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Objective To study the effect of chemically synthesized Nogo-66 receptor ( NgR ) specific small interfering RNA ( siRNA) on nerve regeneration and function of newborn rats after hypoxic-ischemic brain damage ( HIBD) . Methods A total of 50 HIBD newborn rats were set up. They were randomly assigned OR allocated into siNgR group ( n=20 ) , normal saline ( NS ) group ( n=20 ) and HIBD group ( n=10 ) . The rats of siNgR group were given intraventricular injection of siNgR and transfection reagents ( 10μl ); the rats of NS group were given intraventricular injection of NS and transfection reagents (10μl);and the rats of HIBD group had no intervention. In addition to these three groups, there is another group, sham-operated group ( n=10 ) . The rats of sham-operated group were sham-operated ( common carotid artery was isolated but not ligated) and did not receive hypoxia-ischemia processing and intraventricular injection. Utilize water maze experiment to analyze the rats' escape time. The levels of NgR and GAP-43 protein in rats' brains were measured by immunohistochemistry and image analysis. Results RT-PCR gel electrophoresis results showed that the NgR cDNA stripe of siNgR group was not obvious, but the stripe of NS group was clear. At the same time, the GAPHD cDNA bands of the above two groups were both clear. There were more NgR positive immune reaction products ( brown particles) in NS group than in siNgR group. The number of GAP-43 positive cells by immunohistochemistry in sham-operated group, HIBD group, NS group and siNgR group was (33. 24±1. 32), (20. 14±1. 24), (18. 73±1. 41) and (28. 06±1. 78), respectively. The number of sham-operated group and siNgR group was greater than HIBD group and NS group ( P＜0. 05 ) . There was no statistical significant difference for the number of GAP-43 positive cells between sham-operated group and siNgR group ( P＞0. 05 ) . Water maze experiment results showed that the newborn rats ' average escape time ( s ) of HIBD group ( 58. 1 ± 10. 3,47. 2±10. 1, 42. 5±7. 6) was obviously longer than sham-operated group (34. 2±5. 6, 25. 7±6. 2, 21. 2±8. 1), and the difference was statistically significant (P＜0. 05). However, the average escape time of siNgR group (37. 5±9. 8, 29. 1±9. 8, 27. 2±9. 3) was obviously shorter than HIBD group and NS group (60. 7±5. 2, 49. 1±9. 9, 45. 3±9. 3), (P＜0. 05). Conclusions Chemically synthesized specific siRNA had the potential to interference the expression of NgR in the brain of newborn rats, and to a certain extent, could promote the nerve regeneration and neural functional recovery of rats.
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Objective By 1 H magnetic resonance spectroscopy( 1 H MRS) ,small for gestational age (SGA)and appropriate for gestational age(AGA) as the detection of brain metabolites and MRI plus soft-ware measurement in different brain areas of volume,investigate its cerebral metabolites and the changes of brain in different parts of the volume and significance. Methods Select 88 patients eligible infants, SGA group of 27 cases and AGA group of 21 cases of premature infants;SGA group of 22 cases and AGA group of 18 cases of term infants. Preterm infants with a gestational age of 32 to 36 weeks,term infants with a gesta-tional age of 37 to 41 weeks. Check time between 4 to 7 days old. Calculation of cerebrum volume,cerebellar volume and cerebrospinal fluid volume and intracranial volume,N-acetylaspartic acid(NAA),as 1H MRS area of metabolites measured right frontal choline compounds( Cho) and creatine compounds( Cr) wave,calcu-lation of Cho/Cr and NAA/Cho ratio of NAA/Cr. Results NAA/Cr,the cerebrum volume and intracranial volume of SGA in premature infants group,term infants group and mixed group were 0. 