OBJECTIVETo construct the cloning vector of glycoprotein G2 gene of hantavirus (HV), to analyze the sequence of G2 gene by the phylogenetic tree, and to study the differences among glycoprotein G2 genes from the world around.

METHODSEnvelope glycoprotein G2 gene was amplified from four specimens of Shandong province by RT-PCR, and the product recombined into the PMD-18T vector. The clones that carry the G2 gene were identified. After sequencing, the gene sequence was handled with the software DNASTAR, compared with 24 strains worldwide and the phylogenetic tree was drawn.

RESULTSHV G2 gene was amplified by RT-PCR from 4 specimens, named GM04-38.G2, ZB8.G2, JUN5-14.G2, RCH5.G2, respectively. The map of the phylogenetic tree showed that all the 4 strains belonged to SEO-type hantavirus. The analysis of the sequence showed that all the four HV strains had the highest rates of homology with Z37 strain. The sequence homology of SEO-type HV strains was from 82.3% to 99.8%.

CONCLUSIONThe four cloning vectors containing the glycoprotein G2 genes were successfully constructed. Envelope glycoprotein G2 gene of four specimens from Shandong province had high homology rates.

Animals ; China ; Cloning, Molecular ; Hantavirus ; genetics ; Mice ; Phylogeny ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, DNA ; Viral Envelope Proteins ; genetics

The study is aimed to ensure the quality and safety of medicinal plants by using ITS2 DNA barcode technology to identify Corydalis boweri, Meconopsis horridula and their close related species. The DNA of 13 herb samples including C. boweri and M. horridula from Lhasa of Tibet was extracted, ITS PCR were amplified and sequenced. Both assembled and web downloaded 71 ITS2 sequences were removed of 5. 8S and 28S. Multiple sequence alignment was completed and the intraspecific and interspecific genetic distances were calculated by MEGA 5.0, while the neighbor-joining phylogenetic trees were constructed. We also predicted the ITS2 secondary structure of C. boweri, M. horridula and their close related species. The results showed that ITS2 as DNA barcode was able to identify C. boweri, M. horridula as well as well as their close related species effectively. The established based on ITS2 barcode method provides the regular and safe detection technology for identification of C. boweri, M. horridula and their close related species, adulterants and counterfeits, in order to ensure their quality control, safe medication, reasonable development and utilization.
Base Sequence ; China ; Corydalis ; chemistry ; classification ; genetics ; DNA Barcoding, Taxonomic ; methods ; DNA, Plant ; chemistry ; genetics ; DNA, Ribosomal Spacer ; chemistry ; genetics ; Molecular Sequence Data ; Nucleic Acid Conformation ; Papaveraceae ; chemistry ; classification ; genetics ; Phylogeny ; Plants, Medicinal ; chemistry ; classification ; genetics

Objective To evaluate the feasibility, safety and efficacy of intra-arterial thrombolytic therapy on elderly patients (≥ 80 years old) with acute ischemic stroke. Methods The clinical data of 86 patients with acute ischemic stroke, received intra-arterial thrombolytic therapy, were retrospectively analyzed; according to age differences, these patients were divided into advanced age group (≥80 years old, n=21) and common age group (<80 years old, n=65); and control group (≥80 years old, not receiving thrombolytic therapy, n=50) was established. The recanalization rate and early clinical improvement rate, and the incidence, recover rate and death rate of symptomatic intracerebral hemorrhage were evaluated in these patients after treatment. Results No significant differences in the favorite recanalization rate and short-term outcome, and the incidence of symptom intracranial hemorrhage were noted between the advanced age group and common age group (P=0.528, P=0.102,P=0.353). The incidence of symptom intracranial hemorrhage in the advanced age group was obviously higher than that in the control group (P=0.034); the recover rate of symptom ntracranial hemorrhage in the advanced age group (42.9%) was obviously lower than that in the common age group (50.8%), but significantly higher than that in the control group (16%, P=0.042, P=0.017). The mortality of the advanced age group was similar to that of the control group (23.8% versus 28%, P=0.816), but higher than that of common age group (23.8% versus 10.8%, P=0.034). Conclusion Relatively high feasibility, safety and efficacy of intra-arterial thrombolytic therapy are noted in elderly patients (≥80 years old) with acute ischemic stroke, demonstrating that the use of intra-arterial thrombolytic therapy in very elderly patients should not be avoided but pursued advisably.

