α [(301 ± 45) vs (118 ± 28) pg/L], IL-6 [(214 ± 45) vs (76 ± 23) pg/L], IL-8[(590 ± 121) vs (159 ± 60) pg/L], white blood cell count [(14.3 ± 5.7)×109/L vs (8.5 ± 2.7)×109/L], and neutrophil percentage [(91.3 ± 3.1)% vs (75.3 ± 2.6)%] decreased significantly in the antibiotic + CRRT group (P < 0.05) as compared to the antibiotic treatment group. In the antibiotic + CRRT group, blood gas showed significant improvement in pH [(7.30 ± 0.12) to (7.37 ± 0.18)], SaO2 [(80.6 ± 7.6)% to (91.9 ± 7.3)%] and PaO2 (41 ± 6) mm Hg (1 mm Hg = 0.133 kPa) to (71 ± 9) mm Hg. The patients' condition and chest CT abnormalities also improved more rapidly in the antibiotic + CRRT group. The 6-month survival was increased by 12.9% in the antibiotic + CRRT group as compared to the antibiotic group (P < 0.05). CONCLUSION: CRRT is effective in clearance of inflammatory mediators and may increase survival of severe pneumonia patients receiving long-term treatment of immunosuppressants.

SAM protects against alveolar septal cell apoptosis in autoimmune emphysema rats.

BACKGROUND: Hypomethylation of the perforin gene promoter in CD4 + T cells, inflammation and oxidative stress, might be involved in alveolar septal cell apoptosis associated with emphysema in rats. This study aimed to investigate the effects of S-adenosylmethionine (SAM) on this kind of apoptosis in rats with autoimmune emphysema. METHODS: Twenty-four rats were randomly divided into three groups: a normal control group, a model group, and a SAM group. Pathological changes in lung tissues were observed, and the mean linear intercept (MLI) and mean alveolar number (MAN) were measured. The levels of anti-endothelial cell antibodies (AECA) in serum, alveolar septal cell apoptosis, perforin gene promotor methylation in CD4 + T cells in the spleen, and the levels of cytokines, malondialdehyde (MDA), and glutathione (GSH) and the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in bronchoalveolar lavage fluid (BALF) were investigated. RESULTS: The MLI, apoptosis index (AI) of alveolar septal cells, levels of AECA in serum, and levels of tumour necrosis factor-α (TNF-α), matrix metalloproteinase-9 (MMP-9) and MDA in BALF were increased, while the MAN, methylation levels, and the activities of GSH, SOD and GSH-Px in BALF were decreased in the model group compared with those in the normal control group and the SAM group (all P < 0.05). The levels of interleukin-8 (IL-8) in BALF were greater in the model group than in the normal control group (P < 0.05). CONCLUSIONS: SAM protects against alveolar septal cell apoptosis, airway inflammation and oxidative stress in rats with autoimmune emphysema possibly by partly reversing the hypomethylation of the perforin gene promoter in CD4 + T cells.

Establishment of a leptospirosis model in guinea pigs using an epicutaneous inoculations route.

BACKGROUND: Leptospires are presumed to enter their host via small abrasions or breaches of the skin. The intraperitoneal route, although commonly used in guinea pig and hamster models of leptospirosis, does not reflect conditions encountered during natural infection. The aim of this study is to develop a novel leptospirosis guinea pig model through epicutaneous route and to elucidate the pathogenesis of leptospirosis in experimental guinea pigs by comparing the data from other studies using different infection routes. METHODS: The guinea pigs were inoculated with 5 × 108 Leptospira interrogans strain Lai onto either shaved-only or abraded skin. The guinea pigs were sacrificed at 2, 8, 24, 48, 72, 96 and 144 h post-infection (p.i.) followed by harvest of the lungs, liver, kidneys, spleen, and the skin around the inoculated sites for further examinations. Hematoxylin and eosin (HE) staining and electron microscopy were used to detect the pathologic changes. Real time PCR and immunohistochemistry staining were performed to detect dynamic distribution of leptospires in blood and tissues, respectively. RESULTS: In the guinea pigs with abraded skin inoculations, leptospires were detected in blood as early as 2 h post infection (p.i.) and then disseminated to the liver, lungs and kidneys of almost all animals within 96 h p.i.. Leptospires were also detected engulfed in the swelling vascular endothelial cells and were frequently aggregated around the capillaries in the dermis and subcutaneous tissue under the inoculated site. For the guinea pigs with abraded skin inoculations, hemorrhage at the dermis around the inoculated site was found before the appearance of internal organs hemorrhage, severe lesions such as hemorrhages in the lungs, nephritis, jaundice, haematuria were also observed, and two of seven guinea pigs died at 144 h p.i. while no lesions and leptospires were detected in the shaved-only guinea pigs using the same dose of strain Lai. CONCLUSION: Intact keratinocyte layer is a very efficient barrier against leptospires, and intact skin can prevent the infiltration of leptosipres to the host. Leptospires can penetrate abraded skin and quickly establish a systemic infection by crossing tissue barriers. We have successfully established a novel leptospirosis guinea pig model through epicutaneous inoculations route, which replicates a natural course of infection and appears to be an alternative way to investigate the pathogenesis of leptospirosis, especially in terms of early stage of host-pathogen interactions. This novel model may also be advantageous for studies of the mechanisms involved in cutaneous barriers and epidermal interactions with this organism.

