Comparison of the upper and lower airway microbiome in early postoperative lung transplant recipients.

The upper and lower respiratory tract may share microbiome because they are directly continuous, and the nasal microbiome contributes partially to the composition of the lung microbiome. But little is known about the upper and lower airway microbiome of early postoperative lung transplant recipients (LTRs). Using 16S rRNA gene sequencing, we compared paired nasal swab (NS) and bronchoalveolar lavage fluid (BALF) microbiome from 17 early postoperative LTRs. The microbiome between the two compartments were significantly different in Shannon diversity and beta diversity. Four and eight core NS-associated and BALF-associated microbiome were identified, respectively. NS samples harbored more Corynebacterium, Acinetobacter, and Pseudomonas, while BALF contained more Ralstonia, Stenotrophomonas, Enterococcus, and Pedobacter. The within-subject dissimilarity was higher than the between-subject dissimilarity, indicating a greater impact of sampling sites than sampling individuals on microbial difference. There were both difference and homogeneity between NS and BALF microbiome in early postoperative LTRs. High levels of pathogens were detected in both samples, suggesting that both of them can reflect the diseases characteristics of transplanted lung. The differences between upper and lower airway microbiome mainly come from sampling sites instead of sampling individuals. IMPORTANCE: Lung transplantation is the only therapeutic option for patients with end-stage lung disease, but its outcome is much worse than other solid organ transplants. Little is known about the NS and BALF microbiome of early postoperative LTRs. Here, we compared paired samples of the nasal and lung microbiome from 17 early postoperative LTRs and showed both difference and homogeneity between the two samples. Most of the "core" microbiome in both NS and BALF samples were recognized respiratory pathogens, suggesting that both samples can reflect the diseases characteristics of transplanted lung. We also found that the differences between upper and lower airway microbiome in early postoperative LTRs mainly come from sampling sites instead of sampling individuals.

The Airway Microbiota Signatures of Infection and Rejection in Lung Transplant Recipients.

Infection and rejection are the two most common complications after lung transplantation (LT) and are associated with increased morbidity and mortality. We aimed to examine the association between the airway microbiota and infection and rejection in lung transplant recipients (LTRs). Here, we collected 181 sputum samples (event-free, n = 47; infection, n = 103; rejection, n = 31) from 59 LTRs, and performed 16S rRNA gene sequencing to analyze the airway microbiota. A significantly different airway microbiota was observed among event-free, infection and rejection recipients, including microbial diversity and community composition. Nineteen differential taxa were identified by linear discriminant analysis (LDA) effect size (LEfSe), with 6 bacterial genera, Actinomyces, Rothia, Abiotrophia, Neisseria, Prevotella, and Leptotrichia enriched in LTRs with rejection. Random forest analyses indicated that the combination of the 6 genera and procalcitonin (PCT) and T-lymphocyte levels showed area under the curve (AUC) values of 0.898, 0.919 and 0.895 to differentiate between event-free and infection recipients, event-free and rejection recipients, and infection and rejection recipients, respectively. In conclusion, our study compared the airway microbiota between LTRs with infection and acute rejection. The airway microbiota, especially combined with PCT and T-lymphocyte levels, showed satisfactory predictive efficiency in discriminating among clinically stable recipients and those with infection and acute rejection, suggesting that the airway microbiota can be a potential indicator to differentiate between infection and acute rejection after LT. IMPORTANCE Survival after LT is limited compared with other solid organ transplantations mainly due to infection- and rejection-related complications. Differentiating infection from rejection is one of the most important challenges to face after LT. Recently, the airway microbiota has been reported to be associated with either infection or rejection of LTRs. However, fewer studies have investigated the relationship between airway microbiota together with infection and rejection of LTRs. Here, we conducted an airway microbial study of LTRs and analyzed the airway microbiota together with infection, acute rejection, and clinically stable recipients. We found different airway microbiota between infection and acute rejection and identify several genera associated with each outcome and constructed a model that incorporates airway microbiota and clinical parameters to predict outcome. This study highlighted that the airway microbiota was a potential indicator to differentiate between infection and acute rejection after LT.

