Differential White Cell Counts Research Articles

Dynamics of differential count in dengue

Background: Dengue can occur as epidemics in India. Early diagnosis reduces mortality. Differential white cell count can aid in diagnosing and prognosticating Dengue in resource limited areas. Aim and objectives of this study were to assess patterns and utility of Differential counts in Dengue.Methods: A total of 132 serologically positive Dengue cases were analysed over the month of November 2016. Hematology data obtained from analysers and Leishman smears were tabulated and analysed.Results: The study showed lymphocytosis as the predominant pattern (65%) followed by neutropenia (30%), neutrophilia (11%), eosinophilia (5%), monocytosis (5%) and basophilia (4%). Atypical lymphocytosis ≥ 15% were noted in 65% of the cases with 83% showing Plasmacytoid lymphocytes, 8% apoptotic lymphocytes and 43% showed other atypical lymphocytes. Also, 52% of lymphocytosis and 33% of neutrophilia cases showed severe thrombocytopenia (≤ 0.5 lakhs per cu mm). Lymphocytosis was noted to be an early event but was established in later stages as seen with serology pattern association, 28% associated with NS1 antigen test (non-structural protein 1) and 42% with antibody pattern. However, neutrophilia with 60% of cases seen in antibody pattern was a late event. Plasmacytoid lymphocytosis was noted uniformly through all serology patterns in contrast with other atypical lymphocytosis which was seen mostly (48%) in antibody pattern. Apoptotic lymphocytosis was also a late event associated mainly with antibody pattern (55%).Conclusions: The Differential white cell count can be a useful supplementary test along with serology in resource limited peripheral areas. It additionally serves to drastically reduce morbidity and mortality.

Hematologic Evaluation of Painting Hall Workers in an Automobile Manufacturing Company

Background: Exposure to benzene causes several adverse effects, including decreased number of erythrocytes, leucocytes, and red blood cells, and results in aplastic anemia. The aim of this study was to investigate hematological changes due to benzene exposure in painting workers. Methods: This study was conducted on benzene-exposed and nonexposed workers in Tehran, Iran. The study sample included 40 painters exposed to different levels of benzene and 40 unexposed controls in an automobile manufacturing company. Sampling was carried out with an active pump sampler, using the NIOSH method 1501. Varian C-3800 gas chromatography-flame ionization detector (GC-FID) was used for the analysis of 40 benzene samples. Blood samples from 80 workers were collected from both the case and control groups. Hematological indices were analyzed for red blood cell, white blood cell, hematocrit, hemoglobin, platelet, and white cell differential count. Results: The average amount of benzene in painting workers was 0.775 ± 0.12 ppm. Benzene was not detected in the breathing zone of the control group. Long-term occupational exposure to benzene was lower than 1 ppm. Some blood indices, including the mean corpuscular hemoglobin concentration (MCHC) and eosinophil count, showed significant differences in comparison with the control group (P 0.05). Moreover, the platelet count was normal in workers with exposure to benzene. Conclusions: The painters showed evidence of relative exposure to benzene at a concentration below 1 ppm, causing changes in blood cells. The results of this study demonstrated that eosinophils are more sensitive than lymphocytes or neutrophils.

Prognostic Significance of Serum Inflammatory Markers in Gastric Cancer

BackgroundThe aim of this study was to assess the relative prognostic value of biomarkers to measure the systemic inflammatory response (SIR) and improve prognostic modeling in a cohort of patients undergoing potentially curative surgery for gastric adenocarcinoma. The hypothesis was that a single SIR biomarker would be associated with the most prognostic value. MethodsConsecutive 331 patients undergoing surgery for gastric cancer between 2004 and 2016 within a regional UK cancer network were identified. Serum measurements of hemoglobin, C-reactive protein, albumin, modified Glasgow Prognostic Score, and differential white cell counts were obtained before surgery, and correlated with histopathological factors (pTNM stage, differentiation, and vascular invasion) and survival. Primary outcome measures were disease-free (DFS) and overall survival (OS). ResultsConsecutive 331 patients were identified and 291 underwent potentially curative gastrectomy for adenocarcinoma. On univariable DFS analysis, female gender (p = 0.027), proximal location (p = 0.018), pT stage (p < 0.001), pN stage (p < 0.001), pTNM stage (p < 0.001), vascular invasion (p < 0.001), poor differentiation (p = 0.001), lymph node ratio (p < 0.001), R1 status (p < 0.001), platelet count (p = 0.038), and mGPS (p = 0.001) were significantly associated with poor survival. The mGPS was associated with advanced pT stage (p = 0.001), pTNM stage (p = 0.013), and poor differentiation (p = 0.030). On multivariable DFS analysis, mGPS [hazard ratio (HR) 2.51, 95% confidence interval (CI) 1.35–4.65, p = 0.011] was the only inflammatory marker to retain independent significance. Multivariable OS analysis revealed similar findings; mGPS (HR 2.75, (95% CI 1.65–4.59), p < 0.001). ConclusionmGPS is an important and only SIR-related prognostic biomarker independently associated with both DFS and OS in gastric cancer.

