Objective To investigate the impact of human leukocyte antigen(HLA)functional epitope mismatch(EM)on the efficacy of platelet transfusion in patients with hematological diseases.Methods HLA genotyping was performed on platelet donors and patients with hematological diseases who applied for platelet serological cross-matching and HLA antigen matching from June 2021 to June 2023 by PCR-SBT method.HLA platelet matching was based on the principle of CREG to se-lect donors for patients.HLA Matchmaker 4.0 software was used to analyze donor-recipient HLA EM information.The expres-sion level and gene distribution of related HLA functional epitope(Eplet)were searched from the international HLA Epitope registry website(www.Epregistry.com.br).Retrospective analysis was conducted on clinical platelet transfusion data to evalu-ate the impact of HLA EM on platelet transfusion effectiveness.Results Platelet transfusion efficacy showed no correlation with gender and age,but it was associated with platelet matching strategy.When the total number of HLA EMs was less than 20,a lower total number of donor-recipient HLA EMs resulted in higher platelet transfusion efficiency(χ2=19.311,P=0.001)and higher average value of 24 h corrected count increment(CCI)(F=7.737,P<0.001).The total number of donor-recipient HLA EMs had negative correlation with actual 24 h CCI(Rho=-0.322,P<0.001).Further statistical analysis re-vealed that 17 Eplets were related to the effectiveness of platelet transfusion.The locus distribution of 17 Eplets might be u-nique to HLA-A(17.6%)or-B(64.7%)or shared between HLA-A and-B(17.6%),and its expression may be high(58.8%)or intermediate(41.2%).Conclusion The total number of donor-recipient HLA EMs is a crucial factor influencing platelet transfusion effectiveness,and several HLA Eplets associated with this effectiveness have been identified.

Objective: To investigate the breakthrough incidence of invasive fungal disease(IFD) and side effects of posaconazole as primary prophylaxis during induction chemotherapy for acute myeloid leukemia(AML). Methods: A total of 206 newly diagnosed AML patients admitted to our department during January 2016 and December 2018 were enrolled in the study. Exclusive criteria were as followings including patients diagnosed as acute promyelocytic leukemia; those who received intravenous antifungal therapy after admission or had history of IFD one month before induction chemotherapy, or those with functional insufficiency of vital organs and those older than 65. Forty-seven patients received posaconazole (posaconazole group), 61 cases received voriconazole (voriconazole group) and 98 cases did not receive any prophylaxis (control group) during induction chemotherapy. Prophylactic efficacy and safety between posaconazole and voriconazole were compared. Results: During induction chemotherapy, five possible cases of IFD occurred in posaconazole group (10.6%); while 11 cases (18.0%) were in voriconazole group including 7 possible, 3 probable and 1 proven. Thirty-five cases (35.7%) in control group were diagnosed as IFD including 19 possible, 11 probable and 5 proven ones. The incidences of IFD in posaconazole and voriconazole group were significantly lower than that in control group (P<0.05). The difference of posaconazole group and voriconazole group was not significant (P>0.05). The reported adverse events in posaconazole group were significantly lower than those in voriconazole group [12.8%(6/47) vs. 32.8%(20/61), P<0.05]. Conclusions Posaconazole and voriconazole decrease IFD as primary prophylaxis during induction chemotherapy in patients with AML. The prophylactic effect of IFD with posaconazole is similar as voriconazole, but posaconazole shows better safety.

Objective To analyze the prognostic impact of Ikaros family zinc finger 1(IKZF1)mutation on adult Philadelphia chromosome (Ph1) positive acute lymphoblastic leukemia (ALL) patients.Methods IKZF1 mutation was detected in 63 adult Phi positive ALL patients at diagnosis using capillary electrophoresis.Recruited patients were treated in our center and other three hospitals in Ningbo from January 2014 to January 2017.Clinical data were collected and retrospectively analyzed.Results Thirty-nine (61.9％) patients were positive IKZF1 mutation in this cohort.The white blood cell (WBC) count in IKZF1 mutation group was significantly higher than that of mutation negative group [(64.6±11.3)× 109/L vs.(33.7±5.6)×109/L,P＜0.05].Patients with WBC count over 30×109/L accounted for 56.4％ in IKZF1 mutation group.Complete remission (CR) rate in the IKZF1 mutation group was also lower than that of negative group after induction chemotherapy (64.1％ vs.75.0％,P＞0.05).IKZF1 was a negative prognostic factor but not independent factor for survival by univariate and multivariate analyses.Patients were divided into chemotherapy and allogeneic transplantation groups.The 3-year overall survival (OS) rate and 3-year leukemia-free survival (LFS) rate in IKZF1 mutation group were significantly lower than those of negative group in both transplantation group (42.3％ vs.59.3％;31.2％ vs.50.0％;respectively,both P＜0.05) and chemotherapy group (24.8％ vs.40.0％;19.0％ vs.34.3％;respectively,both P＜0.05).Conclusion IKZF1 mutation is a poor prognostic factor for adult Ph1 positive ALL patients.

