AIM: To investigate the influence of Buxinqi Capsule (BXQ) on myocardial ischemia reperfusion injury in rats. METHODS: In this study, the experimental model was established by reperfusion for 60 minutes in rats after ligaturing left anterior descending coronary artery (LAD) for 30 minutes. Serum creatine phosphokinase (CPK), lactate dehydrogenase (LDH), superoxide dismutase (SOD) and malondialdehyodebis (MDA), area of myocardial infarction and occurrence of arrhymia were investigated. RESULTS: BXQ significantly decreased level of CPK and LDH and MDA, and obviously improved the activity of SOD, decreased reperfusion arrhythmias and arrhythmias severity index (ASI), and decreased the area of myocardial infarction. CONCLUSION: BXQ has protective effect on the damage of myocardia ischemia reperfusion in rats

Objective To study the cytotoxicity of cervical collar in rehabilitation technical aids and to investigate the cytotoxicity of domestic marketed products. Methods According to the experimental principle of GB/T 16886.5-2003 and GB/T 16175-2008,microscopic observa-tion and Thiazole Blue Colorimetric methods were used to observe the toxicity of extract from four different cer-vical collars on the cells(L929). Results When the concentration of the extract was 0.1 g/ml,the cytotoxicity was grade three and grade two in the cervi-cal collars D and A,respectively;and it was grade one both in cervical collars B and C.When the concentration of the extract was 0.2 g/ml,the cytotoxicity was grade four in cervical collars A and D,and was grade two in cer-vical collars B and C. Conclusion The cervical collars varied in the cytotoxicity,especially in cervical collars A and D.

OBJECTIVETo establish a mouse model of asthmatic airway remodeling and investigate the effects of 1,25-(OH)2D3 on airway structure and T cell immunoglobulin mucin protein-4 (TIM-4) expression in asthmatic mice.

METHODSThirty female mice (BALB/c strain) were randomly divided into control, asthma and 1,25-(OH)2D3 intervention groups. An asthmatic mouse model was induced using ovalbumin. Lung tissue of the mice was collected, mRNA expression of TIM-4 was evaluated by RT-PCR and airway remodeling and protein expression of TIM-4 were observed by hematoxylineosin staining and immunohistochemistry.

RESULTSTypical airway remodeling was found in the asthma group, and TIM-4 expression in this group was significantly higher than in the control group (105±9 vs 42±5; P<0.05). Compared with the asthma group, the 1,25-(OH)2D3 intervention group showed improvement in airway remodeling and a decrease in TIM-4 expression (78±6) (P<0.05).

CONCLUSIONSTIM-4 may be involved in the airway remodeling of mice. As a new type of immunoregulator, 1,25-(OH)2D3 can downregulate expression of TIM-4 in the lungs and improve airway remodeling in asthmatic mice.

Airway Remodeling ; Animals ; Asthma ; metabolism ; Calcitriol ; pharmacology ; Female ; Gene Expression Regulation ; drug effects ; Lung ; metabolism ; Membrane Proteins ; analysis ; genetics ; physiology ; Mice ; Mice, Inbred BALB C ; RNA, Messenger ; analysis

OBJECTIVETo investigate the effects of 1,25-(OH)(2)D(3) on the airway remodeling and expression of high-mobility group box 1 (HMGB1) and Toll-like receptor 4 (TLR4) in the lungs among asthmatic mice.

METHODSThirty female mice (BALB/c strain) were randomly divided into control, asthma and 1,25-(OH)(2)D(3) intervention groups. An asthmatic mouse model was established by intraperitoneal injection and aerosol inhalation of ovalbumin. The intervention group was given 1,25-(OH)(2)D(3) by intraperitoneal injection 0.5 hour before each aerosol inhalation, while the control group used normal saline instead. The hematoxylin-eosin staining was used to observe the mouse airway structural changes. The mRNA and protein expression of HMGB1 and TLR4 was measured by RT-PCR and immunohistochemistry, respectively. Pearson correlation analysis was performed.

