Wogonin induces apoptosis in macrophages by exhibiting cytotoxic and genotoxic effects.

Macrophages play an important role in defending the body against invading pathogens. In the face of pathogens, macrophages become activated and release toxic materials that disrupt the pathogens. Macrophage overactivation can lead to severe illness and inflammation. Wogonin has several therapeutic effects, including anti-inflammatory, anticancer, antioxidant, and neuroprotective effects. No studies have investigated the cytotoxic effects of wogonin at concentrations of more than 0.1 mM in RAW264.7 cells. In this study, RAW 264.7 cells were treated with wogonin, which, at concentrations of more than 0.1 mM, had cytotoxic and genotoxic effects in the RAW264.7 cells, leading to apoptosis and necrosis. Further, wogonin at concentrations of more than 0.1 mM induced caspase-3, caspase-8, and caspase-9 activation and mitochondrial dysfunction and death receptor expression. These results suggest that wogonin induces apoptosis through upstream intrinsic and extrinsic pathways by exhibiting cytotoxic and genotoxic effects.

Cyclizine induces cytotoxicity and apoptosis in macrophages through the extrinsic and intrinsic apoptotic pathways.

Cyclizine, an over-the-counter and prescription antihistamine, finds widespread application in the prevention and treatment of motion sickness, encompassing symptoms such as nausea, vomiting, dizziness, along with its effectiveness in managing vertigo. However, the overuse or misuse of cyclizine may lead to hallucinations, confusion, tachycardia, and hypertension. The molecular mechanisms underlying cyclizine-induced cytotoxicity and apoptosis remain unclear. During the 24 h incubation duration, RAW264.7 macrophages were exposed to different concentrations of cyclizine. Cytotoxicity was assessed through the lactate dehydrogenase assay. Flow cytometry employing annexin V-fluorescein isothiocyanate and propidium iodide was utilized to evaluate apoptosis and necrosis. Caspase activity and mitochondrial dysfunction were evaluated through a fluorogenic substrate assay and JC-1 dye, respectively. Flow cytometry employing fluorogenic antibodies was utilized to evaluate the release of cytochrome c and expression of death receptor, including tumor necrosis factor-α receptor and Fas receptor. Western blotting was utilized to evaluate the expression of the Bcl2 and Bad apoptotic regulatory proteins. The findings unveiled from the present study demonstrated that cyclizine exerted a concentration-dependent effect on RAW264.7 macrophages, leading to the induction of cytotoxicity, apoptosis, and necrosis. This compound further activated the intrinsic apoptotic pathway by inducing mitochondrial dysfunction, Bcl2/Bad exchange expression, cytochrome c liberation, and activation of caspases contained caspase 3, 8, and 9. Moreover, the activation of the extrinsic apoptotic pathway was observed as cyclizine induced the upregulation of death receptors and increased caspase activities. Based on our investigations, it can be inferred that cyclizine prompts cytotoxicity and apoptosis in RAW264.7 macrophages in a concentration-dependent manner by triggering both the intrinsic and extrinsic apoptotic pathways.

Critical Care Resources in Guangdong Province of China: Three Surveys from 2005 to 2015.

