OBJECTIVE: To establish the method to analyze γ-hydroxybutyric acid (GHB) and its precursors 1,4-butanediol (1,4-BD) and gamma-butyrolactone (GBL) in urine through LC-MS/MS and provide evidence for related cases. METHODS: GHB-d6 and MOR-d3 were used as the internal standard. The urine sample was separated by LC after protein precipitation with methanol. The electrospray ion source was for ionization. Each compound was detected through multiple-reaction monitoring (MRM) mode. RESULTS: The limits of detection of GHB and its precursors 1,4-BD and GBL were 0.1, 0.1 and 2 μg/mL. The accuracy was 87.6%-98.1%. The intra-day and inter-day precisions were less than 15% and matrix effects were higher than 80%. CONCLUSION The method is high sensitive, simple, rapid, specific and with high reliability. This study has provided technical support and basic data for forensic cases involving GHB.
4-Butyrolactone/urine* ; Butylene Glycols/urine* ; Chromatography, Liquid ; Forensic Sciences ; Humans ; Hydroxybutyrates/urine* ; Mass Spectrometry ; Reproducibility of Results ; Tandem Mass Spectrometry

BACKGROUND:Cytokines and neurotrophic factors secreted from human umbilical cord blood-derived mesenchymal stem cells secrete have neuroprotective effects on cerebral ischemia-reperfusion injury, but there are few reports about intranasal administration of human umbilical cord blood-derived mesenchymal stem cellconditioned medium in the treatment of stroke.
OBJECTIVE:To investigate the protective effects of intranasal administration of human umbilical cord-derived mesenchymal stem cells-conditioned medium on neurologic function of rats with cerebral ischemia-reperfusion injury.
METHODS:Adult rats were subjected to 2 hours of right middle cerebral artery occlusion and the human umbilical cord-derived mesenchymal stem cells were isolated from the postpartum human cord. We made the conditioned medium of human umbilical cord-derived mesenchymal stem cells. Ischemic rats were randomized and assigned to three groups and were treated by intranasal routine starting 24 hours after middle cerebral artery occlusion with:(1) saline for control group;(2) Dulbecco’s modified Eagle’s medium/Ham’s nutrient mixture F-12 medium for medium control group;(3) conditioned medium treatment group (10mL/kg) daily for 14 days. Behavioral tests (foot fault test, and modified Neurological Severity Score) were performed before and at 1, 7, 14 days after middle cerebral artery occlusion.
RESULTS AND CONCLUSION:There was no difference in the behavioral tests among the three groups at postoperatively 1 day (P>0.05). Compared to the control and medium control group rats, respectively, rats in the conditioned medium group significantly improved functional outcome after stroke in days 7 and 14 (P<0.05). There was also no significant difference in functional tests between the control group and medium control group in days 7 and 14 (P>0.05). These results suggest that human umbilical cord-derived mesenchymal stem cells-conditioned medium via intranasal administration can significantly improve neurologic functional outcome after cerebral ischemia-reperfusion injury.

Objective To investigate the feasibility and accuracy of magnetic resonance microneurog- raphy of sciatic nerve fascicles in rabbil by correlation with the gross anatomy.Methods The 3D T_2-weigh- ted imaging(3D-T_2 MI),3D T_2-weighted imaging plus spectral presaturation with inversion recovety(SPIR), T_1-weighted imaging(T_1 WI)of the sciatic nerve in 10 rabbits were performed on a 1.5 Tesla magnetic reso- nance system.The radiological ananomy and imaging features of sciaticnerve fascicles were observed and the anterior-posterior diameter was measured on 3D T_2-weighted imagingThe imaging evaluation was correlated with the gross anatomy.The T_1 and T_2 relaxation time were determined by multiple echo spin echo and mix se- quecerespectively.Results The libial nerve and peroneal nerve in the main trunk of sciaticnerve in all 10 rabbits could be clearly displayed on the 3D T_2 WI,3DT_2WI plus SPIRand T_1WI.Strikingly,the 3D T_2 WI could delineate the fine branches of the sural nerve and posterior femural cutaneous nervesThe T_1 and T_2 relaxation time were 915 ms40 msrespectivelyGrosslythe anterior-posterior diameter of sciatic nerve trunk was3.17?0.21)mmwhile was(3.15?0.19)on 3D T_2 WI.There was no statistically significant difference(t=0.768,P=0.462).Conclusion With 1.5 Telsa MR system the microneurography of the sci- atie nerve could be achievable and the individual fascicles of sciatic nerve trunk could be clearly and accurately discriminated.