627 ± 0. 183,(2. 831 ±0. 199) ×105 mm3,(3. 178 ±0. 209) ×105 mm3;0. 706 ±0. 139,(3. 056 ±0. 217) ×105 mm3,(3. 411 ± 0. 212 ×105 mm3;0. 708 ± 0. 171,(2. 932 ± 0. 234) × 105 mm3,(3. 282 ± 0. 239) × 105 mm3,respective-ly. NAA/Cr,the cerebrum volume and intracranial volume of AGA in premature infants group,term infants group and mixed group were 0. 734 ± 0. 101,(2. 987 ± 0. 111) × 105 mm3,(3. 347 ± 0. 137) × 105 mm3;0. 805 ± 0. 106, ( 3. 228 ± 0. 284 ) × 105 mm3 , ( 3. 588 ± 0. 306 ) × 105 mm3; 0. 721 ± 0. 119, ( 3. 098 ± 0.240) ×105 mm3,(3.458 ±0.258) ×105 mm3,respectively. The data of SGA group were all lower than those in AGA group,which had significant difference(P0. 05,respectively). In the premature infants groups,the NAA/Cho of SGA group(0. 401 ± 0. 737) was lower than in the AGA group(0. 506 ± 0. 116), which had significant difference(P=0. 000). In the term infants groups,the NAA/Cho of SGA group(0. 483 ±0. 605) was lower than in the AGA group(0. 472 ± 0. 987),which had no significant difference(P =0. 653). In the AGA groups,NAA/Cr,NAA/Cho,cerebellar volume and cerebrospinal fluid volume of pre-mature infants group and term infants group had no significant difference ( P>0. 05 ) . Both of the cerebellar volume and cerebrospinal fluid volume between the premature infants AGA group and premature infants AGA group had no significant difference(P>0. 05). Conclusion Neurons in the brain,the cerebrum volume,the cranial cavity volume and NAA/Cr of SGA was significantly lower than those of AGA,but Cho/Cr,cerebel-lar volume and cerebrospinal fluid volume of SGA and AGA had no significant difference. NAA/Cr in the brain and the cerebrum volume of SGA may be associated with low volume of small nerve mental retarda-tion,worthy of further study.
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Objective To prepare the mB7-1-GPI-anchored Lewis vaccine and investigate its antitumor effects. Methods mB7-1-GPI was incorporated on Lewis tumor cells and mB7-1-GPI-anchoring tumor vaccine was prepared. The anti-tumor immunity induced by the prepared mB7-1-GPI-anchored Lewis tumor cell vaccine in tumor-bearing mice was observed. Results Flow cytometric analysis showed that mB7-1-GPI were positively expressed on the surface of Lewis tumor cells. After Lewis tumor cells incubated with mB7-1-GPI, the positive rate (PR) of mB7-1 antigen was 95.8% (0h), 93.6% (4h), 91.1% (8h) and the fluorescence intensity (FI) was 11.2(0h), 10. 6(4h), 9. 8(8h). The IL-2 and IFN-γ production of splenic lymphocytes + lewis cells was (25.9 ± 1.4) pg/ml, (56. 0± 3. 5 ) pg/ml. The IL-2 and IFN-γ production of splenic lymphocytes + lewis/mB7-1-GPI was ( 871.3 ± 10. 4 ) pg/ml, ( 1329. 0 ± 11.9 ) pg/ml. In 25 days, the mean diameter of tumor of Lewis/mB7-1 -GPI was shorter than Lewis( 1.4 ± 0. 21 )cm & ( 2. 5 ± 0. 27 )cm , P ＜ 0. 05 ). Lewis tumor cell-bearing C57BL/6 mice treated with Lewis/mB7-1-GPI vaccine survived much longer than mice treated with Lewis vaccine ( 75.2 ± 2. 0 ) d & (40. 2 ± 2. 0 ) d ( P ＜ 0. 05 ). Conclusion The Lewis tumor vaccine prepared with mB7-1-GPI fusion protein significantly inhibited the tumor growth in Lewis bearing mice. It represented an useful new strategy for attaching immunological factor onto tumor cell surfaces without genetic manipulation.