Objective:To establish an LC-MS/MS method for the content determination of triptolide in Shenshe ointment.Methods:The chromatographic separation was performed on an Agilent TC-C18 (250 mm×4.6 mm,5 μm)column with the mobile phase consisting of methanol-0.1% formic acid solution (73∶27,v/v).The flow rate was 0.5 ml·min-1 and the column temperature was maintained at 35℃.An electrospray ionization (ESI) source was applied with multiple reaction monitoring (MRM) combined with a positive mode.Metronidazole was used as the internal standard.The mass transitions were 361.4→43.2 for triptolide and 172.1→128.0 for metronidazole,respectively.Results:The linear range was 10-500 ng·ml-1 with good correlation coefficient (r=0.999 9).The limit of quantification (LOQ) was 9.5 ng·ml-1.The intra-and inter-day RSDs of peak areas were all less than 3% and the average recovery was 96.87%(RSD=2.79,n=6).Conclusion:The established LC-MS/MS method is simple,efficient,sensitive and accurate in the quality control of Shenshe ointment.

OBJECTIVETo study the changes of pharmacokinetics of 6,7-dimethoxycoumarin in a rat model of alpha-naphthylisothiocyanate (ANIT)-induced experimental hepatic injury after oral administration of Yinchenhao Decoction (, YCHD) using an ultra pressure liquid chromatography (UPLC) method.

METHODSRats were divided into a normal group and a model group, after modeled by 4% ANIT (75 mg/kg) for 48 h, they were orally administrated with YCHD extract at the dose of 0.324 g/kg, and then blood was collected from their orbital sinus after different intervals. Changes in liver function were monitored by the levels of liver enzymes [alanine aminotransferase (ALT), aspartate aminotransferase (AST)] and bilirubins [total bilirubin (TBIL), direct bilirubin (DBIL)], the concentration of 6,7-dimethoxycoumarin in plasma were measured by UPLC, and the pharmaceutical parameters were calculated with DAS2.1.1 software.

RESULTSThe concentration-time curve of both normal and modeled rats after oral administration of YCHD was obtained. Their time to maximum plasma concentration (t(max)) were both 0.25 h, the maximum concentration (C(max)) were 4.533 μg/mL and 6.885 μg/mL, and their area under concentration-time curve (AUC)(0→24h) were 16.272 and 32.981, respectively. There was a 51.88% and 100.46% increase in C(max) and AUC(0-t) (P<0.05), but there showed a 45.52% and 92.93% reduction in clearance of drug and volum of distribution (P<0.05), respectively.

CONCLUSIONSHepatic injury could significantly influence the pharmacokinetics of 6,7-dimethoxycoumarin after oral administration of YCHD, the absorption and distribution process was accelerated in liver injured rats, but the metabolism and elimination process was slowed. And this may lead to a significant accumulation of 6,7-dimethoxycoumarin in the body.

1-Naphthylisothiocyanate ; Administration, Oral ; Animals ; Chemical and Drug Induced Liver Injury ; blood ; drug therapy ; metabolism ; physiopathology ; Coumarins ; blood ; pharmacokinetics ; Disease Models, Animal ; Drug Evaluation, Preclinical ; Drugs, Chinese Herbal ; administration & dosage ; pharmacokinetics ; therapeutic use ; Liver ; drug effects ; physiopathology ; Models, Biological ; Rats ; Rats, Sprague-Dawley ; Validation Studies as Topic

OBJECTIVETo develop and investigate GLL-37, a substitution analogue of the human antimicrobial peptide LL-37 with anti-enzymatic degradation activity and improved efficacy.

METHODSThe bactericidal activities of LL-37 and newly developed GLL-37 against 6 Gram-negative and -positive bacteria were determined by Broth microdilution assays. The minimum inhibitory concentrations of LL-37 and GLL-37 against E.coli ATCC 25922 in different NaCl concentration medium were also detected. Both peptides were co-incubated with elastase, and then analyzed by PAGE electrophoresis and bactericidal activity determination.

