Objective To study the changes of proteome expression in brain tissues from rats with severe traumatic brain injury(sTBI). Methods Total protein of brain tissues were obtained at days 3,7 and 14 for two-dimensional gel electrophoresis to screen and identify differential protein spots.Results We screened 17 differential protein spots that were involved in cellular metabolism,stress and inflammatory reaction. Conclusion Some differential proteins involved in sTBI can be found by twodimensional gel electrophoresis.

Objective To locally inject human umbilical cord blood (HUCB) mesenchymal stem cells (MSCs) to rat traumatic brain injury (TBI) model to investigate expression of neural markers and neurological functional improvement. Methods HUCB-MSCs were labeled by bis-benzimide for over 24 hours and stereotactically transplanted into the brain of the rats. All rats were divided into four groups, ie, sham injury group, TBI group, control (TBI + PBS) group and treatment (TBI + MSCs) group, Im-munohistochemical methods and immanofluorescence staining were used to observe the survival, migration and differentiation of the transplanted cells. The neurological functional improvement was evaluated by u-sing the neurological severity score (NSS). Results There existed a large number of MSCs survived in local region of the brain that received transplants, when some MSCs differentiated into neurons or astro-cytes and expressed the neurocyte markers including NSE and GFAP around the grafted site. Treatment group had significantly improved scores compared with sham injury group, TBI group and control group. Conclusions HUCB-MSCs transplantation can potentially improve neurological functional after TBI and may be a good alternative to bone marrow cells for stem cell transplantation or cell therapy.

The experiment was conducted by directed evolution strategy (error-prone PCR) to improve the activity of aflatoxin detoxifzyme with the high-throughput horse radish peroxidas and recessive brilliant green (HRP-RBG) screening system. We built up a mutant library to the order of 10(4). Two rounds of EP-PCR and HRP-RBG screening were used to obtain three optimum mutant strains A1773, A1476 and A2863. We found that mutant A1773 had upper temperature tolerance of 70 degrees C and that its enzyme activity was 6.5 times higher than that of the parent strain. Mutant strains A1476 worked well at pH 4.0 and its enzyme activity was 21 times higher than that of the parent strain. Mutant A2863 worked well at pH 4.0 and pH 7.5, and its enzyme activity was 12.6 times higher than that of the parent strain. With DNA sequencing we found that mutant A1773 revealed two amino acid substitutions, Glu127Lys and Gln613Arg. Mutant A1476 revealed four amino acid substitutions: Ser46Pro, Lys221Gln, Ile307Leu and Asn471lle. Mutant A2863 revealed four amino acid substitutions: Gly73Ser, Ile307Leu, Va1596Ala and Gln613Arg. The results provided a useful illustration for the deep understanding of the relationship between the function and structure of aflatoxin detoxifzyme.
Aflatoxin B1 ; antagonists & inhibitors ; chemistry ; Amino Acid Substitution ; Directed Molecular Evolution ; Enzyme Activation ; Enzyme Stability ; Multienzyme Complexes ; genetics ; metabolism ; Mutant Proteins ; genetics ; metabolism ; Point Mutation ; Polymerase Chain Reaction ; methods ; Protein Engineering

OBJECTIVETo explore the role and function of stromal cell-derived factor-1 (SDF-1) in stem cells migrating into injured brain area.

METHODSRat-derived nerve stem cells (NSCs) were isolated and cultured routinely. Transwell system was used to observe the migration ability of NSCs into injured nerve cells. Immunocytochemistry was used to explore the expression of chemotactic factor receptor-4 (CXCR-4) in NSCs. In vivo, we applied immunofluorescence technique to observe the migration of NSCs into injured brain area. Immunofluorescence technique and Western blotting were used to test expression level of SDF-1. After AMD3100 (a special chemical blocker) blocking CXCR-4, the migration ability of NSCs was tested in vivo and in vitro, respectively.

RESULTSNSCs displayed specific tropism for injured nerve cells or traumatic brain area in vivo and in vitro. The expression level of SDF-1 in traumatic brain area increased remarkably and the expression level of CXCR-4 in the NSCs increased simultaneously. After AMD3100 blocking the expression of CXCR-4, the migration ability of NSCs decreased significantly both in vivo and in vitro.

CONCLUSIONSSDF-1 may play a key role in stem cells migrating into injured brain area through specially combining with CXCR-4.