OBJECTIVES: Data about the critical care resources in China remain scarce. The purpose of this study was to investigate the variation and distribution of critical care resources in Guangdong province from 2005 to 2015. DESIGN: Data in regard to critical care resources were collected through questionnaires and visits every 5 years from 2005. SETTING: All hospitals in Guangdong province were screened and hospitals that provide critical care services were enrolled. INTERVENTION: None. MEASUREMENTS AND MAIN RESULTS: One hundred eleven, 158, and 284 hospitals that provide critical care services were enrolled in the three consecutive surveys respectively. The number of ICUs, ICU beds, intensivists, and nurses increased to 324, 3,956, 2,470, and 7,695, respectively, by 2015. Adjusted by population, the number of ICU beds per 100,000 (100,000) population increased by 147.7% from 2005 to 2015, and the number of intensivists and nurses per 100,000 population increased by 35.3% and 55.1% from 2011 to 2015. However, the numbers in the Pearl River Delta, a richer area, were higher than those in the non-Pearl River Delta area (ICU beds: 4.64 vs 2.58; intensivists: 2.90 vs 1.61; nurses: 9.30 vs 4.71 in 2015). In terms of staff training, only 17.85% of intensivists and 14.29% of nurses have completed a formal accredited critical care training program by 2015. CONCLUSIONS: Our study was the first one to investigate the trend and distribution of critical care resources in China. The quantity of ICU beds and staff has been increasing rapidly, but professional training for staff was inadequate. The distribution of critical care resources was unbalanced. Our study can be beneficial for healthcare policymaking and the allocation of critical care resources in Guangdong province and other provinces in China.

Circulating Neutrophil-Derived Microparticles Associated with the Prognosis of Patients with Sepsis.

INTRODUCTION: Because of its high morbidity and mortality, sepsis remains the leading cause of death in the ICU. Microparticles (MP) have been largely studied as potential diagnostic or prognostic markers in various diseases including sepsis. OBJECTIVE: The biological and clinical relevance of neutrophil-derived microparticles (NDMPs) within the MP population remains unclear. The objective of this study was to elucidate the relationship between plasma NDMPs and the prognosis of patients with sepsis and/or septic shock. METHODS: The study was designed as an observational, noninterventional clinical study. The cohort for this study included 40 sepsis and 40 septic shock patients together with 10 healthy controls admitted to the Intensive Care Unit (ICU) and the Health Surveillance Center in the Affiliated Hospital of Guangdong Medical University, Zhanjiang, Guangdong, China, from January to November 2018, respectively. The degree of critical disease for sepsis and septic shock was evaluated, with data analyses conducted from 2018 to 2019. RESULTS: On days 1, 3 and 5 post-admission a series of data including plasma NDMP levels, patient demographics, TNF-α levels, IL-6 levels, sTREM-1 levels, and the sepsis severity score measurements were collected. A survival curve was plotted against levels of plasma NDMPs. Levels of NDMPs were observed to be higher in the septic shock patients than in the sepsis patients on days 1, 3, and 5 post-ICU admission (p < 0.05). NDMP levels were significantly increased in sepsis and septic shock patients with a parallel increase in pro-inflammatory mediators and sepsis severity score (p < 0.05) as well as mortality. CONCLUSION: Our data suggest that NDMPs may be a biomarker of sepsis severity and mortality although its implications on sepsis prognosis warrant further study.

[Clinical study on effects of ulinastatin on patients with systemic inflammatory response syndrome].

OBJECTIVE: To evaluate the value of ulinastatin in hindering systemic inflammatory response syndrome (SIRS) to proceed to multiple organ dysfunction syndrome (MODS). METHODS: Sixty patients were randomly divided into routine treatment group (n=30) and ulinastatin treatment group (n=30). Both groups were given routine treatment, while the patients of the ulinastatin treatment group were given ulinastatin (100 kU intravenously drip, once every 8 hours, and continued for 5 days) in addition. Additionally, 15 healthy persons were enrolled as normal control group. Temperature (T), heart rate (HR), respiration rate (RR) and white blood cell (WBC) count were observed everyday. The duration of SIRS, the number of organ dysfunction, and mortality were also compared. Serum C reactive protein(CRP), tumor necrosis factor-alpha (TNF-alpha), interleukin-6(IL-6) and IL-10 levels were measured before treatment and 5 days after in ulinastatin treatment group, routine treatment group, and in normal control group at the time of health examination. RESULTS: All of the SIRS markers were not different both in ulinastatin group and routine treatment group before treatment. T, RR, HR and WBC were reduced significantly after 3 days in ulinastatin groups(P<0.05 or P<0.01), but HR was not lowered significantly after 5 days and WBC after 7 days of treatment in regular treatment group (P<0.05 or P<0.01). All of the cytokines in ulinastatin treatment group and routine treatment group were higher than normal control group before treatment. Serum CRP, TNF-alpha and IL-6 levels were reduced significantly after 5 days of treatment in both ulinastatin treatment group and routine treatment group(P<0.01), but in ulinastatin treatment group the reduction was faster than routine treatment group(both P<0.01). IL-10 level was elevated significantly after treatment in ulinastatin treatment group(P<0.01), but it showed no significant change in routine treatment groups(P>0.05). The number of patients with duration of SIRS longer than 3 days were fewer and the incidence of MODS was lower in ulinastatin treatment group than those in routine treatment group(10.00% vs. 36.67%, P<0.05), and the fatality rate was reduced significantly with ulinastatin(3.33% vs. 20.00%, P<0.05). CONCLUSION: Ulinastatin significantly improves the inflammatory symptom and signs of SIRS, such as T, HR, RR, and WBC, inhibits the production of inflammatory cytokines, and enhance the anti-inflammatory cytokines in the treatment of SIRS. It can effectively prevent SIRS to proceed to MODS.

