Objective:To establish a lung cancer model of patient-derived tumor xenografts (PDTX) and to explore the relation-ship between primary cisplatin resistance and ERCC1 and IGFBP5 expression levels. Methods:Lung cancer tissues from 84 patients who underwent surgery were collected and implanted into nude mice. Patient characteristics for the first generation xenografts that were and were not engrafted were compared. Passage 3 xenografts were treated with cisplatin. The expression levels of ERCC1 and IGFBP5 in cisplatin-resistant and cisplatin-sensitive groups were detected using immunohistochemistry assay. Results:The model success rates were 32.14%(27/84) in first-generation xenografts, 88.89%(24/27) in second-generation xenografts, and 95.83%(23/24) in third-gener-ation xenografts. The tumorigenicity of first-generation xenografts was correlated with size, differentiation, clinical stage, and histologi-cal type. PDTX tumors maintain the histological type of parental tumors through serial passage in nude mice. ERCC1 expression level was significantly higher in the cisplatin-resistant group than in the cisplatin-sensitive group, whereas the IGFBP5 expression level was lower in the cisplatin-resistant group than in the cisplatin-sensitive group. Conclusion:Lung cancer PDTX models were successfully es-tablished, and histological characteristics of the primary cancers were retained. Therefore, the models may serve a function in preclini-cal research of lung tumor biology and for exploring the drug resistance mechanism of tumors. The cisplatin resistance of primary lung cancer may be correlated with the expression level of ERCC1 and IGFBP5 in lung carcinoma.

Objective To discuss the technology of modest isolating and enriching the intestinal epithelial stem cells. Methods Mouse intestinal epithelial cells were stained by Rhodamine 123 (Rho), sorting the Rhodamine 123 low staining cell population ( Rholow ) and Rhodamine 123 strong straining cell population ( Rhobri ) by fluorescence activated cell sorting(FACS) in flow cytometer; Detecting the musaashi-1 and p-glycoprotein 1 (p-g-p1) mRNA expression of two groups by RT-PCR; Analyzing the cell cycle and the percentage of the musashi-1 positive cells by flow cytometry. Results The intestinal epithelial cells were divided into three groups, Rhodamine 123 low staining cells( 12. 34% of total cells), Rhodamine 123 middle staining cells (45.26% of total cells) and Rhodamine 123 strong staining cells ( 41. 40% of total cells). The Rholow cell fraction and Rhobri cell fraction were isolated successfully. Both of musashi-1 and p-g-p1 mRNA were strongly expressed in Rho1ow cell fraction, and Rhobri cell fraction little expressed p-g-p1 mRNA. Most of Rho1ow cells were in G0/G1 phase, and the musashi-1 positive cells were about 10.37% of total cells in this fraction. Conclusion The intestinal epithelial stem cells can be modestly isolated and enriched by Rhodamine 123 staining.

Infection is the most common complication and the most common cause of death in burn patients. It is very important to employ anti-infection measures reasonably and effectively for victims of major burns. However, a consensus of opinion of how to use systemic antibiotics in prophylaxis of infection in the early stage of burn is still lacking. The indications of the early systemic use of prophylactic antibiotics are discussed in this article.
Anti-Bacterial Agents ; therapeutic use ; Antibiotic Prophylaxis ; methods ; Burns ; complications ; drug therapy ; Humans ; Wound Infection ; chemically induced ; prevention & control

Objective To construct a recombinant fusion protein with pneumococcal surface pro-tein A (PspA) of Stretococcus pneumonia (SPN) familyⅠclade 1 and 2, and to analyze the immunogenici-ty of the fusion protein.Methods The gene fragments encoding theα-helix of PspA of the two clades were amplified by PCR and then inserted into the expression vector pET-27b(+) to construct the recombinant ex-pression plasmid.The transformed Escherichia coli BL21 strains carrying expression plasmid were induced by IPTG to express the recombinant protein.The titers and affinity of antibodies against PspA protein were measured by ELISA.An opsonophagocytic assay and an animal experiment were performed to evaluate the immunogenicity of the recombinant protein.Results Double enzyme cutting and gene sequencing confirmed the two purpose gene fragments were correctly expressed in the expression vector pET-27b(+).The titers of anti-PspA antibody in the serum of Kunming ( KM) mice immunized with the fusion protein were 1 ×104 . The affinity of anti-PspA antibody reached to 2×105 .The rates of recombinant PspA6B-PspA05 protein me-diated phagocytosis for SPN6B, SPN05 and SPN01 strains were 20%, 15% and 8.8%, respectively.No SPN23F strain was engulfed by macrophages upon the stimulation with PspA6B-PspA05 protein.The survival rates of mice injected with SPN05, SPN6B, SPN01 and SPN23F strains were respectively 75%, 92%, 75%and 33%upon the immunization of PspA6B-PspA05 protein.Conclusion The recombinant fusion protein PspA6B-PspA05, constructed with the PspA proteins of Stretococcus pneumonia familyⅠclade 1 and 2, was successfully expressed in the E.coli prokaryotic system with the advantage of high immunogenicity.High ti-ters of anti-PspA antibodies with high specificity were induced in KM mice upon the stimulation with Ps-pA6B-PspA05 protein.Moreover, a cross-protective immunity was induced in KM mice upon the immuniza-tion with PspA6B-PspA05 protein.

