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Background: Huangqi Guizhi Wuwu decoction (HQGZWW) exhibits good effects when administered to treat multiple sclerosis (MS) and its animal model, experimental autoimmune encephalomyelitis (EAE). Understanding the precise mechanism of this decoction is thus important. Based on the findings of our previous study, the aim of the present study was to understand the role of antigen-specific CD8+ T-cells on the pathogenesis of MS/EAE when HQGZWW is administered as treatment. Methods: Myelin oligodendrocyte glycoprotein (MOG) 35-55-induced mice were administered distilled water, prednisone, and high dose or low dose HQGZWW. After purified CD4+ and CD8+ T-cells were stimulated with the MOG35-55 peptide, proliferation and cytokine secretion assays were performed. To establish the adoptive transfer EAE model, naive mice were injected with MOG35-55 -CD8+ or CD4+ T-cells. Results: Significant improvements in EAE score and pathology were observed in the high dose HQGZWW and prednisone groups. Compared to the low dose HQGZWW and distilled water groups, lower antigen-specific responses, lower levels of interferon-gamma, and higher levels of interleukin (IL)-4 and IL-10 from CD8+ and CD4+ T cells were observed in the high dose HQGZWW and prednisone groups. Finally, the EAE score was observed to be similar between the high dose HQGZWW group and prednisone group; however, this finding was not observed in the low dose HQGZWW group. Conclusion: Our findings suggest that high dose HQGZWW has similar effects on cell proliferation, cytokine secretion, and EAE score to prednisone, while low dose HQGZWW does not have such effect. The protective role of HQGZWW against EAE might thus depend on the Th2 cytokine secretion profile induced by either MOG35-55 specific CD8+ or CD4+ T-cells.

Background: The pathogenesis of multiple sclerosis (MS) is mediated primarily by T cells, but most studies of MS and its animal model, experimental autoimmune encephalomyelitis (EAE), have focused on CD4+ T cells. The aims of the current study were to determine the pathological interrelationship between CD4 and CD8 autoreactive T cells in MS/EAE. Methods: Female C57BL/6 mice (n =20) were induced by myelin oligodendrocyte glycoprotein (MOG) 35-55 peptide. At 14 days after immunization, T cells were isolated from the spleen and purified as CD4+ and CD8 + T cells by using CD4 and CD8 isolation kits, and then the purity was determined by flow cytometric analysis. These cells were stimulated by MOG35–55 peptide and applied to proliferation assays. The interferon-gamma (IFN-γ) and interleukin (IL)-4 secretion of supernatant of cultured CD4 + and CD8 + T cells were measured by enzyme-linked immunosorbent assays (ELISA). For adoptive transfer, recipient mice were injected with MOG35–55-specific CD8+ or CD4+ T cells. EAE clinical course was measured by EAE score at 0–5 scale and spinal cord was examined by staining with hematoxylin and eosin and Luxol fast blue staining. Results: CD8+CD3+ and CD4+CD3 + cells were 86% and 94% pure of total CD3+ cells after CD8/CD4 bead enrichment, respectively. These cells were stimulated by MOG35–55 peptide and applied to proliferation assays. Although the CD8+ T cells had a generally lower response to MOG35–55 than CD4+ T cells, the response of CD8+ T cells was not always dependent on CD4. CD8+ T cell secreted less IFN-γ and IL-4 compared with CD4+ T cells. EAE was induced in wildtype B6 naive mice by adoptive transfer of MOG35–55-specific T cells from B6 active-induced EAE (aEAE) mice. A similar EAE score and slight inflammation and demyelination were found in naive B6 mice after transferring of CD8 + T cells from immunized B6 mice compared with transfer of CD4 + T cells. Conclusion: Our data suggest that CD8+ autoreactive T cells in EAE have a lower encephalitogenic function but are unique and independent on pathogenic of EAE rather than their CD4+ counterparts. Key words: Autoimmune; CD8-Positive T-Lymphocytes; Encephalomyelitis; Experimental; Multiple Sclerosis; Myelin-Oligodendrocyte Glycoprotein