Immunological, Cytogenetic and Hepatoprotective Effect of Viola odorata Methanolic Extract on Methotrexate Induced Albino Male Mice

Pants used in traditional medicine contain a vast array of substances that can be used to treat chronic and infectious diseases. One of these medical plant Viola odorata which a popularly known as “Banafshah” and sweet violet in Asia and Europe respectively. It is found in high altitudes of Himalyas, Europe and throughout North America. The study was designed to assess immunological (total and differential count of white blood cells), cytogenetic (micronucleus formation) and hepatoprotective activity of methanolic extract of Viola odorata by using two doses 100 and 200mg/kg. In addition, these effects were assessed after an interaction between the two doses of the plant and methtrexte drug. Results:The results of this study indicated that Viola odorata had the ability to enhance immunity and reduced the frequency of micronucleus formation in bone marrow cells in addition to its hepatoprotective activity. Alternatively, the plant was able to counteract the damage induced by methotrexate 40 mg\kg.

Cytokine and antioxidant gene profiles from peripheral blood mononuclear cells of Pelibuey lambs after Haemonchus contortus infection.

The expression profiles of cytokines and antioxidant genes were determined from an experimental infection with H.contortus in Pelibuey lambs. The infection was followed for 34days (d) to determine the number of eggs per gram (epg) and the packed cell volume (PCV). Differential white cell counts and expression profile estimations of IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, IL-13, FCεR1A, GPX and SOD1 were determined at 0hour, 4hours, 2days and 14days post-infection (PI) in infected and control groups. Comparison of the fold change between 0 and 4-hours, 4-hours and 2-days and 2- and 14-days periods was performed. Significant differences (P<.05) between epg (>2000) and PCV (>30%) were determined after 21days and were also observed with regard to monocyte and lymphocyte cells after 2 and 7days PI. At 0hour and 14days PI, the GPX and IL-2 genes showed a 0.37- and 0.49-fold decrease in expression, respectively. In contrast, upregulation was observed at 4hours of IL-8 (2.58) and FCεR1A (2.71), at 2days for IL-4 (2.14) and IL-8 (4.02) and at 14days for IL-2 (0.41), IL-10 (2.35) and FCεR1A (2.28). The comparison between the intervals of infection showed high expression values against H.contortus infection in Pelibuey sheep after the 2nd period of PI involving a dichotomy T cells.

Single Cell Signaling Pharmacodynamics in a Phase 1b Trial of the Axl Inhibitor BGB324 in Acute Myeloid Leukemia

