Proteomic investigation and biomarker identification of lung and spleen deficiency syndrome in HIV/AIDS immunological nonresponders.

Background: Human immunodeficiency virus (HIV) and acquired immune deficiency syndrome (AIDS) immunological nonresponders (HIV/AIDS-INRs) whose CD4+ cell counts do not rebound after highly active antiretroviral therapy (HAART) treatment usually experience severely impaired immune function and high mortality. Traditional Chinese medicine (TCM) has many advantages in the field of AIDS, especially its promotion of patients' immune reconstitution. Accurate differentiation of TCM syndromes is a prerequisite for guiding an effective TCM prescription. However, the objective and biological evidence for identification of the TCM syndromes in HIV/AIDS-INRs remains lacking. Lung and spleen deficiency (LSD) syndrome, a typical HIV/AIDS-INR syndrome, was examined on in this study. Methods: We first performed a proteomic study of LSD syndrome in INRs (INRs-LSD) using tandem mass tag combined with liquid chromatography-tandem mass spectrometry (TMT-LC-MS/MS) and screened them against the healthy and undocumented identifiable groups. The TCM syndrome-specific proteins were subsequently validated based on bioinformatics analysis and enzyme-linked immunosorbent assay (ELISA). Results: A total of 22 differentially expressed proteins (DEPs) were screened in INRs-LSD compared to the healthy group. Based on bioinformatic analysis, these DEPs were found to be mainly associated with the immunoglobin A (IgA)-generated intestinal immune network. In addition, we examined the TCM syndrome-specific proteins alpha-2-macroglobulin (A2M) and human selectin L (SELL) with ELISA and found that they were both upregulated, which was consistent with the proteomic screening results. Conclusions: A2M and SELL were finally identified as potential biomarkers for INRs-LSD, providing a scientific and biological basis for identifying typical TCM syndromes in HIV/AIDS-INRs and an opportunity to build a more effective TCM treatment system for HIV/AIDS-INRs.

The influence of physical exercise on college students' life satisfaction: The chain mediating role of self-control and psychological distress.

Objective: To explore the relationship between physical exercise and life satisfaction among college students and test the dual mediating role of self-control and psychological distress between them. Methods: A sample of 526 Chinese college students completed questionnaires regarding physical exercise, life satisfaction, self-control and psychological distress, of which 38.4% were boys. Results: Path analyzes indicated that physical exercise was positively correlated with life satisfaction, and this link could be mediated by self-control and psychological distress. Conclusion: The present study identifies the potential underlying mechanism by which physical exercise is associated with the life satisfaction of college students, which has important implications for theory and prevention.

Epidemiological investigation and proteomic profiling of typical TCM syndrome in HIV/AIDS immunological nonresponders.

HIV/AIDS pandemic remains the world's most severe public health challenge, especially for HIV/AIDS immunological nonresponders (HIV/AIDS-INRs), who tend to have higher mortality. Due to the advantages in promoting patients' immune reconstitution, Traditional Chinese medicine (TCM) has become one of the mainstays of complementary treatments for HIV/AIDS-INRs. Given that effective TCM treatments largely depend on precise syndrome differentiation, there is an increasing interest in exploring biological evidence for the classification of TCM syndromes in HIV/AIDS-INRs. In our study, to identify the typical HIV/AIDS-INRs syndrome, an epidemiological survey was first conducted in the Liangshan prefecture (China), a high HIV/AIDS prevalence region. The key TCM syndrome, Yang deficiency of spleen and kidney (YDSK), was evaluated by using a tandem mass tag combined with liquid chromatography-tandem mass spectrometry (TMT-LC-MS/MS). A total of 62 differentially expressed proteins (DEPs) of YDSK syndrome compared with healthy people were screened out. Comparative bioinformatics analyses showed that DEPs in YDSK syndrome were mainly associated with response to wounding and acute inflammatory response in the biological process. The pathway annotation is mainly enriched in complement and coagulation cascades. Finally, the YDSK syndrome-specific DEPs such as HP and S100A9 were verified by ELISA, and confirmed as potential biomarkers for YDSK syndrome. Our study may lay the biological and scientific basis for the specificity of TCM syndromes in HIV/AIDs-INRs, and may provide more opportunities for the deep understanding of TCM syndromes and the developing more effective and stable TCM treatment for HIV/AIDS-INRs.

