The combined detection of hematological indicators is used for the differential diagnosis of colorectal cancer and benign-colorectal lesions.

OBJECTIVE: This article aims to investigate the clinical value of hemoglobin/red cell distribution width ratio (Hb/RDW), C-reactive protein/albumin ratio (CAR) and plateletcrit (PCT) combined with carcinoembryonic antigen (CEA) in colorectal cancer (CRC) auxiliary diagnosis. METHODS: We retrospectively analyzed in 718 subjects (212 with CRC, 209 with benign colorectal lesions (BCL), 111 with other cancers, and 186 healthy controls). RESULTS: The CAR, PCT, and CEA in the CRC group were higher than those in the BCL, other cancers, and the healthy control group. However, Hb/RDW in the CRC group was lower than the other three groups. Moreover, there were significant differences in Hb/RDW and CEA among different T-N-M stages (all P< 0.05). Multivariate logistic regression showed that low level of Hb/RDW and high level of CAR, CEA, PCT were risk factors for CRC, and are correlated with CRC stage. Additionally, the area under the receiver operating characteristic curve (AUC) of Hb/RDW+CEA (AUC: 0.735), CAR+CEA (AUC: 0.748), PCT+CEA (AUC: 0.807) was larger than that of Hb/RDW (AUC: 0.503), CAR (AUC: 0.614), or PCT (AUC: 0.713) alone (all P< 0.001) in distinguishing CRC from BCL. CONCLUSIONS: Hb/RDW, CAR, PCT, and CEA are independent risk factors for CRC. Hb/RDW, CAR, and PCT combined with CEA have significant value for auxiliary differential diagnosis of CRC and BCL.

Endoplasmic Reticulum Stress in the Pathogenesis of Hyperglycemia Induced by Thiamine-Responsive Megaloblastic Anemia.

BACKGROUND: Thiamine responsive megaloblastic anemia (TRMA) is a genetic disease caused by SLC19A2 gene mutation. This study aimed to preliminarily explore the relationship between endoplasmic reticulum stress (ERS)-PERK signaling pathway and the pathogenesis of hyperglycemia induced by TRMA. METHODS: Islet ß (INS.1 and ß-TC-6) and HEK293T cell line models with stable overexpression of SLC19A2 and SLC19A2 (c.1409insT) were established. The cells were divided into empty virus group (control), wild-type group (overexpressed SLC19A2), and mutation group (overexpressed SLC19A2 (c.1409insT)). Quantitative reverse transcription-polymerase chain reaction (RT-qPCR) and western blotting were used to detect the expression levels of ERS-PERK signaling pathway-related proteins, including glucose-regulated protein 78 (GRP78), protein kinase R-like ER kinase (PERK), and eukaryotic initiation factor 2 (eIF2α), activating transcription factor 4 (ATF4), and C/EBP homologous protein (CHOP) in islet ß cells. Protein localization was assessed by immunofluorescence staining. RESULTS: Compared with the control group, the mRNA expression levels of SLC19A2 in wild-type and mutant islet ß cells (INS.1 and ß-TC-6) and HEK293T cells were significantly upregulated (all p < 0.05). Compared with the control group and the wild-type group, the mRNA expression levels of GRP78, PERK, eIF2α, ATF4, and CHOP were increased (all p < 0.05) in the mutant islet ß cells; the protein expression levels of PERK, GRP78, and eIF2α were elevated (all p < 0.05). In addition, the results of immunofluorescence staining showed that SLC19A2 (c.1409insT) mutation changed the localization of the proteins in the cells. Thus, they were not located on the cell surface, but in the cytoplasm and nuclei, and protein aggregation occurred in the cytoplasm. CONCLUSIONS: 1. Islet ß and HEK293Tcell lines, stably overexpressing SLC19A2 and SLC19A2 (c.1409insT) mutations, were successfully constructed. 2. SLC19A2 (c.1409insT) mutation could raise the expression levels of ERS-PERK signaling pathway-related proteins (GRP78, PERK, eIF2α, ATF4, and CHOP), and activate apoptosis pathway. 3. SLC19A2 (c.1409insT) mutation could change the localization of proteins and produce protein aggregation in cells. It could lead to protein misfolding and ERS, which would participate in the pathological mechanism of hyperglycemia induced by TRMA.

[Genetic Analysis and Prenatal Diagnosis of a Family with Hereditary Spherocytosis Caused by a Novel Compound Heterozygous Mutation of SPTB Gene].