Axl is a receptor tyrosine kinase that has been shown to have a strong oncogenic potential in many cancer types. Overexpression and activation of Axl is found in many cancers, and is linked to increased proliferation, migration/invasion and resistance to apoptosis. Axl overexpression has been shown to be a poor prognostic marker, and recently overexpression of Axl has also been linked to the acquired resistance to chemotherapy and other anticancer therapies in many malignancies, including AML.BGB324 (BerGenBio AS) is a first-in-class highly specific small molecule inhibitor of Axl. BGB324 has been shown to be safe and well tolerated in a clinical safety trial in healthy volunteers at doses up to 1500 mg/day with a predictable PK profile and long plasma half-life, and is currently in a phase 1b clinical trial in patients with refractory/relapsed AML and MDS (BGBC003, ClinicalTrials.gov Identifier:NCT02488408; Loges S et al. J Clin Oncol 34, 2016 suppl; abstr 2561). 20 AML and 4 MDS patients have been treated at the following dose levels (loading dose/continuation dose): 400/100mg, 600/200mg and 900/300mg. Objective responses were observed in 2/4 MDS patients and 2/20 AML patients including one CR (AML). Enrollment continues to define MTD.The effect of BGB324 on intracellular signaling and the immune profile of leukemic blasts in patients treated in the clinical study was investigated using phospho-flow cytometry. Blasts were identified using surface markers (CD45low, CD66b-, CD38-, and CD117+ and/or CD34+), and the following direct and indirect downstream targets of Axl were explored: phosphorylated (p)-Akt(S473 and T308), pErk(T202/Y204), pp38(T180/Y182), pPLCγ1(Y783), pNFκB(S529), pCREB(S113) and pSTAT1(Y701), 3(Y705), 5(Y694)and 6(Y641). Preliminary analyses of blood samples from six patients show very rapid responses in signaling pathways downstream of Axl (including Akt, Erk, NFκB and PLCγ1) within hours or days of ingestion of the first dose, although the response patterns varies from patient to patient (Figure 1A). Two distinct blast populations were identified: one CD117+/CD34- and one CD117+/CD34+. In most patients the CD117+/CD34- population displayed the most extensive signaling changes during treatment, and this population also decreased during treatment with BGB324. In contrast, the CD117+/CD34+ population expanded during the course of the treatment (Figure 1B).White cell differential counts of peripheral blood from two patients treated with BGB324 for a prolonged period of time (15 weeks or more) showed a decrease in peripheral blast count, and a corresponding increase in granulocyte and monocyte counts, suggesting that Axl inhibition may push the blasts towards differentiation.The clinical trial is ongoing, and the signaling profile of leukemic blasts in blood and bone marrow of treated patients will be further examined by conventional phosphoflow cytometry and mass cytometry searching for signaling profiles with prognostic information.In conclusion, BGB324 has unique pharmacodynamic properties and molecular responses to exposure can be observed in peripheral blood leukemic blasts by phospho-flow cytometry within hours of ingestion of the first treatment dose. Further studies may establish whether single cell signal profiling can discriminate responders from non-responders and provide information about dose-response in a clinically meaningful way. [Display omitted] DisclosuresCortes:Astellas: Research Funding; Arog: Research Funding; Teva: Research Funding; Pfizer: Consultancy, Research Funding; Novartis: Consultancy, Research Funding; BMS: Consultancy, Research Funding; Ariad: Consultancy, Research Funding; Ambit: Research Funding. Heuser:Novartis: Consultancy, Research Funding; Tetralogic: Research Funding; BerGenBio: Research Funding; Karyopharm Therapeutics Inc: Research Funding; Bayer Pharma AG: Research Funding; Celgene: Honoraria; Pfizer: Research Funding. Lorens:BerGenBio AS: Employment, Equity Ownership, Research Funding. Gausdal:BerGenBio AS: Employment. Micklem:BerGenBio AS: Employment, Equity Ownership. Gjertsen:BerGenBio AS: Consultancy, Research Funding.

The Pretreatment Systemic Inflammatory Response is an Important Determinant of Poor Pathologic Response for Patients Undergoing Neoadjuvant Therapy for Rectal Cancer

BackgroundNot all patients respond equally to neoadjuvant chemoradiotherapy (nCRT), with subsequent effects on survival. The systemic inflammatory response has been shown to predict long-term outcomes in colorectal cancer. The current study examined the association between systemic inflammation and nCRT in patients with rectal cancer.MethodsBetween 1999 and 2010, patients who underwent nCRT were identified. Serum measurements of hemoglobin, C-reactive protein, albumin, modified Glasgow prognostic score (mGPS), and differential white cell counts were obtained before and after nCRT. The Rödel scoring system measured pathologic tumor regression, and magnetic resonance imaging and computed tomography determined radiologic staging.ResultsThe study included 79 patients. Of these patients, 37% were radiologically downstaged, and 44% were categorized as showing a good pathologic response (Rödel scores 3 and 4). As a validated measure of the systemic inflammatory response, mGPS (P = 0.022) was associated with a poor pathologic response to nCRT. A radiologic response was associated with a good pathologic response to treatment (P = 0.003). A binary logistic regression model identified mGPS (odds ratio [OR] 0.27; 95% confidence interval [CI] 0.07–0.96; P = 0.043) and radiologic response (OR 0.43; 95% CI 0.18–0.99; P = 0.048) as strong independent predictors of a pathologic response to treatment.ConclusionThe current study showed that a systemic inflammatory response before nCRT is associated with a poor pathologic response. Further study in a prospective controlled trial setting is warranted.

