As a novel post-translational modification(PTM),lysine 2-hydroxyisobutyrylation(Khib)is considered to regulate gene transcriptional activities in eukaryotic cells;however,the functions of Khib-modified proteins in plants remain unknown.Here,we report that Khib is an evolutionarily-conserved PTM in wheat and its progenitors.A total of 3348 Khib sites on 1074 proteins are iden-tified in common wheat(Triticum aestivum L.)by using affinity purification and mass spectroscopy of 2-hydroxyisobutyrylome.Bioinformatic data indicate that Khib-modified proteins participate in a wide variety of biological and metabolic pathways.Immunoprecipitation confirms that Khib-modified proteins are present endogenously.A comparison of Khib and other main PTMs shows that Khib-modified proteins are simultaneously modified by multiple PTMs.Using mutagenesis experiments and co-immunoprecipitation assays,we demonstrate that Khib on K206 of phospho-glycerate kinase(PGK)is a key regulatory modification for its enzymatic activity,and mutation on K206 affects the interactions of PGK with its substrates.Furthermore,Khib modification of low-molecular-weight proteins is a response to the deacetylase inhibitors nicotinamide and tricho-statin.This study provides evidence to promote our current understanding of Khib in wheat plants,including the cooperation between Khib and its metabolic regulation.

AIM: To extract and isolate polysaccharide from Poacynum Hendersonii leaves,determine its content and analyze the monosaccharide composition.METHODS: Poacynum Hendersonii leaves was extracted with hot water,crude polysaccharide was precipitated with ethanol,deproteinated according to Sevage method,coloured with acticarbon.Then of polysaccharide contents were measured by anthrone-H_2SO_4 colorimetry at the wavelength of 620 nm.The monosaccharide composition was determined by HPCE.RESULTS: The polysaccharide content was 0.97% of leaf weight,and Gal,Ara,and Man contents were three higher monosaccharides.CONCLUSION : The method is easy to carry out the baseline resolution in HPCE and has highly sensitivity.

The purpose of this study was to investigate bacterial translocation and change in intestinal permeability in patients after abdominal surgery. Sixty-three patients undergoing elective abdominal surgery were enrolled in the study. Blood samples were collected prior to operation and 2, 24, 48 h after surgery for bacterial culture, microbial DNA extraction, plasma D-lactate and endotoxin measurement. PCR analysis was performed after DNA extraction, with beta-lactosidase gene of E. coli and 16S rRNA gene as target genes. All patients were observed for a period of 30 days for infectious complications. Our results showed that no bacterial DNA was detected before surgery, but after operation it was found in 12 patients (19.0%). Bacterial DNA was detected in 41.7% (10/24) of SIRS patients and 5.1% (2/39) of non-SIRS patients (P<0.01). About 83.3% of PCR-positive patients developed systemic inflammatory response syndrome (SIRS), but only 27.5% of PCR-negative patients did so (P<0.01). Two thirds of PCR-positive patients developed infectious complications, while none of PCR-negative patients did (P<0.01). The blood culture was positive only in 3 patients (4.8%), who were all PCR-positive. E. coli DNA was found in 66.7% of the PCR-positive patients. The plasma levels of D-lactate and endotoxin were elevated significantly 2, 24 and 48 h after operation in PCR-positive patients, with a significant positive correlation found between them (r=0.91, P<0.01). It is concluded that increased intestinal permeability was closely related with bacterial translocation. Intestinal bacterial translocation (most commonly E. coli) might occur at early stage (2 h) after abdominal surgery. Postoperative SIRS and infection might bear a close relationship with bacterial translocation.

sely related with bacterial translocation. Intestinal bacterial translocation (most commonly E. coli) might occur at early stage (2 h) after ab-dominal surgery. Postoperative SIRS and infection might bear a close relationship with bacterial translocation.

