[The application of improved Glasgow coma scale score of 15 as switching point for invasive noninvasive mechanical ventilation in treatment of severe respiratory failure in chronic obstructive pulmonary disease].

OBJECTIVE: To estimate the feasibility and the efficacy of early extubation and the sequential non invasive mechanical ventilation (MV) in severe respiratory failure of chronic obstructive pulmonary disease (COPD) with the improved Glasgow coma scale (GCS) score of 15 as the switching point. METHODS: By a prospective control study, 20 patients with COPD and respiratory failure who had undergone endotracheal intubation and MV from March 2007 to November 2009 were enrolled as treatment group. Invasive MV with synchronous intermittent mandatory ventilation and pressure support ventilation (SIMV+PSV) pattern were given to these patients. When the period of "improved GCS score of 15 standard" window period appeared and being kept for 2 hours, endotracheal tube was extubated, and nasal mask with PSV+positive end expiratory pressure (PEEP) was used, followed by gradual decrease of the level of pressure support till weaning of MV. Nineteen patients who were treated with MV with ordinary way of weaning from March 2005 to March 2007 served as the control group. Prior to the MV, the ventilation and oxygenation index , the length of invasive MV, total MV time, total hospital stay, re intubation and ventilator associated pneumonia (VAP) occurred in the number of cases were observed and compared between two groups. RESULTS: There was no significant difference in the ventilation and oxygenation index prior to the MV. Compared with control group, in treatment group, the length of invasive ventilation (days: 3.2±1.1 vs. 10.5±3.2), the total duration of MV (days: 4.8±2.5 vs. 10.5±3.2), the length of hospital stay (days: 17±3 vs. 22±7) were significantly shorter (all P<0.01), and the incidence of VAP was significantly lower (cases: 0 vs. 5, P<0.01), while the number of re intubation was slightly higher but without statistical significance (cases: 3 vs. 1, P>0.05). CONCLUSION: The application of improved GCS score of 15 as the switching point with 2 hours as window period for early extubation and non invasive nasal mask ventilation can significantly improve the therapeutic effect in patients with severe respiratory failure in COPD.

[Changes in serum contents of inflammatory mediators after tetramethylene-disulfo-tetramine poisoning and clinical study of a new treatment regime for the poisoning].

OBJECTIVE: To study the changes in serum contents of beta-endorphin (beta-EP), endothelins (ET), nitric oxide (NO) and tumor necrosis factor (TNF) after acute tetramethylene-disulfo-tetramine (TDT) poisoning and therapeutic effect of a new treatment regime. METHODS: (1) Forty-eight patients with tetramethylene-disulfo-tetramine poisoning (experiment group) were enrolled in this study. The serum levels of beta-EP, ET, NO and TNF were measured upon hospitalization and 1, 3, 5, 7, 9, 11, 13, 15, 17 and 19 days after poisoning, respectively, and compared with those of 30 healthy individuals (control group B). (2) They were treated with the improved regime and compared with patients treated with the conventional regime designated as control group A. RESULTS: (1) In 48 patients treated with improved regime, 45 were cured and 3 died. (2) The serum levels of beta-EP, ET, NO and TNF from 45 patients who were cured were significantly higher at hospitalization compared with those of healthy individuals, with the peak values appeared on day 1 after poisoning in the mild, moderate and severe groups. Beta-EP levels returned to normal range on days 9, 13 and 17 after poisoning respectively in the mild, moderate and severe groups. ET levels returned to normal range on days 7, 13 and 15 after poisoning respectively in the mild, moderate and severe groups. NO levels returned to normal range on days 7, 11 and 11 after poisoning respectively in the mild, moderate and severe groups. TNF levels returned to normal range on days 9, 11 and 17 after poisoning respectively in the mild, moderate and severe groups. (3) The serum levels of beta-EP, ET, NO and TNF in 3 non-survivors were very high at hospitalization and continued to increase in the course of treatment. (4) The cumulative doses of diazepam and Phenobarbital, and the eclampsia time were significantly less in the experiment group than those of control group A. CONCLUSION: (1) The serum levels of beta-EP, ET, NO and TNF are correlated with the severity of tetramethylene-disulfo-tetramine poisoning and general conditions of the patients. (2) When the serum levels of beta-EP, ET, NO and TNF decrease gradually in the course of treatment, prognosis is better. On the contrary, the prognosis is poor when their levels increase gradually. (3) Measures to decrease levels of beta-EP, ET, NO and TNF result in a better prognosis of patients with tetramethylene-disulfo-tetramine poisoning. (4) The improved regime can be considered a better therapeutic strategy in tetramethylene-disulfo-tetramine poisoning.

Toxicity of two type II ribosome-inactivating proteins (cinnamomin and ricin) to domestic silkworm larvae.

Cinnamomin and ricin are two type II ribosome-inactivating proteins. They exhibited a different toxicity to domestic silkworm (Bombyx mori) larvae by oral feeding bioassay. The LC50 of ricin to the silkworm larvae at third instar was much lower than that of cinnamomin. When the isolated 80S ribosome from domestic silkworm pupae was treated separately with the reduced cinnamomin or the reduced ricin, a specific RNA fragment (R-fragment) was produced as characterized by 8 M urea-denatured polyacrylamide gel (3.5%) electrophoresis. The purified A-chains of both cinnamomin and ricin showed a slightly different RNA N-glycosidase activity to the domestic silkworm pupal ribosome. It was proposed that the difference of their toxicity to domestic silkworm larvae was not related to their A-chains but to the properties of their B-chains. It was also found that the vomit obtained from the midgut of domestic silkworm larvae could hydrolyze these two proteins apparently to a similar extent.