[Clinical observation of different doses of continuous renal replacement therapy for severe pneumonia with acute kidney injury].

OBJECTIVE: To explore the efficacy of different doses of continuous renal replacement therapy (CRRT) in the treatment of severe pneumonia with acute kidney injury. METHODS: Twenty-eight patients with severe pneumonia and acute kidney injury were recruited from our hospital between February 2009 and March 2012. They divided into 3 groups: group A receiving a large dose of continuous veno-venous hemodiafiltration (CVVHDF) (70 ml×kg(-1)×h(-1), n = 9), group B a moderate dose of CVVHDF (45 ml×kg(-1)×h(-1), n = 8) and group C a low dose of CVVHDF (25 ml×kg(-1)×h(-1), n = 11). Before and after treatment, the changes of patient conditions, renal function and blood gas analysis were recorded. Such biomarkers as C-reactive protein (CRP), tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), interleukin-8 (IL-8), white blood cell (WBC) and neutrophil granulocyte (N) were determined. RESULTS: Compared with group C, the levels of leucocyte ((11.0 ± 3.2)×10(9)/L, (11.1 ± 5.3)×10(9)/L vs (8.5 ± 2.7)×10(9)/L), CRP ((89 ± 10), (90 ± 14) vs (107 ± 13) mg/L), TNF-α ((99 ± 39), (103 ± 28) vs (123 ± 35) pg/L), IL-6 ((54 ± 22), (69 ± 20) vs (81 ± 24) pg/L) and IL-8 ((104 ± 50), (138 ± 63) vs (167 ± 71) pg/L) decreased significantly in groups A and B after treatment (all P < 0.05). There were no differences in the levels of CRP, IL-6, IL-8 or TNF-α levels between groups B and C (all P > 0.05). The recovery of kidney function was much more rapid in group A than in groups B and C. CONCLUSION: The large dose of CRRT may be more effective in the clearance of inflammatory mediators and improved survival of severe pneumonia with acute kidney injury than moderate and low doses.

[Pollution Characteristics of Water-soluble Ions in PM2.5 During the Lantern Festival of 2021 in Zibo City].

Based on the online ion data, we have analyzed the cause of a PM2.5 pollution episode, which happened during the Lantern Festival in Zibo in 2021. The pollution characteristics of water-soluble ion components were analyzed, the formation mechanism of secondary inorganic ions (SNA) was discussed, and the changes in the liquid water content (LWC) and pH value of particulate matter before and after pollution were comparatively analyzed. The results showed that the pollution period before the Lantern Festival (T1) and the pollution period at night (T2) ρ(WSIIs) during the Lantern Festival were 46.83 µg·m-3 and 71.18 µg·m-3, respectively, which were 2.3 times and 3.6 times that of the cleaning period, respectively. Among them, the growth multiple of SNA during the T1 period was greater[ρ(NO3-) was 2.9 times, ρ(SO42-) was 2.8 times, and ρ(NH4+) was 2.4 times] than the growth multiple of PM2.5 (2.1 times), which showed that the increase in SNA concentration during the T1 period was the main reason for the increase in PM2.5 concentration. Furthermore, the Cl-, K+, and Mg2+ concentrations, which were 4.0, 14.8, and 16.5 times that of the cleaning period, respectively, increased significantly during the T2 period, indicating that the fireworks and firecrackers caused the rapid increase in the PM2.5 concentration during the T2 period. The LWC during the pollution period was 49.37 µg·m-3, which was 2.9 times that of the cleaning period. LWC was mainly affected by RH and NH4+ during the T1 period and was also affected by Mg2+ during the T2 period. The average pH during the pollution period in Zibo was 4.79±1.54, which was 0.14 lower than that during the cleaning period. The pH during the T1 period was affected by the combined effects of SO42- and NH4+, which made it decrease 0.53 compared to that during the cleaning period. The pH value during the T2 period may be affected by the K+, Cl-, and Mg2+ emitted from the fireworks and firecrackers, causing the pH to increase 0.65 compared to that during the cleaning period. The formation mechanism showed that SO42- was mainly generated by heterogeneous hydrolysis during the pollution episode, whereas NO3- was mainly generated by homogeneous reactions. On the whole, during the pollution episode, the increase in PM2.5 concentration before the Lantern Festival was mainly caused by the increase in SNA concentration, and the increase the night of the Lantern Festival was mainly caused by setting off fireworks and firecrackers.