Objective:To observe the effect of sciatic nerve pulsed radiofrequency (PRF) on the activation levels of microglia cells and astrocytes in spinal dorsal horn in the chronic constriction injury (CCI) rat models, and to explore the relationships between the analgesic mechanism of sciatic nerve PRF and the activation levels of microglia cells and astrocytes in the spinal dorsal horn.Methods:Forty male SD rats were randomly divided into CCI shamoperation+RPF sham group (SS group) , CCI sham-operation+RPF group (SP group) , CCI+RPF sham group (CS group) , CCI+RPF group (CP group) (n=10) .PRF was applied on the sciatic nerve on the 4th day after CCI operation for 120s, with a maximum temperature of 42℃.The mechanical withdrawal thresholds (MWT) and thermal paw withdrawal latencies (TWL) were measured to evaluate mechanical hyperalgesia and thermal hyperalgesia 1dbefore operation (D0) and 1, 3, 5, 7dafter operation (D1, D3, D5, D7) .Western blotting method was used to detect the protein expression levels of ionized calcium binding adapter molecule 1 (Iba-1) and glial fibrillary acidic protein (GFAP) in the ipsilateral spinal dorsal horn of L3-5after pain behavioral test (D7) .Results:Compared with SS group, the rats in CS group after CCI had the significant behavioral changes, such as hallux valgus, lameness, toe bending, and foot raising during walking;MWT and TWL were decreased significantly (P＜0.01) ;the expression levels of Iba-1and GFAP proteins in the ipsilateral spinal dorsal horn were increased significantly (P＜0.05) .Compared with CS group, the behavioral changes of the rats in CP group (D5, D7) such as hallux valgus, lameness, toe bending, and foot raising during walking were alleviated significantly;MWT and TWL were increased significantly (P＜0.01) ;the Iba-1protein expression level in spinal dorsal horn was downregulated significantly (P＜0.05) and the GFAP protein expression level did not change significantly (P＞0.05) .Conclusion:PRF on sciatic nerve can relieve the neuropathic pain (NP) of the CCI rat models;PRF on sciatic nerve can inhibit activation of the microglia cells in the spinal dorsal horn.The analgesic mechanism of PRF on sciatic nerve may be associated with the inhibition of the activation of microglia cells in the spinal dorsal horn.

Objective: To investigate the impact of FLT3-ITD and DNMT3A R882 double mutations to the prognosis of acute myeloid leukemia after allogeneic hematopoietic stem cell transplantation (allo-HSCT). Methods: FLT3-ITD, DNMT3A, C-kit, CEBPA, FLT3-TKD and NPM1 mutations were detected in 206 newly diagnosed AML patients by Sanger sequencing (M₃ and those received FLT3 inhibitor were excluded). Clinical data of AML patients were retrospectively analyzed to compare the prognosis of each gene mutation group. Results: ①Of 206 patients, 104 were male and 102 female with a median age of 38 (3-63) years, including 6 cases of M₀, 24 cases of M₁, 56 cases of M₂, 39 cases of M₄, 63 cases of M₅, 6 cases of M₆ and 12 unclassified cases. ②All 206 patients were divided into four groups according to the mutation gene at the time of diagnosis: FLT3-ITD⁺ DNMT3A R882⁺ group (group A), FLT3-ITD⁺ DNMT3A R882^- group (group B), FLT3-ITD^- DNMT3A R882⁺ group (group C) and FLT3-ITD^- DNMT3A R882^- groups (group D). Gender, leukocyte count at diagnosis, chromosome karyotype, the median age, FAB classification, disease status prior to transplantation, type of donor, conditioning regimen and GVHD were not significantly different between four groups (P>0.05). ③The 2-year cumulative recurrence rate (CIR) of group A was significantly higher than that of other groups [group A (72.2±2.6)%, group B (38.6±0.6)%, group C (36.8±1.6)%, group D (27.8±0.1)%, respectively, P<0.05], while the 2-year overall survival (OS) rate and 2-year leukocyte-free survival (LFS) rate were lower than those of other groups [group A (30.9±13.3)%, (11.3±10.2)%; group B (67.5±7.8)%, (47.9±8.4)%; group C (61.4±12.4)%, (56.8±12.5)%; group D (80.1±3.7)%, (79.7±3.6)%, respectively, P<0.05]. Conclusion AML patients with FLT3-ITD and DNMT3A R882 double mutations had a very high CIR and low OS, LFS after transplantation.

Objective To observe the clinical effect of sequential therapy with micafungin and reduced -dose voriconazole in prevention of invasive fungal infections in patients received allogeneic hematopoietic stem cell transplantion (Allo -HSCT).Methods 28 patients received the treatments for prevention of fungal infection with micafungin 50 mg per day from pretreatment to 30 days,then oral voriconazole at a dose of 1 00 mg two times per day until 90 days after Allo -HSCT.The occurrence of invasive fungal infection and the side effects of both medicine were observed during 1 80 days after Allo -HSCT.Results 8 patients(28.6%)developed above grade 2 acute graft verse host disease(GVHD),2 patients developed grade 3 GVHD among the 8 patients.Two case with GVHD were cured by voriconazole with the therapeutic dose who occurred probably pulmonary invasive fungal infection at two months after Allo -HSCT.There were no other patients diagnosed fungal infection.No toxic efect were observed during the clinical observation during treatment with micafungin.5 patients appeared mild liver function abnormalities during treatment with voriconazole,and liver dysfunction were improved by symptomatic treatment.2 cases developed transient auditory hallucination and visual impairment induced by voriconazole.Conclusion Micafungin and reduced -dose voricon-azole are effective and safe prophylaxis in prevention early invasive fungal infection after HSCT.

