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<b>Objective</b> To investigate the differences and the immunocompatibility of wild-type (WT), four-gene modified (TKO/hCD55) and six-gene modified (TKO/hCD55/hCD46/hTBM) pig erythrocytes with human serum. <b>Methods</b> The blood samples were collected from 20 volunteers with different blood groups. WT, TKO/hCD55, TKO/hCD55/hCD46/hTBM pig erythrocytes, ABO-compatible (ABO-C) and ABO-incompatible (ABO-I) human erythrocytes were exposed to human serum of different blood groups, respectively. The blood agglutination and antigen-antibody binding levels (IgG, IgM) and complement-dependent cytotoxicity were detected. The immunocompatibility of two types of genetically modified pig erythrocytes with human serum was evaluated. <b>Results</b> No significant blood agglutination was observed in the ABO-C group. The blood agglutination levels in the WT and ABO-I groups were higher than those in the TKO/hCD55 and TKO/hCD55/hCD46/hTBM groups (all <i>P</i><0.001). The level of erythrocyte lysis in the WT group was higher than those in the ABO-C, TKO/hCD55 and TKO/hCD55/hCD46/hTBM groups. The level of erythrocyte lysis in the ABO-I group was higher than those in the TKO/hCD55 and TKO/hCD55/hCD46/hTBM groups (both <i>P</i><0.01). The pig erythrocyte binding level with IgM and IgG in the TKO/hCD55 group was lower than those in the WT and ABO-I groups. The pig erythrocyte binding level with IgG and IgM in the TKO/hCD55/hCD46/hTBM group was lower than that in the WT group and pig erythrocyte binding level with IgG was lower than that in the ABO-I group (all <i>P</i><0.05). <b>Conclusions</b> The immunocompatibility of genetically modified pig erythrocytes is better than that of wild-type pigs and close to that of ABO-C pigs. Humanized pig erythrocytes may be considered as a blood source when blood sources are extremely scarce.
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Objective:To investigate the protective effect of hypothermic antegrade machine perfusion against canine ischemic brain injury.Methods:Thirteen beagle dogs were divided into the mild hypothermia with perfusion group ( n=6) and normothermia with perfusion group ( n=7) according to the random number table. The model of ischemic brain injury was established by neck transection. After 1 hour of ischemic circulatory arrest, the perfusion fluid based on autologous blood was continuously perfused through bilateral common carotid artery for 6 hours. The temperature of the perfusion fluid was set at 33 ℃ in the mild hypothermia with perfusion group and 37℃ in the normothermia with perfusion group, respectively. Blood oxygen saturation was recorded at 0, 1, 2, 3, 4, 5 and 6 hours after the beginning of perfusion to evaluate the perfusate oxygen level. The perfusate was collected, and the levels of Na +, K +, Ca 2+ and glucose as well as the pH value of the perfusate were detected in the two groups. At the end of perfusion, the parietal brain tissues of 1 dog from each group were collected to evaluate the water contents of brain tissues. Nissl staining was used to evaluate the morphological integrity of the pyramidal neurons in the frontal cortex and hippocampus. Neuronal nuclei antigen (NeuN) was used to evaluate the structural and morphological integrity of pyramidal neurons. Immunofluorescence glial fibrillary acidic protein (GFAP) and ionic