Evaluation of a novel fusion protein antigen for rapid serodiagnosis of tuberculosis.

The detection of Mycobacterium tuberculosis-specific antibodies in human sera has been a rapid and important diagnostic aid for tuberculosis (TB) control and prevention. However, any single antigen is not enough to be used to cover the antibody profiles of all TB patients. In this study, a novel fusion protein was constructed using gene splicing by overlap extension (SOEing), and then the antibody level against it in 171 TB patients and 86 controls was evaluated by enzyme-linked immunosorbent assay. Compared with the three individual antigen (16 kDa: sensitivity 19.9%, specificity 96.5%; MPT64: sensitivity 75.4%, specificity 34.9%; 38 kDa: sensitivity 33.3%, specificity 83.7%), the fusion protein antigen (sensitivity 42.1%, specificity 89.5%) gave the best diagnostic performance with the largest receiver operating characteristic curve area 0.656 (95% confidence interval [CI], 0.590-0.721; P<0.01). These results suggested that the novel fusion protein antigen successfully constructed by gene SOEing provided the improved diagnostic performance for TB, and other mycobacterial multiepitope fusion proteins may also be worthy of investigation for further enhancing the detection sensitivity.

Unexplained elevation of erythrocyte sedimentation rate in a patient recovering from COVID-19: A case report.

BACKGROUND: A disease caused by a novel coronavirus virus, named coronavirus disease 2019 (COVID-19), broke out in Wuhan, China in December 2019, and spread around the word. As of March 4, 2020, 93090 confirmed cases and 2984 deaths have been reported in more than 80 countries and territories. It has triggered global public health security. However, the features and prognosis of COVID-19 are incompletely understood. CASE SUMMARY: We here report that the erythrocyte sedimentation rate (ESR) increased in a confirmed COVID patient. The high level of ESR sustained for a long time even after the patient recovered from COVID-19, while all results related to tumor, tuberculosis, rheumatic diseases, anemia, etc. cannot explain the abnormal elevation of ESR presented in this case. CONCLUSION: Although the increased ESR cannot be explained by all existing evidence, it possibly links the abnormal pathologic change in some COVID-19 patients and negative prognosis, and provides the clue to dissect the mechanism of illness progressing in COVID-19 and its prognosis.

A Low-Producing Haplotype of Interleukin-6 Disrupting CTCF Binding Is Protective against Severe COVID-19.