RESULTGLL-37 showed a stronger elastase resistance ability than LL-37, and was significantly more effective than LL-37 under high-salt condition.

CONCLUSIONThe antimicrobial peptide GLL-37 derived form LL-37 has the potential as a new therapeutic agent for bacterial infections.

Animals ; Anti-Bacterial Agents ; pharmacology ; therapeutic use ; Antimicrobial Cationic Peptides ; pharmacology ; therapeutic use ; Blood Bactericidal Activity ; drug effects ; Cathelicidins ; Cell Membrane Permeability ; drug effects ; Escherichia coli ; drug effects ; Female ; Humans ; Membrane Proteins ; metabolism ; Monocytes ; drug effects ; Pseudomonas Infections ; drug therapy

OBJECTIVETo explore the crew and passengers' comfort on the Shijiazhuang-Taiyuan passenger dedicated line and physical factors, such as air pressure, noise, wind speed.

METHODSComfort investigation of all the crew (n = 244) and passengers (n = 377) on the Shijiazhuang-Taiyuan passenger dedicated line at speed of 250 km/h and 200 km/h and the detection of the air pressure, noise and wind speed were performed in 2011.

RESULTSSignificantly higher ratio of comfortable feeling, lower ratio of seriously discomfortable feeling were observed in crew and passengers at 200 km/h compared with those at 250 km/h (P < 0.05), as well as rapid disappearance of discomfortable feeling in crew (P < 0.05) and significantly higher ratio of lightly discomfortable feeling and lower ratios of tinnitus and eardrum discomfort induced by air pressure and noise in passengers at 200 km/h. No significant difference was observed in ear discomfort induced by air pressure and noise among crew, and the duration of disappearance of discomfortable feeling among passengers between 200 km/h and 250 km/h. The noise in carriages exceeded the related standard when the high-speed train passing through the tunnels.

CONCLUSIONThe individuals feel more comfortable at 200 km/h than 250 km/h in this line., which may be related with rapid variation of wind speed and noise when the train passes through the tunnels with high speed.

Adolescent ; Adult ; Atmospheric Pressure ; Female ; Humans ; Male ; Noise, Occupational ; Noise, Transportation ; Occupational Health ; Railroads ; Sensation ; Surveys and Questionnaires ; Wind ; Workplace ; Young Adult

Increasing evidence suggests that epigenetic dysfunction may influence the stability of normal pregnancy.The ten-eleven translocation (TET) family and 5-hydroxymethylcytosine (5-hmC) were found to be linked with epigenetic reprogramming.The present study aimed to examine the expression of the TET family and 5-hmC in the villi of human embryos and compared their expression between normal pregnancy and early pregnancy loss (EPL).Embryonic villi were collected from normal pregnant women (control) experiencing medical abortion and from EPL patients at gestation ages of 6,7 and 8 weeks.The mRNAs of TET family were analysed using quantitative polymerase chain reaction (qPCR),and TET proteins using Western blotting and immunohistochemical analysis.The MethylFlashTM Kit was used to quantify the absolute amount of 5-methylcytosine (5-mC) and 5-hmC.Our results showed that the expression of the TETs and 5-hmC in the normal villus decreased with increasing gestational age.Immunohistochemistry revealed that the TET proteins were expressed in the cytoplasm of trophoblasts and their expression was the highest in the 6-week tissue samples,which was consistent with the qPCR and Western blot results.The expression of TET1,TET2,and TET3 was lower in the villi in EPL group than in normal pregnancy group (P＜0.05 for all).It was concluded that the TET family and 5-hmC are critical in epigenetic reprogramming of human embryo.The findings also suggest that a deficiency of TETs in the villus might be associated with human EPL.