Sleep quality, anxiety and depression in advanced lung cancer: patients and caregivers.

OBJECTIVE: To investigate the clinical implications of sleep quality, anxiety and depression in patients with advanced lung cancer (LC) and their family caregivers (FCs). METHODS: A total of 98 patients with advanced LC and their FCs (n=98) were recruited from the Oncology Department in Nanfang Hospital. The Pittsburgh Sleep Quality Index (PSQI), consisting of seven components that evaluate subjective sleep quality, sleep latency, duration of sleep, sleep efficiency, sleep disturbances, sleep medication usage and daytime dysfunction, was used to assess sleep quality. Using the tool of Zung Self-rating Anxiety Scale (SAS) and Zung Self-rating Depression Scale (SDS), we tested the patients' status of anxiety and depression, respectively. RESULTS: The prevalences of poor sleep quality, anxiety and depression in patients were 56.1%, 48.9% and 56.1%, respectively, while those in FCs were 16.3%, 32.6% and 25.5%, respectively. Patients had higher PSQI, SAS and SDS scores than did FCs (p<0.05). Significant correlations were found between the patients' and FCs' scores of PSQI/SAS/SDS (p<0.05). Multivariate Cox regression analyses indicated that sleep disturbances in patients (HR 0.413, 95% CI 0.21 to 0.80, p=0.01) and the global PSQI score of FCs (HR 0.31, 95% CI 0.14 to 0.71, p=0.00) were independent risk factors for patients' first-line progression-free survival (PFS). Moreover, patients' sleep latency (HR 2.329, 95% CI 1.36 to 3.96, p=0.00) and epidermal growth factor receptor mutations (HR 1.953, 95% CI 1.12 to 3.38, p=0.01) were significant prognostic factors for their overall survival (OS). CONCLUSIONS: We demonstrated that presence of sleep disturbances in patients with advanced LC and the global PSQI Score of their FCs may be risk predictors for patients' poor first-line PFS. Patients' sleep latency was a potential risk factor for their OS.

Prognostic Implications of Tripartite Motif Containing 24 Expression Levels in Patients with Solid Tumors: A Systematic Review and Meta-Analysis.

Objective: To systematically investigate the prognostic implications of tripartite motif containing 24 (Trim 24) expression levels in Patients with solid tumors. Materials and Methods: Pubmed, Embase, China National Knowledge Infrastructure, and Wanfang databases were searched through December 2017 to identify studies examining the relationship between Trim 24 expression levels and outcomes in solid tumor patients. The hazard ratios (HRs) with corresponding 95% confidence intervals were used to evaluate the association between Trim 24 and overall survival (OS). Results: Ten studies with 1370 patients were included. The overall pooled prevalence for Trim 24 overexpression was 59.0% (p < 0.01). Moreover, the pooled HR of Trim 24 for OS was 0.43 (p = 0.04) by univariate analysis in 10 articles (1370) and 0.62 (p = 0.08) by multivariate analysis in 5 studies (845). Trim 24 over-expression was associated with tumor invasiveness (odds ratio [OR] = 2.05, p < 0.01) and tumor-node-metastasis stage (OR = 2.42, p = 0.03). Conclusions: This study demonstrated that Trim 24 expression levels may be a useful prognostic biomarker in patients with solid tumors.

AAV2/IL-12 gene delivery into dendritic cells (DC) enhances CTL stimulation above other IL-12 applications: Evidence for IL-12 intracrine activity in DC.

