Objective:To investigate the effect of in vitro maturation(IVM),fertilization and embryo transfer of immature oocytes derived from nature cycles in infertility couples.Methods:30 IVF/ICSI-ET cycles were involved in this study.Oocytes retrieval performed after follicle larger than 10 mm and before endogenesis Luteinizing hormone(LH) surge occurrence in nature cycle.Mature oocytes(Metaphase Ⅱ) were inseminated by standard in vitro fertilization(IVF) on the oocytes pick-up day.Immature oocytes(Metaphase I and germinal vesicle stage) were cultured for 24~48 hours and fertilized by intracytoplasmic sperm injection(ICSI) as soon as it developed to Metaphase Ⅱ stage.Embryo transfer was performed 72 h after fertilization.Results:2 of 30 IVM cycles brought no oocytes in this study.113 oocytes were obtained among other 28 oocyte retrieval cycles,74 of which were immature,29 were mature,and 10 were degenerated.60 of 74 immature oocytes developed to MⅡ after IVM(maturation rate 81.08%).6 cases of clinical pregnancy were obtained in the 20 embryo transfer cycles,five of which brought healthy babies(one came from immature oocyte and four came from mature oocyte) and one was abortive.Conclusion:IVM,fertilization and embryo transfer of immature oocytes derived from nature cycles is a feasible treatment for women with infertility.

AIM:To study the expression of Jagged 2/Notch3 signaling molecules in pulmonary vascular wall of pulmonary hypertensive rats induced by monocrotaline .METHODS: SD rats were randomly divided into normal control group (C group,n=15), solvent control group (S group,n=15) and monocrotaline model groups (M group,n=15).The model of pulmonary hypertension was established by a single subcutaneous injection of monocrotaline (50 mg/kg).The rats in S group were given a single subcutaneous injection of the same dose of solvent .After 4 weeks, the pulmonary vascular remodeling was assessed by HE staining , and the mean pulmonary artery pressure ( mPAP) and right ventricular systolic pressure (RVSP) were determined by right heart catheterization .The expression of Jagged2/Notch3 /Hes5 molecules in the pulmonary vascular wall was detected by immunohistochemical method and real -time PCR.RESULTS:Compared with S group and C group , the percentage of medial wall thickness of smaller arteries in model group increased significantly (P<0.01).The levels of mPAP and RVSP in M group were significantly higher than those in S group and C groups (P<0.01).The results of real-time PCR showed that the expression of Jagged 2, Notch3 and Hes5 was significantly increased in M group compared with S group and C group .The data from immunohistochemical detection indicated that Jagged 2 mainly expressed in the intima of small lung artery , Notch3 and Hes5 mainly expressed in the medial smooth muscle cells .Com-pared with S group and C group , the expression of Jagged 2 and Notch3 was significantly increased in the lung small arteries of M group.CONCLUSION:The activation of Jagged2/Notch3 signaling pathway might play an important role in the for-mation of pulmonary hypertension .

The elasticity of the ascending aorta in healthy volunteers and hypertension patients were examined by using quantitative tissue velocity imaging (QTVI), and the age-related change in the ascending aortic elasticity was investigated. The anterior and posterior walls of the ascending aorta were imaged with tissue Doppler method in all the subjects and QTVI was performed. Stable curves were obtained from 173 hypertension patients and 185 healthy adults. The peak early diastolic velocity (V(e)), peak late diastolic velocity (V(a)) and peak systolic velocity (V(s)) were measured. The relation of age with these measures was assessed. The results showed that the elasticity of the ascending aorta was much lower in the hypertension patients than in normal controls (P<0.05), and the elasticity was decreased with age in both groups (P<0.05). Our results suggested that QTVI, a new non-invasive ultrasonic technique, is helpful for the assessment of the aortic elasticity in hypertension patients.

Objective To observe the effect of platelet derived growth factor receptor β (PDGFR-β) transfected endothelial progenitor cells (EPCs) on vascular regeneration.Methods Spleen-derived mononuclear cells (MNCs) were isolated using density gradient centrifugation and induced with special culture medium.EPCs transfection was performed with LipofectamineTM 2000 reagent according to the instruction manual.Carotid artery injury was induced in splenectomized mice.EPCs were injected by tail vein immediately and at 24 h after endothelial injury of the carotid artery.Evans blue staining was performed to evaluate reendothelialization at 7 days after endothelial injury of the carotid artery.Results Most adherent cells were LDL and UEA-I double positive.Laser scanning confocal microscopy showed that transfection efficiency was about 50％-60％.The reendothelialized area in the PDGFR-β-EPCs group was significantly larger than that in EGFP-EPCs group.Conclusion Transplantation of PDGFR-β over-expressed EPCs can promote reendothelialization in the early phase after carotid artery injury.

