Metatranscriptome of human lung microbial communities in a cohort of mechanically ventilated COVID-19 Omicron patients.

The Omicron variant of the severe acute respiratory syndrome coronavirus 2 (SARS­CoV­2) infected a substantial proportion of Chinese population, and understanding the factors underlying the severity of the disease and fatality is valuable for future prevention and clinical treatment. We recruited 64 patients with invasive ventilation for COVID-19 and performed metatranscriptomic sequencing to profile host transcriptomic profiles, plus viral, bacterial, and fungal content, as well as virulence factors and examined their relationships to 28-day mortality were examined. In addition, the bronchoalveolar lavage fluid (BALF) samples from invasive ventilated hospital/community-acquired pneumonia patients (HAP/CAP) sampled in 2019 were included for comparison. Genomic analysis revealed that all Omicron strains belong to BA.5 and BF.7 sub-lineages, with no difference in 28-day mortality between them. Compared to HAP/CAP cohort, invasive ventilated COVID-19 patients have distinct host transcriptomic and microbial signatures in the lower respiratory tract; and in the COVID-19 non-survivors, we found significantly lower gene expressions in pathways related viral processes and positive regulation of protein localization to plasma membrane, higher abundance of opportunistic pathogens including bacterial Alloprevotella, Caulobacter, Escherichia-Shigella, Ralstonia and fungal Aspergillus sydowii and Penicillium rubens. Correlational analysis further revealed significant associations between host immune responses and microbial compositions, besides synergy within viral, bacterial, and fungal pathogens. Our study presents the relationships of lower respiratory tract microbiome and transcriptome in invasive ventilated COVID-19 patients, providing the basis for future clinical treatment and reduction of fatality.

Post-harvest ripening affects drying behavior, antioxidant capacity and flavor release of peach via alteration of cell wall polysaccharides content and nanostructures, water distribution and status.

Effect of post-harvest ripening on cell wall polysaccharides nanostructures, water status, physiochemical properties of peaches and drying behavior under hot air-infrared drying was evaluated. Results showed that the content of water soluble pectins (WSP) increased by 94 %, while the contents of chelate-soluble pectins (CSP), Na2CO3-soluble pectins (NSP) and hemicelluloses (HE) decreased during post-harvest ripening by 60 %, 43 %, and 61 %, respectively. The drying time increased from 3.5 to 5.5 h when the post-harvest time increased from 0 to 6 days. Atomic force microscope analysis showed that depolymerization of hemicelluloses and pectin occurred during post-harvest ripening. Time Domain -NMR observations indicated that reorganization of cell wall polysaccharides nanostructure changed water spatial distribution and cell internal structure, facilitated moisture migration, and affected antioxidant capacity of peaches during drying. This leads to the redistribution of flavor substances (heptanal, n-nonanal dimer and n-nonanal monomer). The current work elucidates the effect of post-harvest ripening on the physiochemical properties and drying behavior of peaches.

Effects of hot-air drying temperature on drying characteristics and color deterioration of rape bee pollen.

The effects of different hot-air drying (HAD) temperature (40, 50, 60, and 70 °C) on the drying characteristics, color changes, the contents of α-dicarbonyl compounds (α-DCs), 5-hydroxymethyl furfural (5-HMF) and carotenoids of rape bee pollen were investigated in the study. The results showed that increasing the drying temperature from 40 to 70 °C shortened the drying time by 65 %. HAD caused lower L* and b* values, as well as higher a* values. Browning index and 5-HMF content increased with increasing drying temperature. The relative content of antheraxanthin increased 230 % at 70 °C while lutein and zeaxanthin decreased by 74 and 81 % than that of fresh (non-heated) pollen. The contents of 3-deoxyglucosone, 1-deoxy-2,3-pentosulose, antheraxanthin, and lutein were related to the color deterioration in HAD process in rape bee pollen. This work is of great practical significance to provide scientific basis for quality optimization of bee pollen in the drying process.

EFEMP2 Suppresses the Invasion of Lung Cancer Cells by Inhibiting Epithelial-Mesenchymal Transition (EMT) and Down-Regulating MMPs.

