Abstract In order to repair the bone defect afteroperation of benign lesion of extremity, the fetal demineralized bone was applied in 10 cases. These cases were followed up for 6 months to 8 years. The results showed that the grafted bone was integrated with the host bone in 6 months. Noadverse effect was found. The demineralized bone did not induce rejection. The advantages of using fetal demineralized bone were as follows: easily obtainable,its preparation and method of storage simple, and low finacial cast.
OBJECTIVE: To explore the effects of different methods of fetal spinal cord(FSC) tissue transplanted on reversing the axotomy-induced neurons atrophy of adult rats injured spinal cord. METHODS: One hundred and twenty adult rats received lumbar spinal cord hemisection. Experimental rats were divided into five groups, the control group(Group A); spinal cord hemisection only(Group B); spinal cord hemisection plus FSC transplant (Group C); spinal cord hemisection plus FSC transplant plus pedicled paraspinal muscle(Group D); spinal cord hemisection plus FSC transplant plus pedicled omentum (Group E). Combined behavioral scores(CBS), somatosensory evoked potentials (SEP), motor evoked potentials(MEP) were examined to evaluate the recovery of neurological function after operation. Rats were sacrificed after 1, 4 and 12 weeks. Nissl stained section was used for neurons quantitative image analysis. The positive cells were quantitative analysis by computer image analysis system. RESULTS: The different methods of FSC tissue transplantation could prevent the neurons atrophy secondary to axon injury of spinal cord in adult rats. The size of neurons were observed in five groups, they were group E gt; group D gt; group C gt; group B gt; group A (P lt; 0.05). Those increases in size of neurons were paralleled with a significant improvement in neurological function recovery. CONCLUSION: It indicates that the different methods of FSC tissue transplantation can maintain the neurons morphology and improve the neurological function of rats.
OBJECTIVE: To investigate the repairing effect of transplantation of allogeneic fetal bone in combination with a covering cryopreserved periosteal allograft to bone defect. METHODS: Twenty Long-eared white male rabbits were chosen as experimental model of bilateral 12 mm combined bony and periosteal radial defect. Cryopreserved allograft periosteum with allogeneic fetal bone were implanted in the left defect as experimental side and fetal bone was simply transplanted in the right defect as control side. Bone repair process in the two groups were compared by macroscopy, microscopy, roentgenograms and the contents of calcium and phosphate in the defect area at 2, 4, 8 and 12 weeks after transplantation. RESULTS: There was significant statistic difference in the contents of calcium and phosphate between the experimental and control sides at 4, 8 and 12 weeks after transplantation (P lt; 0.05). With time passing by, the contents of calcium and phosphate have the increasing trends. In the experimental group, lamella bone was seen and medullary canal recanalized at 8 weeks postoperatively. The histological section showed the bone lacuna and lamella bone were formed. CONCLUSION: It suggests that allogeneic fetal bone in combination with a covering cryopreserved periosteal allograft can promote bone repair, and allogeneic fetal bone is excellent bone substitute.
Objective To investigate the possibility of culturing human oral keratinocyte using autologous serum in order to provide theoretical and technical foundation for clinical application of tissue engineering oral mucosa epithelium.Methods The human oral keratinocytes were cultured by the medium containing different concentrations of autologous serum(10%,20%,30%)and fetalbovine serum (10%), respectively. The growth conditions for the cell and the mucosa epithelium in the groups were observed, the cell growth curves were drawn, and the population doubling time (PDT) was counted. Results The results showed that the human oral keratinocyte could proliferate well in the medium containing autologous serum or fetal bovine serum. The differences in the 24hour clone rate and PDT were not significant. Both the area and the thickness of the cultured oral epithelium increased with the increase of the autologous serum concentration, and the difference between autologous serum and fetal bovine serum was significant, especially with the medium containing 20% autologous serum( P＜0.05) . The human nature of the cultured epithelium was demonstrated by the immunofluorescent mouse anti-HLA antigen. Conclusion The autologous serum can replace the fetal bovine serum to culture the oral keratinocyte well, and the cultured oral mucosa epithelium can be better differentiated in the autologous serum than in the fetal bovine serum.
