We describe a genetic transformation method of secondary somatic embryogenesis in alfalfa through cotyledon-stage somatic embryos of alfalfa infected by Agrobacterium strain GV3101. The Agrobacterium strain GV3101 contained binary vector pCAMBIA2301 that had gus gene as reporter and npt II gene as selectable marker. The infected primary embryos were induced through series of medium under 75 mg/L kanamycin selection. We obtained the transgenic alfalfa plants. Then, GUS expression in different tissue of transgenic alfalfa was tested by GUS histochemical analysis. Further, the stable integration and transformation efficiency were tested by polymerase chain reaction and Southern blotting hybridization. The result showed that GUS expression was different in different organs of transgenic alfalfa; the copy number of integrated npt II gene was from 1 to 4; the transformation efficiency via primary somatic embryogenesis was 65.82%.
Agrobacterium ; genetics ; Medicago sativa ; embryology ; genetics ; physiology ; Plant Somatic Embryogenesis Techniques ; Plants, Genetically Modified ; embryology ; genetics ; Tissue Culture Techniques ; Transformation, Genetic

Objective:To evaluate the clinical efficacy and safety of fractional CO 2 laser for the treatment of vaginal relaxation syndrome. Methods:Twenty-one females with vaginal relaxation syndrome receiced between December 2016 and January 2017 in our department were treated for 3 times at 1 month interval with femilift fractional CO 2 laser. Vaginal palpation, vaginal squeeze pressure test, vaginal ultrasound, pelvic floor MRI, and Incontinence Questionnaire Short-Form (ICIQ-SF) assessments were performed at baseline and at post-treatment. Results:Significant improvement in female sexual dysfunction and stress urinary incontinence was especially seen based on patient＇s feelings after treatments. A noted progressive differences were observed in FSFI and vaginal pressure test, which were much higher after treatment than before treatment ( P<0.01). Moreover, in 13 cases suffering from mild to moderate stress urinary incontinence (SUI), fractional CO 2 laser treatment was associated with a significant improvement of ICIQ-SF scores ( P<0.01). All subjects successfully completed the study with no adverse events such as infections, bleeding and pain, etc. Conclusions:Fractional CO 2 laser treatment is an effective treatment for female sexual dysfunction and stress urinary incontinence caused by vaginal relaxation syndrome.

Objective To explore the relationship between fragmented QRS complexes (fQRS)and imperfect ST-segment resolution in ST elevation myocardial infarction (STEMI) patients undergoing primary percutaneous coronary intervention (p-PCI).Methods This study included 227 consecutive patients with STEMI who underwent p-PCI.They were divided into two groups:ECG with fQRS (n =142)and without fQRS (n =85).Baseline clinical characteristics,Gensini score,coronary angiography features and the rate of ST-segment resolution were compared between the two groups.Results (1) Patients with fQRS of ECG had higher cTnI,CK,CK-MB levels and Gensini score,prolonged QRS interval,lower rate of ST-segment resolution and left ventricular ejection fraction (LVEF) than in patients without fQRS (all P ＜ 0.01 or P ＜ 0.05).(2) Pearson correlation analysis showed that the rate of ST-segment resolution (r =-0.207,P =0.002),Gensini score (r =0.191,P =0.004),LVEF (r =-0.188,P =0.006),cTnI (r =0.172,P =0.010),and the TIMI grade post p-PCI (r =-0.148,P =0.028) were significantly related with the presence of fQRS.(3) Multivariate logistic regression analysis demonstrated that presence of fQRS at pre-PCI (OR =2.908,95％ CI:1.095-7.723,P =0.032),the number of leads with fQRS before PCI (OR =1.582,95％ CI:1.250-2.002,P ＜ 0.001),and increased QRS interval (OR =0.955,95％ CI:0.924-0.988,P =0.008) were independent predictors of imperfect ST-segment resolution.Conclusions fQRS is related to imperfect ST-segment resolution in STEMI patients undergoing p-PCI.fQRS may be a useful parameter to identify the patients with severe coronary lesion,larger areas of ischemic injury and myocardial infarction as well as severe left ventricular contracted dysfunction.