BACKGROUND: Epidermal growth factor-containing fibulin-like extracellular matrix protein 2 (EFEMP2), also known as fibulin-4, MBP1 and UPH1, is an extracellular matrix protein associated with a variety of tumors. The purpose of this study was to investigate the prognostic value and the function of EFEMP2 in lung cancer. METHODS: The mRNA and protein expression of EFEMP2 in lung normal and cancer tissues, lung cancer cell lines (A549, H460, H1299 and H1650) and normal epithelial cell line BEAS-2B were evaluated by immunohistochemistry, RT-qPCR and Western blotting. The Public databases (Oncomine and Kaplan-Meier plotter) were used to investigate the prognostic value of EFEMP2 in lung cancer. RNA interference (RNAi) and overexpression transfection were performed to detect the effects of EFEMP2 up- or down-regulation on lung normal and cancer cell proliferation, invasion and metastasis in vitro and in vivo. RESULTS: EFEMP2 was lowly expressed in lung cancer tissues and cells, and its low expression was associated with malignant phenotype and poor prognosis of lung cancer. The same conclusion had been drawn from the Public databases. EFEMP2 overexpression significantly inhibited the invasion of lung cancer cells, hampered the process of EMT, and decreased the expression and activity of MMP2 and MMP9, while EFEMP2 knockdown remarkably enhanced the invasion of lung cancer cells, promoted EMT, and increased the expression and activity of MMP2 and MMP9. CONCLUSION: The low expression of EFEMP2 was detected in lung cancer and was positively correlated with the poor prognosis of patients. EFEMP2 was a tumor suppressor gene that inhibited the progress of lung cancer, which suggested a new research objective for the future studies.

The association between oxidized low-density lipoprotein antibodies and hematological diseases.

BACKGROUND: The aim of the study is to compare the profiles of antibodies (IgM and IgG) against oxidized low-density lipoprotein (oxLDL) of hematological diseases. METHODS: The serum antibodies of oxLDL-IgM and oxLDL-IgG for 446 cases with hematological diseases and 90 patients with primary hypertension and 90 healthy controls were measured by enzyme-linked immunosorbent assay (ELISA) in a cross-section survey. The association of serum oxLDL-LgM and oxLDL-IgG with hematological diseases was analyzed by multiple linear regression model. RESULTS: Comparing with the hypertension or normal groups, the levels of TCH, TG, LDL-c, HDL-c, oxLDL, and oxLDL-IgG were lower and the levels of ADP and oxLDL-IgM were higher in the hematological diseases group. The levels of oxLDL-IgG antibodies titer were different among hematological diseases group. The results of correlation and multiple regression analysis showed that the seven hematological disease subgroups were positively related to the oxLDL-IgM antibody titer but negatively related to the oxLDL-IgG antibody titer, having been adjusted for potential confounding factors such as age, SBP, DBP, BMI, TCH, TG, ADP, oxLDL, HDL-c, LDL-C. CONCLUSIONS: Here we show that oxLDL-IgG antibodies titer were lower and of oxLDL-IgM titer were higher than hypertension and healthy individuals. Also oxLDL-IgG titer were different among hematological diseases group.

Effects of periodontal therapy on serum lipid profile and proinflammatory cytokines in patients with hyperlipidemia: a randomized controlled trial.