OBJECTIVE: To investigate the clinical and genetic characteristics of a family with hereditary spherocytosis (HS), to clarify the cause of the disease, and to provide the basis for genetic counseling and prenatal diagnosis. METHODS: The clinical data of proband and his parents were collected, and HS-related pathogenic genovariation of the proband was detected by high throughput sequencing. Suspected pathogenic mutation sites were verified by PCR-Sanger sequencing, and the fetus were conceived by a proband mother underwent prenatal diagnosis. RESULTS: Clinical manifestations of the proband showed moderate anemia, mild splenomegaly, and jaundice (an indirect increase of bilirubin). The gene detection showed that the proband showed compound heterozygous mutations of SPTB gene c. 6095T > C (p.Leu2032Pro) and c. 6224A > G (p.Glu2075Gly), which was inherited from the asymptomatic mother and father, respectively. Both mutations were detected rarely in the common population. Prenatal diagnosis revealed that the fetus inherited a mutant gene of the mother. CONCLUSION: The compound heterozygous mutations of SPTB genes c.6095T>C (p.Leu2032Pro) and c.6224A>G (p.Glu2075Gly) were the causes of the family disease, which provides a basis for family genetic counseling and prenatal diagnosis. This report is the first one found in the HGMD,1000G and EXAC database, which provides an addition to the mutation profile of the SPTB gene.

Neutrophil-to-lymphocyte ratio and lactate dehydrogenase for early diagnosis of AIDS patients with Talaromyces marneffei infection.

BACKGROUND: This study aimed to explore the value of neutrophil-to-lymphocyte ratio (NLR) in combination with routine blood tests, lactate dehydrogenase (LDH), and T-lymphocyte subsets for the early diagnosis of acquired immunodeficiency syndrome (AIDS) combined with Talaromyces marneffei (TM) infection. METHODS: A total of 166 confirmed AIDS patients were enrolled in this study. The observation group included 80 AIDS patients with TM infection, and the control group consisted of 86 AIDS patients with other complications. Regression analysis was performed to evaluate the predictive value of each index and the combination of these indexes for AIDS combined with TM infection using receiver operating characteristic (ROC) curve analysis. RESULTS: NLR and LDH were significantly higher in patients in the observation group compared with those in the control group, and the differences were statistically significant (P<0.05). There was no statistical difference in platelets, infantile granulocytes (IGM), and nucleated red blood cells (NRBC) between the 2 groups (P>0.05). The area under the operating characteristic curve (AUC) of the observed indicators were: NLR, 0.628; hemoglobin (HGB), 0.704; LDH, 0.607; lymphocyte (LYM) count, 0.744; CD4+ T lymphocyte count, 0.789; and CD8+ T lymphocyte count, 0.701. The combined AUC of multiple indicators was 0.815, with a sensitivity and specificity of 76.2% and 76.1%, respectively. CONCLUSIONS: NLR, HGB, LYM, LDH, and T lymphocyte subsets were diagnostic for early AIDS combined with TM infection , and CD4+ T lymphocytes had the best diagnostic efficacy alone.

Predictive Value of Red Blood Cell Distribution Width for 1-Year All-Cause Mortality in Critically Ill Patients with Acute Myocardial Infarction.

OBJECTIVE: Red blood cell distribution width (RDW) on admission is a prognostic factor in cardiovascular disease. This study investigated the prognostic value of the RDW measured within 24 hours before discharge (24h dRDW) on 1-year all-cause mortality in critically ill patients with acute myocardial infarction (AMI), and compared the effect of 24h dRDW in anemia and non-anemia patients. MATERIALS AND METHODS: Altogether, 4088 patients with AMI were studied retrospectively. Data from the MIMIC-III database were collected and analyzed. The Kaplan-Meier method, Cox regression models, and receiver operating characteristic (ROC) analysis were used to assess the impact of 24h dRDW on all-cause mortality in AMI patients, and a stratified analysis was performed to investigate the prognostic value of 24h dRDW in anemia and non-anemia patients. RESULTS: Of the 4088 patients, there were 704 non-survivors (17.2%). The non-survivors had a higher RDW than the survivors (p<0.001). Cox regression analysis showed that 24h dRDW had a significant independent association with 1-year all-cause mortality in critically ill patients with AMI (quintile 5 vs quintile 1, HR, 95% CI: 2.847, 2.014-4.023). The area under the ROC curve of 24h dRDW was 0.710 (95% CI, 0.689-0.730). In the stratified analysis, a significant prognostic value of 24h dRDW was found in anemia patients for 1-year all-cause mortality, but not in non-anemia patients. CONCLUSION: Elevated 24h dRDW values are significantly associated with increased hazards of all-cause mortality in critically ill patients with AMI. Significant prognostic value of 24h dRDW was found in AMI patients with anemia, but not in those without anemia.