Objective To analyze the genetic quality of 24 domestic inbred strains mice using microsatellite loci panel.Methods Previously selected 30 microsatellite loci of mouse with high polymorphism and more allele numbers were used to synthesize corresponding fluorescently-labeled primers.Then the genomic DNA samples of each mouse were amplified by PCR and the products were analyzed by STR scanning to genotype the inbred strains of mice.Results Out of the 24 inbred strains, 15 inbred strains showed the same genotype within one strain at 30 loci.Among different strains, microsatellite loci indicated polymorphism which could be used to distinguish different strains.However, the rest 9 strains demonstrated polymorphism within strains.Conclusions Our stuoly provides a useful microsatellite panel to detect genetic quality of inbred mice and distinguish different strains with the optimized microsatellite loci.

Objective To explore the types of variations in the origin of left gastric artery (LGA) using MSCT angiography.Methods The abdominal MSCT angiography data of 1 500 patients were respectively reviewed,in thoses the abdominal aorta,celiac trunk,LGA,common hepatic artery (CHA),splenic artery (SA) and superior mesenteric artery (SMA)were shown clearly.The origins of the LGA and related artries were focused.A new typing method (types Ⅰ-Ⅹ) was established.And the incidence of various types was calculated.Results The normal anatomical origin (type Ⅰ) of LGAwas noted in 1 342 cases (1 342/1 500,89.47％).Eight types of LGA variant origin were identified in 70 cases (70/1 500,4.67％).LGA variant origin combined with celiomesenteric trunk (CMT) were observed in 47 cases (47/1 500,3.13％).The most common type of LGA origin variation was LGA originated from the abdominal aorta combined with CMT (type Ⅴ) which was found in 24 cases (24/1 500,1.60％).And the least common type was the namely LGA,SA,CHA and SMA arose independently from abdominal aorta (type Ⅵ) which was found in 3 cases (3/1 500,0.20 ％).LGA originated from SMA (type Ⅷ) was not found in all 1 500 cases.Conclusion There are many kinds of variations in the origin of LGA.The new typing method can contribute the comprehensive and intensive data for understanding the anatomical and radiographic features.

Objective To observe the expression of protein phosphates 1 nuclear targeting subunit (PNUTS) in the cochlea of D-galactose induced ageing mice. Methods Twenty Kunming mice, six weeks old, cleaning degree, were randomly divided into two groups, control group and D-galactose group, ten mice for each group. Mice in D-galactose group were administrated with D-galactose at a dose of 800 mg/(kg · d) by subcutaneous injection for eight weeks. Mice in control group were injected with the same volume of saline. After eight weeks, auditory brainstem responses (ABR) were collected to test the hearing thresholds of mice. Western blot assay was used to detect expressions of PNUTS and p53 protein. The expression and distribution of PNUTS in the cochlear Corti, spiral ganglion and striavascularis cells were observed by immunohistochemical (IHC) staining. Results There were no significant differences in ABRs at 8, 12 and 24 kHz between two groups. Protein expressions of PNUTS were located in the cochlear hair cells, spiral ganglion cells and striavascularis cells, and the expression level of cochlea was significantly decreased in D-galactose group than that in control group ( P<0.05). The expression level of p53 protein was significantly increased in D-galactose group than that in control group (P<0.01). Conclusion PNUTS is expressed in the normal mouse cochlea, and which is down-regulated in the cochlea of ageing mice induced by D-galactose.

AIM:To determine the biological feature of circulating endothelial cells (CECs) in acute promye-locytic leukemia ( APL) patients before and after treatment , and to analyze the relationship between CECs and the clinical characteristics .METHODS: The CECs were sorted from peripheral blood by magnetic-activated cell sorting and then counted by 3-color flow cytometry.The cells were identified by immunofluorescence staining for the expression of CD 146, CD31, CD144, VEGFR-2, CD45 and CD133.The CECs were cultured in vitro, and the tube formation and colony-forming rate were determined .RESULTS:Increased quantity of CECs was observed in CD 34 positive group and group with WBC >10 ×109/L (P<0.05).The quantity of CECs had a significant difference among low risk , medium risk and high risk groups (P<0.05).The positive rate of CD133 and quantity of CECs significantly reduced in 32 APL patients when they gain complete remission after treatment (P<0.05).The amount of tube formation and colony-forming rate were significant-ly reduced after treatment (P<0.05).The ratio of CECs quantity from APL patients after treatment to that before treatment had a negative correlation with arsenic concentration in urine on day 7 during As2O3 treatment (P<0.05).CONCLU-SION:Accurately counting CECs may be helpful for evaluating prognosis and designing treatment strategy .