OBJECTIVETo clone a novel human connexin gene and find out the relationship between this gene and hereditary deafness.

METHODSThrough the basic local alignment search tool (BLAST) analysis against the database of expressed sequence tags (dbEST) of National Center for Biotechnology Information (NCBI) using the coding sequence of mouse Cx 57 gene, 9 novel ESTs were obtained and a contig was assembled. Nested polymerase chain reaction (PCR) and rapid amplification of cDNA ends (RACE) were performed using primers designed on the contig. A novel gene was obtained and was mapped by homologous analysis against human genome sequence. Mutation analysis was performed in 12 autosomal dominant hereditary deafness families.

RESULTSNine ESTs were obtained by homologous analysis and a contig was assembled. Through nested PCR and RACE, a full length of cDNA was obtained from human liver, kidney Ready cDNA and placenta cDNA library, and was named CX 58. By comparison with human genome sequence, CX 58 was mapped at 1p32.3-p34.1. Mutation analysis of CX 58 was performed in 12 autosomal dominant hereditary deafness families, but no mutation was detected.

CONCLUSIONA novel human connexin gene named CX 58 was cloned and mapped to 1p32.3-p34.1. The mutation of CX 58 may not result in autosomal dominant hereditary deafness.

Animals ; Chromosome Mapping ; Chromosomes, Human, Pair 1 ; genetics ; Cloning, Molecular ; Connexins ; genetics ; DNA, Complementary ; chemistry ; genetics ; Expressed Sequence Tags ; Female ; Humans ; Mice ; Molecular Sequence Data ; Sequence Alignment ; Sequence Analysis, DNA

The elasticity of the ascending aorta in healthy volunteers and hypertension patients were examined by using quantitative tissue velocity imaging (QTVI),and the age-related change in the ascending aortic elasticity was investigated. The anterior and posterior walls of the ascending aorta were imaged with tissue Doppler method in all the subjects and QTVI was performed. Stable curves were obtained from 173 hypertension patients and 185 healthy adults. The peak early diastolic velocity (Ve),peak late diastolic velocity (Va) and peak systolic velocity (Vs) were measured. The relation of age with these measures was assessed. The results showed that the elasticity of the ascending aorta was much lower in the hypertension patients than in normal controls (P＜0.05),and the elasticity was decreased with age in both groups (P＜0.05). Our results suggested that QTVI,a new non-invasive ultrasonic technique,is helpful for the assessment of the aortic elasticity in hypertension patients.

Objectives: To provide new ideas on how to shift students' learning attitude from passive learning to active learning, we explored and evaluated a case/problem-based and interactive teaching mode in pathophysiology curriculum. Methods: Case/problem-based and interactive teaching mode is an innovative teaching model adopted in pathophysiology curriculum for grade 2015 students of 5-year program in clinical medicine and other medical students of non-clinical majors in Xiangya Medical School, Central South University. The teaching effectiveness of the case/problem-based and interactive teaching mode was evaluated by questionnaire survey, with 460 medical students enrolled in the survey whose approval degree on current teaching mode was analyzed. Excel was used to collect and process data, complete descriptive analysis and calculation of the percentage of indicators. Results: A total of 460 anonymous questionnaires were distributed and 453 valid questionnaires were retrieved, from which the following information was obtained: ① Pre-class learners' guidance designed for current teaching mode: 88.7% of students (402/453) believed that "Pre-class Learners' Guidance" motivated them to preview relevant teaching contents before class. 82.8% of students (375/453) believed "Pre-class Learners' Guidance" improved discussion quality in class. 76.6% of students (347/453) believed "Pre-class Learners' Guidance" expanded thinking and exploring space, while it did not increase student study burden (306/453, 67.6%). ② Compared with traditional teaching mode, the case/problem-based and interactive teaching mode had following advantages: It's helpful to cultivate students' clinical thinking (414/453, 91.4%), strengthen students' memory and understanding during study (400/453, 88.3%), attract students' attention in class (380/453, 83.9%), and aroused student's interest in class discussion (327/453, 72.2%). ③ 83.4% of students (379/453) preferred current teaching mode: they believed this teaching mode could improve students' ability to analyze and solve problems (325/453,71.7%), train clinical thinking (321/453, 70.9%), improve students' self-study ability (247/453, 54.5%) and increase students' capabilities of making summary and conclusion (197/453, 43.5%). Conclusion Case/problem-based and interactive teaching mode in pathophysiology curriculum enhances students' ability of self-studying, activates classroom's atmosphere, improves teaching quality, and effectively fosters students' clinical thinking. Therefore, this teaching mode deserves to be spread and applied in classroom teaching of pathophysiology and other basic medicine disciplines as well.

