Identification of key genes related to immune infiltration in cirrhosis via bioinformatics analysis.

Cirrhosis is the most common subclass of liver disease worldwide and correlated to immune infiltration. However, the immune-related molecular mechanism underlying cirrhosis remains obscure. Two gene expression profiles GSE89377 and GSE139602 were investigated to identify differentially expressed genes (DEGs) related to cirrhosis. Enrichment analysis for DEGs was conducted. Next, the immune infiltration of DEGs was evaluated using CIBERSORT algorithm. The hub DEGs with tight connectivity were identified using the String and Cytoscape databases, and the expression difference of these hub genes between normal liver and cirrhosis samples was determined. Moreover, in order to evaluate the discriminatory ability of hub genes and obtained the area under the receiver operating characteristic curve values in the GSE89377 and GSE139602 datasets. Finally, the association between hub DEGs and immune cell infiltration was explored by Spearman method. Among the 299 DEGs attained, 136 were up-regulated and 163 were down-regulated. Then the enrichment function analysis of DEGs and CIBERSORT algorithm showed significant enrichment in immune and inflammatory responses. And four hub DEGs (ACTB, TAGLN, VIM, SOX9) were identified, which also showed a diagnostic value in the GSE89377 and GSE 139,602 datasets. Finally, the immune infiltration analysis indicated that, these hub DEGs were highly related to immune cells. This study revealed key DEGs involved in inflammatory immune responses of cirrhosis, which could be used as biomarkers for diagnosis or therapeutic targets of cirrhosis.

Analysis on clinical features of 17 death cases of imported malaria in China / 中国热带医学

@#Abstract: Objective　China was certified by World Health Organization as a malaria-free country in 2021. Malaria has become a rare infectious disease, and preventing the re-transmission of imported malaria and reducing deaths are the main challenges facing China after elimination of malaria. To analyze and clarify the characteristics of imported malaria deaths, and to provide prevention and treatment recommendations for overseas workers and health care workers. Methods　The data of 17 imported malaria deaths in the analysis of malaria deaths from 2016 to 2020 by the National Severe Malaria Treatment Expert Group were collected, and the relevant clinical epidemiological data and disease course records were analyzed. Results　The 17 malaria deaths were all imported from Africa with Plasmodium falciparum infection (malarial cerebral type), with no obvious regularity in the month of onset. Among them, 16 were male patients, 5 cases with underlying diseases such as diabetes mellitus, and 10 patients were first diagnosed in a second-level or lower hospital. Excluding patients who died of respiratory cardiac arrest in ambulances, the mean time difference between first onset and malaria diagnosis in 16 patients was 6.8 days (median 5.5 days), and the mean time between first onset and antimalarial treatment was 7.4 days (median 6 days), the mean time difference from initial onset to death was 10.3 days (median 8.5 days). Excluding cases with onset abroad and unknown time of return, all 14 patients developed the disease within 30 days after returning to China. Conclusion　All the fatal cases were infected with Plasmodium falciparum imported from Africa. The patients' awareness of actively seeking medical treatment is weak, and the delay in seeking medical treatment caused by the insufficient diagnosis and treatment capacity of health institutions at the township level and below is the main reason for the deaths. It is recommended to strengthen the self-protection awareness of staff in malaria-endemic areas overseas and raise their awareness of malaria. For returnees from areas with high malaria risk, primary medical institutions should pay attention to the patient's travel history in Africa, improve the awareness of malaria diagnosis, malaria diagnosis and treatment capabilities.

Nicotine ameliorates experimental severe acute pancreatitis via enhancing immunoregulation of CD4+ CD25+ regulatory T cells.

OBJECTIVES: Activation of "nicotinic anti-inflammatory pathway" could reduce severity of inflammation and injury induced by acute pancreatitis. However, the role of regulatory T (Treg) cells in this pathway is unclear. METHODS: Severe acute pancreatitis (SAP) was induced in mice through retrograde injection of 50-µL 2% Na-taurocholate into the pancreatic duct of the mouse. In nicotine treatment group, nicotine (50, 100, and 300 µg/kg) was administered 1 hour before and after SAP operation through intraperitoneal injection. We compared the properties of Treg cell percentage and specific marker such as cytotoxic T-lymphocyte antigen 4 and forkhead box transcription factor forkhead/winged helix transcription factor p3 on Treg using quantitative reverse transcription polymerase chain reaction and flow cytometry. All experiment animal serum cytokines were measured using enzyme-linked immunosorbent assay. One-way analysis of variance was applied to evaluate the experimental data and for statistical comparisons. The survival rate data were analyzed using the log-rank test. RESULTS: Nicotine significantly protected mice from lethal SAP in a dose-dependent fashion by inhibiting tissue injury, digestive enzyme production, and proinflammatory cytokines production. Moreover, nicotine up-regulated the number and suppressive capacity of CD4 CD25 Treg via inducing the expression of immunoregulatory molecules and transforming growth factor ß1 elevation. CONCLUSIONS: Modulating immunoregulation of CD4 CD25 Treg is a critical mechanism for nicotinic anti-inflammatory pathway and it may be feasible to use selective agonists as an immunotherapy for SAP.

