Exploration of the relationship between the storage time of leukodepleted red blood cell and transfusion adverse reactions / 中国输血杂志

【Objective】 To explore the relationship between the storage time of leukodepleted red blood cells and transfusion adverse reactions by analyzing the occurrence of transfusion adverse reactions of patients after leukodepleted red blood cells transfusion from four hospitals. 【Methods】 By using the electronic medical record management system, the collection and transfusion dates of leukodepleted red blood cells from four hospitals in Enshi Prefecture from 2018 to 2022, as well as the information on transfusion adverse reactions, were retrieved. 【Results】 From 2018 to 2022, a total of 697 61 bags of leukodepleted red blood cells were transfused in four hospitals, resulting in 166 cases of transfusion adverse reactions, among which 93 were allergic reactions, 63 were non hemolytic febrile reactions, and 10 were others, with a total incidence rate of transfusion adverse reactions at 0.24%. The average storage time of leukodepleted red blood cells with and without transfusion adverse reactions was (20.25±6.31) and (19.88±5.50) days, respectively. With a storage time of 7 days as the threshold, the incidence of transfusion adverse reactions was the lowest for a storage time of 15~21 days. The incidence of transfusion adverse reactions of leukodepleted red blood cells in two groups (with storage days ≤21 days and >21 days) was not statistically significant(P>0.05). 【Conclusion】 Allergic reactions were the main type of transfusion adverse reaction caused by leukodepleted red blood cells, and the incidence of transfusion adverse reactions decreased and then increased with the prolongation of the storage time of leukodepleted red blood cells. There was no significant difference in the incidence of transfusion adverse reactions with leukodepleted red blood cells stored for ≤ 21 days and >21 days.

Clinical features and prognosis of patients hospitalized with heart failure and low T3 syndrome / 中华内科杂志

Objective: To investigate the association between triiodothyronine (T3) and inflammatory factors, and its potential effect on long-term outcomes in hospitalized patients with heart failure (HF). Methods: A total of 2 475 patients with HF admitted in Heart Failure Care Unit were consecutively enrolled in this retrospective cohort study from December 2006 to June 2018. Patients were divided into low T3 syndrome group (n=610, 24.6%) and normal thyroid function group (n=1 865, 75.4%). The median follow-up time was 2.9 (1.0, 5.0) years. A total of 1 048 all-cause deaths were recorded at the final follow-up. The effects of free T3 (FT3) and high-sensitivity C-reactive protein (hsCRP) on the risk of all-cause death were evaluated by Cox regression analysis and Kaplan-Meier analysis. Results: The age of the total population was 19-95 (57±16) years, 1 823 cases (73.7%) were male. Compared to those with normal thyroid function, albumin [(36.5±5.4) vs. (40.7±4.7) g/L], hemoglobin [(129.4±25.1) vs. (140.6±20.6) g/L], total cholesterol [3.6 (3.0, 4.4) vs. 4.2 (3.5, 4.9) mmol/L] (all P<0.001) were lower, Whereas age [(60.5±16.0) vs. (55.2±15.4) years], creatinine [105.0 (83.6, 137.0) vs. 87.8 (75.6, 106.3) mmol/L], log N-terminal B-type natriuretic peptide [(8.2±1.3) vs. (7.2±1.4) ng/L] were higher in LT3S patients (all P<0.001). In Kaplan-Meier survival analysis, patients with lower FT3 and higher hsCRP had significantly lower cumulative survival (P<0.001), lower FT3 combined with higher hsCRP subgroup had the highest risk of all-cause death (Ptrend<0.001). In multivariate Cox regression analysis, LT3S was an independent predictor of all-cause mortality (HR=1.40, 95%CI 1.16-1.69, P<0.001). Conclusion: LT3S is an independent predictor of poor prognosis in patients with heart failure. FT3 combined with hsCRP improve the predictive value of all-cause death in hospitalized patients with heart failure.