Anti-hepatocellular carcinoma efficacy of Fuzheng Xiaozheng prescription and its interventional mechanism studies.

ETHNOPHARMACOLOGICAL RELEVANCE: Fuzheng Xiaozheng prescription (FZXZP), a traditional Chinese medicine, which was derived from the famous decoction, Sanjiasan, in the book of "Wenyilun" in Ming dynasty. Due to its function of invigorating the circulation of blood in Chinese medicine, it was usually used for treating the liver cirrhosis, hepatocellular carcinoma (HCC), etc. Clinical application found that FZXZP exhibited satisfactory therapeutic effects in HCC treatments. However, we still know little about the underlying mechanisms. AIM OF STUDY: In this study, we aim to gain a deeper insight into the inhibiting effects of FZXZP on HCC rats and preliminarily elucidate the underlying intervention effects. MATERIALS AND METHODS: Two doses of FZXZP were adopted to evaluate the therapeutic effects on rat HCC, and then the intervention effects were evaluated from different aspects. High performance liquid chromatography (HPLC) was used for the active compounds prediction in FZXZP. Finally, the mRNA-Seq was conducted to reveal the intervention mechanisms and the mechanisms were further validated by quantitative Real-time PCR (qRT-PCR) and lipid contents analyses. RESULTS: The results showed that FZXZP significantly alleviated the serum biochemical indicators and improved the pathological characteristics of HCC rats. Mechanistically, FZXZP could regulate some lipid related metabolisms, including arachidonic acid, linoleic acid and retinol, as well as improving the steroid hormone biosynthesis, to improve the inflammatory statuses and restoring ability of HCC livers, and these were further confirmed by our following analyses on serum lipid contents and cytokine expressions. In addition, FZXZP could also negatively regulate four extracellular growth factors which could result in the blocking of two cancer-related signaling pathways, Ras/MAPK and Ras/PI3K-Akt. CONCLUSION: Our results suggested that FZXZP demonstrated significant inhibiting effects on rat HCC progresses, and these may be realized by improving the inflammatory statuses and blocking the Ras/MAPK and Ras/PI3K-Akt signaling pathways.

Comparative serum microRNA array analysis of the spleen-stomach dampness-heat syndrome in different diseases: Chronic hepatitis B and chronic gastritis.

Spleen-stomach dampness-heat syndrome (SSDHS) is the common Traditional Chinese Medicine (TCM) syndrome observed in both chronic hepatitis B (CHB) and chronic gastritis (CG). The specialized TCM prescription for CHB and CG patients with SSDHS is same, but there is limited information about the biological characteristics of this TCM syndrome. This study aimed to identify the serum miRNAs profile for the SSDHS in two different diseases in order to evaluate the miRNA-mediated biological characteristics of this TCM syndrome. We performed comparative microarray analysis of serum miRNA expression profiles in 10 CHB patients with SSDHS (SSDHS-CHB), 10 CG patients with SSDHS (SSDHS-CG), and 10 healthy controls (HC). The selected miRNAs were further validated by qRT-PCR in 13 SSDHS-CHB patients, 13 SSDHS-CG patients, and 13 HC. Moreover, bioinformatics analysis (GO and KEGG pathway enrichment analyses) was applied to identify the involved target genes and pathways for these selected miRNAs. Nine significantly differentially expressed (SDE)-miRNAs in the SSDHS-CHB group and 24 SDE-miRNAs in the SSDHS-CG group were identified, compared with the HC group (fold change >2.0 and p < .05). Among these, upregulated hsa-miR-483-3p and downregulated hsa-miR-223-3p were identified as the common SDE-miRNAs for both SSDHS-CHB and SSDHS-CG groups. Bioinformatics analysis of the common SDE-miRNA's target genes showed their involvement in the regulation of inflammation, immune response, and tumorigenesis. SSDHS-specific hsa-miR-483-3p and hsa-miR-223-3p identified in this study indicated a relevance to the underlying biological basis of SSDHS, and may provide scientific basis for the application of same TCM prescription in CHB and CG.