Isolation and enzymatic characterization of lamjapin, the first ribosome-inactivating protein from cryptogamic algal plant (Laminaria japonica A).

Lamjapin, a novel type Iota ribosome-inactivating protein, has been isolated from kelp (Laminaria japonica A), a marine alga. This protein has been extensively purified through multiple chromatography columns. With a molecular mass of approximately 36 kDa, lamjapin is slightly larger than the other known single-chain ribosome-inactivating proteins from the higher plants. Lamjapin can inhibit protein synthesis in rabbit reticulocyte lysate with an IC50 of 0.69 nm. It can depurinate at multiple sites of RNA in rat ribosome and produce the diagnostic R-fragment and three additional larger fragments after the aniline reaction. Lamjapin can deadenylate specifically at the site A20 of the synthetic oligoribonucleotide (35-mer) substrate that mimics the sarcin/ricin domain (SRD) of rat ribosomal 28S RNA. However, it cannot hydrolyze the N-C glycosidic bond of guanosine, cytidine or uridine at the corresponding site of the A20 of three mutant SRD RNAs. Lamjapin exhibits the same base and position requirement as the ribosome-inactivating proteins from higher plants. We conclude that lamjapin is an RNA N-glycosidase that belongs to the ribosome-inactivating protein family. This study reports for the first time that ribosome-inactivating protein exists in the lower cryptogamic algal plant.

Purification of alpha-sarcin and an antifungal protein from mold (Aspergillus giganteus) by chitin affinity chromatography.

A simple method for preparation of alpha-sarcin and an antifungal protein (AFP) from mold (Aspergillus giganteus MDH 18894) has been developed. alpha-Sarcin and AFP were purified simultaneously by chitin affinity column chromatography and gel filtration. By this method, 4.5 mg of pure alpha-sarcin and 6.9 mg of pure AFP were obtained from 2 liters of culture medium. Compared with other purification methods such as ion-exchange column chromatography, this procedure was very simple and specific. The purified alpha-sarcin and AFP were homogeneous as characterized by SDS-polyacrylamide gel electrophoresis. Both alpha-sarcin and AFP exhibited the binding activity to generated chitin. Soluble glycochitin decreased the intensity of fluorescence of alpha-sarcin and made the lambda(em)m shift from 340 to 347 nm. Titration of alpha-sarcin with N-bromosuccinimide under native conditions revealed that two tryptophans (Trps) were all located in the core part of alpha-sarcin molecule. This indicated that Trps were not involved in the binding of alpha-sarcin to chitin. Glycochitin in the culture medium increased the expression of alpha-sarcin, while it had no effect on the expression of AFP. Unlike other ligands such as Cibacron blue for the affinity purification of alpha-sarcin and AFP, glycochitin increased the nuclease activity of alpha-sarcin.

Cinnamomin, a type II ribosome-inactivating protein, is a storage protein in the seed of the camphor tree (Cinnamomum camphora).

Cinnamomin is a novel type II ribosome-inactivating protein (RIP) isolated in our laboratory from the seed of the camphor tree (Cinnamomum camphora). In this paper the physiological role it plays in the plant cell was studied. Northern and Western blotting revealed that cinnamomin was expressed specifically in cotyledons. It accumulated in large amounts simultaneously with other proteins at the post-stages of seed development. Cinnamomin degraded rapidly during the early stages of seed germination. Endopeptidase was proved to play an important role in the degradation of cinnamomin. Western blotting of total proteins from the protein body with antibodies against cinnamomin demonstrated that it only existed in this specific cellular organelle as a storage protein. The similar properties of cinnamomin and other seed storage proteins of dicotyledons were compared. We conclude that cinnamomin is a special storage protein in the seed of C. camphora.

Studies of three genes encoding Cinnamomin (a type II RIP) isolated from the seeds of camphor tree and their expression patterns.

Cinnamomin, which has three isoforms, is a type II ribosome-inactivating protein (RIP) purified from the mature seeds of camphor tree (Cinnamomum camphora). In a previous study, an incomplete cDNA that encoded the A- and B-chain of Cinnamomin but lacked signal peptide sequence was cloned. In the present paper, its full-length cDNA was obtained by 5' rapid amplification of cDNA ends (5'RACE). Subsequently, polymerase chain reaction (PCR) amplification of its genomic DNA was performed. Unexpectedly, sequence analysis of the PCR products revealed three cinnamomin genes with >98.0% sequence identity. One of them corresponded to the published cDNA and was designated as cinnamomin I, whereas the other two genes were named as cinnamomin II and cinnamomin III, respectively. RT-PCR amplification of the cDNAs of cinnamomin II and III manifested that these two genes were functional. The three genes have no intron. Three Cinnamomin precursors that were inferred from the cDNA sequence of three cinnamomin genes exhibited relatively high sequence homology with other type II RIPs. Northern blot analysis demonstrated that the cinnamomin genes only expressed in cotyledons of C. camphora seeds and the acmes of expression emerged at 75-90 DAF when seeds were close to maturity. It is proposed that the three cinnamomin genes may encode three isoforms of Cinnamomin. The physiological function of Cinnamomin in C. camphora seeds is briefly discussed.