Adoptive transfer of antigen-specific cytotoxic T lymphocytes (CTL) holds significant promise in treating cancer and Th1 response cytokines are critical for their stimulation. Recently we reported that interleukin 7-(IL-7) and interferongamma-(IFNγ) autocrine/T cell gene delivery resulted in superior ex vivo CTL stimulation over paracrine/DC delivery. IL-12 is yet another important Th1 cytokine which affects both DC and T cells. Here, using adeno-associated virus Type 2 (AAV2) gene delivery, IL-12-paracrine/DC gene delivery gave significantly superior stimulation of carcinoembryonic antigen (CEA)-specific CTL killing over that induced by autocrine gene delivery (or exogenous IL-12 addition). This is surprising as both AAV2/IL-12-treated T cells and DC secreted approximately the same level of IL-12. Paracrine IL-12 gene delivery also resulted in highest IL-12/IL-10 secretion ratio by DC and highest CD40, CD80, CD83 and CD86 expression. Moreover, AAV2/IL-12-DC stimulated the highest T-cell IFNγ production, highest T cell proliferation, highest CD69+/CD8+ levels, and lowest level of CD25+/CD4+ Treg. These data strongly suggest that the primary activity of IL-12 during CTL generation is upon the DC. These data are also consistent with there being novel activity for IL-12 within the DC itself, not involving its surface receptor; an "intracrine" activity. Given the plethora of IL-12 studies, these data also suggest that this gene delivery comparison approach could be useful for uncovering new cytokine activities and mechanism(s) of action gone unrecognized by conventional immunologic assays. Finally, these data further suggest AAV2/IL-12 intracrine gene delivery into DC may have utility in immunotherapy protocols involving antigen-specific CTL.

[Ultrasound-mediated microbubble destruction enhances LMP-1 gene transfection into dendritic cells in vivo].

OBJECTIVE: To investigate the transfection efficiency and the optimal conditions of delivering latent membrane protein-1 (LMP-1) gene to dendritic cells (DCs) by ultrasound exposure combined with contrast agent. METHODS: Human DCs were cultured in vivo and transfected with the recombinant plasmid pEGFP-C3-LMP1 under varying conditions including ultrasound intensities, exposure time and microbubble contrast agent concentration. The transfection efficiency was assessed by fluorescent microscopy and flow cytometry, and the cell viability by trypan blue exclusion test. RESULTS: An exposure time of 60 s at MI 1.0 with a microbubble contrast agent concentration of 20% resulted in the optimal effect of delivering the recombinant plasmid pEGFP-C3-LMP1 into the DCs, with a transfection efficiency of (14.37∓2.12)%. Over 90% of the transfected cells were viable after the transfection. CONCLUSION: Microbubble contrast agent combined with ultrasound exposure can enhance the delivery of recombinant plasmid pEGFP-C3-LMP1 into the DCs.

Autocrine, not paracrine, interferon-gamma gene delivery enhances ex vivo antigen-specific cytotoxic T lymphocyte stimulation and killing.

The adoptive transfer of antigen-specific cytotoxic T lymphocytes (CTL) shows promise in the treatment of cancer and infectious diseases. We utilize adeno-associated virus-(AAV-) based antigen gene-loaded dendritic cells (DCs) to stimulate such antigen-specific CTL. Yet further improvements in CTL stimulation and killing may result by gene delivery of various Th1-response interferons/cytokines, such as interferon gamma (IFN-gamma), as the delivered gene can continuously produce that interferon. However which immune cell type should optimally express IFN-gamma is unclear as the phenotypes of both DC and T cells are enhanced by it. Here, we used AAV to compare and contrast IFN-gamma gene delivery into DC or T cells, and versus the addition of exogenous IFN-gamma, for stimulating carcinoembryonic antigen-(CEA-) specific CTL. It was found that AAV/IFN-gamma delivery into T cells (autocrine) resulted in T cell populations with the highest CD8(+)/CD4(+) ratio, highest IFN-gamma(+)/IL-4(+) ratio, highest CD69(+),CD8(+) levels, and lowest CD4(+)/CD25(+) levels, all consistent with the strongest Th1 response. Most importantly, AAV/IFN-gamma transduction of T cells resulted in antigen-specific T cell populations with the highest killing capabilities, 49% above other treatments. These data strongly suggest that AAV/IFN-gamma autocrine gene delivery into T cells is worthy of further study towards maximizing the generation of antigen-specific anticancer CTL killers.

