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Objective:To investigate the neonatal outcomes and prognosis of fetuses who were prenatally diagnosed with intraventricular hemorrhage(IVH) to provide evidence for clinical consultation and management.Methods:Clinical data of fetal IVH cases ( n=89) diagnosed by fetal neurosonogram (NSG) in Peking University First Hospital from January 2012 to April 2020 were retrospectively collected, and neonatal outcomes were analyzed, involving fetuses with different grades of IVH and coexisting abnormalities. These patients were followed up for more than three months after birth. Two child development screening systems, Ages & Stages Questionnaires (Third Edition) (ASQ-3) and Ages & Stages Questionnaires: Social-Emotional (ASQ-SE), were used to assess the development of the patients from several aspects including physical growth, oral communication, motor ability and social emotions. Descriptive statistical analysis was used in this study. Results:(1) A total of 89 fetuses were enrolled and 66.3% (59/89) of them underwent fetal cranial MRI examination after ultrasound diagnosis. Among these 59 cases, 32(54.2%) had the same results with fetal NSG; 20(33.9%) with the diagnosis of remote ventricular cystic hemorrhage by fetal NSG, but misdiagnosed by MRI, were all confirmed by neonatal craniocerebral ultrasound; 7(11.9%) were lost to follow-up or terminated. In addition to IVH that was consistent with the ultrasound diagnosis, MRI also found three cases of cortical malformation, three cases of subdural hemorrhage and two cases of cerebral parenchymal hemorrhage. (2) Among the 89 cases, 37 (41.6%) underwent neonatal craniocerebral ultrasound examination after birth showing small amount of remote IVH, which was consistent with previous fetal NSG diagnosis. (3) There were 38 cases complicated by lateral ventricular widening (three lost to follow-up, 18 were terminated , one died in uterus, 15 with good and one with poor outcome), 10 cases by severe IVH sequelae (one lost to follow-up and nine were terminated) and 10 by craniocerebral malformation (one lost to follow-up, eight were terminated and one with poor neonatal prognosis). Two cases with extracranial malformation (ureteropelvic junction obstruction) were healthy after birth. (4) There were 29, 30, 16 and 14 cases of gradeⅠ,Ⅱ,Ⅲ andⅣ of fetal IVH, respectively. Among all cases, 12 were lost to follow-up; three died in uterus (all with gradeⅣ IVH); 31 were terminated and 43 were born and followed up. During the follow-up of the 43 cases, two (one grade Ⅰ case with corpus callosum dysplasia and one grade Ⅲ case with fetal cytomegalovirus infection) had poor prognosis, while the other 41 (one gradeⅢ, 17 gradeⅡ and 23 gradeⅠcases) showed no severe abnormalities.Conclusions:Fetal NSG is the first choice in screening fetal IVH and MRI can be a valuable aid. The neonatal prognosis may be associated with the grade of hemorrhage and coexisting abnormalities. Fetuses with isolated grade Ⅰ or Ⅱ IVH usually have a relatively good prognosis.
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Objective:To construct the pET30a-EgG1Y162-2 prokaryotic expression plasmid and induce the expression of EgG1Y162-2 protein, so as to provide a research basis for development of Echinococcus granulosus vaccine. Methods:Using Echinococcus granulosus cDNA as a template, the target gene of EgG1Y162-2 was synthesized by PCR, and after digestion with restriction enzymes EcoRⅠ and Hind Ⅲ, it was connected to the prokaryotic expression vector pET30a to construct the recombinant plasmid pET30a-EgG1Y162-2. The recombinant plasmid was transformed into competent cell BL21 (DE3) and induced by isopropyl β-D-thiogalactoside (IPTG) to express a large number of proteins. The recombinant protein was purified by affinity chromatography. The purification level was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and the expression product was identified by Western blotting. Results:The recombinant plasmid pET30a-EgG1Y162-2 was successfully constructed. After inducting expression, the bacterial supernatant and the eluate were both at a relative molecular weight of about 15 × 10 3, and the protein antigen component eluted with 200 mmol/L imidazole was relatively pure. Western blotting results showed that the purified recombinant protein EgG1Y162-2 with His tag could be recognized by His monoclonal antibody. Conclusion:The pET30a-EgG1Y162-2 prokaryotic expression plasmid of Echinococcus granulosus is successfully constructed, and the recombinant protein of EgG1Y162-2 is induced to express, laying a foundation for further study on anti- Echinococcus granulosus vaccine.
