Objective To study the feasibility of stem cell transplantation under mild hypothermia so as to provide a prerequisite for stem cell transplantation in patients with traumatic brain injuries (TBI) during mild hypothermia treatment. Methods After transfecting plasmid containing temperature-sensitive simian virus 40 large T-antigen (tsSV40LT) into temperature-sensitive umbilical cord mes-enchymal stem cells (tsUCMSCs) , the changes of cell morphology, nuclear proliferation index (PIx) and telomerase activity were detested when the tsUCMSCs were cultured at 33℃ and 37℃. After the mouse model with tTBI treated with mild hypothermia was established, tsUCMSCs were transplanted into semi-injury area to detect survival rate, proliferation and apoptosis indices and perform neurological deficit scoring. Results When cultured at 33℃, the tsUCMSCs displayed long spindle-shaped and highly refractive, with higher proliferation index and telomerase activity than those cultured at 37℃. Compared with control group (non-temperature-sensitive UCMSCs transplantation), tsUCMSCs in semi-injury area showed much higher cell survival and proliferating cell nuclear antigen expression ( P < 0.05 ) , with fewer apoptotic cells and better neurological function (P < 0.05). Conclusion The establishment of temperature-sensitive stem cell line enables stem cell transplantation during treatment of TBI with mild hypothermia, as provides us a new direction for treatment of TBI.

Human enhancement technology is an emerging field that aims to enhance human physical and mental power of humans with the latest developments of the life science , information science , electronic science , cognitive neuroscience and other fields .This paper provides an overview of status quo of human enhancement technology and its military applica -tions, and analyzes the factors that may adversely affect its applications .

2013 saw sustained and rapid development in the military medicine-related life sciences .New research fields, new technologies and devices continue to emerge , such as Brain Science Projects , 3D bio-printing, cognitive neuro-science, brain-computer interface and mind control , genome editing technology , neuroimaging techniques , protein research technology , Which will promote the all-round development of basic and applied research for military medicine .

ObjectiveTo analyze the distribution and antibiotic resistance of bacteria isolated from cerebrospinal fluid. Methods569 cerebrospinal fluild specimens were analyzed. ResultsPathogens were isolated from 93 specimens (16.3%), in which 52 were Gram positive bacteria and 38 were negative, 3 were fungi. All the Gram positive cocci were sensitive to vancomycin and linezolid, while the Gram negative bacteria were resistant to routine antibiotics such as cefotaxime and ceftriaxone, but sensitive to carbapenems. ConclusionThe prevailing pathogens in intracranial infection are Gram positive cocci, especially Staphylococcus epidermidis and Staphylococcus aureu.

Objective To judge injury severity of severe traumatic brain injury (sTBI) by using surface enhanced laser desorption-ionization (SELDI) protein chip technique. Methods Serum sam-ples from sTBI patients were used to detect expression of differential proteins by protein chip CM10 and SELDI to analyze the correlation between expression peak intensity and GCS. Results We obtained 101 protein peaks, with statistical difference upon expression of 27 protein peaks, when negative correla-tion was found between two peaks ( m/z 4 972 and m/z 5 322 ) and GCS score and positive correlation be-tween six peaks (m/z 3 941, m/z 4 295, m/z 8 714, m/z 8 792, m/z 14 020 and m/z 28 148) and GCS score. Conclusion SELDI protein chip technique may become a new and objective detection method in judging injury severity of sTBI.

We studied the immunogenicity of pseudorabies virus gC DNA vaccination by fusing the murine complement C3d receptor binding domain. First, pseudorabies virus gC gene was linked to four copies of C3d receptor binding domain (M284), and then cloned into the vector pcDNA3.1 to construct the recombinant plasmid sgC-M284. Through the experiment of immunized BALB/c mice, we found that the enzyme linked immunosorbent assay (ELISA) antibody titer for sgC-M284 was 17-fold higher than that for sgC alone, and protective rate of mice was augmented from 25% to 88% after lethal dose PrV (316 LD50) challenge. In addition, the IL-4 levels for sgC-M284 immunization approached that for the pseudorabies virus inactivated vaccine. In conclusion, we demonstrated murine C3d receptor binding domain fusion significantly increased Th2-biased immune response by inducing IL-4 production.
Adjuvants, Immunologic ; physiology ; Animals ; Antibody Formation ; immunology ; Binding Sites ; Cloning, Molecular ; Complement C3d ; genetics ; immunology ; Herpesvirus 1, Suid ; genetics ; immunology ; Interleukin-4 ; immunology ; Mice ; Mice, Inbred BALB C ; Pseudorabies Vaccines ; immunology ; Receptors, Complement 3d ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; immunology ; Swine ; Vaccines, DNA ; immunology ; Viral Envelope Proteins ; pharmacology ; Viral Fusion Proteins ; immunology