TRPM7 regulates the migration of human nasopharyngeal carcinoma cell by mediating Ca(2+) influx.

Ion channels are involved in various physiologic and pathologic processes, including the migration of tumor cells that is required for metastasis. To determine whether transient receptor potential melastatin 7 (TRPM7) Ca(2+) channels play an important role in the migration of tumor cells, we examined the potential function of TRPM7 channels in the migration of 5-8F and 6-10B human nasopharyngeal carcinoma cells. The migratory potential of 5-8F cells was significantly decreased by extracellular Ca(2+) chelator (EGTA), TRPM7 inhibitors (La(3+), 2-APB), or TRPM7 knockdown. Conversely, the addition of TRPM7 activator Bradykinin and overexpression of TRPM7 promoted the migration of 5-8F and 6-10B cells. Furthermore, the sustained Ca(2+) influx regulated by TRPM7 activated release of Ca(2+) stores via ryanodine receptors by a calcium-induced calcium release (CICR) mechanism. This study suggests, first, that Ca(2+) influx is required for the migration of human nasopharyngeal carcinoma 5-8F cells. Second, and more importantly, it identifies TRPM7 as a novel potential-regulator of the Ca(2+) influx that allows migration of 5-8F cells. TRPM7, therefore, might have potential as a prognostic indicator and as a therapeutic target in nasopharyngeal carcinoma.

Comparison of AAV/IL-7 autocrine (T cell) versus paracrine (DC) gene delivery for enhancing CTL stimulation and function.

Adoptive transfer of antigen-specific cytotoxic T lymphocyte (CTL) into patients holds promise in treating cancer. Such anti-cancer CTL are stimulated by professional antigen-presenting dendritic cells (DC). We hypothesize the gene delivery of various Th1-response cytokines, such as interleukin 7 (IL-7), should further enhance CTL stimulation and activity. However, the issue as to which cell type, DC (paracrine) or the T cell (autocrine), should express a particular Th1 cytokine gene for optimal CTL stimulation has never been addressed. We used adeno-associated virus-2 (AAV) to compare delivery of IL-7 and IL-2 genes into DC or T cells and to exogenous commercial cytokines for generating robust carcinoembryonic antigen (CEA)-specific CTL. AAV/IL-7 transduction of T cells (autocrine delivery) generated CTL with the highest killing capability. Consistent with this, AAV/IL-7 delivery generated T cell populations with the highest proliferation, highest interferon gamma expression, highest CD8(+):CD4(+) ratio, highest CD8(+), CD69(+) levels, and lowest CD4(+), CD25(+) (Treg) levels. These data are consistent with higher killing by the AAV/IL-7-altered CTL. These data strongly suggest that IL-7 autocrine gene delivery is optimal for CTL generation. These data also suggest Th1 cytokine autocrine versus paracrine delivery is an important issue for immuno-gene therapy and uncovers new questions into cytokine mechanism of action.

[Clinical value of (18)F-FDG positron emission tomography-computed tomography in local liver neoplasm ablation].

OBJECTIVE: To assess the value of (18)F-fluoro-2-deoxy-D-glucose positron emission tomography-computed tomography ((18)F-FDG PET-CT) in ultrasound-guided local ablation of malignant liver tumors. METHODS: Thirteen patients with 35 local residual tumor foci following previous tumor ablation underwent (18)F-FDG PET-CT and ultrasound-guided local ablation with intratumoral alcohol injection. RESULTS: After the second local ablation guided by (18)F-FDG PET-CT and ultrasound, radioactive defects were detected in the corresponding location in 31 of the 35 residual foci, and after the third local ablation, the other 4 foci also showed radioactive defects. CONCLUSION: (18)F-FDG PET-CT can sensitively and accurately identify tissue necrosis and residual tumors, and serves as an excellent approach for ultrasound-guided local ablation of local residual tumors.

