Routine Peripheral Blood Research Articles

Serum Immunoglobulin G Level and Neutrophils to Lymphocytes Ratio Associated with the Prognosis of Neuromyelitis Optica Spectrum Disorder.

To investigate the factors related to the prognosis of Neuromyelitis Optica Spectrum Disorder (NMOSD) in cerebrospinal fluid and peripheral blood examination. In this study, we enrolled 111 patients who were admitted to the First Affiliated Hospital of Zhengzhou University between January 2016 and January 2018 and diagnosed with NMOSD. The patients were divided into the relapse group (n = 48) and remission group (n = 67). Before treatment, all the patients underwent a routine Cerebrospinal Fluid (CSF) and peripheral blood test on the second morning of admission. The association between laboratory data and disease prognosis was evaluated. The immunoglobulin G (IgG) level in the serum showed a strong correlation with the relapse of patients, especially in the aquaporin-4-Antibody (AQP4-Ab) positive group (p < 0.01). A high level of serum IgG concentration was associated with the relapse of NMOSD, especially in the anti-AQP4 positive group. The area under the Receiver Operating Characteristic (ROC) curve of serum IgG level was 0.888 (p<0.001, 95%CI: 0.808-0.968). The ratio of Neutrophils to Lymphocytes (NLR) was associated with the disability degree of NMOSD patients in 3 years. The NLR value was a linear correlation with final Expanded Disability Status Scale (EDSS) scores; patients with a high level of NLR value presented an increased degree of disability in the following three years (R2 = 0.053, p= 0.015). The serum IgG level and NLR of first-attack patients were correlated with the prognosis of NMOSD.

EVALUATION OF MACROCYTOSIS IN PATIENTS OF BIHAR

INTRODUCTION- Macrocytosis is a relatively common nding in routine CBC and peripheral blood smear. It is divided into two groups– megaloblastic and non-megaloblastic groups based on morphological and biochemical ndings. MATERIALS AND METHODS-We conducted this study in the department of pathology, Nalanda medical college, Patna, Bihar over a period of 18 months (September 2018 to March 2020). Sixty adult patients (&gt;18 years) with macrocytosis (MCV&gt;100) were evaluated in our study. Various tests were done to establish the cause of macrocytosis. RESULT– The most common cause of macrocytosis was megaloblastic anemia due to vitamin B deciency (53.3%). Non-megaloblastic anemia 12 was caused by liver disorder (18.33%) followed by primary bone marrow disorders (10%). CONCLUSION- The commonest cause of macrocytosis is megaloblastic anemia, but there are multiple non-megaloblastic causes with different mechanism and different treatment approach. Early detection of macrocytosis and its etiology helps in proper management of the patient with better outcome.

Transfer-function-free technique for the noninvasive determination of the human arterial pressure waveform.

The estimation of central aortic blood pressure is a cardinal measurement, carrying effective physiological, and prognostic data beyond routine peripheral blood pressure. Transfer function‐based devices effectively estimate aortic systolic and diastolic blood pressure from peripheral pressure waveforms, but the reconstructed pressure waveform seems to preserve features of the peripheral waveform. We sought to develop a new method for converting the local diameter distension waveform into a pressure waveform, through an exponential function whose parameters depend on the local wave speed. The proposed method was then tested at the common carotid artery. Diameter and blood velocity waveforms were acquired via ultrasound at the right common carotid artery while simultaneously recording pressure at the left common carotid artery via tonometer in 203 people (122 men, 50 ± 18 years). The wave speed was noninvasively estimated via the lnDU‐loop method and then used to define the exponential function to convert the diameter into pressure. Noninvasive systolic and mean pressures estimated by the new technique were 3.8 ± 21.8 (p = 0.015) and 2.3 ± 9.6 mmHg (p = 0.011) higher than those obtained using tonometery. However, differences were much reduced and not significant in people >35 years (0.6 ± 18.7 and 0.8 ± 8.3 mmHg, respectively). This proof of concept study demonstrated that local wave speed, estimated from noninvasive local measurement of diameter and flow velocity, can be used to determine an exponential function that describes the relationship between local pressure and diameter. This pressure‐diameter function can then be used for the noninvasive estimation of local arterial pressure.

Investigation and analysis on the application of peripheral blood specimens for routine blood testing by laboratory physicians.

