[Establishment and consistency verification of the standard operation procedure for laboratory detection of immunoglobulin G oligoclonal bands in cerebrospinal fluid].

Objective: To establish the standard operation procedure (SOP) for detection of oligoclonal band (OCB) in cerebrospinal fluid (CSF) and verify consistency by using this SOP in different laboratories. Methods: The SOP for detection of CSF-OCB fluid was successfully established by an expert feedback approach. Neuroimmunology laboratories in 3 representative Chinese three-tier research hospitals were selected for this study, and commercially available protein electrophoresis automation systems and detection SOP were set up. The quality control product was provided by the College of American Pathologists (CAP) and Sebiacompany were used for interior quality and compared to each other, respectively. Seventeen serum and CSF paired samples were tested and compared using the same SOP. Kappa test or Kendall W test were adopted to evaluate the inter-laboratory consistency among different hospitals. Results: The results of repeated testings in a single hospital suggested that the 2-and 4-fold dilution for CSF-OCB were reported as positive, while 64-and 128-fold dilution were reported as negative. Positive or negative inconsistencies were reported in 8-, 16-, and 32-fold dilution. After increasing the number of repetitions, the results showed that both 16-and 32-fold dilution were reported as negative, and 8-fold dilution exhibited negative results (2 positive results for 40 repetitions, coincidence rate=95%).The results of multi-center inter-laboratory quality assessment showed that the detection consistency rate among 3 hospitals was 100% (Kappa value =1). Conclusions: The SOP to detect CSF-OCB established in this study demonstrates a good repeatability and stability. Therefore,such SOP would be a good reference for diagnostic laboratories to detect CSF-OCB in China.

[Analysis of magnetic resonance imaging characteristics in Chinese patients with myelin oligodendrocyte glycoprotein antibody associated disorders].

Objective: To investigate the magnetic resonance imaging (MRI) characteristics in the brain and spinal cord of Chinese patients with myelin oligodendrocyte glycoprotein antibodies associated diseases (MOGAD). Methods: Forty nine MOGAD patients with seropositive MOG-IgG and 58 AQP4-IgG positive patients were enrolled in this study. The characteristics of brain and spinal cord MRI were retrospectively analyzed. Results: There was no significant difference in the proportion of abnormal brain MRI of the two groups (69.4% vs 65.5%, P=0.177) , while the proportion of abnormal spinal cord MRI of the AQP4-IgG positive group was significantly higher than that in the MOG-IgG positive group (84.5% vs 36.7%, P=0.001) . The proportion of MOG-IgG positive patients with subcortical white matter lesions and large lesions in the brain MRI was significantly higher than that in AQP4-IgG positive group (48.9% vs 13.8%, P=0.003, 46.9% vs 12.1%, P=0.000) . The longitudinally extensive transverse myelitis in spinal cord MRI of AQP4-IgG positive group was significantly higher than that in the MOG-IgG group (70.7% vs 24.5%, P=0.002) . In addition, the proportion of MOG-IgG positive child patients with large lesions in the brain was significantly higher than that in AQP4-IgG positive child patients (76.9% vs 20.0%, P=0.047) . Conclusion: Demyelinating MRI lesions caused by MOG-IgG are heterogeneous, and could lead to a wide range of clinical phenotypes which is significantly different from those with AQP4-IgG.

A lethal pneumonia model of Acinetobacter baumannii: an investigation in immunocompetent mice.

OBJECTIVES: Acinetobacter baumannii can cause severe nosocomial and community-acquired pneumonia. To study the pathogenesis of A. baumannii and to develop new treatments, appropriate mouse models are needed. Most reported mouse models of pulmonary A. baumannii infection are non-lethal or require mouse immunosuppression to enhance infection. These models are not suitable for studying host immune responses or evaluating immunotherapies. METHODS: The virulence of 30 clinical isolates was assessed in mice. The most virulent isolate, SJZ24, was selected to develop a pneumonia model in immunocompetent mice. The cytokine mRNA expression in the lung was assessed with real-time PCR. The cell infiltration in bronchoalveolar lavage fluid (BALF) after SJZ24 infection was determined by flow cytometry. Vaccine efficacy was assessed using this model. RESULTS: Intratracheal inoculation of SJZ24 (5 × 107 CFU) resulted in death in 100% of the mice (5/5). SJZ24-infected mice showed high bacterial burdens in blood and organs as well as severe lung-tissue damage. Infection with SJZ24 induced increased inflammatory cytokine expression in the lung and increased neutrophil infiltration in BALF. Immunization with inactivated whole cells of SJZ24 showed 100% protection (5/5) against A. baumanni infection in this model. CONCLUSIONS: We established a lethal pneumonia model in immunocompetent mice with hypervirulent A. baumannii isolate SJZ24. This model can be used to study the immune response to A. baumannii infection and to evaluate vaccine efficacy.