Risk factors and prognostic value of heart-type fatty acid binding protein in patients with heart failure / 中华检验医学杂志

Objective:To investigate the prognostic value and related factors of heart-type fatty acid binding protein (H-FABP) in patients with heart failure.Methods:A total of 877 consecutive patients who were admitted to heart failure care unit of Fuwai hospital and diagnosed as heart failure from July 2015 to July 2017 were enrolled in this study. Baseline serum H-FABP concentration was measured by fluorescence lateral flow immunoassay. According to serum H-FABP levels, patients were divided into three groups: low H-FABP group (H-FABP≤4.04 ng/ml, n=292), middle H-FABP group (H-FABP 4.04-7.02 ng/ml, n=292) and high H-FABP group (H-FABP≥7.02 ng/ml, n=293). The general clinical characteristics were collected and compared among the three groups. According to whether heart failure was caused by coronary artery disease or not, patients with heart failure were divided into ischemic heart failure and non-ischemic heart failure. Multivariate linear regression analysis was performed to explore the independent risk factors of H-FABP. The primary endpoint events were the composite of all-cause death or heart transplantation. Multivariate Cox regression analyses, receiver operating characteristic (ROC) curves, risk prediction tests with multivariate Cox regression model and Kaplan-Meier analyses were conducted to investigate the relationship between H-FABP and the prognosis of heart failure. Results:Multivariate linear regression analysis showed that age, coronary artery disease, alanine aminotransferase, uric acid and N-terminal pro-B type natriuretic peptide (NT-proBNP) were positively associated with H-FABP (β=0.012, 0.238, 0.001, 0.345 and 0.063 respectively,all P<0.05), while female, hemoglobin, albumin, sodium, and estimated glomerular filtration rate (eGFR) were negatively associated with H-FABP (β=-0.184, -0.006, -0.016, -0.034 and -0.006 respectively, all P<0.05). One hundred and nineteen patients (13.6%) lost to follow-up, and 246 patients (32.5%) suffered from all-cause death or heart transplantation during the median follow-up duration of 931 (412-1 185) days. Multivariate Cox regression analysis showed that baseline H-FABP (log 2H-FABP) level was the independent predictor of all-cause death or heart transplantation in patients with heart failure ( HR=1.39, P<0.001). ROC curves showed that baseline H-FABP was a predictor of all-cause death or heart transplantation in patients with heart failure within 3 months, 1 year and 2 years (areas under the curves were 0.69, 0.69 and 0.71 respectively), and the best cut-off values were 5.85 ng/ml, 6.54 ng/ml and 6.54 ng/ml respectively. Risk prediction test with multivariate Cox regression model showed that baseline H-FABP could provide additional prognostic value in predicting all-cause death or heart transplantation for patients with heart failure on top of basic model and baseline NT-proBNP ( P<0.001). Taking 6.54 ng/ml and trisected levels of H-FABP as cut-off values respectively, Kaplan-Meier analyses showed that the survival rates were significantly different among the two or three groups ( P<0.001). Subgroup analyses showed that baseline H-FABP (log 2H-FABP) level was an independent predictor of all-cause death or heart transplantation in patients with ischemic heart failure ( HR=1.74, P<0.001), as well as in patients with non-ischemic heart failure ( HR=1.28, P=0.027). Conclusions:Age, sex, coronary artery disease, hemoglobin, albumin, alanine aminotransferase, sodium, eGFR, uric acid and NT-proBNP are associated with H-FABP level. Baseline H-FABP level is an independent predictor of all-cause death or heart transplantation in patients with heart failure. On top of basic model and baseline NT-proBNP, baseline H-FABP could provide additional prognostic value in predicting adverse events for patients with heart failure.

