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Objective: To purify Mannan-binding lectin (MBL) from human serum and detect its binding ability to several kinds of bacteria common in infectious diseases of children. Methods: MBL was purified from human serum by affinity chromatography on mannan-Sepharose 4B column. Its binding ability to eight species, 97 strains of bacteria was detected by enzyme-linked lectin assay (ELLA). Results: MBL has different binding ability to bacteria and shows strong binding ability to Klebsiella ornithinolytica and Escherichia coli, but shows relatively lower binding ability to Staphylococcus haemolyticus, Enterobacter cloacae and Staphylococcus epidermidis. To different isolates of Klebsiella pneumoniae, Haemophilus influenzae and Staphylococcus aureus, MBL shows quite different binding ability. Conclusions: MBL has different binding ability to different bacteria, and has relatively stronger binding ability to Gram-negative bacteria. Its binding ability to different isolates of certain kinds of bacteria is quite different.

Background: Advanced oxidation protein products (AOPPs), a marker of oxidative stress, are prevalent in many kinds of disorders. Rheumatoid arthritis (RA), mainly resulting from the dysfunction of fibroblast-like synoviocytes (FLSs), is related to oxidative stress. Although the increased levels of AOPPs in RA patients were reported, the effect of AOPPs on FLSs function still remains unclear. Therefore, our study aims to investigate whether AOPPs have an effect on the inflammatory response of FLSs in vitro. Methods: FLSs obtained from both knees of rats were treated with or without AOPPs-modified rat serum albumin (AOPPs-RSA) in vitro. The mRNA and protein expression of tumor necrosis factor (TNF)-a, interleukin(IL)-1s, matrix metalloproteinases(MMP)-3, MMP-13 and vascular endothelial growth factor (VEGF) were measured by real-time quantitative polymerase chain reaction and enzyme-linked immunosorbent assay (ELISA), respectively. Reactive oxygen species (ROS) generation was detected by fluorescent microscope and fluorescence microplate reader. Immunoprecipitation, Co-Immunoprecipitation and western blot were performed to examine the activity of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase and nuclear factor kappa B (NF-κB). Results: Exposure of FLSs to AOPPs upregulated the mRNA and protein expression of TNF-a, IL-1s, MMP-3, MMP-13 and VEGF in a concentration dependent manner. AOPPs treatment triggered ROS production in FLSs, which was significantly abolished by ROS scavenger N-acetyl-L-cysteine (NAC), superoxide dismutase (SOD), NADPH oxidase inhibitors diphenyleneiodonium (DPI) and apocynin. Challenged AOPPs induced phosphorylation of p47phox, triggered an interaction between p47phox, p22phox and gp91phox, and significantly upregulated expression of NADPH oxidase subunits p47phox, p22phox and gp91phox. IκB degradation and nuclear translocation of NF-κB p65 induced by AOPPs were significantly blocked by SOD, NAC, DPI and apocynin. Conclusions: These data indicate that AOPPs induce inflammatory response in FLSs is medicated through NADPH oxidase-dependent activation of NF-κB.

Relevant data including diagnosis and treatment model, operation situation, etc. are analyzed in our wound care center and compared with abroad based-on literature retrieval, to find out a suitable way for setting up wound care centers in our country. Wound care center with mode of multidisciplinary cooperation that is "specialist-oriented" was established in our hospital, mirroring the experience of foreign wound care centers. The investigation showed the demands of patients could be met, their psychological and economic pressures as well as social burden might be greatly relieved in our center. For an example, we achieved our purpose readily by setting up a clinical pathway of diabetic foot according to the features of our hospital. The model of multidisciplinary cooperation, with minor adjustments, may be applicable for wound care centers at home and abroad. Our wound care center will establish better system of diagnosis and treatment to conform to the situation in our country.

