Objective:To explore the influence of intensive analgesia on the incidence of post-traumatic stress disorder (PTSD) in acute trauma patients, and to develop new ideas for the prevention and treatment of PTSD.Methods:From January 2018 to November 2019, a prospective study was conducted on trauma patients who visited the Emergency Center of Affiliated Hospital of Xuzhou Medical University and met the enrollment criteria. The patients were divided into the intensive analgesia group (< 4) and non-intensive analgesia group (≥ 4) according to the mean pain score in 30 days. The epidemiological data, trauma-related parameters, analgesic schemes, VAS score, PCL-5 score, HADS score and incidence of PTSD of enrolled patients were collected. Appropriate statistical methods were used to analyze differences among the indicators between the two groups.Results:Eighty-four acute trauma cases were included in the study, 39 cases in the intensive analgesia group and 45 in the non-intensive analgesia group. There was no significant difference in baseline data between the two groups (all P>0.05). The incidence rate of PTSD and PCL-5 score of patients in the intensive analgesia group were all significantly lower than those in the non-intensive analgesia group in 1 month after the trauma (all P< 0.05). The HADS anxiety and depression scores of patients in the intensive analgesic group were significantly lower than those in the non-intensive analgesic group (all P< 0.05). All the analgesics were converted into the dosage of dezocine for comparison. The total dosage of analgesics (dezocine) used in patients of the intensive analgesia group was significantly higher than that in the non-intensive analgesia group within 30 days after injury ( P< 0.05). Conclusion:In the acute trauma patients, intensive analgesia after trauma can significantly reduce the incidence of PTSD as well as improve anxiety and depression symptoms.

AIMTo study the expression of vascular endothelial growth factor (VEGF) in U937 foam cells and the inhibitory effect of salvianolic acid B and Ginkgo biloba extract in vitro.

METHODSU937 cells were incubated with 80 mg.L-1 oxidized low density lipoprotein (OX-LDL) for 48 h and a macrophage-derived foam cell model was established. The VEGF concentration in the media was determined by ELISA; the VEGF protein expression in cells was measured with immunohistochemistry; the VEGF mRNA level in cells was measured by in situ hybridization; the positive ratio detected by a morphometrical analysis system was used as the amount of the VEGF protein expression and the mRNA level.

RESULTSAfter U937 cells were incubated with OX-LDL, VEGF expression level increased greatly both in the cells and in the media. Salvianolic acid B and Ginkgo biloba extract were shown to remarkably inhibit the increase of VEGF. After treated with 10 micrograms/L-1 salvianolic acid B and Ginkgo biloba extract, the VEGF protein concentration in the media and positive ratio in the cells decreased compared with foam cells. After treated with 10 micrograms.L-1 salvianolic acid B and 100 micrograms.L-1 Ginkgo biloba extract, the VEGF mRNA level decreased measured by in situ hybridization.

CONCLUSIONA high VEGF expression level was determined in U937 foam cells. Salvianolic acid B and Ginkgo biloba extract were found to inhibit VEGF expression significantly in U937 foam cells in vitro.

Benzofurans ; pharmacology ; Foam Cells ; drug effects ; metabolism ; Gene Expression ; drug effects ; Ginkgo biloba ; chemistry ; Humans ; Plant Extracts ; isolation & purification ; pharmacology ; RNA, Messenger ; biosynthesis ; genetics ; U937 Cells ; Vascular Endothelial Growth Factor A ; biosynthesis ; genetics

