OBJECTIVETo investigate the appropriate extubation time and treatment of late complications after early tracheotomy in patients with moderate or severe inhalation injury.

METHODSOne hundred and fifty patients (105 males and 45 females) with inhalation injury were admitted to our hospital from January 2000 to January 2009. Among them, 109 out of 129 cases with moderate inhalation injury received early tracheotomy, and all 21 cases with severe inhalation injury received early tracheotomy. Data were collected for analysis as follows: (1) incidence of re-intubation due to suffocation and pneumonia incidence after extubation within 2 weeks or after 2 weeks post inhalation injury (PII), and mortality rate within the first week after injury were recorded. (2) Conservative treatments including expectorant, oral antibiotics, and absolute bedrest were recommended for patients who had severe cough, hoarseness or poor pulmonary function after late extubation and closure of tracheostomy wound. Fiberoptic bronchoscopy findings (tracheostenosis degree, granuloma formation rate, vocal cord paralysis rate) and pulmonary function index (FEV(1)) data were collected and analyzed in 30 cases with moderate inhalation injury and 10 cases with severe inhalation injury within 3 months after injury for follow-up. Data were processed with t test or chi-square test.

RESULTSThere was no obvious difference in the rate of re-intubation after extubation in patients with moderate inhalation injury between those done within 2 weeks PII (15/70, 21.4%) and those done after 2 weeks PII (2/25, 8.0%) (χ(2) = 1.52, P > 0.05). Pneumonia incidence in patients of moderate inhalation injury with extubation within 2 weeks PII (21/70, 30.0%) was lower than those with extubation after 2 weeks PII (15/25, 60.0%) (χ(2) = 7.04, P < 0.05). Levels of above-mentioned indexes in patients with severe inhalation injury extubated in different stages were similar to those of patients with moderate inhalation injury. Within the first week after injury, mortality rate of patients with severe inhalation injury was higher than that of patients with moderate inhalation injury (χ(2) = 11.90, P < 0.05). During follow-up, tracheostenosis rate in patients with moderate or severe inhalation injury was 100.0%; granuloma formation rate and vocal cord paralysis rate in patients with severe inhalation injury were higher than those of patients with moderate inhalation injury (with χ(2) value respectively 4.59, 13.47, P values all below 0.05). The FEV(1) value of patients with moderate inhalation injury in the 1st, 2nd, 3rd month after injury was respectively higher than that of patients with severe inhalation injury (with t value respectively 5.48, 12.10, 6.25, P values all below 0.05). The values recovered to normal level in the 3rd month after injury.

CONCLUSIONSExtubation time of tracheotomy for patients with moderate or severe inhalation injury within 2 weeks or after 2 weeks PII has its own advantage and disadvantage, and it should be performed according to specific conditions of each patient. Conservative treatment is optional for late complications of respiratory system.

Adolescent ; Adult ; Aged ; Burns, Inhalation ; surgery ; Child ; Child, Preschool ; Female ; Follow-Up Studies ; Humans ; Intubation, Intratracheal ; adverse effects ; Male ; Middle Aged ; Postoperative Complications ; Tracheotomy ; adverse effects ; Young Adult

To elucidate the effect of established primary bone marrow stromal layers on the gene transduction of human hematopoietic stem/progenitor cells (HSC/HPC), mononuclear cells (MNC) from adult bone marrow were isolated by centrifugation on Ficoll-Hypaque gradients and plated in stromal culture medium. The cells were incubated until passage 4 to establish primary stromal layers. The HSC/HPC prestimulated by cytokines were transduced by retroviral supernatant containing mdr1 gene in presence of irradiated stroma-contact support. Transduced cells were plated in a colony-forming unit assay with and without vincristine (VCR) to assess the efficiency of transduction. Individual colonies were also analyzed by polymerase chain reaction (PCR) for the presence of provirus. The results showed that the mixed adherent cell layers were formed when adult bone marrow stromal cells were incubated for four to six weeks, mainly being composed of fibroblasts. In the presence of stroma-contact support, the average of gene transduction efficiency in marrow-derived progenitors increased 2.1 to 3.3 folds measured by colony-forming assay and/or PCR, significantly higher than those without support of stroma. It is concluded that the presence of bone marrow stroma support in combination with cytokine facilitates augmenting the extent of retroviral-mediated gene transduction.
Bone Marrow Cells ; physiology ; Genes, MDR ; Hematopoietic Stem Cells ; metabolism ; Humans ; Retroviridae ; genetics ; Stromal Cells ; physiology ; Transduction, Genetic

OBJECTIVETo study the chemical constituents of Serissa serissoides.

