[ABSTRACT]AIM:Toinvestigatetheroleofcysteine-rich61(Cyr61/CNN1)inproliferationandmigrationof bone marrow mesenchymal stem cells ( BMSCs ) .METHODS: The lentiviral vector carrying CCN 1 ( Lenti-GFP-CCN1 ) was constructed and then transfected into the rat BMSCs .The cells were divided into non-transfection group , transfection group ( transfected with Lenti-GFP-CCN1 ) and negative control group ( Lenti-GFP ) .The fluorescence intensity of the transfected BMSCs was observed under inverted fluorescence microscope .The effects of CCN1 on the proliferation and mi-gration of BMSCs were detected by MTT assay and scratch wound healing assay .RESULTS:The proliferation of BMSCs transfected with Lenti-GFP CCN1 had no significant difference compared with negative control group and control group .The width/thickness ratio of migrated BMSCs in wound healing was significantly higher in Lenti-GFP-CCN1 group than that in negative control group and control group (P<0.05).CONCLUSION:Exogenous CCN1 promotes the migration of BMSCs.

As a keratinized material, nail recently has attracting researchers' attention in the pharmaceuticals analysis. There are comparatively limited studies concerning nail's xenobiotic determination and its mechanism. This article reported the development of a sensitive, specific and reproducible LC-MS/MS method, which could be as a foundation of other studies on drug determination in nail. It can also be regarded as the first report on organic drug in mainland China. Sixteen nail samples from volunteers, who were ingested clozapine for more than nine months, are confirmed positive after being analyzed by the method. It is found that contents of clozapine in the patients' nails are above the nanogram level. Besides, a comparative study of clozapine concentration in nails and hair was made, with a result that there exists a correlation between the two materials in terms of clozapine concentration.
Adult ; Antipsychotic Agents ; pharmacokinetics ; China ; Chromatography, Liquid ; methods ; Clozapine ; pharmacokinetics ; Female ; Hair ; chemistry ; Humans ; Male ; Middle Aged ; Nails ; chemistry ; Psychotic Disorders ; metabolism ; Tandem Mass Spectrometry ; methods

OBJECTIVE: To establish a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the simultaneous analysis of cocaine (COC) and its metabolite benzoylecgonine(BZE) in urine samples. METHODS: A single dose of COC (10 mg/kg) was injected intraperitoneally into guinea pigs and urine samples were collected for 7 days. The urine samples were extracted by auto solid phase extraction (SPE), separated by the Allure PFP propyl column with a mobile phase consisting of methanol and 20 mmol/L ammonium acetate buffer [0.1% formic acid (80:20, V/V)], and then analyzed by LC-MS/MS. Multiple reaction monitoring (MRM) mode was used to analyze COC (m/z 304.2-->182.3, m/z 304.2-->150.1) and BZE (m/z 290.2-->168.3, m/z 290.2-->105.0). RESULTS: COC and BZE showed a fairly good linearity over the range of 2.0-100 ng/mL (r=0.9995). The detection limit was 0.5 ng/mL. The recovery rate was greater than 90% and the deviation of intra- and inter-day precision was less than 6%. BZE was the major target detected in urine samples, and its detection window was longer than COC. CONCLUSION This newly developed method shows high sensitivity and selectivity, and is suitable for the simultaneous analysis of cocaine and benzoylecgonine in urine samples.
Animals ; Chromatography, Liquid/methods* ; Cocaine/urine* ; Forensic Toxicology ; Guinea Pigs ; Humans ; Solid Phase Extraction ; Tandem Mass Spectrometry/methods*

Objective To investigate the clinical epidemiological features of femoral neck frac-tures. Methods The clinical data of patients with femoral neck fractures admitted to Third Affiliated Hospital of Hebei Medical University from 2003 to 2007 were retrospectively analyzed. All patients were divided into children group (age < 16 years) , adult group (age ranged from 16 to 60 years) and older group (>60 years). The types of the femoral neck fractures included 31-B1, 31-B2 and 31-B3 according to AO classification. The gender, age, fracture type and injury causes were analyzed. Results A total of 2 064 patients (971 males and 1 093 females) with femoral neck fractures were involved in the study.There were 356, 381, 397, 454 and 476 patients respectively in 2003, 2004, 2005, 2006 and 2007.There were 74 patients (3.59%) in the children group, 979 (47.43%) in the adult group and 1 011 (48.98%) in the older group. There were 960 patients (46. 51%) with type 31-B1 fractures, 860 (41. 67%) with type 31-B2 fractures and 244 (11. 82%) with 31-B3 fractures. Conclusions Form 2003 to 2007, the incidence of femoral neck fracture shows a trend of increase, with the highest incidence in the old persons. The male patients with femoral neck fractures are more than female patients in adult group, while the female patients with femoral neck fractures are less than male patients in older group.The dominant fractures type according to AO classification are type 31-B2 fractures in children and adult groups, but type 31-B1 fractures in older group.