Evaluation of the performance of a point-of-care method for total and differential white blood cell count in clozapine users.

We evaluated the performance of the HemoCue WBC DIFF, a point-of-care device for total and differential white cell count, primarily to test its suitability for the mandatory white blood cell monitoring in clozapine use. Leukocyte count and 5-part differentiation was performed by the point-of-care device and by routine laboratory method in venous EDTA-blood samples from 20 clozapine users, 20 neutropenic patients, and 20 healthy volunteers. From the volunteers, also a capillary sample was drawn. Intra-assay reproducibility and drop-to-drop variation were tested. The correlation between both methods in venous samples was r > 0.95 for leukocyte, neutrophil, and lymphocyte counts. The correlation between point-of-care (capillary sample) and routine (venous sample) methods for these cells was 0.772; 0.817 and 0.798, respectively. Only for leukocyte and neutrophil counts, the intra-assay reproducibility was sufficient. The point-of-care device can be used to screen for leukocyte and neutrophil counts. Because of the relatively high measurement uncertainty and poor correlation with venous samples, we recommend to repeat the measurement with a venous sample if cell counts are in the lower reference range. In case of clozapine therapy, neutropenia can probably be excluded if high neutrophil counts are found and patients can continue their therapy.

Children’s white blood cell counts in relation to developmental exposures to methylmercury and persistent organic pollutants

BackgroundTo explore possible markers of developmental immunotoxicity, we prospectively examined 56 children to determine associations between exposures to methylmercury and persistent organic pollutants since birth and the comprehensive differential counts of white blood cells (WBC) at age 5 years. Materials and methodsExtended differential count included: neutrophils, eosinophils, basophils, lymphocytes (includingT cells, NK cells, and B cells), and monocytes. Organochlorine compounds (OCs) including polychlorinated biphenyls (PCBs) and pesticides, five perfluoroalkyl substances (PFASs), and total mercury (Hg) were measured in maternal (n=56) and children’s blood at 18 months (n=42) and 5 years (n=54). We constructed latent functions for exposures at three different ages using factor analyses and applied structural equation models adjusted for covariates. ResultsPrenatal mercury exposure was associated with depleted total WBC, especially for lymphocytes, where a one standard deviation (SD) increase in the exposure was associated with a decrease by 23% SD (95% CI: −43, −4) in the cell count. Prenatal exposure to OCs was marginally associated with decreases in neutrophil counts. In contrast, the 5-year PFASs concentrations were associated with higher basophil counts (B=46% SD, 95% CI: 13, 79). Significantly reduced subpopulations of lymphocytes such as B cells, CD4-positive T helper cells and CD4 positive recent thymic emigrants may suggest cellular immunity effects and dysregulation of T-cell mediated immunity. ConclusionDevelopmental exposure to environmental immunotoxicants appears to have different impacts on WBC counts in childhood.

Differential blood counting in fish as a non-destructive biomarker of water contamination exposure

ABSTRACTDifferential white blood cell counting was performed on blood from the fish species Oreochromis niloticus and was used as an in situ indicator of the species’ exposure to contamination. Ten young fish were collected in an area influenced by the discharge of effluents and from a fish farm (control group). The fish were anesthetized and caudal puncture was used to collect the blood. Differential white blood cell counting was performed, as well as the counting of total leukocytes and thrombocytes (in 2000 cells). Physicochemical parameters of the water from both sites were analyzed. The water from the polluted area was found to have high conductivity and low levels of dissolved oxygen, factors that indicate poor environmental quality. Fish collected from the polluted site presented higher percentages of eosinophils and monocytes and fewer thrombocytes because of exposure to pollution and hypoxic conditions. The differential white blood cell count represents a suitable biomarker of environmental health and provides a tool for biomonitoring water quality.

Bacteraemia predictive factors among general medical inpatients: a retrospective cross-sectional survey in a Japanese university hospital

ObjectivesThe precise criteria for obtaining blood cultures have not been established; they depend on the physician's judgement. We examined clinical parameters to determine predictive factors of bacteraemia and the need...

