Peripheral Blood Leukocyte Count Research Articles

Usefulness of Various Peripheral Blood Leukocyte Count ratios in Malaria Evaluation

Malaria is a major public health problem in India. Changes in blood cell counts are a wellknown feature of malaria. Hence an attempt has been made to assess the role of leucocyte ratios in malaria as predictors of malaria infection and its clinical severity. Methods: A retrospective observational cohort study was carried out in a tertiary care teaching hospital over a period of 2 months. A total of 171 samples and 48 controls were included in the study. All the absolute and differential white blood cell counts were done. The neutrophillymphocyte count ratio (NLCR), monocyte-lymphocyte count ratio (MLCR) and monocyteneutrophil count ratio (MNCR) were calculated by using the absolute neutrophil, lymphocyte and monocyte count respectively. The results were analyzed statistically using SPSS software for windows. Results: Among 171 cases 145(84.8%), 19(11.1%), 7(4.1%) were Pl. vivax, Pl. falciparum and mixed malaria respectively. 52 cases were severe and 119 were non-severe. 52, 45, 47, 27 were parasitemia grade 1, 2, 3, 4 respectively. NLCR, MLCR and MNCR showed a sensitivity of 60.8%, 71.7%, 70.8% and specificity of 43.8%, 60.4%, and 56.3% respectively in malaria diagnosis. There was no association of NLCR, MLCR, MNCR with the type and severity of malaria. NLCR, MNCR correlated with varying grade of parasitemia. Conclusion: The results show the ratios NLCR, MLCR and MNCR have a better sensitivity but less specificity. The sensitivity and specificity for MLCR are the highest and can be used as a supportive to a screening test. The ratios can be used to show the presence of malaria parasite but there is no association with the type and severity of malaria. MLCR is a good predictor of malarial presence. NLCR and MNCR are associated with parasitemia.

Usefulness of Various Peripheral Blood Leukocyte Count ratios in Malaria Evaluation

Usefulness of Various Peripheral Blood Leukocyte Count ratios in Malaria Evaluation

Correlation of SOCS3 mRNA Expression Level with Clinical Characteristics and Early Response to Treatment in Pediatric Acute Lymphoblastic Leukemia

The expression levels of SOCS3 mRNA in bone marrow mononuclear cells from 45 cases of newly diagnosed ALL at initial diganosis and induction remission and 13 normal children as controls were detected by SYBR Green fluorescence quantitative PCR; the correlation of SOCS3 mRNA expression level with risk and clinical characteristics of ALL was analyzed by means of statistical method; the immunofluorescence histochemistry method and laser confocal microscopy were used to detect the sites and expression level of SOCS3 mRNA in bone marrow samples of ALL patients. The SOCS3 mRNA expression level at initial diagnosis of 45 ALL patients was significantly lower than that of normal controls (P<0.05), and that in induction remission of 45 patients was not significant different from normal controls (P<0.05); the SOCS3 mRNA expression level at initial diagnosis of patients with standasd and high risk was higher than that of patients with low risk (P<0.05). The 45 patients were divided into high expression and low expression groups according to SOCS3 mRNA expression level at initial diagnosis by median method, comparison of clinical characteristics in 2 groups was found that the SOCS3 mRNA high expression group had even more higher leukocyte count in peripheral blood, even more higher LDH level and much more poor prognostic genes; in addition, the high expression group showed more higher risk in comparison between 2 group. The SOCS3 mRNA expression is down-regulated at initial diagnosis, while recovered to normal level after induction remission of disease, thus the SOCS3 can be used as an indicator for evaluation of disease status. The high expression of SOCS3 mRNA up-regulates the disease risk, therefore the SOCS3 mRNA can be used as a factor for evaluation of ALL risk. The treatment of ALL via regulation of SOCS3 mRNA expression level maybe can become a new way. The regulation of SOCS3 mRNA expression level maybe can become a new way for treatment of ALL.

