Analysis of High-Risk Clinical Factors of Early Death in Secondary Hemophagocytic Lymphohistiocytosis / 中国实验血液学杂志

OBJECTIVE@#To explore the high-risk clinical factors of early death in patients with secondary hemophagocytic lymphohistiocytosis (sHLH), and further identify the clinical factors related to the rapid progression of sHLH in the short term.@*METHODS@#The clinical manifestations, laboratory examination and prognosis of sHLH patients were retrospectively analyzed. Continuous variables were grouped by median, univariate and multivariate Cox regression analysis and Kaplan-Meier survival curve were used to explore the risk factors affecting early death of sHLH. Then, a nomogram model was established with independent risk factors, Bootstrap resampling method was used for verification, and consistency index (C-index) and calibration curve were used to detect the prediction accuracy.@*RESULTS@#A total of 126 sHLH patients were enrolled, with a median age of 48.5(16-88) years, including 74 males and 52 females. Fifty-five patients (43.6%) died within 30 days, including 39 males and 16 females. Univariate regression analysis showed that lymphocyte count <0.45×109/L, platelet count (PLT) <39.5×109/L, prothrombin time (PT)≥13.3 s, activated partial thromboplastin time (APTT)≥39.7 s, albumin (ALB) <25.9 g/L, lactate dehydrogenase (LDH)≥811 U/L, creatinine (Cr) ≥67 μmol/L and procalcitonin (PCT)≥0.61 ng/ml were risk factors for death within 30 days in sHLH patients. Multivariate regression analysis showed that lymphocyte count <0.45×109/L, APTT≥39.7 s and ALB <25.9 g/L were independent risk factors for death within 30 days in sHLH patients. A nomogram model was established based on the above three risk factors, its C-index was 0.683, and the calibration chart showed good agreement between the observed and predicted values of sHLH.@*CONCLUSIONS@#Lymphopenia, prolonged APTT, and hypoalbuminemia are risk factors for early death of sHLH patients. Early identification and positive intervention are expected to reduce early mortality in sHLH patients. The nomogram model based on the above risk factors provides a method for clinicians to evaluate sHLH.

Stress-induced senescence of tumor cells by chemotherapy, good or bad? / 中国药理学通报

Traditionally it is believed that stress-induced senescence of tumor cells induced by chemotherapy is beneficial to inhibit tumor growth and progression, and has a positive effect on anti-tumor. However, with the longtime observation and clinical study on the anti-tumor effect of chemotherapy in the real world, it is found that tumor cells often show stronger proliferation and invasiveness after chemotherapy or relapse, tumor recurrence and refractoriness become very hot and tricky issues. So, it is necessary to rethink and explore the real process and potential effect of stress-induced senescence by chemotherapy. Here from the perspective of stress-induced senescence of tumor cells by chemotherapy, this study mainly analyzes and discusses the potential negative effects and clinical significance, so as to fully understand its pros and cons in the anti-tumor effect, hoping to provide some new ideas and rationale for improving traditional tumor chemotherapy, developing new anti-tumor drugs and coming out of the current dilemma of chemotherapy.

Changes and Clinical Significance of Four Parameters of NLR,LMR,RDW and PLR in Patients with Benign and Malignant Ovarian Tumors / 现代检验医学杂志

Objective To explore the changes of preoperative blood parameters of ovarian cancer patients.Methods The pop ulation that we studied consisted of 29 patients with ovarian cancer and 30 cases of ovarian benign tumors controls,calculated neutrophils and lymphocytes ratio (NLR),lymphocytes and monocytes ratio (LMR),red blood cell distribution width (RDW),platelet and lymphocyte ratio (PLR) and CA-125 levels.According to the clinical stage of ovarian cancer patients,the differences of NLR,LMR,RDW,PLR and CA 125 were analyzed in different stages.The above parameters were used to draw the ROC curve for ovarian cancer.Results The average NLR,LMR,RDW,PLR and CA 125 value in experimental and controlled were 2.49±1.04 vs 1.46±0.45,4.48±1.56 vs 6.67±1.87,175.35±62.86 vs 127.11±41.99,(12.98± 0.98)％ vs (12.48±0.76)％ and (355.9±369.73)U/ml vs (12.87±6.91)U/ml respectively.The differences were statistically significant (t =4.979.4.879.3.477,2.188,5.082,all P＜0.05).(Ⅰ + Ⅱ) stage and (Ⅲ +Ⅳ) stage of ovarian cancer patients m the above mentioned parameters mean differences were statistically significant (t=3.379,2.740,3.855,2.940,3.226,all P＜0.05).The area under ROC of NLR,LMR,RDW,PLR and CA 125 value in diagnosing ovarian cancer were 0.874,0.823,0.749,0.680 and 0.905 respectively.Conclusion Some blood routine parameters can be used to identify benign and malignant ovarian tumors,and have positive significance in the clinical staging of ovarian cancer.