Detection of SMO gene mutations in odontogenic keratocyst / 中华口腔医学杂志

Objective: To detect the SMO mutations in odontogenic keratocyst (OKC) and to explore the mechanism behind. Methods: Patients with OKC who received treatment in the Department of Oral and Maxillofacial Surgery, School and Hospital of Stomatology,Peking University, from September 2012 to June 2017 were enrolled. OKC samples from 10 patients diagnosed as naevoid basal cell carcinoma syndrome (NBCCS)-related OKC (4 females and 6 males) and 20 patients diagnosed as sporadic OKC (7 females and 13 males) were collected. Genomic DNAs were extracted from fibrous capsules and epithelial lining respectively. SMO mutations were detected and analyzed by Sanger sequencing. Results: Three SMO mutations were found in one NBCCS-associated OKC who carrying c.2081C>G (p.P694R) mutation) and two sporadic OKC who carrying c.907C>T (p.L303F) mutation and c.1247_1248delinsAA (p.G416E), respectively), among which the first two mutations were novel mutations that had not been reported before. Besides, two mutations in sporadic OKC were not paired with PTCH1 mutations. Conclusions: In addition to PTCH1 gene mutations, SMO gene mutations also exist in OKC which might be related to the development of OKC.

Clinical characteristics of hypertrophic cardiomyopathy and restricted cardiomyopathy patients complicating with intracardiac thrombosis / 中华心血管病杂志

Objective: To investigate the clinical characteristics of patients with hypertrophic cardiomyopathy (HCM) and restrictive cardiomyopathy (RCM) complicating with intracardiac thrombosis. Methods: This is a retrospective observational study. Consecutive patients diagnosed with HCM or RCM and complicated with intracardiac thrombosis (including left and right atrium or ventricular thrombosis), who were admitted to the Heart Failure Care Unit of Fuwai Hospital, Chinese Academy of Medical Sciences, from September 2008 to September 2018, were enrolled in this study. Patients with myocardial infarction were excluded. The general clinical data of the enrolled patients, including demographic data, major complications, laboratory indicators, echocardiographic indicators, drug application and distribution of intracardiac thrombosis, were collected from electronic medical record system and analyzed. Results: A total of 98 patients were enrolled in this study, including 52 patients (53.1%) with HCM and 46 patients (46.9%) with RCM. The most common comorbidity was atrial fibrillation/flutter: 40 patients (76.9%) in HCM group and 36 patients (78.3%) in RCM group. Majority of patients received oral anticoagulants treatment: 43 patients (82.7%) in HCM group and 35 patients (76.1%) in RCM group. Intracardiac thrombosis was mainly located in the left atrium in both HCM group (39 cases (75.0%)) and RCM group (32 cases (69.6%)). Thrombosis was found in ≥ 2 chambers in 7 patients (7.1%). Rate of left atrial thrombosis was the highest (81.6% (62/76)) in HCM and RCM patients complicating with atrial fibrillation/flutter. Intra-aneurysmal thrombosis occurred in 4 out of 5 patients complicated with apical left ventricular aneurysm. The rate of left ventricular thrombosis in patients with left ventricular ejection fraction≥50% was 7.4% (4/54), which was significantly lower than that in patients with left ventricular ejection fraction<50% (34.5%(10/29)) (P<0.01). Conclusion: There are certain distribution characteristics of HCM and RCM patients with intracardiac thrombosis, and the left atrium is the most common site of thrombosis, more attention should be paid in HCM and RCM patients on the diagnosis and treatment of intracardiac thrombosis.

Associations between physical activity, screen time and anxiety, sleep quality among Chinese college students / 中国学校卫生

