Identification of Weak D Type 1 in Rh Blood Group System and Discussion of Transfusion Strategy / 中国实验血液学杂志

OBJECTIVE@#To investigate the molecular mechanism of one patient with abnormal serological phenotype in RhD and discuss the transfusion strategy.@*METHODS@#The RhD variant sample was screened from a patient with IgM type anti-D antibody and further determined by three different sources of anti-D antibodies. Ten exons and the adjacent introns of the RHD gene were amplified, purified and sequenced. RhCE phenotypes and RHCE genotypes were detected.@*RESULTS@#The patient with Rh variant showed abnormal results of serological tests. The RHD gene sequence analysis showed that the RHD*01W.01 with a variation (c.809T>G, p.Val270Gly) in exon 6 of the RHD gene was found in the patient. The RhCE phenotype was CcEe. The genotyping results of RHCE were consistent with the serological typing results.@*CONCLUSION@#The Rh variant of the patient is RHD*01W.01, these findings indicate that RhD variants should be analyzed by molecular assays for the sake of safe transfusion.

Different Subtypes Caused by c.721C>T Substitution in the Exon 7 of ABO Gene / 中国实验血液学杂志

OBJECTIVE@#To analyze the different subtypes caused by c.721C>T substitution in the exon 7 of the ABO gene, and to investigate the related molecular mechanism of different antigens expression.@*METHODS@#ABO subtypes in 7 samples were identified by standard serological methods. The exons 6, 7, and adjacent intron of ABO gene were amplified by Polymerase Chain Reaction (PCR), and the PCR products were analyzed by direct DNA sequencing and cloning sequencing.@*RESULTS@#ABO subtypes phenotypes were A@*CONCLUSION@#c.721C>T substitution in the ABO gene causes p.Arg241Trp exchange resulting in the decreasing of GTA or GTB activities and weaker antigen expression. O.01.07 is a null allele which cannot form a functional catalytic enzyme has no effect on A

Study on the Identification of Samples Mistyped as O Phenotype and the Strategy of Safe Blood Transfusion / 中国实验血液学杂志

OBJECTIVE@#To investigate the indentification method of samples mistyped as O phenotype and to explore the precision transfusion strategy.@*METHODS@#The blood samples from donors and patients admitted in our center from 2018 to 2019 was collected. The samples with O phenotype suspected subtypes were further determined by tube test, adsorption-elution test, etc. Molecular testing was used to sequence the related blood type genes of the subjects.@*RESULTS@#Among 14 subjects misjudged as O, 11 different genotypes were identified, in which 3 blood donors were Ael02/O02, Bel03/O02, and one para-Bombay with B101/O02 (FUT1: h3h3; FUT2: Se@*CONCLUSION@#The phenotypes of Ael, Bel, Aw and para-Bombay subtypes are easily misjudged as type O. Molecular technology is helpful to identify the genotype of subtypes, and the corresponding transfusion strategies could be reasonably performed.

Identification of a Rare Ael05/B101 Subtype and Selection of Blood Transfusion Strategy / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore the mechanism of ABO discrepancy in a patient by ABO genotyping and the reasonable blood transfusion strategy.</p><p><b>METHODS</b>Routine serological test was carried out to analyze ABO blood group. The presence of the blood group determinants on the red blood cells were determined by adsorption-elution test. Exons 1-7 and adjacent introns of the ABO gene were amplified by PCR and sequenced.</p><p><b>RESULTS</b>The patient showed ABO forward and reverse typing discrepancy. ABO forward typing defined as B, however, the reverse typing indicated that the patient was AB subtype. Absorption-elution test confirmed weak A antigens on the patient's red blood cells. The ABO gene sequencing showed a T>C exchange at position in exon 7 which resulted in a isoleucine to threonine substitution at codon 256. The ABO blood group genotype was ABO*Ael05/B101.</p><p><b>CONCLUSION</b>The 767 T>C substitution in the gene of α-1,3-N-acetyl galactose is the molecular mechanism leading to the decrease expression of A antigen of the Ael05 subtype.</p>