Improvement in the predictive ability of the Intermountain Mortality Risk Score by adding routinely collected laboratory tests such as albumin, bilirubin, and white cell differential count.

The Intermountain Mortality Risk Score (IMRS), a sex-specific mortality-prediction metric, has proven to be effective in various populations. IMRS is comprised of the complete blood count (CBC), basic metabolic panel (BMP), and age. Whether the addition of factors from the comprehensive metabolic panel (CMP) and white blood cell (WBC) differential count improves risk stratification is unknown. Patients with baseline complete metabolic panel (CMP) and IMRS measurements were randomly assigned (60%/40%) to independent derivation (n=84,913) and validation (n=56,584) populations. A sex-specific risk score based on IMRS methods was computed in the derivation population using adjusted multivariable regression weights from all significant and noncollinear CMP [expanded IMRS (eIMRS)] and, when available, WBC differential components (eIMRS+diff). Age averaged 67±16 years for females and 67±15 years for males. Receiver operator characteristic (ROC) c-statistics for 30-day death showed marked improvement for the eIMRS compared to the IMRS in both females [0.895 (0.882, 0.908) vs. 0.865 (0.850, 0.880)] and males [0.861 (0.847, 0.876) vs. 0.824 (0.807, 0.841)]. These results persisted for 1-year death: females [0.854 (0.847, 0.862) vs. 0.828 (0.819, 0.836)] and males [0.835 (0.826, 0.844) vs. 0.796 (0.789, 0.808)]. In addition, the eIMRS significantly improved risk reclassification. Further precision was seen when WBC differential components were included. The addition of the CMP components to the IMRS improved risk prediction. WBC differential also improved risk score predictive ability. These results suggest that the eIMRS may function even better than IMRS as a tool in patient care, risk-adjustment, and clinical research settings for predicting outcomes.

Intraepithelial lymphocytosis is a frequent finding in biopsies from ileal pouch–anal anastomoses

Following restorative proctocolectomy with an ileal pouch-anal anastomosis, the small bowel mucosa undergoes several specific histologic adaptions, which may be unrelated to the underlying disease or symptoms of pouchitis. An increase in intraepithelial lymphocytes (IELs) has not been described as part of this spectrum. Mucosal biopsies of the ileal pouch and afferent limb of 230 patients (mean age: 45.7y [18.3-74.7], gender [female/male]: 117/113) with a functioning ileal pouch-anal anastomosis (mean time since ileostomy closure: 10.8months) and associated clinically annotated outcome data were assessed for IELs/100 enterocytes. Forty-two patients (18.3%) showed an increase in IELs (≥20 IELs/100 enterocytes [range 20-39]), in pouch and/or afferent limb biopsies. Intraepithelial lymphocytosis was more commonly observed in afferent limb compared to pouch biopsies (18.8% vs 8.3%; P = .42) and in familial adenomatous polyposis compared to ulcerative colitis patients (16% vs 8%; P = 0.36), but neither difference reached statistical significance. No cases with increased IELs displayed severe villous blunting. Increased IELs were not significantly associated with age, sex, ethnicity, smoking history, time since ileostomy, use of antibiotics, biologic agents, anti-diarrheal agents or probiotics, C-reactive protein levels or differential white cell count. None of the 42 patients with increased IELs had positive celiac serology (anti-human tissue transglutaminase IgA [ELISA] with corresponding total serum IgA). Intraepithelial lymphocytosis in pouch biopsies may represent a subclinical response to an altered bacterial microenvironment. Pathologists should be aware that intraepithelial lymphocytosis is part of the spectrum of changes in pouch biopsies, and only rarely is due to celiac disease.