Objective@#To explore associations between physical activity, screen time and anxiety, sleep quality among college students in Shanghai, and to provide a reference for relevant prevention and control.@*Methods@#By using cluster random sampling method, a total of 4 964 students from grade 1 to grade 2 in 3 universities from 3 districts of Shanghai were enrolled. Self-Rating Anxiety Scale, Pittsburgh Sleep Quality Index and International Physical Activity Questionnaire were used to assess the level of anxiety, sleep quality and physical activity.@*Results@#The reporting rate of anxiety symptoms among students was 9.7%(8.7% for males and 11.4% for females) and the prevalence of poor sleep quality was 55.0%(51.8% for males and 60.4% for females), there was significant gender differences in anxiety symptoms and poor sleep quality rate(χ2=9.92, 34.81, P<0.01). Among male students, with adjustment of age, BMI and lifestyle, those who met neither physical activity nor screen time recommendations had 2.23(95%CI=1.31-3.79) and 1.48(95%CI=1.13-1.94) times risks for anxiety and poor sleep quality than those meeting both recommendations. Among girls, there was a significant association between screen time and anxiety(aOR=1.61, 95%CI=1.18-2.21). However, physical activity was not associated with anxiety and sleep quality.@*Conclusion@#High screen time and physical inactivity may increase the risk of anxiety and poor sleep quality among male college students, and screen time may also increase the risk of anxiety among female college students.

Clinical Feature Analysis for Heart Failure in 32 Hypertrophic Cardiomyopathy Patients Presented as Restrictive Cardiomyopathy / 中国循环杂志

Objective: To analyze the clinical features for heart failure (HF) in hypertrophic cardiomyopathy patients presented as restrictive cardiomyopathy. Methods: We retrospectively studied 32 hypertrophic cardiomyopathy combining HF patients with NYHA grade III-IV presented as restrictive cardiomyopathy and summarized their clinical features with the outcomes of in-hospital management. Results: Echocardiography found restrictive cardiomyopathy changes in all 32 severe hypertrophic cardiomyopathy combining HF patients as both atriums were enlarged and the size of left ventricle was normal; 84.4％ patients with normal LVEF (>50％) and 15.6％ with LVEF<50％; 37.5％ patients with enlarged right ventricle. HF history was from 10 days to 35 years at the mean of 8.3 years. 75％ patients appeared whole heart failure, the main symptoms were dyspnea, edema, some patients had syncope and angina. There were 8 patients with respiratory failure, 2 with cardiac shock, 13 with medium to large amount of pleural effusion and ascites; 90％ patients combining paroxysmal or persistentatrial fibrillation (AF), 8 patients received pacemaker implantation due to slow tachycardia. The in-hospital ventricular tachycardia or ventricular fibrillation occurred in 3 patients, 2 of them were successfully rescued by electrical cardio-version and received implantable cardioverter defibrillator(ICD), 1 died for failed cardio-pulmonary resuscitation; 6 patients had heart transplantation.Conclusion: Severe hypertrophic cardiomyopathy combining HF patients presented as restrictive cardiomyopathy were usually at the late stage in critical condition with various complications even they could have normal size of left ventricle and LVEF, some patients may need heart transplantation.

Nrf2-NF-κB pathway axes, epigenetic regulation and traditional Chinese medicine (natural medicine) to treat type 2 diabetes / 中国中药杂志

Diabetes mellitus is a characterized by high blood sugar metabolic disease, is a lifelong disease with a high incidence of major hazards. Prevention and treatment of diabetes and its complications has become a serious challenge and arduous task facing the world pharmaceutical researchers. Oxidative stress, Nfr2-NF-κB signaling axis related epigenomic genes have the apparent close relationship with type 2 diabetes,those have become one of the key focus and effective way to explore its pathogenesis, mechanism and drug screening. This paper systematically summarizes the current stage research regulating the key proteins, mRNA about Nrf2-NF-κB axis pathway and epigenomics for treatment of type 2 diabetes and the mechanism of traditional Chinese medicine and natural medicine (component) in the treatment of type 2 diabetes, hoping to provide some innovative research ideas for finding new drugs of the treatment of diabetes from traditional Chinese medicine and natural medicine.