Design, synthesis, and biological evaluation of nitroisoxazole-containing spiro[pyrrolidin-oxindole] derivatives as novel glutathione peroxidase 4/mouse double minute 2 dual inhibitors that inhibit breast adenocarcinoma cell proliferation.

A series of highly active CF3-containing 3'-(nitroisoxazole)spiro[pyrrolidin-3,2'-oxindoles] were synthesized and found to be novel glutathione peroxidase 4 (GPX4)/mouse double minute 2 (MDM2) dual inhibitors. Bioactive spirooxindole and isoxazole skeletons were combined, and the resulting compounds exhibited strong activities against both targets. In particular, compound 3d displayed excellent activity in the suppression of MDM2-mediated degradation of p53, as well as levels of GPX4, in MCF-7 breast cancer cells. Moreover, 3d also exhibited inhibitory effects on MDM2 and GPX4 in MCF-7 xenograft model to trigger ferroptotic and apoptotic cell death in in vivo experiments, which was consistent with the results of in vitro experiments.

Biological Differentiation of Dampness-Heat Syndromes in Chronic Hepatitis B: From Comparative MicroRNA Microarray Profiling to Biomarker Identification.

Increasing interest is aroused by traditional Chinese medicine (TCM) treatment of chronic hepatitis B (CHB) based on specific TCM syndrome. As the most common CHB syndromes, spleen-stomach dampness-heat (SSDH) syndrome and liver-gallbladder dampness-heat (LGDH) syndrome are still apt to be confused in TCM diagnosis, greatly hindering the stable exertion of TCM effectiveness. It is urgently needed to provide objective and biological evidences for differentiation and identification of the two significant syndromes. In this study, microRNA (miRNA) microarray analyses coupled with bioinformatics were employed for comparative miRNA profiling of SSDH and LGDH patients. It was found that the two syndromes had both the same and different significantly differentially expressed miRNAs (SDE-miRNAs). Commonness and specificity were also both found between their SDE-miRNA-based bioinformatics analyses, including Hierarchical Clustering, Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways, and miRNA-GO/pathway networks. Furthermore, syndrome-specific SDE-miRNAs were identified as the potential biomarkers, including hsa-miR-1273g-3p and hsa-miR-4419b for SSDH as well as hsa-miR-129-1-3p and hsa-miR-129-2-3p for LGDH. All these laid biological and clinical bases for classification and diagnosis of the two significant CHB dampness-heat syndromes including SSDH and LGDH, providing more opportunities for better application of TCM efficacy and superiority in CHB treatment.

Comparative Proteomic Profiling and Biomarker Identification of Traditional Chinese Medicine-Based HIV/AIDS Syndromes.

Given the challenges in exploring lifelong therapy with little side effect for human immunodeficiency virus infection and acquired immune deficiency syndrome (HIV/AIDS) cases, there is increasing interest in developing traditional Chinese medicine (TCM) treatments based on specific TCM syndrome. However, there are few objective and biological evidences for classification and diagnosis of HIV/AIDS TCM syndromes to date. In this study, iTRAQ-2DLC-MS/MS coupled with bioinformatics were firstly employed for comparative proteomic profiling of top popular TCM syndromes of HIV/AIDS: accumulation of heat-toxicity (AHT) and Yang deficiency of spleen and kidney (YDSK). It was found that for the two TCM syndromes, the identified differential expressed proteins (DEPs) as well as their biological function distributions and participation in signaling pathways were significantly different, providing biological evidence for the classification of HIV/AIDS TCM syndromes. Furthermore, the TCM syndrome-specific DEPs were confirmed as biomarkers based on western blot analyses, including FN1, GPX3, KRT10 for AHT and RBP4, ApoE, KNG1 for YDSK. These biomarkers also biologically linked with the specific TCM syndrome closely. Thus the clinical and biological basis for differentiation and diagnosis of HIV/AIDs TCM syndromes were provided for the first time, providing more opportunities for stable exertion and better application of TCM efficacy and superiority in HIV/AIDS treatment.