The First Switched Time of PML/RARα Fusion Gene in Patients with Acute Promyelocytic Leukemia and Its Clinical Significance / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the first switched time of PML/RARα fusion gene in patients with acute promyelocytic leukemia (APL) and its clinical significance.</p><p><b>METHODS</b>sixty cases of newly diagnosed APL were enrolled in this study. They received standard remission induction, consolidation and maintenance treatments according to the clinical pathway for APL, and were followed up. During the same time the PML/RARα fusion gene mRNA expression of all cases was detected by multi-nested PCR.</p><p><b>RESULTS</b>except for 3 death cases and 1 case failed to follow-up, the PML/RARα fusion genes in the remaining 56 cases were firstly found to be negative from 24 to 381 days respectively, the mean value of the first switched time was 131 ± 90 days. There was no statistically significant difference in age, sex and risk stratification between different groups. However, the cases with L-type PML/RARα gene had shorter time compared with the patients with S-type PML/RARα gene (P = 0.032); then, for the above-mentimed 56 cases, the follow-up duration ranged from 25-1979 days (median 946 days), long-term molecular remissions had been observed in most cases, but 1 case with the first switched time of 133 days unfortunately recurred to be positive and followed by clinical relapse.</p><p><b>CONCLUSION</b>The PML/RARα fusion gene in newly diagnosed APL patients was first switched to be negative in about 4 months after treatment. The first switched time of PML/RARα fusion gene can objectively reflect the reduction of leukemia cells, and the differences among different subtypes of PML/RARα fusion gene may have some suggestions for the treatment, but without important significance for the evaluation of prognosis and recurrence for APL patients. In addition, minimal residual disease (MRD) can be dynamically monitored by detecting PML/RARα fusion gene, thus having an important clinical significance for analysis of APL recurrence.</p>

Correlation of the Desialylation of Platelets with Efficacy of the First-line Therapy for ITP / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To detect desialylation of platelets in primary immune thrombocytopenia(ITP) patients with FITC-labelled ECL and RCA-1, and compare the correlation of the desialylation level and the efficacy of first-line therapy for ITP.</p><p><b>METHODS</b>Before treatment, 48 ITP patients were selected and their levels of ECL and RCA-1 were detected with flow cytometry.</p><p><b>RESULTS</b>The desialylation level in the different efficacy groups by using the first-line therapy of corticosteroids and (or) intravenous immunoglobulin G (IVIG) had a statistically significant difference (P<0.05). The correlation analysis showed negative relation of the therapeutic efficacy with desialylation level, that is to say, the more high of desialylation level, the more poor therapeutic efficacy of the first-line therapy.</p><p><b>CONCLUSION</b>The desialylation level of platelets in ITP patients is related with the first-line therapeutic efficacy, the efficacy for patients with high desialylation level is poor, suggesting that the FcR-independent pathway exists in clearance of platelets in ITP patients. Therefore, the desialylation level of platelets may suggest the first-line therapeutic efficacy for ITP patients to a certain degree, and may be used as a potential target for the treatment of refractory ITP.</p>

New advances of epigenetic study in tumors of lymphatic system---review / 中国实验血液学杂志