Polymorphisms of RHCE gene among serologic RhD negative donors of Han population in Chinese Jiangsu area / 中国实验血液学杂志

This study was purposed to investigate the RHCE gene polymorphisms in Chinese Jiangsu Han blood donors with and without RHD gene among serologic RhD negative population. PCR with sequence-specific primers (PCR-SSP) was used to detect RHCE genotype in 337 serologic RhD negative, RHD gene positive donors. The RHD gene-specific polymorphisms were also determined by PCR-SSP in these donors. The results showed that among 337 serologic RhD negative, RHD gene positive donor 20 were RHCE*C/C, 62 RHCE*C/c, 24 RHCE*c/c, 25 RHCE*E/e, and 81 RHCE*e/e; the allele frequencies for RHCE*C and RHCE*c were 0.4811 and 0.5189, respectively; and for RHCE*E and RHCE*e 0.1179 and 0.8821. Among 231 RHD gene negative donors, 3 were RHCE*C/C, 34 RHCE*C/c, 194 RHCE*c/c, 15 RHCE*E/e, 216 RHCE*e/e; the allele frequencies for RHCE*C and RHCE*c were 0.0866 and 0.9134, respectively; and allele frequencies for RHCE*E and RHCE*e were 0.0325 and 0.9675, respectively. It is concluded that the most prevalent allele of RHCE gene was RHCE*ccee in Chinese Han Jiangsu RHD gene negative population. There is statistical difference in RHCE genotype distribution among serologic RhD negative population with and without RHD gene.

Feasibility of volume perfusion CT (VPCT) imaging in antiangiogenic treatment of rabbit VX2 soft-tissue tumor / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To explore the feasibility of volume perfusion CT imaging to dynamically monitor and evaluate the response of rabbit VX2 soft-tissue tumor to antiangiogenic treatment.</p><p><b>METHODS</b>To establish an experimental animal model of VX2 soft tissue tumor on 20 New Zealand white rabbits. Twenty rabbits were randomly divided into 2 groups. The therapy group was treated with recombinant human endostatin (3 mg·kg⁻¹·d⁻¹) for 7 days, and the control group received saline in the same dose only. Four times of CT volume perfusion scan were performed before treatment and on the second, forth, seventh days of treatment, respectively. The value of blood flow (BF), blood volume (BV), mean transit time (MTT), and permeability (PMB) in the VX2 tumors were measured after scanning. The microvessel density (MVD) and expression of vascular endothelial growth factor (VEGF) in the tumors were determined using immunohistochemical staining.</p><p><b>RESULTS</b>The tumor volume of the therapy group was (1.36 ± 0.73) cm³ on the forth day of treatment and (1.69 ± 0.68) cm³ on the seventh day of the treatment. The tumor volume of the control group was (2.35 ± 0.62) cm³ on the fourth day of treatment and (3.87 ± 0.93) cm³ on the seventh day of the treatment (P < 0.05). On the seventh day of treatment, tumor necrosis ratio of the therapy group and the control group was (25.58 ± 5.51)% and (42.93 ± 4.34)%, respectively (P < 0.05). Comparing the perfusion parameters between the two groups on the same day, and the second, forth, seventh days of treatment, the value of PMB of the therapy group was (70.36 ± 23.46) ml·100 ml⁻¹·min⁻¹, (79.64 ± 13.68) ml·100 ml⁻¹·min⁻¹ and (84.76 ± 3.55) ml·100 ml⁻¹·min⁻¹, respectively, and that in the control group was (26.61 ± 6.47) ml·100 ml⁻¹·min⁻¹, (33.74 ± 16.47) ml·100 ml⁻¹·min⁻¹ and (30.47 ± 10.64) ml·100 ml⁻¹·min⁻¹, respectively (P < 0.05). The value of BF in the therapy group and control group was (71.19 ± 12.21) ml·100 ml⁻¹·min⁻¹ and (43.56 ± 12.21) ml·100 ml⁻¹·min⁻¹, respectively, on the seventh day of treatment (P < 0.05). The parameters on different days in the same group were compared. In the control group, the value of BF on the seventh day of treatment was significantly lower than that before and on the second and forth days of treatment (P < 0.05). However, in the therapy group, the value of PMB on the second, forth, and seventh days of treatment was significantly higher than that before treatment (P < 0.05). MVD of tumor in the control group was increased gradually, whereas increased on the first day and then decreased more in the therapy group. The VEGF expressions did not differ significantly between the experimental and control groups.</p><p><b>CONCLUSIONS</b>Volume perfusion CT is helpful to quantify the tumor perfusion and evaluate the functional changes of tumor vasculature, and then evaluate the early therapeutic effect of antiangiogenic treatment.</p>