Relationships between genetic polymorphisms of triggering receptor expressed on myeloid cells-1 and septic shock in a Chinese Han population.

BACKGROUND: Triggering receptor expressed on myeloid cells-1 (TREM-1) is a cell surface receptor expressed on neutrophils and monocytes. TREM-1 acts to amplify inflammation and serves as a critical mediator of inflammatory response in the context of sepsis. To date, the predisposition of TREM-1 gene polymorphisms to septic shock has not been reported. This study was designed to investigate whether TREM-1 genomic variations are associated with the development of septic shock. METHODS: We genotyped two TREM-1 single nucleotide polymorphisms (SNPs, rs2234237 and rs2234246) and evaluated the relationships between these SNPs and septic shock on susceptibility and prognosis. RESULTS: TREM-1 rs2234246 A allele in the promoter region was significantly associated with the susceptibility of septic shock in recessive model (AA, OR=3.10, 95%CI 1.15 to 8.32, P=0.02), and in codominant model (AG, OR=0.72, 95%CI 0.43-1.19, P=0.02; AA, OR=2.71, 95%CI 1.00-7.42; P=0.03). However, in three inherited models (dominant model, recessive model, and codominant model), none of the assayed loci was significantly associated with the prognosis of septic shock. The non-survivor group demonstrated higher plasma IL-6 levels (99.7±34.7 pg/mL vs. 61.2±26.5 pg/mL, P<0.01) than the survivor group. Plasma concentrations of IL-6 among the three genotypes of rs2234246 were AA 99.4±48.9 pg/mL, AG 85.4±43 pg/mL, and GG 65.3±30.7 pg/mL (P<0.01). The plasma concentrations of IL-6 in patients with AA genotypes were significantly higher than those in patients with GG genotypes (P<0.01). CONCLUSION: TREM-1 genetic polymorphisms rs2234246 may be significantly correlated only with susceptibility to septic shock in the Chinese Han population.

[Value of C-reactive protein in the diagnosis of arteriosclerosis in patients with essential hypertension].