Objective To investigate the effects of carbachol on intestinal inflammation and mucosal blood flow after gut ischemia-repedusion(I/R) in rat. Method A jejunal sac was formed in Wistar rats. The superior mesenteric artery (SMA) was occluded for 45 mi-nutes followed by 240 minutes of reperfusion. Animals were random divided into three groups: sham operation, L/R + saline injection (I/R + NS) and I/R + carbachol injection (0.1mg/kg, I/R + Ca). Immediately after occluded of SAM blood flow, either 0.1mg/kg of carba-chol or same account of 0.9% saline was injected into the jejunal sac. The pathological injury was observed with HE staining. The activity of DAO and content of TNF-α in intestinal mucosa tissue were determined. Mucosal blood flow was measured by laser Doppler. All measure-ments were done at 0 min, 30 min, 60 min, 120 min, and 240 min after reperfusion. Result In I/R group the activity of DAO in intestinal mucosa and mucosal blood flow deceased, meanwhile the content of TNF-α gut tissue was dramatically increased than those in sham operation (P<0.01). Severe pathological changes were observed in intestinal mucosa. After injection of carbachol, the activity of DAO and mucosal blood flow increased (P<0.01), but the content of TNF-α in intestinal mucosa were dramatically decreased (P<0.01), compared with those in I/R group. Conclusion Administration of carbachol protects intestinal ischemia-reperfusion injury by attenuating intestinal mucosa inflammation and increasing gut mueosal blood flow.

Aim To establish a rapid and sensitive diagnostic markerfor adjusting gut absorptive capacity after trauma. Methods A mi-cromethod for D-xylose test with phloroglucinal was established according toEberts with modification. The standard curves were repeated for fivetimes. Then the blood contents of fasting and orally taking 0. 5 g/kgD-xylose at 2nd and 4th hour were measured and the blood D-xylose con-tents of seven refilling rats with ischemia of intestine were mea-sured. Results The maximal absorption spectra was read at 554 nm byscanming of DU-7 Beckman. Good linearity of the D-xylose standard curvewas found with the range of 0 to 4 mmol/L(r=0. 9979 ±0. 0017) . Theblood content of rats at 2nd hour after orally taking D-xylose was ( 154 ± 6)mg/L amd that at 4th hour was decreased to (87 ± 11) mg/L. And theblood content at 2nd and 4th day after ischemic refilling was (162 ± 5)mg/L and (80 ± 8) mg/L respectively. The variation coefficients within andamong batches were 1.98% (n=6) and 3.10% (n=6), respective-ly. And D-xylose recovery rates were from 97.2% to 104. 3%. ConclusionMicromethod for D-xylose test appears to be simple, rapid and sensitive,and is an available index for estimating intestinal absorption after severetrauma.

OBJECTIVETo describe the methods which were used to develop collagen-based materials for wound dressing.

METHODSFresh frozen bovine tendon was treated with 0.05 mol/L acetic acid at pH 3.2 for 48-72 hours, homogenized, filtered, mixed with 8% chondroitin sulphate, for creating a deaerated 1.5%-2.5% collagen solution. The solution was lyophilized in either a pre-frozen or non-pre-frozen mould. The collagen sponge was then cross-linked with 0.25% glutaraldehyde for 24 hours. Three other types of wound dressings were developed using a similar method: collagen membrane with a polyurethane membrane onlay, polyurethane-coated collagen membrane and collagen membrane on gauze.

RESULTSIt was demonstrated that the use of frozen bovine tendon was stable, and that the prepared collagen sponge contained pores of 50-400 microm in diameter.

CONCLUSIONSCollagen could be used as wound dressing.

Amino Acids ; analysis ; Animals ; Biological Dressings ; Cattle ; Collagen ; analysis ; chemistry ; isolation & purification ; Freeze Drying ; Polyurethanes

OBJECTIVETo investigate the proliferative effect of keratinocyte growth factor (KGF-2) on human adult keratinocytes.

METHODSThe standard medium was keratinocyte growth medium without bovine pituitary extract (BPE), hydrocortisone or epidermal growth factor (EGF). Keratinocytes from a 48-year-old subject were cultured and seeded on dishes with standard medium of EGF in cell density of 2 x 10(4)/32 mm(2). After 24 hours, the medium was replaced by the standard medium with 0, 4, 16, 125 and 500 ng/ml KGF-2, respectively. The standard medium with EGF was used as the positive control and the standard medium without EGF or KGF-2 was used as the negative controls. The growth of keratinocytes was monitored by 3-(4,5-dimethythiazol-2-yl)-2,5 dipheyl tetrazolium bromide (MTT) assay and by photographs on days 3, 5 and 7, respectively.

RESULTSKGF-2 in concentrations of 4-500 ng/ml showed a significant proliferative effect on days 5 and 7 as compared with that of the negative controls (P < 0.01). On day 3 the cells were proliferated to 1.5-2.5-fold, on day 5 to 3-5-fold and on day 7 to 3-12-fold in KGF-2 medium as that of the negative controls. The optimal response occurred when the concentration of KGF-2 was 125 ng/ml on day 7. Cell proliferation was also consistently higher in all KGF-2 concentrations as compared with that of the positive controls.

