Objective To compare the early embryonic developmental potential and clinical outcomes of oocytes matured in vivo and those matured by modified in vitro maturation(LVM)technology during the same controlled ovarian hyperstimulation(COH)cycle,and to explore the clinical application of melatonin-supplemented IVM technology.Methods 159 patients were recruited into the study.920 mature oocytes were collected during their COH cycles processed for conventional IVF/ICSI protocols,while 1 283 immature oocytes from the same cycles were matured in a melatonin-supplemented IVM medium before ICSI was performed.A retrospective analysis was conducted to compare the impact of conventional assisted reproductive technology and improved IVM technology on the outcomes of assisted reproductive therapy and pregnancy outcomes.Results Compared with mature oocytes collected from COH cycles treated with conventional IVF/ICSI,oocytes promoted by improved melatonin-supple-mented IVM technology had a lower rate of high-quality blastocyst formation.However,after embryo transfer,there was no significant difference in the clinical outcomes of mature oocytes obtained through two methods,including clinical pregnancy rate,full-term birth rate,neonatal length,and neonatal Apgar score.Conclusion The applica-tion of melatonin-supplemented IVM significantly increases the utilization of immature oocytes collected from COH cycles,improving the pregnancy outcomes of patients assisted by assisted reproductive technology.

We developed a pre-treatment method to remove interfering substances during quantification of 146S antigens in foot-and-mouth disease (FMD) vaccines by high performance size exclusion chromatography (HPSEC). Three methods, including ultracentrifugation, PEG precipitation and nuclease digestion, were optimized and compared for removal efficiency of the interfering impurities in FMD vaccines. Under optimized conditions, the 146S contents in two batches of FMD vaccines were determined to be 7.1 and 7.6 μg/mL by ultracentrifugation, 9.7 and 10.4 μg/mL by PEG precipitation, and 10.5 and 10.4 μg/mL by nuclease digestion. The optimal condition for nuclease digestion using Benzonase determined by response surface method was as follows: appending Benzonase into 200 μL of antigen phase to a final concentration of 421 U/mL and incubating at 25.1 °C for 1.29 h. This method has advantages including efficient removal of the interfering impurities, fast processing speed, and mild operating conditions. Then 12 bathes of FMD vaccines with different serotypes produced by 4 manufacturers were tested to verify the established treatment method. Results showed the method was applicable to various FMD vaccines with good reproducibility (RSD<5.3%, n=3). The developed method removed interference from impurities during quantification of 146S, and therefore would broaden the application of HPSEC in vaccine quality control and ensure the accuracy and reliability.
Animals ; Chromatography, Gel ; Foot-and-Mouth Disease ; Foot-and-Mouth Disease Virus ; Reproducibility of Results ; Viral Vaccines

Objective To evaluate the results of 2017 external quality assessment for newborn hemoglobinopathyand improve the quality of disease screening .Methods Each of 26 participating laboratories testing newborn hemoglobinopathy across the country received 5 batches of quality control blood spots ( Lot 201711-201715 ) in octorber 2017.Laboratories voluntarily participated in the survey and reported the results, methods, equipments and reagents information .Clinet EQA, and Microsoft Excel 2010 were used to perform statistical analysis on the laboratory test results .The rates of accuracy ( number of correct results/total number of submitted results ) were used for evaluating the performance of laboratories . Results 24 laboratories submitted the testing results with a return rate of 80.8%(21/26).The rates of accuracy for each lot were 100%(21/21), 90.5%(19/21), 90.5%(19/21), 57.1%(12/21) and 100%(21/21 ) respectively.Conclusions The results of this external quality assessment for newborn hemoglobinopathy is generally satisfactory , except for HbBarts′and HbA2.The screening laboratories should improve their quality control system , take timely measures to correct mistakes during the analytic period and improve the accuracy of screening tests for newborn hemoglobinopathy.

Objective To explore the protective effect of propofol on endothelial cells during heat stress and its protective effect to mitochondra. Methods Heat stress model of human umbilical vein endothelial cell was established when cells were incubated at 43℃ for 2h, then further incubted at 37℃, 5%CO2 for 6h. The experimental group was subdivided into six groups, including 37℃ group, 37℃ plus intralipid group (negative control group), 37℃ plus propofol group, 43℃ plus propofol group, 43℃ plus intralipid group, H2O2 plus propofol group (positive control group); Pretreated with 50μmol/L propofol, 0.2ml intralipid or 25μmol/L H2O2 before heat stress at 43℃, while the cells in the control group were incubated at 37℃. Cell viability was tested by CCK-8. ROS, mitochondrial membrane potential and the changes in mitochondrial permeability transition pore were determined by flow cytometry. The level of ATP was detected by fluorescein-luciferase. The changes of caspase-9 and caspase-3 were analyzed by Caspase Activity Assay Kit. Results HUVESs cell viability and damage of mitochondra were significantly decreased after heat stress. Compared with 43℃ heat stress group, pretreatment with propofol induced the recovery of cell viability and the ROS levels were significantly decreased in HUVEC cells (P<0.05). Meanwhile, the number of cells representing the decrease of mitochondrial membrane potential (the proportion of JC-1 monomer) was significantly decreased (P<0.05) by propofol. The average fluorescence intensity of calcein which representing the MPTP changes and intracellular ATP content was significantly increased (P<0.05). In addition, the activation of mitochondrial apoptotic pathway mediated by caspase-9/3 was also inhibited. Conclusions Propofol have anti-oxidative, anti-apoptosis and mitochondria protective effect against endothelial cell injury during heat stress.

