Objective:To determine the valproate concentration in plasma of epilepsy patients by HPLC, and compare with the re-sults of chemiluminescence microparticle immunoassay ( CMIA) to evaluate the consistency of the two methods. Methods:HPLC and CMIA was respectively applied to determine the plasma concentration of valproate in 230 epileptic patients. The correlation of the two methods was studied by Passing-Bablok regression and Bland-Altman method. Results:The regression equation of the determination re-sults of HPLC (Y) and CMIA (X) was Y=1. 069 7X+2. 338 2 (R2 =0. 969, n=230), which showed promising correlation. Bland-Altman analysis showed that the consistency of the two methods was poor, and the values of HPLC were higher. Conclusion: HPLC and CMIA used for the determination of valproate plasma concentration show good correlation. However, the consistency is poor and there is system error. In the clinical treatment, adjustment and choice should be paid more attention.

Objective To establish a high performance liquid chromatography (HPLC) method for determining phenytoin concentration in epilepsy patients' plasma,and compare this method with chemiluminescence microparticle immunoassay (CMIA),and to evaluate the consistency of the two methods.Methods HPLC and CMIA methods were applied to determine the plasma concentration of phenytoin in 60 epileptic patients,respectively.The difference of results was analyzed by two-side paired t-test,and then the correlation and consistency of the two methods were investigated with Passing-Bablok regression and Bland-Altman method.Results There was no significant difference between the results of the two methods (P ＞0.05).The regression equation of the determination results by HPLC (Y) and CMIA (X) was Y=0.992 9X +0.143 7 (R2 =0.992 6,n =60),which indicated the correlation of the two methods was good.Bland-Altman analysis showed that the consistency of the two methods for determining was good.Conclusion HPLC and CMIA method in monitoring plasma concentration of phenytoin have good correlation and consistency.Both methods can be used for therapeutic drug monitoring of phenytoin.

Abdominal Pain/etiology* ; Acute Disease ; Appendicitis/diagnosis* ; Diabetes Mellitus ; Diabetic Ketoacidosis/diagnosis* ; Diagnostic Errors/adverse effects* ; Humans ; Insulin

This study was purposed to investigate the effects of tortois plastron, astragali, salviae miltiorrhizae and codonopsis pilosulae on gamma-globin gene synthesis in K562 cells in vitro. Benzidine staining was used to clarify the dose-and time-dependent effects of tortois plastron, astragali, salviae miltiorrhizae and codonopsis pilosulae on hemoglobin synthesis in K562 cells and Western blotting was performed to determine the level of hemoglobin F (alpha(2)gamma(2)). The results indicated that the K562 cells treated with 4 kinds of traditional Chinese medicine had different stain rates of benzidine for: 23.5% (tortois plastron), 19.8% (astragali), 15.8% (salviae miltiorrhizae) and 14.5% (codonopsis pilosulae) at 6 days after the treatment. Western blot indicated that synthesis of HbF increased. It is concluded that tortois plastron, astragali, salviae miltiorrhizae and codonopsis pilosulae enhance globins-gamma synthesis level and increase hemoglobin F level in K562 cells, the effect of which resembles that of sodium butyrate.
Animals ; Astragalus membranaceus ; chemistry ; Codonopsis ; chemistry ; Drugs, Chinese Herbal ; pharmacology ; Female ; Humans ; K562 Cells ; Male ; Materia Medica ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Salvia miltiorrhiza ; chemistry ; gamma-Globins ; biosynthesis

OBJECTIVETo investigate the absorption mechanism of genistein self-microemulsifying system in rat intestines.

METHODThe concentrations of phenol red and genistein by in situ perfusion in rats were determined by UV and HPLC, respectively. The effects of drug concentrations, pH, various intestinal segments and P-glycoprotein (P-gp) inhibitor verapamil on the absorption had been studied.

RESULTThe absorption rate constant (Ka) of genistein had no significant difference at concentrations of 0.05-0.5 mg x mL(-1) and pH of 5.4-7.8 in perfusion. It was Ka of jejunum > ileum > duodenum > colon. The absorption of genistein in jejunum had significant difference (P < 0.05) compared with other parts of intestines. Ka was increased obviously when verapamil was coper-fused with genistein (P < 0. 05).

CONCLUSIONThe absorption of genistein self-microemulsifying system is a first order process with passive diffusion mechanism related to P-gp efflux. It can be absorbed at all segments of rat intestine, and the jejunum is the best absorption segment, pH had no special effect on the absorption of genistein self-microemulsifying system in rat intestine.