Rice protein prolamin promotes anti-leukemia immunity and inhibits leukemia growth in vivo

Prolamin is a heat-stable storage protein of rice (Oryza sativa). This study aimed to examine the effect of prolamin on anti-tumor immune response in vitro and leukemia growth in vivo. The prolamin-enriched rice fractions were prepared to stimulate peripheral blood mononuclear cells (MNC) from mice spleen. The MNC-conditioned medium (MNC-CM) was collected to treat leukemia L1210 cells. Human MNC-CM was prepared to treat Jurkat acute T cell leukemia cells. Purified prolamin was orally administered to syngeneic L1210-bearing DBA/2 mice to assess weights of tumor, liver and spleen, liver histopathology, peripheral blood neutrophil count and cytokine levels. Prolamin-prepared MNC-CM inhibited the viability of murine leukemia L1210 cells and human leukemia Jurkat cells, indicating an immunomodulatory effect. In syngeneic L1210-bearing DBA/2 mice, oral administration of purified prolamin dose-dependently decreased the tumor weight and attenuated the leukemia-induced reduction of liver and spleen weights. Prolamin inhibited the increase of peripheral blood leukocyte count. The levels of tumor necrosis factor-α and interferon-γ in MNC-CM and mice serum were significantly increased by prolamin treatment. No significant change in body weight, serum alanine aminotransferase and creatinine levels was noted by prolamin treatment. Rice prolamin could effectively promote anti-tumor immunity and inhibit leukemia growth without significant toxicity.

Photodynamic therapy controls of Staphylococcus aureus intradermal infection in mice.

Infections caused by Staphylococcus aureus lead to skin infections, as well as soft tissues and bone infections. Given the communal resistance to antibiotics developed by strains of this bacterium, photodynamic therapy emerges as a promising alternative treatment to control and cure infections. Females of the Balb/C mice were infected with 108CFU of methicillin-resistant S. aureus (MRSA) and divided into four distinct groups: P-L- (negative control group), P+L- (group exposed only to curcumin), P-L+ (group exposed only to LED incidence of 450nm, 75mW/cm2, and 54J/cm2 for 10min), and P+L+ (group exposed to curcumin followed by 10min of LED irradiation) (n=24). The mice were euthanized 48 and 72h after infection, and biologic materials were collected for analysis of the bacterial load, peripheral blood leukocyte counts, and draining lymph nodes cell counts. The normalization of data was checked and the ANOVA test was applied. The bacterial load in the draining lymph node of P+L+ group was lower when compared to the control groups 72h post infection (p<0.0001), indicating that the LED incidence associated with curcumin controls of the staphylococci intradermal infection. The number of the total lymph node cells shows to be lower than control groups in the two availed times (p<0.01). The histological analysis and the counting of white blood cells did not show differences among cells in the blood and in the tissue of infection. This is the first report showing that photodynamic therapy may be effective against MRSA infection in a murine model of intradermal infection.

Chronic Pseudomonas aeruginosa biofilm infection impairs murine S100A8/A9 and neutrophil effector cytokines-implications for delayed wound closure?

The impact of Pseudomonas aeruginosa biofilm infections in chronic wounds and clinical implication for healing is receiving increased attention. However, the pathophysiology of host/pathogen interplay is not fully understood. By further revealing the mechanisms, necessary new treatment strategies may be identified. Since the background for chronic wounds is diverse, representative animal models are important. We assessed host response and spontaneous wound closure in the relatively resistant C3H/HeN and the susceptible BALB/c mouse strain. Full-thickness burn wounds were inflicted in 108 mice. Pseudomonas aeruginosa biofilm (106 colony forming units) was injected subcutaneously in 72 mice, euthanised day 4, 7 or 10 days post-infection. Wounds were analysed for neutrophil host response markers: S100A8/A9, keratinocyte-derived chemokine and granulocyte-colony stimulating factor. Total peripheral blood leucocyte and polymorphonuclear count were assessed in parallel. Histopathology evaluated wound inflammatory burden. Photoplanimetry described macroscopical wound closure. Stable chronic wound infection was established in all challenged mice. Pseudomonas aeruginosa biofilm suppressed neutrophil host response in wounds. C3H/HeN mice achieved earlier systemic inflammatory control and healed faster than BALB/c mice. Pseudomonas aeruginosa biofilms perturb host defence thereby inducing a steady state of chronic infection which may impair wound healing. These results indicate therapeutic options for immune modulation of biofilm-infected wounds.

Serum proteins and lipids in mild form of calf bronchopneumonia: candidates for reliable biomarkers