Cryptococcus is an opportunistic pathogenic fungus and is the major cause of fungal meningitis. The cryptococcal antigen (CrAg) lateral flow assay (LFA) is an immunochromatographic test system that has simplified diagnosis as a point-of-care test. In this study, we evaluated the diagnostic performance of Cryptococcal capsular polysaccharide detection FungiXpert (Genobio Pharmaceutical, Tianjin, China) using serum and cerebrospinal fluid (CSF) samples for the diagnosis of cryptococcosis and investigated the cross-reaction of the assays to pathogenic fungi and bacterium by comparing it to the U.S. Food and Drug Administration (US FDA)-approved IMMY CrAg LFA. Eighty CSF and 119 serum/plasma samples from 158 patients were retrospectively collected to test for qualitative or semi-quantitative detection of CrAg. Cross-reaction of the assays was tested using 28 fungi and 1 bacterium. Compared to IMMY CrAg LFA, the FungiXpert LFA demonstrated 99.1% sensitivity and 98.9% specificity in the qualitative test. In the 96 semi-quantitative CrAg assay results, 39 (40.6%) test titers of FungiXpert LFA were 1–2 dilutions higher than those of IMMY CrAg LFA. The Intraclass Correlation Coefficient of the Semi-quantitative results of CrAg titer tests via the two assays was 0.976. Similar to IMMY CrAg LFA, FungiXpert LFA showed cross-reactivity with Trichosporon asahii. Compared with the IMMY CrAg LFA, the FungiXpert LFA showed an equal, yet, excellent performance. However, it is important to note that these two assays have potential cross-reactivity to T. asahii when diagnosing patients. FungiXpert LFA is a rapid screening method for the effective and practical diagnosis and treatment of cryptococcosis. Lay summary The FungiXpert LFA was developed to diagnose fungal meningitis caused by Cryptococcus yeasts, by using serum or cerebrospinal fluid. It was compared to an existing lateral flow assay (LFA). The FungiXpert LFA performed well in qualitative and semi-quantitative tests.

This paper presents a compact dual-band ±45° dual-polarized base-station array based on embedded filtering elements. The proposed antenna array consists of 9 dual-band antenna units, which operate at the lower band (LB) of 790-862 MHz and the higher band (HB) of 880-960 MHz with a small frequency spacing. In each unit, one antenna element is embedded into the other one, featuring a very compact size. To alleviate the serious cross-band mutual coupling between the two elements, the LB patch antenna generates a boresight radiation null at its higher band edge, and the HB element is designed with a band-stop response at its lower band edge. In this way, high cross-band isolation between the two embedded elements can be obtained. For demonstration, a compact dual-band ±45° dual-polarized filtering antenna array with 9 units is designed and fabricated to fit the specification of LB and HB bands (|S11

A titanium alloy containing continuous oxygen gradient was prepared by powder metallurgy (P/M) and the composition–property relationship was studied on a single sample. The alloy was sintered with layered powder of different oxygen contents via vacuum sintering and spark plasma sintering (SPS), respectively. After subsequent heat treatments, high-throughput characterizations of the microstructures and mechanical properties by localized measurements were conducted. The Ti-7% Mo (molar fraction) alloy with an oxygen content ranging from 1.3×10−3 to 6.2×10−5 (mass fraction) was obtained, and the effects of oxygen on the microstructural evolution and mechanical properties were studied. The results show that SPS is an effective way for fabricating fully dense Ti alloy with a compositional gradient. The average width of α′ phase coarsens with the increase of the content of oxygen. The content of α″ martensitic phase also increases with the content of oxygen. At oxygen contents of 3×10−3 and 4×10−3 (mass fraction), the Ti alloys present the lowest microhardness and the lowest elastic modulus, respectively. The results also indicate that the martensitic phases actually decrease the hardness of Ti-7Mo alloy, and oxygen effectively hardens the alloy by solid solution strengthening. Therefore, the high-throughput characterization on a microstructure with a gradient content of oxygen is an effective method for rapidly evaluating the composition–property relationship of titanium alloys.

Titanium-based metal-metal composites show promises in many applications. In this work, iron-containing titanium-based composites were fabricated by using powder metallurgy method and rotary swaging process. Titanium-based composites with steel addition were sintered at temperature below Ti Fe eutectic temperature. Increase of the steel addition to 23% resulted in formation of Ti Fe intermetallic and density decrease in the composites. Further, the as-sintered composites present relatively good workability so hot rotary swaging of the as-sintered composites can be conducted. Steel particles were diffused into the Ti matrix and the composite transformed to a discontinuous β-Ti fiber-reinforced structure. As a result, mechanical properties (in axial direction) of the swaged composites were significantly improved. The tensile strength and elongation reached 1360 MPa and 9.3%, respectively.