OBJECTIVE: To determine the relationship between intravoxel incoherent motion (IVIM) imaging derived quantitative metrics and serum soluble CD40 ligand (sCD40L) level in an embolic canine stroke model. MATERIALS AND METHODS: A middle cerebral artery occlusion model was established in 24 beagle dogs. Experimental dogs were divided into low- and high-sCD40L group according to serum sCD40L level at 4.5 hours after establishing the model. IVIM imaging was scanned at 4.5 hours after model establishment using 10 b values ranging from 0 to 900 s/mm². Quantitative metrics diffusion coefficient (D), pseudodiffusion coefficient (D*), and perfusion fraction (f) of ischemic lesions were calculated. Quantitative metrics of ischemic lesions were normalized by contralateral hemisphere using the following formula: normalized D = D(stroke) / D(contralateral). Differences in IVIM metrics between the low- and high-sCD40L groups were compared using t test. Pearson's correlation analyses were performed to determine the relationship between IVIM metrics and serum sCD40L level. RESULTS: The high-sCD40L group showed significantly lower f and normalized f values than the low-sCD40L group (f, p < 0.001; normalized f, p < 0.001). There was no significant difference in D*, normalized D*, D, or normalized D value between the two groups (All p > 0.05). Both f and normalized f values were negatively correlated with serum sCD40L level (f, r = −0.789, p < 0.001; normalized f, r = −0.823, p < 0.001). However, serum sCD40L level had no significant correlation with D*, normalized D*, D, or normalized D (All p > 0.05). CONCLUSION: The f value derived from IVIM imaging was negatively correlated with serum sCD40L level. f value might serve as a potential imaging biomarker to assess the formation of microvascular thrombosis in hyperacute period of ischemic stroke.
Animals ; CD40 Ligand* ; Diffusion ; Dogs ; Infarction, Middle Cerebral Artery ; Magnetic Resonance Imaging* ; Perfusion ; Stroke* ; Thrombosis

The aim of this study was to establish a method for quantitative detection of immunoglobulin heavy chain (IgH) gene rearrangements and to explore its clinical application in monitoring minimal residual disease (MRD) of acute lymphoid leukemia (ALL). Clonal IgH gene rearrangements in 51 patients with ALL at diagnosis were identified by multiplex PCR assay. PCR products were sequenced and pairwise in IMGT data base. Allele-specific oligonucleotides primers were designed complementary to the junctional region. The conservative JH primers combined with TaqMan probes were used to monitor the level of MRD in ALL patients. The sensitivity and specificity were assessed as well. The results indicated that out of all the 51 ALL patients, IgH rearrangements were identified in 21 cases, and 15 patients out of them were quantified. The slope of the standard curves was -3.1 to -3.9 and the correlation coefficients of all standard curves were > 0.98. The sensitivity was between 10(-4) and 10(-5), only one patient's sensitivity was 10(-3). Most of the quantitative range was less than 10(-4). The background's nonspecific amplification was detectable at a low level and had a little influence on results. 7 patients whose MRD level below 10(-3) kept complete remission and another 8 patients whose MRD level above 10(-3) had a higher relapse rate. It is concluded that the analysis of IgH gene rearrangements with RQ-PCR is a highly sensitive, specific and reliable method for accurate evaluation of MRD in ALL. The data indicates a high correlation between the level of IgH rearrangements and the prognosis in ALL patients.
Alleles ; DNA Primers ; genetics ; Female ; Humans ; Immunoglobulin Heavy Chains ; genetics ; Male ; Neoplasm, Residual ; diagnosis ; genetics ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; diagnosis ; genetics