AbstractCalf bronchopneumonia is complex multifactorial disease and for its accurate diagnosis and therapy, besides clinical examination, microbiologic, hematologic and biochemical analyses could be necessary. In general, additional analyses are not implemented, mainly because the disease biomarkers are not defined.To establish which analysis might be useful for determining the severity of the disease, we analyzed 23 three-month old calves with mild clinical signs of bronchopneumonia and 15 age-matched healthy calves.Pasteurella multocidawas isolated from deep nasal swabs of diseased calves. Peripheral blood erythrocyte and leukocyte count of bronchopneumonic and healthy calves showed no difference. Serum proteins, lipoproteins and lipids were analyzed with spectrophotometry, agarose gel electrophoresis, non-reducing SDS-PAGE, gel zymography, and thin-layer chromatography. The bronchopneumonic calves had an increased level of circulating immune complexes and α globulins, which contain some of the positive acute phase proteins. In diseased calves the increased concentration of total γ globulins (IgG), due to an increased concentration of anionic γ globulins (predominately IgG1), was detected. The increased concentration of anionic γ globulins followed by increased concentration of transferrin (negative acute phase protein) and HDL cholesterol, decreased concentration of LDL-cholesterol, unchanged activity of matrix metalloproteases and leukocyte counts might reflect the obvious absence of generalized inflammation. A positive correlation was found between the acquired results and the appearance of mild clinical signs. Therefore, we believe that the parameters analyzed in the peripheral blood could be applied as reliable disease markers to distinguish between severe (inflammatory) and mild forms of calf bronchopneumonia and to predict a better outcome for these calves.

Management of Clostridium difficile Infection in Inflammatory Bowel Disease: Expert Review from the Clinical Practice Updates Committee of the AGA Institute.

The purpose of this expert review is to synthesize the existing evidence on the management of Clostridium difficile infection in patients with underlying inflammatory bowel disease. The evidence reviewed in this article is a summation of relevant scientific publications, expert opinion statements, and current practice guidelines. This review is a summary of expert opinion in the field without a formal systematic review of evidence. Best Practice Advice 1: Clinicians should test patients who present with a flare of underlying inflammatory bowel disease for Clostridium difficile infection. Best Practice Advice 2: Clinicians should screen for recurrent C difficile infection if diarrhea or other symptoms of colitis persist or return after antibiotic treatment for C difficile infection. Best Practice Advice 3: Clinicians should consider treating Cdifficile infection in inflammatory bowel disease patients with vancomycin instead of metronidazole. Best Practice Advice 4: Clinicians strongly should consider hospitalization for close monitoring and aggressive management for inflammatory bowel disease patients with C difficile infection who have profuse diarrhea, severe abdominal pain, a markedly increased peripheral blood leukocyte count, or other evidence of sepsis. Best Practice Advice 5: Clinicians may postpone escalation of steroids and other immunosuppression agents during acute C difficile infection until therapy for C difficile infection has been initiated. However, the decision to withhold or continue immunosuppression in inflammatory bowel disease patients with C difficile infection should be individualized because there is insufficient existing robust literature on which to develop firm recommendations. Best Practice Advice 6: Clinicians should offer a referral for fecal microbiota transplantation to inflammatory bowel disease patients with recurrent C difficile infection.

Postoperative deep shoulder infections following rotator cuff repair.

Rotator cuff repair (RCR) is one of the most commonly performed surgical procedures in orthopaedic surgery. The reported incidence of deep soft-tissue infections after RCR ranges between 0.3% and 1.9%. Deep shoulder infection after RCR appears uncommon, but the actual incidence may be higher as many cases may go unreported. Clinical presentation may include increasing shoulder pain and stiffness, high temperature, local erythema, swelling, warmth, and fibrinous exudate. Generalized fatigue and signs of sepsis may be present in severe cases. Varying clinical presentation coupled with a low index of suspicion may result in delayed diagnosis. Laboratory findings include high erythrocyte sedimentation rate and C-reactive protein level, and, rarely, abnormal peripheral blood leucocyte count. Aspiration of glenohumeral joint synovial fluid with analysis of cell count, gram staining and culture should be performed in all patients suspected with deep shoulder infection after RCR. The most commonly isolated pathogens are Propionibacterium acnes, Staphylococcus epidermidis, and Staphylococcus aureus. Management of a deep soft-tissue infection of the shoulder after RCR involves surgical debridement with lavage and long-term intravenous antibiotic treatment based on the pathogen identified. Although deep shoulder infection after RCR is usually successfully treated, complications of this condition can be devastating. Prolonged course of intravenous antibiotic treatment, extensive soft-tissue destruction and adhesions may result in substantially diminished functional outcomes.

