The biotransformation, CYP reaction phenotyping, the impact of CYP inhibitors and enzyme kinetics of 3-cyanomethyl-4-methyl-DCK (CMDCK), a new anti-HIV preclinical candidate belonging to DCK analogs, were investigated in human intestinal microsomes and recombinant cytochrome P450 (CYP) enzymes. CMDCK (4 micromol L(-1)) was incubated with a panel of rCYP enzymes (CYP1A2, 2C9, 2C19, 2D6 and 3A4) in vitro. The remaining parent drug in incubates was quantitatively analyzed by a LC-MS method. CYP3A4 was identified as the principal CYP isoenzyme responsible for its metabolism in intestinal microsomes. The major metabolic pathway of CMDCK was oxidation and a number of oxidative metabolites were screened with LC-MS. The Km, Vmax, CLint and T1/2 of CMDCK obtained from human intestinal microsome were 45.6 micromol L(-1), 0.33 micromol L(-1) min(-1), 12.1 mL min(-1) kg(-1) and 25.7 min, respectively. Intestinal clearance of CMDCK was estimated from in vitro data to be 3.3 mL min(-1) kg(-1), and was almost equal to the intestinal blood flow rate (4.6 mL min(-1) kg(-1)). The selective CYP3A4 inhibitors, ketoconazole, troleandomycin and ritonavir demonstrated significant inhibitory effects on CMDCK intestinal metabolism, which suggested that co-administration of CMDCK with potent CYP3A inhibitors, such as ritonavir, might decrease its intestinal metabolic clearance and subsequently improve its bioavailability in body.

Depending on Shanghai medical big data center and taking the medical big data after quality control and before data utilization as research object,the paper establishes the data cleaning frame,gives the evaluation method for data availability,finds out the corresponding cleaning strategies according to the clustering analysis of data characteristics and repeatedly deduces the accuracy,reliability of the strategy,thus providing a strong support for the analysis and utilization of medical big data.

Objective To investigate the potential relationship between initial 131Ⅰ uptake by lung metastases from differentiated thyroid cancer (DTC) and treatment outcomes. Methods From 1997 to 2009, 41 patients with DTC lung metastases were treated in the authors' department. 131Ⅰ whole body scan (WBS), serum Tg levels and other imaging results were analyzed to evaluate the therapeutic effects. Complete response (CR) or partial response (PR) was considered to be effective. The x2 test and correlation analysis were performed using SPSS 11.5 software package. Results 131 Ⅰ treatment was effective in 63% (26/41) patients with DTC lung metastases, CR in 8 patients and PR in 18 patients. In other 37% ( 15/41 ) patients, 131Ⅰ treatment was ineffective, including one case died of distant metastases. Patients with initial presence of 131Ⅰ lung uptake had higher effective rate than those with 131Ⅰ lung uptake during the second or later 131Ⅰ treatment (76% (22/29)vs33% (4/12),x2 =4.911, P=0.027). Also, significantly higher effective rate was found in patients with lung metastases alone than those with extra-pulmonary metastases (75% (24/32) vs 22% (2/9), x2 = 6. 312, P =0.012). However, the effective rate in patients with diffuse metastases was not significantly different from that in patients with focal metastases (67% (12/18)vs 61% ( 14/23), x2 =0. 146, P=0.702). The positive rate of initial 131Ⅰ uptake by lung metastases was higher in patients with total thyroidectomy than those with partial thyroidectomy (83% (24/29) vs 42% (5/12) ). Those positive rates in patients with papilary DTC and patients with follicular DTC were 72% (23/32) and 6/9, respectively. The surgical mode was correlated with the initial 131Ⅰ uptake by lung metastases (r = 0.411, P ＜ 0.05), but no correlation was found between the histological type and the initial 131Ⅰ uptake by lung metastases ( r = 0. 047, P ＞ 0.05 ). Conclusion Initial uptake of 131 Ⅰ by lung metastases alone is a favorable prognostic factor for DTC patients treated by131Ⅰ, and total thyroidectomy may be beneficial for initial 131Ⅰ uptake by lung metastases.

Cell Transformation, Neoplastic ; drug effects ; genetics ; Dimethyl Sulfoxide ; pharmacology ; Electroporation ; methods ; HL-60 Cells ; Humans ; Membrane Proteins ; genetics ; Neoplasm Proteins ; genetics ; RNA Interference ; RNA, Small Interfering ; genetics ; Stromal Interaction Molecule 1 ; Transfection

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.