calcium binding adaptor molecule 1 (Iba1) were used to evaluate the integrity and activity of astrocytes and microglia fragments. Results:At 0, 1, 2, 3, 4, 5 and 6 hours of perfusion, there was no significant difference in the blood oxygen saturation or Na + concentrations between the two groups (all P>0.05); the K + concentrations in the mild hypothermia with perfusion group were (4.57±0.12)mmol/L, (4.67±0.14)mmol/L, (4.27±0.12)mmol/L, (4.45±0.10)mmol/L, (6.60±0.15)mmol/L, (7.37±0.18)mmol/L and (9.03±0.16)mmol/L, respectively, which were significantly lower than those in the normothermia with perfusion group [(4.84±0.10)mmol/L, (5.31±0.13)mmol/L, (5.44±0.24)mmol/L, (5.70±0.18)mmol/L, (7.79±0.18)mmol/L, (10.44±0.40)mmol/L, (10.40±0.41)mmol/L] (all P<0.01). At 0, 1, 2 and 3 hours of perfusion, the Ca 2+ concentrations in the mild hypothermia with perfusion group were (0.72±0.15)mmol/L, (1.55±0.16)mmol/L, (1.62±0.15)mmol/L and (1.88±0.15)mmol/L, respectively, being significantly higher than those in the normothermia with perfusion group [(0.41±0.13)mmol/L, (0.99±0.12)mmol/L, (1.29±0.13)mmol/L, (1.57±0.11)mmol/L] (all P<0.01), and no significant differences were found at other time points (all P>0.05). At 0, 1 and 2 hours of perfusion, the glucose concentrations in the mild hypothermia with perfusion group were (5.75±0.19)mmol/L, (5.17±0.15)mmol/L and (4.72±0.15)mmol/L, respectively, being significantly higher than those in the normothermia with perfusion group [(5.30±0.22)mmol/L, (4.89±0.20)mmol/L, (4.30±0.17)mmol/L] (all P<0.01), with no significant differences found at other time points (all P>0.05). At 2, 3, 4, 5 and 6 hours of perfusion, the pH values of the mild hypothermia with perfusion group were 7.32±0.06, 7.25±0.02, 7.23±0.02, 7.24±0.02 and 7.24±0.02, respectively, being significantly higher than those in the normothermia with perfusion group (7.26±0.01, 7.21±0.01, 7.17±0.02, 7.15±0.02, 7.08±0.02) ( P<0.05 or 0.01), with no significant differences at other time points (all P>0.05). The water content of brain tissues in the mild hypothermia with perfusion group was (74.9±0.4)%, which was significantly lower than (79.9±0.9)% in the normothermia with perfusion group ( P<0.01). Nissl staining showed that the pyramidal neurons in prefrontal cortex and dentate gyrus had good integrity in the mild hypothermia with perfusion group. NeuN immunofluorescence staining showed that the morphology and structure of pyramidal neuron cells in the mild hypothermia with perfusion group were better with clearly visible axons than those in the normothermia with perfusion group, whereas the cytosol was full and swollen with scarce axons in the normothermia with perfusion group. GFAP and Iba1 immunofluorescence staining showed that more structurally intact glial cells, more abnormally active cells, thickener axons and better axon integrity in all directions were found in the mild hypothermia with perfusion group than those in the normothermia with perfusion group. Conclusion:Compared with normal temperature antegrade mechanical perfusion, the mild hypothermia antegrade mechanical perfusion can protect canine brain tissue and alleviate ischemic brain injury by maintaining stable energy and oxygen supply, balancing ion homeostasis and perfusion fluid pH value, reducing tissue edema, and maintaining low metabolism of pyramidal neurons, astrocytes and microglia.