[rAAV/BA46-transfected dendritic cells can induce specific cellular immunity].

OBJECTIVE: To study the feasibility of transfecting breast cancer BA46 gene into dendritic cells (DCs) using adeno-associated virus (AAV) to induce specific cellular immunity. METHODS: Mononuclear cells (DC precursor) were isolated from the peripheral blood of healthy donors by density gradient centrifugation and infected with rAAV/BA46/Neo virus stock (transfection group) or pulsed with 293 cell lysate (control group). In both groups, maturation of the DC precursor was induced by granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukin-4 (IL-4) and tumor necrosis factor-alpha(TNF-alpha). On day 7, the DCs were collected and mixed with T cells at the ratio of 1 to 20 to induce cytotoxic T lymphocytes (CTL). The capacity of the DCs in stimulating T lymphocyte proliferation was assessed using (3)H-thymidine incorporation assay. The expressions of interferon-gamma (IFN-gamma), IL-4, CD4, CD8, CD25 and CD69 in the CTLs were analyzed with cytometry, and the cytotoxicity of the CTLs was evaluated with (51)Cr-release assay using BA46-positive breast cancer cell line Hs578T as the target. RESULTS: The DCs transfected with BA46 gene exhibited potent capacity to stimulate T lymphocyte proliferation. The CTL population induced by the transfected DCs expressed high levels of CD8, CD69 and IFN-gamma, and showed strong cytotoxicity against BA46-positive breast cancer cell line Hs578T, which was BA46 antigen-specific and MHC-limited. CONCLUSION: The success in BA46 gene transfer in the DCs that induce specific cellular immunity provides the experimental basis for breast cancer immunotherapy using genetically modified cells.

[Construction of adenoviral vector encoding soluble human TNFRI-IgGFc cDNA and its expression in human bronchial epithelial cells].

OBJECTIVE: To construct a recombinant adenovirus vector carrying soluble extracellular region of tumor necrosis factor alpha receptor I-IgGFc (sTNFRI-IgGFc) and express the fusion protein in human bronchial epithelial HBE135-E6E7 cells. METHODS: sTNFRI-IgGFc fusion gene was subcloned into the adenovirus shuttle plasmid pDC316, which was co-transfected with helper plasmid pBHGloxPE1,3Cre into HEK293 cells. The recombinant adenovirus (Ad-sTNFRI-IgGFc) was generated by homologous recombination of the 2 plasmids in HEK293 cells. After identification with PCR, Ad-sTNFRI-IgGFc was amplified and purified, and its titer measured using TCID50 assay. The transcription and expression of sTNFRI-IgGFc gene in the transfected HBE135-E6E7 were detected by RT-PCR and immunohistochemistry. RESULTS: Ad-sTNFRI-IgGFc was successfully constructed with a viral titer of 3 x 10(10) TCID50/ml. The expression of sTNFRI-IgGFc mRNA and protein was confirmed in the transfected HBE135-E6E7 cells. CONCLUSION: The constructed Ad-sTNFRI-IgGFc can effectively infect HBE135-E6E7 cells for efficient expression of sTNFRI-IgGFc protein, which antagonizes the cytolytic effect of TNFalpha in L929 cells, suggesting the potential of adenovirus expressing sTNFRI-IgGFc for local treatment of asthma.

[Effects of two different antigen-loading methods on the activity of dendritic cell vaccine for colorectal carcinoma cell inhibition in vitro].