Owing to its simplicity, speed, small incision, and low cost, peripheral blood collection is widely used in routine blood testing. However, there are certain differences between the detection results of peripheral and venous blood specimens. This investigation and analysis aimed to investigate and analyze the status and opinions of laboratory physicians on the application of peripheral blood for routine blood testing. A questionnaire was used to investigate and analyze the current status of laboratory physicians with regards to routine peripheral blood testing. The content of the questionnaire included the following aspects: the proportion of peripheral blood specimens, the difficulty of detecting peripheral blood, factors affecting the results of peripheral blood specimens, the proportion of the simultaneous detection of blood test and C-reactive protein (CRP), as well as the proportion and ideals of hypersensitive CRP (hs-CRP), which included a total of 10 re-examination rules. Laboratory physicians from 410 hospitals participated in the investigation. The proportion of routine blood tests using peripheral blood specimens in the laboratory departments of tertiary hospitals was low (P=0.006). Difficulties in routine blood tests with peripheral blood specimens were dominated by insufficient blood volume (67.8%). The factors affecting the results of routine blood tests with peripheral blood specimens were dominated by the mixing method (86.6%). When abnormal results were determined by routine blood tests using peripheral blood specimens, they were retested most commonly by making slide smears (82.4%) or re-examined using the remaining peripheral blood (66.1%). The same rules for re-examination of peripheral and venous blood were applied in most medical institutions (86.1%). When the platelet count decreased in routine blood tests using peripheral blood samples, the most common measures included sample agglutination checks (88.5%), instrument alarm message checks (82.4%), and making slide smears for re-examination (73.6%). More laboratory physicians expected blood analyzers to be integrated instruments that could provide both routine blood and hs-CRP testing (80.5%). There are numerous difficulties in routine blood testing using peripheral blood, and thus, more convenient and accurate blood analysis instruments should be developed.

The Role of Neutrophil-to-Lymphocyte Ratio, Red Blood Cell Distribution Width, and Mean Platelet Volume in Predicting Febrile Seizures and Differentiating Febrile Seizure Types

AbstractThe neutrophil-to-lymphocyte ratio (NLR), red blood cell distribution width (RDW), platelet count (PLT), and mean platelet volume (MPV)/platelet ratio (MPR) are commonly known inflammatory markers measured by a routine peripheral blood test that have been studied in patients with febrile seizures (FS) and may be useful for the classification of FS types. The aim of this study was to investigate the relationship between FS and inflammatory markers including MPR, RDW, and NLR and also to determine the diagnostic ability of these parameters to identify FS by comparing patients with and without FS, and by comparing patients with FS to their FS types (simple febrile seizure or complex febrile seizure [SFS or CFS]). The study included a total of 537 children aged 6 to 60 months who presented to the emergency service with FS. The FS group was divided into two subgroups based on the type of seizure, SFS, and CFS. MPR, NLR, and RDW predicted a 1.7 (odds ratio [OR], 95% confidence interval [CI]: 1.19–2.45), 1.94 (OR, 95% CI: 1.35–2.79), and 1.8 (OR, 95% CI: 1.25–2.59) times higher risk of FS, respectively. NLR and RDW predicted a 2.64 (OR, 95% CI: 1.17–4.85) and 2.34 (OR, 95% CI: 1.14–4.44) times higher risk of recurrent SFS, respectively. In patients with CFS, NLR ≥ 1.806 had a 3.64 times (OR, 95% CI: 1.83–7.21) and RDW ≥14.55 had a 3.34 times (OR, 95% CI: 1.67–6.65) higher risk of recurrent FS. The results indicated that MPV, NLR, and RDW differentiated not only SFS from CFS but also FS from fever without seizure. The increase in RDW and NLR values and their diagnostic values in patients with recurrent FS and the diagnostic value of these parameters in predicting CFS suggest that NLR and RDW could be effective, practical, and discriminative predictors of FS.

Clinical and genetic analysis of a child with 2q37 deletion syndrome resulting from a translocation involving chromosome satellite

To carry out cyto- and molecular genetic testing for a child featuring facial dysmorphism and attention deficit and hyperactive disorder. The child was subjected to routine peripheral blood lymphocyte chromosomal karyotyping, fluorescence in situ hybridization (FISH) and single nucleotide polymorphism array (SNP-array) analyses. The child's facial dysmorphism included low-set ears, curly ear auricle, protuberance of eyebrow arch, nostril notch, short and flat philtrum and thin upper lip. SNP-array revealed that he has carried a 4.883 Mb deletion at 2q37. His chromosomal karyotype was ultimately determined as 45, XY, der(2;21) (2pter→ 2q37.3::21p13→ 21p10::20p10→ 20pter), der(20) (21qter→ 21q10::20q10→ 20qter). A rare case of 2q37 deletion syndrome involving three chromosomes was discovered. Combined use of various cyto- and molecular genetic techniques is crucial for the diagnosis of chromosomal abnormalities with complex structures.