Comparison of the pulmonary dead-space fraction derived from ventilator volumetric capnography and a validated equation in the survival prediction of patients with acute respiratory distress syndrome.

PURPOSE: This prospective observational study aims to evaluate the accuracy of dead-space fraction derived from the ventilator volumetric capnography (volumetric CO2) or a prediction equation to predict the survival of mechanically ventilated patients with acute respiratory distress syndrome (ARDS). METHODS: Consecutive VD/VT measurements were obtained based upon a prediction equation validated by Frankenfield et al for dead-space ventilation fraction: VD/VT = 0.320 + 0.0106 (PaCO2-ETCO2)⁺ 0.003 (RR)⁺0.0015 (age) in adult patients who had infection-related severe pneumonia and were confirmed as having ARDS. Here PaCO2 is the arterial partial pressure of carbon dioxide in mmHg; ETCO2, the end- tidal carbon dioxide measurement in mmHg; RR, respiratory rate per minute; and age in years. Once the patient had intubation, positive end expiratory pressure was adjusted and after Phigh reached a steady state, VD/VT was measured and recorded as the data for the first day. VD/VT measurement was repeated on days 2, 3, 4, 5 and 6. Meanwhile we collected dead-space fraction directly from the ventilator volu- metric CO2 and recorded it as Vd/Vt. We analyzed the changes in VD/VT and Vd/Vt over the 6-day period to determine their accuracy in predicting the survival of ARDS patients. RESULTS: Overall, 46 patients with ARDS met the inclusion criteria and 24 of them died. During the first 6 days of intubation, VD/VT was significantly higher in nonsurvivors on day 4 (0.70 ± 0.01 vs 0.57 ± 0.01), day 5 (0.73 ± 0.01 vs. 0.54 ± 0.01), and day 6 (0.73 ± 0.02 vs. 0.54 ± 0.01) (all p =0.000). Vd/Vt showed no significant difference on days 1e4 but it was much higher in nonsurvivors on day 5 (0.45 ± 0.04 vs. 0.41 ± 0.06) and day 6 (0.47 ± 0.05 vs. 0.40 ± 0.03) (both p=0.008). VD/VT on the fourth day was more accurate to predict survival than Vd/Vt. The area under the receiver-operating characteristic curve for VD/VT and Vd/Vt in evaluating ARDS patients survival was day 4 (0.974 ± 0.093 vs. 0.701 ± 0.023, p = 0.0024) with the 95% confidence interval being 0.857-0.999 vs. 0.525-0.841. CONCLUSION: Compared with Vd/Vt derived from ventilator volumetric CO2, VD/VT on day 4 calculated by Frankenfield et al's equation can more accurately predict the survival of ARDS patients.

[Cluster analysis on TCM syndrome characteristics of chronic hepatitis B].