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Objective To observe the efficacy and safety of recombinant human growth hormone ( rhGH) replacement therapy in GHD childhood with craniopharyngioma after surgery. Methods This study retrospectively reviewed the records of 18 inpatients with secondary GHD diagnosed by insulin tolerance test ( ITT ) after craniopharyngioma surgery at the Department of Endocrinology, Beijing Tiantan Hospital, from January 2012 to December 2015. The clinical benefits and risks of 18 patients were evaluated systematically, and then were divided into rhGH treatment group(n=9) and control group(n=9). The parameters of height, growth velocity(GV), height standard deviation score (HtSDS), insulin-like growth factors-1 (IGF-Ⅰ), insulin-like growth factor binding protein 3 (IGFBP3) and adverse events rate were recorded after treatment for six months. MRI was followed up every 3 to 6 months to observe the difference of the tumor recurrence and second malignant neoplasm between two groups. Results All 18 patients with craniopharyngioma presented with multiple pituitary-target glands hormone deficiency after surgery. Among these patients, 17 cases (95% ) presented with hypothyroidism or adrenal insufficiency, 7 cases (39% ) with delayed puberty, and 12 cases(67% ) with central diabetes insipidus. Based on pituitary-target gland axis function deficiency, these patients were given appropriate L-thyroxine, prednisone, and desmopressin(DDAVP) replacement therapy, respectively. The median time of 9 patients starting rhGH replacement was 48(36,72)months after surgery. The levels of height, GV, IGF-Ⅰ, HtSDS, and IGFBP3 were significant increased after rhGH treatment for 6 months as compared with pre-treatment and control group (all P 0. 05 ). The tumor recurrence and second malignant neoplasm were not detected by MRI scanning in rhGH treatment group,but there were 3 cases in the control group. Conclusion Multiple pituitary-target glands axis deficiencies were observed in childhood patients with craniopharyngioma after neurosurgery, and the evident deficiency of GH-IGF-Ⅰ axis was observed. rhGH replacement therapy in short-term would significantly improve the parameters of growth and development of patients with GHD after craniopharyngioma neurosurgery. No recurrence tumor in situ and second malignant neoplasms were detected during the period of rhGH replacement therapy.
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Objective To explore the impact of silver ions on the adhesion ability of Bacillus Calmette-Gueri vaccine on silicone tube surface. Methods Common and silver plated silicone tubes were cultured in BCG bacterial liquid for 30 days. A gas-bacilli was taken once every 3 days to measure OD value of microbial. The growth curve of BCG vaccine was made. The corresponding silicone tubes in divided different generations was removed and eluted based on the division cycle of BCG vaccine. The samples were inoculated on the solid medium for observing the colony growth amount, and adhesion curve was drawn. The silicone tube colony line was observed under the electron microscope at day 30. Results OD value and bacterial plaque number were less in silver silicone tube group than in common silicone tube group, with statistical significances (P < 0.05). Electron microscopy also showed that BCG adhesion on silver silicone tube group was relatively less than that on common silicone tube group. Conclusion Silver ions can reduce the adhesion of BCG vaccine on silicone tube surface and inhibit the growth of BCG vaccine.
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Objective To discuss the microanatomy features of jugular foramen region in transpetrosal-presigmoidal minimally invasive approach based on virtual reality image models.Methods CT and MRI scans were performed to fifteen adult cadaver heads,and then,imaging data were inputted into Vitrea virtual reality system to establish three-dimensional anatomy models of jugular foramen region.Different minimally invasive transpetrosal-presigrnoidal approaches exposing superior edge of internal acoustic meatus (route A),anterior edge of jugular foramen (route B),and posterior edge of jugular foramen (route C) were simulated by selecting osseous landmark points.Anatomic exposures in surgical trajectory following alternation of minimally invasive approach were observed,measured and compared.Results Spacial sequence of nerves and vessels in the route simulating transpetrosal-presigmoidal approach for exposure of jugular foramen region was displayed clearly;route A had the highest location and route C was the lowest.Volumes of route and petrosal osseous structure involved in route A was the largest,and then,volumes of route and osseous structure involved in route B was larger than those in route C,with significant differences (P＜0.05).Volumes of involved venous sinus were as follow:route B＞route A＞route C,with significant differences (P＜0.05).Volumes of facial-acoustic nerve complex and anterior inferior cerebellar artery involved in route A were (53.32± 5.54) mm3 and (30.55±3.51) mm3,respectively.Volumes of lower cranial nerves involved in route B were (84.59±9.23) mm3.Conclusion There are different impacts on the osseous structures and venous sinus for the minimally invasive routes of transpetrosal-presigmoidal approach exposing different targets,of which combinations are helpful to utilize respective advantages.