Objective To explore the inter-regional,base-like and informatized support of the field medical station during rotational training.Methods The field medical station information system developed by the hospital was introduced,which had the working mode involving in a set of system and two kinds of terminals.The problems of the information system were analyzed during iner-regional,base-like rotational training.Results The information system had its functions realized,and stills had to be improved in casualty information input flow,precision materials management and allocation standard of operating terminal.Conclusion The field medical station information system contributes to enhancing its service efficiency and informatization.

OBJECTIVETo explore the value of application of Bioflex dynamic stabilization system in treating multi-segment lumbar degenerative disease.

METHODSClinical datas of 13 patients with multi-segment lumbar degenerative disease (8 males and 5 females,ranging in age from 51 to 72 year with an average of 65.0) were retrospectively analyzed between April 2008 and May 2009. The involved area included L3-S1 in 7 cases, L2-S1 in 3 cases, L3-L5 in 1 cases, L4-S1 in 2 cases. All patients underwent decompression, dynamic stabilization with Bioflex system, according to the severity of degenerative disc with/without interbody fusion. The clinical effects were evaluated by VAS, ODI. ROM and fusion segments were also observed.

RESULTSThe mean follow up period was 19.5 months (from 12 to 26 months). The mean operative time was 183.4 min (from 90 to 240 min) and the mean volume of blood loss was 610.2 ml (from 400 to 1 220 ml). The mean VAS score was 7.8 +/- 1.3 preoperatively, 2.3 +/- 0.9 postoperatively and 2.1 +/- 0.8 at the last follow up. The average ODI was (60.50 +/- 4.40)% preoperatively, (17.80 +/- 2.10)% postoperatively and (16.20 + 2.40)% at the last follow up. The VAS and ODI significant improved in postoperatively (P < 0.05), and there was no statistical difference between postoperative and last follow up (P > 0.05). ROM of whole lumbar and non-fused segment showed obviously decreased and adjacent segment showed insignificant increased. The fusion rate of interbody fusion level was 95.0% (19/20).

CONCLUSIONThe preliminary clinical results show the Bioflex system combined with intebody fusion is a safe and effective technique in treating multi-segment lumbar degenerative disease.

Aged ; Female ; Humans ; Internal Fixators ; Intervertebral Disc Displacement ; surgery ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Retrospective Studies ; Spinal Fusion ; methods ; Spinal Stenosis ; surgery

This study was aimed to establish the PCR methods to detect nucleophosmin (NPM) gene and its mutation. 2 leukemia cell lines and 23 specimens from patients with acute myelogenous leukemia (AML) were investigated. The level of NPM mRNA was detected by RT-PCR. The exon-12 of NPM gene in leukemia cell lines was amplified by PCR and sequenced. Using the plasmid containing cDNA of NPM mutation A as a positive template, the PCR procedure to detect mutation A was established and evaluated. Then, the mutation of NPM was analyzed in 23 AML specimens. The results indicated that the expression level of NPM in leukemia cell lines was higher than that in normal cells. Different overexpression levels of NPM mRNA were found in all 23 AML specimens. PCR indicated that mutation had been not occurred at NPM exon-12 in THP1 and K562 cells, but a T base was deleted at 3' untranslated region of NPM gene in K562 cells. The PCR used for directly detecting NPM mutation A can specially amplify the NPM mutation gene. The method was reproducible, whose coefficient of variability was 1.6% and 3.1% in intra-and inter-assays respectively. The lowest detectable limit was 100 pg cDNA. Using the PCR methods, NPM mutation A could be detected in 2 out of 23 AML specimens, but NPM mutation A was not found in THP1 and K562 cells. It is concluded that the RT-PCT method detecting NPM mRNA level and the PCR method detecting directly NPM mutation are established. NPM mRNA is overexpressed in leukemia cells; NPM mutation A occurs in some AML patients.
Adult ; Base Sequence ; Female ; Humans ; Leukemia, Myeloid, Acute ; genetics ; Male ; Middle Aged ; Molecular Sequence Data ; Mutation ; Nuclear Proteins ; genetics ; Polymerase Chain Reaction ; methods ; RNA, Messenger ; genetics ; metabolism ; Young Adult

OBJECTIVETo study the mechanism of hepatitis B virus infected patients who is negative for HbsAg.