ATP Binding Cassette Transporter, Sub-Family B ; antagonists & inhibitors ; Animals ; Chromatography, High Pressure Liquid ; Emulsions ; Genistein ; analysis ; metabolism ; pharmacokinetics ; Hydrogen-Ion Concentration ; Intestinal Absorption ; drug effects ; Intestines ; drug effects ; metabolism ; Male ; Organ Specificity ; Rats ; Rats, Wistar ; Temperature ; Verapamil ; pharmacology

The present study was aimed to investigate the effects of hypoxia on iron metabolism of skeletal muscle. Rat L6 skeletal muscle cells were randomly divided into three groups which were exposed to hypoxia (1% O(2)) for 0, 12, 24 h, respectively. Iron isotope tracing method was used to determine iron uptake and release. Iron content of labile iron pool (LIP) was investigated by flow cytometry, and the expressions of transferrin receptor 1 (TfR1), divalent metal transporter 1 (DMT1), ferroportin 1 (FPN1), hypoxia-inducible transcription factor-1 (HIF-1) in L6 cells were observed by Western blot. The results showed that, compared with 0-hour hypoxia group, 12-hour hypoxia group exhibited significantly increased iron uptake and LIP (P < 0.05), as well as decreased iron release (P < 0.01). Not only iron uptake and release, but also LIP in 24-hour hypoxia group were significantly decreased, compared with those in 0- and 12-hour hypoxia groups (P < 0.01). The expressions of HIF-1, DMT1 (IRE), DMT1 (non-IRE) and TfR1 in 12-hour hypoxia group were significantly increased compared with those in 0-hour hypoxia group (P < 0.01). On the contrary, the expressions of DMT1 (IRE), DMT1 (non-IRE) and FPN1 in 24-hour hypoxia group were significantly decreased compared with those in the other two groups. However, TfR1 expression in 24-hour hypoxia was higher than those in 0- and 12-hour hypoxia groups (P < 0.05 or P < 0.01). These results suggest hypoxia plays an important role in iron metabolism of skeletal muscle cells. Moderate hypoxia can increase iron uptake and decrease iron release, resulting in higher LIP, but a prolonged hypoxia induces a disordered iron metabolism of skeletal cells.
Animals ; Cation Transport Proteins ; metabolism ; Cell Hypoxia ; Cell Line ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Iron ; metabolism ; Muscle, Skeletal ; cytology ; metabolism ; Oxygen ; physiology ; Rats ; Receptors, Transferrin ; metabolism ; Time Factors

OBJECTIVETo analyze the differential proteomics in human umbilical cord mesenchymal stem cells (MSC) induced by chemical hypoxia-mimetic agent cobalt chloride (CoCl(2)) by two-dimensional gel electrophoresis (2-DE) and mass-spectrometry.

METHODS2-DE was performed to separate proteins from treated and untreated human umbilical cord MSC with CoCl(2). 2-DE images were analyzed by ImageMaster 2D Platinum software 6.0. The differential expressed proteins was identified by MALDI-TOF-MS. The differential proteins were classified based on their functions.

RESULTS2-DE reference patterns of CoCl(2) treated human umbilical cord MSC were established. A total of twenty-six differential proteins were identified, of them eleven proteins were up-regulated and fifteen down-regulated. Their biological functions involved in carbohydrate metabolism, protein metabolism and modification, lipid metabolism, coenzyme and prosthetic group metabolism, cell cycle, immunity and defense, cell structure and motility, signal transduction, protein targeting and localization, neuronal activities, muscle contraction, etc. Peroxiredoxin1 (Prdx) was down-regulated, whereas alpha-enolase (ENO1) and vesicle amine transport protein 1 homolog (VAT1) up-regulated.

CONCLUSIONThe effects of hypoxia on human umbilical cord MSC were participated by multiple proteins and involved in multiple functional pathways.

Cobalt ; pharmacology ; Humans ; Mesenchymal Stromal Cells ; drug effects ; metabolism ; Proteome ; analysis ; Proteomics ; Umbilical Cord ; cytology ; drug effects

OBJECTIVEIn order to investigate whether the presence of distant metastases is associated with serum lipid abnormalities.

METHODSThe fasting serum lipid profile and various clinicopathological data of 324 breast cancer patients with and without synchronous distant metastases were collected and analyzed. The serum lipid profile, including total cholesterol (TC), triglycerides (TG), low-density (LDL-C) and high-density lipoprotein cholesterol (HDL-C) was determined. The nutritional status, the serum albumin was measured and body mass index (BMI) was calculated. Univariate analysis and multiple logistic regression analysis were carried out to investigate the association of serum lipid profile with distant metastases.