Analysis of cavitation of advanced NSCLC treated by rh-endostatin combined with NP chemotherapy / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To study the significance of intra-tumoral cavitation in the patients with advanced NSCLC treated by rh-endostatin plus NP chemotherapy.</p><p><b>METHODS</b>Fifty-seven patients with advanced NSCLC were randomly assigned to receive chemotherapy with rh-endostatin plus NP or NP alone. The numbers of activated circulating endothelial cells (aCECs) were measured by flow cytometry. Chest computed tomography was performed to evaluate the efficacy after 2 cycles of chemotherapy.</p><p><b>RESULTS</b>Cavitation occurred in 5 of 29 patients in the rh-endostatin plus NP group, but not in any case of the NP group. Of the 5 patients, there were 2 males and 3 females, with pathological types of 3 adenocarcinomas, 1 adenosquamous cell carcinoma and 1 sarcomatoid carcinoma. All of these 5 cases had a peripherally located tumor in the CT scan. There was only one cavity in each case and most of these were roundish. Four cavities were situated in the center of the tumor and another one was eccentric. There were 3 cavities with thin wall and 2 with thick wall. Their average diameter was 2.7 cm. No hemoptysis occurred in these 5 patients. The blood-supply of the tumors showed by perfusion CT images was inhibited in 3 cases after treatment. The average number of aCECs decreased from 323.2/10(5) to 33.0/10(5) after treatment.</p><p><b>CONCLUSION</b>Intratumoral cavitation is a peculiar imaging characteristics after anti-angiogenic therapy, which may be caused by inhibition of blood-supply to the tumor. CT perfusion imaging and measurement of activated circulating endothelial cells may be helpful to predict the efficacy of anti-angiogenic therapy combined with chemotherapy.</p>

Effects of platelet-derived membrane microparticles on angiogenesis in chick chorioallantoic membranes / 中国实验血液学杂志

This study was purposed to investigate the angiogenesis effect of platelet-derived membrane microparticles (PMPs) in chick chorioallantoic membranes (CAM). Thrombin were adopted to activate the platelets and then PMPs were obtained. PMPs were isolated by high speed centrifugation. Flow cytometry (FCM) was adopted to evaluate the efficiency of thrombin to produce PMPs and BCA method was adopted to evaluate the content of PMPs. PMPs were put into CAM and the effects of PMPs on angiogenesis in CAM were observed. The results indicated that after incubation for 72 hours at the concentration of 80 microg/ml PMPs, the vessel nets in a 'spoked-wheel' pattern were shown around mixed fibrous filter membranes, number of vessel ramification was 112.5 +/- 11.31 and ratio of vessel area/CAM area was 6.19 +/- 1.29%, but there were not localized allantoic vessels developing in the control group, the number of vessel ramification and ratio of vessel area/CAM area in control group were 82.6 +/- 8.05 and 1.78 +/- 0.33 respectively, so there was significant difference between PMP and control groups. In above mentioned conditions, the number of vessel ramification and ratio of vessel area/CAM area in VEGF group were 128.4 +/- 10.02 and 7.44 +/- 1.36 respectively, so there was no difference between PMP and VEGF groups. It is concluded that PMPs show promotive effect on the formation of capillaries in chick chorioallantoic membranes.