Objective:To explore the value of Gestational Diabetes Mellitus Risk Score in monitoring hypoglycemia among neonates delivered by mothers with gestational diabetes mellitus (GDM) .Methods:From February 2018 to June 2020, 240 neonates delivered by mothers with GDM admitted to the Shanghai First Maternity and Infant Hospital affiliated to Tongji University were selected as the study subject. The patients were assessed and divided into groups by the Gestational Diabetes Mellitus Risk Score and the Supplementary Gestational Diabetes Mellitus Risk Score from 12 to 20 weeks. The low-risk group ( n=63) was defined as the score of Gestational Diabetes Mellitus Risk Score ≤ 20 and the score of Supplementary Gestational Diabetes Mellitus Risk Score<33 from 12 to 20 weeks. The group with the score of Gestational Diabetes Mellitus Risk Score>20 and the score of Supplementary Gestational Diabetes Mellitus Risk Score <33 from 12 to 20 weeks was classified as the medium-risk group ( n=49) . High-risk group ( n=128) was defined as the score of Gestational Diabetes Mellitus Risk Score >20 and the score of Supplementary Gestational Diabetes Mellitus Risk Score >33 from 12 to 20 weeks. Three groups of patients were tested for fasting blood glucose (FPG) . Within one hour after birth, neonates were tested for blood glucose using the glucose oxidase blood glucose meter and given early sucking. Within 72 hours after birth, continuous dynamic blood glucose monitoring methods were used to measure blood glucose in neonates. This study compared the blood glucose within one hour and at 6, 12, 24, 48, and 72 hours after birth and the incidence of hypoglycemia at 72 hours after birth among three groups. Results:Among the 240 neonates in the risk group, 197 experienced hypoglycemia, with an incidence rate of 82.08%. There were statistically significant differences in the incidence of hypoglycemia before treatment, one occurrence of hypoglycemia within 72 hours, and recurrent hypoglycemia among the three groups of neonates ( P<0.01) . There were statistically significant differences in blood glucose among the three groups of newborns within one hour, 6, 12, and 24 hours after birth ( P<0.05) . There was no statistically significant difference in blood glucose among the three groups at 48 hours and 72 hours after birth ( P>0.05) . Conclusions:Neonates with gestational diabetes will suffer from repeated hypoglycemia. The higher the risk level, the higher the incidence of hypoglycemia. It is important to assess the risk of neonates with GDM, monitor their blood glucose in real time and treat them accordingly.

Objective:To investigate and compare the effects of water extracts of Whitmania pigra Whitman and Hirudinaria ma-nillensis Lesson on the angiogenesis of Tg (kdrl:mCherry) zebrafish. Methods:The zebrafish embryos 6-8 hours after fertilization (6-8hpf) were transferred to the culture medium containing Whitmania pigra Whitman or Hirudinaria manillensis Lesson extract at different concentrations, and the culture medium containing the drugs was replaced every 24 h. And then, at 72 hpf, the larvalmorphology and intersegmental vessels were observed under a microscope. The hatchability of 48-and 72-hpf embryos, and the number of intersegmen-tal vessels and the heart rate of 72-hpf juveniles were measured. Results:Compared with the control group, when the concentration of Whitmania pigra Whitmanis was higher than 30μg· ml-1 , and the concentration of Hirudinaria manillensis Lesson was higher than 20μg· ml-1, the number of intersegmental vessels was significantly reduced (P<0. 01). Compared with the control group, at 48 hpf, when the concentration of the drug groups was higher than 40 μg· ml-1 , the hatchability of the two groups significantly decreased ( P<0. 01);at 72 hpf, the hatchability of Whitmania pigra Whitman decreased significantly at the concentration of 100 μg·ml-1 (P<0.01), while the hatchability of Hirudinaria manillensis Lesson decreased significantly at the concentration of 80 μg· ml-1(P <0. 01). There was no obvious yolk sac edema, pericardial edema and spine curvature in the two groups. The heart rate decreased sig-nificantly (P<0. 01), while was still within the normal range. Conclusion:Both Whitmania pigra Whitman and Hirudinaria manillen-sis Lesson have notable anti-angiogenic activity, and the anti-angiogenesis activity of Hirudinaria manillensis Lesson is stronger. They both have effects on the development of zebrafish embryos, while the toxicity is not obvious.