OBJECTIVE: The aim of this study was to evaluate whether periodontal treatment in patients with periodontitis and hyperlipidemia may have any influence on plasma lipids and pro-inflammatory cytokine levels. MATERIAL AND METHODS: We randomly assigned 109 patients with hyperlipidemia and chronic periodontitis into group 1 (n = 55) and group 2 (n = 54). Patients in group 1 underwent a standard cycle of supragingival mechanical scaling and polishing. Patients in group 2 underwent the adjunctive full-mouth intensive removal of subgingival dental plaque biofilms with the use of scaling and root planning. Periodontal parameters, total cholesterol (TC), triglyceride (TRG), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), tumor necrosis factor-alpha (TNF-α), interleukin(IL)-1ß(IL-1ß), and IL-6 were evaluated before treatment and 2 and 6 months after treatment. RESULTS: Two and 6 months after treatment, TRG levels were significantly lower in group 2 than in group 1 (P < 0.05), and the levels of HDL-C were significantly higher (P < 0.05). Two and 6 months after therapy, the levels of TNF-α were significantly lower in group 2 than in group 1 (P < 0.05), as were the levels of IL-1ß (P < 0.001) and IL-6 (P < 0.001). CONCLUSIONS: Intensive periodontal treatment of participants with hyperlipidemia and chronic periodontitis improved serum lipid levels and decreased circulating pro-inflammatory cytokine levels. CLINICAL RELEVANCE: This study showed that intensive treatment of periodontitis results in an improvement in serum lipid levels and a decrease in serum proinflammatory cytokine levels in patients with periodontitis and hyperlipidemia. These findings may contribute to present knowledge that periodontal therapy may be beneficial for individuals with hyperlipidemia.

The anti-leukemic effect of carnosic acid combined with adriamycin in a K562/A02/SCID leukemia mouse model.

The effects of carnosic acid (CA) were investigated on the acute myeloid leukemia (AML) cell growth in vivo. A NOD/SCID AML mouse model, which was set up by inoculation with K562/A02 cells, was used to study whether tumor growth in vivo can be inhibited by CA combined with adriamycin. After being inoculated with K562/A02 cells, the NOD/SCID mice were expressed positive human mdr1 and bcr/abl genes. This result indicates that the K562/A02/SCID leukemia mouse model is successfully established. The mice treated with CA combined with adriamycin exhibit a significant lower number of leukemia cells (20%) than that of untreated animals (32.5%) (P<0.05), in particular with higher percentages of apoptotic cells than the mice treated by single adriamycin (control) group. The median of 95% CI survival time is 19 (10.0-44.2) and 33 (29.4-36.6) days for the control group and the CA-treated group, respectively. The difference is statistically significant (P<0.05). It is illustrated that the natural compound CA, combined with Adriamycin, has high potential to inhibit the growth of malignant cells in vivo, and is a promising adjuvant anti-cancer drug. Prospective studies should be conducted to understand the functional mechanism of CA at the molecular level.

[Valacyclovir as an adjunct to full-mouth scaling and root planing of advanced chronic periodontitis:a randomized clinical trail].

PURPOSE: The aim of this study was to evaluate the clinical benefit of valacyclovir when performing full-mouth periodontal debridement in patients with advanced chronic periodontitis. METHODS: Fifty-nine patients with advanced chronic periodontitis were randomly assigned into control-treatment group(n=29) and intensive-treatment group(n=30). All patients were given instructions of basic oral hygiene and a standard cycle of supragingival mechanical scaling and polishing. Patients in the intensive-treatment group received oral valacyclovir for 1 week, while patients in the control-treatment group received placebo. Thereafter, patients in both groups underwent full-mouth intensive removal of subgingival dental plaque biofilms with the use of scaling and root planing within 48 hours. Periodontal parameters were evaluated before treatment and 2 or 6 months after treatment. The data was statistically analyzed using SPSS17.0 software package. RESULTS: No significant difference in clinical parameters was noted before treatment. 2 and 6 months after treatment, the mean percentage reduction of sites with BOP and PD≥4 mm were significantly higher in the intensive-treatment group than in the control-treatment group (P<0.05). Similarly, patients in the intensive-treatment group had higher mean PD reduction than those in the control-treatment group 2 months (P<0.05) and 6 months after therapy (P<0.05). However, the mean values of CAL reduction were slightly and not significantly higher in the intensive-treatment group than in the control-treatment group after therapy. CONCLUSIONS: It may be concluded that valacyclovir significantly improves clinical results of full-mouth non-surgical periodontal debridement in advanced chronic periodontitis.

The relationship between KRAS gene mutations and HLA class I antigen downregulation in the metastasis of non-small cell lung cancer.