Objective To observe the oxidative stress, integrity of lysosome and apoptosis of intestinal epithelial cells 6 (IEC-6) after heat stress, and explore the pathogenesis of intestinal damage caused by heat stress.Methods In the heat stress groups,the cells were incubated at 43℃ for 1 hour, then, further incubated at 37℃ and 5% CO2 for 0, 1, 3, 6 and 12 hours respectively; in the medicine intervention group, the cells were pretreated with the medicine 1h before heat stress; while in control group, the cells were incubated at 37℃ and 5% CO2. The amount of reactive oxygen species (ROS) was assayed with 2', 7'-dichlorofluorescin diacetate (DCFH-DA) and dihydroethidium (DHE) staining. The stability of lysosome membrane was checked by AO staining. Apoptosis was analyzed by flow cytometry using annexinⅤ-FITC/PI staining, CCK-8 assay was used to assess cellular viability.Results Compared with control group, cell viability decreased and apoptosis increased at 1 h after heat stress, which was the most obvious at 12h after rewarming (P<0.05). While ROS and pale cells increased immediately after heat stress and the increase become the most obvious (P<0.05). The cell viability in E-64 pretreatment group was significantly improved such as apoptosis reduction, compared with heat stress group (P<0.05).Conclusion Heat stress could induce robust increase of ROS, which mediates lysosome damage and results in cell apoptosis, thus suggesting that ROS-lysosome pathway may play an important role in intestinal injury in heat stress.

Objective To analyze the change of chromosome test level after participating in the inter‐laboratory proficiency test(PT) program of the College of American Pathologists (CAP) .Methods The results for participating in CAP PT during 2011-2014 were analyzed ,the accuracy of the cytogenetic chromosome quality control test in this laboratory was obtained accord‐ing to the CAP evaluation results ,and the effect of CAP external quality assessment item on increasing the chromosome detection level in this laboratory was evaluated by analyzing the chromosome band levels before and after participating in CAP .Results This laboratory participated in CAP PT test for 10 times during 2011-2014 ,a total of 59 cases were analyzed ,the accuracy rate for jud‐ging chromosome karyotype was 100% ,the karyotype description accuracy rate was 95 .1% .The chromosome test results of clinical cases in this laboratory displayed that peripheral blood chromosome abnormal detection rate was 18 .9% and bone marrow chromo‐some abnormal detection rate was 25 .9% ,the abnormal rate of newly diagnosed leukemia was 66 .8% ;the detection failure rates of peripheral blood chromosome and bone marrow chromosome were 0 .5% and 5 .0% respectively ;the detection failure rates of pe‐ripheral blood chromosome and bone marrow chromosome after participating in CAP were decreased ,the chromosome band average level was improved ,showing statistical difference compared with those before participating in CAP (P<0 .01) .Conclusion Partici‐pating in high quality external laboratory assessment item can increase the clinical analytic level of cytogenetic chromosome test .

Objective To investigate the variation and clinical significance of serum levels of inflammatory cyto-kines after percutaneous coronary intervention (PCI)treatment in acute myocardial infarction (AMI)patients. Methods 118 incipient AMI patients with successful underwent PCI (Defined as treatment group,blood samples were collected from pre-operation,12 h after operation,24 h after operation,48 h after operation and 90 d postop-erative follow-up period)and 52 AMI patients with diagnostic coronary angiography (CAG)(Defined as control group,blood samples were collected prior to CAG,12 h after CAG,24 h after CAG,48 h after CAG and 90 d fol-low-up period)were enrolled in this study.Serum levels of IL-6,IL-18,hs-CRP,TNF-αand MMP-9 were detec-ted in all the subjects by enzyme-linked immune sorbent assay(ELISA)and major adverse cardiac events(MACE) occurrence rate was analyzed in 90 days followed-up cases.Results No significant differences in baseline levels of IL-6,IL-18,hs-CRP,TNF-αand MMP-9 were found in the two study groups(P >0.05).No significant differences of levels of IL-6,IL-18,hs-CRP,TNF-αand MMP-9 were found after CAG in control group (P >0.05).The serum levels of IL-6,IL-18,hs-CRP and TNF-αafter PCI were significantly increased (P <0.01 )while no significant differences were found in level of MMP-9 (P >0.05)in PCI group.There were significant differences of levels of IL-6,IL-18,hs-CRP and TNF-αbetween MACE group and without MACE group after PCI.The multivariable lo-gistic analysis showed that IL-6,IL-18,hs-CRP and TNF-αwere risk factors of MACE after 90 days follow-up. Conclusion The concentrations of serum IL-6,IL-18,hs-CRP and TNF-αare significantly increased in AMI pa-tients treated with PCI.PCI operation may induce inflammatory reaction.High serum levels of peripheral inflamma-tory cytokines IL-6,IL-18,hs-CRP and TNF-αhave an important role in major adverse cardiac events(MACE)and short-term prognosis in the first AMI patients treated with successful primary PCI.