Application of Child-Turcotte-Pugh Scores in Predicting the Risk of Death for In-hospital Heart Failure Patients / 中国循环杂志

Objective: Heart failure (HF) patients are usually associated with liver function impairment, Child-Turcotte-Pugh (CTP) scores can evaluate liver function, but its effect in HF patients has been unclear. We want to study the application of CTP scores in predicting the risk of death for in-hospital HF patients. Methods: A total of 1180 consecutive in-hospital HF patients were enrolled. According to CTP scores evaluated liver function at admission, the patients were divided into 3 groups: CTP grade A group, n=951, CTP grade B group, n=206 and CTP grade C group, n=23. The endpoint of this study was all-cause death. Results: There were 180 patients died at 1 year follow-up period, the in-hospital and 1 year mortalities were increased with the elevated CTP grades accordingly: for in-hospital mortalities in CTP grade A, B and C groups were (0.8%, 11.7% and 56.5%) respectively, P Conclusion: CTP scores may independently predict the risk of death for in-hospital HF patients, the levels of CTP scores might be used for evaluating the efficacy of in-hospital treatment.

Metformin Influences the Effect of Adipocytes on Leukemia Cell Chemoresistance through Regulation of Adipogenesis / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the effect of metformin on 3T3-L1 preadipocyte's differentiation and consequently observe the anti-proliferative effects of metformin-treated adipocytes on leukemia cells.</p><p><b>METHODS</b>Different concentrations of metformin were added in 3T3-L1 preadipocytes to induce maturation, the matured adipocytes were detected by oil red O staining and quantified by absorbance value (OD). Real-time PCR was used to detect the mRNA expression level of the key adipogenic genes PPARγ, C/EBPα and FABP4(ap2). The adipocytes were co-cultured with GFP+-THP-1 cells, 1 µg/ml of cytarabine(Ara-C) was added and incubated for 48 hours, the flow cytometry was used to detect the apoptosis rate of GFP+-THP-1 cells. Adipocyte supernatant was collected and mixed with equal volume of tumor lysat medium (RPMI 1640) at 1:1 to culture tumor cells. The leukemia cell proliferation activity was detected by CCK-8; after 48 hours of adding 1 µg/ml Ara-C, the protective effect on chemotherapy was assayed by using cytometer.</p><p><b>RESULTS</b>The metformin lowered the OD value, and the expression levels of both adipogenic genes C/EBPα and FABP4 were lower than those of controls, while the expression level of PPARγ mRNA was not significantly changed, the apoptosis rate of leukemia cells co-caltured with metformin-treated adipocytes was higher than that of co-cultured cells without metformin treatment. The adipocytes promoted the leukimia cell proliferation and protected leukemia cells from chemotherapy, which could be abrogated by metformin.</p><p><b>CONCLUSION</b>The metformin can inhibit the differentiation of 3T3-L1 preadipocytes into adipocytes, and can regulate the protective effect of adipocytes on the apoptosis, proliferation and chemotherapy of leukemia cells.</p>

Comparison of Adipogenesis and Adipocyte Functions of 3T3-L1 Cells and Human Bone Marrow Mesenchymal Stem Cells In Vitro / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To compare the adipogenesis and the adipocyte function between 3T3-L1 cells and human bone marrow mesenchymal stem cells (MSCs) in vitro.</p><p><b>METHODS</b>By density gradient centrifugation and adherent culture, the MSCs were isolated from human bone marrow and purified. The cell morphology was observed under an inverted microscope. After the induction of adipogenic differentiation, the differentiation level was detected by oil red O staining and OD values. The expression of PPARγ, FABP4 and C/EBPα mRNA was detected by quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR). Adipocytes and THP-1 cells were co-cultured by adding 1 µg/ml cytarabine. The ability of chemotherapy resistance was measured after 48 h.</p><p><b>RESULTS</b>The Oil Red O staining and measuring the absorbance showed that the lipid content in 3T3-L1 cells group was more than that in MSCs group, and the OD value was higher than that in MSCs group (P < 0.05). The results of qRT-PCR showed that the relative expression of PPARγ, FABP4 and C/EBPα mRNA of 3T3-L1-derived adipocytes was higher than that of human bone marrow MSCs-derived adipocytes (P < 0.05). Coculture experiments showed that the number of viable THP-1 cells in the group containing adipocytes was more than that in the control group (P < 0.05). The difference between 3T3-L1 cell group and MSC group was statistically significant (P < 0.05).</p><p><b>CONCLUSION</b>The ability of adipogenesis of 3T3-L1 cells is higher than that of human bone marrow MSCs in vitro. Adipocytes can protect THP-1 cell line against cytarabine, and the effect of adipocytes from 3T3-L1 cell group is greater than that from the human bone marrow MSC group.</p>