Objective To investigate the effect of all-trans retinoic acid (ATRA) on expressions of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) in glioma stem cells (GSCs). Methods GSCs were isolated from human glioblastoma cell line U87 and identified by detecting the expressions of CD133 and nestin with immunofluorescence staining. The obtained GSCs were divided into control group,empty vector group (cultured with dimethyl sulfoxide [DMSO]) and ATRA treatment group (cultured with 10 nmool/L ATRA).After 10 d of differentiation; the proliferation of the treated GSCs was evaluated using CCK8 assay; the expressions of glial fibrillary acidic protein (GFAP),β-tubulin Ⅲ and galactoeerebroside (GralC) in the cells were detected by immunofluorescence.VEGF and bFGF levels in cultured supernatant were measured by ELISA; the mRNA expressions of VEGF and bFGF were detected by RT-PCR. Results The target antibodies of neural stem cells (NSCs), CD133 and nestin,positively expressed in the GSCs; differentiated GSCs can differentiate several kinds of homologous daughter cells,which expressed the cell markers of astrocytes,neurons and oligodendrocytes: GFAP, β-tubulin LⅢ and GalC, respectively. The percentage of GFAP-positive differentiated GSC s in the ATRA treatment group was significantly higher as compared with that in the other 2 groups after 10 d of differentiation (P＜0.05); the speed of proliferation of GSCs in ATRA treatment group was obviously slower than that in the other 2 groups 3-7 d after differentiation (P＜0.05).The VEGF and bFGF levels and the mRNA expression levels of VEGF and bFGF in GSCs of the ATRA treatment group 24 h after differentiation were also significantly lower than those in the other 2 groups (P＜0.05). Conclusion ATRA can induce the differentiation of GSCs and inhibit its proliferation.It may exerts its anti-glioblastoma effect through the VEGF and bFGF signaling pathways.

OBJECTIVETo explore the clinical and laboratory features and the prognosis of juvenile dermatomyositis (JDM) complicated with interstitial lung disease (ILD).

METHODData of 39 cases of JDM complicated with ILD hospitalized in Beijing Children's Hospital from January 2005 to December 2011 were collected. The clinical features, laboratory data and prognosis of these children were analyzed.

RESULTOf the 39 cases studied, 16 were boys, and 23 girls. The average age of onset was 5.6 years, and 61.5% of the patients' age of onset (24 cases) was under 6 years. Rashes (17 cases, 43.6%), simultaneous eruption of rashes and muscle weakness (14 cases, 35.9%), fever (4 cases, 10.1%), or muscle weakness (3 cases, 7.7%) were common initial symptoms of the disease. Only 51.3% of the patients (20 cases) had the symptoms of respiratory system, but (24 cases) 61.5% were complicated with that of the gastrointestinal system; (27 cases) 69.2% had at the same time electrocardiographic and echocardiographic abnormalities. The chest high resolution computed tomography (HRCT) showed cord or band-like shadows in their lungs of more than half of the cases (25 cases, 64.1%), and other changes included ground glass-like shadow (10 cases, 25.6%), net and lineation-like shadow (9 cases, 23.1%), nodular change (5 cases, 12.8%). The patients complicated with lung essential infiltration accounted for as high as 71.8% (28 cases). These imaging changes were largely seen on both dorsal sides of their lungs. Severe patients also had mediastinal emphysema, pneumothorax, pneumorrhagia or aerodermectasia. Twenty-four patients underwent pulmonary function examination, and 62.5% of the patients' pulmonary function (15 cases) was abnormal. The fatality rate of the cases studied was 10.1%.

CONCLUSIONThe imaging changes of patients suffering from JDM with ILD were often more severe as compared to the clinical symptoms, and were often complicated with damages to other systems and organs. The prognosis of those patients was poorer than others. Patients with JDM especially at a younger age of onset and with various organ damages should be examined with chest HRCT examinations as early as possible.

Child ; Child, Preschool ; Dermatomyositis ; complications ; diagnosis ; drug therapy ; Female ; Glucocorticoids ; administration & dosage ; therapeutic use ; Humans ; Immunosuppressive Agents ; administration & dosage ; therapeutic use ; Lung ; diagnostic imaging ; pathology ; Lung Diseases, Interstitial ; diagnosis ; drug therapy ; etiology ; Male ; Methotrexate ; administration & dosage ; therapeutic use ; Muscle Weakness ; diagnosis ; epidemiology ; etiology ; Prognosis ; Respiratory Function Tests ; Retrospective Studies ; Tomography, X-Ray Computed

OBJECTIVETo sum up the experience in treating very severe traumatic brain injuries.

METHODSRetrospective analysis of 68 patients with very severe traumatic brain injuries treated in our hospital from 1997 to 2002 was done.

RESULTSForty-one (60%) patients died. In the 50 patients treated surgically 27 (40%) survived, 8 recovered well, 9 had moderate disability and 10 had severe deficits. The 18 patients treated non-operatively all died.

CONCLUSIONSMuch attention should be given to the observation of the changes of severe brain injuries with cranial base injury. Timely operative decompression, basic life support, keeping effective brain blood perfusion and effective oxygen supply, improving cerebral microcirculation and preventing or controlling complications are the main methods to raise the successful rate of treating very severe brain injuries and the life quality of the patients.