FLT3-ITD As a Prognostic Factor for Chinese Patients with Acute Myeloid Leukemia

Internal tandem duplication of the FLT3 gene (FLT3-ITD) is one of most common genetic mutations found in patients with acute myeloid leukemia (AML). Although FLT3-ITD has been widely studied and used as an important prognostic indicator of AML in the western world, its association with disease features and prognosis in Chinese AML patients to date has rarely been investigated.In this study, 177 Chinese patients with AML were enrolled including a FLT3-ITD mutation positive group (n = 83) and a FLT3-ITD mutation negative group (n = 94).In FLT3-ITD-positive AML patients, the number of patients whose peripheral blood leucocyte count (≥ 100 × 109/L) before treatment was significantly higher than in FLT3-ITD negative patients (32.1% vs 7.6%), but comparing to FLT3-ITD mutation negative patients, 1 year and 2 year event free survival rates (EFSR) of FLT3-ITD-positive AML patients was lower ((37.0% vs 56.0%) and (14.8% vs 38.4%)), and also their 1 year and 2 year OS rates were significantly lower ( (50% vs 100%) and (18.0% vs 36.5%)). However, it was found that the rate of induced remissions in FLT3-ITD-positive AML patients was much lower than in negative patients (39.4% vs 56.0%). The median survival time of the FLT3-ITD+/NPM1- group (n = 39) was superior to the FLT3-ITD+/NPM1+ group (n = 27) (16.1 months vs 5.8 months). Lastly, after comparing chemotherapy regimes, we found that the allogeneic hematopoietic stem cell transplantation treatment method significantly improved the survival rate of FLT-ITD positive AML patients (not reached vs 14.5 months). Therefore, for FLT3-ITD-positive Chinese patients with AML, the relative rate of complication with leucocyte was increased; both the event free and overall survival rates were lower, these characteristics are the same as the FLT3-ITD-positive patients in western countries. The lower induced remission rate and shorter survival time suggested FLT3-ITD+/NPM1+ mutation is inferior to FLT3-ITD+/NPM1- mutation, which is not consistant with most reports. In addition, allogeneic hematopoietic stem cell transplantation was clearly an effective treatment for patients with FLT-ITD positive AML in China. DisclosuresNo relevant conflicts of interest to declare.

Inverse PAR1 Activity of Hematopoietic Stem Cells and BM Stromal Cells Mediates G-CSF-Induced Mobilization By Regulation of Nitric Oxide Generation

Hematopoietic stem and progenitor cell (HSPC) egress from the bone marrow (BM) to the circulation is tightly regulated and is accelerated during stress conditions, a process utilized for BM harvest. Recently, we demonstrated that mouse long term repopulating hematopoietic stem cell (LT-HSC) BM retention and their rapid release to the blood circulation are governed by a switch in nitric oxide (NO) generation via distinct coagulation-related protease activated receptor 1 (PAR1) cascades (Gur-Cohen S. et al., NM, 2016).Herein we report that surface PAR1 expression can be exploited and serve as a positive predictive marker for the efficiency of human CD34+ HSPC mobilization among healthy donors in clinical G-CSF-induced mobilization for matched allogeneic transplantations. We found that PAR1 expression on circulating leukocytes before G-CSF administration was positively correlated with higher yields of mobilized leukocytes after stimulation (P<0.001). In addition, the percentage of PAR1-expressing CD34+ HSPCs in the blood and their absolute numbers before initiation of mobilization positively correlated with peripheral blood leukocyte counts (P<0.001) and yield of mobilized CD34+ HSPCs (P=0.002). Importantly, tracing the consequences of the follow-up BM transplantation, we found accelerated neutrophil recovery in patients transplanted with mobilized cells expressing higher surface PAR1 levels at baseline (P=0.074), shortening the time for neutrophil recovery (neutrophil count>0.5 x 109/L) from an average of 14.6 days to an average of 11 days post-transplant. In addition, a trend of accelerated platelet production was documented to be related with higher PAR1 expression by circulating leukocytes prior to G-CSF stimulation. Consequently, poor mobilizer donors were characterized by extremely low surface PAR1 expression on circulating CD34+ cells prior to G-CSF stimulations. Herein we present a case report of a thrombophilic donor carrying the MTHFR mutation, expressing exceedingly low PAR1 levels at baseline, with the outcome of inadequate numbers of mobilized CD34+ HSPC in the blood following G-CSF treatments.To further gain insight into the role played by PAR1 signaling in the regulation of G-CSF-induced HSPC mobilization, we used mice as a functional preclinical small animal model. We found that antagonizing PAR1 signaling attenuated both steady state release and G-CSF-induced HSPC mobilization. Furthermore, co-administration of G-CSF with PAR1 antagonist attenuated secretion of BM stromal CXCL12 and abrogated upregulation of surface CXCR4 and PAR1 expression by BM HSPCs, all leading to significantly reduced HSPC migration, differentiation and mobilization. In support, PAR1-/- mice failed to efficiently mobilize HSPCs in response to G-CSF compared to wild type counterparts. Enforced HSPC recruitment by G-CSF treatments dramatically accelerated PAR1-dependent NO production by eNOS, known to promote TACE-mediated EPCR shedding and rapid LT-HSC mobilization. Concomitantly, circulating steady state and G-CSF-mobilized stem cells lack surface EPCR expression. Intriguingly, while EPCR expression by primitive BM stem cells was transiently reduced after G-CSF treatments, antagonizing PAR1 signaling along with G-CSF stimuli blocked NO generation and synchronically expanded BM EPCR+ LT-HSC and their supportive stromal progenitor cells (MSPCs), as confirmed by increased repopulation in transplanted mice. Finally, we report an inverse PAR1 expression and regulation by BM HSPC and stromal MSPCs in mediating G-CSF-induced mobilization. G-CSF induced elevation of PAR1 expression on BM HSPCs, providing the driving force for their enhanced NO mediated migration, proliferation, differentiation and recruitment to the circulation. Contrary, the levels of PAR1 expression were reduced on MSPCs in response to G-CSF treatment, and following NO generation by eNOS activity and CXCL12 secretion, resulted in reduced mesenchymal differentiation leading to accumulated numbers of immature mesenchymal (CFU-F) and osteoblast (CFU-OB) progenitor cells.Taken together, our study identifies and highlights inverse PAR1 signaling and NO generation as essential regulator of G-CSF induced HSPC mobilization and MSPC development opening new avenues to advance therapeutics for enhancing clinical G-CSF induced stem cell mobilization and transplantation protocols. DisclosuresRuf:Iconic Therapeutics: Consultancy.