Research advances on abnormal marrow fibre in leukemia / 中国实验血液学杂志

The genesis and development of leukemia not only associate to intrinsic factors, but also relate with the fibrous hyperplasia in the bone marrow. This review mainly focuses on the interaction between fiber-producing cells and leukemia cells, the relationship between fibrous hyperplasia and prognosis of leukemia, the regulation of TGF-beta, PDGF and other cytokines, the underlying mechanism of fibrous hyperplasia so as to explore the potential therapeutic targets for improving the prognosis of leukemia.

Correlation between ALIP-like cluster and fibrous proliferation in bone marrow of patients with acute myeloid leukemia in CR phase / 中国实验血液学杂志

This study was purposed to elucidate the prognostic values of ALIP (abnormal location of immature precursors)-like clusters and fibrous proliferation in bone marrow of AML patients in CR phase and the correlation between them. The bone marrow biopsy sections from 47 AML patients during admitting to relapse or till lost follow-up were examined retrospectively. The 47 patients were divided into pre-relapsed group and non-relapsed group according to relapse or not at end of follow-up. The concentration of ALIP-like cluster and reticulin fiber density (RFD) in sections were compared between the two groups respectively, the prognostic value of these two factors and the underlying relationship between them were estimated by statistical analysis. The results showed that ALIP-like cluster was (3.46 ± 2.71)/mm(2) during CR and RFD was (2.76% ± 1.50%) in pre-relapsed cases, which both were higher than those in non-relapsed cases (P < 0.05). Cases with ALIP-like cluster over 4/mm(2) or with RFD>1.68% showed high relapse rates of 89.5% or 95.2% respectively. RFD were (2.47% ± 2.48%) and (2.44% ± 2.23%) in cases with >4/mm(2) or ≤ 4/mm(2) respectively, there was no statistical significance (P > 0.05) . Meanwhile, the amount of ALIP-like cluster in CR not related with the paired RFD (r = 0.057, P > 0.05). It is concluded that both ALIP-like cluster in CR and RFD are poor prognostic factors for heralding early relapse. However, ALIP-like cluster and RFD show no correlation, and suggest that forming of ALIP not depends on fibrogenesis.

Transient dynamic analysis and optimization of a new customized lingual self-locking appliance during occluding / 医用生物力学

Objective To study the characteristics of stress distributions on a customized lingual self-locking orthodontic appliance under transient occlusal force and optimize its structure. Methods A whole 3D model including denture, appliances and wire was established by CT scanning, reverse engineering method and CAD technology; transient nonlinear dynamic analysis on this model during occluding and its structural optimization were conducted, and the optimized lingual appliance was made based on rapid prototyping technology to verify reliability of the finite element model. Results The equivalent stress on the bracket bottom was larger than that on other parts of the bracket; the maximum equivalent stress on the bracket cover was decreased by 60.9% after installing a reinforcing rib on it, which could effectively prevent stress concentration caused by the contact between the arch wire and bracket cover. The simulation results fundamentally agreed with the loading experiment on the bracket cover. Conclusions For lingual orthodontic treatment in clinic, the relative position between interaction points of the occlusal force and brackets should be concerned so as to avoid impairing the self-locking function; through optimizing the appliance design, the elastic potential energy of arch wire can be transferred more effectively to the teeth and reduce losses of the orthodontic force.