OBJECTIVETo detect potential mutation of TCOF1 gene in a Chinese family affected with Treacher-Collins syndrome.

METHODSClinical data of the patient was collected. The analysis included history taking, clinical examination and genetic testing. All coding regions of the TCOF1 gene were subjected to PCR amplification and Sanger sequencing.

RESULTSA novel mutation c.2261ins G (p.E95X) of the TCOF1 gene was discovered in the patient. The same mutation was not found in his parents and 100 healthy controls.

CONCLUSIONThe c.2261insG (p.E95X) mutation of the TCOF1 gene probably underlies the disease in the patient. Genetic testing can facilitate diagnosis and genetic counseling for families affected with TCS.

Objective: To explore the effects of BPA on the expression of N-cadherin, Vimentin and FSHR in rat Sertoli cells. Methods: Primary Sertoli cells collected from prepuberty rats (18-21 d) were cultured for 48 h, and then they were treated with 0, 30, 50, 70 μmol/L BPA respectively for 24 h. The methods of MTT, real-time quantitative PCR and Western blotting were utilized to measure the cell ability of Sertoli cells, the mRNA and protein expression levels of N-cadherin, Vimentin and FSHR respectively. Results: Compared with control, the cell abilities of Sertoli cells in 50 μmol/L BPA group and 70 μmol/L BPA group increased significantly (P<0.05) . The cell abilities of Sertoli cells decreased with the increases of exposure doses of BPA. Compared with control, the expression of N-cadherin mRNA only increased in 30 μmol/L BPA group (P<0.05) , the expression of Vimentin mRNA decreased significantly in all doses group of BPA (P<0.05) , the expression of FSHR mRNA increased in all doses group of BPA (P<0.05) . Compared with the control, the protein levels of N-cadherin increased significantly in 50 μmol/L BPA group (P<0.05) , the protein levels of Vimentin decreased significantly in all doses group of BPA (P<0.05) , the protein levels of FSHR decreased significantly in 50 μmol/L BPA group and 70 μmol/L BPA group (P<0.05) . Conclusion The mechanism of testicular toxicity from BPA might be the alterations of N-cadherin, Vimentin and FSHR by disturbing normal spermatogenesis.

【Objective】 To revise the Social Connectedness Scale-Revised （SCS-R） so as to explore the factor structure of SCS-R in the context of Chinese culture and test its reliability and validity. 【Methods】 Totally 900 college students were asked to complete the Chinese version of the Social Connectedness， and 850 valid questionnaires were collected. In addition， the Distress Disclosure scale （DDI） and UCLA Loneliness Scale were used as the criteria. One month later， 100 students were randomly selected for retest. 【Results】 ① The results of exploratory factor analysis showed that one factor with more than one eigenvalue was selected， and the cumulative variance explained was 62.44%. ② Confirmatory factor analysis showed that the data fit well （χ²=33.438， df=14， χ²/df=2.388， RMSEA=0.04， CFI=0.996， TLI=0.991）. ③ The Chinese version of the social connectedness scale is a one-dimensional scale. Its internal consistency coefficient is 0.916 and test-retest reliability is 0.845. ④ Social connectedness is significantly positively correlated with distress disclosure and negatively correlated with loneliness， indicating that the scale has good criterion-related validity. 【Conclusion】 The Chinese version of the social connectedness scale has satisfactory reliability and validity， and has cross-cultural adaptability.