OBJECTIVE: To explore the characteristics of Chinese medicine (CM) syndromes of chronic hepatitis B (CHB) in Sichuan area, thus providing referential evidence for objective research of CHB. METHODS: According to the CRF chart, 1 064 CHB patients' four diagnostic information from 8 clinical units were recorded in Sichuan region using cross-sectional method. The laws of CHB syndrome characteristics were explored using the K-means clustering analysis method. RESULTS: Based on the K-means cluster analysis, we found 8 categories that fulfill the clinical practice combined professional knowledge with experts' opinions. They were Pi-Wei dampness heat (326 cases, 30.6%), Gan and gallbladder dampness heat (193 cases, 18.1%), Gan and Pi dampness heat (158 cases, 14.8%), Gan depression and Pi deficiency (92 cases, 8.6%), Gan depression transforming into heat (89 cases, 8.4%), Pi deficiency with dampness encumbrance (74 cases, 7.0%), dampness-heat combined with yin deficiency of Gan and Shen (73 cases, 6.9%), yang deficiency mingled with blood stasis (59 cases, 5.5%). CONCLUSION: The results of cluster analysis showed Pi-Wei dampness heat, Gan and gallbladder dampness heat, Gan and Pi dampness heat, Gan depression and Pi deficiency, Gan depression transforming into heat, Pi deficiency with dampness encumbrance, dampness-heat combined with yin deficiency of Gan and Shen, yang deficiency mingled with blood stasis were mainly syndromes of CHB patients in Sichuan area.

[Differential expression of microRNA in chronic hepatitis B patients of pi-wei dampness-heat syndrome and of gan depression Pi deficiency syndrome: a primary research].

OBJECTIVE: To explore different microRNA expression profiles between chronic hepatitis B (CHB) patients of Pi-Wei dampness-heat syndrome (PWDHS) and Gan depression Pi deficiency syndrome (GDPDS). METHODS: By applying gene chip technology, blood samples from CHB patients of PWDHS (3 cases), GDPDS (3 cases), and healthy volunteers (3 cases) were withdrawn and microRNA detected. The microRNA was screened and functional analyses performed by using SAS system. RESULTS: Totally 77 microRNAs with differential expression were screened from CHB patients of PWDHS and healthy volunteers, including 60 up-regulated microRNAs and 17 down-regulated microRNAs. Functions of target genes were mainly associated with transcription factors, gas exchange, adverse stimulating, regulation of enzyme activities, developing of the immune system, and the process of actin filaments. Totally 41 microRNAs with differential expression were screened from CHB patients of GDPDS and healthy volunteers, including 32 up-regulated microRNAs and 9 down-regulated microRNAs. Functions of target genes were mainly associated with binding to nucleotide or chromatin, inhibition and activation of transcription, biosynthesis, regulation of metabolic process, regulation of enzyme activities, developing of the immune system, the process of actin filaments, and IL-12. Totally 6 microRNAs with differential expression were screened from CHB patients of PWDHS and CHB patients of GDPDS, including 1 up-regulated microRNA and 5 down-regulated microRNAs. Functions of target genes were mainly associated with transmembrane transport, regulation of transcription factors, metabolism of hormones, developing of the immune system, the process of actin filaments, regulation of metabolic process, response to exterior stimulation, and so on. CONCLUSION: There existed differentially expressed microRNAs (spectrum) between CHB patients of PWDHS and CHB patients of GDPDS.

[A preliminary study of using the expiratory limb of the pressure-volume curve to estimate the dead space in patients suffering from acute respiratory distress syndrome].

OBJECTIVE: To trace pressure-volume curves (P-V curves) with quasi-static technique in acute respiratory distress syndrome (ARDS) patients, and using it to explain the relationship between the end point of the expiratory limb and the change in the dead space of expiration. METHODS: A prospective study was conducted. Fourteen ARDS patients receiving mechanical ventilation were included in the study. When P-V curves were traced with quasi-static technique, the spirometer was connected to the flow sensor. The start point of the expiratory limb was defined as tidal volume 1 (VT1) and the end point as VT2, and the difference between them (ΔVT=VT1-VT2) was calculated. The VT of spirometer (named VT3) and the predetermined VT (VT4) were recorded. Correlations of ΔVT and VT3, as well as VT4 and VT1, were analyzed with correlation analysis. RESULTS: ΔVTwas (417.40±119.68) ml, and VT3 was (399.29±121.36) ml, ΔVT and VT3 showed a good correlation (r=0.99, P=0.000), demonstrating that the ΔVT of the expiratory limb was correlated with the volume trapped in lung. VT4 was (908.21±106.52) ml, and VT1 was(892.26±106.32) ml, and they showed a good correlation (r=0.99, P=0.000). CONCLUSION: Because of part of the gas trapped at the end of expiration in ARDS patients, the dead space is increased, showing that the expiratory limb of the P-V curve cannot return to the base points, and the ΔVT of the expiratory limb is inversely proportional to the gas volume trapped in lung.