Objective To investigate the effects of partial body weight supported treadmill training (BW- STT) on post-stroke depression (PSD) and on patients' quality of life.Methods Sixty patients with PSD were re- cruited and divided into a training group (n=30,male 17,female 13) and a control group (n=30,male 16,fe- male 14).All patients were treated with routine internal medication and rehabilitation.The patients of the training group also received BWSTT in addition to their routine treatment.All patients' neurological impairment was evaluated using the Modified Edinburgh-Scandinavian Stroke Scale (MESSS).The Hamilton depression scale (HAMD) was used for evaluating the degree of depression.The Fugl-Meyer scale and the Barthel index were used to assess ambula- tion and balance,and facility in the activities of daily living.All patients were assessed before and after the treat- ment.Results After four weeks of treatment,depression in the training group had improved significantly more than in the control group.Conclusion BWSTT intervention is very important for patients with PSD:it can reduce the degree of depression and improve the quality of life.

Objective The purpose of this study was to investigate the clinical significance of serum vascular endothelial growth factor(S-VEGF)in patients of esophageal squamous cell carcinoma with the end-point of progress-free survival rate.Methods Sera from 89 patients with esophageal squamous carcinoma,who presented and treated with concurrent chemoradiotherapy or surgical resection in Cancer Center d Sun Yat-Sen University from December 2002 to May 2004,were sampled at the time of pre-treatment.30 cases of health individuals without any evidence of disease were selected as control group.For patients with surgical resection were performed with esophagectomy and extended lymphadenectomy.For patients with concurrent chemoradiotherapy comprised of cisplatin and 5-fluorouracil.The radiotherapy dose of 2 Gy per day was initiated on Day 1 of chemotherapy and continued daily for 5 days per week for 6 weeks,for a total close of 60 Gy.Two courses of chemotherapy were given during radiotherapy at 6-week intervals.The serum vascular endothelial growth factor(S- VEGF)levels were measured with a solid phase enzyme Human VEGF Immunoassay ELISA kit.The end point of this study was progress-free survival,and time was calculated from the date of diagnosis to date of relapse or last follow evaluation.Results S-VEGF level of patients with esophageal squamous cell carcinoma was significantly higher than that of heahhy controls(475.93?44.76 pg/ml vs.294.20?23.40 pg/ml;P=0.020).S-VEGF levels in patients with StageⅢand StageⅣdisease were significantly higher than those in patients with Stage I and StageⅡdisease.The 1-year progress-free survival rate d high S-VEGF group(≥475 pg/ml)was significantly lower than that of the low S-VEGF group(＜475 pg/ml)(24% vs.66%;P=0.0004).The 1-year progress-free survival rate of the patients with StageⅠand StageⅡdisease was significantly higher than that of patients with StageⅢand StageⅣdisease(71% vs.29%;P=0.0015).The variables were assessed by multivariate analysis using the Cox proportional hazards model,and the results revealed that the clinical stage and the S-VEGF were first and second independent prognosis factor,respectively.Conclusions In the current study,a high S-VEGF was found to be associated with tumor progression,poor treatment response,and poor survival in patients with squamous cell carcinoma of the esophagus.

ObjectiveTo investigate the effect of FUT8-siRNA on transforming growth factor β (TGF-β)-Smad2/3 signalling pathway in renal tubular epithelial cells.MethodsHK-2 cells were divided into six groups:normal group,negative control group,TGF-β1 group,TGF-β1 with FUT8 interference group,TGF-β1 with negative control group,FUT8 interference group.RNAi was performed to silence the expression of FUT8 gene,then immunofluorescent analysis was used to detect the expression of core fucose in the HK-2,immunoprecipitation and lectin blotting were performed to detect the core fucosylation of TGF-βR Ⅱ and ALK-5,and detect the change of Smad2/3 and p-Smad2/3 in HK-2 cells after FUT8 gene was silenced.ResultsCompared with the normal and negative control group,incubation with 5 μg/L TGF-β1 for 48 h could significantly up-regulate the core fucosylation of HK-2 cells,enhance the protein expression of TGF-βR Ⅱ and ALK-5 (P＜0.05),markedly increase the expression level of p-Smad 2/3 (P＜0.05) and cause it to nuclear translocation in HK-2 cells.While FUT8siRNA could inhibit the above up-regulation of TGF-βR Ⅱ and ALK-5(P＜0.05),suppress the increase of p-Smad 2/3(P＜0.05) and its nuclear translocation without disturbing the protein expression of TGF-βR Ⅱ and ALK-5.Conclusion FUT8-catalized core fucosylation of TGF-βR Ⅱ and ALK-5 is needed to fulfill their functions,and blocking core fucosylation of TGF-βR Ⅱ and ALK-5 leads to the inhibition of TGF-β-Smad2/3signalling pathway in HK-2 cells.