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This study aimed to explore the link between block copolymers' interfacial properties and nanoscale carrier formation and found out the influence of length ratio on these characters to optimize drug delivery system. A library of diblock copolymers of PEG-PCL and triblock copolymers with additional PEI (PEG-PCL-PEI) were synthesized. Subsequently, a systematic isothermal investigation was performed to explore molecular arrangements of copolymers at air/water interface. Then, structural properties and drug encapsulation in self-assembly were investigated with DLS, SLS and TEM. We found the additional hydrogen bond in the PEG-PCL-PEI contributes to film stability upon the hydrophobic interaction compared with PEG-PCL. PEG-PCL-PEI assemble into smaller micelle-like (such as PEG-PCL4006-PEI) or particle-like structure (such as PEG-PCL8636-PEI) determined by their hydrophilic and hydrophobic block ratio. The distinct structural architectures of copolymer are consistent between interface and self-assembly. Despite the disparity of constituent ratio, we discovered the arrangement of both chains guarantees balanced hydrophilic-hydrophobic ratio in self-assembly to form stable construction. Meanwhile, the structural differences were found to have significant influence on model drugs incorporation including docetaxel and siRNA. Taken together, these findings indicate the correlation between molecular arrangement and self-assembly and inspire us to tune block compositions to achieve desired nanostructure and drug loading.
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Objective:To explore the clinical efficacy and safety of the new targeted anti-immunoglobulin E (IgE) drug Omalizumab in the treatment of children with moderate and severe allergic bronchial asthma in China.Methods:The clinical data of children with moderate and severe allergic asthma who were treated in the Department of Pediatrics of Peking University First Hospital from July 2018 to January 2020 and treated with Omalizumab were retrospectively analyzed.A comparison was performed on the changes of questionnaires, including childhood asthma control test (C-ACT), pediatric asthma quality of life questionnaire (PAQLQ), mini-asthma quality of life questionnaire (Mini-AQLQ) and global evaluation of asthma treatment effectiveness (GETE), pulmonary function test, including forced expiratory volume in the first second (FEV 1), percentage to predicted value (FEV 1% pred) and small airway function, fractional concentration of exhaled nitric oxide (FeNO) and so on, before and after treating with Omalizumab. Results:A total of 15 pediatric patients completed 16 weeks of treatment follow-up.After 16 weeks of Omazumab treatment, the score of C-ACT increased from (16.001.66) scores to (25.38±0.64) scores ( F=11.969, P<0.001), PAQLQ increased from (118.08 ± 23.78) scores to (141.00 ± 11.91) scores ( F=9.289, P=0.001), Mini-AQLQ increased from (78.93±7.43) scores to (97.92±3.12) scores ( F=4.145, P=0.042), and GETE decreased from (2.47±0.27) scores to (1.60±0.19) scores ( t=2.982, P=0.010). The actual value of FEV 1, FEV 1% pred, the maximum expiratory flow percentage of the predicted value (PEF% pred), forced expiratory flow at 25% (MEF 75), forced expiratory flow at 50% (MEF 50) and maximum midexpiratory flow (MMEF 75/25), increased from (1.96±0.12) L to (2.17±0.21) L ( F=0.425, P=0.789), (81.46±2.85)% to (82.64±1.55)% ( F=0.926, P=0.465), (82.05±3.58)% to (91.10±4.67)% ( F=1.909, P=0.128), (60.36±7.43)% to (76.94±4.65)% ( F=2.120, P=0.096), (52.72±3.75)% to (73.80±8.34)% ( F=3.140, P=0.047) and (60.05±8.47)% to (74.86±10.85)% ( F=7.860, P=0.010), respectively.FeNO decreased from 25.0 (14.5, 35.5) μg/L to 20.0 (18.5, 30.0) μg/L ( Z=-0.206, P=0.840). Transient headache was observed in 1 case and evanescent eruption in 2 cases during the treatment, respectively. Conclusions:Omalizumab can significantly improve the clinical manifestations, lung function indicators and quality of life of children with moderate and severe allergic asthma, and has good safety.Thus, it is expected to play an important role in the treatment of children with moderate-to-severe allergic asthma.