RESULTSThe asthma group had a significantly increased airway wall thickness compared with the control group (P<0.05); the intervention group had a significantly lower increase in airway wall thickness than the asthma group (P<0.05). The mRNA and protein expression of HMGB1 and TLR4 was significantly higher in the asthma group than in the control group (P<0.05); the mRNA and protein expression of HMGB1 and TLR4 in the intervention group was significantly lower than that in the asthma group, but still higher than that in the control group (P<0.05). A positive correlation was found between the protein expression of HMGB1 and TLR4 (P<0.01), and so was their mRNA expression (P<0.01).

CONCLUSIONSHMGB1 and TLR4 may be involved in asthmatic airway remodeling. 1,25-(OH)(2)D(3) can reduce the airway remodeling in asthmatic mice, which may be related to the downregulation of HMGB1 and TLR4 expression in the lungs of asthmatic mice.

Airway Remodeling ; drug effects ; Animals ; Asthma ; drug therapy ; metabolism ; Calcitriol ; pharmacology ; therapeutic use ; Female ; HMGB1 Protein ; genetics ; Lung ; metabolism ; Mice ; Mice, Inbred BALB C ; RNA, Messenger ; analysis ; Toll-Like Receptor 4 ; genetics

OBJECTIVE: To explore the distribution of inflammatory cells and positive expression of P-se- lectin glycoprotein ligand-1 (PSGL-1) in infant brainstem tissue from hand-foot-mouth disease related fatal brainstem encephalitis. METHODS: Twenty brainstem samples from infants suffered from brainstem en- cephalitis were collected as the experimental group. Ten brainstem samples from infants died of non- brain diseases and injuries were collected as the control group. The distribution of inflammatory cells and the expression of PSGL-1 in the two groups were examined by immunohistochemical method. The characteristics of the positive cells were observed. RESULTS: In brainstem tissue of the experimental group, there were sleeve infiltrations of inflammatory cells around the vessels and in the glial nodule. Microglia was the most and following was neutrophils around the vessels and in the glial nodule. There was a significant statistical difference among microglias, neutrophils and lymphocytes (P < 0.05). There was no sleeve infiltration in the control group. PSGL-1 protein was expressed widely in inflammatory cells in the experimental group, especially in the inflammatory cells around the vessels and in the glial nodule. But PSGL-1 positive staining could be observed significantly less in the control group comparing with the experimental group (P < 0.05). CONCLUSION Microglia is the main type of inflammatory cells involved in the progress of the fatal disease. Moreover, PSGL-1 could participate in the pathogenesis of hand-foot-mouth disease related fatal brainstem encephalitis.
Brain Stem/pathology* ; Encephalitis/pathology* ; Hand, Foot and Mouth Disease/pathology* ; Humans ; Infant ; Membrane Glycoproteins/metabolism* ; Microglia/pathology* ; Neutrophils/pathology*

Objective To collect the laboratory parameters and clinical parameters of blood culture positive samples,and analyze the composition and laboratory characteristics of real pathogens by combining with clinical follow-up and anti-infectious treatment outcomes.Methods We conducted a prospective study to isolate the 483 strains of pathogens from 4313 cases of blood samples for clinical examination between March 2013 and March 2015.The results of laboratory and clinical infections were collected for each positive culture and were followed up for clinical follow-up to understand the responsible doctors' experience-based judgment and targeted clinical treatment of antibiotics.After comprehensive analysis we determined the real pathogens and contaminants.Results Of the 483 positive cultures,331 were finally determined as pathogenic ones,accounting for 68.5% of the number of positive isolates; 97 were contaminated bacteria (20.1%); and 55 strains with uncertain pathogenic nature (11.4%).Escherichia coli accounted for the highest proportion (41.2%)of pathogenic bacteria.Coagulase-negative staphylococci took up the highest proportion (75.3%)of the contaminated bacteria.As many as 253 strains (52.4%)were detected from the aerobic or anaerobic bottles.The detection rate of Escherichia coli in anaerobic bottles (23.9%)was higher than that in aerobic bottles (13.8%)(P <0.05).Of 97 strains of positive isolates,only one bottle was reported positive for 90 strains,accounting for (92.8%),and more than two bottles of 7 positive strains,accounting for (7.2%)(P <0.05).34 positive in 24 h (35.1%),77 positive in 48 h (79.4%),the positivebacteria ratio within 48 h (79.4%)was higher than that of bacteria contamination ratio within 24 h (χ2 =38.935, P =0.000),with a significant difference.Conclusion Establishment of contaminated bacteria in blood culture cannot rely solely on laboratory or clinical parameters.It should be combined with the experience of clinicians to determine the clinical response to comprehensive judgments.For the laboratory to determine the presence of contamination,the number of positive bottles and the amount of sun are still two factors of important value.Paying attention to inspection of anaerobic bottles is more conducive to the detection of Escherichia coli.

