Objective To investigate the effects of 5-Aza-CdR( methylation transferase inhibitor) on the expression levels of leptin gene in chondrocytes and methylation states of leptin promoter region between osteoarthritis (OA) group and control. Methods The chondrocytes in osteoarthritis group were treated with 5-Aza-CdR with different doses and time-points, and the expression level of leptin was detected by real-time polymerase chain reaction for picking up the optimum dose and time-point. Next, the chondrocytes in 5 osteoarthritis patients and 5 control patients (amputation due to severe trauma) were treated with 5-Aza-CdR. Lastly, leptin mRNA expression levels in the four groups osteoarthritis and control chondrocytes treated with/without 5-Aza-CdR were measured by real-time PCR and the methylation state of promoter region ( - 280- + 79) was detected by epityper quantitative DNA methylation analysis. Results ( 1 ) After treating the chondrocytes in OA groups with 10 μmol/L 5-Aza-CdR for 72 h, the mRNA expression levels of leptin were increased significantly. ( 2 ) The mRNA expression levels of leptin were significantly different among the four groups ( P ＜ 0. 05 ), and the chondrocytes in osteoarthritis groups treated with 5-Aza-CdR showed a marked induction of leptin mRNA expression. (3) Analysis of quantitative methylation data using an unsupervised hierarchical clustering algorithm, showed that methylation patterns of leptin promoter was different between control and osteoarthritis chondrocyte treated with/without 5-Aza-CdR. Conclusion Demethylation of leptin promoter might up-regulate leptin gene expression level and it might contribute to osteoarthritis.

Objective To investigate the expression of procalcitonin (PCT) and C-reactive protein (CRP),and endotoxin in bloodstream infection with different microorganisms,so as to assess the value of these inflammatory cytokines in early diagnosis of sepsis in bloodstream infections patients.Methods Data of 152 septic bloodstream infected patients with 90 male and 62 female aged from 62 to 102 years and 79.2 ± 16.3 years in average admitted from January 2012 to December 2013 were analyzed retrospectively.According to the results of blood culture,the microorganisms could be categorized into gram-negative bacteria,gram-positive bacteria and fungus groups,and the levels of serum CRP,PCT,and endotoxin were compared among these groups of bloodstream infections patients within 24 hours after admission.Results (1) A total of 152 strains of microorganisms were surveyed including 92 gram-negative strains (61.18％),43 gram-positive strains (28.29％),and 16 fungal strains (10.53％).In the gram-negative strains,Klebsiella pneumoniae (n =29),Acinetobacter baumannii (n =24),Escherichia coli (n =23),Burkholderia cepacia (n =9) and Pseudomonas aeruginosa (n =4) were the most common isolates.In the Gram-positive strains,13 strains of Staphylococcus aureus were isolated.(2) In the gram-negative bacterial bloodstream infections group,there were 60 (64.52％) patients with endotoxin positive,and there were no endotoxin positive cases with detected gram-positive bacteria and fungal bloodstream infections.The median levels of PCT were significantly different among the three groups [gram-negative strains group:7.760 (3.365,28.585) ng/mL,gram-positive strains group:0.705 (0.265,3.225) ng/mL,fungal infection group:1.245 (0.543,1.998) ng/mL].In the fungal bloodstream infection group,the mean level of CRP was higher than that in other two groups [gram-negative strains group:(126.01 ± 66.53) mg/L,gram-positive strains group:(77.58 ±54.21) mg/L,fungal infection group:(140.14 ±71.21) mg/L].(3) The receiver operating characteristic (ROC) curve of inflammatory cytokines was made for the diagnostic value in bloodstream infections.ROC curve used to distinguish between gram-positive bacterial bloodstream infections group and fungal group showed that AUCPCT+CRP =0.791.When PCT cut-off value was 0.92 ng/mL,and CRP cut-off value was 68.00 mg/L,the sensitivity was 50％ and the specificity was 95.5％.ROC curve used to distinguish between the gram-negative bacterial bloodstream infections group and fungal group showed that AUCPCT+CRP+LPS =0.947.When PCT cut-off value was 2.16 ng/mL and CRP cutoff value was 94.10 mg/L,and endotoxin was positive,the sensitivity was 82.8％ and the specificity was 100％.ROC curve used to distinguish between gram-negative bacterial bloodstream infections group and gram-positive bacterial group showed that AUCPCT+CRP+LPS =0.947.When PCT cut-off value was 2.68 ng/ mL,CRP cut-off value was 106.5 mg/L,endotoxin was positive,the sensitivity was 74.2％ and the specificity was 97.7％.Conclusions Gram-negative bacteria were the most common microorganisms in bloodstream infections in ICU patients.Compared with single inflammatory cytokine,the serum concentrations of PCT,CRP and endotoxin used together could provide more sensitivity and specificity for the early diagnosis of bloodstream infection with different microorganisms.