Test of activated plasma clotting time to assess efficacy of platelet transfusion / 中国实验血液学杂志

The study was aimed to investigate the value of activated plasma clotting time (APCT) for estimating the efficacy of platelet transfusion therapy. There were twenty patients with hematological diseases, who received transfusion of platelet, involved in the test. APCT was determined before and after transfusion of these patients, then APCT was contrasted with corresponding CCI and PPR. The results showed that 1 hour and 24 hour APCTs were shortened obviously. APCT before transfusion was (103.7 +/- 11.3) seconds, but the 1 hour and 24 hour APCTs were shortened to (60.0 +/- 9.7) seconds and (68.5 +/- 9.8) seconds respectively (P < 0.01). According to the judging criteria of CCI and PPR (CCI and PPR values at 1 and 24 hours after transfusion are < 7500, < 5000 and < 30%, < 20% respectively, the transfusion is invalid), two patients received invalid transfusion. Their 1 and 24 hour CCIs were 7415, 2966 and 6913, 4988 respectively. Their 1 and 24 hour PPRs were 28.0%, 11.2% and 25.2%, 14.1% respectively. One patient's PPR reached the standard of invalid transfusion, but his CCI showed a valid transfusion he received. Two patients' PPR reached the standard of invalid transfusion, but their 1 hour CCI reached the standard of valid transfusion, and their 24 hour CCI reached the standard of invalid transfusion. It is concluded that APCT reflects the variations of quantity and quality of platelet simultaneously, and can evaluate precisely the efficacy of platelet transfusion.

Cold storage of rabbit platelet suspension by adding uridine diphosphate galactose / 中国实验血液学杂志

This study was aimed to investigate the method to cold-store platelets with uridine diphosphate galactose (UDP-Gal). Rabbit heart blood was prepared for concentrated platelet suspension to which UDP-Gal was added, and then stored for ten days in 4 degrees C refrigerator. Thereafter, platelet count, mean platelet volume (MPV), platelet distributing width (PDW), platelet aggregation function, platelet activity to urge coagulation including PF3aT and APCT and apoptosis were determined. Meanwhile, survival time in vivo was tested after cold-stored rabbit platelets labeled with Cr51 were transfused into rabbits. The results showed that there was not significant difference for Plt count, MPV, PDW, PF3aT and APCT between UDP-Gal cold-stored platelet group and fresh platelet group (P > 0.05). On the contrary, platelet count decreased significantly, MPV, PDW jumped and PF3aT and APCT went down in cold control group as compared with fresh platelet group (P < 0.01). Apoptosis increased in UDP-Gal cold-stored platelet group as compared with fresh platelet group (P < 0.05), but was significantly lower than that in cold control group (P < 0.01). Although PagT (inducing reagent: C-PG) decreased, it could still be above 50% of fresh platelet. Survival time in rabbit in vivo was close between UDP-Gal cold-stored platelet group and fresh platelet group (P < 0.05). Survival rate in seventy-two hours after transfusion in the fresh platelet group, UDP-Gal cold-stored platelet group and cold control group was 57.5% +/- 7.2%, 50.3% +/- 6.3% and 0.1% +/- 0.5% respectively. It is concluded that the UDP-Gal can well protect cold-stored rabbit platelets and prolong the survival time of cold-stored platelets in vivo.