Adolescent ; Adult ; Aged ; Brain Injuries ; diagnosis ; mortality ; therapy ; Cohort Studies ; Combined Modality Therapy ; Craniotomy ; Critical Care ; methods ; Decompression, Surgical ; Drug Therapy, Combination ; Female ; Glasgow Coma Scale ; Humans ; Injury Severity Score ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Oxygen ; therapeutic use ; Prognosis ; Retrospective Studies ; Risk Assessment ; Survival Analysis ; Treatment Outcome

Acute Disease ; Adult ; Aged ; Aneurysm, Dissecting ; etiology ; Female ; Humans ; Male ; Middle Aged ; Stents ; Subclavian Steal Syndrome ; complications ; diagnosis ; therapy

OBJECTIVETo investigate the influence of CD4+ CD25+ regulatory T cells(Treg cells) on mouse gastric cancer.

METHODSTreg cell in mouse spleen bearing gastric tumor was tested in different time points. Magic cell sorting(MACS) method was used to purify mouse Treg cells and the Treg cells were injected into mouse bearing gastric tumor with different dosage. After 3 weeks, the tumor size and tumor cell apoptosis rate were measured.

RESULTSTreg existed in normal mouse spleen with a rate of (3.86+/-0.07)%. In tumor model this percentage increased gradually and was (4.12+/-0.13)% after 3 weeks, which was significantly higher than that in control. When Treg cell applied in mouse reached 2.0 x 10(5), the tumor size enlarged significantly(P=0.013) and tumor cell apoptosis rate decreased significantly (P=0.012).

CONCLUSIONSTreg cell is associated with gastric cancer progress in mouse tumor model. Treg cell can promote gastric cancer growth and decrease tumor apoptosis. The anti- Treg GITR can improve anti- tumor effects.

Animals ; Apoptosis ; Female ; Flow Cytometry ; Male ; Mice ; Mice, Inbred Strains ; Spleen ; cytology ; Stomach Neoplasms ; immunology ; pathology ; T-Lymphocytes, Regulatory ; immunology

In order to reveal the cause of disease occurred in the process of Coptis chinensis growth, this paper studied the bacterial species diversity index of different aged rhizospheric and non-rhizospheric soil planting normal or sick C. chinensis by using PCR-DGGE technique. The representative DGGE bands were chosen to be cloned, and sequenced, the phylogeny were constructed. The results showed that the bacterial communities were very different between the normal and diseased soil samples of C. chinensis, and the diversity index (H) of diseased soil samples were higher than that of normal soil samples. Sequencing analysis of representative cloned DGGE bands showed that the unculturable bacteria were the dominant groups, and bacteria belonged to genus Bacillus, Acidovorax, Acinetobacter, uncultured Kluyvera, and uncultured Comamonas were also existing, but the reported plant pathogenic bacteria were not found in the C. chinensis planting soil. The density and brightness of clone band d in diseased soil samples was higher than that in normal soil sample, and sequencing analysis showed that it belonged to genus Acidovorax. Obviously, during the process of C. chinensis growth, the rhizospheric bacteria population changed, and the quantity of bacteria belong Acidovorax increased, which probably resulted in the disease occurred during C. chinensis growth.
Bacteria ; classification ; genetics ; isolation & purification ; Biodiversity ; Coptis ; growth & development ; microbiology ; Denaturing Gradient Gel Electrophoresis ; Molecular Sequence Data ; Phylogeny ; Polymerase Chain Reaction ; Rhizosphere ; Soil Microbiology

OBJECTIVETo investigate the possibility of differentiating prostate cancer (PCa) from benign prostatic disease by total prostate specific antigen (T-PSA) dynamic profiles following transrectal prostate biopsy, and to determine the cutoff value of the T-PSA ratio between pre- and post-biopsy.

METHODSA total of 36 men at the mean age of 69.89 years with increased serum PSA underwent prostate biopsy guided by transrectal ultrasound, followed by measurement of T-PSA at 10, 30, 60 and 90 min, plotting of T-PSA dynamic profiles and calculation of the pre- and post-biopsy T-PSA ratio at different time points. The patients were divided into a PCa and a non-PCa group according to the pathological results and compared for the difference in T-PSA ratios. The cutoff value of the pre- and post-biopsy T-PSA ratio was determined for the differentiation of PCa from benign prostatic diseases.

RESULTSThe post-biopsy T-PSA ratio was obviously higher in the non-PCa than in the PCa group (P < 0.05). With the ROC curve applied, the cutoff value of the T-PSA ratio was 1.5 and the best time for blood sampling was 30 minutes after the biopsy, with a 75% sensitivity and a 93% specificity.

CONCLUSIONEvaluation of the T-PSA ratio 30 minutes after biopsy might help screen the high-risk PCa population. Biopsy should be repeated for those with a lower T-PSA ratio in spite of initial benign results. The results are to be further supported by more prospective studies.

Aged ; Aged, 80 and over ; Biopsy, Needle ; methods ; Diagnosis, Differential ; Humans ; Male ; Middle Aged ; Prostate ; pathology ; Prostate-Specific Antigen ; blood ; Prostatic Diseases ; blood ; diagnosis ; pathology ; Prostatic Neoplasms ; blood ; diagnosis ; pathology ; Sensitivity and Specificity