BACKGROUND:Myelodysplastic syndrome has worse hazards of acute myeloid leukemia transformation,and some studies have revealed that immune infiltration plays a vital part in the two.Nevertheless,more studies are required to confirm the relationship between immune infiltration and related differentially expressed gene regulation. OBJECTIVE:To screen the differentially expressed genes with prognostic significance between myelodysplastic syndrome and acute myeloid leukemia by bioinformatics analysis and explore the possible roles and mechanisms among these differentially expressed genes and immune infiltration mechanisms in the occurrence and progression of diseases. METHODS:The differentially expressed genes were screened for bioinformatics analysis using the GEO datasets,and analyzed by DO,GO,KEGG and GSEA.The TCGA prognostic database was used to plot the K-M curves of differentially expressed genes and receiver operating characteristic curve analysis was applied to evaluate the clinical diagnostic performance.Finally,CIBERSORT analysis was used to intuitively demonstrate the correlation between critical prognostic genes and the distribution of immuno-infiltrated cells.RT-qPCR was employed to detect peripheral blood samples from healthy controls,myelodysplastic syndrome and acute myeloid leukemia patients so as to verify the crucial genes preliminarily. RESULTS AND CONCLUSION:(1)A total of 150 differentially expressed genes were obtained between myelodysplastic syndrome and acute myeloid leukemia,among which 16 genes were up-regulated and 134 were down-regulated.(2)The results of DO,GO,KEGG and GSEA analysis suggested that differentially expressed genes might promote the development of myelodysplastic syndrome to acute myeloid leukemia by regulating the immune response.CIBERSORT revealed the differences in immune infiltration between myelodysplastic syndrome and acute myeloid leukemia.The distribution of CD4+ T cells,monocytes,neutrophils and M1 macrophages decreased in acute myeloid leukemia patients.In contrast,the distribution of inflammatory suppressor cells M2 macrophages increased,suggesting that it may be related to the immunosuppression of acute myeloid leukemia.(3)K-M curve and receiver operating characteristic curve analysis of 150 differentially expressed genes screened out four genes relevant to immunity and prognosis with good diagnostic performance:MANSC1,FLT3,BMX and CXCR2.(4)The results of RT-qPCR exhibited that MANSC1,BMX and CXCR2 were low expressed,while FLT3 was highly expressed in acute myeloid leukemia patients.These findings verify that the differential expression of MANSC1,FLT3,BMX and CXCR2 in patients with myelodysplastic syndrome and acute myeloid leukemia is not only significantly correlated with the prognosis of patients but may also affect the occurrence and development of myelodysplastic syndrome and acute myeloid leukemia by regulating the immune infiltration of patients.They can be used as potential biomarkers and therapeutic targets of the transformation from myelodysplastic syndrome to acute myeloid leukemia,providing a new direction for clinical diagnosis and treatment of the transformation of myelodysplastic syndrome.

