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Objective:To establish Sprague-Dawley (SD) rat models of cognitive impairment through repeated stimulation of lipopolysaccharide (LPS) in the early brain development, and to inquire into the effect of " multi-hits" mediated by inflammatory response on the histology and behavior of SD rat models and related molecular mechanisms.Methods:This study adopted a group design for experiments.The " multi-hits" SD rat models were established by intraperitoneal injection of LPS.According to the random number table method, 24 pregnant rats were randomly divided into 4 groups: control group, LPS1 group, LPS2 group and LPS3 group, 6 rats in each group.In the control group, saline was intraperitoneally injected into rats with gestational age of 18 days and 20-day-old neonatal rats.Rats with gestational age of 18 days were intraperitoneally injected with saline in the LPS1 group, 0.05 mg/kg LPS in the LPS2 group, and 0.1 mg/kg LPS in the LPS3 group.The pups in LPS1-3 groups were all injected intraperitoneally with 1 mg/kg LPS at the postnatal age of 20 days.The motor and cognitive function of the pups were evaluated overall by behavioral experiments such as forelimb suspension tests, grid tests and water maze tests.The relative expression of glial fibrillary acidic protein (GFAP), Notch1 and Jagged1 in brain tissue of pups was mainly detected by Western blot (WB) and histological experiments.One-way ANOVA analysis of variance and independent samples t- test were used to compare data among groups and between groups, respectively. Results:(1) Behavioral experiments: compared with the control group, LPS1-3 groups showed progressive decrease in forelimb suspension time [(34.81±5.66) s, (22.47±4.35) s, and (13.20±4.25) s vs.(43.88 ± 4.85) s], and the number of missteps in the grid experiment increased progressively (16.13±2.90, 20.75±3.10, 25.13±4.45 vs.9.00±2.72). The differences were statistically significant ( F=69.77, 35.59, all P<0.001). Both the escape latency and total distance in Morri′s water maze test increased progressively ( P<0.05). (2) WB experiment: the relative expression levels of GFAP, Notch1 and Jagged1 proteins in LPS1-3 groups were significantly higher than that in the control group ( P<0.05). (3) Hematoxylin-eosin (HE) staining and electron microscope pathology: compared with the control group, LPS1-3 groups had more loosely arranged frontal cortices and more obvious cell pyknosis.Under the electron microscope, the cytoplasm was swelling to varying degrees, mitochondrial cristae were broken, and part of the nuclear membrane was damaged. Conclusions:In the " multi-hits" cognitive impairment model, the damage to the brain tissue structure and behavioral changes of pups may be related to the up-regulation of Notch1/Jagged1 pathway mediated by repeated exposure to LPS.
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Since its establishment 30 years ago, the discipline of metabolic engineering has developed rapidly based on its deep integration with molecular biology, systems biology and synthetic biology successively, which has greatly contributed to advancing and upgrading biotechnology industry. This review firstly analyzes the current status of academic research and China's competence in the area of metabolic engineering according to the data of papers published in SCI-indexed journals in the past 30 years. Subsequently, the article summarizes the development of systems biology methods and enabling technologies of synthetic biology and their applications in metabolic engineering in the past 10 years. Finally, the major challenges and future perspectives for the development of metabolic engineering are briefly discussed.
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Biotechnology , Industry , Metabolic Engineering , Synthetic Biology , Systems BiologyABSTRACT
Objective@#To compare the efficacy of goal-activity-motor environment (GAME) therapy and neurodevelopmental therapy (NDT) in the early intervention of high-risk infants with cerebral palsy (IHRCP), and to provide scientific evidence-based medical basis for early intervention of IHRCP.@*Methods@#A total of 62 cases of IHRCP were enrolled in the Children′s Neurological Rehabilitation Center of the First Affiliated Hospital of Anhui Medi-cal University from June 2017 to December 2018.They were divided into GAME group (32 cases) and NDT group (30 cases) according to the admission order.Gross Motor Function Scale (GMFM), Fine Motor Function Measure (FMFM) and Gesell Development Scale (GDS) were used for detection and comparison.The differences among the gross motor, the fine motor score and the developmental quotient (DQ) between two groups before treatment, 9 months after treatment and 12 months after treatment, and the normalization rate and the incidence of cerebral palsy between the two groups at 12 months of age were compared.@*Results@#(1) Motor function was as follows: at 9 months[GAME: (32.63±15.83) scores, (30.03±15.88) scores], [NDT: (33.37±15.61) scores, (29.67±12.54) scores] and at 12 months[GAME: (40.56±15.79) scores, (36.31±14.98) scores], [NDT: (40.47±15.50) scores, (36.73±14.58) scores] after treatment, and GMFM and FMFM scores in GAME and NDT groups were significantly higher than those before treatment[GAME: (27.56±14.24) scores, (21.75±11.35) scores], [NDT: (26.93±14.96) scores, (21.30±10.67) scores], and the differences were significant (all P<0.01), but there was no significant difference between the 2 groups (all P> 0.05). (2) DQ had no significant difference in DQ between GAME group(63.59±10.83) and NDT group (61.59±7.96) before treatment (P>0.05). The total DQ at 9 months, 12 months, the total DQ of GAME group (73.67±12.00, 81.59±13.03) was significantly higher than that of NDT group (66.05±9.54, 75.17±1.92) (all P<0.05). Among them, the improvement of GAME in speech (79.84±16.56, 83.19±17.05) at 9 months and 12 months, and adaptive ability(78.63±16.37, 85.78±13.60) were significantly higher than that of NDT group(71.63±13.36, 72.53±12.77), (68.20±14.97, 77.43±12.10), and the differences were significant (all P<0.05). (3) Prognosis was as follows: at 12 months after treatment, 25 cases in GAME group and 23 cases in NDT group developed into normal children, there was no significant difference in the normalization rate between the 2 groups (P>0.05); the incidence of cerebral palsy was present in 6 cases in GAME group and 5 cases in NDT group, and there was no significant difference between the 2 groups (P>0.05).@*Conclusions@#GAME therapy and NDT had significant effects on both gross and fine exercise of IHRCP, and the efficacy of the two methods is similar.Both GAME therapy and NDT can equally promote IHRCP development into normal infants and reduce the incidence of cerebral palsy.