Influence of raising oxygen content on function of platelet concentrate during preservation / 中国实验血液学杂志

To explore the influence of raising oxygen (dissolved oxygen) content on function of platelet concentrate, the platelet concentrate was prepared by a CS-3000 plus blood cell separator. Experiments were divided into 2 groups: test group and control group. After raising oxygen content in platelet plasma under sterile operation, the platelet samples of two groups were preserved in oscillator with horizontal oscillation at 22 +/- 2 degrees C. The platelet count, platelet aggregation rate, lactic acid content and CD62p expression level of platelet were detected on 0, 1, 2, 3, 4, 5 days of platelet preservation. The results showed that the platelet count and platelet aggregation rate decreased with prolongation of preserved time, while the lactic acid content and CD62p expression level of platelet increased gradually. Compared with control group, there were significant differences in aggregation rate of platelet preserved for 2-3 days, and in CD62p expression level of platelet preserved for 1-3 days, while significant difference was found in lactic acid content of platelet preserved for 1-3 days. It is concluded that raising content of oxygen in platelet plasma can provide more oxygen to compensate oxygen supply deficiency for platelet metabolism and improve the efficiency of platelet oxygenic metabolism and the quality of platelet during preservation.

Value of imaging in the diagnosis of primary malignant fibrous histiocytoma of bone / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the imaging feature of primary malignant fibrous histiocytoma of the bone (PBMFH) by the conventional radiography, CT and MRI, and to evaluate these different imaging methods in its diagnosis.</p><p><b>METHODS</b>The imaging data of conventional radiography, CT and MRI of 35 patients with pathologically confirmed PBMFH were retrospectively analyzed.</p><p><b>RESULTS</b>Though the imaging appearance of PBMFH varied in different cases, all the imaging findings of malignant bone tumors were revealed. The common imaging appearance on the conventional radiography and CT were eccentric, aggressive, osteolytic destructions of various types located at the ends of extremities with extraosseous soft tissue masses, but periosteal reaction was rare. Heterogeneous signal intensities on T(1)WI and T(2)WI were common MRI changes but not specific.</p><p><b>CONCLUSION</b>Primary malignant bone fibrous histiocytoma, a rare primary malignant bone tumor, is most frequently located in the long bone. Conventional radiography is still the first and main choice and is taken as an essential means of diagnosis. CT and MRI are quite important in demonstrating the details and extent of the disease such as soft tissue, cortical destruction, periosteal reaction, calcification and necrosis. The imaging characteristics may be of value in differentiating MFH from the other malignant bone tumors. Furthermore, MRI may also be valuable in assessing the efficacy of chemotherapy and/or radiation therapy, as well as in distinguishing recurrence from postoperative or post-radiation changes.</p>

Application of cationic propyl gallate as inducer of thrombocyte aggregation for evaluating the platelet function of platelet donors / 中国实验血液学杂志

The purpose of study was to investigate the feasibility of the application of cationic propyl gallate (C-PG) as inducer of platelet aggregation for evaluating the platelet function of single-donor plateletpheresis and identifying the incidence of defective platelet function among donors. Experiments were as follows: 3 healthy volunteers' platelet aggregation induced by 100-300 micromol/L C-PG was determined by LG-PABER analyzer to observe the effect of C-PG concentration on platelet aggregation; 30 healthy volunteers' platelet aggregation before and 24 hours after administration of 200-400 mg acetylsalicylic acid (ASA) was examined after induction by 200 micromol/L C-PG for determining the cut-off value to discriminate platelet dysfunction donors; the platelet aggregation of 483 platelet donors was detected and the activated plasma clotting time (APCT) of donors who have deficiency in platelet aggregation was examined for investigating the incidence of defective platelet function among donors. The results showed that platelets were activated by C-PG induction in a dose dependent manner, when concentration of C-PG reached 200 micromol/L, the percentage of platelet aggregation was highest. It significantly decreased after 24 hours with ASA than that before the administration (P < 0.001), especially in 180 seconds induced by C-PG. If cut-off point was fixed on the platelet aggregation < 20% in 180 seconds, donors of platelet dysfunction can be selected effectively. 25 of defective platelet aggregation function among 483 donors were detected, and 11 out of 25 platelet dysfunction donors had the deficiency in procoagulant activity with prolonged APCT. It is concluded that C-PG as inducer of platelet aggregation is feasible to screen the platelet function of donors. Five percent of platelet donors has function defect examined by C-PG as inducer of platelet aggregation.