METHODChemical constituents were isolated with the column chromatographic methods including silica gel and sephadex LH -20, and their structures were elucidated on the basis of their spectral and chemical evidences.

RESULTEight compounds were obtained and were identified as: palmitic acid (1), corosolic acid (2), daucosterol (3), urs-12-en-28-ol (4), oleanolic acid (5), uroslic acid (6), 4-hydroxy-3-methoxybenzoic acid (7), and 2,6-dimethoxy-p-benzoquinone (8).

CONCLUSIONExcept compound 5, other seven compounds were found from the genus Serissa for the first time.

Chromatography, Gel ; methods ; Palmitic Acid ; chemistry ; isolation & purification ; Plants, Medicinal ; chemistry ; Rubiaceae ; chemistry ; Sitosterols ; chemistry ; isolation & purification ; Triterpenes ; chemistry ; isolation & purification

OBJECTIVETo establish a novel human monocytic leukemic cell line and characterize its biological features.

METHODSMononuclear cells isolated from the bone marrow of an acute monocytic leukemia (AML-M(5b)) patient at relapse were inoculated in a liquid culture system. And the biologic features of the established cell line SHI-1 were characterized by morphology, cytochemical staining, flow cytometry, karyotypic analysis, polymerase chain reaction (PCR), reverse transcriptase PCR (RT-PCR), fluorescence in situ hybridization (FISH), tumorigenicity in nude mice, quantitative fluorescent polymerase chain reaction, broth medium culture, short tandem repeating sequences-PCR (STR-PCR), multiplex-FISH (M-FISH), and (3)H-thymidine incorporation assay.

RESULTSA human acute monocytic leukemia cell line, SHI-1, was established and has proliferated continuously in vitro for over one year. The cell line presented typical morphology and immuno-profile of monocytic lineage with the original t(6;11)(q27;q23) and del(17)(p11) abnormalities. The MLL-AF6 fusion transcript was detected by RT-PCR. The rearrangement of MLL gene, deletion of p53 gene, and translocation between chromosomes 6 and 11 were revealed by FISH. A point mutation of ATC-->ACC at exon 6 of the p53 gene was found by sequencing of the PCR products. The clonality and the high tumorigenicity of the SHI-1 cell line were confirmed. Infections of EBV and mycoplasma were excluded. A derivative chromosome 7 resulting from a translocation between chromosomes 7 and 13, monosomy 18 and a minute derived from chromosome 8 in addition to t(6;11) and deletion(17)(p11) were detected by M-FISH in SHI-1 cells passaged to March 2003. Cell line authentication by STR-PCR confirmed the identity to the original leukemic cells of the patient. (3)H-thymidine incorporation assay showed that IL-4 and IL-15 had proliferative effects, while IFN-gamma, TNFalpha, IL-2, PDGF, and IL-7 had inhibitory effects on the cell line.

CONCLUSIONSSHI-1 is a novel acute monocytic leukemia-derived cell line carrying t(6;11)(q27;q23) and p53 gene alteration and having high tumorigenicity in nude mice. It provides a new useful tool for leukemia research.

Adult ; Animals ; Bone Marrow Cells ; metabolism ; pathology ; Cell Line, Tumor ; Chromosomes, Human, Pair 11 ; genetics ; Chromosomes, Human, Pair 6 ; genetics ; Female ; Humans ; In Situ Hybridization, Fluorescence ; Karyotyping ; Leukemia, Experimental ; blood ; genetics ; pathology ; Leukemia, Monocytic, Acute ; blood ; genetics ; pathology ; Male ; Mice ; Mice, Nude ; Translocation, Genetic ; Transplantation, Heterologous ; Tumor Suppressor Protein p53 ; genetics

OBJECTIVETo investigate the feasibility of reproduce hypertrophic scar and keloid in nude mice in the study of pathological scars.

METHODSPieces (0.8 x 0.8 x 0.5 cm) of hypertrophic scars and keloids were implanted into subcutaneous tissue of the nude mice for 16 days, during this period the gross condition of the nude mice and the state of the implants were observed. The implants were extracted after 16 days, and the volume, the microscopic characteristics of the scar, the content of acid mucopolysaccharide, and different types of collagen were determined and compared with that of the original specimens.