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OBJECTIVE@#To establish a stable HEK293T cell line with c.392G>T (p.131G>V) mutation site knockout in gene using CRISPR/Cas9 technique.@*METHODS@#We designed 4 pairs of small guide RNA (sgRNA) for c.392G>T(p.131G>V) mutation site, and constructed exogenous PX458 plasmids expressing Cas9-sgRNA. The plasmids were transfected into HEK293T cells, and the cells expressing GFP fluorescent protein were separated by flow cytometry for further culture. After verification of the knockout efficiency using T7 endonuclease Ⅰ, the monoclonal cells were screened by limiting dilution and DNA sequencing to confirm the knockout. We detected the expressions of mRNA and protein and examined functional changes of the genetically modified cells.@*RESULTS@#We successfully constructed the Cas9-sgRNA exogenous PX458 plasmid based on the c.392G>T(p.131G>V) mutation site of gene. The editing efficiency of the 4 pairs of sgRNA, as detected by T7E1 enzyme digestion, was 6.74%, 12.36%, 12.54% and 2.94%. Sanger sequencing confirmed that the HEK293T cell line with stable knockout of c.392G>T(p.131G>V) was successfully constructed. The genetically modified cells expressed lower levels of mRNA and protein and showed reduced enzyme activity and proliferative capacity and increased apoptosis in response to vitamin K3 treatment.@*CONCLUSIONS@#We successfully constructed a stable HEK293T cell model with gene c.392G>T(p.131G>V) mutation site knockout to facilitate future study of gene repair.
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Humans , CRISPR-Cas Systems , HEK293 Cells , Mutation , PlasmidsABSTRACT
Objective To study the neurological prognosis of neonates with ventriculomegaly and its influencing factors.Method A retrospective study was conducted among neonates with ventriculomegaly in Peking University First Hospital from January 2013 to December 2015.A series of cranial ultrasonography were performed after birth and the Gesell development scale was conpleted after six months.x2 test and two-independent-sample t test were used for statistical analysis.Result Among 103 cases of ventriculomegaly,95 cases (92.2％) had mildly enlarged lateral ventricles and 8 cases (7.8％) significantly enlarged.83 cases received serial cranial ultrasound examinations after birth.The lateral ventricles of 9 patients (10.8％) bacame wider and 74 (89.2％) not.The Gesell development scales were completed in 65 cases 6 months after birth.Among them,8 patients with widening lateral ventricles got poor prognosis (100％).Among 57 patients without progressively widening lateral ventricles,6 (10.5％) had poor prognosis.The difference was statistically significant (P ＜ 0.001).No correlation was found between the severity of the lateral ventricle widening and the neurological outcome (P =1.000).There were 2 cases with other abnormalities,and 1 case got poor prognosis on follow-up.Conclusion Most neonatal ventriculomegaly patients have mild and isolated lateral ventricle enlargement.Most of them remain stable or gradually return to normal.The patients with progressively widening lateral ventricles are likely to have adverse neurological prognosis.