OBJECTIVE: To investigate the association between v-Ki-ras2 Kirsten rat sarcoma viral oncogene homologue (KRAS) gene mutations and levels of human leucocyte antigen (HLA) class I antigen in primary lung tumours and metastatic lymph nodes of patients with non-small cell lung cancer (NSCLC). METHODS: Patients with NSCLC undergoing tumour resection were enrolled. KRAS codon 12 mutations were analysed in normal lung and lymph node tissue, primary lung tumours and metastatic lymph nodes using polymerase chain reaction-restriction fragment length polymorphism analysis. HLA class I antigen immunostaining was examined using flow cytometry. RESULTS: A total of 65 patients participated in the study. All normal lung tissues had positive HLA class I antigen immunostaining. The majority of primary lung tumours (56/65) and all of the metastatic lymph nodes (31/31) had downregulated HLA class I antigen immunostaining. There was a positive correlation between downregulated HLA class I antigen in primary tumours and metastatic lymph nodes. There was a negative correlation between KRAS codon 12 mutations and the level of HLA class I antigen in primary and metastatic tumours. CONCLUSIONS: KRAS codon 12 mutations appear to be important in the downregulation of HLA class I antigen in NSCLC. Abnormal activation of the oncogenic KRAS pathway might provide a new treatment target for NSCLC.

Synergism between carnosic acid and arsenic trioxide on induction of acute myeloid leukemia cell apoptosis is associated with modulation of PTEN/Akt signaling pathway.

OBJECTIVE: To investigate the synergistic effects of carnosic acid (CA) with arsenic trioxide (As2O3) on proliferation and apoptosis in HL-60 human myeloid leukemia cells, and the major cellular signaling pathway involved in these effects. METHODS: HL-60 cellular proliferation was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) analysis. Cell cycle distribution and apoptosis were monitored by flow cytometry. The activation of casepase-9, Bcl-2-associated agonist of cell death (BAD), p-BAD, p27, phosphatase and tensin homolog deleted on chromosome ten (PTEN), Akt, p-Akt was assessed by Western blot analysis. The expression of PTEN mRNA was tested by reverse transcription polymerase chain reaction (RT-PCR) analysis. RESULTS: CA reduced HL-60 cell viability in a dose- and time-dependent manner, and induced G1 arrest and apoptosis. Moreover, CA upregulated PTEN expression, blocked the Akt signaling pathway, subsequently inhibited phosphorylation of BAD, reactivated caspase-9, and elevated levels of p27. CA also augmented these effects of As2O3. CONCLUSION: CA might be a novel candidate of the combination therapy for leukemia treatment; these effects were apparently associated with the modulation of PTEN/Akt signaling pathway.

[Relationship of TGF-ß and IL-4R gene polymorphisms with risk of classical Hodgkin lymphoma].

OBJECTIVE: This study was aimed to analyze the relationship between single nucleotide polymorphisms of transforming growth factor-ß1 G-800A and C-509T, interleukin-4 receptor V75I and susceptibility of CHL in adults. METHODS: Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was applied to analyze the expressed alleles of the selected SNP loca. The relationship between genomic polymorphisms of TGF-ß1 and IL-4R and susceptibility of CHL were coupled with clinical data. RESULTS: TGF-ß1G-800A and TGF-ß1C-509T had obvious linkage equilibrium (D' = 0.879, r(2) = 0.83, P = 0.020). GT haplotype distribution frequencies in mixed cellularity Hodgkin lymphoma cases and control group were of 53.1% and 34.2%, respectively, with statistically significant (OR = 2.35, P = 0.000); distribution frequencies of mutant gene T/T in disease and control groups were of 38.8% and 15.3%, respectively, also with statistically significant (OR = 3.654, P = 0.000); frequencies of nodular sclerosis CHL patients with IL-4R V75I mutant gene A/A in disease and control groups were of 19.2% and 41.75%, respectively, also with statistically significant (OR = 3.156, P = 0.000). CONCLUSION: Single nucleotide polymorphisms of TGF-ß1 G-800A, C-509T and IL-4R V75I has a significant correlation with Chinese susceptibility to classical Hodgkin lymphoma.