RESULTSUnivariate analysis showed that the distant metastasis rate was significantly higher in the breast cancer patients with an higher level of serum TC, TG, LDL-C, and LDL-C/HDL-C ratio (P < 0.05). Multiple logistic regression analysis showed that higher serum levels of TC, LDL-C and LDL-C/HDL-C ratio were independent risk factors for distant metastasis in breast cancer (OR = 2.324, 2.648 and 4.862, respectively).

CONCLUSIONSHyperlipidemia is significantly associated with the distant metastasis in breast cancer patients. Monitoring of serum lipid profile may be helpful to predict the occurrence of distant metastasis in breast cancer patients.

Adult ; Aged ; Aged, 80 and over ; Body Mass Index ; Breast Neoplasms ; blood ; pathology ; Cholesterol ; blood ; Cholesterol, HDL ; blood ; Cholesterol, LDL ; blood ; Female ; Humans ; Lipids ; blood ; Middle Aged ; Multivariate Analysis ; Neoplasm Metastasis ; Neoplasm Staging ; Nutritional Status ; Risk Factors ; Serum Albumin ; Triglycerides ; blood

OBJECTIVETo investigate the role of duodenum in regulation of ghrelin and body mass index (BMI) and the correlation between ghrelin and BMI after subtotal gastrectomy.

METHODSForty-two patients with T(0-1)N(0-1)M(0) gastric cancer were divided into two groups after gastrectomy according to digestive reconstruction pattern, Billroth I group (n=23) and Billroth II group (n=19) respectively. Plasma ghrelin levels were determined by radioimmunoassay (RIA) before and at day 1, 7, 30 and 360 after gastrectomy,and BMIs were also measured.

RESULTSTwo groups had identical postoperative trends in ghrelin alterations during the early stage, both dropping to nadir at day 1 (36.7% vs 35.7%), then markedly increasing at day 7 (51.0% vs 51.1%). At day 30, ghrelin level of Billroth I group was slightly higher than that of Billroth II group. At day 360, ghrelin level of Billroth I group recovered to 93.6%, approaching though lower than preoperative level and no significant difference was displayed, while ghrelin level of Billroth II group recovered only to 81.6% of preoperational level and significant difference existed (P=0.033). Compared with preoperative levels, ghrelin of two groups decreased by 6.9% and 18.4% while BMI by 3.3% and 6.4% respectively, liner regression correlations were revealed in both groups between decrease magnitudes(R(1)(2)=0.297,P=0.00;R(2)(2)=0.559,P<0.001).

CONCLUSIONSAnatomico-physiological duodenum compensatively promotes ghrelin recovery, accordingly enhances BMI after gastrectomy. Regarding patients with insufficient ghrelin secretion, ghrelin is positively correlated with BMI.

Adult ; Body Mass Index ; Duodenum ; metabolism ; Female ; Gastrectomy ; methods ; Ghrelin ; blood ; Humans ; Male ; Middle Aged ; Stomach Neoplasms ; blood ; physiopathology ; surgery

OBJECTIVETo clone the gene human thioredoxin 1 (hTrx-1) expressing its protein in the E.coli expression system and to obtain its polyclonal antibody, and to study the protective effects of hTrx-1 on neonatal rats with endotoxemia.

METHODSDNA encoding hTrx-1 from fetal liver cells was isolated by RT-PCR. The hTrx-1 was cloned to the prokaryotic expression plasmid PET-28a to induce its protein expression in the E.coli expression system. The purified hTrx-1 was injected into rats to prepare polyclonal antibody. Newborn Sprague-Dawley rats were randomly assigned to three groups: control, lipopolysaccharide (LPS) and hTrx-1 (n=12 each). The control and the LPS groups were intraperitoneally injected with normal saline and LPS (5 mg/kg), respectively. The hTrx-1 group received an intraperitoneal injection of hTrx-1 (10 mg/kg) 30 minutes before LPS injection. The mortality rate 24 hrs after injection was compared between the three groups.

RESULTSThe prokaryotic expression plasmid PET-28a-hTrx-1 was constructed. The hTrx-1 protein was expressed and purified. The polyclonal antibody of hTrx-1 with the titer of 1∶51200 was prepared. The mortality rate of the control, LPS and hTrx-1 groups was 0, 67% and 17%, respectively (χ2=14.400, P<0.01).

CONCLUSIONSThe polyclonal antibody of hTrx-1 is prepared successfully. The hTrx-1 protein has protective effects on neonatal rats with endotoxiamia.

Animals ; Animals, Newborn ; Antibodies ; analysis ; Blotting, Western ; Endotoxemia ; prevention & control ; Enzyme-Linked Immunosorbent Assay ; Humans ; Male ; Rats ; Rats, Sprague-Dawley ; Recombinant Proteins ; immunology ; Thioredoxins ; genetics ; immunology ; therapeutic use