Triterpene esters of cirsium setosum and their anti-inflammatory activity.

Two new triterpene fatty acid esters, 3ß-palmityloxy-12,27-cyclofriedoolean-14-en-11α-ol (1) and 3ß-palmityloxy-19α-hydroxyursane (2), together with 3ß-hydroxy-11-oxo-olean-12-enyl palmitate (3) were isolated from the potent anti-inflammatory active fraction of the petroleum ether-soluble part of Cirsium setosum ethanol extract. Compound 1 was found to be a rare 12,27-cyclopropane triterpenoid. Their structures were determined through spectral data analysis combined with literature reports. Furthermore, in vitro experiment, compounds 1-3 exhibited significant inhibitory effects on nitric oxide production in lipopolysaccharide-activated mouse RAW264.7 macrophages.

Seco-nortriterpenoids from Cirsium setosum and their anti-inflammatory activity.

Five unusual seco-nortriterpenoids, 3ß-hydroxy-20,21-seco-30-nortaraxastan-20,21-dioic acid (1), 3ß-hydroxy-20,21-seco-30-nortaraxastan-20-oic-21-oate (2), 3ß-hydroxy-20-oxo-21,22-seco-30-nortaraxastan-22-oic acid (3), 3ß-hydroxy-19-oxo-20,21-seco-29,30-nortaraxastan-21-oic acid (4) and 3ß-hydroxy-19-oxo-20,21-seco-19-norlupan-21-oic acid (5) were isolated and elucidated from the anti-inflammatory activity fraction of the ethanol extract of Cirsium setosum. The structures of these compounds were established through spectroscopic methods. Preliminary biological assays showed that compounds 1-5 had significant inhibitory effect on NO production on lipopolysaccharide-stimulated RAW 264.7 cells, and compound 1 showed the strongest anti-inflammatory activity. This type of ring-opening compound is the first seco-triterpenoid structure discovered from the genus of Cirsium.

Refinement of the equine influenza model in the natural host: A meta-analysis to determine the benefits of individual nebulisation for experimental infection and vaccine evaluation in the face of decreased strain pathogenicity.

Equine Influenza (EI) is an important respiratory disease of horses caused by H3N8 equine influenza viruses (EIV). Vaccination is a key strategy to prevent or control this disease. However, EIV undergoes continuous antigenic drift and whilst numerous EI vaccines are commercially available worldwide, an accurate evaluation of their efficacy is frequently required through clinical trials conducted in the natural host. Room nebulisation is one of the chosen methods to challenge horses during EI vaccine studies. A potential decreased pathogenicity observed with recent Florida Clade 2 (FC2) EIV isolates have increased the heterogeneity of the clinical response and virus shedding measured after infection by room nebulisation, which reduced the statistical power of studies. Our objectives were to compare clinical and virological parameters following experimental infection with several different EIV strains and to confirm that individual nebulisation is a model refinement that prevents an increase of the number of animals per group. This study is a retrospective comparison and meta-analysis of clinical and virological results collected from 9 independent EIV infection studies in the natural host. Naïve Welsh mountain ponies were experimentally infected by room or individual nebulisation with FC2 EIV strains, including A/equine/Richmond/1/07 (R/07), A/equine/East Renfrewshire/11 (ER/11), A/equine/Cambremer/1/2012 (C/12) and A/equine/Northamptonshire/1/13 (N/1/13). The retrospective meta-analysis confirmed a decreased pathogenicity of the EIV ER/11 and C/12 strains when compared with R/07. Experimental infection by individual nebulisation improved the clinical and virological parameters induced by recent FC2 strains, when compared with conventional room nebulisation. In conclusion, individual nebulisation offers a better control of the challenge dose administered and a greater homogeneity of the response measured in control animals. This in turn, helps maintain the number of animals per group to the minimum necessary required to obtain meaningful results in vaccine efficacy studies, which adheres to the 3Rs (Replacement, Reduction and Refinement) principles.