We wished to undertake molecular genetic typing and evaluate recombinants of the hepatitis-B virus (HBV) in Tibet (China). Multistage random sampling was used to collect HBsAg-positive samples. Nested polymerase chain reactions were used to amplify the whole sequence of the HBV. DNAstar, MEGA6 and SimPlot were used to assemble sequences, create phylogenetic trees, and undertake recombination analyses. Twelve whole sequences of the HBV of a Tibetan population were collected using these methods. Results showed that all 12 strains were C/D recombinants. Nine of the recombinations were at nt750, and the other three at nt1526. Therefore, the 12 strains could be divided into two types of recombinants: C/Da and C/Db. Analyses of the sequence of the whole genome revealed that the 12 strains belonged to genotype C, and that the nucleotide distance was > 4% between the 12 strains and sub-genotypes C1 to C15 in Genbank. The most likely sub-genotype was C1. Individuals with C/Da were from central and northern Tibet (e.g., Lasa, Linzhi, Ali) and those with C/Db recombinants were from Shannan in southern Tibet. These data suggest that the two types of recombinants had a good distribution in Tibet. Also, they can provide important information for studies on HBV recombination, gene features, virus evolution, as well as the control and prevention of HBV infection in Tibet.
Adult ; Female ; Genotype ; Hepatitis B ; virology ; Hepatitis B virus ; classification ; genetics ; isolation & purification ; Humans ; Male ; Phylogeny ; Recombination, Genetic ; Sequence Analysis, DNA ; Tibet ; Young Adult

We constructed a recombinant plasmid encoding VP1 gene of O type foot-and-mouth disease virus fused to a molecular adjuvant, goat complement C3d gene. The goat C3d gene was cloned and three copies were tandem-linked with the linker (G4S)2 sequence. VP1 gene of O type foot-and-mouth disease virus was linked to three tandem repeats of C3d through the linker sequence and cloned into pUC19 to obtain the recombinant plasmid pUC19-VP1-C3d3. The VP1-C3d3 fusion gene was then subcloned into the eukaryotic vector pcDNA3.1(+) that had been modified to contain the tissue plasminogen activator (tPA) leader sequence to obtain pcDNA3.1-tPA-VP1-C3d3. HeLa cells were transfected with pcDNA3.1-tPA-VP1-C3d3 by Lipofectamine 2000. Indirect immunofluorescent assay and Western blot assay showed that VP1-C3d3 fusion gene was successfully expressed in HeLa cells. The fusion protein with the expected size 133 kD could be secreted outside the cells. This study laid a good foundation to further research on the novel vaccine against foot-and-mouth disease virus by using goat C3d as a molecular adjuvant to enhance the immunogenicity of VP1.
Animals ; Capsid Proteins ; biosynthesis ; genetics ; Cloning, Molecular ; Complement C3d ; biosynthesis ; genetics ; immunology ; Female ; Foot-and-Mouth Disease Virus ; genetics ; Goats ; HeLa Cells ; Humans ; Immunologic Factors ; biosynthesis ; genetics ; immunology ; Plasmids ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology ; Transfection

Objective: To obtain the optimum of lentiviral library packaging based on CRISPR/cas9 (Clustered regularly interspaced short palindromic repeat sequences/CRISPR-associated protein 9). Methods: Reverse transcription polymerase chain reaction (RT-PCR), immunofluorescence antibody (IFA) and enzyme linked immunosorbent assay (ELISA) were used to detect the lentivirus titers in condition of different ratio of packaging plasmids, different addition of lipofectamine 3000 reagent and different time points post-transfection. Then, high-throughput sequencing was performed to evaluate the representation and distribution of single guide (sg)RNAs in the library. Results: The lentivirus titer was the highest when the molar ratio of psPAX2∶pMD2.0G∶Lentivirus library was 2∶1∶1, and the optimum addition of Lipofectamine 3000 reagent was 10 μl, while the result of ELISA were correspondent to that of RT-PCR. The IFA result showed that the lentivirus titer was the highest at 60 h post-transfecion. The coverage of sgRNAs in the lentivirus library packaged with the optimum we obtained was 99.3%, and the read counts of sgRNAs was observed in a normal distribution. Conclusions The optimal lentivirus library packaging was obtained, and this can provide basis for CRISPR/cas9-based screening.

Objective:To investigate the optimal monochromatic level for evaluation of in-stent lumen after transjugular intrahepatic portosystemic shunt (TIPS) by dual-layer detector CT.Methods:Twenty-nine patients after TIPS were retrospectively enrolled who underwent abdomen enhanced examinations with portal venous phases by a dual-layer detector CT between December 2019 and July 2021. The mixed iterative image (conventional group) and monochromatic images (40 keV group, 50 keV group, 60 keV group and 70 keV group) were obtained by reconstruction. Circular regions of interest were placed in the in-stent of the cross-sectional reconstructed image and in the vertical spinal muscle on the same plane to obtain the corresponding average CT value and noise. The contrast to noise ratio (CNR) and signal to noise ratio (SNR) were calculated. Then 4-point scale was performed to evaluate image quality subjectively by 2 physicians blindly and separately. One-way ANOVA or Kruskal Wallis H rank-sum test was used for the overall analysis between groups, and LSD test or Dunn′s Bonforoni test was used for pairwise comparison within groups. Results:There was no significant difference in noise values among the 5 groups ( P>0.05). The difference of CNR and SNR between the 5 groups was statistically significant ( F=72.28, 56.45, P<0.001). The CNR and SNR in the 40 keV group were the highest, which were 50.4±15.7 and 59.3±18.4 respectively, and the difference was statistically significant ( P<0.001). Subjective scores showed statistically significant differences among the 5 groups (χ2=101.61, P<0.001). The score of the 40 keV group was higher than that of the 60 keV group, 70 keV group, and conventional group ( P<0.001), and there was no significant difference when compared with the subjective score of the 50 keV group ( P>0.05). Conclusions:The 40 keV monochromatic image of dual detector spectral CT is the best image to observe the lumen of the stent after TIPS.