OBJECTIVE: To evaluate the value of C-reactive protein (CRP) in the diagnosis of arteriosclerosis in patients with essential hypertension. METHODS: This study included 771 patients with essential hypertension and 243 healthy individuals, and all the subjects were measured for carotid-femoral pulse wave velocity (cfPWV) and serum CRP level using enzyme-linked immunosorbent assay (ELISA). The value of CRP in the diagnosis of arteriosclerosis in patients with essential hypertension was evaluated according to the receiver operating characteristics (ROC) curve, and the diagnostic sensitivity and specificity was evaluated with cfPWV> or =9 m/s as the golden diagnostic standard. RESULTS: The hypertensive patients had significantly higher cfPWV and serum CRP concentration than the healthy individuals (16.51-/+1.6 vs 9.81-/+1.1, P<0.001; 4.96-/+1.15 vs 3.52-/+0.33, P<0.001, respectively). CRP showed significant positive correlations to systolic blood pressure (SBP) and pulse pressure (PP) (r=0.584, P<0.001; r=0.624, P<0.001), and when controlled for age, SBP and PP, CRP was found in close correlation to cfPWV (r=0.746, P<0.001). The AUCROC of CRP was 0.907, and the peak point of the ROC curve was 3.85 mg/L, at which point CRP showed a diagnostic sensitivity of 83.9% and specificity of 86.8% with a misdiagnosis rate of 13.2% for arteriosclerosis. CONCLUSIONS: Arteriosclerosis and nonspecific inflammation are prevalent in patients with essential hypertension, and CRP with the cutoff value of 3.85 mg/L may serve as a sensitive indicator for arteriosclerosis diagnosis in these patients.

[Influence of needle-pricking bleeding combined with pulling-rotating manipulation on blood rheology in patients with vertebral artery type cervical spondylosis].

OBJECTIVE: To observe the effect of needle-pricking bleeding combined with pulling-rotating manipulation and simple manipulation on blood rheology in vertebral artery type cervical spondylosis (VATCS) patients, so as to analyze their mechanisms in relieving VATCS. METHODS: A total of 198 VATCS patients were randomly divided into treatment group (n=101) which was treated with needle-pricking plus pulling-rotating manipulation, and control group (n=97) which was treated with simple pulling-rotating manipulation, according to the random number table. The treatment was given once every 7 days, 9 times altogether. The peak systolic blood flow velocity (Vpeak), end-diastolic blood velocity (Vmin), pulsatility index (PI) and resistent index (RI) of bilateral vertebral arteries (VA) and basilar artery (BA) were detected by transcranial doppler sonography (TCD). Whole blood apparent viscosity and the plasma viscosity in the treatment group were determined by using a blood viscosimeter. RESULTS: Of the 101 and 97 VATCS cases in the treatment and control groups, 62 (61.38%) and 12 (12.37%) were cured basically, 23 (22.77%) and 26 (26.80%) experienced marked improvement, 14 (13.86%) and 41 (42.27%) were improved. 2 (1.98%) and 18 (18.55%) failed in the treatment, with the total effective rates being 98.01% and 81.44% separately. The effective rate of the treatment group was significantly higher than that of control group (P<0.05). Compared with pretreatment, Vpeak and Vmin of the bilateral VA and BA in the treatment group, and Vpeak of the right VA in the control group increased significantly (P<0.01, P<0.05), PI and RI of the bilateral VA and BA in the treatment group, and PI and RI of the right VA in the control group decreased significantly (P<0.01, P<0.05), suggesting a marked reduction of the vascular resistance and an apparent increase of the cerebral blood supply after the treatment. The therapeutic effects of the above-mentioned indexes of treatment group were significantly superior to those of the control group (P<0.05). In comparison with pre-treatment, the whole blood apparent viscosity (high, medium and low shear rates) and plasma viscosity of the treatment group post-treatment were obviously reduced (P<0.01, P<0.05). CONCLUSION: Needle-pricking therapy combined with pulling-rotating manipulation can significantly improve VATCS patients' clinical symptoms, which may be closely related to its effects in lowering vascular blood resistance and blood viscosity and increasing cerebral blood supply.

[Variation of serum soluble CD14 levels in patients with chronic heart failure].