Evaluation of immunogenicity and protective efficacy of combination heat-killed immunogens from three entero-invasive bacteria in rabbit model

Diarrhea is a very common health problem in both developing and developed countries. Among the major entero-invasive bacteria, Shigella, Salmonella and Campylobacter cause serious problems in different geographic regions. Recently we have shown immunogenicity and protective efficacy of heat killed multi-serotype Shigella immunogen in different animal models. In our present study, we have advanced our research by preparing a combination heat-killed immunogen of three different entero-invasive bacteria, Shigella, Salmonella and Campylobacter. After three doses on 0th, 14th and 28th day of oral immunization with tri-valent heat-killed (TVHK) immunogen in rabbit model, the immunogenicity was determined by differential count of white blood cells and immunoglobulin assay at various time points. During oral immunization differential count of lymphocytes increased where as polymorphonuclear leucocytes (PMN) count decreased. Serum IgG and IgA showed significant elevation during oral immunization and remained at a detectable value upto 120 days. Protection study was performed in both, in vitro and in vivo conditions, using bacteriocidal assay and rabbit ligated ileal loop model, respectively, which conferred protection against homologous bacteria. Moreover, immunoblot assay against whole cell lysate and lipopolysaccharide exhibited significant amount of antigen-specific immunoglobulins raised against three different bacteria which proved that proteins along with lipopolysaccharides played a pivotal role in immunogenicity and protective efficacy. This trivalent heat-killed immunogen could be a low-cost, simple, oral, non-living vaccine candidate for future use against invasive diarrhea.

Clinical randomized controlled trial on the feasibility and validity of continuous blood purification during the early stage of severe burn

To observe and primarily evaluate the feasibility and validity of continuous blood purification (CBP) during the early stage of severe burn. Forty-one patients with severe burn admitted to our ward from January 2013 to July 2015, conforming to the study criteria, were divided into conventional treatment group (CT, n=21) and blood purification group (BP, n=20) according to the random number table and patient's personal consent. Patients in group CT received CT conforming to the traditional resuscitation principle for severe burn, while patients in group BP received CT and blood purification treatment in the mode of continuous venous-venous hemodiafiltration in addition up to post injury hour (PIH) 72. On post injury day (PID) 1, 2, 3, the vital signs, volume of fluid input, and volume of the urine output were observed and recorded; femoral artery blood was drawn to determine lactate, bicarbonate radical, and base excess, and oxygen index was calculated. At PIH 12, 24, 48, 72, femoral vein blood was drawn to determine white cell count, platelet count, neutrophils, creatine kinase-MB, creatine kinase, lactic dehydrogenase, aspartate transaminase (AST), alanine aminotransferase (ALT), creatinine, urea nitrogen, and blood glucose (the ratio of AST to ALT was calculated). The incidence of infection, sepsis, and multiple organ dysfunction syndrome (MODS) and the mortality of patients were recorded during 2 months after injury. Data were processed with chi-square test, analysis of variance for repeated measurement, t test and Wilcoxon test, and the values of P were adjusted by Bonferroni. The observation was completed in the 41 patients without exclusion. (1) There were no statistically significant differences in vital signs, volume of fluid input, and volume of the urine output of patients between two groups on PID 1, 2, 3 (with t values from -1.64 to 1.48, P values above 0.05). (2) Compared with that in group CT, the level of lactate of patients in group BP declined significantly on PID 2 and 3 (with Z values respectively -2.37 and -2.46, P values below 0.05). Compared with those in group CT, the levels of bicarbonate radical and base excess of patients in group BP declined significantly on PID 3 (with t values both as -2.51, P values below 0.05). The oxygen index of patients in group BP on PID 3 was (370±98) mmHg (1 mmHg=0.133 kPa), which was significantly higher than that in group CT [(305±81) mmHg, t=2.27, P<0.05]. (3) There were no statistically significant differences in white cell count, platelet count, neutrophils, creatine kinase, lactic dehydrogenase, AST, ALT, and AST to ALT ratio of patients between two groups at PIH 12, 24, 48, 72 (with t values from -1.47 to 1.19, Z values from -1.58 to -0.03, P values above 0.05). At PIH 24, 48, 72, the levels of creatine kinase-MB and blood glucose of patients in group BP were respectively (81±43), (55±34), (58±40) U/L and (7.9±2.0), (6.7±0.9), (6.9±1.8) mmol/L, which were significantly lower than those in group CT [(179±184), (124±71), (103±57) U/L and (10.1±3.8), (9.1±2.4), (8.8±4.1) mmol/L, with Z values from -3.73 to -2.02, P<0.05 or P<0.01]. Compared with those of patients in group CT, creatinine at PIH 48 and urea nitrogen at PIH 24, 48, 72 were obviously lower in group BP (with t values from -4.23 to -2.44, P<0.05 or P<0.01). (4) During the two months after injury, the infection rate of patients in group BP was 60.0% (12/20), which was significantly lower than that in group CT [95.2% (20/21), χ(2)=5.51, P<0.05]. The incidence of sepsis and MODS and the mortality of patients in group BP were all lower than those in group CT, but there were no statistically significant differences (with χ(2) values from 0.22 to 2.93, P values above 0.05). Conducting CBP in the early stage of severe burn is safe and feasible, which does not obviously affect the vital signs, volumes of fluid input and urine output, or platelet count of patients, additionally, it could help protect the function of vital organs, eliminate stress hyperglycemia, and reduce infection rate. Clinical trial registration Chinese Clinical Trial Registry, ChiCTR-TRC-12002616.