METHODSDNA sequences of 46 patients were analyzed. In these patients, HBsAg was negative but HBV DNA was positive and six new HBsAg variants were identified. Four of the six variants were combined point mutants and two were insertion variants. These S genes were subcloned into eukaryotic expression vector EBO-plpp, and the recombinant eukaryotic expression plasmids were transfected into COS7 cells. Cell lines expressing mutant type HBsAg were obtained. The supernatants were detected by ELISA and RIA.

RESULTSOnly the two-amino acid-insertion variants could be detected and the others failed to react with polyclonal and monoclonal antibodies against HbsAg.

CONCLUSIONThe results indicated that the point mutations and insertions may result in a conformational change of the S gene, which affect HBsAg antigenicity, suggesting a possible relationship between the variants and the negative conversion of HBsAg of the patients.

Animals ; Antigenic Variation ; COS Cells ; Cercopithecus aethiops ; Hepatitis B Surface Antigens ; genetics ; immunology ; Hepatitis B virus ; genetics ; immunology ; Hepatitis B, Chronic ; immunology ; virology ; Humans ; Plasmids ; genetics ; Point Mutation ; Transfection

Nucleophosmin (NPM) is a protein that shuttles between the nucleus, nucleoplasm and cytoplasm. NPM gene mutations and aberrant cytoplasmic NPM localization have been recently described in acute myelogenous leukemia (AML) with normal karyotype and in a few myelodysplastic syndromes. Expression of NPM mutant reduces the ability of Arf to initiate a p53 response and to induce cell cycle arrest. Clinical research has revealed that NPM mutations are relative to prognosis and can be used to monitor and quantify minimal residual disease (MRD) in AML patients with normal karyotype, therefore, these findings indicate that nucleophosmin mutations might contribute to illustration of myeloid leukemogenesis. In this paper, the research progress of nucleophosmin mutations in haematological malignancies was reviewed.
Cell Nucleolus ; metabolism ; Hematologic Neoplasms ; genetics ; pathology ; Humans ; Mutation ; Nuclear Proteins ; genetics ; fms-Like Tyrosine Kinase 3 ; genetics

OBJECTIVETo determine the changes of serum Tau protein, glial fibrillary acidic protein (GFAP), tumor necrosis factor alpha (TNF-α), and malonaldehyde (MDA) in rats after blast-related traumatic brain injury (BTBI) and to provide relative information for further studies on BTBI mechanism and seek specific biomarkers for BTBI.

METHODSNinety male Sprague-Dawley rats were randomly assigned into three groups: control group, moderate blast injury group, and severe blast injury group (n=30 for each). Rats in the moderate and severe blast injury groups were respectively exposed to corresponding levels of BTBI. After explosion, serum levels of Tau, GFAP, TNF-α, and MDA in each group were determined by Elisa assay at different time points after injury (8 h, 24 h, 3 d, and 6 d). The extent of brain damage was detected by Nissl staining and TUNEL assay.

RESULTSSerum levels of Tau and GFAP rapidly increased and reached the peak at 24 h after either moderate or severe blast injury. All the values were significantly higher than control group at all time points (P<0.05). Serum TNF-α level of both injury groups peaked at 8 h after BTBI and stayed significantly higher than control group at all time points (P<0.05). Serum MDA of two injury groups began to significantly increase at 3 d and the level stayed significantly higher than control group until 6 d (P<0.05). Moreover, unlike the other biomarkers, serum MDA of severe blast injury group was significantly higher than moderate blast injury group at 6 d (P<0.05).

CONCLUSIONThe changes of serum Tau, GFAP, and TNF-α showed a good sensitivity at the acute phase after BTBI (within 24 h). However, their specificity and correlation with the extent of injury were limited in this experiment. Moreover, although the change of serum MDA showed a poor sensitivity and specificity to the diagnosis of BTBI during the first few days, it can reflect the injury degree at 6 d after injury. Therefore, further studies are needed to improve the methods of detecting more serum markers and investigate the significance of multiple markers in diagnosing BTBI.

Animals ; Biomarkers ; blood ; Blast Injuries ; blood ; Brain Injuries ; blood ; Enzyme-Linked Immunosorbent Assay ; Glial Fibrillary Acidic Protein ; blood ; Male ; Malondialdehyde ; blood ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; analysis ; tau Proteins ; blood