A Case of Trisomy 9 Mosaicism Confirmed by Microarray Test

Trisomy 9 mosaicism syndrome is a rare chromosomal abnormality with a high incidence of natural abortion and perinatal death. This syndrome is characterized by intrauterine growth retardation, mental retardation, craniofacial dysmorphism including a prominent nasal bridge with a short root and a fish-shaped mouth with thin lips, skeletal abnormalities, congenital heart defects, and genital abnormalities. The incidence and severity of malformations depend on the percentage of trisomic cells in the different tissues. We report a neonate who had the characteristic features of trisomy 9 syndrome with dysmorphic features including micrognathia, microcephaly, a low-set and malformed ear, a prominent lip, and cardiac defect. No chromosomal abnormalities were detected on a routine peripheral blood chromosomal analysis; however, a chromosomal abnormality with trisomy 9 mosaicism (low-level mosaic type) was detected on genetic tests. This is thought to be due to the low proportion of trisomic cells, and for this reason, the patient in this case shows a better prognosis than four patients previously reported in Korea, they were all diagnosed by peripheral blood chromosome testing.

Neutrophil - Lymphocyte Ratio as a Reliable Predictor of Postoperative Outcome And Mortality In Delayed Cases Of Peripheral Arterial Embolism

Introduction: Peripheral arterial embolism (PAE) continues to pose a challenge to vascular surgeons and remains to be characterized by high morbidity, limb threat and mortality, often requiring urgent revascularization. Contemporary untreated arterial embolism is among the leading causes of morbidity and mortality within the vascular field. Detection of a fast and reliable biomarker for risk stratification and early post-treatment prognosis is important for those with PAE, so that individuals at high risk can be accurately treated and targeted for prevention. Complete blood count is inexpensive, comparatively routine, and is a practical laboratory test that gives important information about the patient’s formed blood contents. Routine peripheral blood counts may be useful in diagnosis and prognosis of many disorders, including peripheral vascular ischemic incidents. Neutrophil - lymphocyte ratio (NLR) has been shown to be an independent predictor of early and midterm amputation in patients with acute limb ischemia after revascularisation. Methods: Data was collected retrospectively from hospitalised patients, in the Vascular Surgery Clinic of University Hospital, Stara Zagora, Bulgaria, who had a diagnosis of PAE, from January 2010 to January 2019. Inclusion criteria were clinical data of an ischemic embolic incident of the limbs, untreated at least 12 hours from the onset of symptoms. Results: A single cutoff point was chosen to better inform clinical practice at a value of NLR 4.55, to maximize sensitivity and specificity. The majority of patients from the group Who had a NLR under 4.55 had mild symptoms. Older patients however, were significantly more likely to have an elevated NLR. The incidence of diabetes and hypertension were not significantly higher in any group. There was a higher incidence of primary major amputations in the elevated NLR group. Of the 25 deaths during the course of the study, which were specifically attributed to cardiac causes, twelve (13.8%) occurred within the elevated NLR group and thirteen (8.7%) in the low NLR group. Conclusion: NLR is an easily accessible biomarker that conveys important information about the patient’s inflammatory activity and can be easily calculated from the differential WBC count, which is routinely performed on admission and is universally available. An elevated NLR could potentially be included in addition to the most common risk factors for mortality and poor post-treatment outcome including cardiac causes, age >70 years, history of stroke or previous MI, renal failure, and smoking history (at any time).

Integrated metabolomics and network pharmacology approach to reveal immunomodulatory mechanisms of Yupingfeng granules

Systems biology is an approach that employs modern biological techniques and methods to combine the overall regulation of the body by Chinese herbal formulas with systems analysis at the molecular level. In this study, the underlying immunomodulatory mechanisms of yupingfeng granules (YPFG) were investigated based on the integration of metabolomics and network pharmacology methods. Selected routine peripheral blood indicators, body weight, and organ indices related to immunity were firstly measured in order to evaluate the effects of YPFG in cyclophosphamide-induced immunocompromised rats. Plasma metabolomics analyses were carried out by UPLC-Q-TOF-MS combined with a multivariate data analysis. Our study indicates that the potential regulatory mechanism was related to bile acid and glycerophospholipid metabolism, involving the regulation of 11 metabolites, including 8 bile acids, 1 phosphatidylserine, and 2 phosphatidylethanolamines. By means of network pharmacology, the compound-target network between potential active components of YPFG and immune dysregulation was constructed, which releated to estrogen receptor, PPAR, MAPK, PI3K-Akt, JNK signaling pathways, and ubiquitin-mediated protein degradation. The immunomodulatory effect of YPFG may be exerted through regulating lipid metabolism, then bile acid metabolism and inflammation were affected. Biological verification was also performed on cyclophosphamide-induced immunocompromised BALB/c mice. Flavonoid and saponin, two types of compounds in YPFG, were found to be the major active ingredients in the immunomodulatory effects of YPFG, and these components may regulate the abnormal metabolism of bile acids by enhancing the expression of FXR and LXRα. This work elucidated active ingredients, potential biomarkers, and mechanisms of action in the immunoregulatory effects of YPFG from the perspective of systems biology, which provides a scientific basis for its precise clinical medication.