Adjusting MtDNA Quantification in Whole Blood for Peripheral Blood Platelet and Leukocyte Counts.

Alterations of mitochondrial DNA copy number (mtDNAcn) in the blood (mitochondrial to nuclear DNA ratio) appear associated with several systemic diseases, including primary mitochondrial disorders, carcinogenesis, and hematologic diseases. Measuring mtDNAcn in DNA extracted from whole blood (WB) instead of from peripheral blood mononuclear cells or buffy coat may yield different results due to mitochondrial DNA present in platelets. The aim of this work is to quantify the contribution of platelets to mtDNAcn in whole blood [mtDNAcn(WB)] and to propose a correction formula to estimate leukocytes' mtDNAcn [mtDNAcn(L)] from mtDNAcn(WB). Blood samples from 10 healthy adults were combined with platelet-enriched plasma and saline solution to produce artificial blood preparations. Aliquots of each sample were combined with five different platelet concentrations. In 46 of these blood preparations, mtDNAcn was measured by qPCR. MtDNAcn(WB) increased 1.07 (95%CI 0.86, 1.29; p<0.001) per 1000 platelets present in the preparation. We proved that leukocyte count should also be taken into account as mtDNAcn(WB) was inversely associated with leukocyte count; it increased 1.10 (95%CI 0.95, 1.25, p<0.001) per unit increase of the ratio between platelet and leukocyte counts. If hematological measurements are available, subtracting 1.10 the platelets/leukocyte ratio from mtDNAcn(WB) may serve as an estimation for mtDNAcn(L). Both platelet and leukocyte counts in the sample are important sources of variation if comparing mtDNAcn among groups of patients when mtDNAcn is measured in DNA extracted from whole blood. Not taking the platelet/leukocyte ratio into account in whole blood measurements, may lead to overestimation and misclassification if interpreted as leukocytes' mtDNAcn.

Role of Monocyte Count and Neutrophil-to-Lymphocyte Ratio in Survival of Oral Cancer Patients.

Introduction Inflammation seems to play a critical role in the development and progression of numerous cancers. Peripheral blood leukocyte count is an easily assessable parameter of systemic inflammatory response. Objective The aim of this study was to investigate whether the pretreatment leukocyte counts can predict the prognosis of patients with oral cavity cancer. Methods Medical records of 471 oral cavity cancer patients diagnosed between January 2007 and December 2008 were retrospectively analyzed. Receiver operating characteristic curve analysis and Cox proportional hazards analyses were applied to evaluate the associations of leukocyte counts with overall survival. Results The overall five year's survival of the cohort was found to be 49.4%. On univariate analysis, elevated monocyte count (≥500/mm3) and neutrophil-to-lymphocyte ratio (NLR) (>2.38) were associated with poor overall survival (OS) (p = 0.001 and 0.000, respectively). Multivariate Cox proportional hazard analysis showed that higher monocyte and NLR levels were significant independent predictors of worse OS (HR = 1.385, 95% CI = 1.049 - 1.829; p < 0.05 and HR = 1.392, 95% CI = 1.045 - 1.855; p < 0.05, respectively). The advanced overall stage and lymph nodal involvement were also independent indicators for poor OS. Conclusions Higher pretreatment monocyte and NLR levels are independent predictors of poor prognosis for patients with oral cavity cancer. Thus, these easily accessed variables can serve as a potent marker to predict the outcomes of oral cancer patients.