Animals ; Brain Injuries ; pathology ; Cell Movement ; Cells, Cultured ; Chemokine CXCL12 ; analysis ; physiology ; Neurons ; cytology ; Rats ; Receptors, CXCR4 ; analysis ; physiology ; Stem Cells ; physiology ; Tropism

Objective To establish the immortalized umbilical cord mesenchymal stem cells(UCMSCs) mediated by human telomerase reverse transcriptase (hTERT) so as to offer enough UCMSCs for in vitro or clinical researches. Methods Human UCMSCs were isolated and cultured to passage 4,and then identified by flow cytometry analysis. The pLXSN-hTERT plasmid was transfected into UCMSCs by liposome to establish the immortalized UCMSCs. Expressions of hTERT mRNA and protein were respectively tested by RT-PCR and immunocytochemistry. The cell cycle kinetics of the passage of hTERT-UCMSCs and UCMSCs were detected with flow cytometry. Results Flow cytometry analysis showed that UCMSCs expressed CD29, CD44 and CD 105 strongly, but CD31, CD34 and CD45 slightly; these were right the features of human UCMSCs. RT-PCR showed that hTERT mRNA was expressed low in UCMSCs, and high in pLXSN-hTERT transfected UCMSCs.Immunocytochemistry revealed that fluorescence intensity of cell nuclei was increased significantly after transfection and that hTERT was expressed all in cell nuclei. Flow cytometry analysis suggested that the number of hTERT-UCMSCs at phase S was increased significantly, and the cells would be able to passage stably (more than 35 passages). Conclusions The immortalized UCMSCs can be established by hTERT transfection, and the number of immortalized UCMSCs can meet the need of in vitro or clinical researches.

Objective To evaluate the effect of downregulation of microRNA (miR)-373 expression on cell cycle and apoptosis of a cutaneous squamous cell carcinoma (CSCC) cell line A431.Methods A431 cells at exponential growth phase were classified into 3 groups:miR-373 inhibitor group and negative control group transfected with miR-373 inhibitor and negative control miRNA respectively,and untreated group receiving no treatment.At 48 hours after the transfection,real-time PCR was performed to determine the expression of miR-373 in the above 3 groups,cell counting kit-8 (CCK-8) assay to evaluate the effect of downregulated expression of miR-373 on the proliferation of A431 cells,flow cytometry to investigate the distribution of cell cycle and changes in apoptosis of A431 cells in different treatment groups,and colorimetric analysis to detect the changes in caspase-3 activity in different treatment groups.Statistical analysis was carried out with SPSS 17.0 software by using two-sample t test for the comparison between two groups,one-way analysis of variance (ANOVA) for the comparison among 3 groups,and least significant difference (LSD)-t test for multiple comparisons.Results The expression of miR-373 was significantly lower in the miR-373 inhibitor group (0.120 ± 0.036) than in the untreated group (1.002 ± 0.022) and negative control group (1.037 ± 0.028,LSD-t =36.21,34.83,respectively,both P ＜ 0.001).At 48,72 and 96 hours,the miR-373 inhibitor group showed significantly decreased proliferative activity of A375 cells compared with the untreated group and negative control group (F =10.805,13.720 and 30.907 respectively,P =0.038,0.010 and 0.001 respectively).The proportion of A375 cells in G0/G1 phase was significantly higher in the miR-373 inhibitor group (64.69％ ± 1.18％) than in the untreated group (52.74％ ± 0.66％,t =15.51,P ＜ 0.001) and negative control group (53.80％ ± 0.80％,t =13.24,P ＜ 0.001).The proportion of total apoptotic cells and activity of caspase-3 in the miR-373 inhibitor group were 22.69％ ± 1.24％ and 1.238 ± 0.057 respectively,which were significantly higher than those in the untreated group (9.62％ ± 1.14％,0.413 ± 0.028 respectively,both P ＜ 0.001)and negative control group (9.66％ ± 0.97％,0.437 ± 0.036 respectively,both P ＜ 0.001).Conclusion MiR-373 may play an important role in the regulation of cell cycle and induction of apoptosis of the CSCC cell line A431.

OBJECTIVE: To investigate forensic diagnosis application of three-dimentional reconstruction with spiral computed tomography in fracture of anatomical complicated bones. METHODS: Selected eleven patients of bone fracture who were examined with SCT 3D and conventional X-ray examination. The location, number and characteristics were observed and analyzed. RESULTS: In all of eleven patients with bone fractures, X-ray examination could detect thirty-four rib fracture, one scapula fracture, two nasal fracture, one metacarpal bone incomplete fracture and one left tibia-fibula fracture, one pubis fracture. While there were forty-seven rib fracture, one scapula smash fracture, one nasal fracture with obvious displacement and eliminate one misplace, one left tibia-fibula obsolete fracture and one sacroiliac joint dislocation, one No 5 lumbar vertebrae pedicle of vertebrae arch fracture. Combining 3D reconstruction images, coronary and sagittal reconstruction images could show clearly the fracture line, location of fracture, number of fracture, displacement and recovery. CONCLUSION 3D reconstruction technique of SCT is a very useful examination method in the objective forensic diagnosis of anatomical complicated bones fracture, it excels the routine X-ray examination.
Adult ; Aged ; Female ; Forensic Medicine ; Fractures, Bone/pathology* ; Humans ; Imaging, Three-Dimensional ; Male ; Middle Aged ; Nasal Bone/injuries* ; Rib Fractures/diagnostic imaging* ; Scapula/injuries* ; Tomography, Spiral Computed/methods* ; Young Adult

OBJECTIVETo promote stem cells differentiation into neurons and enhance neuromotor function after brain injury through brain-derived neurotrophic factor (BDNF) induction.