ROUTINE PERIPHERAL BLOOD CHARACTERISTICS, IN PATIENTS WITH COVID-19 - EXPERIENCE FROM STATE COVID HOSPITAL IN SOUTH INDIA

Background: The COVID 19 pandemic is an ongoing pandemic of Corona virus disease 2019. Patients with milder symptoms have good prognosis, but severe and critical patients are difficult to treat and have high mortality rate. Many previous studies show that routine blood parameters help in predicting the progress of infectious diseases. Aims and Objectives: To analyse the peripheral blood characteristics in patients with COVID 19 and assess the efficacy, in the prognostic terms of COVID 19 patients. Material and Methods: this is a retrospective study, conducted at a state COVID hospital in South India. Results: A total of 155 cases of Covid positive patients are presented. According to severity of illness, there are 119 (76.77%) cases, categorized as mild, 24(15.48%) cases as moderate and 12 (7.74%) with severe symptoms. Conclusion: Periodic follow up of routine peripheral blood parameters, including peripheral blood film examination, help in prediction of severe COVID-19 cases.

Clinical efficacy of tocilizumab treatment in severe and critical COVID-19 patients.

BACKGROUNDThe main pathophysiological basis of coronavirus disease 2019 (COVID-19) causing respiratory failure is a cytokine storm and interleukin-6 (IL-6) is an important component of the COVID-19 cytokine storm. As a specific antagonist of IL-6, tocilizumab may block the cytokine storm of COVID-19. The Diagnosis and Treatment Guidelines of New Coronavirus Pneumonia (7th Edition) includes tocilizumab as a recommended drug for immunotherapy in severe and critical COVID-19 patients. However, the specific clinical efficacy of tocilizumab in the treatment of COVID-19 patients is worth studying.AIMTo determine the clinical efficacy of tocilizumab in inhibiting the cytokine storm in COVID-19.METHODSIn total, 19 severe and critical COVID-19 patients were enrolled in this study, and were treated with tocilizumab in Optical Valley Campus of Hubei Maternal and Child Health Care Hospital from February 20 to March 31, 2020. The imaging manifestations and clinical data before and after treatment were analyzed retrospectively, including routine peripheral venous blood tests, routine blood biochemical tests, coagulation test, C-reactive protein (CRP), IL-6, and arterial blood gas analysis.RESULTSOf the 19 patients in this group, 13 (68.4%) had significantly improved symptoms of COVID-19 (5 patients were discharged directly and 8 patients were transferred after improvement) following treatment. One case was invalid, 1 case was exacerbated, and 4 deaths (21.1%) were observed (all critical cases). The lymphocyte count, CRP, lactic acid, oxygenation index, fibrinogen (FIB) and IL-6 levels were significantly different in the improved group.CONCLUSIONTocilizumab treatment is effective against IL-6 in COVID-19 patients, but it does not completely inhibit the inflammation and cytokine storm in all patients with COVID-19.In the clinical treatment of COVID-19 patients, attention should be paid to the timing of drug administration and other adjuvant treatments.

Clinical Utility of Combined Circulating Tumor Cell and Circulating Tumor DNA Assays for Diagnosis of Primary Lung Cancer.

Although it has been suggested that circulating tumor cells (CTCs) and circulating tumor DNA (ctDNA) might be used in a complementary manner in lung cancer diagnosis, limited confirmatory data are available. In this prospective study, we evaluated the diagnostic performance of each assay separately and in combination. From March 2018 to January 2019, patients with suspected primary lung cancer, who underwent routine lung cancer work-up and peripheral blood sampling, were prospectively enrolled in the study. Epithelial cell adhesion molecule and cytokeratin served as markers of CTCs. In terms of ctDNA analysis, single-nucleotide variants were evaluated via next-generation sequencing. We analyzed 111 patients, including 99 with primary lung cancer and 12 with benign pulmonary disease. The median number of CTCs in 10 ml of blood was 3. The most frequently detected single nucleotide variants of ctDNA were TP53, CDKN2A, and EGFR. The diagnostic sensitivity of conventional tumor marker (combination of carcinoembryonic antigen/CYFRA 21-1/neuron-specific enolase) was 66.7%, while those of the ctDNA and CTC assays were 72.7% and 65.7%, respectively. The sensitivity of the CTC/ctDNA combination (95.0%) was significantly greater than those of the CTC (p<0.001), ctDNA (p<0.001), or conventional tumor marker (p<0.001) alone. Subgroup analysis revealed that the sensitivity of the combination assay was greater than those of the CTC or ctDNA assays alone, regardless of tumor stage or histopathology type. The CTC/ctDNA combination assay enhanced the sensitivity of primary lung cancer diagnosis. The combination assay strategy may be clinically useful and could enhance the early detection of lung cancer (ClinicalTrials.gov number: NCT03479099).