This article describes the preparation of the N-tert-butyl-alpha-phenylnitrone (PBN) liposomes and their related characteristics. The PBN liposomes were prepared by film dispersion-supersonic method and the formula of liposomes was optimized by orthogonal uniform design. RP-HPLC was used to qualify the amount of PBN that entered into the hepatoma cells. Necrosis rate was also investigated by fluorescence activated cell sorter (FACS) after PBN liposomes transfection. Result showed that the mean particle size, entrapment efficiency, and polydispersity of the resulting PBN-liposome were 137.5 nm, 71.52% and 0.286, respectively. PBN liposomes can enter into the tumor cell stably and they have higher affinity to hepatoma cell compared with free PBN resulting in a higher necrosis rate after transfection. These results provide a potential method for early diagnosis and treatment of cancer using specific spin trapping probe targeting tumor cells.
Carcinoma, Hepatocellular ; pathology ; Cyclic N-Oxides ; chemistry ; Electron Spin Resonance Spectroscopy ; Humans ; Nanoparticles ; chemistry ; Spin Labels ; Spin Trapping ; methods ; Tumor Cells, Cultured

OBJECTIVETo summarize the clinical data in preventing HBV recurrence after liver transplantation and explore a optimal individual protocol in prophylaxis of HBV recurrence.

METHODSWe retrospected outcomes in 195 recipients who underwent a liver transplantation for HBV-related liver disease between June 2004 and July 2008. According to the anti-virus protocol these recipients are divided into two groups as following: group A received a protocol of combination treatment of lamivudine with HBIG, and group B with combination treatment of adefovir with HBIG. With mean follow-up of 23.7 months, HBV recurrent rate was observed in overall and each group separately.

RESULTSA total of 195 liver transplant recipients were identified that met the study criteria. At the sixth and eleventh month after operation, HBV recurrence appeared in 2 recipients, each in two groups, which were due to LAM cessation and HBV mutation respectively. Recurrent rate was 0.6% in group A, 3.7% in group B and 1% in total. There was no significant difference in HBV recurrent rate between group A and B.

CONCLUSIONLamivudine combined with HBIg should be considered as a reliable method in preventing HBV recurrence after liver transplantation. Better outcomes can be achieved by individual anti-virus protocol and HBIg administration according to HBV status in recipient.

Adolescent ; Adult ; Aged ; Antibiotic Prophylaxis ; Antiviral Agents ; therapeutic use ; Female ; Hepatitis B ; prevention & control ; surgery ; Hepatitis B virus ; drug effects ; physiology ; Humans ; Lamivudine ; therapeutic use ; Liver Transplantation ; Male ; Middle Aged ; Recurrence ; Young Adult

BACKGROUNDMinimally invasive surgery in skull base relying on searching for possible anatomic basis for endoscopic technology is controversial. The objective of this study was to observe the spatial relationships between main blood vessels and nerves in the cerebellopontine angle area and provide anatomic basis for lateral and posterior skull base minimally invasive surgery via endoscopic retrosigmoid keyhole approach.

METHODSThis study was conducted on thirty dried adult skulls to measure the spatial relationships among the surface bony marks of posterior cranial fossa, and to locate the most appropriate drilling area for retrosigmoid keyhole approach. In addition, we used 10 formaldehyde-fixed adult cadaver specimens for simulating endoscopic retrosigmoid approach to determine the visible scope.

RESULTSThe midpoint between the mastoid tip and the asterion was the best drilling point for retrosigmoid approach. A hole centered on this point with the 2.0 cm in diameter was suitable for exposing the related structures in the cerebellopontine angle. Retrosigmoid keyhole approach can decrease the pressure on the cerebellum and expose the related structures effectively which include facial nerve, vestibulocochlear nerve, trigeminal nerve, glossopharyngeal nerve, vagus nerve, accessory nerve, hypoglossal nerve, anterior inferior cerebellar artery, posterior inferior cerebellar artery and labyrinthine artery, etc.

CONCLUSIONSExact location on endoscope retrosigmoid approach can avoid dragging cerebellum during the minimally invasive surgery. The application of retrosigmoid keyhole approach will extend the application of endoscopic technology.

Adult ; Cerebellopontine Angle ; anatomy & histology ; surgery ; Endoscopes ; Humans ; Minimally Invasive Surgical Procedures