Medicine, Chinese Traditional ; methods ; trends

Coronary Disease ; diagnosis ; Humans ; Medicine, Chinese Traditional ; methods

Objective To improve the level of diagnosis and treatment of acute appendicitis in leukemia.Method 17 cases of acute appendicitis in leukemia that were treated from 1980 to 2002 were reviewed.Result All 17 cases underwent appendectomy,16 survived the immediate postoperative period,only one died.Conclusions Leukemia were predisposed to acute appendicitis than normal during chemotherapy period and our experience supports the surgical management of acute appendicitis in leukemia.Because immunosuppression and thrombocytopenia were the main causes of additional surgical dangers,relative methods should be applied in perioperation period.The complications of incision were common,so the observation of incision should be emphasized.

Apoptosis ; Carcinoma, Squamous Cell ; diagnosis ; genetics ; metabolism ; therapy ; Cell Proliferation ; Chromosome Aberrations ; Early Diagnosis ; Epigenesis, Genetic ; Humans ; MicroRNAs ; genetics ; metabolism ; Mouth Neoplasms ; diagnosis ; genetics ; metabolism ; therapy ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Polymorphism, Genetic

In the process of evolution, pathogenic Streptococcus pyogenes secretes an immunoglobulin G-degrading enzyme IdeS which can specifically cleave the hinge region of immunoglobulin G in order to escape the immune response against the host. On the one hand, IdeS can be used for IgG fingerprinting as a tool enzyme combined with mass spectrometry technology. On the other hand, IdeS can be used to treat the antibody-responsive diseases produced by autoimmunity as a therapeutic protein. In this study, the backbone of plasmid pCold was used to construct two expression vectors of recombinant protein IdeS, which were heterologously expressed in Escherichia coli Shuffle T7. After purification by affinity chromatography, the recombinant IdeS activity was detected and their activity differences between the two were compared. Among them, the yield of the recombinant IdeS containing the His6-tag at the N-terminus was 4 mg·L^-1, and the cleavage reaction with antibody IgG1 at 1∶200 (m/m) at 37 ℃ for 30 min could complete. However, the yield of the recombinant IdeS containing both the N-terminal His6 tag and the C-terminal silica affinity tag (silica bing peptide, SiBP) is 1.5 mg·L^-1, and the degradation reaction with antibody IgG1 at 1∶20 (m/m) at 37 ℃ for 30 min could reach the end. The C-terminal fusion peptide has a great influence on the yield and activity of IdeS, which is not conducive to subsequent application in drug development. Above all, the recombinant IdeS containing the His6-tag at the N-terminus expressed by this system has high activity and can fully meet the needs of antibody drug development and mapping analysis of IgG.

Abstract?AlM: To investigate the effect of zebularine ( Zeb ) loaded Poly ( ethylene glycol ) - block - poly ( ε -caprolactone) methyl ether ( MePEG-PCL) nanoparticles ( NPs) on the viability, attachment, and apoptosis of in vitro cultured lens epithelial cells ( LECs) .?METHODS: In vitro cultured infant human lens tissue HLE B-3 immortalized cells were distributed randomly divided into six groups. Each group was administered with free Zeb 50μmol/L ( ZebF1 group ) , 100μmol/L ( ZebF2 group) , Zeb -loaded MePEG-PCL NPs 50μmol/L ( ZebNP1 group) , Zeb -loaded MePEG-PCL NPs 100μmol/L ( ZebNP2 group) , MePEG-PCL empty NPs( NPs group) or blank medium (group C) respectively. A tetrazolium dye assay ( MTT) test and modified MTT test were performed to determine cell viability and cell attachment. DNA ladder was used to detect the cell apoptosis.?RESULTS: Determined by MTT colorimetric method:Cell proliferation rate of LECs were suppressed by all Zeb administration groups in a concentration-time dependent manner (P<0. 05). Compared with the free Zeb groups, the viability of LECs were suppressed more effectively by the same dose of Zeb loaded MePEG-PCL NPs after 24, 48, 96h ( P < 0. 05 ). Determined by improved MTT colorimetric method: The attachment of LECs were decreased in all Zeb administration groups, Zeb loaded MePEG-PCL NPs had better effect on suppressing the attachment of LECs than the free Zeb groups with same dose (P<0. 05). The DNA ladder confirmed that after administration of 96h, group C, NPs group and ZebF1 group showed no DNA fragment, however the DNA fragment were performed in ZebF2, ZebNP1, ZebNP2 groups and displays the trend of ZebNP2> ZebNP1>ZebF2 (P<0. 05).?CONCLUSlON: Zeb loaded MePEG-PCL NPs had better effect on suppressing the viability and attachment of in vitro cultured LECs than the free Zeb groups , as well as enhancing the apoptosis.