Interleukin6 (IL-6) is a key driver of hyperinflammation in COVID-19, and its level strongly correlates with disease progression. To investigate whether variability in COVID-19 severity partially results from differential IL-6 expression, functional single-nucleotide polymorphisms (SNPs) of IL-6 were determined in Chinese COVID-19 patients with mild or severe illness. An Asian-common IL-6 haplotype defined by promoter SNP rs1800796 and intronic SNPs rs1524107 and rs2066992 correlated with COVID-19 severity. Homozygote carriers of C-T-T variant haplotype were at lower risk of developing severe symptoms (odds ratio, 0.256; 95% confidence interval, 0.088 to 0.739; P = 0.007). This protective haplotype was associated with lower levels of IL-6 and its antisense long noncoding RNA IL-6-AS1 by cis-expression quantitative trait loci analysis. The differences in expression resulted from the disturbance of stimulus-dependent bidirectional transcription of the IL-6/IL-6-AS1 locus by the polymorphisms. The protective rs2066992-T allele disrupted a conserved CTCF-binding locus at the enhancer elements of IL-6-AS1, which transcribed antisense to IL-6 and induces IL-6 expression in inflammatory responses. As a result, carriers of the protective allele had significantly reduced IL-6-AS1 expression and attenuated IL-6 induction in response to acute inflammatory stimuli and viral infection. Intriguingly, this low-producing variant that is endemic to present-day Asia was found in early humans who had inhabited mainland Asia since ∼40,000 years ago but not in other ancient humans, such as Neanderthals and Denisovans. The present study suggests that an individual's IL-6 genotype underlies COVID-19 outcome and may be used to guide IL-6 blockade therapy in Asian patients. IMPORTANCE Overproduction of cytokine interleukin-6 (IL-6) is a hallmark of severe COVID-19 and is believed to play a critical role in exacerbating the excessive inflammatory response. Polymorphisms in IL-6 account for the variability of IL-6 expression and disparities in infectious diseases, but its contribution to the clinical presentation of COVID-19 has not been reported. Here, we investigated IL-6 polymorphisms in severe and mild cases of COVID-19 in a Chinese population. The variant haplotype C-T-T, represented by rs1800796, rs1524107, and rs2066992 at the IL-6 locus, was reduced in patients with severe illness; in contrast, carriers of the wild-type haplotype G-C-G had higher risk of severe illness. Mechanistically, the protective variant haplotype lost CTCF binding at the IL-6 intron and responded poorly to inflammatory stimuli, which may protect the carriers from hyperinflammation in response to acute SARS-CoV-2 infection. These results point out the possibility that IL-6 genotypes underlie the differential viral virulence during the outbreak of COVID-19. The risk loci we identified may serve as a genetic marker to screen high-risk COVID-19 patients.

Toxoplasma gondii infection in patients with lung diseases in Shandong province, eastern China.

Toxoplasma gondii, an intracellular protozoan parasite, can induce various clinical symptoms. T. gondii has been considered to play an important role in the pathogenesis of lung diseases. This survey was conducted to explore the correlation between T. gondii infection and lung diseases through a case-control study carried out in Shandong province, eastern China. In the present survey, T. gondii IgG antibodies were found in 76/398 (19.10%) of patients with lung diseases, which was significantly higher (P < 0.001) than the level found in the control subjects (35/398; 8.79%) through serological diagnosis. Patients with lung cancer have the highest T. gondii seroprevalence (26.19%), followed by Pulmonary cyst (25.00%), Tuberculosis (17.07%), Pneumonia (16.33%) and chronic obstructive pulmonary disease (COPD) (16.05%). Moreover, a semi-nest PCR targeted T. gondii B1 gene was employed to detect the T. gondii DNA in the blood samples. T. gondii DNA was detected in 5.53% blood samples of patients with lung diseases and 2.51% control subjects, respectively. The present study firstly shows that T. gondii has a high probability to infect the patients with lung diseases. Thus, the potential presence of T. gondii in patients with lung diseases should be appreciated during in the course of treatment and safeguard procedures should be implemented to protect vulnerable patients with lung diseases.

Indolent T cell lymphoproliferative disorder of the gastrointestinal tract: an uncommon case with lymph node involvement and the classic Hodgkin's lymphoma.

Here we describe an unusual case of an indolent CD8+ T-cell lymphoproliferative disorder in the gastrointestinal tract (ITCLD-GT) accompanied by neck lymph node infiltration and new onset of classic Hodgkin's lymphoma after two years follow-up. Previously, this 42-year-old Asian man suffered from diarrhea and abdominal pain for two years. Intestinal biopsies showed a group of small to intermediate-sized lymphocytes which were monomorphic as well as arranged in a nodular pattern with no clear boundary and were diagnosed as ITCLD-GT. He did not receive chemotherapy or have any disease progression in the gastrointestinal tract (GIT) during the follow-up until a development of neck lymphadenopathy, which led to an eventual mixed cellularity type of Hodgkin's lymphoma, one type of classic HL diagnosis. Interestingly, besides the Hodgkin's cells, the same pathological T-cells in the GIT were present in the Hodgkin's lymphoma lesions. These two pathological T cells in GIT and neck lymph node had the identical histopathological and molecular abnormalities that confirmed the abenteric distant infiltration of ITCLD-GT to the neck lymph node in this patient. This is the first case of ITCLD-GT that has a definite distant lymph node invasion. ITCLD-GT usually has a relatively good prognosis but patients with ITCLD-GT may have abenteric distant infiltration. Thus, long-term follow-up and further study of the underlying mechanisms of this process are necessary.

Impact of pulmonary interstitial lesions on efficacy and prognosis of EGFR-TKI-treated advanced non-small cell lung cancers.