A sensitive LC-MS/MS method to determine cocaine and its major metabolite benzoylecgonine in guinea pig' s hair has been established. About 20 mg of decontaminated hair sample was hydrolyzed with 0. 1 mol x L(-1) HCl at 50 degrees C overnight, in the presence of cocaine-d3 and benzoylecgonine-d8 used as internal standards, and then extracted with dichlormethane. The analysis was performed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Positive electrospray ionization (ESI +) and multiple reactions monitoring (MRM) mode were used. The limit of detection (LOD) for cocaine and benzoylecgonine was 1 pg x mg(-1). The calibration curves of extracted standards were linear over the range from 5 pg x mg(-1) to 250 pg x mg(-1) (r2 > or = 0.9997). The method was validated and applied to the analysis of guinea pig's hair after a single dose administration of cocaine hydrochloride. Cocaine and benzoylecgonine were not only detected, but also quantified in guinea pigs hair.
Animals ; Chromatography, High Pressure Liquid ; Cocaine ; administration & dosage ; analogs & derivatives ; analysis ; metabolism ; Guinea Pigs ; Hair ; chemistry ; metabolism ; Spectrometry, Mass, Electrospray Ionization ; Tandem Mass Spectrometry

AIMTo investigate the effect of brassinolide, a plant growth modulator, on multidrug resistance (MDR) of human T lymphoblastoid cell line CCRF-VCR 1000 which was obtained by progressively addition of vincristine (VCR) to sensitive CCRF-CEM cells, and to explore preliminarily the mechanism of reversing action.

METHODSMTT method was used to detect the resistant factor of resistant cell line and the reversing fold after addition of brassinolide. The intracellular accumulation of rhodamine 123, a fluorescent dye transported by P-glycoprotein was detected by flow cytometry, the catalytic activity of topoisomerase II was assessed by Sulliven method to find the effect of brassinolide on resistance. The protein expression of p53 was measured using Western blotting in the sensitive cells and resistant cells to explore the effect of brassinolide.

RESULTSThe resistant factors of CCRF-VCR cells on adriamycin, VP-16 and VCR are respectively as 153.1, 55.9 and 8123.1 folds comparing to the sensitive cell line CCRF-CEM. After treatment of brassinolide under the concentration of 0.001 - 10.0 microg x mL(-1), the resistance of CCRF-VCR was reversed partly with the reversing folds respectively as 4.4 - 11.6. The intracellular accumulation of rhodamine 123 was significantly reduced in the resistant cells. After treatment of brassinolide, the accumulation increased, the level of fluorescent dye was situated between resistant cells and sensitive cells. No alteration of the catalytic activity of topoisomerase II was found among three groups. The level of protein expression of p53 in resistant cells was higher than that of sensitive cells. After brassinolide treatment, the expression of p53 in CCRF-VCR cells restored to the level of sensitive cells.

CONCLUSIONBrassinolide could effectively reverse the resistance of CCRF-VCR cells by inhibiting the effusion of drug transported by P-glucoprotein. To down regulate the abnormal expression of p53 maybe one of the mechanisms of reversing MDR for brassinolide.

Brassica rapa ; chemistry ; Brassinosteroids ; Cell Line, Tumor ; drug effects ; Cholestanols ; isolation & purification ; pharmacology ; Drug Resistance, Multiple ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Humans ; Leukemia, T-Cell ; metabolism ; pathology ; Plant Growth Regulators ; pharmacology ; Pollen ; chemistry ; Steroids, Heterocyclic ; isolation & purification ; pharmacology ; Tumor Suppressor Protein p53 ; metabolism