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Objective To explore the prevalence and risk factors of stroke in a population over 40 years in a community in Shenzhen, China. Methods The subjects investigated were a population ≥40 years old in this cross-sectional study. Cluster sampling method was used to conduct the unified questionnaires, physical examination, and laboratory examination in the community residents, and then the survey data were used for online entry analysis. Results A total of 5 308 community residents were screened, and 160 experienced stroke. The crude prevalence of stroke was 3 014. 32/100 000. The prevalence of stroke increased with age, and that in males was significantly higher than that in females (3 721. 37/100 000 vs. 2 552. 93/100 000; χ2 = 5. 923, P = 0. 015). There were significant differences in the proportions of males, obvious overweight, hypertension, diabetes mellitus, atrial fibrillation, lack of physical activity, and family history of stroke, as well as age, systolic blood pressure, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, and fasting glucose level between the stroke population and the non-stroke population (all P 80 years) in both males and females. The proportions of diabetes (P < 0. 001) and atrial fibrillation (P < 0. 001) in males, and obvious overweight (P = 0. 001) in females were gradually increased with age, and the proportion of smoking in males weas gradually reduced with age. The proportions of hypertension (P < 0. 001) and obvious overweight (P < 0. 001) in males were significantly higher than those in females at the age of 40 to 49 years. The proportions of hypertension (P < 0. 001), diabetes (P < 0. 001) and obvious overweight (P < 0. 001) in males were significantly higher than those in females at the age of 50 to 59 years. The proportion of hypertension in males was significantly higher than that in females at the age of 60 to 69 years (P = 0. 039). The proportions of hypertension (P = 0. 016), atrial fibrillation (P = 0. 028) and hyperlipidemia (P = 0. 023) in females were significantly higher than those in males at the age of 70 to 79 years. The proportion of obvious overweight in females was significantly higher than that in males at the age of ≥80 years (P =0. 001). Conclusions The crude prevalence of stroke is higher among the community residents. The the levels of exposure to stroke risk factors including hypertension, diabetes and atrial fibrillation are higher. It may be important to intervene on these risk factors in community residents, especially in elders and those with family history of stroke.
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Objective To analyze the changes of electroencephalographic activities during fully automated simultaneous peripheral arteriovenous exchange transfusion(ET) in neonates with hyperbilirubinemia.Methods A total of 45 neonates who suffered from severe hyperbilirubinemia and underwent fully automated simultaneous peripheral arteriovenous exchange transfusion were studied from March 2009 to March 2016 in Neonatal Intensive Care Unit of Guangzhou Women and Children's Medical Center,and 46 ETs were performed in 45 babies who were divided into 2 groups according to the severity of hyperbilirubinemia:the encephalopathy group and the none-encephalopathy group.Nineteen patients were in the encephalopathy group,in which 11 were male and 8 were female.The other 26 patients were in the none-encephalopathy group,in which 15 were male and 11 were female.Changes in amplitude integrated electroencephalogram(aEEG) during ETs were analyzed,including background activities,sleep-wake cycle (SWC)and seizures.Results Forty-five patients with hyperbilirubinemia underwent 46 fully automated simultaneous peripheral arteriovenous ETs.As a result,total bilirubin dropped from (524.90 ± 110.96)μmol/L before ETs to (245.62 ±78.97) μmol/L after ETs,with clearance rate of 53.2％.And indirect bilirubin dropped from(486.16 ±90.39) μmol/L before ETs to(222.19 ± 79.49) μmoL/L after ETs,with clearance rate of 54.3％.On the other hand,there was no significant difference in the changes of electroencephalographic activities during ETs,including background activities (x2 =0.16,P ＞ 0.05),SWC (x2 =0.71,P ＞ 0.05) and seizures (x2 =0.30,P ＞ 0.05).However,there were significant difference in suppressions on background activities between the encephalopathy group and the none-encephalopathy group(Fisher's exact test P =0.042),though there were no significant statistical differences in SWC or seizures between the 2 groups (x2 =0.65,P ＞ 0.05;x2 =2.07,P ＞ 0.05,respectively).Conclusions In neonatal hyperbilirubinemia,fully automated simultaneous peripheral arteriovenous ET is safe and efficient without significant influence on electroencephalographic activities as a whole.However,background activities are more significantly depressed in infants of bilirubin encephalopathy than that of non-encephalopathy during ET.