OBJECTIVETo analyze the results of prenatal karyotype of the external quality assessment program in 2013 in order to provide references and recommendations for improving the capability and performances of karyotype analysis of prenatal screening laboratories.

METHODSFive lots of quality control cell photos were sent to 500 laboratories. The participants were asked to decide whether the photos have demonstrated any abnormal karyotype and determine the abnormal type. The results should be submitted before the deadline and compared with the standard results to evaluate the performances of the laboratory.

RESULTSOne hundred forty three laboratories have returned their karyotype results for the survey. The standard answers were 7,XX,+18, 46,X,i(X)(q10), 46,XY,i(21)(q10) or 46,XY,+21,der(21;21)(q10;q10), 46,XY and 47,XY,+21 in sequential order, which were used to estimate the score of each participant. The pass rates for five lots were 97.9%, 97.2%, 95.8%, 100.0% and 97.9%, respectively. The total pass rate was 97.7%. The error rates were 2.1%, 2.8%, 4.2%, 0 and 2.1%, respectively. The total error rate was 2.3%.

CONCLUSIONSome laboratories did not correctly identify the abnormal karyotypes, while some could not determine the right type of karyotype. The external quality assessment program of prenatal diagnosis of karyotype analysis should be conducted annually in order to improve the capability and performances of karyotype analysis of prenatal screening laboratories.

Adult ; Chromosomes, Human ; genetics ; Female ; Fetal Diseases ; diagnosis ; genetics ; Genetic Testing ; methods ; standards ; Humans ; Karyotyping ; methods ; standards ; Male ; Pregnancy ; Prenatal Diagnosis ; methods ; standards ; Young Adult

Objective To investigate the optimal method for synthesizing thiolated doxorubicin .Methods Thiolated doxorubi-cin was synthesized through two different methods .Doxorubicin was reacted with 2-iminothiolane (2-IT) and S-acetylthioglycolic acid N-hydroxysuccinimide ester (SATA),respectively.The synthesized thiolated doxorubicin was further characterized by HPLC and MS -ESI techniques .Several factors including molar ratios as well as reaction time were evaluated .Results The results showed that thiolat-ed doxorubicin could be synthesized via both of the two methods successfully .Thiolated doxorubicin could be stable when doxorubicin was reacted with SATA .But the crude thiolated doxorubicin could be cyclized easily when doxorubicin was reacted with 2-IT.Conclu-sion Thiolated doxorubicin prepared with SATA is more feasible than that with 2-IT.

Objective To investigate clinical effect and safety of in vitro maturation(IVM)of human immature oocytes in infertile women with polycystic ovary syndrome by comparing with conventional in vitro fertilization(IVF)and intracytoplasmic sperm injection(ICSI).Methods From Jan.2003 to Dec.2009,157 infertile women with PCOS underwent 162 cycles IVM in Center for Reproductive Medicine,the First Affiliated Hospital of Anhui Medical University.In the mean time,109 patients with PCOS underwent 114 IVF/ICSI cycles as control group 1 and 106 patients with other factors underwent 106 IVF/ICSI cycles as control group 2.Treatment and outcome of pregnancy and infant were compared among those 3 groups.Results No statistically significant difference were found in terms of the positive rate of hCG in urine[35.7％(56/157),42.2％(46/109),44.3％(47/106)],the rate of clinical pregnancy[29.3％ (46/157),37.6％(41/109),41.5％(44/106)],the rate of entopic pregnancy[1.9％(3/157),1.8％ (2/109),0.9％(1/106)],the rate of miscarriage[18.6％(8/43),12.8％(5/39),20.9％(9/43)]and the rate of live-birth[22.3％(35/157),31.2％(34/109),32.1％(34/106)]among three grbups(IVM group,control group 1,control group 2,P ＞ 0.05).The rate of preterm labor,low weight newborn,mean birth weight,ratio of male to female did not show significantly difference among 3 groups(P ＞ 0.05).The average control ovarian stimulation was 6 days,the median dose of gonadotropin(Gn)was 675 IU,and the total hospital cost was(8392 ± 1328)RMB in IVM group,which were statistically lower than those in the other two control groups(P ＜ 0.01).The rate of multiple pregnancy was 4.7％(2/43)and ovarian hyperstimulation syndrome(OHSS)0 in IVM group,which were significantly lower than those in the other control group(P ＜0.01).Conclusion In vitro maturation is an effective treatment in infertile women with PCOS,it could obtain the similar pregnancy outcome and reduce total cost,the dosage of gonadotropinreleasing hormone and rate of OHSS compared with conventional IVF/ICSI.