OBJECTIVETo determine whether there are any associations between the -258T/G polymorphism of the promoter and the IVS3 -20T/C polymorphism in parkin gene and Parkinson's disease (PD) from a Han population in Sichuan province.

METHODSPolymerase chain reaction (PCR), restriction fragment length polymorphism, denaturing high performance liquid chromatography(dHPLC) and sequence analysis were used to determine the genotype of each subject. The -258T/G polymorphism and IVS3 -20T/C polymorphism were analysed in 198 patients with sporadic PD and 187 healthy controls, matched for age and gender.

RESULTSThere were significant differences in allele frequency of the -258T/G polymorphism between PD patients and controls, with the G allele more common in cases than controls (52.5% vs 43.3%; chi square is 6.17, P< 0.025, OR is 1.45, 95% CI 1.04-1.86). There were also significant differences in G allele frequency between PD patients with onset age over 50 years old and controls(chi square is 9.048, P< 0.01, OR is 1.57, 95% CI:1.08-2.06). The frequency of TG+GG genotype was significantly higher in PD patients than in controls (78.79% vs 69.51%; chi square is 3.854, P< 0.05, OR is 1.63, 95% CI:0.88-2.38). In addition, there were significant differences in age of onset between PD patients with different genotypes (P< 0.05). The average age of onset in group of GG genotype was later about 5 years compared with the group of TT or TG genotype. The frequency of CC genotype in IVS3 -20T/C polymorphism was much higher than that of TC genotype. No TT genotype was found.

CONCLUSIONThis study suggests that the parkin promoter -258T/G polymorphism might be a risk factor for late onset PD in Sichuan. CC genotype for IVS3 -20T/C polymorphism is common in Sichuan Han population. No TT genotype for IVS3 -20T/C polymorphism is found in Sichuan Han population.

Adult ; Aged ; Aged, 80 and over ; Asian Continental Ancestry Group ; genetics ; China ; Chromatography, High Pressure Liquid ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; genetics ; Genotype ; Humans ; Male ; Middle Aged ; Parkinson Disease ; ethnology ; genetics ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Polymorphism, Restriction Fragment Length ; Promoter Regions, Genetic ; genetics ; Ubiquitin-Protein Ligases ; genetics

OBJECTIVETo compare and evaluate the clinical value among transperitoneal, retroperitoneal laparoscopic adrenalectomy and open adrenalectomy for the treatment of adrenal tumours.

METHODSFrom January 1996 to June 2002, transperitoneal laparoscopic adrenalectomy (TLA) was performed for 19 cases, 17 cases were successful (Group A), retroperitoneal laparoscopic adrenalectomy (RLA) for 21 cases, 19 cases were successful (Group B), during the same period 41 patients with adrenal tumours under 6 cm in diameter underwent open adrenalectomy (Group C). The procedure was evaluated and compared in the respects of tumors weight, operation time, blood loss, recovery time of GI function, hospital stay, complications transferable rate and indications among three groups.