This study is to investigate the protein and mRNA expressions of pro-inflammatory and anti-inflammatory cytokines in U937 foam cells and effects of Ginkgo biloba extract (GbE) on the cytokines. U937 cells were cultured with different concentrations of GbE (0.1, 1, and 10 microg x L(-1)), and stimulated by 100 mg x L(-1) oxidized low density lipoprotein (ox-LDL) for 24 h. The expressions of interleukin-1beta (IL-1beta), tumor necrosis factor-alpha (TNF-alpha) and interleukin-10 (IL-10) in culture solution were detected by enzyme-linked immunosorbant assay (ELISA) and reverse transcriptase polymerase chain reaction (RT-PCR). The results showed that incubated with 100 mg x L(-1) ox-LDL for 24 h, the U937 cells became foam cells, the protein or mRNA expressions of IL-1beta, TNF-alpha, IL-10, and its receptor IL-10R in U937 foam cells were higher markedly than those in normal U937 cells. When the cells were pretreated with GbE (0.1, 1, and 10 microg x L(-1)), the increases of IL-1beta and TNF-alpha in U937 foam cells were remarkably inhibited, but IL-10 expression increased greatly. Especially when cells were pretreated with 10 microg x L(-1) GbE, the protein and mRNA expressions of IL-1beta and TNF-alpha were markedly lower than those in U937 foam cells. The protein expression of IL-10 and mRNA expressions of IL-10 and its receptor IL-10R were markedly higher than those in U937 foam cells. GbE inhibited production of pro-inflammatory cytokines IL-1beta and TNF-alpha, but up-regulated the production of anti-inflammatory cytokine IL-10 and its receptor IL-10R in U937 foam cells, which might be related with its anti-atherosclerotic actions.
Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Foam Cells ; metabolism ; Ginkgo biloba ; chemistry ; Humans ; Interleukin-10 ; biosynthesis ; genetics ; Interleukin-1beta ; biosynthesis ; genetics ; Lipoproteins, LDL ; Plants, Medicinal ; chemistry ; RNA, Messenger ; metabolism ; Receptors, Interleukin-10 ; biosynthesis ; genetics ; Tumor Necrosis Factor-alpha ; biosynthesis ; genetics ; U937 Cells

OBJECTIVETo compare the ingredients of essential oils of Curcuma wenyujin extracted by supercritical-CO2 fluid extraction and by steam distillation.

METHODGC-MS was applied in this experiment.

RESULTThe ingredients and physical and chemical properties of essential oils of C. wenyujin extracted by supercritical-CO2 fluid extraction and by steam distillation are similar.

CONCLUSIONSupercritical-CO2 fluid extraction is better than steam distillation in extraction time, power consumption, recovery and purity.

Carbon Dioxide ; Chromatography, Supercritical Fluid ; methods ; Curcuma ; chemistry ; Oils, Volatile ; chemistry ; isolation & purification ; Plants, Medicinal ; chemistry ; Rhizome ; chemistry ; Volatilization

The hybrid xylanase TB was constructed by the substitution of the N-terminus segment of the Streptomyces olivaceoviridis xylanase XYNB with corresponding region of Thermomonosporafusca xylanase TfxA. The hybrid gene tb, encoding the TB, was correctly expressed in Escherichia coli BL21 and Pichia pastoris GS115. TB was purified and its enzymatic properties were determined. The results revealed that the optimal temperature and optimal pH of TB were at 70 degrees C and 6.0, which have been obviously improved compared with those of XYNB. The thermostability of TB were all about six-fold of XYNB's after incubating the properly diluted enzyme solutions at 80 degrees C and 90 degrees C for 3min, respectively. The pH stability of TB was 5 to approximately 9, which was narrower than that of XYNB. Still, TB remains a high specific activity as XYNB does. Analysis of a homology modeling and sequence similarity were used to reveal the factors influencing the enzymatic properties of TB and the discussion for the relationship between structure and function of xylanase was given.
Amino Acid Sequence ; Base Sequence ; Desulfurococcaceae ; enzymology ; genetics ; Endo-1,4-beta Xylanases ; genetics ; metabolism ; Enzyme Stability ; Escherichia coli ; enzymology ; genetics ; Hot Temperature ; Molecular Sequence Data ; Pichia ; enzymology ; genetics ; Protein Engineering ; methods ; Recombinant Fusion Proteins ; genetics ; metabolism ; Streptomyces ; enzymology ; genetics ; Structure-Activity Relationship