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Objective To study the characteristics and its risk fastors of brain development of the preterm infant early after birth in amplitude-integrated electroencephalography(aEEG). Methods The 153 preterm infants who had seen a doctor in Peking University First Hospital from April 2009 to August 2013 accepted the aEEG check at term of corrected gestational age ( ≥ 38 weeks but < 42 weeks of corrected gestational age). The risk factors of brain development, such as gestational age ( < 30, 30 ≤ - ≤ 33+6 and 34≤-≤36+6 weeks), clinical informations [relatively stable group including 104 cases without any serious complications or brain injury, the group only suffering from a serious brain injury (19 cases), and the group only suffering from severe systemic disease (30 cases)] and nutrition (good or malnutrition), were analyzed. Also the relationship between the aEEG and the cranial ultrasound detected at the same time and the Gesell Developmental Scale at six months of corrected gestational age. Theχ2 test, two independent samples t-test and Logistic regression analysis were used for statistical analysis. Results The aEEG of 52%(79/153) cases reached the level of normal full-term newborn at term of corrected gestational age, only 48% (74/153) were abnormal. The abnormal rate of aEEG results in relatively stable preterm infants decreased from 3/6 (<30 weeks) to 35%(13/37) at 34 ≤ - ≤ 36+6 weeks, but the difference was not statistically significant (χ2=1.998, P=0.353). The abnormal rate of aEEG results in the group suffering from a serious brain injury was higher than the relatively stable preterm infants [14/19 vs 44%(46/104) ,χ2=5.578, P=0.024]. In relatively stable preterm infants, there was no difference of the abnormal rate of the aEEG results between intrauterine malnutrition group and good nutrition group [46%(19/41) vs 43%(27/63),χ2=0.122, P=0.727]. Neither was between extrauterine malnutrition group and good nutrition group [52%(13/25) vs 42%(33/79),χ2=0.805, P=0.369]. Serious brain injury was independent risk factor of abnormal aEEG (OR=3.453, 95%CI: 1.177-10.132, P=0.024). The coincidence rate of aEEG and the cranial ultrasound examination or the scores of Gesell Developmental Scale was 57%(56/98) and 50%(10/20), respectively. Conclusions The brain catch-up development may appears early after birth in preterm infants, which are impaired by lower gestational age and the severe brain injury. It is more effective of aEEG for evaluating the brain development of preterm infants when combines with other methods.
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Objective To study clinical efficacy on cesarean scar pregnancy (CSP) treated by transvaginal surgery.Methods From Jan.2008 to Mar.2011,31 cases with CSP were managed by transvaginal surgery in Anshan Women and Children Hospital.Based on ultrasonograpy examination and intraoperative exposure of lesion,variable surgical options were executed.Fifteen cases in group A were treated by debridement resection and vaginal repair of uterine wall,7 cases in group B were treated by transvaginal uterine artery ligation and curettage,9 cases were treated by cutting the anterior wall in the lower uterine segment and repairing uterine.The intraoperative blood loss,operation time,hospital stay,hCG fluctuation at postoperative period and complications were analyzed among those groups.Results Allcases in 3 groups were cured well in one time.( 1 ) The intraoperative blood loss were (41 ±21 ) ml in group A,(27 ±7) ml in group B and ( 148 ± 132) ml in group C.There was no statistically different blood loss between group A and group B ( P ＞ 0.05 ),however,the amount blood loss in group C was significantly more than those in group A and group B ( P ＜ 0.05 ).( 2 ) The average surgical time,the mean hospital stay,postoperative recovery time of blood hCG were (40 ± 11 ) minutes,(4.7 ± 0.8 ) days and ( 2.7 ± 1.0) weeks in group A,(44 ± 5 ) minutes,(4.0 ± 0.8) days and (2.9 ± 1.0) weeks in group B,(40 ± 12) minutes,(4.9 ± 1.0) days and (2.8 ±0.9) weeks in group C.Those clinical index were no statistically different among those 3 groups(P ＞0.05).(3) No bladder injury and other complications were observed in those groups.Conclusions Transvaginal surgery is efficacy,easy to operate,to keep the uterus,safe and economy in treatment of CSP.Surgery in group A is suitable to treat early and exogenous lesions; surgery in group B is suitable to treat endogenous lesions; surgery in group C is suitable to treat failure cases in group A and B,however,the injury is greater than those in group A and B.