Fetal electrocardiogram (ECG) signals provide important clinical information for early diagnosis and intervention of fetal abnormalities. In this paper, we propose a new method for fetal ECG signal extraction and analysis. Firstly, an improved fast independent component analysis method and singular value decomposition algorithm are combined to extract high-quality fetal ECG signals and solve the waveform missing problem. Secondly, a novel convolutional neural network model is applied to identify the QRS complex waves of fetal ECG signals and effectively solve the waveform overlap problem. Finally, high quality extraction of fetal ECG signals and intelligent recognition of fetal QRS complex waves are achieved. The method proposed in this paper was validated with the data from the PhysioNet computing in cardiology challenge 2013 database of the Complex Physiological Signals Research Resource Network. The results show that the average sensitivity and positive prediction values of the extraction algorithm are 98.21% and 99.52%, respectively, and the average sensitivity and positive prediction values of the QRS complex waves recognition algorithm are 94.14% and 95.80%, respectively, which are better than those of other research results. In conclusion, the algorithm and model proposed in this paper have some practical significance and may provide a theoretical basis for clinical medical decision making in the future.
Abstract: Objective To investigate the influence of cryopreservation on cellular viability of latepregnancy fetal valved allografts in human. Methods The fetal valved allografts with gestational ages ranged from 24 to 40 weeks were sterilely procured within 6 hours after brain death. Each sample was bisected into control group and experiment group. The cellular viability of control group was directly tested and that of experiment group was examined after being storaged in liquid nitrogen for a week through a programmed frozen procedure. The light microscopy, tissue culture and Methylthiazol tetrazolium assay (MTT assay) were used to determine the cellular viability. Results Twelve latepregnancy fetal valved aortic allografts were procured. Light microscopy showed the integrity of the basic structure of the thawed aorta, the normal structure of the collagen and elastic fibers, with part of vascular endothelium lost. There were lots of cells deriving from both groups,but the cellular growing rate of the experiment group was relatively slower. At 490 nm, MTT assay valve of control group was 0.442±0.046, and that of experiment group was 0.424±0.041. The difference between two groups failed to statistically significance(t=1.617, P=0.328). Conclusion There were viable cells in latepregnancy fetal valved allografts after cryopreservation.
OBJECTIVE: To explore the potential possibility of synaptic connection and 3D adhesion between fetal spinal cord cell suspension (FSCS) and host, and to observe the synapses developing process of FSCS transplantation. METHODS: Spinal cord injury model produced in 42 Wistar rats on T7 by use of modified Allen’s impact method (10 g x 5 cm); 3 days after injury, 20 microliters FSCS with a density of 1 x 10(5)/microliter prepared from E14 rat were injected into the epicenter of the traumatized cavity. Animals were sacrificed after 2, 4, 6, 8, 10 and 12 weeks of transplantation, the graft survival, its differentiation and integration with the host were observed by light and electronmicroscopic study as well as immunohistochemical assay (NF, GFAP, CGRP, 5-HT). RESULTS: In the transplantation area, the neuroblasts stretched out the terminal endings 4 weeks after implantation, followed by the presenting of the pre- and postsynaptic membrane. After 8 weeks, the dense or developed projections were observed in the pre- and postsynaptic membrane; the synaptic cleft filled with the high electron dense substance. All the spherical clear vesicles, granular vesicles, elliptical vesicles and flattened-f type vesicles were seen under the electronmicroscope. After 10 weeks, the axosomatic, dendrosomatic, dendro-dendritic, axo-axonic, dendro-axonic synapses coexisted. Light microscopy showed that the graft cell grew gradually. Immunohistochemical assay showed that NF, 5-HT, CGRP and GFAP positive fibers were in the graft. Synapses, gliafibers and blood brain barrier integrated each other. CONCLUSION: (1) The transplanted FSCS can develop mature synapses with miscellaneous synaptic vesicles in the acute injured spinal cord, host injury cavity wall may induce the FSCS into 3D adhesion. (2) Co-existence of different type of synapse and the immunohistochemistry findings indicate the possibility of synaptic connection between FSCS and host.