RESULTSAll mice survived with nice wound healing after the surgery. There was no obvious difference in the acid mucopolysaccharide content in keloid and hyperplastic scar before implantation (3448 +/- 1452, 1940 +/- 509), and after implantation (3237 +/- 1871, 1809 +/- 552, P > 0.05). The implants maintained the collagen pattern, with no signs of cell degeneration and necrosis.

CONCLUSIONThis experiment showed that the viability and morphology of hypertrophic scars and keloids were maintained after they were implanted in nude mice. Therefore it is feasible to use nude mice as the animal model in the study of hypertrophic scars and keloids.

Animals ; Cicatrix, Hypertrophic ; pathology ; Disease Models, Animal ; Keloid ; pathology ; Mice ; Mice, Inbred BALB C ; Mice, Nude

OBJECTIVETo explore early diagnosis of hemophagocytic syndrome (HPS) and effective treatment.

METHODSA multicenter retrospective study was carried out to analyze the causes, clinical features, laboratory findings, treatment and clinical outcomes of 72 patients with HPS.

RESULTSAmong the 72 patients, EBV infection and T lymphoma were the most common initiating diseases. The most common clinical features were persistent fever (100%) and splenomegaly (83.3%). The diagnostic sensitivity was persistent fever (100%), peripheral cytopenia in two or more lineages (97.2%), high concentration of serum soluble CD25 (93.1%) and low NK cell activity (94.4%). The median percentage of serum glycosylated ferritin was significantly lower in patients in HPS group \[(17.4 +/- 16.0)%\] than in control group \[(53.6 +/- 13.3)%\] (P < 0.01). And the median level of serum TNF-alpha was significantly higher in patients group \[(143.2 +/- 64.8) microg/L\] than in controls \[(66.9 +/- 19.4) microg/L\] (P < 0.01). Hepatic dysfunction was seen in most patients (83.6%) mainly manifested as elevated liver enzymes and hypoalbuminemia. The 15-week total survival rate was 46.8% in 47 treated patients, and was 63% in 27 treated with fludarabine in combination with high dose methylprednisolone. The platelet count and fibrinogen level were significantly lower in death group than in survival group.

CONCLUSIONSThe diagnostic sensitivities of presistent fever, peripheral cytopenia in two or more lineages, high concentration of serum soluble CD25 and low NK cell activity are relatively high and lacking hemophagocytosis does not exclude the diagnosis. Low percentage of glycosylated ferritin and high concentration of TNF-alpha would be helpful to the diagnosis. High dose methylprednisolone combined with fludarabine is an effective therapy. Platelet count and fibrinogen level are poor prognostic factors for HPS.

Humans ; Lymphohistiocytosis, Hemophagocytic ; diagnosis ; Methylprednisolone ; Retrospective Studies ; Treatment Outcome ; Tumor Necrosis Factor-alpha

This study was purposed to explore the tumorigenicity of a novel human monocytic leukemic cell line SHI-1 in nude mice and its mechinism. The tumorigenicity in mice was evaluated in sixteen nude mice subcutaneously injected with the SHI-1 cell line. The tumor specimen was studied by the conventional pathologic examination. The mononuclear cells (MNC) of the tumor was assayed by RHG banding, the transcription of MLL-AF6 fusion gene and the VEGF gene was detected by RT-PCR. Gelatin zymography method was used to study the expression of MMP-9 and MMP-2 in the supernatant of the SHI-1 cell line. Matrigel invasion assay was employed for the study of migration of the SHI-1 cell in vitro. The results showed that the tumor masses were found in all sixteen mude mice after subcutaneous injection of SHI-1 cells, the tumor mass was mainly composed of leukemia cells, the transcription of MLL-AF6 fusion gene and VEGF gene was proved by RT-PCR analysis, the expressions of MMP-2 and MMP-9 in the serum-free culture supernatant of the SHI-1 cell line were significantly higher than those in U937, K562, and NB4 cell lines. The SHI-1 cell line exhibited significantly higher in vitro invasiveness than other leukemia cell lines, the blocking antibody of MMP-2 could inhibit the migration of the SHI-1 cell line significantly. It is concluded that the SHI-1 cell line presents higher tumorigenicity in nude mice than other leukemia cell line and the mechanism is associated with p53 gene alteration, high transcription level of VEGF gene, high expression level of MMP, and significantly higher invasiveness.
Animals ; Disease Models, Animal ; Genes, p53 ; genetics ; Humans ; Leukemia, Monocytic, Acute ; pathology ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Tumor Cells, Cultured ; Vascular Endothelial Growth Factor A ; genetics

OBJECTIVETo identify the HBV genotype-specific tag sequence.