Clocked atom delivery to a photonic crystal waveguide.

Experiments and numerical simulations are described that develop quantitative understanding of atomic motion near the surfaces of nanoscopic photonic crystal waveguides (PCWs). Ultracold atoms are delivered from a moving optical lattice into the PCW. Synchronous with the moving lattice, transmission spectra for a guided-mode probe field are recorded as functions of lattice transport time and frequency detuning of the probe beam. By way of measurements such as these, we have been able to validate quantitatively our numerical simulations, which are based upon detailed understanding of atomic trajectories that pass around and through nanoscopic regions of the PCW under the influence of optical and surface forces. The resolution for mapping atomic motion is roughly 50 nm in space and 100 ns in time. By introducing auxiliary guided-mode (GM) fields that provide spatially varying AC Stark shifts, we have, to some degree, begun to control atomic trajectories, such as to enhance the flux into the central vacuum gap of the PCW at predetermined times and with known AC Stark shifts. Applications of these capabilities include enabling high fractional filling of optical trap sites within PCWs, calibration of optical fields within PCWs, and utilization of the time-dependent, optically dense atomic medium for novel nonlinear optical experiments.

Orthotopic Kidney Transplantation in the Rat With the Use of a Sleeve Arterial Anastomosis Method and a Modified Stenting Technique for Renal Veins.

BACKGROUND: We describe a simple and reliable orthotopic kidney transplantation method in rats with the use of sleeve arterial anastomosis and a modified stenting technique for anastomosis of the renal vein (RV). METHODS: Male Fischer and Lewis rats were used as kidney donors and recipients, respectively, and their left kidneys were harvested in situ. In the control rats (n = 30), the renal artery (RA) and RV anastomoses were performed end-to-end with interrupted sutures by means of the conventional technique. In the experimental animals (n = 30), revascularization of the RA was fashioned end-in-end with the use of a modified sleeve anastomosis, the RV was anastomosed end-to-end with the use of a modified stenting technique and interrupted sutures, and the ureter was anastomosed with the use of the end-to-end interrupted suture technique. RESULTS: The arterial anastomosis time in the control group was 8.52 ± 1.1 minutes, and that in the experimental group was 4.7 ± 0.6 minutes (P < .05). The venous anastomosis time in the experimental group was 9.2 ± 1.3 minutes, which also was less than in the control group (11.19 ± 0.78 minutes; P < .05). The warm ischemia time decreased from 26.8 ± 1.3 minutes in the control group to 20.7 ± 0.5 minutes in the experimental group (P < .05). The success rate of 93% at 21 days after grafting was identical in the experimental and control groups. CONCLUSIONS: We developed a modified model of orthotopic kidney transplantation that can significantly reduce the warm ischemia time.

Scleromyxedema secondary to hepatitis C virus and successfully treated with antiviral therapy.

Scleromyxedema (SM) is a chronic and progressive fibromucinous disease with no known etiology. We report a patient with scleromyxedema associated with hepatitis C virus, successfully treated with interferon and ribavirin therapy.

Common genes in coronary artery disease from Europe, Asia and North America regardless of race and lifestyle.