Predictive and Prognostic Clinical Variables in Cancer Patients Treated With Adenoviral Oncolytic Immunotherapy.

The development of oncolytic viruses has recently made great progress towards being available to cancer patients. With the breakthrough into clinics, it is crucial to analyze the existing clinical experience and use it as a basis for treatment improvements. Here, we report clinical data from 290 patients treated with oncolytic adenovirus. Using clinical variables and treatment characteristics, we constructed statistical models with regard to treatment response and overall survival (OS). Additionally, we investigated effects of neutralizing antibodies, tumor burden, and peripheral blood leucocyte counts on these outcomes. We found the absence of liver metastases to correlate with an improved rate of disease control (P = 0.021). In multivariate evaluation, patients treated with viruses coding for immunostimulatory granulocyte macrophage colony-stimulating factor were linked to better prognosis (hazard ratio (HR) 0.378, P < 0.001), as well as women with any cancer type (HR 0.694, P = 0.017). In multivariate analysis for imaging response, patients treated via intraperitoneal injection were more likely to achieve disease control (odds ratio (OR) 3.246, P = 0.027). Patients with low neutrophil-to-lymphocyte ratio before treatment had significantly longer OS (P < 0.001). These findings could explain some of the variation seen in treatment outcomes after virotherapy. Furthermore, the results offer hypotheses for treatment optimization and patient selection in oncolytic adenovirus immunotherapy.

Correlation study between complement C3 level and degree of disease activity in patients with systemic lupus erythematosus

Objective To investigate the relationship between complement C3 level and the degree of disease activity in patients with systemic lupus erythematosus(SLE). Methods A total of 1 012 patients with SLE were enrolled in this study from January 2006 to December 2013 at department of rheumatology and immunology, the first affiliated hospital of Anhui medical university. Serum complement C3 level was detected by rate erythenephelometry assay. The relationship between reduction of complement C3 and SLE was analyzed. Results Serum complement C3 clearly decreased in 782 patients, accounted for 77.27%. There were significant differences concerning serum complement C3 level among different groups of disease activity (F=131.275, P<0.01). The low serum complement C3 level was correlated with the high degree of disease activity (r=-0.517, P<0.01). Patients with SLE had a severe disease activity (SLE disease activity index ≥15) when the level of serum complement C3 was less than 0.57 g/L. Serum complement C3 levels were positively correlated with serum complement C4 (r=0.845, P<0.01), peripheral blood leukocyte count (r=0.115, P<0.01), hemoglobin (r=0.069, P<0.01), platelet count (r=0.177, P<0.01) and albumin level (r=0.091, P<0.01). There was also a negative association of serum complement C3 with 24 h urinary protein (r=-0.228, P<0.01). 101 patients whose average level of serum complement C3 was(0.48±0.26)g/L, were treated with glucocorticoid pulse therapy as high degree of disease activity. Conclusions There is a close relationship between serum complement C3 level and the degree of disease activity in SLE. Serum complement C3 less than 0.57 g/L might be seen in SLE with severe disease activity. Key words: Lupus erythematosus, systemic; Complement system proteins; Disease activity

Differences in leukocyte differentiation molecule abundances on domestic sheep (Ovis aries) and bighorn sheep (Ovis canadensis) neutrophils identified by flow cytometry.

Although both domestic sheep (DS) and bighorn sheep (BHS) are affected by similar respiratory bacterial pathogens, experimental and field data indicate BHS are more susceptible to pneumonia. Cross-reactive monoclonal antibodies (mAbs) for use in flow cytometry (FC) are valuable reagents for interspecies comparative immune system analyses. This study describes cross-reactive mAbs that recognize leukocyte differentiation molecules (LDMs) and major histocompatibility complex antigens on DS and BHS leukocytes. Characterization of multichannel eosinophil autofluorescence in this study permitted cell-type specific gating of granulocytes for evaluating LDMs, specifically on neutrophils, by single-label FC. Evaluation of relative abundances of LDMs by flow cytometry revealed greater CD11a, CD11b, CD18 (β2 integrins) and CD 172a (SIRPα) on DS neutrophils and greater CD14 (lipopolysaccharide receptor) on BHS neutrophils. Greater CD25 (IL-2) was identified on BHS lymphocytes following Concavalin A stimulation. While DS and BHS have similar total peripheral blood leukocyte counts, BHS have proportionately more neutrophils.