BACKGROUND:The vascular endothelial growth factor (VEGF) plays an important role in the development and formation of blood vessels.Up to now,there are few reports about the treatment of postoperative complications of vascular anastomosis surgery by mcrosutures with VEGF in China.OBJECTIVE:To synthesiize microsutures with VEGF and to evaluate its effect in revascularization following small vessel anastomosis.METHODS:The method of emulsification-diffusion was use to produce biodegradable polymer polylactic acid/glycolic acid (PLGA) copolymer microparticles containing VEGF,and then,the microparticles were added into microsutures to prepare microsutures with VEGF.Ninety Sprague-Dawley rats were enrolled to make animal models of caudal artery anastomosis using microsutures with VEGF in experimental group and microsutures alone in control group.Complications and VEGF level in the peripheral blood were detected and hematoxylin-eosin staining at the anastomotic site was performed at 2,12 hours,1,3,7 days after anastomosis.RESULTS AND CONCLUSION:(1) Postoperative complications:The postoperative incidence of skin necrosis was significantly lower in the experimental group than the control group (P ＜ 0.05).(2) VEGF level:Compared with the control group,the peripheral blood VEGF level was significantly higher in the experimental group at each time point after operation (P ＜ 0.05).(3) Hematoxylin-eosin staining:In the experimental group,proliferated endothelial cells were seen near the anastomotic site at 1 day after anastomosis;there were a large number of proliferated endothelial calls and subcutaneous tissues covering the sutures completely at 3 days after anastomosis;and endothelial cells and internal elastic lamina were completely repaired,smooth muscle cells proliferated further,and the outer membrane returned to normal at 1 week after anastomosis.In the control group,cell degeneration and necrosis were seen near the anastomotic suture,and only adventitial cells infiltrated and exhibited a traumatic proliferative response at 1 day after anastomosis;neonatal endothelial cells appeared in the exfoliated area of the endothelial cells,grew and migrated,and there was a few endothelial cells covering the anastomotic site at 3 days after anastomosis;and newborn endothelial cells got over the anastomotic crack and covered the suture.To conclude,microsutures with sustained-release VEGF microparticles can promote endothelial cell regeneration in rats at the anastomotic site.

OBJECTIVETo investigate the effect of membrane type-1 matrix metalloproteinase (MTI-MMP) on the invasive potential of breast cancer cell and analyze its mechanisms.

METHODSAfter treatment of breast cancer MDA-MB-453 cell line with concanavalin A ( ConA, 20 microg/ml) for 24 h, MT1-MMP protein was detected in cancer cells by Western analysis and immunocytochemistry. MDA-MB-453 cells were cultured with exogenous latent proMMP-2 and MMP-2 activity was analyzed by gelatin zymography. The invasive potential of the tumor cells was measured with a membrane invasion culture system. Cancer cells of the cell line were divided into four groups: the control group treated by neither reagent, group ConA was only treated by ConA, group MMP-2 was treated only by MMP-2, and group ConA + MMP-2 was treated by both ConA and MMP-2. RESULTS The expression of MTI-MMP protein could be detected in groups ConA and ConA + MMP-2, but nothing was detected in control and group MMP-2. There was only 72 000 precursor form of MMP-2 in group MMP-2 and there were both 72 000 precursor form and 64 000 active enzyme form of MMP-2 in group ConA + MMP-2, but there was no forms of MMP-2 in the other two groups detected by gelatin zymography. The largest amount of cells penetrated through Matrigel was observed in group ConA + MMP-2 than in the other three groups.

CONCLUSIONMTI-MMP can remarkably promote the invasive potential of breast cancer cells mainly through its ability of activating latent proMMP-2 to degrade

Blotting, Northern ; Blotting, Western ; Breast Neoplasms ; enzymology ; genetics ; pathology ; Cell Line, Tumor ; Cell Movement ; drug effects ; Concanavalin A ; pharmacology ; Female ; Humans ; Immunohistochemistry ; Matrix Metalloproteinase 14 ; genetics ; metabolism ; Matrix Metalloproteinase 2 ; metabolism ; Neoplasm Invasiveness ; RNA, Messenger ; genetics ; metabolism

BACKGROUNDMany studies have shown the superior efficacy of budesonide (BUD)/formoterol (FORM) maintenance and reliever therapy, but still lack evidence of its efficacy in Chinese asthma patients in a relative large patient-group. We finished this research to compare BUD/FORM maintenance and reliever therapy and high-dose salmeterol (SALM)/fluticasone (FP) maintenance plus an as-needed short-acting β(2)-agonist in Chinese patients with persistent uncontrolled asthma. This was a post hoc analysis based on a 6-month, multicenter, randomized, double-blind study (NCT00242775).

METHODSA total of 222 eligible asthma patients from nine centers in China were randomized to either BUD/FORM+as-needed BUD/FORM (160/4.5 µg/inhalation) (640/18 µg/d; n = 111), or SALM/FP+as-needed terbutaline (0.4 mg/inhalation) (100/1000 µg/d; n = 111). The primary endpoint was time to first severe exacerbation while secondary endpoints included various measures of pulmonary function, symptom control and quality-of-life.