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Objective:To compare the newborn piglet models of hypoxic ischemic brain damage in hypoxia and hypoxia combined with occlusion of both carotid arteries. Methods: Twenty four 7-day-old piglets were divided into two groups. Group H: mechanical ventilation with low concentration of oxygen,Group HI; mechanical ventilation with low concentration of oxygen combined with occlusion of both carotid arteries. The piglets were inhaled with 10%, 8%, and 6% low-concentration oxygen for 30 min, and grouped into mild, moderate, and severe hypoxia ones. The changes of physiological parameter, cerebral blood flow and cerebral oxygen perfusion were detected. Results: There were no significant differences in blood gas analysis of oxygen saturation, blood lactic acid and pH between the two groups(P＞0. 05) . The mean arterial pressure of severe hypoxia in HI was significantly lower than in H(P＜0.05). The cerebral blood flow in H and HI was relatively stable after different degrees of hypoxia. As compared with the cerebral blood flow perfusion in group H and HI, there were no significant differences between them ( P>0. 05). The cerebral oxygen perfusion in H and HI was significantly descent after different degrees of hypoxia (P＜ 0.05). As compared with the cerebral oxygen perfusion in groups H and HI, there were no significant differences between them. Conclusion: H and HI have the same effect on physiological parameter, cerebral volume and cerebral oxygen perfusion of newborn piglets. The mechanical ventilation with low concentration of oxygen to newborn piglets can develop the HIBD model, it is not necessary to occlude carotid arteries.
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The neonatal burst suppression is a severe EEG pattern and always demonstrates serious damage of nerve system. But the outcome of these patients depends on the different etiology. A total of 256 cases of video EEG recordings were analyzed in order to summarize the etiology and outcome of burst suppression. The results showed that some patients in all 17 cases of burst suppression showed EEG improvement. The etiology was the dominant factor in long term outcome. It was suggested that effective video EEG monitoring is helpful for etiologic study and prognosis evaluation.
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Electroencephalography/methods , Hypoxia-Ischemia, Brain/diagnosis , Monitoring, Physiologic/methods , Neonatology/methods , Video RecordingABSTRACT
The neonatal burst suppression is a severe EEG pattern and always demonstrates serious damage of nerve system. But the outcome of these patients depends on the different etiology. A total of 256 cases of video EEG recordings were analyzed in order to summarize the etiology and outcome of burst suppression. The results showed that some patients in all 17 cases of burst suppression showed EEG improvement. The etiology was the dominant factor in long term outcome. It was sug-gested that effective video EEG monitoring is helpful for etiologic study and prognosis evaluation.
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OBJECTIVE To investigate the bacterial contamination status of ventilator systems and corresponding measurements.METHODS Bacterial culture samples of auxiliary equipments of 36 ventilators,including inlet and outlet tube,mask,humidifier,water collector,nebulizer,and air filter,were respectively collected.The culture results were reported to the infectious disease control office of the hospital.After improved the sterilization management,then repetition of bacterial culture was performed and the results were compared with the former.RESULTS The first cultural results demonstrated severe bacterial contamination and the positive rate of inlet tube,humidifer,water collector,nebulizer,and air filter was even higher than 50%.After countermeasures were conducted,the positive rate of second bacterial culture of all ventilator systems was significantly dropped to less than 5.5%.CONCLUSIONS Bacterial contamination of ventilator systems commonly exists which can be significantly diminished by improved conception of asepsis and standardized sterilization.
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Objective To explore the extraction technology of polysaccharides from Asterias amurensis and study their antibacterial activities in vitro. Methods Determined by spectroscopic methodology with phenol -sulfuric acid,the polysaccharide content was taken as inspecting index to select the extraction technology such as enzymatic hydrolysis,alkali,and alkali-enzymatic combined. Filter paper method was used to test the polysaccharides antibiotic activities in vitro. Results The optimum extraction technology was adding 2 times amount of 1 mol?L-1 sodium hydroxide,heating for 2h,under these conditions the greatest yield of starfish polysaccharides would be obtained which showed inhilitory effect on Staphylococcus aureus. Conclusion The alkal hydrolysis extraction technology is superior to the other methods; polysaccharides from Asterias amurensis showed antibiotic activity to some kinds of bacteria.