Epigenetics is aimed to study the heritable changes in gene expression patterns independent of alterations in genomic DNA sequence structure, and the mechanisms of translation from genotype to phenotype. In recent years, compelling evidence gathered supports a role of epigenetic alterations in the pathogenesis of lymphatic system tumors. For example, recent data from multiple laboratories indicate that several hundred genes, involving dozens of critical molecular pathways, are epigenetically suppressed in acute lymphocytic leukemia; a panel of methylation markers can be used for additional risk stratification of chronic lymphocytic leukemia patients; based on the epigenetic profiles, the class prediction models in gray zone lymphoma can be established; the epigenetic silencing of microRNAs in multiple myeloma generally appears to have intact P53 function; epigenetic therapies have broader implication and high potential for the development of immunotherapeutic strategies and so on. In this review, the latest advances of epigenetic study and the prospect of epigenetic therapy for tumors in lymphatic system are summarized.

Fibrinogen and P-selectin expression in atherosclerosis model of Sprague Dawley rat / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Platelet P-selectin plays an important role in inflammation and contributes to thrombosis and hemostasis. Fibrinogen may take part in inflammation, thrombosis, and hemostasis via enhancement of platelet P-selectin expression. This study aimed to discover the correlation between them in atherosclerosis model of Sprague Dawley (SD) rat.</p><p><b>METHODS</b>Diet-induced atherosclerosis SD rats were adopted as experimental models. The blood from the common abdominal aorta of the rats was obtained to measure the biochemical characteristics and for the check of flow cytometry. Then the aortas were separated carefully, taken out, put into 10% (w/v) neutral formalin for later use. Then fibrinogen and P-selectin expression were detected by flow cytometry and immunohistochemistry.</p><p><b>RESULTS</b>SD rats were induced to atherosclerosis model by high fat diet and vitamin D2 injected. It was discovered that the binding of fibrinogen and the expression of P-selectin on the platelet increase in atherosclerosis model (Group H) than in that in the control group (Group Z), there were closely interrelated. High levels of fibrinogen and P-selectin express on the artery of atherosclerosis rat model.</p><p><b>CONCLUSIONS</b>Fibrinogen and P-selectin are concerned with atherosclerosis. Fibrinogen can interact with P-selectin in order to contribute to the development of atherosclerosis, high levels of fibrinogen and P-selectin can be regarded as risk factors for markers of atherosclerosis.</p>

The relationship between CD4(-) CD8(-) T cells in the peripheral blood of patients with pancreatic carcinoma and IL-4, IFN-gamma levels / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the relationship between CD4- CD8- T cells ratio and IL-4, IFN-gamma levels in the peripheral blood of patients with pancreatic carcinoma.</p><p><b>METHODS</b>Flow cytometer was used to analyze the CD4- CD8- T cells ratio in the peripheral blood of patients with pancreatic carcinoma and the IL-4, IFN-gamma levels were detected by ELISA.</p><p><b>RESULTS</b>The ratio of CD4- CD8- T cell in CD3+ T cell from 25 pancreatic carcinoma patients was (4.2 +/- 1.7)%, the ratio of CD4- CD8- T cell in CD3+ T cell from 45 healthy person was (6.3 +/- 2.6)%, there was significant deviation between the two groups (P < 0.01). The IL-4 level of 25 pancreatic carcinoma patients was (86.3 +/- 23.3) fg/L, the IL-4 level of 45 healthy person was (56.2 +/- 9.2) fg/L,there was significant deviation between the two groups (P < 0.01). The IFN-gamma level of 25 pancreatic carcinoma patients was (16.4 +/- 4.8) fg/L before operation, the IFN-gamma level of 45 healthy person was (27.4 +/- 3.8) fg/L, there was significant deviation between the two groups (P < 0.01). The ratio of CD4- CD8- T cell in pancreatic carcinoma patient after operation was higher than before operation. It could be found negative correlation between CD4- CD8- T cells ratio and IL-4 level in pancreatic carcinoma patient,it could also be found positive correlation between CD4- CD8- T cells ratio and IFN-gamma level in pancreatic carcinoma patient. In pancreatic carcinoma patient, the CD4- CD8- T cells ratio and IL-4 level was significant associated with clinical stage (P < 0.05), but no relationship with histological differentiation (P > 0.05), it could be found no relationship between IFN-gamma level and clinical stage, histological differentiation (P > 0.05).</p><p><b>CONCLUSION</b>The CD4- CD8- T cells ratio in the peripheral blood of patients is decreased,it may be participate in the carcinogenesis and development of pancreatic carcinoma by influence the IFN-gamma levels.</p>