Construction and Identification of Acute Myeloid Leukemia NOD/SCID Mouse Model by Tail Vein Injection of THP-1 Cells

To construct NOD/SCID mouse leukemia model by using THP-1 cells. Eighteen female NOD/SCID mice aged 3 to 4 weeks were randomly divided into control group, model group A and model group B (6 in each group). Before inoculation, each mouse was intraperitoneally injected with cyclophosphamide 2 mg/(kg·d) for 2 d, and the mice in model groups were inoculated with cells within 24 h after pretreatment. The mice in model group were inoculated with THP-1 cell suspension in logarithmic growth phase by 1×107 cells/group (group A) and 5×106 cells/group (group B), the mice in the control group were injected with the same amount of normal saline in the tail vein. The general situation was observed, blood routine test and peripheral blood leukocyte classification were performed at 7, 14, 21, 28 d of inoculation before the pre-treatment, and at the time sacrifice. Before dying, tissue of mice were collected and histological examination was performed. Pilereation, droopiness and hypkinesia could be observed from d 7 and d 10 of inoculation cells in model group. Compared with the control group, the body weight of the mice in model group A and B decreased significantly after 21 days of inoculation (P<0.01), and the white blood cell counts increased significantly after 28 days of modeling (P<0.01). Among them, the above-mentioned presentation in inoculation of 1×107 group A was the most significant. Histopathological sections showed diffuse infiltration of leukemia cells in the spleen of the model group. The immunohistochemistry results indicated that the leukemia cells were positive for anti-human CD13, which confirmed the successful establishment of the model. After pretreatment with intraperitoneal injection of CTX in NOD/SCID mice, the injection of 1×107 or 5×106 THP-1 cells in tail vein of each mouse can successfully construct an acute myeloid leukemia animal model. The tumor formation is more much faster by injection of high concentration THP-1 cells.

Incidental Detection of Microfilaria in Fine Needle Aspiration Cytology- A Report of Three Cases

Lymphatic filariasis, a major global health problem, is a mosquito-borne parasitic disease, encountered mainly in the tropics and subtropics. The disease is endemic in Southeast Asia, including the Indian subcontinent. It is transmitted by mosquitoes and is caused by the filarial nematodes Wuchereria bancrofti, Brugia malayi and Brugia timori. Usually, the infected person remains asymptomatic for years together and later manifests with features and consequences of lymphatic obstruction. The adult forms of the nematodes live in human lymphatics, whereas the infective larval form, the microfilaria circulates in the peripheral blood. The nocturnal periodicity of the microfilaria makes it challenging to detect in routine peripheral blood smear examinations. Also, it is quite unusual to find the microfilaria in fine needle aspirations. However, rare cases have been reported in certain unusual sites, like lymph nodes, cutaneous swellings, bone marrow, bronchial aspirate, breast, nipple discharge, cervicovaginal smears, pleural, pericardial, ascitic fluids, ovarian cyst fluids or even thyroid. Very few cases of microfilaria in thyroid aspirates have been reported till date. Also, it is unusual to find microfilaria in lymph node and breast aspirations. Herewith, three cases of microfilariasis are reported at three uncommon sites like thyroid, lymph node and breast.

Predictive Value of Routine Peripheral Blood Biomarkers in Alzheimer's Disease.