Hereditary spherocytosis overlooked for 7 years in a pediatric patient with ß-thalassemia trait and novel compound heterozygous mutations of SPTA1 gene.

Objectives: We aimed to determine the clinical and genetic characteristics of a boy diagnosed with the ß-thalassemia trait. He also had hereditary spherocytosis (HS) that had been overlooked for 7 years. Methods: Blood samples collected from the proband and his family were assessed by laboratory tests, and next-generation sequencing (NGS) and Sanger sequencing. Results: The ß-thalassemia trait was complicated with HS in the proband. Compound heterozygous mutations of the Spectrin Alpha, Erythrocytic 1 (SPTA1) gene, c.83G > A and c.190G > A in the proband were inherited from his mother and father, respectively, and he also had the heterozygous c.126_129delCTTT mutation in the Hemoglobin Subunit Beta (HBB) gene. The c.190G > A mutation has not yet been added to the Human Gene Mutation Database (HGMD®). The heterozygous HBB c.126_129delCTTT mutation was inherited from his mother, and his older brother also had this mutation. Conclusion: Compared with other patients with either HS or ß-thalassemia, this proband with both HS and the ß-thalassemia trait had very complicated laboratory findings, which resulted in HS being overlooked for 7 years. Genetic testing is invaluable for the differential diagnosis of hereditary anemias with overlapping clinical features.

Combined Use of Mean Platelet Volume/Platelet Count Ratio and Platelet Distribution Width to Distinguish Between Patients with Nasopharyngeal Carcinoma, Those with Benign Tumors of the Nasopharynx, and Healthy Subjects.

PURPOSE: For the diagnosis of nasopharyngeal carcinoma (NPC), reliable early indicators with sensitivity and specificity should be sought. This study evaluated the effect of the combined use of mean platelet volume/platelet count ratio (MPV/PC ratio) and platelet distribution width (PDW) for differential diagnosis of NPC. In this study, MPV/PC ratio was used for the first time to diagnostically evaluate NPC. PATIENTS AND METHODS: We retrospectively analyzed various hematological indices of three subject groups (208, 185, and 162 patients with NPC, benign tumors of the nasopharynx, and healthy subjects, respectively) and evaluated the value of combined use of MPV/PC ratio and PDW for differential diagnosis of the three groups using the one-way analysis of variance. RESULTS: Comparison of laboratory variables between the three groups showed a significant difference in MPV/PC ratio and PDW (P<0.001, all). The MPV/PC ratio in the NPC group was significantly lower than the other two groups (P<0.001); MPV/PC ratio also showed a statistically significant difference in different stages (P=0.034) and serosal invasions (P<0.001) of the NPC group. Receiver operating characteristic curve (ROC) analysis showed that areas under the curve (AUC) of either patients with benign tumors of the nasopharynx (AUCMPV/PCratio+PDW: 0.708) or healthy subjects (AUCMPV/PCratio+PDW: 0.909) were larger than those of MPV/PC ratio (AUCMPV/PCratio: 0.665, 0.869, respectively) and PDW (AUCPDW:0.614, 0.716, respectively) use alone (P<0.05, all). CONCLUSION: MPV/PC ratio and PDW may be used as indexes of NPC. MPV/PC ratio combined with PDW could be considered as meaningful laboratory indexes for differential diagnosis of NPC, benign tumors of the nasopharynx, and healthy subjects. This finding could enhance the detection of NPC.

A New Compound Heterozygous Mutation Of BSCL2 In A Chinese Zhuang Ethnic Family With Congenital Generalized Lipodystrophy.