Clinical significance of detection of T-cell subgroups in patients with aplastic anemia / 中国实验血液学杂志

The study was aimed to investigate the changes of T-cell subgroups in the peripheral blood (PB) of patients with aplastic anemia (AA) and the relationships between these changes and the pathogenesis of AA and the immunosuppressive therapeutic effects in AA, in order to provide a basis for selecting rational therapy of AA patients. T-cell subtype and the ratio of CD4+/CD8+ cell in the PB of 88 AA patients which had been diagnosed clearly and given conventional therapy or conventional therapy combined with immunotherapy were analyzed by tri-colour fluorescence-labeled monoclonal antibody and using multiparameter flow cytometry. The patients with AA were divided into normal type of ratio, inverted type of ratio, hypernormal type of ratio according to the ratio of CD4+/CD8+ cell in normal group, and then the relations of these subtype with patients' conditions and therapeutic effects were investigated. The results showed that the percentage of normal type of ratio in all patients was 39.8%, the percentage of inverted type of ratio in all patients was 44.3%, The percentage of hypernormal type of ratio in all patients was 15.9%. In the conventional therapy alone, there was no significant difference on therapeutic effects among these three immunological subtypes. In combined immunotherapy, total therapeutic efficacy of AA patients with inverted type of ratio and AA patients with immunologic abnormality (inverted type + hypernormal type) was 84.2% and 82.6% respectively, which were more than that in conventional therapy (45.5% and 42.8%) (p < 0.05). Total therapeutic efficacy in these patients was better than that in AA patients with normal type. It is concluded that significant abnormal ratios of CD4+/CD8+ exist in the majority of AA patients, abnormal ratios of CD4+/CD8+ both may be showed as increase or decrease, immunologic abnormality may play a role in pathogenesis of the patients with AA. The detection of PB T-cell subtype in patients with aplastic anemia contributes to evaluation of patients' condition and choice of rational treatment prescription, and enhancement of diagnostic level and therapeutic efficacy significantly, which is an important indicator for therapeutic strategy also.

Severe hereditary coagulation factor V deficiency caused by two novel heterozygous mutations / 中华血液学杂志

<p><b>OBJECTIVE</b>To identify gene mutations of a pedigree with inherited factor V (FV) deficiency.</p><p><b>METHODS</b>The activated partial thromboplastin time (APTT), prothrombin time (PT), FV activity (FV:C) and FV antigen (FV:Ag) tests were performed for phenotypic diagnosis. The genomic DNA was extracted from the peripheral blood of the proband and all the 25 exons and their flanks of FV gene were amplified by polymerase chain reaction (PCR). The PCR products were screened by direct sequencing and the mutations were further confirmed by restriction enzyme digestion.</p><p><b>RESULTS</b>APTT, PT, TT, FV:C, FV:Ag of the proband were 249.2 s, 46.6 s, 17.9 s, 0.1% and 1.5%, respectively. FII, FVII, FVIII, FIX, FX activities, vWF and Fg were within normal ranges. Taking the GenBank Z99572 sequence as the reference, four mutations were identified in FV gene of the proband. They were a heterozygous two bases deletion in exon 13 (2238 approximately 2239delAG) introducing a frameshift and a premature stop at codon 689, and a heterozygous missense mutation in exon 23 (G6410T) resulting in the substitution of Gly for Val at codon 2079, respectively. The proband's father and mother were heterozygous for G6410T and for 2238 approximately 2239delAG, respectively.</p><p><b>CONCLUSION</b>The severe FV deficiency of the proband is caused by a frameshift mutation of 2238 approximately 2239delAG and a missense mutation of G6410T, which haven't been identified before.</p>

Fibrinogen beta chain gene mutation contributes to one congenital afibrinogenemia / 中华血液学杂志