OBJECTIVE: To analyze the patterns of serum soluble CD14 (sCD14) and C-reactive protein (CRP) alterations in patients with chronic heart failure (CHF) and investigate the correlations of sCD14 variation to the etiology, clinical symptoms, and the number of mononuclear cells in these patients. METHODS: This study involved 246 CHF patients stratified according to their etiology and clinical symptoms, with 107 normal individuals serving as the control group. Blood samples were collected from these patients the next day after admission and also from the control subjects for measuring serum sCD14 and CPR levels using enzyme-linked immunosorbent assay (ELISA) and rate nephelometry, respectively. RESULTS: The CHF patients had significantly higher serum levels of sCD14 and CRP than the control subjects (P<0.01). In the CHF patients, serum sCD14 and CRP levels differed significantly in the patients with clinical symptoms of different severities (F=3.787, P=0.024), and those with moderate and severe symptoms had significantly higher levels than the asymptomatic patients (P<0.05). The difference in etiologies also resulted in significant difference in sCD14 levels (P<0.05), which were significantly lower in coronary artery disease group than in hypertension group (P<0.05). Significant positive correlations were found between sCD14 and the CRP levels in the CHF patients (r=0.227, P=0.018) and between sCD14 level and the clinical symptoms (r=0.206, P=0.001), but sCD14 level was not correlated to the absolute or relative number of mononuclear cells. CONCLUSIONS: Serum sCD14 and CRP levels are significantly elevated in CHF patients, but this condition may vary as the etiologies and clinical symptoms differ. Increased mononuclear cells do not contribute to the elevation of serum sCD14.

[Mechanisms of losartan for inhibition of myocardial fibrosis following myocardial infarction in rats].

OBJECTIVE: To investigate the effect of losartan on cardiac mineralocorticoid receptor (MR) mRNA in rats after acute myocardial infarction (AMI). METHOD: AMI was induced in male SD rats by ligation of the left coronary artery. The survived rats were randomly divided into AMI group, losartan group, and sham-operated group. The cardiac functions of the rats were assessed by echocardiogram and hemodynamics, and the contents of angiotensin II (Ang II) and aldosterone (Ald) in the myocardial tissues were determined by radioimmunoassay. The collagen density in the myocardial tissues were calculated by Masson's trichrome staining and the expression of MR mRNA were determined by real-time quantitative fluorescent PCR. RESULTS: Both the contents of AngII and Ald in the myocardial tissues increased significantly in AMI group compared with those in the sham-operated group (P<0.01). The expression of MR mRNA and collagen density in the myocardial tissues also increased significantly than that in sham-operated group (P<0.01). After four weeks of losartan treatment, the contents of AngII and Ald in the myocardial tissues decreased significantly (P<0.05) and the expression of MR mRNA was also considerably lowered (P<0.01) in comparison with those in the AMI group. Treatment with losartan also resulted in significant decrease of the collagen density in the myocardial tissues. CONCLUSIONS: Losartan may reduce reactive fibrosis not only by attenuating the Ald signaling pathway but also by decreasing the expression of MR.

[Clinical observation and study of mechanisms of needle-picking therapy for primary infertility of abnormal sperm].

OBJECTIVE: To search for the best method for increasing clinical therapeutic effect on primary abnormality of sperm. METHODS: One hundred and sixty-eight cases of infertility were randomly divided into a treatment group of 85 cases and a control group of 83 cases. The treatment group were treated with needle-picking at bilateral Shengzhi points, Dicong Shenjing points and L2 Shenjing points as main. The control group were treated with oral administration of Wuzi Yanzong Pills [symbol: see text]. Their therapeutic effects were observed in 3 hospitals. RESULTS: The total effective rate of 83.5% and the pregnancy rate of the patient's wife of 78.8% in the treatment group, and corresponding figures were 54.2% and 43.4% in the control group, with a very significant difference between the two groups (P < 0.01); and reproductive hormones improved significantly after treatment in the two groups (P < 0.01); after treatment, superoxide dismulase (SOD) activity and Zn content in semen were elevated and cadmium level decreased significantly in the treatment group (P < 0.05). CONCLUSION: Needle picking therapy can significantly improve and regulate endocrine function, increase quality of semen and elevate pregnancy rate of the patient's wife for the patient of primary abnormal sperm.