PURPOSE: This study aims to report the clinical features of an infant with CGL in a Chinese Zhuang ethnic family, whose family members were discovered to carry new pathogenic mutations in the BSCL2. PATIENTS AND METHODS: In this study, we report clinical and molecular investigations of CGL disease in a family of 4 members (parents and two sons). We used whole exome sequencing (WES) in the family to examine the genetic cause of the disease. RESULTS: The proband presented with skin pigmentation, hypertriglyceridemia and diabetes. WES identified a previously unreported compound heterozygous mutation in the BSCL2 (c.545_546insCCG heterozygous mutation and exon 3 heterozygous deletion) in the proband. His mother is a heterozygous carrier of the c.545_546insCCG mutation and his father and brother are carriers of the exon 3 heterozygous deletion. CONCLUSION: Compound heterozygous mutation of the BSCL2 (new c.545_546insCCG heterozygous mutation and new exon 3 heterozygous deletion) was detected in the proband with characteristic clinical manifestations of CGL2.

Mean platelet volume/platelet count ratio in colorectal cancer: a retrospective clinical study.

BACKGROUND: Mean platelet volume (MPV) is a marker of platelet activation. MPV and platelet count (PC) are negatively correlated, and their ratio (MPV/PC) is informative for the diagnosis of malignant tumors. However, the relationship between MPV/PC and colorectal cancer is unclear. This retrospective clinical study aimed to evaluate the diagnostic value of MPV/PC in colorectal cancer. METHODS: Hematological examinations were performed at initial diagnosis in patients with colorectal cancer (n = 186) or adenomatous polyp (n = 132) and healthy controls (n = 108). Hematological parameters evaluated included white blood cells, red blood cells, hemoglobin, neutrophils, lymphocytes, monocytes, PC, and MPV. Statistical analyses included Student's t-test, one-way ANOVA or Kruskal-Wallis H test, chi-square tests, Spearman's correlation test and receiver operating characteristic (ROC). ROC curve was used to evaluate the diagnostic values of MPV and MPV/PC in colorectal cancer. RESULTS: Among these groups, MPV was significantly lower in colorectal cancer than in adenomatous polyp (p = 0.002) and healthy controls (p < 0.001) but did not significantly differ between adenomatous polyp and healthy controls (p = 0.210). MPV/PC was lower in colorectal cancer compared with adenomatous polyp and healthy controls (p < 0.001) and in adenomatous polyp compared with healthy controls (p = 0.010). MPV did not significantly differ among colorectal cancer subgroups, while MPV/PC significantly differed between TNM stages and the presence/absence of lymph node metastasis. MPV/PC was negatively correlated with the neutrophil to lymphocyte ratio(NLR) (p = 0.002) and platelet to lymphocyte ratio(PLR) concentration (p < 0.001). In the differential diagnosis between colorectal cancer and adenomatous polyp, MPV/PC produced a larger ROC curve than MPV, NLR or PLR alone. Using MPV/PC to distinguish between colorectal cancer and controls produced a larger AUC than using MPV or NLR alone. CONCLUSIONS: MPV/PC may be useful for the diagnosis of colorectal cancer. However, further studies are warranted to include additional regions and more data, to assess the utility of MPV/PC as a novel diagnostic screening tool for colorectal cancer.

Diagnostic value of derived neutrophil-to-lymphocyte ratio in patients with ovarian cancer.

BACKGROUND: Inflammation plays an important role in the occurrence and development of cancer. Numerous studies have used the derived neutrophil-to-lymphocyte ratio (dNLR) to evaluate prognosis in many types of cancer. However, the relationship between dNLR and ovarian cancer and its value in the differential diagnosis of benign and malignant ovarian tumors remain unknown. METHODS: A total of 262 patients with ovarian cancer, 258 with benign ovarian disease, and 232 healthy controls were included in this study. dNLR was calculated using whole blood cell parameters. Receiver operating characteristic curves were generated to obtain sensitivity, specificity, and area under the ROC curve (AUC) to evaluate the diagnostic values of dNLR. RESULTS: dNLR was significantly different among the ovarian cancer, benign ovarian disease, and healthy control groups (all P < 0.001). Moreover, there were significant differences in dNLR between patients with early-stage (I and II) and advanced-stage (III and IV) disease (P < 0.001). dNLR was positively correlated with stage and carbohydrate antigen-125 in ovarian cancer. A cutoff value of dNLR ≤2.11 was diagnostic in distinguishing ovarian cancer from benign ovarian disease with AUC of 0.729 (95% confidence interval [CI], 0.689-0.767; P = 0.0001). A cutoff value of dNLR ≤1.9 was diagnostic in distinguishing ovarian cancer from healthy controls with an AUC of 0.821 (95% CI, 0.784-0.854; P = 0.0001). CONCLUSION: dNLR may be a useful indicator for distinguishing between ovarian cancer and benign ovarian disease and for identifying early and advanced ovarian cancer.

