Mitochondrial disease is an unusual genetic disease .The in vitro fertilization technology of “three-parent embryos” as a new assisted reproductive technology , may help women to prevent their mitochondrial diseases passing on to their children , but it′s still controversial in many aspects .The article presents some scientific and ethical analysis and reflection on this technology .In terms of its necessity , safety, efficacy, and ethical were dis-cussed , and the application of the technology may bring the risk of concern is put forward , based on the present sit-uation of and attitude toward the technology should be taken to keep .

Objective To investigate the effects of downstream regulatory element antagonist modulator (DREAM) on the expression of glutamate transporter-1 (GLT-1) in spinal cord in rats with bone cancer pain and morphine tolerance.Methods Sixty female healthy Sprague-Dawley (SD) rats weighing 200 ～230 g were randomly divided into tow groups,group Ⅰ cancer pain (CP,n =48) and group Ⅱ Sham (S,n =12).Cancer pain in each group was produced by inoculation of syngenetic Walker 256 rat mammary gland carcinoma cells (5 × 105) to left tibia.Pain threshold to mechanical stimulus was measured before (baseline) and after the surgical procedure.From 14 d to 18 d after the inoculation of carcinoma cells,36 rats from group CP received subcutaneous injection of morphine at 3 times per day with doses increasingly from 10 mg/kg initially to 20 mg/kg,30 mg/kg,40 mg/kg,and 60 mg/kg.Equal volume of normal saline was applied to the 12 rats left in group CP.On 19th day after the carcinoma cells inoculation once subcutaneous injection of morphine at 3mg/kg was performed in all rats in group CP.From the next day,the rats in group CP ever receiving injections of morphine for 5 days were randomized into thre subgroups,including subgroups morphine tolerance (MT,n =12),vehicle (V,n =12),and RNAi (R,n =12).They were injected intrathecally with 20 μl of normal saline (NS),10 μl vehicle plus 10 μ1 NS,and 10 μ1 of DREAM-shRNA plus l0 μ1 NS,respectively,once a day for 5 days.Focusing on the affected limb,mechanical pain threshold was measured one day before surgery (T0),and at day7 (T1),day 14 (T2),day 18 (T3),day 19 (T4),day21 d (T5),day 25 (T6),and day 28 (T7) after surgery.The animals were sacrificed at day 28 after the procedure.The lumbar 4 segments in rats were removed for detection of DREAM and GLT-1.Results The mechanical threshold was significantly decreased at T1 compared to the baseline in all groups,returned to the baseline at T2 ～ T7 in group S,at T4 in group CP,and at T2 in group MT,V,and R,but remained low at T5 ～T7 in group CP,and at T3 ～T7 in group MT,V,and R.Compared to that at T1,it was decreased at T2 ～T3 and T5 ～ T7 in group CP,at T4 ～ T7 in group MT and V,and at T4 ～ T5 in group R,going back to the baseline at T4 in group CP and at T2 in group MT,V and R,and increased at T6 ～ T7 in group R.Compared to that in group S,the mechanical threshold in group CP,MT,V and R was decreased,and lower at T2 in group CP and at T4 in group MT,V and R.Compared to that in group CP,the mechanical threshold was significantly higher at T2 ～ T3 but lower at T4 in group MT,V,and R,decreased at T5 in group R and at T5 ～ T7 in group MT and V.The mechanical threshold was increased at T6 ～ T7 in group R and higher than that in groups MT and V.The expression of DREAM,compared to that in group S,was down-regulated in other groups.Compared to group CP,increment was shown in groups MT and V,and decrease was exhibited in group R.It was cut down in group R compared to that in groups MT and V.Compared to group S,GLT-1 was decreased in other groups.It was down-regulated in groups MT,V and R compared to group CP.Conclusions DREAM is involved in the development of allodynia after morphine tolerance in rats with bone cancer pain.No evidence in this study supports a link between DREAM and GLT-l in spinal cord.

