Research progress on screening of pregnancy-related diseases based on fetal cell free DNA / 中华医学遗传学杂志

Fetal cell free DNA (cfDNA) in maternal blood circulation mainly originates from placental trophoblasts which have dual characteristics of apoptotic cells and the embryo, and can be affected by maternal factors. Pregnancy-related diseases including preeclampsia, gestational diabetes mellitus, preeclampsia, macrosomia and fetal growth restriction can seriously affect maternal health and pregnancy outcome. Early prediction and timely intervention are important means to reduce the risk. Fetal cfDNA and prediction of pregnancy-related diseases have become a hot topicfor current research. This paper reviews the latest progress made in the field.

Targeted elimination of mutant mitochondrial DNA in MELAS-iPSCs by mitoTALENs

Mitochondrial diseases are maternally inherited heterogeneous disorders that are primarily caused by mitochondrial DNA (mtDNA) mutations. Depending on the ratio of mutant to wild-type mtDNA, known as heteroplasmy, mitochondrial defects can result in a wide spectrum of clinical manifestations. Mitochondria-targeted endonucleases provide an alternative avenue for treating mitochondrial disorders via targeted destruction of the mutant mtDNA and induction of heteroplasmic shifting. Here, we generated mitochondrial disease patient-specific induced pluripotent stem cells (MiPSCs) that harbored a high proportion of m.3243A>G mtDNA mutations and caused mitochondrial encephalomyopathy and stroke-like episodes (MELAS). We engineered mitochondrial-targeted transcription activator-like effector nucleases (mitoTALENs) and successfully eliminated the m.3243A>G mutation in MiPSCs. Off-target mutagenesis was not detected in the targeted MiPSC clones. Utilizing a dual fluorescence iPSC reporter cell line expressing a 3243G mutant mtDNA sequence in the nuclear genome, mitoTALENs displayed a significantly limited ability to target the nuclear genome compared with nuclear-localized TALENs. Moreover, genetically rescued MiPSCs displayed normal mitochondrial respiration and energy production. Moreover, neuronal progenitor cells differentiated from the rescued MiPSCs also demonstrated normal metabolic profiles. Furthermore, we successfully achieved reduction in the human m.3243A>G mtDNA mutation in porcine oocytes via injection of mitoTALEN mRNA. Our study shows the great potential for using mitoTALENs for specific targeting of mutant mtDNA both in iPSCs and mammalian oocytes, which not only provides a new avenue for studying mitochondrial biology and disease but also suggests a potential therapeutic approach for the treatment of mitochondrial disease, as well as the prevention of germline transmission of mutant mtDNA.

Construction of vascular endothelial growth factor modified muscle-derived stem cells and its expression / 实用医学杂志

Objective To construct the lentiviral vector encoding vascular endothelial growth factor gene and detect the vascular endothelial growth factor (VEGF) expression in muscle-derived stem cells (MDSCs). Methods To culture MDSCs and detect the CD34,CD45,Bcl-2 and Desmin expression in MDSCs by immunofluorescence. A cDNA encoding VEGF gene was amplified by PCR. This fragment was cut with EcoRI and BamHI and ligated with an EcoRI- and BamHI-reated lentiviral vector pCDH-CMV-MCS-EF1-copGFP. Then DNA sequencing analysis was performed to confirm successful construction of pCDH-CMV-MCS-EF1-copGFP -VEGF. The expression of VEGF was confirmed using enzyme-linked immunosorbent assay (ELISA), Western blot, and Real-time PCR analyses. Results The pCDH-CMV-MCS-EF1-copGFP-VEGF lentiviral vector was constructed successfully. When MOI values in the transfection efficiency MDSCs by FCM. were 1,5,15, the transfection rate reached to 16.7%, 45.6%, 66.3% and 85.6% respectively. When MOI value was of 20, the rate was up to 90.1%. Real-time PCR, Western blot and ELISA showed stable expression of VEGF in MDSCs. Conclusion We successfully constructed lentiviral vector carrying the VEGF and stable expression in MDSCs.

Influence of male age on the pregnancy outcomes of IVF-ET / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the influence of male age on the pregnancy outcomes of in vitro fertilization-embryo transfer (IVF-ET).</p><p><b>METHODS</b>We retrospectively analyzed 7,533 cycles of IVF-ET performed between January 1, 2009 and October 31, 2013. We divided the samples into three groups according to the female age (< 30, 30-34, and 35-38 yr), each again subdivided into six groups based on the male age (< 30, 30-32, 33-35, 36-38, 39-41, and ≥ 42 yr). We compared the rates of implantation, pregnancy, miscarriage, and live birth among different age groups.</p><p><b>RESULTS</b>There were no statistically significant differences in basal E2, FSH, endometrium thickness on the day of hCG administration, number of oocytes retrieved, and days of embryo transfer among different male age groups (P > 0.05). The implantation rate showed an age-dependent decrease in the < 30, 30-32, 33-35, 36-38, 39-41, and ≥ 42 yr male groups, 41.1, 42.0, 39.5, 31.3, 40.7, and 48.6% among the women aged < 30 years (P < 0.05), 40.3, 36.4, 35.1, 35.3, 29.4, and 37.3% among the women aged 30-34 years (P < 0.05), and 48.2, 17.8, 25.3, 23.5, 22.1, and 23.8% among the women aged 35-38 years (P < 0.05). The miscarriage rate was significantly higher in the ≥ 39 yr than in the 30-32 and 33-35 yr male age groups among the women aged 30-34 years (P < 0.05), but showed no remarkable differences among the other male age groups in the women aged < 30 and 35-38 years (P > 0.05). No statistically significant differences were observed in the rates of pregnancy and live birth among different male age groups (P > 0.05).</p><p><b>CONCLUSION</b>Male age has some influence on the rates of implantation and miscarriage but not on the rates of pregnancy and live birth in IVF-ET.</p>

Percentage of uric acid calculus and its metabolic character in Dongjiang River valley / 中华外科杂志

<p><b>OBJECTIVE</b>To study the percentage of uric acid calculus in uroliths and its metabolic character in Dongjiang River valley.</p><p><b>METHODS</b>To analyze the chemical composition of 290 urinary stones by infrared (IR) spectroscopy and study the ratio changes of uric acid calculus. Uric acid calculus patients and healthy people were studied. Personal characteristics, dietary habits were collected. Conditional logistic regression was used for data analysis and studied the dietary risk factors of uric acid calculus. Patients with uric acid calculus, calcium oxalate and those without urinary calculus were undergone metabolic evaluation analysis. The results of uric acid calculus patients compared to another two groups to analysis the relations between the formation of uric acid calculus and metabolism factors.</p><p><b>RESULTS</b>Uric acid calculi were found in 53 cases (18.3%). The multiple logistic regression analysis suggested that low daily water intake, eating more salted and animal food, less vegetable were very closely associated with uric acid calculus. Comparing to calcium oxalate patients, the urine volume, the value of pH, urine calcium, urine oxalic acid were lower, but uric acid was higher than it. The value of pH, urine oxalic acid and citric acid were lower than them, but uric acid and urine calcium were higher than none urinary calculus peoples. Blood potassium and magnesium were lower than them.</p><p><b>CONCLUSIONS</b>The percentage of uric acid stones had obvious advanced. Less daily water intake, eating salted food, eating more animal food, less vegetables and daily orange juice intake, eating sea food are the mainly dietary risk factors to the formation of uric acid calculus. Urine volume, the value of pH, citric acid, urine calcium, urine uric acid and the blood natrium, potassium, magnesium, calcium, uric acid have significant influence to the information of uric acid stones.</p>