[Application of High Resolution Melting Curve Analysis in Detection of SLC4A1 Gene Mutation in Patients with Hereditary Spherocytosis].

OBJECTIVE: To investigate the feasibility and clinical significance of high resolution melting(HRM) curve analysis to detect SLC4A1 gene D38A and K56E mutations in the patients with hereditary spherocytosis(HS). METHODS: Peripheral blood was collected from 23 cases of HS for routine tests and their genomic DNA was extracted by routine technique. Specific primers of mutation sites D38A and K56E of SLC4A1 gene were designed. The HRM method was used to analyze all the samples, and then the results of HRM were verified with DNA sequencing technology. RESULTS: Among 23 specimens of HS patients, 6 cases of heterozygous mutant gene were detected by HRM technology, including 3 cases of D38A mutation and 3 cases of K56E mutation, which were confirmed by DNA sequencing. CONCLUSION: The HRM technology can correctly detect 2 common mutation sites including D38A and K56E in SLC4A1 gene in an efficient, fast, and reliable way, which not only can be used for clinical diagnosis, but also expected to be a new method for clinical researchers to define gene mutation spectrum in HS patients.

A Novel Mutation of SLC19A2 in a Chinese Zhuang Ethnic Family with Thiamine-Responsive Megaloblastic Anemia.

BACKGROUND/AIMS: Thiamine-responsive megaloblastic anemia syndrome is a rare autosomal recessive disorder resulting from mutations in SLC19A2, and is mainly characterized by megaloblastic anemia, diabetes, and progressive sensorineural hearing loss. METHODS: We study a Chinese Zhuang ethnicity family with thiamine-responsive megaloblastic anemia. The proband of the study presented with anemia and diabetes, similar to his late brother, as well as visual impairment. All clinical manifestations were corrected with thiamine (30 mg/d) supplementation for 1-3 months, except for visual impairment, which was irreversible. The presence of mutations in all exons and the flanking sequences of the SLC19A2 gene were analyzed in this family based on the proband's and his brother's clinical data. Computer analysis and prediction of the protein conformation of mutant THTR-1. The relative concentration of thiamine pyrophosphate in the proband's whole blood before and after initiation of thiamine supplement was measured by high performance liquid chromatography (HPLC). RESULTS: Gene sequencing showed a homozygous mutation in exon 6 of the SLC19A2 gene (c.1409insT) in the proband. His parents and sister were diagnosed as heterozygous carriers of the c.1409insT mutation. Computer simulation showed that the mutations caused a change in protein conformation. HPLC results suggested that the relative concentration of thiamine pyrophosphate in the proband's whole blood after thiamine supplement was significantly different (P=0.016) from that at baseline. CONCLUSIONS: This novel homozygous mutation (c.1409insT) caused the onset of thiamine-responsive megaloblastic anemia in the proband.

Molecular Genetic Mechanisms of Hereditary Spherocytosis: Current Perspectives.

With the widespread use of genetic diagnostic technologies, many novel mutations have been identified in hereditary spherocytosis (HS)-related genes, including SPTA1, SPTB, ANK1, SLC4A1, and EPB42. However, mutations in HS-related genes are dispersed and nonspecific in the diagnosis of some HS patients, indicating significant heterogeneity in the molecular deficiency of HS. It is necessary to provide the molecular and genetic characteristics of these 5 genes for clinicians to examine HS. Here, we reviewed the recent proposed molecular genetic mechanisms of HS.

Rare LPL gene missense mutation in an infant with hypertriglyceridemia.

BACKGROUND: Severe hypertriglyceridemia usually results from a combination of genetic and environmental factors and is most often attributable to mutations in the lipoprotein lipase (LPL) gene. OBJECTIVES: The aim of this study was to identify rare mutations in the LPL gene causing severe hypertriglyceridemia. METHODS: A Chinese infant who presented classical features of severe hypertriglyceridemia recruited for DNA sequencing of the LPL gene. The pathogenicity grade of the variants was defined based on the prediction of pathogenicity using in silico prediction tools. Review some studies to understand the molecular mechanisms underlying the severe hypertriglyceridemia. RESULTS: We identified a rare mutation in the LPL gene causing severe hypertriglyceridemia: a nucleotide substitution (c.836T>G) resulting in a leucine to arginine substitution at position 279 of the protein (p.Leu279Arg).The pathogenicity of the variant was predicted by in silico analysis using PolyPhen2 and SIFT prediction programs, which indicated that mutation p.Leu279Arg is probably harmful. We have also reviewed published studies concerning the molecular mechanisms underlying severe hypertriglyceridemia. A missense mutation in the 6 exon of the LPL gene is reportedly associated with LPL deficiency. CONCLUSIONS: We have here identified a rare pathogenic mutation in the LPL gene in a Chinese infant with severe hypertriglyceridemia.