[Study on reversing mechanism of multidrug resistance of K562/A02 cell line by carnosic acid].

OBJECTIVE: To investigate the effects of carnosic acid (CA) on reversal of the multidrug resistance (MDR) of human leukemia cell line K562/A02 and its mechanism. METHODS: MTT assay was used to determine the sensitivity of K562/A02 cells to adriamycin (ADM) pre-and post-treated with CA. Flow cytometry (FCM) and laser scanning confocal microscopy (LSCM) were used to measure intracellular fluorescence intensity and concentration of ADM respectively. The expression level of mdr1 was detected by semi-quantitative RT-PCR. P-glycoprotein (P-gp) expression was detected by FCM and Western blot. RESULTS: CA decreased IC(50) of ADM in K562/A02 cells from 16.31 µg/mL to 1.35 µg/mL, being a 12.08-fold decrease. The intracellular ADM fluorescence intensity of K562/A02 was increased from 17.05 to 60.53 after treated with CA (P < 0.01). In living K562/A02 cells, after treated with CA, the diffuse distribution of intracellular ADM was recovered in both nuclear and cytoplasm, and the concentration of intracellular ADM increased from 4.93µg/mL to 15.43µg/mL. RT-PCR assay showed that CA inhibited the expressions of mdr1 mRNA in K562/A02 cells (P < 0.01). Mean fluorescence intensity of P-gp detected by FCM in CA-treated K562/A02 was decreased to 22.80 as compared with that in untreated K562/A02 cells (44.40, P < 0.05). CONCLUSION: CA can reverse the MDR of K562/A02 cells in vitro. The mechanism may be associated with down-regulation of mdr1 and inhibition of P-gp function.

Reversion of P-glycoprotein-mediated multidrug resistance in human leukemic cell line by carnosic acid.

One of the common hindrances to successful chemotherapy is the development of multidrug resistance (MDR) by tumor cells to multiple chemotherapeutic agents. In this regard, P-glycoprotein (P-gp) acts as an energized drug pump that reduces the intracellular concentration of drugs, even of structurally unrelated ones. The modulators of P-gp function can restore the sensitivity of MDR cells to anticancer drugs. Therefore, to develop effective drug-resistance-reversing agents, we evaluated the P-gp modulating potential of carnosic acid (CA) in multidrug-resistant K562/AO2 cells in the present study. The reversing effect of CA was evaluated by determining the inhibition rates of cell viability with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide (MTT) assays. The intracellular adriamycin fluorescence intensity and the expression of P-gp were measured by flow cytometry (FCM). Meanwhile, the subcellular distribution of adriamycin was detected via Laser Scanning Confocal Microscopy (LSCM). The mRNA expression of mdrlwas then detected via semiquantitative reverse transcription polymerase chain reaction (RT-PCR). The findings showed that CA decreased apparently the Inhibition Concentration 50% (IC50) of adriamycin by increasing its intracellular concentration and thus enhancing the sensitivity of K562/AO2 cells. Adriamycin was distributed evenly in the cytoplasm when the cells were treated with CA. The expression of mdrl was decreased. Overall, the results indicated that CA can serve as a novel, non-toxic modulator of MDR, and it can reverse the MDR of K562/AO2 cells in vitro by increasing intracellular adriamycin concentration, down-regulating the expression of mdrl, and inhibiting the function of P-gp.

[Precipitation trapping with phenylfluorone and determination of trace gallium, germanium, molybdenum and indium by GFAAS].