[Long-term outcome of patients discharged with fever of unknown origin in the Department of General Internal Medicine of Peking Union Medical College Hospital].

OBJECTIVE: To investigate the clinical characteristics and outcome of patients with undiagnosed fever of unknown origin (FUO). METHODS: To retrospectively review the clinic data of patients discharged with FUO from the Department of General Internal Medicine, Peking Union Medical College Hospital during 2004 to 2008. Medical records and phone call follow-up data were collected until 2014. RESULTS: Among 758 in-patients diagnosed with FUO, 70 patients still discharged with FUO were enrolled in this study, including 23 males and 47 females. There were 14 missing patients. Finally, definite diagnoses were made in 20 patients by clinical reassessments, empirical therapy or repeated biopsies, in whom 3 patients dying from underlying diseases. A total of 36 patients did not get final diagnoses, while fever was relieved in 23 patients, including 10 treated with corticosteroids or non-steroid anti-inflammatory drugs (NSAIDs) from 1 month to 12 months due to suspected connective tissue diseases. Another 3 patients still had episodic fever. Seven patients died shortly after discharge. There were 3 dying in the long-term follow-up. The overall FUO-related mortality was 18.6%. Mortality was correlated with the number of dysfunctional organs, especially cytopenia, coagulation dysfunction, bleeding events, respiratory damage and acute renal failure with OR 2.1, 9.9, 3.3 and 6.6 (P < 0.05) respectively. CONCLUSIONS: Close follow-up, intermittent clinical reassessments, repeated biopsies will contribute to the diagnosis of patients discharged with FUO. Empirical therapy with corticosteroids, NSAIDs or anti-tubercular drugs in selected patients may be safe and effective. Mortality rates increased with impaired organs, especially the hematological, respiratory and renal systems.

Prognostic role of C-reactive protein and interleukin-6 in dialysis patients: a systematic review and meta-analysis.

Inflammation may be associated with mortality in dialysis patients. This study aims to summarize the prognostic value of two common inflammatory markers, C-reactive protein (CRP) and interleukin-6 (IL-6) for dialysis outcome. A total of 109 CRP studies and 22 IL-6 studies were identified from PubMed and EMBASE after systematic searching and assessment. The combined hazard ratios (HRs) of CRP and IL-6 for mortality were analyzed. For all-cause mortality (ACM), both CRP and IL-6 could significantly predict the outcome, with the pooled HRs of 1.142 (95%CI: 1.118-1.166) and 1.152 (95%CI: 1.094-1.214) for CRP and IL-6, respectively. For cardiovascular disease mortality (CVDM), the pooled HR of CRP (1.182, 95%CI: 1.134-1.232) was close to that of IL-6 (1.181, 95%CI: 1.068-1.307). Therefore, elevated levels of CRP or IL-6 were significantly associated with higher ACM and higher CVDM in dialysis patients. The predictive value of the inflammatory biomarkers may be useful in clinical practice.

Chimeric viruses containing the N-terminal ectodomains of GP5 and M proteins of porcine reproductive and respiratory syndrome virus do not change the cellular tropism of equine arteritis virus.

Equine arteritis virus (EAV) and porcine reproductive and respiratory syndrome virus (PRRSV) are members of family Arteriviridae; they are highly species specific and differ significantly in cellular tropism in cultured cells. In this study we examined the role of the two major envelope proteins (GP5 and M) of EAV and PRRSV in determining their cellular tropism. We generated three viable EAV/PRRSV chimeric viruses by swapping the N-terminal ectodomains of these two proteins from PRRSV IA1107 strain into an infectious cDNA clone of EAV (rMLVB4/5 GP5ecto, rMLVB4/5/6 Mecto and rMLVB4/5/6 GP5&Mecto). The three chimeric viruses could only infect EAV susceptible cell lines but not PRRSV susceptible cells in culture. Therefore, these data unequivocally demonstrate that the ectodomains of GP5 and M are not the major determinants of cellular tropism, further supporting the recent findings that the minor envelope proteins are the critical proteins in mediating cellular tropism (Tian et al., 2012).