Single and combined use of red cell distribution width, mean platelet volume, and cancer antigen 125 for differential diagnosis of ovarian cancer and benign ovarian tumors.

BACKGROUND: Cancer is widely believed to result from chronic inflammation, and red cell distribution width (RDW) and mean platelet volume (MPV) are considered as inflammatory markers for cancer. We investigated the values of RDW, MPV, and cancer antigen 125 (CA125), alone or in combination, for distinguishing between ovarian cancer and benign ovarian tumors. METHODS: The study included 326 patients with ovarian cancer, 290 patients with benign ovarian tumors, and 162 control subjects. Hematologic tests were performed at initial diagnosis. RESULTS: RDW was increased and MPV was decreased in the ovarian cancer group compared with the control and benign ovarian tumor groups. RDW was positively correlated and MPV was negatively correlated with cancer stage. Area under the curve (AUC) analysis for ovarian cancer versus benign ovarian tumors revealed that the specificity and sensitivity were increased for the combination of MPV and CA125 compared with either marker alone, and the specificity was increased for the combination of RDW and CA125, compared with either alone. The AUCs for RDW plus CA125 and MPV plus CA125 were significantly larger than for any of the markers alone. CONCLUSIONS: In conclusion, combinations of the markers RDW, MPV, and CA125 may improve the differential diagnosis of ovarian cancer and benign ovarian tumors.

Prevalence of hyperhomocysteinemia during routine physical examination in Guangxi Province, China and related risk factors.

BACKGROUND: Studies on homocysteine (Hcy) have mainly focused on the correlation between the homocysteine concentration and disease development. Few epidemiological investigations have been performed. This study was conducted to investigate the prevalence of hyperhomocysteinemia (HHcy) during routine physical examination in Guangxi Province, China and the correlation of serum Hcy with gender, age, serum uric acid (UA), total cholesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), and blood glucose (GLU) to provide evidence for preventing and treating HHcy. METHODS: Data of 8043 patients who underwent physical examinations at the First Affiliated Hospital of Guangxi Medical University, China from 2015 to 2016 were collected. These data included gender, age, and the serum Hcy, UA, GLU, TC, TG, HDL-C, and LDL-C concentrations. RESULTS: The overall prevalence of HHcy was 50.8% (52.3% in males, 48.1% in females). Age, UA, TC, TG, and LDL-C were significantly higher and HDL-C was significantly lower in patients with than without HHcy, regardless of gender (all P<.05). The Hcy level was positively correlated with UA, TC, TG, and LDL-C but negatively correlated with HDL-C. Gender, age, UA, TC, and TG were independent risk factors for HHcy. CONCLUSION: The prevalence of HHcy was very high during routine physical examination in Guangxi Province, China. HHcy was related to gender, age, high concentrations of UA, TC, TG, and LDL-C; and low concentrations of HDL-C. Strengthening early intervention of HHcy can reduce the risk of cardiovascular disease.

Evaluation of a Flow-Cytometric Osmotic Fragility Test for Hereditary Spherocytosis in Chinese Patients.

BACKGROUND: Osmotic fragility testing based on flow cytometry was recently introduced for the screening of hereditary spherocytosis (HS). This study was undertaken to evaluate the clinical diagnostic value of a flow-cytometric osmotic fragility test for HS. METHODS: Peripheral blood was collected from 237 subjects at the First Affiliated Hospital of Guangxi Medical University, including 56 HS patients, 86 thalassemia patients and 95 healthy controls. The samples were examined by flow-cytometric osmotic fragility test and the percentage of residual red blood cells was used to determine HS. Peripheral blood smears were performed to examine the red blood cell morphology. RESULTS: With clinical diagnosis of HS as the gold standard and the percentage of residual red blood cells <23.6% as the diagnostic threshold in the flow-cytometric osmotic fragility test, the sensitivity of the flow-cytometric osmotic fragility test for HS was 85.71% and the specificity was 97.24%. CONCLUSION: The flow-cytometric osmotic fragility test combined with a red blood cell morphology test by peripheral blood smear could be a simple, practical and accurate laboratory screening method for HS.