ObjectiveTo investigate the gene expression profiling of articular chondrocyte and the function and pathway of the differentially expressed genes in patients with osteoarthritis (OA) by using gene microarray technique.MethodsThree patients with OA,three patients with rheumatoid arthritis(RA) and three traumatic controls without arthritis were selected and were divided into OA versus RA and OA versus the traumatic control groups,and their articular cartilage cells were cultivated.The gene expression profiling was performed by human genome oligonucleotide microarray technique.The differences of gene expression of the articular chondrocyte in OA versus RA group and OA versus the traumatic control group were compared respectively by two class unpaired test using significant analysis of microarray software.The function and pathway of these differentially expressed genes were analyzed by using the database of Molecule Annotation System.ResultsThe number of differentially expressed genes was 145 when OA compared with the traumatic control,in which 70 were up-regulated and 75 were down-regulated; and the number was 281 when OA were compared with RA,in which 94 were up-regnlated and 187 were down-regulated.The Gene Onto-logy (GO) functions of the differentially expressed genes of OA in each group were related to pathological and immune courses including cellular process,physiological process,cell division,biological regulation and cell signal transduction.The statistically significant pathways of these genes in each group included apoptosis pathway,cell cycle pathway,P53 signaling pathway,Fas signaling pathway,NO pathway,apeptotic cascade pathway,focal adhesion pathway, ECM-receptor interaction pathway, tight junction pathway, adhesive bonding pathway,actin cytoskeleton regulation pathway,bone remodeling pathway,chondroitin sulfate biosynthesis pathway,Jak-STAT signaling pathway,Wnt signaling pathway,Toll recepor signaling pathway,cell adhesion molecules pathway,T cell receptor signaling pathway,and s o on (P＜0.05).They displayed not only marked dif-ferences in GO function and gene pathway of differentially expressed genes between OA versus RA group and OA versus the traumatic control group,but also displayed the significant overlapping of the differentially expressed genes and pathways between the two groups.ConclusionThe differentially expressed genes and pathways of articular chondrocytes involve apoptosis,extracellular matrix and cytokines in OA,which contri-bute to further study of early warning genes of OA.

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Intravenous and intratracheal implantation of mesenchymal stem cells (MSCs) may offer ameliorating effects on pulmonary hypertension (PH) induced by monocrotaline (MCT) in rats. The aim of this study was to examine the anti-remodeling effect of intravenous MSCs (VMSCs) and intratracheal MSCs (TMSCs) in rats with PH, and the underlying mechanisms. MSCs were isolated from rat bone marrow and cultured. PH was induced in rats by intraperitoneal injection of MCT. One week after MCT administration, the rats were divided into 3 groups in terms of different treatments: VMSCs group (intravenous injection of MSCs), TMSCs group (intratracheal injection of MSCs), PH group (no treatment given). Those receiving saline instead of MCT served as negative control (control group). Pulmonary arterial structure was pathologically observed, pulmonary arterial dynamics measured, and remodeling-associated cytokines Smad2 and Smad3 detected in the lungs, three weeks after MCT injection. The results showed that PH group versus control group had higher pulmonary arterial pressure (PAP) and wall thickness index (WTI) 21 days after MCT treatment. The expression of phosphorylated (p)-Smad2 and the ratio of p-Smad2/Smad2 were much higher in PH group than in control group. Fluorescence-labeled MSCs were extensively distributed in rats' lungs in VMSCs and TMSCs groups 3 and 14 days after transplantation, but not found in the media of the pulmonary artery. WTI and PAP were significantly lower in both VMSCs and TMSCs groups than in PH group three weeks after MCT injection. The p-Smad2 expression and the ratio of p-Smad2/Smad2 were obviously reduced in VMSCs and TMSCs groups as compared with those in PH group. In conclusion, both intravenous and intratracheal transplantation of MSCs can attenuate PAP and pulmonary artery remodeling in MCT-induced PH rats, which may be associated with the early suppression of Smad2 phosphorylation via paracrine pathways.