To investigate the influence of recombinant human growth hormone (rhGH) on body fluid compartments and water-sodium retention in severe burn patients.Thirty adult patients with severe burn were divided into treatment (T) and control (C) groups by block randomized design. Patients in both groups were subcutaneously injected with same amount of rhGH (12 IU/d) or isotonic saline during 7 - 21 post burn day (PBD). The total body water (TBW), intracellular water (ICW), extracellular water (ECW) were measured by bioelectrical impedance analysis (BIA) on 7, 14, 21 PBD. The 24 h urinary output of Na+ was determined by ion selective electrode method (ISE).There were no significant difference in levels of TBW, ICW, ECW and 24 h urinary output of Na+ between two groups on 7, 14, 21 PBD (P0.05). No difference in results was found between groups at different time points (P0.05). After the data were analyzed, the level of TBW (36 +/- 6 L), ICW (21 +/- 4 L) on 21 PBD were evidently lower than those on 7 PBD (38 +/- 6 L, 23 +/- 7 L, P0.01).The level of ICW and TBW in severe burn patients decreased along with the time. Proper dosage of rhGH has no significant effect on body fluid compartments and water-sodium retention.

To investigate the status of nutritional support therapy in moderate and severe burn patients.The burn patients with age over 16 y and burn area larger than 20% TBSA were enrolled in the retrospective study. According to length of stay, all patients were divided into the first period (271 cases, was from 1994 to 2001 year), and second period (273 cases, from 2002 to 2007 year), and they were subdivided into a (20% - 30% TBSA), b (31% - 50% TBSA), c (51% - 70% TBSA), d (larger than 70% TBSA) groups. The death rate and ratio of nutritional support therapy were compared. The change in albumin and other indices (including side-effects, complication, etc) were analyzed in each group in second period.There were similar in general conditions in both groups, however, some were obvious difference in two periods between nutritional support therapy rate (74.17% in first period vs 85.35% in second period, P0.01), and the ratio of parenteral nutrition to enteral nutrition (1.5:1.0 in first period, 1.0:1.5 in second period, P0.01). There were also significant differences in albumin levels among each group in second period on 4, 7, 14 days after burn (P0.05). In the second period, parenteral nutrition preparation was mostly delivered through central vein in the form of "all-in-one", among them 62 cases of positive catheter cultures were found. Enteral nutrition was delivered by oral route in 108 cases, naso-gastric or naso-enteric tubes with pump in 165 cases. 27 cases with severe gastrointestinal complications and 2 cases with inhalation pneumonia occurred.The ratio of parenteral nutrition and enteral nutrition in burn patients was higher in our ward than average level in China. There is still a long way ahead to optimize nutritional therapy.

Plasmodium falciparum DNA is detected with an assay modeled according to the reverse target capture assay described by Morrissey et al. [19] for the detection of Listeria cells. A poly(A)-tailed oligonucleotide (pWZ34), derived from the partial sequence of a 4-kb repetitive unit of P. falciparum, functions as a capture probe and the labelled 21-bp repetitive units specific for P. falciparum serve as a reporter probe. Both probes are complementary to non-overlapping regions of the target DNA and in the presence of high concentrations of chaotropic salts, hybridization efficiently takes place at relatively low temperatures (15 min, 37°C). The addition of poly(dT)-derivatized ferromagnetic beads allows the formation of A:T base pairing between the tailed beads and the tailed capture probe. Upon applying magnetic force, the target-capture-reporter-probe complex attached to the beads is removed from the reaction mixture, leaving the bulk of unreacted reporter molecules behind. Subsequent washings of the immobilized complex reduces the amount of non-specifically bound reporter probe. After elution of the complex from the beads a new cycle of capture, washing and release of the target-capture-reporter-probe complex is initiated by the additions of unused (dT)-tailed beads. After 3 cycles, the signal-to-noise ratio with 0.1 pg of P. falciparum DNA as a target was as high as 21–27, with a background of 8–10 cpm. The assay is unique in its speed, well suited for large sample numbers, and allows the manipulation of the background at will by simply increasing the number of capture rounds.