METHODSThe large S region sequences from 930 HBV genomes were aligned to identify the genotype-specific tag sequences. PCR was used to check the genotyping effect of these tags.

RESULTSTwo tag sequences, sequence between 149-169 and sequence between 461-483, were identified in the large S region. Using primers specific to these tag sequences, the genotype of HBV can be specifically identified.

CONCLUSIONThese tag sequences can be used for HBV genotyping.

Base Sequence ; DNA Primers ; Gene Library ; Genes, Viral ; Genotype ; Hepatitis B virus ; classification ; genetics ; Hepatitis B, Chronic ; virology ; Humans ; Molecular Sequence Data ; Polymerase Chain Reaction ; methods ; Protein Precursors ; genetics ; Sequence Analysis, DNA

The identification of genes inducing resistance to anticancer chemotherapeutic agents and their introduction into hematopoietic cells represents a promising approach to overcome bone marrow toxicity, the limiting factor for most high-dose chemotherapy regimens. Because resistance to cyclophosphamide has been correlated with increased levels of expression of the aldehyde-dehydrogenase (ALDH1) gene in tumor cells lines in vitro, this study tested whether ALDH1 overexpression could directly induce cyclophosphamide resistance. Results showed that a retroviral vector was used to transduce full-length human ALDH1 cDNA into human hematopoietic cell line K562 that was then tested for resistance to 4-hydroxycyclophosphamide (4-HC), an active analogue of cyclophosphamide. Overexpression of the ALDH1 gene resulted in a significant increases in cyclophosphamide resistance in transduced K562 cells (50% inhibition concentration, IC50 = 10 micro mol/L). These findings indicate that ALDH1 overexpression is sufficient to induce cyclophosphamide resistance in vitro and provide a basis for testing the efficacy of ALDH1 gene transduction to protect bone marrow cells from high-dose cyclophosphamide in vivo.
Aldehyde Dehydrogenase ; genetics ; Antineoplastic Agents, Alkylating ; pharmacology ; Cell Division ; drug effects ; Cell Survival ; drug effects ; Cyclophosphamide ; analogs & derivatives ; pharmacology ; Dose-Response Relationship, Drug ; Drug Resistance, Neoplasm ; Gene Expression Regulation, Enzymologic ; Genetic Vectors ; genetics ; Humans ; Inhibitory Concentration 50 ; K562 Cells ; drug effects ; enzymology ; metabolism ; Retroviridae ; genetics ; Transfection

The study was aimed to investigate the incidences and risk factors of acute and chronic graft-versus-host diseases (GVHD) and to clarify their effects on relapse and survival of recipients after allogeneic hematopoietic stem cell transplantation (allo-HSCT). The clinical data of 100 cases of allo-HSCT were retrospectively analyzed. The incidences and risk factors of aGVHD and cGVHD, relapse and survival were studied. The results showed that 31 cases developed aGVHD of II - IV grade (34.4%) and 14 cases developed aGVHD of III - IV grade (17.7%). HLA matched or mismatched did not show significant difference in the development of aGVHD of II - IV grade (p > 0.05). Previous occurrence of aGVHD was the risk factor for cGVHD (HR = 2.303, p = 0.088). The female was a favorable factor for cGVHD (HR = 0.401, p = 0.055). The relapse rate was lower in patients who developed cGVHD. The development of aGVHD of II - IV grade was the risk factor for overall survival (p < 0.05). The mortality of patients with aGVHD of III - IV grade and mortality of patients with aGVHD of 0 - I grade were 81.0% and 35.7% respectively, there was very significant difference between them (p = 0.000). In conclusion, till now GVHD and graft-versus-leukemia (GVL) effect can not be separated. The positive effect of GVL could be counteracted by GVHD-related mortality. It is necessary to prevent and control the development of severe aGVHD. The development of local cGVHD may be beneficial to the long-term disease-free survival of patients after allo-HSCT.
Adolescent ; Adult ; Child ; Child, Preschool ; Disease-Free Survival ; Female ; Graft vs Host Disease ; etiology ; mortality ; Hematopoietic Stem Cell Transplantation ; adverse effects ; mortality ; Humans ; Incidence ; Male ; Middle Aged ; Recurrence ; Retrospective Studies ; Risk Factors ; Young Adult