Objective To investigate the association of the expressions of glomerular nephrin, vascular endothelial growth factor (VEGF) and its receptor (VEGFR) with proteinuria in preeclampsia rats. Methods A rat model of preeclampsia was developed by inhibitor of nitric oxide synthase (L-NAME). The systolic blood pressure (SBP) and 24 h urine protein were compared among the normal female group (n=6), the normal pregnant group (n=8), nonpregnant control group (n=6) and preeclampsia group(n=8). The kidney biopsies of each group were observed by light and electron microscopy. The glomerular nephrin was detected by Western blotting and real-time PCR. Immunofluorescence was used to detect the expression of WT1. The level of glomerular VEGF and VEGFR (Flt-1 and Flk-1) were evaluated by Western blotting. Results The level of glomerular nephrin protein in the rats with preeclampsia (0.0726±0.0074) was significantly lower compared with normal female group (0.3795±0.0509), normal pregnant group (0.2361±0.0437) and nonpregnant control group (0.7265±0.0503) (P＜0.01, respectively), while the levels of nephrin mRNA were not significantly different among 4 groups. The expression of WT1 was not significantly different among 4 groups as well. The level of glomerular VEGF in preeclampsia group (1.5429±0.0898) was significantly higher compared with normal female group (1.1870±0.1160), normal pregnant group (1.3741 ±0.1165) and nonpregnant control group (1.0155±0.0742)(P＜0.01,respectively). VEGFR (Flt-1 and Flk-1) was also significantly higher in preeclampsia rats compared with other control groups (P＜0.05, respectively). Conclusions In preeclampsia rats, nephrin is decreased significantly and the glomerular VEGF-VEGFR is increased significantly compared with the other control groups. The abnormal expression of nephrin and VEGF-VEGFR may be involved in the preeclampsia proteinuria. The underlying mechanism of this phenomenon needs further research.

Objective To explore the impacts of preeclampsia and the different extent of proteinuria on maternal and perinatal outcomes.Methods The retrospective analysis was conducted according to the perinatal clinical data of preelacmpsia,pregnancy-induced hypertension in pregnant women and normal pregnant women from the Fifth People's Hospital of Shanghai,excluding twins,diabetic mellitus and patients with chronic kidney disease previously.Patients were divided into three groups on the basis of their conditions:① preeclampsia patients (A group,220 cases); ② patients with gestational hypertension (B group,189 cases); ③ normal pregnant (C group,220 cases).Patients with pre-eclampsia according to the degree of proteinuria were further divided into three subgroups:A1:patients with mild proteinuria (n =109); A2:patients with moderate proteinuria (n =72); A3:patients with severe proteinuria (n =39).Results Compared with the other two groups,the patients in A group had higher blood pressure,serum creatinine,uric acid,cesarean section rate,perinatal prematurity,stillbirth,fetal distress and neonatal asphyxia in preeclampsia group.However,the serum albumin level,eGFR,neonatal birth weight,length and Apgar scores were lower in A group compared with B and C group (P ＜ 0.05).In three subgroups,serum creatinine level,uric acid level,cesarean section rate,perinatal prematurity and fetal distress were significantly increased in A3 group compared with A1 group,while the serum albumin level,eGFR,gestational age and neonatal birth weight were obviously lower in A3 group than in A1 group (P ＜ 0.05).In patients with preeclampsia,24 h urinary protein was negatively related with the levels of serum albumin and eGFR (P ＜ 0.05),and positivly related with the blood pressure,serum creatinine and caesarean production rate (P ＜ 0.05).Large amounts of proteinuria was a risk factor of adverse outcome for pregnant patients with preeclampsia (OR =2.899,P ＜ 0.05).Conclusions Preeclampsia patients with large amount of proteinuria have poor maternal and perinatal outcomes.Massive proteinuria is a risk factor of adverse outcome for patients with pre-eclampsia.