AIM: To systematically review the evidence for the effectiveness of fast-track program vs traditional care in laparoscopic or open surgery for gastric cancer. METHODS: PubMed, Embase and the Cochrane library databases were electronically searched for published studies between January 1995 and April 2013, and only randomized trials were included. The references of relevant studies were manually searched for further studies that may have been missed. Search terms included “gastric cancer”, “fast track” and “enhanced recovery”. Five outcome variables were considered most suitable for analysis: postoperative hospital stay, medical cost, duration to first flatus, C-reactive protein (CRP) level and complications. Postoperative hospital stay was calculated from the date of operation to the date of discharge. Fixed effects model was used for meta-analysis. RESULTS: Compared with traditional care, fast-track program could significantly decrease the postoperative hospital stay [weighted mean difference (WMD) = -1.19, 95%CI: -1.79--0.60, P = 0.0001, fixed model], duration to first flatus (WMD = -6.82, 95%CI: -11.51--2.13, P = 0.004), medical costs (WMD = -2590, 95%CI: -4054--1126, P = 0.001), and the level of CRP (WMD = -17.78, 95%CI: -32.22--3.35, P = 0.0001) in laparoscopic surgery for gastric cancer. In open surgery for gastric cancer, fast-track program could also significantly decrease the postoperative hospital stay (WMD = -1.99, 95%CI: -2.09--1.89, P = 0.0001), duration to first flatus (WMD = -12.0, 95%CI: -18.89--5.11, P = 0.001), medical cost (WMD = -3674, 95%CI: -5025--2323, P = 0.0001), and the level of CRP (WMD = -27.34, 95%CI: -35.42--19.26, P = 0.0001). Furthermore, fast-track program did not significantly increase the incidence of complication (RR = 1.39, 95%CI: 0.77-2.51, P = 0.27, for laparoscopic surgery; and RR = 1.52, 95%CI: 0.90-2.56, P = 0.12, for open surgery). CONCLUSION: Our overall results suggested that compared with traditional care, fast-track program could result in shorter postoperative hospital stay, less medical costs, and lower level of CRP, with no more complications occurring in both laparoscopic and open surgery for gastric cancer.

To analyze the etiology of infections in the wounded victims of Wenchuan Earthquake.2135 smears of secretion were made from 1823 hospitalized wounded victims of Wenchuan Earthquake to detect the pathogens. Specimens were delivered to be cultured. The bacteria thus obtained were identified. Drug sensitivity test was conducted.2002 specimens, 1243 specimens of secretion (62.1%), 600 blood specimens (30.0%), 102 specimens of pus or secretion of respiratory tract (5.1%), 45 specimens from catheter (2.2%), and 12 urine specimens (0.5%). Pathogens were found in 725 cases. The top five pathogenic bacteria isolated within 1 month after the quake were Acinetobacter baumannii (16.7%), Staphylococcus haemolyticus (16.7%), Escherichia coli (12.5%), Klebsiella pneumoniae (12.5%), and Candida tropicalis (8.3%), quite different from the pathogen spectrum of the common in-patients within one month before the quake: Escherichia coli (18.2%), Staphylococcus aureus (11.4%), Candida glabrata (11.4%), Klebsiella pneumoniae (11.4%) and Staphylococcus epidermidis (9.1%). The isolation rate of methicillin resistant Staphylococcus aureus after the earthquake was significantly lower, and the isolation rates of extended spectrum beta-lactamase-producing Escherichia coli, Klebsiella pneumoniae, and Klebsiella oxytoca were all significantly higher than those from the common surgical patients before the quake (all P0.05). There were not significant differences in the isolation rates of multi-drug resistant Pseudomonas alcaligenes and Acinetobacter baumannii before and after the quake.Infection is frequent after natural disasters. It is necessary to summarize the changes of spectrum of pathogens and drug-resistant spectrum.

Objective To investigate the analgesia induced by receptin (REC), a chemically modified cobratoxin (CTX, a long-chain postsynaptic alpha -neurotoxin from Thailand cobra venom), and the effects of atropine and naloxone on antinociceptive activity of REC in rodent pain models. Methods REC was administered intraperitoneally (5 mg/kg, 7.07 mg/kg, or 10 mg/kg, i.p.) or intra-cerebral venticularly (62.5 mu g/kg, i.c.v.). The antinociceptive action was determined using the hot-plate test, the acetic acid writhing test and tail flick assay in mice and rats. The involvement of cholinergic and the opioid peptidergic systems in REC-induced analgesia were examined by pretreatment of animals with atropine (Atr; 0.5 mg/kg, i.m. or 10 mg/kg, i.p.) or naloxone (Nal; 3 mg/kg, i.p.). The effect of REC on motor activity was tested using the Animex test in mice. Results REC (5 mg/kg, 7.07 mg/kg or 10 mg/kg, i.p.) exhibited a dose-dependent analgesic action in mice as determined with hot-plate test and acetic acid writhing test. The significant analgesia of REC was seen 2 h to 3 h after its administration. In the rat-tail flick assay, the administration of REC at 62.5 mu g/kg (1/160 of systemic dose; i.c.v.) produced marked analgesic effects. Atropine at 0.5 mg/kg (i.m.), 10 mg/kg (i.p.) or naloxone at 3 mg/kg (i.p.) failed to block the analgesic effects of REC. REC at the highest effective dose of 10 mg/kg did not change the spontaneous mobility of mice. Conclusion These results demonstrate that REC has analgesic effect. This activity appears to be mediated through the peripheral nervous system though central nervous system may contribute to REC' s analgesic effects. The central cholinergic system and opioid peptidergic system appear not to be involved in the antinociceptive action of REC.