OBJECTIVE: To observe the changes in the activity of dendritic cells (DCs) after carcino-embryonic antigen (CEA) gene transfection mediated by recombinant adeno-associated virus type2 (rAAV) and tumor cell lysate. METHODS: Immature DCs isolated from peripheral blood monocytes of HLA-A11-positive healthy volunteers were infected with the rAAV carrying CEA gene or loaded with tumor cell lysate. The surface markers of the DCs such as CD40, CD 1alpha, and CD86 were analyzed by flow cytometry. Interleukin-12 (IL-12) in the supernatants of DCs and interferon-gamma (IFN-gamma) released by the cytotoxic T lymphocytes (CTLs) were determined by ELISA detection kit. The specific killing activity of CTL against LoVo cells was assessed by MTT assay. RESULTS: The DCs following antigen loading with the two methods both highly expressed CD40, CD86 and IL-12, and induced specific CTL that specifically recognized and killed LoVo cells, but the killing effect resulting from rAAV infection of the DCs was much better than that induced by tumor cell lysate loading. CONCLUSION: Both methods of antigen loading can induce mature DCs from peripheral blood monocyte cells, but rAAV infection of the DCs can be more effective than tumor cells lysate loading. DCs infected with rAAV may have the potential to serve as an adjuvant immunotherapy for patients with colorectal carcinoma.

[Cytotoxicity of cytotoxic T lymphocytes induced by keratin 19-sensitized dendritic cells against MCF-7 cells in vitro].

OBJECTIVE: To investigate the cytotoxicity of cytotoxic T lymphcytes (CTL) induced by keratin 19 (K19)-sensitized dendritic cells (DC) against MCF-7 cells in vitro. METHODS: DC isolated from peripheral blood mononuclear cells were cultured in vitro and sensitized by K19 peptide to generate specific CTL. The phenotypes and intracytoplasm cytokine of DC were analyzed by flow cytometry. Mixed leukocyte responses were evaluated by (3)H-TdR incorporation assay. The T cells were generated by DC pulsed with K19 antigen and T cell cytotoxicity was measured by (51)Cr release assay. RESULTS: The DCs derived from peripheral blood mononuclear cells expressed high levels of CD40, CD86, CD80 and CD83. Mixed leukocyte responses induced by the DCs pulsed with K19 was stronger than that induced by naive DCs (P<0.01). The cytotoxicity rate of CTL induced by the sensitized DCs was higher than that of CTL induced by naive DCs (P<0.01). The cytotoxity activity was enhanced by increasing the effector/target ratio (P<0.01). CONCLUSION: After sensitization with K19 antigen, the DCs can stimulate T cell proliferation and induce cytotoxicity activity against MCF-7 cells. Increased effector/target ratio may enhance the cytotoxity activity. Sensitized DCs possess the potential for amplification in immunotherapy of carcinoma.

[Immunostimulatory effect of human peripheral blood dendritic cells transfected by adeno-associated virus containing Her2/neu gene].

OBJECTIVE: To observe the immunostimulatory effect of human peripheral blood dendritic cells (DCs) transfected by Her2/neu gene delivered by adeno-associated virus. METHODS: The mononuclear cells in healthy donors were isolated by Ficol-Hypaque density gradient separation and divided into transfection group and control group without transfection by the recombinant virus. The cells were initially cultured in RPMI 1640 medium supplemented with 10% AB human serum, followed by addition of granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukin (IL)-4 and tumor necrosis factor (TNF)-alpha into the medium. The surface markers of DC were detected by flow cytometry, allogeneic mixed lymphocyte reaction assayed by 3H-TdR incorporation and the specific killing activity of T cells evaluated by MTT assay. RESULTS: The expression rates of CD1a, CD86 and CD83 of the transfected DCs were 98.10%, 99.42%, and 84.59%, and those of the non-transfected DCs were 92.69%, 98.07%, and 82.72%, respectively, showing no obvious differences between the two groups. The rates of CD4 and CD80 expression were 61.02% and 97.61%, respectively, in the transfected DCs, significantly higher than those in the non-transfected cells (36.19% and 55.5%). Both groups of DCs stimulated a strong T cell proferation response. The transfected DCs were capable of inducing specific killing of the target tumor cells, with the highest killing rate of (39.7+/-7.2)%. CONCLUSION: The immunostimulatory effect of human peripheral blood DCs can be enhanced by Her2/neu gene transfection mediated by adeno-associated virus.