BackgroundBiomarker screening is of major significance for the early diagnosis and prevention of Alzheimer’s disease (AD). Routine peripheral blood parameters are easy to collect and detect, making them ideal potential biomarkers. Thus, we aimed to evaluate the parameters from routine blood as potential biomarkers for AD.MethodsWe enrolled 56 AD patients, 57 mild cognitive impairment (MCI) patients, and 59 healthy elderly controls. Receiver operating characteristic (ROC) curves were used to assess the diagnostic values of routine blood biomarkers in patients with cognitive impairment.ResultsThere were significant differences in eight parameters between the groups. Logistic regression revealed that the neutrophil% (odds ratio (OR) 1.34, 95% confidence interval [CI] 1.03–1.75, p = 0.031) and neutrophil-to-lymphocyte ratio (NLR; OR 6.27, 95% CI 3.98–9.82, p = 0.003) differentiated AD patients and controls (areas under the curve [AUCs], 0.728 and 0.721) and that the NLR (OR 1.93, 95% CI 1.07–3.47, p = 0.028) and mean platelet volume (OR 1.67, 95% CI 1.04–2.70, p = 0.036) differentiated MCI patients and controls (AUCs, 0.60 and 0.638). There were no effective diagnostic biomarkers to distinguish AD from MCI.ConclusionSome routine blood biomarkers may correlate with cognitive impairment. Analysis of these biomarkers, such as the NLR, may be useful for the identification of patients with cognitive impairment.

Frequency of Lentz Bodies Inclusion in Whole Blood Erythrocytes and Expanded Buffy Coat Smears

Background: Canine distemper has been classified as highly contagious for most of domestic and wild carnivores, and the infection can be fatal. Canine distemper inclusion bodies, also denominated Lenz inclusion bodies, are large aggregates of viral nucleocapsid particles that can be form in red blood cells (RBCs), white blood cells (WBCs) and epithelial cells in many tissues during the acute phase of infection. Their presence in blood is transient and rarely encountered in light microscopy but are pathognomonic when identified in blood smears. The objective of this study was to investigate the frequency of distemper inclusions in erythrocytes according to the fraction of the sample used for blood smears. Materials, Methods &amp; Results: The study was conducted with routine blood sample provided by the Veterinary Laboratory of Clinical Analysis from the Veterinary Teaching Hospital of Universidade Federal do Rio Grande do Sul. The EDTA-K2 blood sample of a 40 days old male dog, mixed breed, no immunization records, presenting diarrhea, hyporexia, myoclonus and pustules in the abdomen, was selected. In a routine peripheral blood smear examination, several distemper inclusions were observed in the erythrocytes. From this sample, ten smears were performed using a whole blood (WB) and top erythrocyte fraction combined with buffy coat, denominated of expanded buffy coat (EBC). The EBC fraction was obtained after centrifugation of EDTA whole blood in microhematocrit tubes at 9600 x g for 5 min to obtained the packed cell volume (PCV) and buffy coat. After centrifugation, the blood cells are separated into three layers based on density: platelets (adjacent to supernatant), WBCs, and RBCs in the bottom. The PCV was measured and the microhematocrit tube was ruptured 2% below the interface between leukocytes and plasma, deposited into a plastic microtubes, homogenized and used for blood smear preparation. All smears were stained with Diff-Quick Stain. The frequency of observation of RBCs with distemper inclusions bodies was performed under optical microscopy, in the immersion objective (100x), accounting for complete fields up to a minimum of 1000 RBCs, and compared between WB and EBC. In comparison between blood smears obtained from WB and EBC, a highly significant difference (P = 0.0004) was observed in the frequency distribution of distemper inclusion. The median of frequency of RBCs with distemper inclusions in a WB smears was 12.68/1000 RBCs (10.1 - 16.1/1000 RBCs), with a coefficient of variation (CV) of 12%. Median of frequency of distemper inclusions from EBC smears was 54.23/1000 RBCs (45-77.9/1000 RBCs), CV of 18% were observed. The median frequency of inclusions found in EBC smears was 4.27 times higher than the WB smears. Discussion: Buffy coat smear providing a concentrated preparation of nucleated cells and this procedure is useful to looking for low-incidence infectious organisms or other hematologic alterations. The upper fraction of the RBC column, below the buffy coat, is composed of young RBCs. Selection of these portion, and their possible formed in the bone marrow viral replication phase, could justified the increase in the frequency of RBCs containing viral inclusions in EBC, which would also increase the sensitivity of the technique. EBC was homogenized previously to make the smears, certifying the adequate cell distribution in the slide surface without interfere with the frequency of distemper inclusion in RBCs observation. These results were confirmed with the coefficients of variation. In conclusion, distemper inclusions bodies in RBCs from EBC is a recommended diagnosis method in patients suspected of canine Distemper infection. The observation being more frequent in the EBC in comparison with WB, commonly used in veterinary hematology.