Development and characterization of an infectious cDNA clone of the modified live virus vaccine strain of equine arteritis virus.

A stable full-length cDNA clone of the modified live virus (MLV) vaccine strain of equine arteritis virus (EAV) was developed. RNA transcripts generated from this plasmid (pEAVrMLV) were infectious upon transfection into mammalian cells, and the resultant recombinant virus (rMLV) had 100% nucleotide identity to the parental MLV vaccine strain of EAV. A single silent nucleotide substitution was introduced into the nucleocapsid gene (pEAVrMLVB), enabling the cloned vaccine virus (rMLVB) to be distinguished from parental MLV vaccine as well as other field and laboratory strains of EAV by using an allelic discrimination real-time reverse transcription (RT)-PCR assay. In vitro studies revealed that the cloned vaccine virus rMLVB and the parental MLV vaccine virus had identical growth kinetics and plaque morphologies in equine endothelial cells. In vivo studies confirmed that the cloned vaccine virus was very safe and induced high titers of neutralizing antibodies against EAV in experimentally immunized horses. When challenged with the heterologous EAV KY84 strain, the rMLVB vaccine virus protected immunized horses in regard to reducing the magnitude and duration of viremia and virus shedding but did not suppress the development of signs of EVA, although these were reduced in clinical severity. The vaccine clone pEAVrMLVB could be further manipulated to improve the vaccine efficacy as well as to develop a marker vaccine for serological differentiation of EAV naturally infected from vaccinated animals.

[An analysis of disease spectrum of patients admitted to the General Internal Medicine Unit at Peking Union Medical College Hospital from 2004 to 2008, and the value of general internal medicine unit in comprehensive hospitals].

OBJECTIVE: To analyze the disease spectrum of patients admitted to the General Internal Medicine Unit at Peking Union Medical College Hospital, which is the first academic division of general internal medicine in the department of medicine within Chinese medical colleges and universities, and the value of general internal medicine unit in comprehensive hospitals. METHODS: A retrospective data review of patients admitted to the General Internal Medicine Unit from 2004 to 2008 was conducted from hospital information system and partially by chart review manually. Analysis of disease spectrum was performed thereafter. RESULTS: A total of 2593 patients were included in our study. It consisted of 1075 men and 1518 women, with an average age of 45.1 years old. Forty point three percent of these patients were from Beijing, the local city, and the remaining 59.7% were from outside of Beijing. Sixty-four point nine percent (1683/2593) of these patients did not have a clear diagnosis on admission, including 758 fever of unknown origin (FUO) cases and 925 non-FUO cases. The final diagnostic rate of the FUO cases was 89.2% [676/758, with the first three leading causes as diseases of the musculoskeletal system and connective tissue (29.8%), certain infectious and parasitic diseases (26.3%), and neoplasm (14.5%)]. The final diagnostic rate of the 928 non-FUO cases was 86.8% (803/925), with the first three leading causes as musculoskeletal system and connective tissue (24.9%), neoplasm (15.5%), and diseases of blood and blood-forming organs (11.4%). Despite most diagnoses fitting into the above categories, the array of diseases was broad with as many as 550 discharge diagnoses from 2004 to 2008. CONCLUSIONS: During 2004 - 2008, there was a high proportion of cases that presented to the General Internal Medicine Unit at Peking Union Medical College Hospital with an unclear diagnosis, and the spectrum of diseases diagnosed was very broad. This kind of patient admitting model might not only benefit patients with no clear admission diagnosis and patients with multidisciplinary medical problems for whom it is usually difficult to be admitted by a specialty unit, but would also benefit medical students and residents by providing a good clinical medicine teaching base. These features show the value of general internal unit in comprehensive hospitals.

Prognostic role of systemic inflammatory response in renal cell carcinoma: a systematic review and meta-analysis.