[Clinical observation of the efficacy of navelbine combined with ifosfamide and cisplatin in the treatment of advanced breast cancer].

OBJECTIVE: To observe the efficacy of navelbine combined with ifosfamide and cisplatin in the treatment of advanced breast cancer. METHODS: Forty cases of pathologically confirmed stages III-IV breast cancer were treated with combined chemotherapy including intravenously infused navelbine (25 mg/m2, days 1 and 8), ifosfamide (1.3 g/m2, days 1 to 5) and cisplatin (20 mg/m2, days 1-5), administered in cycles of 21 days. The therapeutic effect was assessed after at least two cycles were completed. RESULTS: Complete remission was achieved in 6 cases and partial remission in 16, with an overall responding rate of 55%. The main toxicity was bone marrow suppression manifested by grades III-IV leukopenia occurring at a rate of 40%. CONCLUSION: Navelbine in combination with ifosfamide and cisplatin is effective and safe in the treatment of advanced breast cancer.

[Clinical study of serum tissue polypeptide-specific antigen in breast cancer].

OBJECTIVE: To investigate the expression of serum tissue polypeptide-specific antigen (TPS) in breast cancer patients and its clinical value in such cases. METHODS: Altogether 160 subjects (90 patients with breast cancer, 40 with benign breast lesions, and 30 healthy subjects) were enrolled in this study. The serum TPS and CA153 levels were measured by ELISA in all the subjects. RESULTS: The levels and positivity rate of serum TPS and CA153 in breast cancer group were significantly higher than those in the normal subjects group and benign lesion group (P<0.01), and became even higher as the malignancy progressed. High serum TPS level was detected in the cancer patients in stage I. Serum TPS level was the most sensitive to bone metastasis of the malignancy, but its highest levels occurred in cases of lymphoid node metastasis (P<0.05). In patients who responded favorably to the treatment, serum TPS and CA153 levels were significantly reduced (P<0.05), but the reduction in TPS levels tended to be more obvious (P<0.05). CONCLUSION: Serum TPS can be used as a very useful and sensitive tumor marker in the diagnosis of breast cancer, especially in case of bone metastasis, and may be of great value in clinical decision-making and assessment of therapeutic effect.

Emergency management of respiratory failure by endotracheal intubation through the nose under the guidance of portable bedside fibrobronchoscope.

OBJECTIVE: To investigate the value of portable fibrobronchoscope in the management of respiratory failure by endotracheal intubation through the nose. METHODS: Fifty-eight patients with acute or chronic respiratory failure received mechanical ventilation by endotracheal intubation through the nose under the guidance of portable fibrobronchoscope. RESULTS: Intubation was successfully performed in all the patients in a single attempt (which took 30 min to 5 min) without interruption of autonomous breathing or incurring laryngospasm or cardiac arrest. After mechanical ventilation for 30 min, conspicuous improvement of respiratory failure was observed in all the cases. CONCLUSION: With convenient and easy manipulation, portable fibrobronchoscope provides quick and accurate guidance for endotracheal intubation through the nose in the emergency management of respiratory failure.

Preparation and identification of monoclonal antibodies against human brain-derived neurotrophic factor.

OBJECTIVE: To prepare monoclonal antibodies (mAbs) against human brain-derived neurotrophic factor (BDNF). METHODS: BALB/C mice were immunized with purified BDNF protein in conjunction with acid-treated Salmonella (antigen:thallus=1:5), and mAbs were subsequently derived by hybridoma technique. RESULTS: Three hybridoma cell lines secreting anti-BDNF mAbs were obtained, designated as B1, B2 and 4D1 respectively and all categorized into IgG1 subtype. The titers of the mAbs in the ascitic fluid of the rats ranged from 1x10(6) to 1x10(5), and their relative affinities were B2>B1>4D1. CONCLUSION: mAbs against BDNF are successfully prepared, which can be instrumental for further study of the expression and distribution of BDNF in vivo, and the detection of BDNF produced by genetic engineering.