METHODSRecombinant adenovirus vector was applied to the transfection of BDNF into human-derived umbilical cord mesenchymal stem cells (UCMSCs). Enzyme linked immunosorbent assay (ELISA) was used to determine the secretion phase of BDNF. The brain injury model of athymic mice induced by hydraulic pressure percussion was established for transplantation of stem cells into the edge of injury site. Nerve function scores were obtained, and the expression level of transfected and non-transfected BDNF, proportion of neuron specific enolase (NSE) and glial fibrillary acidic protein (GFAP), and the number of apoptosis cells were compared respectively.

RESULTSThe BDNF expression achieved its stabilization at a high level 72 hours after gene transfection. The mouse obtained a better score of nerve function, and the proportion of the NSE-positive cells increased significantly (P<0.05), but GFAP-positive cells decreased in BDNF-UCMSCs group compared with the other two groups (P<0.05). At the site of high expression of BDNF, the number of apoptosis cells decreased markedly.

CONCLUSIONBDNF gene can promote the differentiation of the stem cells into neurons rather than glial cells, and enhance neuromotor function after brain injury.

Adenoviridae ; genetics ; Animals ; Apoptosis ; Brain Injuries ; physiopathology ; therapy ; Brain-Derived Neurotrophic Factor ; analysis ; genetics ; Cell Differentiation ; Glial Fibrillary Acidic Protein ; Humans ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; cytology ; Mice ; Nerve Tissue Proteins ; analysis ; Neurons ; cytology ; Phosphopyruvate Hydratase ; analysis ; Transfection

Objective:To explore the effect of the care and nursing practice plan based on the timing theory in primary caregivers of infants with infantile spasms.Methods:From July 2019 to June 2020, 80 infants with infantile spasm and 80 primary caregivers in the Department of Neurology of Hunan Children's Hospital were selected as the research object. All infants with infantile spasms were numbered according to the time of admission. The odd-numbered infants were admitted to the First Department of Neurology as the observation group, and the even-numbered children were admitted to the Second Department of Neurology as the control group, with 40 infants and 40 primary caregivers in each group. The control group carried out conventional treatment and nursing, and the observation group implemented a care and nursing practice plan based on the timing theory on the basis of the control group. Before and after the intervention, the Caring Ability Inventory (CAI) , Family Caregiver Task Inventory (FCTI) , Caregiver Burden Inventory (CBI) , and Gesell Developmental Schedules (GDS) were used to evaluate the caring ability, caregiving ability, care load of the primary caregivers of the two groups of infants, and the cognition function of the two groups of infants.Results:After the intervention, the total CAI scores of the primary caregivers in the observation group were higher than those before intervention and those in the control group after the intervention, and the total scores of FCTI and CBI were lower than those before intervention and those in the control group after the intervention, and the differences were statistically significant ( P<0.05) . After the intervention, the developmental quotient (DQ) values of the five energy areas of GDS in the observation group were higher than those before the intervention, and the DQ values of the three energy areas of gross motor, fine motor, and language were higher than those of the control group after the intervention, and the differences were statistically significant ( P<0.05) . Conclusions:The care and nursing practice plan based on the timing theory can improve the caring ability and caregiving ability of the main caregivers of children with infantile spasms, reduce the care load, and improve the cognitive function of the children.

OBJECTIVETo investigate the association between the tissue inhibitor of metalloproteinase-2 (TIMP-2) gene polymorphisms with the predisposition and disease severity of thoracic adolescent idiopathic scoliosis (AIS).

METHODSThree hundred and fifty-four female thoracic AIS patients treated from January 2007 to March 2009 and 210 healthy female who underwent health examination during March 2005 to June 2006 as normal controls were recruited in this study. One SNP-418G/C (rs8179090) in the promoter region were selected for TIMP-2 gene. PCR- RFLP was used for genotyping.

RESULTSNo significant differences of genotype and allele frequency distribution were found between AIS patients and normal controls (P > 0.05). In AIS patients, the frequency of C allele of the patients with the body mass index (BMI) < 17 kg/m(2) was significantly higher than those with the BMI > or = 17 kg/m(2) (P < 0.05), and the frequency of C allele of cases with the major Cobb angle > or = 40 degrees was significantly higher than that with Cobb angle < 40 degrees (P < 0.05). Among the patients who reached skeletal maturity without any interference of natural history, significantly higher average maximum Cobb angle was found in patients with GC and CC genotype compare with those with GG genotype.

CONCLUSIONSThe SNP-418G/C (rs8179090) in the promoter region of TIMP-2 gene may be associated with abnormal growth pattern and curve progression of thoracic AIS. TIMP-2 gene is a disease-modifier gene of thoracic AIS.

Adolescent ; Case-Control Studies ; Female ; Genetic Predisposition to Disease ; Genotype ; Humans ; Polymorphism, Genetic ; Polymorphism, Single Nucleotide ; Scoliosis ; genetics ; Tissue Inhibitor of Metalloproteinase-2 ; genetics