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Objective To compare the efficacy of goal_activity_motor environment( xAmE)therapy and neurodevelopmental therapy(NDT)in the early intervention of high_risk infants with cerebral palsy(IHRCP),and to provide scientific evidence_based medical basis for early intervention of IHRCP. Methods A total of 62 cases of IHRCP were enrolled in the Children＇s Neurological Rehabilitation Center of the First Affiliated Hospital of Anhui medi_cal University from June 2017 to December 2018. They were divided into xAmE group(32 cases)and NDT group(30 cases)according to the admission order. xross motor Function Scale(xmFm),Fine motor Function measure(FmFm) and xesell Development Scale(xDS)were used for detection and comparison. The differences among the gross motor, the fine motor score and the developmental quotient( D匝)between two groups before treatment,9 months after treat_ment and 12 months after treatment,and the normalization rate and the incidence of cerebral palsy between the two groups at 12 months of age were compared. Results ( 1 )motor function was as follows:at 9 months[ xAmE:(32. 63 ± 15. 83)scores,(30. 03 ± 15. 88)scores],[NDT:(33. 37 ± 15. 61)scores,(29. 67 ± 12. 54)scores]and at 12 months[xAmE:(40. 56 ± 15. 79)scores,(36. 31 ± 14. 98)scores],[NDT:(40. 47 ± 15. 50)scores,(36. 73 ± 14. 58)scores]after treatment,and xmFm and FmFm scores in xAmE and NDT groups were significantly higher than those before treatment[xAmE:(27. 56 ± 14. 24)scores,(21. 75 ± 11. 35)scores],[ NDT:(26. 93 ± 14. 96)scores, (21. 30 ± 10. 67)scores],and the differences were significant( all P<0. 01),but there was no significant difference between the 2 groups(all P> 0. 05).(2)D匝 had no significant difference in D匝 between xAmE group(63. 59 ± 10. 83)and NDT group(61. 59 ± 7. 96)before treatment(P>0. 05). The total D匝 at 9 months,12 months,the total D匝 of xAmE group(73. 67 ± 12. 00,81. 59 ± 13. 03)was significantly higher than that of NDT group(66. 05 ± 9. 54, 75. 17 ± 1. 92)(all P<0. 05). Among them,the improvement of xAmE in speech(79. 84 ± 16. 56,83. 19 ± 17. 05)at 9 months and 12 months,and adaptive ability(78. 63 ± 16. 37,85. 78 ± 13. 60)were significantly higher than that of NDT group(71. 63 ± 13. 36,72. 53 ± 12. 77),(68. 20 ± 14. 97,77. 43 ± 12. 10),and the differences were significant( all P<0. 05).(3)Prognosis was as follows:at 12 months after treatment,25 cases in xAmE group and 23 cases in NDT group developed into normal children,there was no significant difference in the normalization rate between the 2 groups( P>0. 05);the incidence of cerebral palsy was present in 6 cases in xAmE group and 5 cases in NDT group,and there was no significant difference between the 2 groups(P>0. 05). Conclusions xAmE therapy and NDT had significant effects on both gross and fine exercise of IHRCP,and the efficacy of the two methods is similar. Both xAmE therapy and NDT can equally promote IHRCP development into normal infants and reduce the incidence of cerebral palsy.
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Objective To analyze and identify the protein of patients with IgA nephropathy for identification and quantitative a-nalysis using the combination of liquid chromatography and mass spectrometry which combined isobaric tags for relative and abso-lute quantification .Methods Proteomic analysis of renal tissue in patients with IgA nephropathy and normal controls was per-formed to identify different expressed proteins .Results A total of 1860 proteins were identified ,287 proteins were upregulated in renal tissue of IgA nephropathy patients ,287 proteins were downregulated significantly ,finally 9 upregulated proteins and 8 down regulated proteins were identified(protein fold difference greater than 1 .5) .Conclusion Quantitative proteomic technology is effi-ciently applicable for identification and relative quantitation of proteome in renal tissue ,which could get all the information of differ-ent protein .This is useful for us to better understand the relationship of protein and the pathogenesis of IGA nephropathy .
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Objective To analyze and identify the protein of patients with IgA nephropathy for identification and quantitative a-nalysis using the combination of liquid chromatography and mass spectrometry which combined isobaric tags for relative and abso-lute quantification .Methods Proteomic analysis of renal tissue in patients with IgA nephropathy and normal controls was per-formed to identify different expressed proteins .Results A total of 1860 proteins were identified ,287 proteins were upregulated in renal tissue of IgA nephropathy patients ,287 proteins were downregulated significantly ,finally 9 upregulated proteins and 8 down regulated proteins were identified(protein fold difference greater than 1 .5) .Conclusion Quantitative proteomic technology is effi-ciently applicable for identification and relative quantitation of proteome in renal tissue ,which could get all the information of differ-ent protein .This is useful for us to better understand the relationship of protein and the pathogenesis of IGA nephropathy .