Objective To establish a new determination method for the measuring of alkaline phosphatase activity (ALP) with p-acetyl phenyl phosphace (PAP-PNa_2) as substrate.Methods With the help of Vital semiautomatic analyzer,researched a continuous-monitoring procedure and set up experimental parameters.Results When using this assay,the wavelength of PAP's absorption was 325 nm and the Km of ALP was 0.376 mmol/L.The molecular extinction coefficient of PAP at 340 nm was 23 390 L?mol~(-1)? cm~(-1) and the concentration of citrate buffer was 0.438 mol/L.During the process,we found that the optimum pH of enzyme was 10.4,and the concentration of substrate was 5.0 mmol/L.The time of linear reaction was 900 seconds,and the linear range was 0-1 110 U/L.Serum total ALP were 63.1-118.3 U/ L(male) and 52.5-89.0 U/L(female),based on results from 60 heath adults.Conclusions The method is practical in its repetition and convenience,saves time and is not liable to be affected by bilirubin in serum.It is especially suited to the use of automatic analyzers.

Carcinoma ; genetics ; metabolism ; pathology ; surgery ; China ; Humans ; Male ; Oncogene Proteins, Fusion ; genetics ; Prostatic Hyperplasia ; genetics ; metabolism ; pathology ; surgery ; Prostatic Neoplasms ; genetics ; metabolism ; pathology ; surgery ; Proto-Oncogene Proteins c-ets ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Serine Endopeptidases ; genetics ; metabolism

Adult ; Disease Outbreaks ; Humans ; Metapneumovirus ; Paramyxoviridae Infections ; epidemiology ; Respiratory Tract Infections ; epidemiology ; virology ; Students

OBJECTIVETo explore if reduced number of circulating endothelial progenitor cells (EPCs) is a risk factor for patients with coronary slow flow (CSF).

METHODSThirty patients with CSF and 30 age and gender matched control subjects with normal coronary angiography were included in the study. Mononuclear cells were isolated from peripheral blood by Ficoll density gradient centrifugation and plated on fibronectin-coated culture dishes. EPCs were characterized as adherent cells double positive for DiI-AcLDL-uptake and lectin-binding by converted fluorescence microscope (×200).

RESULTSSmoking, diabetes mellitus, hypertension and the levels of plasma lipoprotein profile were similar between the two groups (all P > 0.05). The number of EPCs was significantly lower in patients with CSF compared with control subjects (35.7 ± 5.9 vs.53.2 ± 5.9, P < 0.01). TIMI frame counts was correlated with circulating EPCs number (OR = 0.424, 95%CI 0.358 - 0.621, P < 0.01) and not associated with gender, age, smoking, diabetes mellitus, hypertension and the levels of plasma lipoprotein profile.

CONCLUSIONDecreased circulating EPCs is an independent risk factor for CSF.

Blood Circulation ; Blood Flow Velocity ; Case-Control Studies ; Cell Count ; Cells, Cultured ; Coronary Angiography ; Coronary Vessels ; physiopathology ; Female ; Humans ; Male ; Middle Aged ; Risk Factors ; Stem Cells ; cytology

Objective: To evaluate the efficiency and safety of low intensity conditional regimen for children with Fanconi anemia (FA) receiving allogenic hematopoietic stem cells transplantation (allo-HSCT). Methods: Four patients diagnosed as Fanconi anemia were enrolled in this study. One patient received HLA-identical sibling donor hematopoietic stem cell transplantation, two patients underwent unrelated donor matched (UD) HSCT, and one patient received unrelated cord blood transplantation. The conditional regimen consisted of Busulfan with low dose of cyclophosphamide. Results: All 4 cases succeeded in allo-HSCT. The median time for neutrophils engraftment was 11(9-15) day, median time to platelets (PLT) engraftment was 12 (8-28) day. One case occurred with grade I of aGVHD, 1 case with hemorrhagic cystitis. No patient happened with hepatic veno-occlusive disease (VOD). Conclusion: Low intensity of conditional regimen is efficient and safe which should be recommended for FA patients with HSCT.
Busulfan ; Fanconi Anemia ; Graft vs Host Disease ; Hematopoietic Stem Cell Transplantation ; Humans ; Transplantation Conditioning