Induction of systemic and mucosal immunity and maintenance of its memory against influenza A virus by nasal vaccination using a new mucosal adjuvant SF-10 derived from pulmonary surfactant in young cynomolgus monkeys

Induction of systemic and mucosal immunity and maintenance of its memory was investigated in 12 young male cynomolgus monkeys after intranasal instillation of flu vaccine using a new mucosal adjuvant SF-10 derived from pulmonary surfactant constituents. Split-product of influenza virus A/California/7/2009(H1N1)pdm hemagglutinin vaccine (HAv) at 15μg with or without SF-10 and the adjuvant alone were instilled intranasally three times every 2 weeks. SF-10-adjuvanted HAv (SF-10-HAv) elicited significantly higher HAv-specific IgG and hemagglutinin inhibition (HI) titers in serum and HAv-specific secretory IgA and its neutralizing activities in nasal washes compared with HAv antigen and SF-10 alone. Significant cross-neutralizing activities of nasal washes after the third vaccination to several other H1N1 and H3N2 strains were observed. HI titers in serum and neutralizing activities in nasal washes reached peak levels at 6 weeks after initial vaccination, then gradually decreased after 10 weeks and returned to the baseline levels at 36 weeks. A single intranasal revaccination of SF-10-HAv at 36 weeks rapidly and significantly increased both immunity in serum and nasal washes compared with naïve monkeys. Revaccination by one or two doses achieved almost maximal immunity at 2 or 4 weeks after instillation. Statistically significant adverse effects (e.g., body weight loss, elevated body temperature, nasal discharge, change in peripheral blood leukocyte and platelet counts) were not observed for 2 weeks after vaccination of SF-10-HAv, HAv or SF-10 and also during the experimental period. These results in young monkey model suggest the potential of clinical use SF-10 for intranasal flu vaccine.

A cross-sectional and follow-up study of leukopenia in tuberculosis patients: prevalence, risk factors and impact of anti-tuberculosis treatment.

To investigate the prevalence of and risk factors for leukopenia in tuberculosis patients and the impact of anti-tuberculosis regimens on the occurrence of leukopenia in newly treated tuberculosis patients. A total of 1,904 tuberculosis patients were included in the study. A cross-sectional survey of the prevalence of leukopenia was initially conducted, and then factors influencing leukopenia were identified using Logistic regression analysis. Non-treatment factors influencing peripheral blood leukocyte counts were analyzed using univariate COX proportional hazards models. Covariate analysis was used to assess the independent effect of different anti-tuberculosis regimens on peripheral blood leukocyte counts. Being female, advanced age and longer duration of previous anti-tuberculosis treatment (>6 month) were risk factors for leukopenia in tuberculosis patients, while secondary pulmonary tuberculosis, higher body mass index (BMI: 24-27.9 kg/m(2)), and higher degree of education (senior high school or above) were protective factors. Gender, vegetable consumption, drinking, pulmonary infection, other chronic diseases, and use of antibiotics were significantly associated with the development of leukopenia in patients on anti-tuberculosis treatment. In tuberculosis patients treated with anti-tuberculosis regimens not containing antibiotics, peripheral blood leukocyte levels gradually declined with the prolongation of treatment duration. In tuberculosis patients treated with anti-tuberculosis regimens containing antibiotics, peripheral blood leukocyte levels showed a declining trend. Female patients, patients at advanced age and recurrent tuberculosis patients having longer previous anti-tuberculosis treatment are high-risk populations for leukopenia. Attention should be paid to the influence of vegetable consumption and drinking, co-morbidities and use of antibiotics during anti-tuberculosis treatment.