<p><b>OBJECTIVE</b>To identify the fibrinogen (Fg) gene mutations in a Chinese pedigree of congenital afibrinogenemia.</p><p><b>METHODS</b>The plasma Fg activity and protein of the proband and his family members were detected. Genomic DNA was isolated from the peripheral blood mononuclear cells. All the exons and exon-intron boundaries of fibrinogen gene were amplified by PCR and sequenced thereafter.</p><p><b>RESULTS</b>Two mutations, 7972 del G in FGB and T2543A in FGG, were found in the proband.</p><p><b>CONCLUSIONS</b>FGG2543 is a polymorphism site, which lead to the polymorphism of gamma144 I/K. The G deletion at base 7972 of FGB contributes to the frameshift mutation after amino acid 419, resulting in the truncated beta chain without the terminal 27 amino acids. The latter may contributes to the pathogenetic mechanisms in Chinese congenital afibrinogenemia patients. The G deletion at base 7972 of FGB is identified for the first time.</p>

Correlation of chemokines and chemokine receptors with immunological abnormality in newly diagnosed systemic lupus erythemtosus patients / 中华风湿病学杂志

Objective To explore the association of chemokines and their receptors with immunologi- cal abnormality in newly diagnosed systemic lupus erythematosus(SLE) patients.Methods The serum con- centration of MIP-1?,MIP-1?,RANTES,IFN-?IL-4 were measured by enzyme-linked immunoabsorbent assay (ELISA) in 37 newly diagnosed.SLE patients and 20 normal controls.The expression rate of CCR1, CCR3,CCR5 on CD4~+T cells were detected by flow cytometry in 18 SLE patients and 10 normal controls.Re- suits Serum MIP-1?,MIP-1?concentrations were significantly higher in SLE patients than in normal control group (P＜0.01),the concentration of MIP-1?positively correlated with MIP-1?(r=0.609,P＜0.01);the per- centage of CD4~+CCR1~+ and CD4~+CCR5~+ cell were significantly lower in newly diagnosed SLE patients than in normal control group (both P＜0.01),the percentage of CD4~+CCRI~+ cells correlated negatively with the level of serum MIP-1?and IFN-?r=-0.525,P=-0.017;r=-0.442,P=0.045);the percentage of CD4~+CCR5~+ cell corre- lated negatively with the level of serum IFN-?(r=-0.645,P=0.001);the ratios of CD4~+CCR3~+/CD4~+CCR5~+ was significantly higher in newly diagnosed SLE patients than in the normal control group (P＜0.01).Conclusion Abnormal change and interaction of chemokines and their receptors with cytokines lead to immunologic dys- function and may participate in the initiation of SLE.

Study on CD19 and CD27 of B lymphocyte subsets in peripheral blood of patients with rheumatoid arthritis and its correlation / 中华风湿病学杂志

Objective To study B lymphocyte subsets(na(?)ve B cells,memory B cells and plas- mablasts)of peripheral blood in patients with rheumatoid arthritis(RA)and its relationship with autoantibod- ies and clinical manifestation.Methods Blood samples and clinical data of 60 patients with RA were enrolled into this study.They were divided into three groups:active,inactive and refractory RA based on clinical mani- festations and 24 healthy controls were included.CD19 and CD27 of B cells in peripheral blood of RA patients and healthy controls were detected using flow cytometry at single-cell level.Frequence of na(?)ve B cells (CD19~+CD27~-),memory B cells(CD19~+CD27~(dim)),plasmablasts(CD19~+CD27~(high))and average fluorescence in- tensity of CD19 were analyzed,and their relationship with clinical manifestations and rheumatoid factor(RF), anti-typeⅡcollagen(anti-CⅡ),anti-cyclic citrullianted peptide(CCP)antibodies were investigatied.Results Frequence of na(?)ve B cells and plasmablasts in peripheral blood of patients with RA was increased compared with normal control.In contrast,memory B cells in patients with RA were decreased.The na(?)ve B cells subset in inactive and refractory RA was higher than that of healthy controls(P＜0.05),and the memory B cells subset in those groups was lower than that of healthy controls(P＜0.05).The plasmablasts in active and refractory groups of RA were higher than those of healthy controls(P＜0.05).The average fluorescence intensity of CD19 in peripheral blood in patients with RA was positively correlated with ESR,C-reactive protein(CRP),healthy assessment questionaire(HAQ),and plasmablasts was positively correlated with arthrocele index.Na(?)ve B cells,memory B cells and plasmablasts subsets had no relation with RF,anti-CⅡand anti-CCP antibodies. Conclusion B cell subsets in peripheral blood of patients with RA are significantly abnormal,characterized by expanded naive B cells and plasmablasts but diminished memory B cells.Plasmablasts are increasesd in active and refractory groups of RA,and have positive correlation with swollen joint index.B cells may play an important rote in the pathogenesis of RA.