PURPOSE: To summarize the global predicting role of systemic inflammatory response for survival in renal cell carcinoma. METHODS: Eligible studies were identified and assessed for quality through multiple search strategies. Data were collected from studies comparing overall, cancer-specific or relapse-free survival in patients with elevated C-reactive protein (CRP), platelet count (PC) and erythrocyte sedimentation rate (ESR) levels with those with lower levels. Studies were pooled, and combined hazard ratios (HRs) of CRP, PC and ESR for survival were calculated, respectively. RESULTS: A total of 47 studies were included for meta-analysis (18 for CRP, 15 for PC and 14 for ESR). For overall survival, the pooled HR of CRP (3.23, 95% CI: 1.77, 5.89) was higher than that of PC (1.96, 95% CI: 1.40, 2.75) and that of ESR (1.86, 95% CI: 1.34, 2.59). These indicators were strong predictors for cancer-specific survival with individual HRs being 3.46 (95% CI, 2.80, 4.27) of CRP, 3.22 (95% CI, 2.25, 4.62) of PC, and 3.85 (95% CI, 3.31, 4.48) of ESR, respectively. All three inflammatory indictors also predicted relapse-free survival (HRs > 2.0). CONCLUSION: The systemic inflammatory response predicted poor survival in patients with renal cell carcinoma.

[Cardiac valve involvement in Behcet's disease: a clinical study of 10 patients].

OBJECTIVE: To assess the clinical features in patients with cardiac valve lesions associated with Behcet's disease (BD). METHODS: We retrospectively reviewed the clinical data of 10 BD patients with cardiac valve lesions who were admitted to Peking Union Medical College Hospital (PUMCH) during June 1999 to June 2009. RESULTS: Aortic regurgitation occurred in 6% of patients with BD in PUCMH. Patients included 8 male and 2 female with the mean age of 36.5. All the patients had occult onset cardiac symptoms with an average length of clinical course of 6 years. 5 patients fulfilled the ISG diagnostic criteria for BD and another 5 patients diagnosed by experts. The main echocardiography findings were severe aortic regurgitation, aneurysmal dilatations of ascendant aorta, echo-free space at the aortic root, aortic valve prolapse, mesh like mass incorporating aortic cusp, aortic valve perforation, et al. 3 patients underwent 7 operations. 5 simple aortic valve replacement (AVR) surgeries resulted in severe perivalvular leakage. 2 patients underwent Bentall and heart transplant surgeries respectively with perioperative immunosuppressive therapy had no complications. CONCLUSION: Cardiac valve involvement in BD is a rare but critical problem that requires a timely diagnosis and management. The current diagnostic criteria may have possibilities of delayed diagnosis of such problem. Echocardiography seems to be helpful for the timely diagnosis. The immunosuppressive therapy and Bentall type operations may be essential for improving the treatment outcome of BD with cardiac valve lesions.

The impacts of thiazolidinediones on circulating C-reactive protein levels in different diseases: a meta-analysis.

OBJECTIVES: Thiazolidinediones (TZDs), including rosiglitazone and pioglitazone, are widely used as antidiabetic agents. Here we present an updated meta-analysis of the respective effects of rosiglitazone and pioglitazone on the levels of C-reactive protein (CRP), a biomarker and predictor of CAD risks. METHODS AND RESULTS: PubMed and EMBASE were systematically searched using the terms "C-reactive protein" and "rosiglitazone" or "pioglitazone" or "thiazolidinedione". A total of 59 rosiglitazone studies and 36 pioglitazone studies were included for evaluation. These were classified as 2-group studies and 1-group studies according to their study design. Standard mean difference (SMD) of CRP and 95% CI were calculated to evaluate the effect of TZDs on CRP. The overall SMD for rosiglitazone treatment is -0.392 (95% CI, [-0.446, -0.338]) in 2-group studies and -0.424 (95% CI, [-0.501, -0.346]) in 1-group studies. The overall SMD for pioglitazone treatment is -0.577 (95% CI, [-0.732, -0.421]) in 2-group studies and -0.327 (95% CI, [-0.439, -0.216]) in 1-group studies. Moreover, TZDs were found to significantly reduce CRP levels in both diabetes mellitus (DM) and non-DM patients. CONCLUSIONS: Our meta-analysis suggested that both rosiglitazone and pioglitazone significantly decrease serum CRP levels. TZDs presented their CRP-lowering effect in both DM and non-DM patients.