Experimental mouse model of bronchial asthma induced by house dust mite Dermatophagoides pteronyssinus allergenic extract

The purpose of this study was to develop a mouse model of asthma (MMA) using house dust mite Dermatophagoides pteronyssinus (Der p) extract. Methods. BALB/c mice were i.p. immunized with different doses of Der p lyophilized extract three times in three week interval in the mixture with Al(OH)3. 8 weeks after the final immunization mice were challenged with Der p during five consecutive days by intranasal applications (INA) or aerosol administration (AA). All mice were divided into 5 experimental groups: group 1 was immunized with 50 ^g/mouse of Der p (in protein equivalent) in the mixture with 2 mg/mouse Al(OH)3 and challenged by INA; group 2 was immunized in the same way and challenged by AA; group 3 was immunized with 100 ^g/mouse Der p in the mixture with 2 mg/mouse of Al(OH)3 and challenged by INA; group was immunized in the same way and challenged by AA; group 5 (negative control) was immunized and challenged with saline. 24 hours after the last challenge airway hyperresponsiveness (AHR) to different concentrations of metha-choline was evaluated in all groups by whole-body plethysmography. 48 hours after the last challenge in all groups blood was collected for differential cell count, brochoalveolar lavage fluid (BALF) was sampled for the determination of inflammatory cells and lungs were removed for histological analysis. Histopathological changes in lungs (allergic inflammation) were graded according to semi-quantitative scoring system. Anti-Der p IgE, IgG1 and IgG2a antibodies in individual sera samples were detected by ELISA seven days after the last immunization and 48 hours after the challenge. Results. The levels of anti-Der p IgE antibodies in groups 1-4 before as well as after the challenge were substantially higher than that of in the group 5 (negative control). The highest level of serum Der p-specific IgE antibodies was observed in the group 2. The levels of anti-Der p IgG1 and IgG2a antibodies in the groups 1-4 during all periods of observation were higher than that of group 5 (negative control). At the same time the maximal levels of anti-Der p IgG1 and IgG2a antibodies were observed in group 3 both after immunization and after challenge. The maximum of AHR was observed in the groups 1 and 3 challenged by INA. Analysis of cell composition in BALF demonstrated significant elevated number of eosinophils in group 3 in comparison with group 5 (negative control) and other experimental groups. Regarding peripheral blood leukocyte count we observed decreasing of band neutrophils in group 4 and increasing of segmented neutrophils in groups 1 and 3 in compare to group 5 (negative control). In group 1 we found statistically significant decreasing of lymphocytes and increasing of eosinophils in compare to negative control group 5. Histological picture of general allergic inflammation in lungs as well as peribronchial and perivascular infiltration with inflammatory cells were the most noticeable (according to score system) in group 3 in comparison with negative control group and other experimental groups. Conclusion. Data obtained indicate that immunization (sensitization) of mice with Der p in a dose 100 μg/mouse together with Al(OH)3 and challenge with Der p by mean of intranasal applications is a suitable approach for modeling of mouse allergic asthma.

Human rhinovirus infection in children with acute lower respiratory tract infections

Objective To investigate the clinical and molecular epidemiologic features of human rhinovirus (HRV) in children with acute lower respiratory tract infection (ALRTI) in Shanghai by detecting HRV from bronchoalveolar lavage fluid (BALF). Methods One hundred and thirty-five BALF samples were collected from children with ALRTI at Children's Hospital of Fudan University from January to June in 2014. The specimens were tested by one-step real-time reversible transcription (RT)-polymerase chain reaction (PCR) for HRV. Gene fragments of VP4/VP2 capsid protein of the HRV genome were amplified and sequenced. The phylogenetic trees of HRV strains were drawn. Chi-square test was used for statistical analysis. Result Among these 135 samples, 6(4.4%) were HRV positive, including 4 positive for HRV-A and 2 positive for unknown genotype. The age of children with HRV positive was 8 months to 8 years, including 5 cases ≤3 years old. The nucleotide homology within 4 HRV-A samples varied 76.5%-100.0%. These 4 HRV-A sequences were respectively compared with reference strains of HRV-A, HRV-B and HRV-C in GenBank. The nucleotide homology varied 73.1%-91.7%, 57.8%-66.0% and 59.0%-65.0%, respectively. Among 135 children with ALRTI, 124 were diagnosed with ordinary pneumonia including 4 (3.2%, 4/124) HRV positive and 11 were diagnosed with severe pneumonia (with respiratory failure and/or heart failure) including 2 HRV positive. All children with HRV presented with fever and cough. Most of them (5/6) were co-infected with other pathogens. In 5 children with HRV positive, the peripheral blood leukocyte counts were less than 10×109/L, neutrophil percentages were less than 0.50, C-reactive protein was less than 8 mg/L and procalcitonin was less than 0.05 μg/L. All the children were clinically cured. Conclusions The detection of HRV in BALF samples indicates that HRV is one of the pathogens in children with ALRTI in Shanghai and HRV-A is more common. Gene fragments of VP4/VP2 capsid protein of HRV pathogenic strains display a high degree of genetic variability. Key words: Rhinovirus; Child; Acute lower respirtory infections; Bronchoalveolar lavage fluid