To investigate whether intrauterine injection of human chorionic gonadotropin (hCG) before the embryo transfer in a frozen-thawed transfer cycle can improve the pregnancy outcome in the patients with repeated implantation failure (RIF).
 Methods: Prospective randomized-controlled trial was adopted. A total of 140 patients, who underwent thawed embryo transplantation and were in line with the diagnosis of RIF, were included. Other patients with some factors, such as uterine malformation, postoperative uterine cavity sticking, tubal effusion, endocrine diseases and endometriosis, were excluded. The patients were randomly divided into 2 groups through the computer random number table: an hCG intrauterine perfusion group and a control group. There was no significant difference in the age, the estradiol level, the number of transplanted embryos, the number of optimal embryos, and the thickness of the endometrium before transplantation between the 2 group (all P>0.05). The hCG+G2 fluid and the G2 fluid were prepared on the day of embryo transfer, and 40 μL of which was injected at an intrauterine site at 3 minutes before embryo transfer in the hCG intrauterine perfusion group and the control group, respectively. The clinical pregnancy rate and implantation rate in the 2 groups were compared.
 Results: The implantation rate and the clinical pregnancy rate in the hCG intrauterine perfusion group were higher than those in the control group (both P<0.05).
 Conclusion: The intrauterine injection of hCG can improve the implantation rate and pregnancy rate in cryopreserved embryo transfer in patients with RIF.
Chorionic Gonadotropin ; Embryo Implantation ; Embryo Transfer ; Female ; Humans ; Pregnancy ; Pregnancy Outcome ; Prospective Studies

BACKGROUND:U937 cells can be used as a cell model for studying the biological characteristics,signaling pathways,and therapeutic targets of acute myeloid leukemia.Although it has been reported that long non-coding RNA KIAA0125 is highly expressed in acute myeloid leukemia,its biological function in U937 cells remains unclear,and its mechanism of action in the occurrence and development of acute myeloid leukemia needs to be further clarified. OBJECTIVE:To investigate the expression level of long non-coding RNA KIAA0125 in peripheral blood of patients with acute myeloid leukemia and its effect on the proliferation and apoptosis of U937 cells. METHODS:RNA-sequencing was used to analyze the bone marrow monocyte samples from acute myeloid leukemia patients,and the differentially expressed gene long non-coding RNA KIAA0125 was screened.The expression of long non-coding RNA KIAA0125 in peripheral blood of patients with acute myeloid leukemia was detected by qRT-PCR.The relationship between long non-coding RNA KIAA0125 mRNA expression and prognosis in bone marrow cells of 173 acute myeloid leukemia patients and 70 healthy people was statistically analyzed by GEPIA database.Subsequently,recombinant lentivirus technology and CRISPR/Cas9-SAM technology were used to construct U937 cell lines with knockdown/overexpression of long non-coding RNA KIAA0125.qRT-PCR was used to detect the knockdown/overexpression efficiency of long non-coding RNA KIAA0125.Next,CCK-8 assay,flow cytometry,and western blot assay were used to detect the effects of knockdown/overexpression of long non-coding RNA KIAA0125 on the proliferation and apoptosis of U937 cells.Finally,western blot assay was used to detect the effect of knockdown/overexpressed long non-coding RNA KIAA0125 on Wnt/β-catenin signaling pathway-related proteins. RESULTS AND CONCLUSION:(1)The results of qRT-PCR showed that long non-coding RNA KIAA0125 was highly expressed in peripheral blood of acute myeloid leukemia patients.The results of GEPIA database showed that long non-coding RNA KIAA0125 was highly expressed in bone marrow cells of acute myeloid leukemia patients,and the high expression group had worse overall survival.(2)The knockdown efficiency of long non-coding RNA KIAA0125 in knockdown group was 70%,and the U937 cells that stably down-regulated long non-coding RNA KIAA0125 expression were successfully constructed.The expression of long non-coding RNA KIAA0125 in overexpression group was four times that of vector group,and stable U937 cells were successfully constructed.(3)Knockdown of long non-coding RNA KIAA0125 inhibited the proliferation of U937 cells and promoted their apoptosis.Overexpression of long non-coding RNA KIAA0125 promoted the proliferation of U937 cells but had no significant effect on the apoptosis of U937 cells.(4)Knockdown of long non-coding RNA KIAA0125 inhibited the activity of Wnt/β-catenin signaling pathway,while overexpression of long non-coding RNA KIAA0125 activated Wnt/β-catenin signaling pathway.These results confirm that long non-coding RNA KIAA0125 is highly expressed in acute myeloid leukemia peripheral blood.Long non-coding RNA KIAA0125 may affect the proliferation and apoptosis of U937 cells by regulating the Wnt/β-catenin signaling pathway,and may be a potential prognostic marker for acute myeloid leukemia.