Mismatch repair status in patients with primary operable colorectal cancer: associations with the local and systemic tumour environment.

Background:Mismatch repair-deficient (dMMR) colorectal cancer (CRC) is associated with a conspicuous local immune infiltrate; however, its relationship with systemic inflammatory responses remains to be determined. The present study aims to examine the relationships and prognostic value of assessment of the local and systemic environment in the context of MMR status in patients with CRC.Methods:The relationship between MMR status, determined using immunohistochemistry, and the local inflammatory cell infiltrate, differential white cell count, neutrophil : platelet score (NPS), neutrophil : lymphocyte ratio and modified Glasgow Prognostic Score (mGPS), and cancer-specific survival was examined in 228 patients undergoing resection of stage I–III CRC.Results:Thirty-five patients (15%) had dMMR CRC. Mismatch repair deficiency was associated with a higher density of CD3+, CD8+ and CD45R0+ T lymphocytes within the cancer cell nests and an elevated mGPS (mGPS2: 23% vs 9%, P=0.007) and NPS (NPS2: 19% vs 3%, P=0.001). CD3+ density (P<0.001), mGPS (P=0.01) and NPS (P=0.042) were associated with survival independent of MMR status (P=0.367) and stratified 5-year survival of patients with MMR-competent CRC from 94% to 67%, 83% to 46% and 78% to 60% respectively.Conclusions:Mismatch repair deficiency was associated with local and systemic environments, and in comparison with their assessment, dMMR had relatively poor prognostic value in patients with primary operable CRC. In addition to MMR status, local and systemic inflammatory responses should be assessed in these patients.

Evaluation of the accuracy of the CellaVision™ DM96 in a high HIV-prevalence population in South Africa.

IntroductionThe CellaVision™ DM96 (DM96) is a digital microscopy system which performs well in developed countries. However, to date it has not been evaluated in Africa, where the pathology spectrum encountered is very different. In particular, its utility in a setting with high HIV prevalence has not been assessed, which is of interest because of the morphological aberrations often seen in HIV-positive patients.ObjectivesThis study aimed to evaluate the accuracy of the DM96 in a South African laboratory, with emphasis on its performance in samples collected from HIV-positive patients.MethodsA total of 149 samples submitted for a routine differential white cell count in 2012 and 2013 at the Chris Hani Baragwanath Academic Hospital in Johannesburg, South Africa were included, of which 79 (53.0%) were collected from HIV-positive patients. Results of DM96 analysis pre- and post-classification were compared with a manual differential white cell count and the impact of HIV infection and other variables of interest were assessed.ResultsPre- and post-classification accuracies were similar to those reported in developed countries. Reclassification was required in 16% of cells, with particularly high misclassification rates for eosinophils (31.7%), blasts (33.7%) and basophils (93.5%). Multivariate analysis revealed a significant relationship between the number of misclassified cells and both the white cell count (p = 0.035) and the presence of malignant cells in the blood (p = 0.049), but not with any other variables analysed, including HIV status.ConclusionThe DM96 exhibited acceptable accuracy in this South African laboratory, which was not impacted by HIV infection. However, as it does not eliminate the need for experienced morphologists, its cost may be unjustifiable in a resource-constrained setting.