Evaluating γH2AX in spermatozoa from male infertility patients.

OBJECTIVE: To investigate whether γH2AX levels were different in the spermatozoa of healthy men compared with infertility patients, and to assess the possible correlations between γH2AX and conventional semen parameters and double-stranded breaks (DSBs) identified with the use of comet assay. DESIGN: Prospective study. SETTING: Clinical laboratory. PATIENT(S): Semen from 100 male infertile patients and 100 healthy sperm donors. INTERVENTION(S): Human sperm samples were analyzed in terms of World Health Organization parameters. The γH2AX levels were detected by means of flow cytometry. DSBs of sperm were detected by means of comet assay. Morphology slides were made and the sperm morphology assessed according to strict criteria. MAIN OUTCOME MEASURE(S): Conventional semen analyses, γH2AX levels in sperm, DNA DSBs in sperm, and correlations among γH2AX, conventional semen analyses, and DSBs. RESULT(S): Concentration, viability, motility, and normal sperm morphology were significantly lower in male infertility patients compared with healthy men. Also, γH2AX levels and the number of DSBs were significantly higher in the sperm of infertile subjects compared with healthy men. γH2AX levels correlated negatively with conventional semen parameters and positively with DSBs. A threshold γH2AX level of 18.55% was identified as a cutoff value to discriminate infertile subjects from fertile control subjects with a specificity of 86.0% and a sensitivity of 83.0%. The positive and negative predictive values of the 18.55% γH2AX threshold were high: 87.7% and 85.5%, respectively. CONCLUSION(S): γH2AX levels were higher in the sperm of male infertility patients than in healthy men. γH2AX levels in sperm, as evaluated with the use of flow cytometry, might be a useful biomarker for evaluating DSBs in human spermatozoa.

Comparison and evaluation of three screening tests of hereditary spherocytosis in Chinese patients.

The objective of this study is to compare and evaluate the diagnostic value of hereditary spherocytosis (HS) by three screening tests, comparing mean spherical corpuscular volume (MSCV) to mean corpuscular volume (MCV), mean corpuscular hemoglobin concentration (MCHC), and flow cytometric osmotic fragility test. Peripheral blood was collected from 237 participators diagnosed at the First Affiliated Hospital of Guangxi Medical University, including 56 hereditary spherocytosis patients, 86 thalassemia patients, and 95 healthy people. The samples were examined by three tests, and the three screening tests were evaluated by the sensitivity and specificity of tests. The sensitivity was only 41.07%, and specificity was 94.47% when using MCHC >355 g/L as diagnostic criteria. The sensitivity was 89.28%, and specificity was 96.14% when using MSCV < MCV as the optimum cutoff point. When using the residual red cell percentage <23.6% as the diagnostic threshold in flow cytometric osmotic fragility test, the sensitivity was 85.71% and the specificity was 97.24%. Flow cytometry osmotic fragility test or comparing MSCV to MCV combined with smear examination of peripheral red blood cells morphology can be a simple, practical, and accurate hereditary spherocytosis (HS) laboratory screening method.

Values of mean cell volume and mean sphered cell volume can differentiate hereditary spherocytosis and thalassemia.

OBJECTIVES: To determine whether the values of mean cell volume (MCV) and mean sphered cell volume (MSCV) can distinguish hereditary spherocytosis (HS) from thalassemia. METHODS: The MCV, MSCV, and other erythrocyte indexes were measured in totally 263 people, 57 HS patients, 109 thalassemia patients, and 107 normal control subjects. All indexes were derived from measurements obtained by the Beckman-Coulter LH 750 Hematology Analyzer. RESULTS: The MSCV was significantly lower in the HS group compared with the thalassemia group (P < 0.001), but the MCV was significantly higher in the HS group compared with the thalassemia group (P < 0.001). Among 57 patients with HS, the MCV was higher than the MSCV in 56 patients. The MCV was lower than the MSCV in one patient combined with ß-thalassemia. In the control and thalassemia groups, the MCV was lower than the MSCV. CONCLUSION: Measurements of the MCV higher than the MSCV can be considered an ideal index to distinguish rapidly HS from thalassemia.