To investigate the effects of recombinant human growth hormone (rhGH) on the changes in serum insulin-like growth factor-I (IGF-I), IGF binding protein 3 (IGFBP-3) and blood sugar in severely burned patients, so as to validate the optimal time of rhGH administration.Forty severely burned patients were enrolled in the study and were randomly divided into control (C), treatment 1 (rhGH given from 7 - 9 PBD, T1) and treatment 2 (rhGH from 10 - 14 PBD, T2) groups. The dynamic changes in serum IGF-I, IGFBP-3 and blood sugar on the 1, 3, 5, 7, 10, 14 and 21 PBDs in all 3 groups of burn patients were determined, analyzed and compared with one another.The serum IGF-I, IGFBP-3 and blood sugar levels in T1 and T2 groups were higher than those in C group after the use of rhGH, especially the IGFBP-3 and blood sugar (P0.05). There was no difference of all the indices between T1 and T2 groups.It might be optimal to give rhGH to severely burned patients during 7-9 PBDs.

OBJECTIVE: To evaluate the therapeutic effect of the herbal plaster Tangshangxiaobagao on partial thickness burn wound. METHODS: A randomized controlled trial was conducted with two herbal plasters: Tangshang-xiaobagao and Jingwanhong in 57 hospitalized burn patients. Both the effect and safety of two herbal plasters were noted in patients with partial thickness burns. RESTULTS: In superficial second degree burns, the 7 d healing rate of both groups was (61.35+/-36.26)%and (51.21+/-37.24)% and the healing time (10.56+/-3.43)d and (11.98+/-4.13)d P0.05 respectively. While in deep second degree burns, the 14 d healing rate of both groups was (62.9+/-36.0) % and (53.9+/-32.2) % and the healing time (19.4+/-4.9)d and (21.5+/-5.5)d, respectively. Study group had lower VAS(visual analogue scale)score than control group. No obvious side effects were observed in study group. CONCLUSION: Tangshangxiaobagao is safe and may be an effective adjunct for treatment of partial thickness burn wounds.

BACKGROUND: Alendronate (ALE) is a conventional drug used to treat osteoporosis. Low-magnitude whole-body vibration (WBV) exercise has been developed as a potential treatment for osteoporosis. The aim of this study was to investigate whether low-magnitude WBV could enhance the protective effect of ALE on bone properties in ovariectomized rats. METHODS: A total of 128 Sprague-Dawley rats were randomly divided into five groups (SHAM, OVX+VEH, OVX+WBV, OVX + ALE, OVX+WBV+ALE). The level of WBV applied was 0.3 g at 45-55 Hz for 20 min/day, 5 day/week and for 3 months. ALE was administered in dose of 1 mg/Kg once a week. Every four weeks eight rats from each group were sacrificed and their blood and both tibiae were harvested. The expression of osteocalcin and CTX in serum was measured by enzyme-linked immunosorbent assay (ELISA) and the tibiae were subjected to metaphyseal three-point bending and μCT analysis. RESULTS: Osteocalcin rose after ovariectomy and was not appreciably changed by either alendronate or WBV alone or in combination. Alendronate treatment significantly prevented an increase in CTX. WBV alone treatment did not alter this effect. Compared with the OVX+WBV group, nearly all tested indices such as the BV/TV, TV apparent, Tb.N, Tb.Th, and Conn.D were higher in the OVX+ALE group at week 12.Compared with the OVX+WBV group, certain tested indices such as BV/TV, TV apparent, Tb.N, and Con.D, were higher in the OVX+WBV+ALE group at week 12. At week 12, tibiae treated with WBV+ALE exhibited a significantly higher Fmax compared to the OVX+VEH group, and a significant difference was also found in energy absorption between the OVX+WBV+ALE and OVX+VEH groups. CONCLUSIONS: Compared with the WBV, ALE was more effective at preventing bone loss and improved the trabecular architecture. However, WBV enhanced the effect of alendronate in ovariectomized rats by inducing further improvements in trabecular architecture.