Objective To derive the general equation of the probability distribution of identity by state （IBS） score among biological full sibling pairs by calculating STR allele frequency. Methods Based on the Mendelian genetics law and the hypothesis that parents of biological full siblings（FS） were unrelated individuals, the IBS score and corresponding probability of different genotype combinations in the offspring when unrelated individuals of different genotype combinations give birth to two offsprings were derived. Results Given fi （i=1, 2, …, m） as the frequency of the ith allele of a STR locus, the probability of sharing 2 alleles（p2FS）, 1 allele（p1FS） or 0 allele （p0FS） with biological full sibling pairs on the locus can be respectively expressed as follows： p2FS= 14 ×[1 + 2∑im= 1 fi2 + 2（∑im= 1 fi2）2 -∑im= 1 fi4] , p1FS= 12 ×[1 +∑im= 1 fi2 - 2（∑im= 1 fi2）2 - 2∑im= 1 fi3 + 2∑im= 1 fi4] and p0FS= 14 ×[1 - 4∑im= 1 fi2 + 2（∑im= 1 fi2）2 + 4∑im= 1 fi3 -3∑im= 1 fi4] . The sum of p2FS, p1FS and p0FS must be 1. As for the multiple genotyping system that contained n STR loci, IBS scores between biological full sibling pairs conform to binomial distribution：IBS~B（2n, π1）. The population rate π1, can be given by the formula：π1= 1n∑ln= 1 p2FSl + 21n∑ln= 1 p1FSl . Conclusion The alternative hypothesis in biological full sibling testing is that two appraised individuals are biological full siblings. The probability of the corresponding alternative hypothesis of any STR locus combination or IBS score can be directly calculated by the equations presented in this study, and the calculation results are the basis for explanations of the evidence.

OBJECTIVE: To investigate the feasibility of the new method of combining freeze grinding with ultrasonic technique for the pretreatment of the nail for toxicological and pharmaceutical analysis and to compare the advantages and disadvantages of this method with other traditional methods. METHODS: Five pretreatment methods were examined. Scanning electron microscope (SEM) was used to observe the microstructural changes of the nail. RESULTS: The microscopic structure of nail totally destroyed after alkali treatment. The hierarchy mode of the internal structure became obvious and tight after acid hydrolysis, which became indistinct after methanol infiltration. The structure of nail broke to pieces after ultrasonic technique. After freeze grinding combined ultrasonic technique, the particle structure kept original shape, and its size was one hundred times smaller than which after manual way. CONCLUSION The freeze grinding combined ultrasonic technique can improve the release efficiency, and ensure the stability of the toxicant or drug during the pretreatment process. It is appropriate for toxicological and pharmaceutical analysis in the nail.
Forensic Pathology ; Freeze Drying/methods* ; Humans ; Microscopy, Electron, Scanning ; Nails/chemistry* ; Particle Size ; Ultrasonics

Complete healing of the intestinal mucosa is the most ideal goal in the treatment of inflammatory bowel disease (IBD). The intestinal mucosa healing not only significantly alters the course of the disease and relieves clinical symptoms, but also markedly reduces the occurrence of complications and prevents recurrence of IBD. As chronic inflammation associated with peptic ulcer damage is the main pathological feature of IBD, clinical treatment is mainly based on anti-inflammatory therapy, but such therapy cannot promote the healing of the intestinal mucosa of patients. Therefore, how to achieve long-term remission of IBD is still an urgent challenge. In the process of intestinal mucosal repair, the polarization of macrophages maintains the homeostasis of the intestinal microenvironment, which is a representative process that promotes mucosal inflammatory-repair. It is a key part of initiating tissue regeneration that should not be underestimated. In this paper, we reviewed the literature of the past decade, focusing on the promotion of intestinal mucosal healing in IBD. The discussion will highlight the importance and feasibility of regulating macrophages to promote intestinal mucosal repair. Following this thought, we discuss the shortcomings of current clinical treatments and summarize the relevant drugs which have potential to promote intestinal mucosal repair. The aim is to provide effective potential drugs and therapeutic targets for the treatment of IBD.

The aim of this study was to investigate effects of IAT and MAT chemotherapeutic regimens treating patients with refractory and relapsed acute myeloid leukemia (AML). 99 patients with refractory and relapsed AML received IAT regimen or MAT regimen as study objects were retrospectively analyzed (56 patients with refractory AML and 43 patients with relapsed AML). Among of them, 28 patients were treated with IAT regimen, and 71 patients received with MAT regimen. The results showed that in 2 groups mentioned above the OR was 65.7%, CR was 49.5%, PR was 16.2%; in IAT group the OR was 64.3%, CR was 46.4%; in MAT group the OR was 66.2%, CR was 50.7%, no statistical difference was found between these 2 groups; The 2 years overall survival was 25% in IAT group and 15.5% in MAT group. Serious infection in IAT and MAT regime groups was 25% and 9.9%, respectively. It is concluded that both IAT and MAT regimens are effective methods for inducing CR in patients with refractory of relapsed AML. IAT and MAT regimens can be used in treatment of the refractory or relapsed MAL patients who were not respond to other regimen.
Adolescent ; Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; adverse effects ; therapeutic use ; Child ; Cytarabine ; administration & dosage ; Female ; Granulocyte Colony-Stimulating Factor ; administration & dosage ; Humans ; Leukemia, Myeloid, Acute ; drug therapy ; etiology ; Male ; Middle Aged ; Recurrence ; Retrospective Studies ; Vidarabine ; administration & dosage ; Young Adult