Association between serum amyloid A and obesity: a meta-analysis and systematic review.

BACKGROUND: Emerging evidence indicates an association of the acute-phase protein serum amyloid A (SAA) with obesity. Here we review and summarize quantitatively the available data related to this association. METHODS: PubMed was systematically searched using the terms "serum amyloid A" and "obesity." Eighty-one relevant studies between January 1966 and July 2009 were identified. Of these, only 11 cross-sectional studies and 10 prospective studies with successful interventions met our inclusion criteria for the meta-analysis. All analyses were conducted using the Comprehensive Meta-Analysis software. Literature pertaining to the relationship between SAA and other inflammatory markers, and the association between SAA and obesity-related disorders, such as cardiovascular diseases, atherosclerosis, diabetes, and insulin resistance was also reviewed. RESULTS: A strong association between body mass index and SAA levels was found in the 11 cross-sectional studies. The overall correlation coefficient is 0.230 (95% CI 0.160-0.297, P < 0.0005). The ten prospective studies were subsequently analyzed, and the difference in SAA levels before and after weight loss, expressed as standardized mean difference was -0.480 (95% CI -0.678 to -0.283, P < 0.0005). We discuss some potential underlying mechanisms and clinical applications for reducing SAA levels in obesity.

Dual-subtype feline immunodeficiency virus vaccine provides 12 months of protective immunity against heterologous challenge.

The duration of immunity of the dual-subtype feline immunodeficiency virus (FIV) vaccine, Fel-O-Vax FIV, for protection against subtype-B FIV was assessed in this study. Vaccinated cats along with controls were challenged with FIV(FC1), a subtype-B FIV strain, 54 weeks after the final vaccination, and monitored for 46-48 weeks for provirus and viral RNA in peripheral blood, provirus in lymphoid organs, and CD4:CD8 ratios. Results of provirus detection in peripheral blood and lymphoid organs and plasma viral RNA loads showed that 10/14 vaccinated cats were fully protected for 48 weeks against infection with FIV(FC1) whereas 5/5 controls were persistently infected with FIV(FC1). CD4:CD8 inversions were noted in association with FIV infection and viral loads were not significantly different between FIV infected controls and the unprotected vaccinated animals.

A dual-strain feline calicivirus vaccine stimulates broader cross-neutralization antibodies than a single-strain vaccine and lessens clinical signs in vaccinated cats when challenged with a homologous feline calicivirus strain associated with virulent systemic disease.

Feline calicivirus (FCV) causes an array of clinical disease in cats. Traditionally this disease has been associated with respiratory disease, limping, or chronic stomatitis. Within the last 10 years, virulent systemic feline calicivirus (VS-FCV) has been recognized which causes additional clinical signs and has a higher fatality rate. A dual-strain FCV vaccine containing a strain of FCV associated with traditional respiratory disease and a VS-FCV strain stimulates serum cross-neutralization antibodies when tested against field strains from Europe and VS-FCV strains from USA. Following challenge with a homologous VS-FCV strain, vaccinated cats had significantly reduced clinical signs.

Serological differentiation of FIV-infected cats from dual-subtype feline immunodeficiency virus vaccine (Fel-O-Vax FIV) inoculated cats.