Codonopsis pilosula polysaccharide attenuates cecal ligation and puncture sepsis via circuiting regulatory T cells in mice.

BACKGROUND: Codonopsis pilosula polysaccharide (CPPS) isolated from one of the Chinese herbs is known to have a variety of immunomodulatory activities. However, it is not clear whether CPPS can exert an effect on the immune functions of regulatory T cells (Tregs). This study was carried out to investigate the effect of CPPS on the immune function of peripheral blood Tregs in sepsis induced by cecal ligation and puncture (CLP). METHODOLOGY AND PRINCIPAL FINDINGS: BALB/c mice were randomly divided into five groups: sham group, CLP group, CLP with CPPS (40, 100, and 250 mg/kg) treatment group, and they were killed on days 1, 2, 3, and 4 after CLP, respectively, with eight animals at each time point. Magnetic microbeads were used to isolate peripheral blood Tregs and CD4 T cells. Phenotypes of Tregs, such as Toll-like receptor 4 (TLR4) and Foxp3, were analyzed by flow cytometry, and coculture medium cytokines levels were determined with enzyme-linked immunosorbent assay. The levels of TLR4 and the expression of Foxp3 in the Treg from CLP group were markedly increased in comparison to the sham group. Administration of CPPS could significantly decrease the TLR4 level and inhibited the expression of Foxp3 on Tregs in sepsis mice. At the same time, proliferative activity and expression of interleukin 2 and interleukin 2Rα on CD4 T cells were restored. In contrast, anti-TLR4 antibody could block the effect of CPPS on Treg immune function. CONCLUSIONS: Codonopsis pilosula polysaccharide might suppress excessive Tregs, at least in part, via TLR4 signaling on Tregs and trigger a shift of TH2 to TH1 with activation of CD4 T cells in sepsis induced by CLP.

Expression of triggering receptor-1 in myeloid cells of mice with acute lung injury.

BACKGROUND: Myeloid cell (TREM-1) is an important mediator of the signal transduction pathway in inflammatory response. In this study, a mouse model of acute lung injury (ALI) by intraperitoneal injection of lipopolysaccharide (LPS) was established to observe the expression pattern of TREM-1 in lung tissue and the role of TREM-1 in pulmonary inflammatory response to ALI. METHODS: Thirty BALB/C mice were randomly divided into a normal control group (n=6) and an ALI group (n=24). The model of ALI was made by intraperitonal injection of LPS in dose of 10 mg/kg. Specimens from peripheral blood and lung tissue were collected 6, 12, 24 and 48 hours after LPS injection. RT-PCR was used to detect TREM-1 mRNA, and ELISA was employed for detection of TREM-1 protein and TNF-a protein, and HE staining was performed for the pathological Smith lung scoring under a light microscope. RESULTS: The expressions of TREM-1 mRNA in lung tissue and blood of the ALI group 6, 12, 24, and 48 hours after injection of LPS were higher than those in the control group. The levels of TREM-1 protein and the levels of TNF-a protein in lung tissue of the ALI group 6, 12, 24, and 48 hours after LPS injection were higher than those of the control group; the level of TREM-1 protein peaked 12 hours after LPS injection, but it was not significantly correlated with the expression of TREM-1 mRNA (P=0.14); the TNF-a concentration was positively correlated with TREM-1 levels in lung tissue and with Smith pathological score (r=0.795, P=0.001:r=0.499, P=0.034), but not with the expression of TREM-1 mRNA (P=0.176). CONCLUSIONS: The expression of TREM-1 mRNA in lung tissue of mice with ALI is elevated, and the expression of TREM-1 mRNA is related to the level of TNF-a and the severity of inflammatory response to ALI. The expressions of the TREM-1 gene are not consistent with the levels of TREM-1 protein, suggesting a new functional protein involved in immune regulation.

[The effect of blood glucose fluctuations on prognosis of critically ill patients in intensive care unit].