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Objective To observe the effects of fructose,fibroblast growth supplement(FGS) and ethylamine sulfonic acid on the total number,the survival rate and the survival number of Human Retinal Pigment Epithelial(hRPE) Cell.Methods Microencapsulated hRPE cells were plated and cultured in four kinds of mediums,which contained fructose,fibroblast growth supplement(FGS),ethylamine sulfonic acid or no extra ingredient respectively.The total cell number,survival rate and viable cell number of the microencapsulated hRPE cell on day 0th,1st,3rd,7th were calculated.Results After 7days of culture,the lowest cell survival rate of microencapsulated hRPE cells in the four groups was(75.00±3.00)%,but there were no significantly differences(Ps＞0.05) among the groups.The total number of cells in the fibroblast growth factor group([8.00±0.46]×104) and ethylamine sulfonic acid group([7.20±0.36]×104) were significantly higher than the blank group(([6.10±0.56]×104),Ps＜0.05),while no statistical difference was observed in the comparison between the fructose group([6.00±0.46]×104) and blank control(P＞0.05).Conclusion The FGS and ethylamine sulfonic acid can promote the proliferation of the microencapsulated hRPE cells.
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@#ObjectiveTo study a practical individualized computer-aided design project for cranioplasty in Chinese mainland. Methods6 patients with cranial defect was performed superthin skull CT,three-dimension reconstruction. The template for the skull defected was made simulating the shape and contour of cranial defect. after patient,s consent,silicone rubber terylene patch was made as a substrate for the repair of cranial defect. ResultsThis project provides a good result,especial an excellent cosmetic result. No side-effect appears,operation time is shorter than ever.ConclusionAn individual project for computer-assisted cranioplasty has been finished,which can be popularized for its value.
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Objective To investigate the electrophysiological characteristics of the ventralis intermedius nucleus (Vim) in order to find an easy and safe way to confirm the target in Vim-thalamotomy. Method In microelectrode-guided selective Vim-thalamotomy for 23 Parkinson's disease patients, the background activity, amplitude and discharge frequencies of Vim were compared with its surrounding structures. The response of kinesthetic neuron and tremor cell to microstimulation was also compared. Result There were differences in backgroud activity and discharge amplitude for Vim, ventralis lateralis nucleus (VL), ventralis caudalis nucleus (VC), and internal capsule. Based on the response to active or passive movement of contralateral limb tremor cells were divided into two subgroups, which were different in localization. Contralateral tremor showed different response when the two subgroups of tremor cells were mircrostimulated. Conclusion The anterior border of Vim was easily found by microrecording. Only by combining microstimulation with microrecording could the posterior border of Vim and its interior and lateral ordination of target were identified exactly and safely. Kinesthetic neurons and tremor cells which responded to the movement of contralateral limbs should be destroyed.
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Objective To summarize the clinical effect of individualized cranioplasty with different materials. Method The clinical data of 75 patients with skull defect were analyzed retrospectively. Ultrathin cranial CT and three-dimension reconstruction were done to delineate the shape and contour of cranial defect. The templet for repair was fabricated with methyl silicone rubber, bone cement or titanium mest according to the condition of the bone defect with laser rapid prototyping technique. With patient's consent, the individualized templet was implanted to repair the cranial defect. Results Methyl silicone rubber or bone cement were used for 40 of 75 patients to fill the defects, and in the other 35 patients titanium mesh was used to cover the defects with overlapping edges. The patches matched cranial defect perfectly without the necessity of revision. The average operation time was 45 minutes. Perfect or excellent cosmetic results was obtained in 63 of the 75 patients (84%). Postoperative complications were as follows: 5 patients complained of headache, which disappeared in there months. Collection of fluid under the scalp was found in 12 patients, among them the fluid was Absorbed 10 days after operation in 8 cases without any treatment, while in 3 patients it disappeared after suction for 1-4 times. In the remaining one patient, the bone cement templet was removed due to infection, and a titanium mesh was implanted one year later. Loosening and displacement of the templet occurred in 2 patients, and silk sufures were used for fixation in both patients. In these two patients, the templet became stable 3 months later, and in the other titanium was used for repair one year later. The incidence of complications was signifieantly different between inlay and onlay methods of repair (P