BACKGROUND: This study aimed to assess the impact of pre-existing pulmonary interstitial lesions (PIL) on the efficacy and prognosis of patients with epidermal growth factor receptor (EGFR) mutant non-small cell lung cancer (NSCLC) treated with EGFR tyrosine kinase inhibitor (TKI). METHODS: Patients with advanced NSCLC harboring EGFR exon 19 deletion (E19 del) or exon 21 (E21) L858R were enrolled in this study. All patients underwent high resolution computed tomography (HRCT) chest scans prior to EGFR-TKI treatment. Pre-existing PIL was graded according to HRCT imaging (PIL 0, 1, 2, and 3). Cox proportional-hazards regression models were used to identify the prognostic factors for progression-free survival (PFS). RESULTS: A total of 134 eligible patients were enrolled. The overall objective response rate (ORR) and median PFS were 73.1% and 10.0 months (95% CI: 7.51-12.49), respectively. There were 62 (46.3%), 25 (18.7%), 28 (20.9%), and 19 (14.1%) cases of PIL grade 0, 1, 2, and 3, respectively, with median PFS and ORR of 12.9 months and 80.6%, 11.0 months and 72.0%, 10.0 months and 71.4%, and 7.0 months and 52.6%, respectively. Multivariate analysis showed that squamous cell carcinoma (vs. adenocarcinoma, HR =4.33), E21 L858R (vs. E19 del, HR =1.57), and PIL grade 3 (vs. grade 0-2, HR =1.60-2.48) were poor prognostic factors for PFS (P<0.05 for all). CONCLUSIONS: Pre-existing PIL grade is an independent prognostic factor for predicting resistance to EGFR-TKIs in patients with EGFR-mutant advanced NSCLC. Higher PIL grade suggests higher risk of early progression.

Molecular characterization of the pL40 protein in Leptospira interrogans.

Leptospirosis is a widespread zoonotic disease caused by pathogenic leptospires. The identification of outer membrane proteins (OMPs) conserved among pathogenic leptospires, which are exposed on the leptospiral surface and expressed during mammalian infection, has become a major focus of leptospirosis research. pL40, a 40 kDa protein coded by the LA3744 gene in Leptospira interrogans, was found to be unique to Leptospira. Triton X-114 fractionation and flow cytometry analyses indicate that pL40 is a component of the leptospiral outer membrane. The conservation of pL40 among Leptospira strains prevalent in China was confirmed by both Western blotting and PCR screening. Furthermore, the pL40 antigen could be recognized by sera from guinea pigs and mice infected with low-passage L. interrogans. These findings indicate that pL40 may serve as a useful serodiagnostic antigen and vaccine candidate for L. interrogans.

[Clinical investigation of the clinical management and therapeutic strategies to the first human case infected by influenza A/H5N1 in Guizhou Province].

OBJECTIVE: To explore the optimized clinical management and therapeutic strategies for the survived human case infected by influenza A (A/H5N1). METHODS: All the data of the first human case infected by A/H5N1 in Guizhou province was collected and analyzed. RESULTS: The first case infected by A/H5N1 in Guizhou Province was confirmed by laboratory findings with reverse-transcription polymerase chain reaction (RT-PCR) and A/H5N1 isolation. Patient was healthy in the past and exposed in the environment of living poultry. The initial symptoms was high fever without influenza-like presentation, but with extremity hyperspasmia and conscious disturbance sometimes. A productive cough with a large mount of pink foaming sputum then appeared. The clinical situation was rapidly deteriorated with dyspnea, acute respiratory distress syndrome and atrial fibrillation. Multiple infiltration in bilateral lungs was progressively developed with moderate bilateral pleural effusion. Invasive ventilation was intervened since ARDS on day 8 after sickness. Oseltamivir was kicked off since day 9 after sickness. However, the clinical condition was still exacerbated. High titering antibody of A/H5N1 vaccinated plasma was administrated on day 10 after sickness. The clinical condition (including oxygen saturation, respiratory symptoms, etc.) was improved rapidly. The weaning of ventilation was carried out in two days. Atrial fibrillation was back to normal. The patient was clinical recovery and was discharged from hospital on day 23 after sickness. CONCLUSIONS: The prognosis was poor if A/H5N1 infected human cases developed as acute respiratory distress syndrome with heart injury. However, it could be ameliorated if the plasma of A/H5N1 vaccinated neutralizing antibody was administrated in time or within two weeks after sickness.