Retrospective analysis of the bone marrow received at a tertiary care hospital- A study of 160 cases

Introduction: Bone marrow examination becomes a crucial test in diagnosis of both hematological and non hematological malignancies, when the routine peripheral blood and other laboratory tests are not conclusive of a diagnosis. Aim: The aim of this study was to find the indications of bone marrow and to assess the diagnostic value. Materials and Methods: A total of 160 cases were analyzed retrospectively from January 2016 to December 2017. All the bone marrows received at the laboratory were included in the study. Result: The youngest patient was 2 years old and the oldest 80 years. Amongst 160 cases 89 (55.62%) were male and 71 (44.37%) were female. M: F ratio is 1.25:1. Leucocytosis was the most common indication of bone marrow examination followed by bicytopenia and pancytopenia. Acute leukemia was the most frequented diagnosis (13.12%) followed by idiopathic thrombocytopenic purpura (11.25%), myeloproliferative diseases (8.12%) and myeloma (8.12%). Few of the rare diagnosis included hemophagocytic syndrome, hypersplenism and aplastic anemia. Conclusion: The study concludes the bone marrow examination is crucial and important in reaching a diagnosis in most of the cryptic cases of cytopenias and unexplained leucocytosis. Keywords: Bone marrow examination, Cytopenia, Leukemia

Combination of a Cyclophosphamide-Total Body Irradiation Conditioning Regimen and Tacrolimus Plus Mycophenolate Mofetil for Gvhd Prophylaxis Is Associated with Higher HHV-6 Encephalitis Incidence after Cord Blood Transplantation

Background Human herpesvirus-6 (HHV-6) encephalitis occurs frequently through cord blood transplantation (CBT), and leads to a high risk of mortality and morbidity. Although CBT is a known risk factor for HHV-6 encephalitis, a thorough analysis of risk factors for HHV-6 encephalitis in patients receiving CBT has not been performed, and several risk factors remain unidentified. In our center, generally, either tacrolimus (Tac) plus methotrexate (MTX) (Tac/MTX) or mycophenolate mofetil (MMF) (Tac/MMF) was selected for graft-versus-host disease (GVHD) prophylaxis after CBT, at the physician's discretion. However, in recent years, Tac/MMF became the standard choice. We observed that the incidence of HHV-6 encephalitis tended to be markedly higher among patients receiving treatment with a cyclophosphamide (Cy) and total body irradiation (TBI) (Cy/TBI)-based regimen and Tac/MMF after CBT. Therefore, we aimed to ascertain whether certain conditioning regimens and GVHD prophylaxis regimens are associated with the occurrence of HHV-6 encephalitis after CBT. Methods In this retrospective study, we examined 103 patients undergoing CBT for hematological diseases at our center between 2011 and 2019. To analyze the effect of GVHD prophylaxis on the occurrence of HHV-6 encephalitis, we divided patients into two groups based on the GVHD prophylaxis regimen used: the Tac/MTX group and the Tac/MMF group. We excluded one patient who received Tac alone and one who received cyclosporine plus MTX. Finally, 101 patients were enrolled. Routine peripheral blood examinations for HHV-6 were performed once or twice a week. HHV-6 encephalitis was defined as the presence of a central nervous system dysfunction, along with a positive polymerase chain reaction result for HHV-6 DNA in the cerebrospinal fluid. HHV-6 viremia was defined as the presence of HHV-6 in the peripheral blood. Results The median age of patients was 47 years, and the median follow-up duration for surviving patients was 432 days. Conditioning regimens included myeloablative conditioning (MAC, n=64) and reduced-intensity conditioning (RIC, n=37). MAC included Cy/TBI-based regimens in 42 patients and fludarabine (Flu) plus busulfan (Bu) (Flu/Bu)-based regimens in 23, and RIC included Flu plus melphalan-based regimens in 34 patients and other regimens in 2. Sixty-four patients received Tac/MMF as GVHD prophylaxis, whereas 37 received Tac/MTX. Six patients received HLA-matched (6/6) cord blood (CB), 27 received HLA one-antigen mismatched CB, and 68 received HLA two-antigen mismatched CB. The median total nucleated cell number and CD34-positive cell number at cryopreservation were 2.2 × 107/kg and 0.7 × 105/kg, respectively. Thirteen patients (12.8%) experienced primary graft failure. Overall, the cumulative incidence of HHV-6 viremia on day 40 was 68%, and the median time from CBT to HHV-6 viremia was 23 days. The cumulative incidence of HHV-6 viremia was significantly higher in the Tac/MMF group than in the Tac/MTX group (Tac/MMF group, 75%; Tac/MTX group, 56%; p=0.03). Ten patients developed HHV-6 encephalitis. Overall, the cumulative incidence of HHV-6 encephalitis on day 60 was 10.1%. The cumulative incidence of HHV-6 encephalitis tended to be higher in the Tac/MMF group than in the Tac/MTX group (14.1% vs. 2.7%, respectively; p=0.08). Therefore, the administration of Tac/MMF as GVHD prophylaxis after CBT was identified as a probable risk factor for HHV-6 encephalitis, and we sought to identify more such risk factors, specifically among patients receiving Tac/MMF. In the analysis of Tac/MMF group, the cumulative incidence of HHV-6 encephalitis was significantly higher among patients receiving Cy/TBI-based regimens than among those receiving other regimens (35.7% vs. 8.1%, respectively; p&lt;0.01) (Fig. 1a). In addition, among the entire group of patients receiving Cy/TBI-based regimens, the cumulative incidence of HHV-6 encephalitis was significantly higher in the Tac/MMF than in the Tac/MTX group (35.7% vs. 0%, respectively; p&lt;0.01) (Fig. 1b). Conclusion Our analysis showed that patients receiving Tac/MMF as GVHD prophylaxis after CBT with a Cy/TBI-based regimen have a significantly higher risk of developing HHV-6 encephalitis. Further studies using a large cohort are required to elucidate the association between the occurrence of HHV-6 encephalitis and the use of Tac/MMF in patients undergoing CBT with a Cy/TBI-based regimen. Disclosures No relevant conflicts of interest to declare.