The measurement of proportion and function of regulatory T cells in unexplained recurrent spontaneous abortion / 中华妇产科杂志

Objective To investigate the proportion and function of CD_4~+ CD_(25)~+ regulatory T cells (CD_4~+ CD_(25)~+ Tr)in unexplained recurrent spontaneous abortion(URSA).Methods(1)Proportion measurement:the proportion of CD_4~+ CD_(25)~+ Tr cells in peripheral blood was measured by double-label flow cytometric analysis.The samples were taken from 15 URSA women,15 normal non-pregnancy women and 13 normal pregnancy women.(2)Function measurement:CD_4~+ CD_(25)~+ Tr ceils and CD_4~+ CD_(25)~+ T ce]ls were extracted from peripheral blood lymphocytes by the microbeads separation.The purity of CD_4~+ CD_(25)~+ Tr cells and CD_4~+ CD_(25)~+ T cells was measured by flow cytometry.The growth inhibitory effect of CD_4~+ CD_(25)~+ Tr cells on CD_4~+ CD_(25)~+ T cells was assessed in vitro.Results The proportion of CD_4~+ CD_(25)~+ Tr cells was decreased significantly in URSA women(6.9?1.8)% than that in normal non-pregnancy women[(10.8?1.1)%] (P0.05).Conclusion The results suggest that decrease in proportion and function of CD_4~+ CD_(25)~+ Tr cells may be associated with URSA.

The Examination Method of Glycoprotein CD62P and CD63 on Platelet Membrane and Their Expression in Adults Patients with Diabetes Mellitus / 中国实验血液学杂志

To study the optimal examination method of CD62P and CD63 and investigate platelet activation in patients with diabetes mellitus (DM), whole blood labeled directly with monoclonal antibodies CD62P and CD63 and flow cytometry were used to evaluate the positive percentages and the mean fluorescence intensity of CD62P and CD63. The specimens of peripheral blood obtained from 10 healthy adults were divided into two groups. In the unfixing group, the positive percentages of CD62P and CD63 at the periods of 30, 60, 90 and 120 minutes after staining were (7.57 +/- 2.33)%, (20.50 +/- 5.70)%, (28.70 +/- 5.67)% and (36.52 +/- 6.13)%, and, (0.89 +/- 0.36)%, (1.11 +/- 0.84)%, (2.35 +/- 2.02)% and (5.43 +/- 3.66)% respectively, their respective MFI were 1.57 +/- 0.13, 1.88 +/- 0.08, 2.00 +/- 0.09 and 2.38 +/- 0.22 and 3.91 +/- 0.11, 4.07 +/- 0.16, 4.38 +/- 0.14 and 4.44 +/- 0.19. However, in fixing group with 1% paraformaldehyde, the results had not any obvious change and almost were same. Besides it, the positive percentages of CD62P and CD63 in 37 adult patients with DM were (14.11 +/- 6.68)% and (2.71 +/- 1.74)%, significantly higher than that in the normal controls. It is concluded that the CD62P and CD63 on platelet membrane were very sensitive and would be easily activated in vitro, all manipulations that includes labeling with antibody, incubation and detection using flow cytometry should be finished within 30 minutes after samples collected. While fixing by using 1% paraformaldehyde can steady the labeling compounds and effectively prevent the artificial activation of platelet, and keep the stable results within two hours after the samples labeled. In adult patients with DM, the relationship between the cardiovascular complication of diabetes and platelet activation might be existed.