RESULTSAverage tumors weight was (13.86) g in group A, (15.66) g in group B and (18.03) g in group C; mean operating time: group A was 17.21 min longer than group B, group A. B was 48.53 min longer and 31.32 min longer han group C respectively, mean blood loss in group A. B was less diminished by 121.55 ml and 137.05 ml than group C, and the blood loss in group B was less diminished by 15.50 ml than group A; mean recovery of Gl function, group A and B was 1.87 day and 2.17 day earlier than group C respectively, group B was 0.3 day earlier than group A, mean hospital stay was 2.60 day and 3.87 day shorter than group C, group B was 1.27 day shorter, than group A; transferable rate of operation was 10.5% (group A) and 9.5% (group B). Postoperative complication rate was 10.5%, 14.3% and 9.7% in group A. B and C respectively.

CONCLUSIONSTLA and RLA were better than open method. On the hand of approach to adrenal gland disturbs to abdominal organ and postoperative recovery on RLA have some slight advantage.

Adrenal Gland Neoplasms ; surgery ; Adrenalectomy ; methods ; Adult ; Aged ; Female ; Humans ; Laparoscopy ; methods ; Laparotomy ; Male ; Middle Aged ; Postoperative Complications ; prevention & control ; Retroperitoneal Space ; Retrospective Studies ; Treatment Outcome

Aspergillus fumigatus wild-type phytase has many favorable properties, such as a good thermorstability and a broad pH optimum. However, the specific activity of the enzyme is relative low. A. fumigatus Q23L phytase resulted in a remarkable increase in specific activity around pH4.5 - 7.0, but the pH stability of Q23L was lower than A. fumigatus wild-type phytase. To increase the pH stability of Q23L, the mutant Q23LG272E was constructed by site-directed mutagenesis with PCR. The gene of A. fumigatus wild-type phytase and the mutant genes encoding the Q23LG272E and the Q23L were correctly expressed in Pichia pastoris GS115. Enzymes were purified and their enzymatic properties were determined. The results revealed that the specific activity of the Q23L improved remarkably, which increased from 51 u/mg of the wild type to 109 u/mg at pH5.5. Meanwhile, the pH stability of Q23L, decreased evidently, especially from pH3.0 to pH4.0.The pH stability of Q23LG272E in pH3.0 - 4.5 and pH6.5 - 7.0 has been improved compared with Q23L. The specific activity of Q23LG272E basically maintained at the level of Q23L. Analysis of 3-D structure and sequence similarity were used to reveal the presumable factors influencing the enzymatic properties of Q23LG272E, and discussion for the relationship between structure and function of phytase was given.
6-Phytase ; chemistry ; genetics ; metabolism ; Amino Acid Substitution ; Aspergillus fumigatus ; enzymology ; genetics ; Biocatalysis ; Electrophoresis, Polyacrylamide Gel ; Fungal Proteins ; chemistry ; genetics ; metabolism ; Hydrogen-Ion Concentration ; Models, Molecular ; Mutagenesis, Site-Directed ; Mutant Proteins ; genetics ; metabolism ; Mutation ; Pichia ; genetics ; Polymerase Chain Reaction ; Protein Conformation ; Protein Engineering ; methods ; Recombinant Proteins ; metabolism ; Structure-Activity Relationship ; Substrate Specificity

A gene appA encoding a novel phytase was firstly cloned from Hafnia alvei by PCR and sequenced. The gene was consisted of 1335 bp, encoding 444 amino acids. The calculated molecular weight of the mature APPA was about 45.2 kD. The gene appA was expressed in E. coli BL21 (DE3). Recombinant APPA was purified and its enzymatic properties were determined. The optimum pH for the enzyme was 4.5 and the optimum temperature was 60 degrees C. The pH stability of r-APPA is good, the relative phytase activity was above 80% after treated in buffers of pH 2.0-10.0. The specific activity of r-APPA is 356.7 U/mg, and the Km value was 0.49 mmol/L and Vmax of 238 U/mg. The enzyme showed resistance to pepsin and trypsin treatment.
6-Phytase ; biosynthesis ; genetics ; isolation & purification ; Amino Acid Sequence ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Hafnia ; enzymology ; genetics ; Hydrogen-Ion Concentration ; Molecular Sequence Data ; Recombinant Proteins ; biosynthesis ; genetics ; isolation & purification ; metabolism ; Temperature