CONCLUSIONSKGF-2 has significant effects on the proliferation of adult keratinocytes, which are more effective than that of EGF. This study supports KGF-2 can improve the healing of chronic wounds in adults in clinic.

Analysis of Variance ; Cell Division ; drug effects ; physiology ; Cells, Cultured ; Culture Media, Conditioned ; Epidermal Growth Factor ; pharmacology ; Fibroblast Growth Factor 7 ; Fibroblast Growth Factors ; pharmacology ; Humans ; Keratinocytes ; drug effects ; physiology ; Middle Aged ; Probability ; Reference Values ; Sensitivity and Specificity

OBJECTIVETo obtain information about the quality of scars of healed venous leg ulcers compared with intact skin on the opposite leg by using high-frequency ultrasound.

METHODSTwenty-eight patients (16 women, 12 men, aged 31 - 89 years) whose venous ulcers had healed and scars formed were included in this study. The echogenicities of scars were measured with a 20 MHz high-frequency ultrasound Dermascan. The thickness of epidermis and dermis was assessed and the number of low echogenic pixels (LEPs) in the papillary dermis and reticular dermis were counted using image analysis software.

RESULTSThe average epidermal thickness of the scars after 1 week to 20 years of healing was significantly increased compared to those of the control (P < 0.01), whereas the average dermal thickness of scars after healing was significantly decreased compared to the control (P < 0.01). The numbers of LEPs and the distributions of LEPs between scars and controls had no statistically significant differences. There were no correlations among scar echogenicities, age of healed venous ulcers, initial ulcer areas, age of venous ulcers or age of patients. In the control skin samples, the young group aged 31 - 69 years had fewer LEPs than did the elderly group aged 70 - 89 years.

CONCLUSIONOur study demonstrates that after the healing of venous leg ulcers, there are significant differences in the thickness of the epidermis and dermis, but no significant alterations in water content and distribution in the dermis when compared to the controls.

Adult ; Age Factors ; Aged ; Aged, 80 and over ; Cicatrix ; diagnostic imaging ; Female ; Humans ; Male ; Middle Aged ; Skin ; diagnostic imaging ; Time Factors ; Ultrasonography ; Varicose Ulcer ; diagnostic imaging

Objective To study the injury factors, pathogenic process and clinical features of delay two-phase multiple organ dysfunction syndrome (MODS) in severe burned patients and to replicate a standardized animal model that would accurately imitate the clinical features of MODS.Methods Forty-five human patients with burn size larger than 30% total body surface area (TBSA) were analyzed. All of them underwent severe burn shock in early stage and sepsis in late stage. Thirty-two goats were randomly divided into three groups: 1) hemorrhagic shock (group H, n=6); 2) endotoxemia (group E, n=6); and 3) hemorrhagic shock plus endotoxemia (group M, n=20). Hemorrhagic shock was produced according to the method of Wigger (6.7 kPa for an hour, 1 kPa=7.5 mmHg). Endotoxin (E. coli O111 B4) was given via the portal vein 24 hours after the resuscitation of hemorrhagic shock, in a dose of 30 ng/kg/min for 5 consecutive days. During the observation period of 10 days, all animals were hemodynamically monitored, given standard metabolic support and due cardiac and pulmonary support according to human intensive care.Results All the patients showed burn shock at 1-3 days and hyperdynamic circulation, hypermetabolism and systemic inflammatory responses over two weeks post-injury. Thirteen cases were found to develop MODS according to the prevailing diagnostic criteria, and 10 of them died with a mortality of 77%. Eighteen animals died in group M with a mortality of 90%, 12 of the 18 developed MODS, with overall incidence of 60%. Most animals in group M showed changes similar to that observed in human cases. The experimentation proved that in the pathogenic process of MODS, there was a two-hit phenomenon in the dvelopment of the syndrome. To prevent the development of MODS, it therefore was imperative to blunt the first hit or the second hit, so that an excessive inflammatory response was alleviated. This postulation has been verified in the treatment of extensive burns. Two patients with burn extent reaching 100% TBSA survived with only mild acute respiratory distress syndrome (ARDS) and renal dysfunction after comprehensive treatment of burn shock, including adequate fluid resuscitation, drugs to remove oxygen free radicals, rapid restoration of pHi, and early extensive excision of burn eschars.Conclusion Both in human patients or animal experimentation, the typical delay two-phase MODS is shown to be produced by two successive insults in the forms of hypovolemic shock and sepsis. This postulation is helpful in formulating the prevention and treatment modality of MODS.