Two trace impurities in the bulk drug lisinopril were detected by means of high-performance liquid chromatography coupled with mass spectrometry (HPLC/MS) with a simple and sensitive method suitable for HPLC/MSn analysis. The fragmentation behavior of lisinopril and the impurities was investigated, and two unknown impurities were elucidated as 2-(6-amino-1-(1-carboxyethylamino)-1-oxohexan-2-ylamino)-4-phenylbutanoic acid and 6-amino-2-(1-carboxy-3-phenylpro-pylamino)-hexanoic acid on the basis of the multi-stage mass spectrometry and exact mass evidence. The proposed structures of the two unknown impurities were further confirmed by nuclear magnetic resonance (NMR) experiments after preparative isolation.
Chromatography, High Pressure Liquid ; methods ; Drug Contamination ; Lisinopril ; analysis ; Magnetic Resonance Spectroscopy ; Spectrometry, Mass, Electrospray Ionization ; methods

BACKGROUND:Large numbers of experimental data have confirmed that bone marrow mesenchymal stem cel s have a positive therapeutic effect on cerebral ischemia-reperfusion injury, but there are few reports about intravenous administration of bone marrow mesenchymal stem cel conditioned medium in the treatment of stroke.
OBJECTIVE:To investigate the effects of the conditioned medium of rat bone marrow mesenchymal stem cel s on the recovery of neurological function in rats after cerebral ischemia-reperfusion injury.
METHODS:The bone marrow mesenchymal stem cel s were isolated from rat bone marrow. When cel s at passage 2 or 3 reached 90%confluence, the original culture medium was removed. Then the cel s were cultured in serum-free DMEM for 18 hours. After that, the culture solution was col ected as the conditioned medium of rat bone marrow mesenchymal stem cel s. Adult rats were subjected to 2 hours of right middle cerebral artery occlusion. Ischemia-reperfusion injury rats were randomly assigned to three groups:control group, simple culture medium group and conditioned medium group, and respectively given injection of normal saline, DMEM, conditioned medium (10 mL/kg) via the tail vein at 2, 24, 48 hours after operation.
RESULTS AND CONCLUSION:There was no difference in the behavioral tests among the three groups at postoperative 2 hours (P>0.05). Compared with the control and simple culture medium group, neurological impairment was significantly improved in the conditioned medium group at postoperative 1, 3, 5 days (P<0.05), but there was no significant difference between the control and simple culture medium groups. At postoperative 5 days, brain edema was significantly eased in the conditioned medium group in comparison to the control and simple culture medium groups (P<0.05), and there was also no difference between the latter two groups (P>0.05). These results suggest that rat bone marrow mesenchymal stem cel s-conditioned medium via intravenous administration can significantly ease brain edema and improve the neurologic function after cerebral ischemia-reperfusion injury.

Objective To explore the clinical efficacy and safety of low-dose decitabine combined with cytarabine for myelodysplastic syndromes (MDS). Methods Clinical data of 15 patients with MDS who took the therapeutic regimen with decitabine combined with cytarabine were collected from January 2012 to January 2015. The clinical efficacy and adverse effects were assessed. Results Among the 15 patients, 4 cases were complete remission (CR), 5 cases were partial remission (PR) and 6 cases were stable disease (SD) and progressive disease (PD). The total effective rate was 60.0 % (9/15). Grade Ⅲ-Ⅳ bone marrow depression occurred in 11 cases with incidence rate of 73.3 % (11/15), and the total incidence rate of infection was 40.0 % (6/15), including lung infection of 26.7 % (4/15). All the infections were controlled after active supportive treatment and anti-infection therapy. No patient died of chemotherapy. Conclusions Low-dose decitabine combined with cytarabine can effectively treat MDS and delay the progress of disease. The patients can tolerate the adverse effects in chemotherapy with a low mortality rate.

Objective To explore the clinical effect and toxicity of daunorubicin combined with cytarabine (DA regimen) and idarubicin combined with cytarabine (IA regimen) for the treatment of patients with acute myeloid leukemia (AML) as induction chemotherapy. Methods The clinical data of 84 newly diagnosed AML patients (except M3) treated with DA or IA regimen were analyzed retrospectively. DA regimen group included 32 patients (17 males and 15 females with median age of 46 years), while IA regimen group included 52 patients (29 males and 23 females with media age of 49 years). Efficacy index was complete remission (CR), total efficiency and adverse reactions after one course of chemotherapy rate. Results In DA regimen group,the CR rate was 65.6 %(21/32), and the total efficiency rate was 75.0 %(24/32), while in IA regimen group, the CR rate was 71.2 %(31/52), and the total efficiency rate was 80.8 %(42/52), respectively, but, the differences of media survival and 5-year survival rate were not statistically significant (16.8 months vs. 24.9 months, 26 % vs. 44 %, both P>0.05). The main side effect in the two groups included hematologic (bone marrow suppression) and non-hematologic adverse reactions, with no significant difference between the two groups (all P>0.05). Conclusion For newly diagnosed AML patients, remission rate and total efficiency of DA regimen are same as IA regimen after one course treatment, and adverse events between the two regimens do not differ significantly.