ObjectiveTo observe the role of Sijunzi decoction on the gut barrier protection and immunity regulation in spleen-qi deficiency type patients with persistent inflammation-immunosuppression catabolism syndrome (PICS).Methods A prospective study was conducted, and according to random number table, 46 patients with PICS accompanied by spleen-qi deficiency admitted to Department of Critical Care Medicine of Beijing Shijitan Hospital of Capital Medical University were randomly divided into two groups: control group and Sijunzi decoction group(each 23 cases). Conventional therapy was given to both groups, and the patients in Sijunzi decoction group were additionally treated with modified Sijunzi decoction 100 mL by nasal feeding, while those in the control group were treated with an equal amount of warm boiled water by nasal feeding. The course of treatment was 2 weeks in both groups. The improvement in traditional Chinese medicine(TCM) syndrome and gastrointestinal function was observed in two groups before and after treatment. At the same time, the changes of lymphocyte subsets including total T cells(CD3+ cells),helper/inducer T cells(CD3+/CD4+T cells), suppressor/cytotoxic T cells(CD3+/CD8+ T cells), CD4/CD8, total natural killer cells(NK cell,CD3-/CD16+CD56+ cells),natural killer like T cells(NK T cell,CD3+/CD16+CD56+ T cells),total B cells(CD19+ cells),regulatory T cells(Treg T cells, CD4+/CD25+ T cells),suppressor T cells(CD8+/CD28- T cells) and cytotoxic T cells(CD8+/CD28+ T cells)were analyzed.Results① There were no differences in the acute physiology and chronic health evaluationⅡ(APACHEⅡ)score,acute gastrointestinal injury(AGI)grades and peripheral blood lymphocyte subsets between Sijunzi decoction group and control group before treatment( allP>0.05).② After 2 weeks of treatment, all the patients' clinical symptoms of spleen-qi deficiencywere improved, and there was no difference in effect between the Sijunzi decoction and control group(valid:11 cases vs. 6 cases, invalid:12 cases vs.17 cases, bothP>0.05) . In both groups, the patients' AGI condition was improved, but the improvement in treatment group was more significant than that in the control group(valid: 14 cases vs. 7 cases, P<0.05) .③ In the control group,the numbers of CD3+, CD3+/CD8+,CD8+/CD28- T cells after treatment were increased significantly〔CD3+:(62.37±7.83)% vs.(54.08±11.65)%, CD3+/CD8+:(31.52±10.55)% vs. (23.94±9.22)%, CD8+/CD28-:(24.97±10.25)% vs.(16.78±10.55)%〕 and CD19+ number was decreased obviously〔(5.78±5.33)% vs.(9.73±8.02)%〕 at the same time(allP<0.05). After treatment, in the Sijunzi decoction group, the numbers of CD3+,CD3+/CD4+, CD3+/CD8+, CD8+/CD28-, CD8+/CD28+ T cells were increased significantly compared with those before treatment〔CD3+:(74.53±7.64)% vs.(52.98±10.05)%, CD3+/CD4+:(36.27±12.08)% vs.(30.00±8.60)%, CD3+/CD8+:(37.33±12.56)% vs.(22.88±9.97)%, CD8+/CD28-:(26.89±10.80)% vs.(17.01±9.48)%, CD8+/CD28+:(12.08±5.50)% vs.(8.47±4.29)%〕, and total CD19+ number was decreased remarkably at the same time〔(4.60±4.28)% vs.(9.86±8.61)%,P<0.05〕. In the Sijunzi decoction group, after 2 weeks of treatment, the number of CD3+ T cells was increased more significantly than that in the control group〔(8.29±9.28)% vs.(5.80±5.33)%,P<0.05〕.ConclusionUsing Sijunzi decoction combined with conventional therapy can improve the clinical symptoms of patients with PICS accompanied by spleen-qi deficiency and gastrointestinal dysfunction, the mechanism is possibly via enhancing immunity by regulating the numbers of B and T lymphocytes, and maybe the cell-mediated immunity plays a more prominent role.