Influence of pre-ALIP number and its distance from trabeculae on AML relapse / 中国实验血液学杂志

This study was purposed to detect the abnormal quantity and localization of pre-ALIP in bone marrow of acute myelocytic leukemia patients (AML) during the complete remission (CR) and investigate their correlation with AML relapse. The bone marrow biopsy and prognosis of 62 patients with CR were retrospectively analyzed. The bone marrow was divided into the pre-relapse group and the no-relapse group according to prognosis of patients. In order to clarify the correlation of abnormal quantity and localization of pre-ALIP with AML relapse, the number of single and double-cluster precursor cells and the sum of both were calculated, and their distance from bone trabeculae was surveyed with the computer image segment method. The results showed that the number of pre-ALIP in pre-relapse group (11 ± 11.71/mm(2)) and no-relapse group (8.33 ± 9.17/mm(2)) were more than that in normal control group (5.29 ± 4.00) (P < 0.01). The number of pre-ALIP more than 11/mm(2) was observed in 17 among all AML patients, and out of them 12 patients with pre-ALIP number >11/mm(2) (70.6) were found in the pre-relapse group, which was higher than that in no-relapse group (P < 0.05). While the distance between pre-ALIP and trabeculae [(341.31 ± 266.16) µm] in pre-relapse group showed the tendency of migrating to the intermediate zone of bone trabeculae, compared with that in no-relapse group [(242.41 ± 174.65) µm, P < 0.01]. Moreover, about 77.8 of 18 patients showed the distance of pre-ALIP from trabeculae was more than 341 µm in the pre-relapse group, and significantly higher than that in no-relapse group (P < 0.01). It is concluded that the average number of "pre-ALIP" more than 11/mm(2) or the average distance from trabeculae longer than 341 µm in bone marrow sections during CR may be the indicators for early relapse of AML.

Results of requesting a second consecutive sperm ejaculate on the day of oocyte pick-up in assisted reproductive technology / 中华男科学杂志

<p><b>OBJECTIVE</b>To evaluate the results of requesting a second consecutive sperm ejaculate in order to reduce ICSI cycles by PESA or TESE on the day of oocyte pick-up in assisted reproductive technology (ART).</p><p><b>METHODS</b>We collected 68 semen samples as a second consecutive ejaculate from 34 men, compared the semen volume and sperm concentration, motility and total count between the first and the second ejaculation, and analyzed the laboratory results and clinical outcomes of fertilization with the mixed sperm.</p><p><b>RESULTS</b>The 34 males ejaculated twice within 4 hours by masturbation, with an interval of 26-183 (94.9 +/- 39.8) minutes between the first and second ejaculation. The volume of the first ejaculate was (2.0 +/- 1.4) ml, significantly higher than that of the second ([1.5 +/- 0.9] ml) (P = 0.007), although the numbers of motile sperm and grade a + b sperm of the first ([40.8 +/- 25.3]% and [30.9 +/- 22.4]%) were significantly lower than those of the second ([52.2 +/- 21.1]% and [39.9 +/- 17.5]%) (P < 0.05). There were no statistically significant differences in the sperm concentration or total sperm count between the two ejaculates (P > 0.05). The ICSI, IVF + ICSI, and IVF cycles were 3, 3 and 28 respectively among the 34 couples undergoing ART. The number of retrieved oocytes, normal fertilization rate, high quality embryo rate and frozen cycles/fresh transfer cycles ratio were 15.5 +/- 8.7, 57.0% (247/433), 58.7% (145/247) and 20/24 for the IVF cycle, 21.7 +/- 8.3, 61.5% (40/65), 67.5% (27/40) and 3/2 for the ICSI cycle, and 10.0 +/- 2.6, 72.4% (21/29), 66.7% (14/21) and 3/3 for the IVF + ICSI cycle. Fourteen live births were achieved out of the 18 pregnancies, including 6 healthy boys and 9 healthy girls.</p><p><b>CONCLUSION</b>A clinical pregnancy rate of >30% can be achieved by requesting a second consecutive sperm ejaculate on the day of oocyte pick-up in order to collect more sperm and/or increase the total number of motile sperm for ART. And this method can avoid other invasive sperm processing techniques and the need of unnecessary micromanipulative fertilization.</p>