Feline immunodeficiency virus (FIV) vaccine, Fel-O-Vax FIV, was released for sale in the US in 2002. The antibodies of vaccinated cats interfere with serological assays by currently available FIV diagnostic kits. In this study, we investigated whether it is possible to distinguish serologically cats vaccinated with Fel-O-Vax FIV from cats experimentally or naturally infected with FIV. A total of 153 sera taken from 97 cats were used as serum samples. Enzyme linked immunosorbent assay (ELISA) was performed using whole FIV antigen and formalin treated whole FIV antigen, recombinant-gag (r-gag) antigen, and transmembrane (TM) peptide. Statistical analysis was performed using ELISA optical density (O.D.) values obtained with each antigen as variables. Except for the ELISA O.D. values obtained with r-gag antigen, a significant difference in ELISA O.D. values was observed between the vaccinated and the infected groups. However, it was not possible to distinguish both groups unequivocally. Using discriminant analysis, it was possible to distinguish the two groups with an accuracy of 97.1% with two discriminating variables (ELISA O.D. values obtained with formalin treated whole FIV antigen, and TM peptide), 97.8% with three discriminating variables (ELISA O.D. values obtained with whole FIV antigen, formalin treated whole FIV antigen, and TM peptide). Therefore, it was considered possible to distinguish cats vaccinated with Fel-O-Vax FIV from FIV-infected cats by ELISA using two types of antigens including formalin treated whole FIV antigen and TM peptide, or three types of antigens including formalin treated whole FIV antigen, TM peptide and whole FIV antigen.

Dual-subtype vaccine (Fel-O-Vax FIV) protects cats against contact challenge with heterologous subtype B FIV infected cats.

Fel-O-Vax FIV is a dual-subtype vaccine consisting of inactivated whole viruses of subtype A (Petaluma strain) and subtype D (Shizuoka strain). The efficacy of this vaccine against heterologous subtype A strain challenge was demonstrated, but it is unclear whether the result reflects efficacy in the field. In this study, we evaluated the efficacy of this vaccine against contact challenge by exposing both vaccinated and unvaccinated control animals with cats infected with Aomori-2 strain belonging to subtype B, a subtype prevalent in many regions of the world. Nineteen specific-pathogen-free (SPF) cats were divided into a vaccinated group (six cats), an unvaccinated control group (eight cats), and a challenge group (five cats), and maintained in the same room. Cats were monitored for FIV proviral DNA by nested PCR and for FIV-specific antibody levels by ELISA. After 1 year of commingling, each cat in the vaccinated group was given a booster dose. In addition, the original challenge group was removed and replaced with another challenge group of SPF cats, which were inoculated with the Aomori-2 strain. FIV infection was confirmed in four of the eight animals in the unvaccinated control group by the 29th week in the second year of commingling. In contrast, all of the animals were negative in the vaccinated group. These findings confirmed the efficacy of this vaccine against heterologous stains classified as subtype B, and suggested that the vaccine exhibits broad efficacy against genetically diverse FIV.

Efficacy and safety of a feline immunodeficiency virus vaccine.

Fel-O-Vax FIV is an inactivated virus vaccine designed as an aid in the prevention of infection of cats, 8 weeks or older, by feline immunodeficiency virus (FIV). It contains two genetically distinct FIV strains. The efficacy of this vaccine was demonstrated in a vaccination-challenge study designed to meet various regulatory requirements for registering the vaccine. Eight-week-old kittens were vaccinated with an immunogenicity vaccine which contained minimal release levels of FIV antigens formulated with a proprietary adjuvant system. Twelve months later, all vaccinates and controls were challenged with a heterologous FIV strain. Following the vigorous challenge exposure, cats were monitored for FIV viremia. It was found that 16% of the vaccinated cats developed viremia while 90% of the controls became persistently infected with FIV, which demonstrated that the vaccine was efficacious and the protective immunity lasted for at least 12 months. The safety of the vaccine was demonstrated by a field safety trial in which only 22 mild reactions of short duration were observed following administering 2051 doses of two pre-licensing serials of Fel-O-Vax FIV to cats of various breeds, ages and vaccination histories. Thus, Fel-O-Vax FIV is safe and efficacious for the prevention of FIV infection in cats.