Objective To evaluate the effect of combination therapy with milrinone and esmolol on hemodynamics and cardiac function in patients with septic myocardial depression. Methods From October 2010 to October 2013,after the hemodynamics and cardiac function were evaluated by pulse indicator continuous cardiac output (PICCO),74 sepsispatients withCI < 2.2 L/min · m2 after fluid resuscitation were enrolled in the study and were divided into group A with intravenous injection of dobutamine hydrochloride ,and group B with intravenous injection of milrinone and esmolol,with 37 cases in each group. The patients'PICCO indicators, echocardiography and cardiac biomarker(CK,CK-MB,MYO,cTnI and ProBNP)in two groups were compared before and after 3-day treatment. Results (1)CI and GEF were significantly increased in group B after 3-day treatment when compared with those in group A.(2)Compared with those in group A,early diastolic mitral flow velocity/end diastolic mitral velocity (E/A) and right ventricular diastolic diameter(RVD) in group B had statistical significance.(3) CK-MB,cTnI and ProBNP decreased significantly in group B when compared with those in group A. Conclusion Combination therapy with milrinone and esmolol can increase cardiac ejection function,slow down the heart rate,reduce the heart blood and vascular preload,lessen the injury of myocardial and improve heart function.

Objective To investigate the effects of volatile oil from artemisia dracunculus on myocardial injury caused by viral myocarditis in mice and explore its possible mechanism.Methods Totally 160 adult male BALB/c mice were randomly divided into normal control group (10) and viral myocarditis group (150).Viral myocarditis mice models were reproduced by intraperitoneal inoculation with a solution of coxsackievirus B3 (CVB3),a viral strain with affinity to myocardium,and then randomly divided into model,astragalus group,and low-,medium-,and high-dose volatile oil from artemisia dracunculus groups.After 1 hour of viral infection,normal control group and model group mice were given normal saline by intragastric administration,astragalus group mice were injected with astragalus 0.1 mL in each mouse by intraperitoneal injection,and the mice in other three groups were given low,medium and high dose (2％,5％,10％) 0.3 mL volatile oil from artemisia dracunculus in each mouse by intragastric administration,respectively,once a day for one week consecutively.The mortality,heart/body weight ratio,the activity of natural killer cells (NK cell),virus titer in myocardial homogenate,serum cardiac troponin Ⅰ (cTnI) level and myocardial pathological changes were observed.Results ① Mortality:the mortality of model group was higher than that of the normal control group,astragalus group,low and medium dose volatile oil from artemisia dracunculus groups (60.0％ vs.0％,23.3％,20.0％,28.7％),and the difference in the mortality being of no statistical significance between model group and that of high-dose volatile oil from artemisia dracunculus group (60.0％ vs.47.6％,P ＞ 0.05);the mortality of astragalus group was obviously lower than that of high-dose volatile oil from artemisia dracunculus group (P ＜ 0.01),and the differences in comparisons between the mortalities of astragalus intervention group,and medium-and low-dose volatile oil groups were not statistically significant (all P ＞ 0.05),and the comparison of mortality between low-and medium-dose volatile oil groups were also not statistically significant (P ＞ 0.05).② Immunization parameters:on the 8th day after modeling,the activity of NK cells in the model group was significantly lower than that in the normal control group [(15.91 ± 3.87)％ vs.(38.50 ± 2.32)％],the activities of NK cells in astragalus group,medium-and low-dose volatile oil from artemisia dracunculus groups were significantly higher than that in model group [(19.38 ± 3.27)％,(18.54 ± 3.09)％,(18.36 ± 2.64)％ vs.(15.91 ± 3.87)％,all P ＜ 0.05].None of virus was detected in the myocardial homogenate in the normal control group,and the virus titers in astragalus group,low and medium dose volatile oil from artemisia dracunculus groups were significantly lower than the titer of the model group (10-9/mL:1.96 ± 0.44,1.95 ± 0.46,1.95 ± 0.48 vs.2.41 ± 0.51,all P ＜0.01).③ Myocardial injury parameters:the level of cTnI in the normal control group was less than 0.1 μg/L,obviously lower than that in the model group [(15.84 ± 3.89) μg/L],as well as the ratio of heart/body weight in model group was also significantly higher than that in normal control group (× 10-4:8.3 ± 1.3 vs.4.6 ± 0.1),and the cTnI and the ratio of heart/body weight of astragalus intervention group,low and medium dose volatile oil from artemisia dracunculus groups were markedly lower than those of model group [cTnI (mg/L):10.03 ± 2.35,10.81 ± 2.56,11.10 ± 1.89 vs.15.84 ± 3.89,ratio of heart/body weight (× 10-4):7.2 ± 0.8,7.3 ± 1.0,7.3 ± 0.6 vs.8.3 ± 1.3].In the normal control group,there were no inflammatory cell infiltration and necrosis in myocardial tissue,the scores of myocardial pathological changes were 0.In the model group,the scores of inflammatory cell infiltration (3.25 ± 0.45) and of necrosis (2.91 ± 0.51) were markedly higher than those in the normal control group.And the above scores in astragalus group,low and medium dose volatile oil from artemisia dracunculus groups were significantly lower than those of the model group (infiltration score:2.92 ± 0.39,2.95 ± 0.35,2.95 ± 0.37 vs.3.25 ± 0.45,necrosis score:2.46 ± 0.50,2.50 ± 0.51,2.54 ± 0.50 vs.2.91 ± 0.51,all P ＜0.05).Conclusions Volatile oil from artemisia dracunculus can protect cardiomyocytes by removing the virus and regulating the immune function in the body.But the protective effects of volatile oil from artemisia dracunculus is related to the dosage,and the effects of low and medium dose are better.