RESULTSTime to first severe exacerbation over six months was lower with the BUD/FORM than with the SALM/FP treatment (risk ratio = 0.52, 95%CI 0.22 - 1.22), but the difference did not achieve statistical significance (P = 0.13). The cumulative number of severe exacerbations in the BUD/FORM group was lower than in the SALM/FP group (7.2% vs. 13.5%; risk ratio = 0.45, P = 0.028). BUD/FORM produced significantly better improvements in reliever use, cumulative mild exacerbations, symptom-free days (%), and morning/evening peak expiratory flow (PEF) than SALM/FP (P < 0.05 in all cases). The two groups achieved similar improvements in their time to first mild exacerbation, forced expiratory volume in one second (FEV(1)), asthma control questionnaire and asthma symptom scores, and percentage of nights with awakening(s). Both treatments were well tolerated.

CONCLUSIONSIn Chinese patients with persistent asthma, BUD/FORM decreased severe and mild exacerbations, decreased reliever use, increased symptom-free days, and improved morning/evening PEF compared with SALM/FP. There were no significant differences in time to first severe exacerbation or other assessments regarding daily asthma control between BUD/FORM and SALM/FP. BUD/FORM was more effective in this Chinese sub-group than in the total cohort involved in the original study.

Adolescent ; Adult ; Aged ; Asthma ; complications ; drug therapy ; physiopathology ; Budesonide ; administration & dosage ; adverse effects ; Double-Blind Method ; Ethanolamines ; administration & dosage ; adverse effects ; Female ; Forced Expiratory Volume ; Formoterol Fumarate ; Humans ; Male ; Middle Aged

A homology modeling of xylanase XYNB from Streptomyces olivaceoviridis A1 was made by Swiss-Model. The hydrophobic Interaction between beta-sheet B1 and B2 in the tertiary structure model of XYNB was compared with other thermophilic xylanase. A T11Y mutation was introduced in XYNB by site-dirrected mutagenesis to improve the thermostability of the enzyme. The XYNB and mutant xylanase (XYNB') expressed in Pichia pastoris were purified and their enzymatic properties were determined. The result revealed that the thermostability of XYNB' was obviously higher than that of XYNB. The optimal temperature of XYNB' for its activity was 60 degrees C, similar to XYNB. But, compare to XYNB, the optimal pH value, the Km value and the specific activity of XYNB' had also been changed. The research results suggested that the aromatic interaction between beta-sheet B1 and B2 in xylanase should increase enzyme thermostability. The mutant xylanase XYNB' is a good material for further research in the relationship between structure and function of xylanase.
Bacterial Proteins ; chemistry ; genetics ; Endo-1,4-beta Xylanases ; chemistry ; genetics ; Enzyme Stability ; Hot Temperature ; Hydrophobic and Hydrophilic Interactions ; Mutagenesis, Site-Directed ; Mutant Proteins ; chemistry ; Pichia ; genetics ; metabolism ; Protein Conformation ; Protein Folding ; Protein Structure, Tertiary ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Streptomyces ; enzymology ; genetics ; beta-Glucosidase ; chemistry ; genetics

In order to improve the fermentation potency of phytase in recombinant host and decrease the production cost, the pichia expression vector pGAPZalpha-A was modified by introduction of an AOX1 promoter from vector pPIC9 and the resulted vector pAOXZalpha is an methanol induced vector. After that, a phytase gene appA-m was cloned into pAOXZalpha to construct the recombinant vector pAOXZalpha-appA-m. The recombinant Pichia pastoris 74#, which already contains one copy of appA-m and its fermentation potency exceeded 7.5 x 10(6) IU/mL, was used as the host strain for the transformation of pAOXZalpha-appA-m. The Pichia pastoris transformants were gained by electroporation. PCR results indicated that the appA-m expression box has integrated into the genome of Pichia pastoris and the original construction of phytase gene has not changed. SDS-PAGE analysis revealed that phytase was overexpressed and secreted into the medium supernatant. Recombinants with high expression level were screened and used for fermentation. In 5L fermentor, the expression level of phytase protein achieved 4 mg/mL and the phytase activity (fermentation potency) exceeded 1.2 x 10(7) IU/mL, which was about 1.6-fold compared with that of the host strain 74#. Moreover, the improved recombinant Pichia pastoris is excellent at expression stability and heredity stability.
6-Phytase ; genetics ; Fermentation ; Gene Dosage ; Pichia ; genetics ; Plasmids ; Polymerase Chain Reaction ; Recombination, Genetic