The point mutation at nucleotide 323 within the dihydrofolate reductase-thymidylate synthase gene of Plasmodium falciparum , which distinguishes pyrimethamine-sensitive from drug-resistant isolates, can be discriminated by the polymerase chain reaction using mutation-specific primers. The technique makes use of the principle that short oligonucleotides with a perfect match at their 3′ ends, complementary to the mutation to be detected, will initiate the polymerization by Taq polymerase far more efficiently than primers with a single mismatch in this position. The Asn-108 codon was detected using a primer of 17 nucleotides with an adenosine at its 3′ end, the Thr-108 codon with a 14-mer primer ending with a cytosine and the Ser-108 codon with a 16-mer containing guanidine at the critical 3′ end. By selecting appropriate counterprimers, the size of the amplification products is either indicative of pyrimethamine-resistant parasites of the 7G8 type, or of pyrimethamine-sensitive parasites of the FCR-3 type or 3D7 type. The fragments obtained can be easily separated in a single lane after agarose gel electrophoresis. Coded P. falciparum DNA samples were typed unambiguously using these primers as were reconstituted parasitized blood samples stored as high salt lysates. Sensitivity, speed and specificity make this assay a realistic alternative to in vitro drug testing to monitor the resistance of P. falciparum to inhibitors of the dihydrofolate reductase.

The dihydrofolate reductase-thymidylate synthase (DHFR-TS) bifunctional complex from pyrimethamine-sensitive (3D7) and drug-resistant (HB3 and 7G8) clones from Plasmodium falciparum was purified to homogeneity. A modified sequence of purification steps with a 10-formylfolate affinity column at its center, allows the isolation of the enzyme complex with a 10-fold higher yield than previously reported, irrespective of the pyrimethamine resistance of the parasites. Titration of the homogeneous DHFR-TS complex with the inhibitor revealed a 500-fold lower affinity of the enzyme from clone 7G8 for the drug than found with the enzyme from clone 3D7. Direct comparison of the homogeneous enzyme preparations on SDS-PAGE revealed no difference in the molecular mass of the DHFR-TS from the 3 clones, nor could a reproducible difference be detected in the peptide patterns obtained after digesting the DHFR-TS complex with various proteases. The amplification of segments from the DHFR-TS coding region of the 3 clones and 7 isolates of P. falciparum by polymerase chain reaction resulted in fragments of the predicted length without any size heterogeneity. The DNA sequence of the DHFR coding region from FCR-3, 3D7, HB3 and 7G8 differs in a total of 4 nucleotides. One point mutation changes amino acid residue 108 from threonine (FCR-3) or serine (3D7) to asparagine (HB3 and 7G8). The presence of asparagine-108 appears to be the molecular basis of pyrimethamine resistance of HB3 and 7G8. The degree of resistance is associated with a point mutation affecting the codon for amino acid 51 in 7G8.

We propose here the formation by molluscan and notochord muscles in the catch state of three-dimensional, entangled network structures composed of bent and sometimes entwined paramyosin thick filaments including myosin intermediate filaments and disordered actin thin filaments; in the relaxed state the three forms lie in parallel. The intact forms of bivalve (Andonta pacifica, Heude) muscle paramyosin are those of 120 and 95 kDa (β-paramyosin). The 102 kDa form (α-paramyosin) is the proteinase cleavage product of 120 kDa paramyosin. Paramyosin could be phosphorylated in vitro by cyclic AMP-dependent protein kinase. The amino acid phosphorylated was at the serine residue. Paramyosin from muscles treated with acetylcholine (catch state) was phosphorylated to a greater extent than that of untreated muscles (normal state) and even more so in the case of serotonin-treated muscles (relaxed state). Actin markedly inhibited the phosphorylation of paramyosin in vitro.

The rate and yield of oxidative renaturation of reduced RNase A has been studied as a function of [-S-S-]/[-SH]. The principal conclusion of these studies is that rates and yields of oxidative renaturation are strongly dependent on the low mol. wt disulfide/thiol ratio. The relationships are complex and do not parallel the redox potential of the system. The present results are consistent with earlier findings on other proteins, and lead us to believe that the above conclusion is general. Kinetic studies of oxidative renaturation should recognize and account for the dependence of reaction rate and extent on the disulfide/thiol ratio. This ratio can change substantially over the course of a reaction, either due to stoichiometric transfer of disulfide to protein, and/or adventitious air oxidation of thiols. Failure to account for changes in the disulfide/thiol ratio may compromise the interpretation of such experiments.