ObjectiveTo explore the prevalence and clinical features of hyperuricemia and its influencing factors in elderly male people aged 90 years and above. MethodsOne hundred elderly male people aged 90 years and above who underwent routine health examination in our hospital in 2007 were selected in the study. Serum uric acid level was examined by uricase-peroxidase method, and all patients were divided into hyperuricemia group and control group according to the serum uric acid level. Clinical and biochemical indications were compared between the two groups, and logistic regression was used to analyze the influencing factors of hyperuricemia in elderly people. ResultsThe serum uric acid level was increased in 20% of the elderly people, and the prevalence of gouty arthritis was 1%. The levels of blood urea nitrogen and serum creatinine were higher in hyperuricemia group than in control group[(10. 98±4.29) mmol/L vs. (6. 87± 1.86) mmol/L, (125.2±25.9)μmol/L vs. (93. 4±19. 8)μmol/L;both P<0.05)3. The patients in hyperuricemia group had a higher prevalence of hypertension and hypertriglyceridemia, and a higher proportion of diuretic application than patients in control group(P<0. 05). Logistic regression analysis showed that serum uric acid level had the most remarkable correlation with serum creatinine(OR= 1. 969), followed by fasting blood glucose (OR= 1. 310)and blood urea nitrogen(OR = 1.161). There was negative correlation between serum uric acid level and plasma cholesterol level(OR = 0. 802). ConclusionsThe prevalence of hyperuricemia is high in elderly people aged 90 years and above, while the incidence of gouty arthritis is low. Renal function impairment, metabolic syndrome and thiazide diuretic are the major factors for hyperuricemia.

Intravenous and intratracheal implantation of mesenchymal stem cells (MSCs) may offer ameliorating effects on pulmonary hypertension (PH) induced by monocrotaline (MCT) in rats. The aim of this study was to examine the anti-remodeling effect of intravenous MSCs (VMSCs) and intratracheal MSCs (TMSCs) in rats with PH, and the underlying mechanisms. MSCs were isolated from rat bone marrow and cultured. PH was induced in rats by intraperitoneal injection of MCT. One week after MCT administration, the rats were divided into 3 groups in terms of different treatments: VMSCs group (intravenous injection of MSCs), TMSCs group (intratracheal injection of MSCs), PH group (no treatment given). Those receiving saline instead of MCT served as negative control (control group). Pulmonary arterial structure was pathologically observed, pulmonary arterial dynamics measured, and remodeling-associated cytokines Smad2 and Smad3 detected in the lungs, three weeks after MCT injection. The results showed that PH group versus control group had higher pulmonary arterial pressure (PAP) and wall thickness index (WTI) 21 days after MCT treatment. The expression of phosphorylated (p)-Smad2 and the ratio of p-Smad2/Smad2 were much higher in PH group than in control group. Fluorescence-labeled MSCs were extensively distributed in rats' lungs in VMSCs and TMSCs groups 3 and 14 days after transplantation, but not found in the media of the pulmonary artery. WTI and PAP were significantly lower in both VMSCs and TMSCs groups than in PH group three weeks after MCT injection. The p-Smad2 expression and the ratio of p-Smad2/Smad2 were obviously reduced in VMSCs and TMSCs groups as compared with those in PH group. In conclusion, both intravenous and intratracheal transplantation of MSCs can attenuate PAP and pulmonary artery remodeling in MCT-induced PH rats, which may be associated with the early suppression of Smad2 phosphorylation via paracrine pathways.
Animals ; Atrial Remodeling ; drug effects ; physiology ; Hypertension, Pulmonary ; chemically induced ; physiopathology ; Male ; Mesenchymal Stromal Cells ; pathology ; Monocrotaline ; pharmacology ; Pulmonary Artery ; drug effects ; physiopathology ; Rats ; Rats, Sprague-Dawley