A high activity isoflavone-glucosidase, which hydrolysis glycosides, was obtainde using liquid fermentation from Absidia sp. R strain. The isoflavone-glucosidase was purified 11 folds with yielding rate of 10.9% after ammonium sulfate precipitation and DEAE-Cellocuse (DE-52) ion exchange chromatography. SDS-PAGE results showed that the molecular weight is 53kD. And the optimum temperature, the optimum pH, Km and pI of the enzyme are 50 deegrees C, 5.0, 1.3 x 10(-2) mol/L and 3.2, respectively. The isoflavone-glucosidase is also rather stable under 60 degrees C and in pH range from 5.0 to 7.0. The enzyme can be activated by Co2+ and Ca2+, and be inhibited by Ag+ and Cu2+.
Absidia ; enzymology ; Glucosidases ; isolation & purification ; metabolism ; Hydrogen-Ion Concentration ; Isoflavones ; metabolism ; Temperature

ObjectiveTo investigate the clinical effects and side-effect of mesenchymal stem cell(MSCs) transplantation on spinal cord injury(SCI),traumatic brain injury(TBI),multiple sclerosis(MS) or Parkinson's disease(PD).MethodsThe bone marrow(222～350 ml) of 11 patients with SCI(n=6),TBI(n=3),MS(n=1) or PD(n=1) were harvested from the patients' ilia and then MNCs were isolated.The MNCs were injected intravenously or into subarachnoid space by lumbar puncture.The neural function and side-effect were observed before and after MSCs transplantation and the patients were followed up.ResultsThe data demonstrated the improvement of sense and motor function in 5 patients with SCI,one had no improvement by 2 months following-up.These patients' sense and motor levels improved obviously.Their muscle strength of lower extremity increased,the muscular tone decreased and urinary bladder function improved.Changes in neurological deficits and improvements in function may appear within 2 days after transplantation,most of them within 2 weeks.There were significantly amelioration in 3 patients with TBI treated with MSCs transplantation,one of them could walk with cane independently after 3 months.One's PVS score elevated from 5 to 8 scales after transplantation.The tremor was alleviated after 1 week,and the muscular tone decreased,which lead to reduce the dose of Madopar after 3 months,in patient with PD.The patient with MS showed no improvement in short time.The side-effect included fever(7/11),headache(2/11) and abdominal dissension(1/11).1 patient feel numb in his legs while injection into subarachnoid,and appeared meningeal stimulation after injection.ConclusionThere were significantly clinical effects in treatment of SCI,TBI,MS,and PD with MSCs transplantation in short time,and with few side-effect. The long-term clinical effects need more observation with larger samples.

OBJECTIVETo compare the sensitivity and reproducibility of Allglo and TaqMan probe in the detection of simian immunodeficiency virus (SIV) using fluorescence quantitative RT-PCR (QPCR).

METHODSThe reference sample of SIV was diluted to 6 gradient concentrations; at each concentration 12 samples were tested to analyze the variations within batches, and each sample was tested for 12 times for analysis of variations between batches by QRT-PCR using TaqMan probe and Allglo probe. The results of QPCR using the two probes were analyzed with ABI7300 PCR system software.

RESULTSIn QPCR using TaqMan and Allglo probe, the lower limit of sensitivity for SIV detection was both 50 copies/mL. Assessment of the reproducibility of the tests showed that the maximum and minimum coefficients of variation between batches were 0.63% and 0.33% with Allglo probe, respectively, as compared with 1.33% and 0.2% with TaqMan probe. The maximum and minimum coefficients of inter-batch variation was 1.77% and 0.95% with Allglo probe, respectively, as compared with 1.86% and 1.03% with TaqMan probe.

CONCLUSIONAllglo probe shows a better performance then TaqMan probe in detection of SIV QPCR.

AIMTo investigate the effects of a nutritional supplement on nutritional status and hypoxia endurance in young adults living at high altitude.

METHODSForty healthy male young adults were recruited and randomly assigned to control and intervention groups. The nutrition survey was carried out using weighing method. The intervention group was given a nutritional supplement specifically designed for use at high altitude, while the control group was treated with a supplement made of stir-fried flour. After 20 days of supplementation, they marched from the altitude of 3700 m to 5100 m. The changes in HR, SaO2, serum concentrations of VA and VB2 and some minerals were measured.

RESULTSThe results of nutrition survey showed that the ratio of three macronutrients was not adequate and the intakes of calcium, VA and VB2 were below Chinese RNI. The serum concentrations of calcium, magnesium and VA were below normal references. The serum VB2 concentration was at the low level o f normal reference. The nutritional supplement could increase the serum concentrations of calcium, magnesium, VA and VB2, indicating an improved nutritional status. The changes in HR and SaO2 were diminished in intervention group compared with control group.

CONCLUSIONThe nutritional supplement can improve nutritional status and increase the hypoxia endurance in young adults living at high altitude.

Adult ; Altitude ; Dietary Supplements ; Humans ; Hypoxia ; prevention & control ; Male ; Nutritional Status ; Vitamins ; therapeutic use

OBJECTIVETo test the feasibility of correcting conjunctival sac narrowing following orbital implantation using polyester fiber heart patches instead of the skin autograft.

METHODSTwelve patients of conjunctival sac narrowing after orbital implantation (including 3 with orbital implant exposure) admitted in Nanfang Hospital between 2012 and 2016 received surgical correction of the conjunctival sac using polyester fiber heart patches. During the surgery, the central conjunctival sac was opened, the exposed area was covered with suitable polyester fiber heart patches, and the palpebral margin was sutured.

RESULTSThree months after the operation, 10 patients showed improved appearance after implantation of the prosthetic eye. Two patients received a second operation to remove the patches due to graft rejection and infections and skin autograft was implanted for reconstruction of the conjunctival sac.

CONCLUSIONPolyester fiber heart patches are ideal materials for repairing Conjunctival sac narrowing and orbital implant exposure, but this approach is not suitable in cases of severe narrowing or occlusion of the conjunctival sac.