[Clinical research of using optimal compliance to determine positive end-expiratory pressure].

OBJECTIVE: To observe the availability and security of optimal compliance strategy to titrate the optimal positive end-expiratory pressure (PEEP), compared with quasi-static pressure-volume curve (P-V curve) traced by low-flow method. METHODS: Fourteen patients received mechanical ventilation with acute respiratory distress syndrome (ARDS) admitted in intensive care unit (ICU) of Tianjin Third Central Hospital from November 2009 to December 2010 were divided into two groups(n = 7). The quasi-static P-V curve method and the optimal compliance titration were used to set the optimal PEEP respectively, repeated 3 times in a row. The optimal PEEP and the consistency of repeated experiments were compared between groups. The hemodynamic parameters, oxygenation index (OI), lung compliance (C), cytokines and pulmonary surfactant-associated protein D (SP-D) concentration in plasma before and 2, 4, and 6 hours after the experiment were observed in each group. RESULTS: (1) There were no significant differences in gender, age and severity of disease between two groups. (2)The optimal PEEP [cm H(2)O, 1 cm H(2)O=0.098 kPa] had no significant difference between quasi-static P-V curve method group and the optimal compliance titration group (11.53 ± 2.07 vs. 10.57 ± 0.87, P>0.05). The consistency of repeated experiments in quasi-static P-V curve method group was poor, the slope of the quasi-static P-V curve in repeated experiments showed downward tendency. The optimal PEEP was increasing in each measure. There was significant difference between the first and the third time (10.00 ± 1.58 vs. 12.80 ± 1.92, P < 0.05). And the optimal compliance titration method had good reproducibility as the optimal PEEP without significant difference in each measure. (3) After the quasi-static P-V curve traced, the heart rate (HR, bpm), temperature (centigrade), interleukin-6 (IL-6, ng/L), tumor necrosis factor-α (TNF-α, ng/L), SP-D (µg/L) showed a gradually increasing tendency, the mean artery pressure (MAP, mm Hg, 1 mm Hg = 0.133 kPa), continuous cardiac index [CCI, L×min(-1)×m(-2)], OI (mm Hg), and C [ml/cm H(2)O] showed a gradually decreased tendency, all of these parameters reached the peak or trough at 6 hours after the experiment, and there was significance compared with those before experiment (HR: 117.34 ± 8.53 vs. 93.71 ± 5.38, temperature: 38.05 ± 0.73 vs. 36.99 ± 1.02, IL-6: 144.84 ± 23.89 vs. 94.73 ± 5.91, TNF-α: 151.46 ± 46.00 vs. 89.86 ± 13.13, SP-D: 33.65 ± 8.66 vs. 16.63 ± 5.61, MAP: 85.47 ± 9.24 vs. 102.43 ± 8.38, CCI: 3.00 ± 0.48 vs. 3.81 ± 0.81, OI: 62.00 ± 21.45 vs. 103.40 ± 37.27, C: 32.10 ± 2.92 vs. 49.57 ± 7.18, all P < 0.05). The results suggested that the drawing of quasi-static P-V curve would aggravate the lung injury. And in optimal compliance titration method group, there were no significant differences in HR, MAP, temperature, CCI, OI, C, cytokines and SP-D before and after titration. CONCLUSION: Optimal compliance titration method has good reproducibility, safety and usability.