Assessment of inhaled acute ammonia-induced lung injury in rats

This study examined acute toxicity and lung injury following inhalation exposure to ammonia. Male Sprague-Dawley rats (300–350 g) were exposed to 9000, 20 000, 23 000, 26 000, 30 000 or 35 000 ppm of ammonia for 20 min in a custom head-out exposure system. The exposure atmosphere, which attained steady state within 3 min for all ammonia concentrations, was monitored and verified using a Fourier transform infrared spectroscopy (FTIR) gas analyzer. Animals exposed to ammonia resulted in dose-dependent increases in observed signs of intoxication, including increased chewing and licking, ocular irritation, salivation, lacrimation, oronasal secretion and labored breathing. The LCt50 of ammonia within this head-out inhalation exposure model was determined by probit analysis to be 23 672 ppm (16 489 mg/m3) for the 20 min exposure in male rats. Exposure to 20 000 or 23 000 ppm of ammonia resulted in significant body weight loss 24-h post-exposure. Lung edema increased in all ammonia-exposed animal groups and was significant following exposure to 9000 ppm. Bronchoalveolar fluid (BALF) protein concentrations significantly increased following exposure to 20 000 or 23 000 ppm of ammonia in comparison to controls. BAL cell (BALC) death and total cell counts increased in animals exposed to 20 000 or 23 000 ppm of ammonia in comparison to controls. Differential cell counts of white blood cells, neutrophils and platelets from blood and BALF were significantly increased following exposure to 23 000 ppm of ammonia. The following studies describe the validation of a head-out inhalation exposure model for the determination of acute ammonia-induced toxicity; this model will be used for the development and evaluation of potential therapies that provide protection against respiratory and systemic toxicological effects.

Blood smear analyzer for white blood cell counting: A hybrid microscopic image analyzing technique

Total count and differential count of leukocytes or white blood cells (WBC) in blood samples are very important pathological factors for diagnosing a disease. There are not enough pathological infrastructures in the remote places of India and other developing countries. The objective of this work is to design a system, compatible with telemedicine, for automatic calculation of the total count and differential count of WBC from the blood smear slides. Hemocytometer based WBC counting provides more accurate result than manual counting, but hemocytometer preparation process needs expertise. As this device is targeted for remote places, blood smear technique is adopted to reduce the overhead of the operator. In the proposed system, microscopic images of blood smear sample are processed to highlight the WBC for segmentation. Region segmentation procedure involves background scaling and redundant region elimination from the region set. After segmentation, the more accurate region boundary is restored by using gradient based region growing with neighbourhood influence. Individual regions are separately classified on the basis of shape, size, color and texture features independently using different fuzzy and non-fuzzy techniques. A final decision is taken by combining these classification results, which is a kind of hybridization. A set of rules has been generated for making final classification decision based on outputs from various classifiers. The sensitivity and specificity of the system are found to be 96.4% and 79.6%, respectively on a database of 150 blood smear slides collected from different health centres of Kolkata Municipal Corporation, Kolkata, India.

Antiinflammatory Effect of N-Acetylcysteine Combined with Exogenous Surfactant in Meconium-Induced Lung Injury.

Neonatal meconium aspiration syndrome (MAS) can be treated by exogenous surfactant (S). However, aspirated meconium initiates local inflammation and oxidation which may inactivate surfactant and reduce its action. This experimental study estimated whether combined use of surfactant and the antioxidant N-acetylcysteine (NAC) can enhance effectiveness of therapy. Meconium-instilled rabbits were non-treated (M), treated with monotherapies (M + S, M + NAC), combined therapy (M + S + NAC), or received saline instead of meconium (controls, C). Surfactant therapy consisted of two lung lavages (BAL) with diluted Curosurf (5mg phospholipids/ml, 10ml/kg) followed by undiluted Curosurf (100mg phospholipids/kg). N-acetylcysteine (Acc Injekt, 10mg/kg) was given intravenously in M + S + NAC group 10min after surfactant therapy. Animals were oxygen-ventilated for additional 5h. Then, differential white cell count in the blood (WBC) was determined. Left lung was saline-lavaged and differential cell count in BAL was determined. In right lung tissue, wet/dry weight ratio, oxidation markers (TBARS, 3NT) and interleukines (IL-2, IL-6, IL-13, and TNFα) using ELISA and RT-PCR were estimated. Combined S + NAC therapy significantly decreased W/D ratio, TBARS, 3NT, and IL, whereas the effect of monotherapies (either S or NAC) was less obvious. In conclusion, addition of NAC to surfactant treatment may enhance the therapeutic outcome in MAS.