Objective To investigate the analgesia induced by cobrotoxin (CT) from venom of Naja naja atra, and the effects of atropine and naloxone on the antinociceptive activity of CT in rodent pain models. Methods CT was administered intraperitoneally (33.3, 50, 75 mu g/kg), intra-cerebral venticularly (2.4 mu g/kg) or microinjected into periaqueductal gray (PAG, 1.2 mu g/kg). The antinoCiceptive action was tested using the hot-plate test and the acetic acid writhing test in mice and rats. The involvement of cholinergic system and the opioid system in CT-induced analgesia was examined by pretreatment of animals with atropine (0.5 mg/kg, im or 10 mg/kg, ip) or naloxone (3 mg/kg, ip). The effect of CT on motor activity was tested using the Animex test. Results CT (33.3, 50 and 75 mu g/kg, ip) exhibited a dosedependent analgesic action in mice as determined with hot-plate test and acetic acid writhing test. In the mouse acetic acid writhing test, the intra-cerebral ventricle administration of CT 2.4 mu g/kg (1/23th of a systemic dose) produced marked analgesic effects. Microinjection of CT 1.2 mu g/kg (1/46th of systemic dose) into the PAG also elicited a robust analgesic action in the hot-plate test in rats. Atropine at 0.5 mg/kg (im) or naloxone at 3 mg/kg (ip) failed to block the analgesic effects of CT, but atropine at 10 mg/kg (ip) did antagonize the analgesia mediated by CT in the mouse acetic acid writhing test. At the highest effective dose of antinociception (75 mu g/kg), CT did not change the spontaneous mobility of mice. Conclusion These results suggest that CT from Naja naja atra venom has analgesic effects. Central nervous system may be involved in CT's analgesic effects and the PAG may be the primary central site where CT exerts its effects. The central cholinergic system but not opioid system appears to be involved in the antinociceptive action of CT.

To study the therapeutic effect of intraperitoneal hyperthermic double-distilled water and cis-diaminodichloro-platinum (DDP) perfusion for cancerous ascites and radical gastrectomy.LACA mice were injected peritoneally with H22 cancer cells (2 × 10(7) tumor cells). Five days later, the mice received treatments with either intraperitoneal perfusion of 37 °C isotonic fluid (group I), or 43 °C simple hyperthermic double-distilled water (group II), isotonic fluid (group III), DDP (group IV) or a combination of the hyperthermic double-distilled water with DDP (group V). A clinical experiment with intraperitoneal hyperthermic double-distilled water perfusion with DDP was carried out from September 1991 through September 1993 with 32 advanced gastric cancer patients who had undergone radical gastrectomy.In comparison with the untreated control group of cancer cell-bearing LACA mice, the mice in all treatment groups showed near complete obliteration of cancer cells in the peritoneal cavity, markedly reduced ascites, prolonged survival times, and reduced growth of peritoneal cancerous nodes. In the clinical experiment, all 32 patients with advanced carcinoma had achieved satisfactory results at the 1-year follow-up, but had unsatisfactory results at the 2-year follow-up.The intraperitoneal hyperthermic double-distilled water perfusion with DDP inhibited the occurrence of ascites in LACA mice bearing cancer cells, and prolonged the lifetime of patients with gastric cancer who had undergone radical gastrectomy.

AIM: To seek the X associated protein (XAP) with the constructed bait vector pAS2-1X from normal human liver cDNA library. METHODS: The X region of the HBV gene was amplied by PCR and cloned into the eukaryotic expression vector pAS2-1.The reconstituted plasmid pAS2-1X was transformed into the yeast cells and the expression of X protein (pX) was confirmed by Western blot analysis. Yeast cells were cotransformed with pAS2-1X and the normal human liver cDNA library and were grown in selective SC/-trp-leu-his-ade medium, the second screen was performed with the LacZ report gene. Furthermore, segregation analysis and mating experiment were performed to eliminate the false positive and the true positive clones were selected for PCR and sequencing. RESULTS: Reconstituted plasmid pAS2-1X including the anticipated fragment of X gene was proved by auto-sequencing assay. Western blot analysis showed that reconstituted plasmid pAS2-1X expressed BD:X fusion protein in yeast cells. Of 5 × 106 transformed colonies screened, 65 grew in the selective SC/-trp-leu-his-ade medium, 5 scored positive for β-gal activity, and only 2 remaining clones passed through the segregation analysis and mating experiment. Sequence analysis identified that two clones contained similar cDNA fragment: GAACTTGCG. CONCLUSION: The short peptide (glutacid-leucine-alanine)is a possible required site for XAP binding to pX. Normal human liver cDNA library has difficulties in expressing the integrated XAP on yeast cells.