Objective To explore the change tendency of hypoxia-inducible factor-1α (HIF-1α) and extracellular signal-regulated kinase 1/2 (ERK1/2) in fetal rat cerebral cortex neurons cultured in vitro after hypoxia-ischemia reperfusion andto investigate their mutual relationship. Methods Cortical neurons obtained from cerebral cortex of 15 pregnant SD rats at16-18 days of gestation underwent primary culture. The primary neurons 5 days after culture were adopted to establ ish model of oxygen and glucose deprivation (OGD). The experiment was divided into 4 groups: the experimental group 1, culture medium was changed to neuron complete medium containing glucose after the preparation of OGD model to form reperfusion, and the neurons were observed 0, 2, 4, 8, 12 and 24 hours after reperfusion; the control group 1, the neurons were treated with normal medium; the experimental group 2, the neurons were pretreated with U0126 followed by the preparation of OGD model, and the neurons were observed 4 and 8 hours after reperfusion; the control group 2, the neurons were pretreated with DMSO, and other treatments were the same as the experimental group 2. Expressions of HIF-1α, VEGF protein, ERK1/2 and p-ERK1/2 were detected by Western blot. Expression and distribution of p-ERK1/2 and HIF-1α protein were detected by SABC immunocytochemistry method. Results Compl icated synaptic connections between cortical neurons processes were observed 5 days after culture. The expression of HIF-1α and VEGF were increased gradually, peaked at 8 hours, and decreased gradually after 12 hours in the experimental group 1, and there were significant differences between the experimental group 1 and the control group 1 (P lt; 0.05). There was no significant difference between the experimental group 1 and the control group 1 in terms of ERK1/2 protein expression (P gt; 0.05). The p-ERK1/2 protein expression in the experimental group 1 started to increase at 2 hours peaked at 4 hours, and started to decrease at 8 hours, showing significant differences compared with the control group 1 (P lt; 0.01). In the experimental group 2, the p-ERK1/2 protein decreased, and HIF-1αand VEGF protein expression subsequentlydecreased, showing significant differences compared with the control group 2 (P lt; 0.05). There was no significant difference between the experimental group 2 and the control group 2 in terms of ERK1/2 protein expression at each time point (P gt; 0.05). Immunocytochemistry staining showed that p-ERK1/2 and HIF-1α expression decreased, and the yellow-brown staining of the neurons was reduced. Conclusion Expressions of HIF-1α and its target-gene VEGF protein in the cortex neurons after OGD reperfusion are time-dependent. Their expressions decrease when ERK1/2 signal ing pathway is inhibited, indicating the pathway plays an important role in the regulation of HIF-1α and VEGF induced by OGD of cortical neurons
Objective?To study the mid-facial development characteristics of the goats with cleft palate after in-utero surgical repair at different stages.?Methods?Twenty-four Boer hybrid female goats were selected, aged from 8 to 12 months and weighing from 35 to 55 kg. The mating day was designated for 0 day. At 30 days, pregnant was confirmed by B-ultrasound test, and the goats were divided into 5 groups (experimental groups 1, 2, 3, 4, and normal control group). Twenty pregnant goats of 4 experimental groups (n=5) were injected DL-anabasine (15 mg/day) from 31 to 42 days to establish cleft palate model of fetal lamb, 4 pregnant goats of normal control group used as controls without injection. At pregnant 65, 90, and 120 days, cleft palate was repaired in the uterus in experimental groups 1, 2, and 3, while cleft palate was not repaired in experimental group 4. After 1 month of birth, the maxillary bone width (posterior premolar morphological measurement, PPMM) and the maxillary bone length (anterior premolar morphological measurement, APMM) were measured with CT scanning. The dry skull of goats were harvested for gross observation.?Results?There was no significant difference in PPMM and APMM between experimental group 1 and the normal control group (P gt; 0.05), but there were significant differences between experimental groups 1 and 4 (P lt; 0.05) at 1 month after birth. Significant differences were oberved in PPMM and APMM between experimental group 2 and normal control group, experimental group 4 (P lt; 0.05). There were significant differences in PPMM between experimental group 3 and normal control group, experimental group 4 (P lt; 0.05), in APMM between experimental group 3 and normal control group (P lt; 0.05). Five goats with cleft palate in experimental group 4 died at 1-2 months after birth.?Conclusion?At pregnant 65 days, in-utero surgical repair of cleft palate has less influences on mid-facial development. The earlier repair is performed, the higher risk of miscarriage was.
Objective To investigate effects of neural retina on development of the structure of outer blood retinal barrier in embryogenesis. Methods The retinal neural epithelium (RNE) and pigment epithelium (RPE) layers of 150, 120 and 90 embryonic chicken eyes incubated for 7,10, and 14 days were peeled off. RNE was used to prepare the culture medium with different conditions (7drcSF3, 10drcSF3, 14drcSF3). RPE cells of 7- and 14-incubated chicken embryos were cultured on laminin-coated transwell filter. The SF3, 7drcSF3, 10drcSF3 , 14drcSF3 medium were used respectively in the apical chamber and SF2 was used in basolateral chamber. After the formation of monolayer, the transepithelial electrical resistance of the RPE was detected. After the fixation of RPE cells, the condition of the tight junction among the cells was observed by immunohis tochemistry and transmission electron microscopy. Results For the RPE cells of 7-and 14-day incubated embryonic eyes, the difference of TER in various medium of SF3/SF2, 7drcSF3/SF2, 10drcSF3/SF2, 14drcSF3/SF2 was statistically significant (P＜0.01). The polarity of RPE cells was induced and the netlike tight junctional strands was urged in the retina-conditioned medium. Conclusion The neural retina may actively promote the formation of the structure of outer blood retinal barrier. (Chin J Ocul Fundus Dis，2004，20：237-240)