Objective To study the effect of dynamic pressure on the expression of parathyroid hormone-related protein (PTHrP)mRNA in metaphyseal cartilage stem cells of rats so as to further explore whether fiber actin (F-actin)is involved in the mechanical signal transduction process.Methods We isolated and cultured metaphyseal cartilage stem cells of rats by immunomagnetic beads.The third-generation rat metaphyseal cartilage stem cells were randomly divided into four groups:0%,3%,6%,and 12% deformed groups according to the size of dynamic pressure strength.We used a self-prepared dynamic tonic culture device to exert different intensity of pressure on each group of cells for 24 hours.Flow cytometry was used to detect the cell cycle distribution and apoptosis rate.The expression of PTHrP mRNA in each group was detected by Rea-l time quantitative PCR. Furthermore,the third-generation rat metaphyseal cartilage stem cells were randomly divided into four groups:control group,simple pressure group (6% deformation),pressure+cytoskeleton relaxin D group,and simple cytoskeleton relaxin D group according to whether or not to apply pressure and cytoskeleton relaxin D.F-actin fibers in each group of cells were stained with phalloidin and placed under a laser scanning confocal microscope.The expression of PTHrP mRNA in each group was detected by Real-time quantitative PCR.Results The results of flow cytometry showed no significant difference in G0/G1,G2/M and S phases between 0%,3%,6% and 12% deformed groups (P＞0.05).There was no significant difference in the apoptosis rate between 3% and 6% deformed groups compared with 0% deformed group (P＞0.05).The apoptosis rate was significantly higher in 1 2 % deformed group than in control group (P＜0.05).The results of laser confocal microscopy showed that the arrangement of F-actin fibers in the pressure group was neat and parallel compared with that in the control group, which was consistent with the direction of force.The intracellular F-actin fiber structure in pressure+cytoskeleton relaxin D group and simple cytoskeleton relaxin D group was destroyed and aggregated into clusters.Real-time quantitative PCR results showed that PTHrP mRNA expression did not significantly differ between 3% and 0% deformed groups (P＞0.05).The expression of PTHrP mRNA in 6% and 12% deformed groups was significantly higher than that in 0% group (P＜0.05).The expression of PTHrP mRNA in the cells of simple pressure group was significantly higher than that in the control group (P＜0.05).There was no significant difference in the expression of PTHrP mRNA between simple cytoskeleton relaxin D group and control group (P＞0.05).The mRNA expression of PTHrP was higher in pressure+cytoskeleton relaxin D group than that in control group,but lower than in simple pressure group (P＜0.05).Conclusion The dynamic pressure of proper intensity can increase the mRNA expression of PTHrP in chondrocytes of metaphyseal hypertrophy in rats,and F-actin is involved in the mechanical signal transduction process.

Objective:To investigate the clinical significance of preserving autonomic nerve (PANP) in patients with low rectal cancer.Methods:The clinical data of 68 cases of patients with low rectal cancer who underwent laparoscopic surgery in our hospital from March 2014 to January 2016 were retrospectively analyzed.According to the surgical methods,the patients were divided into PANP group and non PANP group.Two groups of patients with postoperative micturition function,sexual function and quality of life were observed,compared with the two groups of patients with postoperative satisfaction differences.Results:In retention of PANP group of patients with postoperative ultrasonic residual urine volume was lower than that in non retention in PANP group were less and recovery of automatic micturition time is short retention in PANP group,IPSS score was lower than that in non low retention in PANP group,the difference is statistically significant(P＜0.05);The female and male patients with retention of PANP group were better than those of non retention PANP group,and the difference was statistically significant (P＜0.05);There was no significant difference in quality of life between the two groups before surgery.After surgery,the quality of life of the two groups was lower than before treatment,and the decrease of PANP group was more obvious than that of the non retention group (P＜0.05).Conclusions:The autonomic nerve preservation in the operation of low rectal cancer has a good protective effect on the urinary function and sexual function,and can obviously improve the quality of life of patients after operation.

OBJECTIVETo study the relationship between plant growth and accumulation of active ingredients in Salvia miltiorrhiza.

METHODTransplants of S. miltiorrhiza were sampled at 20 day intervals. At each stage, the growth of seedling and root system was recorded and the contents of tanshinone II (A) and salvia acid were measured.

RESULT AND CONCLUSIONThis study showed that the rapid growth stage of the root system lags behind that of the seedling system, but the growing period of root system lasts longer. The quantitative change of roots reveals a double "S" curve; two rapid growth stages emerge during 30 - 70 days and 140 - 200 days after the seedlings were transplanted. The content of salvia acid reaches the highest level during 140 - 180 days, whereas the content of tanshinone peaks during 100 - 120 days.

Agriculture ; methods ; Diterpenes, Abietane ; Phenanthrenes ; analysis ; Plant Roots ; chemistry ; growth & development ; Plants, Medicinal ; chemistry ; growth & development ; Salvia miltiorrhiza ; chemistry ; growth & development ; Seedlings ; chemistry ; growth & development

OBJECTIVETo explore the effects of Dangua Recipe (DGR) on glycolipid metabolism, serum reactive oxygen species (ROS) level, nuclear factor kappa B (NF-kappaB) positive expression and its mRNA expression level in the thoracic aorta of diabetic rats with atherosclerosis, thus revealing its partial mechanisms for intervening chronic diabetic complications.