The preconcentration of trace gallium, germanium, molybdenum and indium by trapping with precipitation of phenylfluorone (PF), and the determination of the elements by GFAAS were developed. The effects such as those of acidity, amounts of PF, aging time, volume of test solution, and the coexistent ions on the preconcentration of the trace elements were examined in detail. The optimum conditions of preconcentration for Ga(III) were pH approximately 2 test solution 500 mL with added 10.00 mg x mL(-1) PF (2.00 mL) and aging for 4 h, those for Ge(IV) were pH approximately 2 test solution 500 mL with added 10.00 mg x mL(-1) PF (4.00 mL) and aging for 10 h, those for Mo(V) were pH approximately 3 test solution 1 000 mL with added 10.00 mg x mL(-1) PF (3.00 mL) and aging for 6 h, and those for In(III) were pH approximately 5 test solution 100 mL with added 10.00 mg x mL(-1) PF (3.00 mL) and aging for 10 h. The experiment results showed that the main contribution to trapping trace gallium, germanium, molybdenum and indium with PF precipitation was post-precipitation instead of coprecipitation. The detection limits (3s) were 0.12 ng x mL(-1) for gallium, 0.30 ng x mL(-1) for germanium, 0.046 ng x mL(-1) for molybdenum and 2.7 ng x mL(-1) for indium. The developed methods were successfully applied to the determination of trace amount of the elements in water samples, geological standard reference materials, and zinc concentrate samples by graphite furnace atomic absorption spectrometry.

[Experimental study on immunotherapy with dendritic cell in leukemic mice model].

OBJECTIVE: To explore the feasibility and efficiency of immunotherapy with dendritic cell (DC) in leukemic mice model after allogeneic bone marrow transplantation (allo-BMT). METHODS: Mature DC were expanded from mice bone marrow mononuclear cells (MNC) by adding mouse granulocyte-macrophage colony stimulating factor (mGM-CSF) and interleukin-4 (mIL-4). Three days later they were pulsed with frozen thawing L7212 leukemia-related antigen. Mice bearing leukemia received allo-BMT at d 0, and then were divided into control group (A), T cells group (B) and DC + T cells group (C) to receive respective immune therapy at d 14. The survival rate, survival time, occurrence of graft-versus-host disease (GVHD), cytotoxicity of spleen cells and serum cytokine level were observed. The survivors in each group were rechallenged with L7212 cells to observe the immune response to the leukemia. RESULTS: Mature DC were successfully induced from bone marrow MNC. In groups B and C, the relapse rates were 30% and 0%, while the long term survival rates after BMT was 30% and 70% respectively. Both of the differences were statistically significant (P < 0.05). However, the incidence of GVHD in these two groups were similar. The mean survival times were (32.95 +/- 13.29) days and (41.15 +/- 13.88) days, respectively (P < 0.01). MTT assay indicated that spleen cells from group C had specific killing activity to L7212 cells. Enzyme-labeled immunosorbent assay (ELISA) showed that the serum IL-2 level in group C was (419.75 +/- 26.66) pg/ml, being significantly higher than that in the other two groups (P < 0.01). When the survivors were rechallenged with L7212 cells, there was difference between the survival rates of groups C and B (85.7% vs 33.3%, P < 0.05). CONCLUSION: Immunotherapy with leukemia related antigen-pulsed DC in combination with donor lymphocyte infusions is an effective approach to reinforce GVL effect and decrease relapse after allo-BMT.

[A randomized controlled and multicenter clinical study of ferrous L-threonate in treatment of iron deficiency anemia].

OBJECTIVE: To study the clinical efficacy and side effects of ferrous L-threonate for treatment of iron deficiency anemia (IDA). METHODS: It is a multicentral, randomized, double blind, double placebo and paralled comparative study with positive control. One hundred and forty IDA patients diagnosed according to the standard criteria in three hospitals were randomly divided into a test group (ferrous L-threonate plus placebo ferrous succinate) and a positive control group (ferrous succinate plus placebo ferrous L-threonate). Some iron parameters were examined 1, 4 and 8 weeks after medication. Hemoglobin, reticulocyte and other parameters for safety observation were collected every two weeks. RESULTS: For the 2 groups, self comparison showed significant difference (P < 0.01). The total efficacy is 98.44% and 97.01% respectively with no difference. Hemoglobin rised rapidly and gradually and reached a peak in week 8, the change was statistically significant (P < 0.01). Changes of iron parameters also showed significant difference. Side-effects were similar in both groups (13.85% and 14.71%, P > 0.05). CONCLUSION: The effect of ferrous L-threonate in IDA treatment is significant and rapid. Side-effects are few and minimal.