Circulating CD30+CD4+ T Cells Increase Before Human Immunodeficiency Virus Rebound After Analytical Antiretroviral Treatment Interruption.

Identification of nonviral markers of human immunodeficiency virus (HIV) infection that increase before viral rebound during analytical treatment interruption (ATI) may affect HIV persistence research. We previously showed that HIV ribonucleic acid (RNA) is enriched in CD30+CD4+ T cells in many individuals. Here, we studied CD30+CD4+ T-cell dynamics before ATI, during ATI (before detectable plasma RNA), and after HIV rebound. Peripheral blood mononuclear cells from 23 participants collected longitudinally from 5 Adult AIDS Clinical Trials Group studies incorporating ATI were included in this study. Flow cytometric characterization of expression of CD30 and markers of T-cell activation and exhaustion were performed along with HIV-1 RNA and deoxyribonucleic acid quantification and measurement of soluble plasma CD30 and CD30 ligand. The percentage of CD4+ T cells expressing CD30 significantly increased from pre-ATI to postinterruption time points before detectible viremia (1.65 mean relative increase, P = .005). Seventy-seven percent of participants experienced an increase in CD30+ cells before viral rebound. In contrast, there were no significant differences between pre-ATI and postinterruption pre-rebound time points in percentages of lymphocytes expressing CD69, CD38/HLA-DR, or PD-1 until after HIV recrudescence. CD30 may be a surrogate marker of early replication or viral transcriptional activity before detection by routine peripheral blood sampling.

Effect of peripheral blood lymphocyte-to-monocyte ratio on prognosis of esophageal cancer patients treated with intensity-modulated radiotherapy

Objective To observe the variation trend of the peripheral blood lymphocyte-to-monocyte ratio (LMR) during radiotherapy in patients with esophageal cancer and analyze the relationship between LMR and the radiation-induced injury, aiming to provide parameters for accurate evaluation of radiotherapy responses and clinical efficacy. Methods Clinical data of 248 eligible patients undergoing definitive radiotherapy in our department from January 2013 to December 2015, 248 were retrospectively analyzed. The routine peripheral blood examination was performed weekly before, during and at the end of radiotherapy. The absolute number of lymphocyte and monocyte was recorded to calculate the LMR. The standard classification of LMR value was conducted based on the median value of each parameter. All data including the lesion length, lesion location, clinical stage and LMR were analyzed using the Kaplan-Meier, cox and logistic regression methods, respectively. Results LMR displayed an exponential decline during radiotherapy. Univariate analysis showed that the average LMR value was the influential factor of overall survival (P=0.011) and progression-free survival (P=0.017). The mean LMR value almost exerted significant effect upon local control rate (P=0.053). No significant correlation was observed between the mean LMR value and radioactive esophagitis and pneumonitis. Stratified analysis based on the results of multivariate analysis demonstrated that patients with higher average LMR value still had longer survival. Logistic regression model revealed that the length of esophageal lesion and irradiation pattern were the influential factors of the mean LMR value. Conclusions LMR value displays an exponential decline during radiotherapy. The greater amplitude prompts the worse prognosis. The wider the irradiation field and the greater decrease in LMR exert more obvious impact on the prognosis. Key words: Lymphocyte-to-monocyte ratio; Esophageal neoplasm/intensity-modulated radiotherapy; Prognosis