Objective To evaluate the effectiveness and safety of acupuncture in the management of post-stroke spasticity.Methods Six domestic and abroad databases were screened by computer.Randomized controlled trials (RCT)were included if acupuncture treatment was used as the sole intervention or as an adjunction to another conventional treatment on post-stroke spasticity. Two independent reviewers screened the articles,extracted data and assessed the risk of bias.The evaluation of the quality was performed using Cochrane Risk of Bias Tool and the data was analyzed by using RevMan 5.3.Mean difference (MD)and 95% confidence interval (CI)were used to describe quantitative data.Results A total 29 RCT were included,7 of them had high risk of bias and the rest of them had unclear risk of bias. Acupuncture was better than rehabilitation therapy in remission of spasticity (-0.40 [-0.68,-0.12]).The 4 or 8 weeks of treatment in studies of unmentioned spasmodic regions showed that acu-puncture with rehabilitation therapy was better than rehabilitation therapy (-0.31 [-0.50,-0.11],-0.41[-0.54,-0.28]).The studies of 4 weeks of treatment showed superior effect of acupuncture with conventional treatment compared with rehabilitation therapy with conventional treatment (-0.96 [-1.73,-0.19]),and acupuncture with rehabilitation therapy was superior to conventional treatment with rehabilitation therapy (-0.57[-0.80,-0.34]).After 6 months of treatment,acupuncture with nerve block therapy and rehabilitation therapy was better than nerve block therapy with rehabilitation therapy (-0.63[-1.12,-0.14]),and acupuncture with nerve block therapy was better than nerve block therapy (-0.70[-1.08,-0.32]).In improving the physical activity,acupuncture,acupunc-ture with rehabilitation therapy,acupuncture with conventional treatment and rehabilitation therapy 1 or 4 weeks of follow-up,and acupuncture with nerve block therapy was better than that of control group. Conclusion Acupuncture could remiss the spasticity after stroke and improve the ability of daily life and motor function of patient,but the conclusion of subgroups are not the same.The high quality RCT studies with clear spasmodic position and measurement at more time points are needed to confirm the effectiveness and safety of acupuncture in post-stroke patients with spasticity.

Background and purpose: Abnormal expression and amplification of transforming growth factor beta 1 (TGF-β1) and Notch3 in ovarian carcinoma tissues are associated with metastasis and low survival rate, respectively. The crosstalk between TGF-β1 and Notch3 signaling pathway promotes invasion and metastasis in various cancers. However, the mechanism is still under debate. Therefore, this study was designed, using in vitro cytological assays, to investigate the effects of TGF-β1 and Notch3 signaling pathway on ovarian cancer cell biological behavior and the potential mechanisms in terms of the crosstalk between TGF-β1 and Notch3 signaling pathway. Methods: Hey A8 and Hey cell lines were used as models in the study. The levels of TGF-β1 in supernatants from culture media were measured by ELISA. Both cell lines were treated with 500 ng/mL TGF-β1 neutralizing antibody (control group), 10 ng/mL TGF-β1, 50 μmol/L DAPT, 10 ng/mL TGF-β1 and 50 μmol/L DAPT, 50 μmol/L tumor necrosis factor receptor-associated factor 6 (TRAF6) peptide inhibitor, 10 ng/mL TGF-β1 and 50 μmol/L TRAF6 peptide inhibitor, respectively. The protein expression levels of TGF-β1 and Notch3 signaling pathway molecules as well as TRAF6 from cell lines with different treatments were detected by Western blot. Cell proliferation, migration and invasion were tested by cell counting kit-8 (CCK-8), scratch and Transwell assays, respectively. Results: The levels of TGF-β1 were timedependently increased in supernatants of culture media from Hey A8 and Hey cell lines. Compared with control group, TGF-β1 treatment increased the expression levels of Notch3-ICD and Hes1, while no obvious change was observed in the group treated with DAPT and TGF-β1. Moreover, TGF-β1 promoted cell proliferation, migration and invasion while DAPT decreased the proliferation, migration and invasion in cell lines treated with TGF-β1. These results indicated that TGF-β1 might promote proliferation, invasion and migration of ovarian epithelial cancer cells through activating the Notch3 signaling pathway. Further study showed that TGF-β1 up-regulated TRAF6 and activated the Notch3 signaling pathway. The activation of the Notch3 signaling pathway by TGF-β1 was inhibited in cells treated with the TRAF6 specific inhibitor. Conclusions: TGF-β1 may promote the proliferation, invasion and migration of ovarian epithelial carcinoma cells through TRAF6-mediated activation of the Notch3 signaling pathway.