METHODSRecruited 40 Goto-Kakisaki (GK) Wistar rats were fed with high fat forage containing metabolic inhibition Propylthiouracil, and peritoneally injected with endothelial NOS inhibitor N-nitro-L-arginine methyl ester to establish a high fat diabetes model with atherosclerosis. The modeled GK rats were stratified by body weight, and then, by blood glucose level from high to low, randomly divided into the DGR group (at the daily dose of 8 mL/kg), the metformin group (MET, at the daily dose of 150 mg/kg), the simvastatin group (SIM, at the daily dose of 2 mg/kg), and the model group (MOD, fed with pure water, at the daily dose of 8 mL/kg) according to the random number table, 10 in each group. Another 10 Wistar rats of the same ages and comparable body weight level were recruited as the normal control group. All the interventions lasted for 24 weeks by gastrogavage. The fasting blood glucose (FBG) and body weight were monitored. The HbA1c, TC, LDL-C, HDL-C, TG, serum ROS were determined. The aortic NF-kappaB level was analyzed with immunohistochemical assay. The expression of NF-kappaB (P65) mRNA in the aorta was detected with Real-time PCR.

RESULTSThe body weight in the normal control group was eventually heavier than others (P < 0.01). There was no difference among the four groups of GK modeled rats (P > 0.05). The FBG in the four GK modeled groups were higher than that in the normal control group (P < 0.01, P < 0.05). There was no statistical difference in the blood glucose level at the first visit and at the baseline among the GK modeled groups (P > 0.05). The last FBG level was obviously lower in the MET and DGR groups than in the MOD group (P < 0.01) and the SIM group (P < 0.05). Twenty-four weeks after intervention, the level of FBG, HbA1c, TC, LDL-C, HDL-C, and NF-kappaB positive expression rate of the thoracic aorta of the four groups of GK modeled rats, and NF-kappaB mRNA expression in the thoracic aorta in the MOD group, the MET group, and the DGR group were significantly higher than those in the normal control group (P < 0.01, P < 0.05). The TG level, serum ROS in the MET, DGR, and SIM groups, and the NF-kappaB mRNA expression level in the thoracic aorta in the SIM group were significantly lower than those in the normal control group (P < 0.01, P < 0.05). The levels of FBG, TC, LDL-C, serum ROS, NF-kappaB mRNA expression level in the thoracic aorta in three drug intervention groups, and NF-kappaB positive expression rate in the DGR and MET groups, and the levels of HbA1c, TG in the DGR group were significantly lower than those in the MOD group (P < 0.01, P < 0.05). The level of FBG in the MET and DGR groups were lower than that in the SIM group (P < 0.05). The level of NF-kappaB mRNA expression in the thoracic aorta of the SIM and DGR groups, and the levels of TC and LDL-C in the DGR group were significantly lower than those in the MET group (P < 0.01).

CONCLUSIONDGR played a role in preventing and treating chronic diabetic complications by comprehensively regulating blood glucose and serum lipids, as well as down-regulating oxidative stress.

Animals ; Aorta, Thoracic ; metabolism ; Atherosclerosis ; complications ; drug therapy ; metabolism ; Blood Glucose ; analysis ; Diabetic Angiopathies ; drug therapy ; metabolism ; Disease Models, Animal ; Drugs, Chinese Herbal ; therapeutic use ; Lipid Metabolism ; Male ; NF-kappa B ; metabolism ; Oxidative Stress ; Phytotherapy ; Rats ; Rats, Wistar ; Reactive Oxygen Species ; blood

OBJECTIVETo characterize the profile of chromosomal imbalances in esophageal cancer (EC) with or without family history in Linzhou, Henan Province of China.

METHODSComparative genomic hybridization (CGH) was used to examine 13 cases with positive family history of EC and 32 cases with negative family history of EC. RESULTS DNA copy number gains on chromosome 10q was observed only in the cases with postivie family history of EC (30%), and none in cases with a negative family history (P<0.05). DNA copy number losses on chromosome 15q were significantly higher in cases with postivie family history (38% vs 6%, P<0.05). The frequency of DNA copy number gains in 3q, 5p, 7p, 8q and DNA copy number losses in 3p, 19q, 9q were similar in the two groups (both beyond 20%) (P>0.05).

CONCLUSIONSFrequent DNA copy number gains on chromosome 10q and losses on chromosome 15q in EC casers with postivie family history indicate that these chromosome sites may harbor the genes related to high susceptibility to EC. Such chromosomal sites as 3q, 5p, 7p, 8q, 3p, 19q, and 9q may contain important genes related with the environmental risk factors of esophageal carcinogenesis.

Adult ; Aged ; China ; Chromosome Disorders ; genetics ; Chromosomes, Human, Pair 10 ; genetics ; Chromosomes, Human, Pair 15 ; genetics ; Comparative Genomic Hybridization ; methods ; Esophageal Neoplasms ; genetics ; Family Health ; Female ; Gene Deletion ; Genetic Predisposition to Disease ; genetics ; Humans ; Male ; Middle Aged