OBJECTIVE: To determine the association between glucose fluctuations and hospital mortality in intensive care unit (ICU) patients. METHODS: A retrospective study involving 90 critically ill patients in ICU according to the patients' outcome were divided into survivors (49 cases) and nonsurvivors (41 cases), in whom the blood glucose level was monitored in the first 72 hours, and the initial blood glucose (GluAdm), the average blood glucose (GluAve), glucose standard deviation (GluSD), coefficient of variation glucose (GluCV) were determined, then GluAdm, GluAve, GluSD, and GluCV were compared between survivors and nonsurvivors, and the receiver operating characteristic curve (ROC curve) was applied to evaluate the association between blood glucose fluctuation and prognosis. According to the values of GluSD, GluCV, the critically patients were divided into four subgroups, and mortality in ICU and hospital was compared. RESULTS: The levels of GluAdm, GluAve, GluSD, GluCV of nonsurvivors were higher than those of survivors [GluAdm: (11.47+/-3.91) mmol/L vs. (9.23+/-2.96) mmol/L, GluAve: (9.22+/-1.31) mmol/L vs. (8.28+/-1.15) mmol/L, GluSD: (2.62+/-0.97) mmol/L vs. (1.66+/-0.64) mmol/L, GluCV: 0.28+/-0.10 vs. 0.20+/-0.07, all P<0.05]. When ROC was applied, the area under the curve (AUC) of GluSD, GluCV were 0.782+/-0.049 and 0.757+/-0.053, they were higher than that of the GluAdm and GluAve (0.669+/-0.058 and 0.690+/-0.056, both P<0.05). When GluSD was 4.35-5.66 mmol/L, the ICU mortality was 95.7%, hospital mortality was 98.6%; when GluCV was 0.378-0.500, the ICU mortality was 83.3%, hospital mortality was 100.0%. CONCLUSION: The increase in GluSD and GluCV in critically ill patients is significantly correlated with ICU mortality and hospital mortality, and they are more valuable in predicting ICU mortality than GluAdm, GluAve. Diminution in fluctuation of blood glucose might be an important aspect of glucose management.

[The long term effect of small dose budesonide turbuhaler on mild bronchial asthma].

OBJECTIVE: To investigate the long term efficacy as well as the side-effects of small dose budesonide turbuhaler in mild bronchial asthma. METHODS: Fifty-two patients with mild asthma were randomly divided into groups A, B and C. Twenty-two cases in group A received 200 microg budesonide turbuhaler by inhalation every night. Fifteen cases in group B received oral prednisone 5 mg/d and theophyllin control release tablet 0.2 g twice daily, as well as ventolin aerosol inhalation 200 microg three times daily during asthmatic attack. Fifteen cases in group C received no glucocorticoids, but theophyllin control release tablet 0.2 g twice daily as well as ventolin aerosol inhalation (200 microg/d) were given during asthmatic attack. All three groups received treatment consecutively for 3 years, and followed for one year after stopping treatment. During the total 4 years, pulmonary function measurements (FEV1, Raw, Gaw), bronchial hyperreactivity (BHR), clinical effects, plasma cortisol level and the activated reaction of ACTH were evaluated. RESULTS: (1) Before the treatment, BHR of the three groups were similar, with most cases graded III-IV, being 91% (20/22), 100% (15/15) and 93% (14/15) respectively. After the treatment, the BHR of 18 (82%) cases of group A was reduced to grades I-II. However, 13 cases (87%) of group B still remained at grades III-IV and 15 cases (100%) of group C were still at grades III-IV. The results of the three groups showed a significant difference (P < 0.01). (2) Compared to measurements before the treatment, Raw was (623 +/- 103)% vs (158 +/- 24)% in group A, (605 +/- 90)% vs (340 +/- 61)% in group B, and (638 +/- 108)% vs (420 +/- 81)% in group C, the difference among the three groups being significant (P < 0.01). (3) Compared to the measurements before the treatment, Gaw was (22 +/- 4)% vs (83 +/- 15)% in group A, (27 +/- 6)% vs (42 +/- 9)% in group B, and (27 +/- 5)% vs (31 +/- 6)% in group C, the difference among the three groups being significant (all P < 0.01 respectively). (4) Compared with the measurements before the treatment, FEV1 was (2.3 +/- 0.4) L vs (2.9 +/- 0.4) L in group A, (2.3 +/- 0.4) L vs (2.6 +/- 0.4) L in group B, and (2.3 +/- 0.4) L vs (2.7 +/- 0.4) L in group C, the difference among the three groups being significant (P < 0.01). (5) With asthmatic symptom control as the criteria of therapeutical effects, the effective rate of group A, B and C were 91%, 53% and 33% respectively, which showed significant difference among the three groups (all P < 0.01 respectively). (6) The plasma cortisol levels of group A and B were (326 +/- 103) nmol/L and (308 +/- 29) nmol/L before treatment, compared with (318 +/- 78) nmol/L and (299 +/- 98) nmol/L after treatment, the difference in group A or in group B was not significant (all P > 0.05). When the measurements before and after treatment were compared, the cortisol levels of group A and B were (365 +/- 102) nmol/L vs (373 +/- 102) nmol/L and (343 +/- 79) nmol/L vs (346 +/- 103) nmol/L respectively after the stimulation of ACTH, which showed no significant changes even after the stimulation of ACTH (all P > 0.05). CONCLUSIONS: Long term small dose budesonide turbuhaler inhalation could effectively decrease BHR and the Raw, while increase the Gaw of asthmatic patients, hence improving the pulmonary function and preventing acute attack. In addition, it did not induce suppression of HPAA axis function in the patients.