Objective:To explore the efficacy and safety of anti-tumor necrosis factor alpha (TNFα) monoclonal antibodies (mAbs) for severe/refractory vasculo-Behcet′s disease (BD).Method:The clinical data of severe/refractory vasculo-BD patients treated with anti-TNFα mAbs were retrospectively analyzed. Response of anti TNFα mAbs was analyzed. The dosage changes of glucocorticoid, the level of erythrocyte sedimentation rate (ESR) and hypersensitive C-reactive protein (hsCRP) before and after treatment were recorded, as well as side effects.Result:Sixteen patients were enrolled. Arterial lesions were reported in 12 patients, including 9 with arterial aneurysm, 6 with arterial dilation, 2 with stenosis and 2 with occlusion. Seven patients presented venous thrombosis, including lower extremity veins ( n=6), cerebral venous sinus ( n=2) and inferior vena cava system ( n=2). Two cases had both arterial and venous involvement. Before the application of TNFα mAbs, all 16 patients failed to response to prednisone or its equivalent dose of 40 (7.5-90) mg/d in combination with cyclophosphamide, methotrexate, thalidomide or azathioprine for median 4 (0-156) months. After a mean duration of treatment for (17.1±6.5) months, 15 patients achieved complete remission and 1 patient achieved partial remission. Three patients received surgery without any postoperative complications. After using anti TNFα mAbs, the dosage of prednisone [5(0-12.5)mg/d vs. 40(7.5-90)mg/d, P<0.01], ESR [(7.3±4.6) mm/1h vs. (33.5±26.7) mm/1h, P<0.01] and hsCRP [1.9(0.2-11.4) mg/L vs. 24.3(0.4-113.9) mg/L, P<0.01] were significantly decreased. Side effects were observed in 2 patients. One developed pulmonary infection 12 months after adalimumab with conventional treatment. Another patient had allergy to infliximab then switched to adalimumab. Conclusion:In combination with corticosteroids and immunosuppressants, anti-TNF α mAbs are effective and well-tolerated in severe/refractory vasculo-BD, with a favorable steroid -sparing effect and rare postoperative complications.

New threats posed by the emerging circulating variants of SARS-CoV-2 highlight the need to find conserved neutralizing epitopes for therapeutic antibodies and efficient vaccine design. Here, we identified a receptor-binding domain (RBD)-binding antibody, XG014, which potently neutralizes β-coronavirus lineage B (β-CoV-B), including SARS-CoV-2, its circulating variants, SARS-CoV and bat SARSr-CoV WIV1. Interestingly, antibody family members competing with XG014 binding show reduced levels of cross-reactivity and induce antibody-dependent SARS-CoV-2 spike (S) protein-mediated cell-cell fusion, suggesting a unique mode of recognition by XG014. Structural analyses reveal that XG014 recognizes a conserved epitope outside the ACE2 binding site and completely locks RBD in the non-functional "down" conformation, while its family member XG005 directly competes with ACE2 binding and position the RBD "up". Single administration of XG014 is effective in protection against and therapy of SARS-CoV-2 infection in vivo. Our findings suggest the potential to develop XG014 as pan-β-CoV-B therapeutics and the importance of the XG014 conserved antigenic epitope for designing broadly protective vaccines against β-CoV-B and newly emerging SARS-CoV-2 variants of concern.
Angiotensin-Converting Enzyme 2 ; Antibodies, Neutralizing ; Antibodies, Viral ; COVID-19 ; Epitopes ; Humans ; SARS-CoV-2/genetics* ; Spike Glycoprotein, Coronavirus/genetics*