OBJECTIVE: Coronary artery disease (CAD) remains one of the major causes of death worldwide. Despite considerable advances in the prevention and treatment of CAD, its complications, morbidity and mortality still remain very high, and vary widely across different ethnic groups. MATERIALS AND METHODS: To detect genes involved in the development of CAD, we collected gene expression studies in the blood samples of CAD patients from different continents by searching the Gene Expression Omnibus database (GEO), performed a comparative analysis of gene expression between CAD patients and normal controls (NC) in each continent and identified the common set of differentially expressed genes (DEGs) between CAD patients and NC across different continents. PPI networks of the common set of DEGs were established by Cytoscape software to understand their biological role in CAD. RESULTS: A total of 575, 868 and 476 genes were identified to be significantly differentially expressed between CAD patients and NC in Asia, Europe and North America. 24 genes were found common in three different continents, and 6 genes were previously linked to CAD or atherosclerosis. In the PPIs network the significant hub proteins contained IRF4 (Degree = 23), PLAUR (Degree = 17) and HIST1H2AE (Degree = 15). CONCLUSIONS: Not only did we detect gene expression differences in the blood samples between CAD and NC in Asia, Europe and North America population, but analysis of the three population groups revealed a common set of 24 genes regardless of differences related to race, ethnicity, lifestyle, and environmental factor which may provide key factors to understand the pathogenesis of CAD and lead to development of diagnostic markers and/or effective therapeutic strategies.

Mechanism of antitumor activity of a single-chain interleukin-12 IgG3 antibody fusion protein (mscIL-12.her2.IgG3).

We have constructed an antibody interleukin-12 (IL-12) fusion protein (mscIL-12.her2.IgG3) that demonstrates significant antitumor activity against the murine carcinoma CT26-expressing human HER2/neu. We now report that this antitumor activity is dose dependent and comparable to or better than recombinant murine IL-12 (rMuIL-12) using subcutaneous and metastatic models of disease. The antitumor activity of mscIL-12.her2.IgG3 is reduced in Rag2 knockout mice, suggesting that T cells play a role in tumor rejection. In SCID-beige mice, the antitumor activity is further reduced, suggesting that natural killer (NK) cells or macrophages or both are also important. The isotype of the antibody response to HER2/neu is consistent with a switch from a Th2 to a Th1 immune response and the infiltration of mononuclear cell in tumors from mice treated with mscIL-12.her2.IgG3. Immunohistochemistry reveals that mscIL-12.her2.IgG3 is antiangiogenic. Thus, the mechanism of the antitumor activity exhibited by mscIL-12.her2.IgG3 is highly complex and involves a combination of T and NK cell activity, a switch to a Th1 immune response, and antiantiogenic activity. This is the first study comparing the in vivo antitumor activity of an antibody-IL-12 fusion protein and free IL-12. Our results suggest that antibody-IL-12 fusion proteins may be useful for the treatment of human cancer.

A single-chain IL-12 IgG3 antibody fusion protein retains antibody specificity and IL-12 bioactivity and demonstrates antitumor activity.

IL-12 is a heterodimeric cytokine with many actions on innate and cellular immunity that may have antitumor and antimetastatic effects. However, systemic administration of IL-12 can be toxic. Tumor-specific Abs provide a means to selectively target a metastatic/residual nodule and deliver therapeutic quantities of an immunostimulatory molecule like IL-12 with lower systemic levels and ideally, toxicity. We report the construction and characterization of an Ab fusion protein in which single-chain murine IL-12 is fused to an anti-Her2/neu Ab at the amino terminus (mscIL-12.her2.IgG3). The use of single-chain IL-12 in the fusion protein simplifies vector construction, ensures equimolar concentrations of the two IL-12 subunits, and may confer greater stability to the fusion protein. SDS-PAGE analysis shows this 320-kDa protein is secreted and correctly assembled. FACS analysis demonstrates that this fusion protein binds to cells transfected with the Her2/neu Ag, thus retaining Ab specificity; this fusion protein also binds to a cell line and to PHA-activated PBMC that express the IL-12R, thus demonstrating cytokine receptor specificity. T cell proliferation assays and NK cytotoxicity assays demonstrate that this fusion protein exhibits IL-12 bioactivity comparable to recombinant murine IL-12. In vivo studies demonstrate that this fusion protein has antitumor activity. These results are significant and suggest that this IL-12 Ab fusion protein can effectively combine the therapeutic potential of IL-12 with the tumor-targeting ability of the Ab and may provide a viable alternative to systemic administration of IL-12.