AlM:To observe the effects of double incision combined surgery of single - stab trabeculectomy and phacoemulsification.
METHODS: Totally 28 cases ( 30 eyes ) with glaucoma and cataract undertook the modified combined surgery of single - stab trabeculectomy and phacoemulsification. After traditional phacoemulsification, cut the bulbar conjunctiva and Tenons capsule from the 11 o'clock to 1 o'clock, then puncture into the anterior chamber in 2mm behind the corneal limbus with 3mm tunnel knife, shaping a 3mm wide, 1/3-1/4 thickness scleral tunnel. Getting into the trabecular tunnel, bite off 3 pieces of trabecular tissue about 1mm × 1mm size. The changes in the imtraocular pressure ( lOP ) and the visual acuity before and after the surgery as well as filtering bleb ( OCT confirmed) and complications were carefully observed in 3-6mo postoperatively.
RESULTS: The postoperative visual acuity in 1wk postoperatively less than 0. 1 was found in 3 eyes, from 0. 1 to 0. 3 was found in 6 eyes,from 0. 3 to 0. 6 in 13 eyes, from 0. 6 to 0. 8 in 8 eyes. One eye had malignant glaucoma, and 8 eyes had cornea edema and slightly fibrin exudation in the pupil area; ln all cases maintained function conjunctival blebs of filtering, OCT confirmed this. lOP remained normal in 28 eyes in 3-6mo follow up, lOP of 2 other eyes could be controlled by anti-glaucoma eye drops.
CONCLUSlON:Double incision combined surgery of single- stab trabeculectomy and phacoemulsification is effective and safe, reduces the postoperative complications and is worthy of promotion.

A rapid, sensitive and simple liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the simultaneous determination of clevidipine butyrate and its primary metabolite clevidipine acid in dog blood. After one-step protein precipitation with methanol, the chromatographic separation was carried out on an Ecosil C18 column (150 mm x 4.6 mm, 5 µm) with a gradient mobile phase consisting of methanol and 5 mmol · L(-1) ammonium formate. A chromatographic total run time of 13.0 min was achieved. The quantitation analysis was performed using multiple reaction monitoring (MRM) at the specific ion transitions of m/z 454.1 [M-H]- --> m/z 234.1 for clevidipine butyrate, m/z 354.0 [M-H]- --> m/z 208.0 for clevidipine acid and m/z 256.1 [M-H]- --> m/z 227.1 for elofesalamide (internal standard, IS) in the negative ion mode with electrospray ionization (ESI) source. The linear calibration curves for clevidipine butyrate and clevidipine acid were obtained in the concentration ranges of 0.5-100 ng · mL and 1-200 ng · mL(-1), separately. The lower limit of quantification of clevidipine butyrate and clevidipine acid were 0.5 ng · mL(-1) and 1 ng · mL(-1). The intra and inter-assay precisions were all below 12.9%, the accuracies were all in standard ranges. Stability testing indicated that clevidipine butyrate and clevidipine acid in dog blood with the addition of denaturant methanol was stable under various processing and/or handling conditions. The validated method has been successfully applied to a pharmacokinetic study of clevidipine butyrate injection to 8 healthy Beagle dogs following intravenous infusion at a flow rate of 5 mg · h(-1) for 0.5 h.

A rapid, sensitive and simple liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the simultaneous determination of clevidipine butyrate and its primary metabolite clevidipine acid in dog blood. After one-step protein precipitation with methanol, the chromatographic separation was carried out on an Ecosil C18 column (150 mm x 4.6 mm, 5 µm) with a gradient mobile phase consisting of methanol and 5 mmol · L(-1) ammonium formate. A chromatographic total run time of 13.0 min was achieved. The quantitation analysis was performed using multiple reaction monitoring (MRM) at the specific ion transitions of m/z 454.1 [M-H]- --> m/z 234.1 for clevidipine butyrate, m/z 354.0 [M-H]- --> m/z 208.0 for clevidipine acid and m/z 256.1 [M-H]- --> m/z 227.1 for elofesalamide (internal standard, IS) in the negative ion mode with electrospray ionization (ESI) source. The linear calibration curves for clevidipine butyrate and clevidipine acid were obtained in the concentration ranges of 0.5-100 ng · mL and 1-200 ng · mL(-1), separately. The lower limit of quantification of clevidipine butyrate and clevidipine acid were 0.5 ng · mL(-1) and 1 ng · mL(-1). The intra and inter-assay precisions were all below 12.9%, the accuracies were all in standard ranges. Stability testing indicated that clevidipine butyrate and clevidipine acid in dog blood with the addition of denaturant methanol was stable under various processing and/or handling conditions. The validated method has been successfully applied to a pharmacokinetic study of clevidipine butyrate injection to 8 healthy Beagle dogs following intravenous infusion at a flow rate of 5 mg · h(-1) for 0.5 h.
Animals ; Butyrates ; blood ; pharmacokinetics ; Calibration ; Chromatography, Liquid ; Dogs ; Infusions, Intravenous ; Pyridines ; blood ; pharmacokinetics ; Tandem Mass Spectrometry

Aim TodevelopandvalidateaLC-MS/MS assay to quantify halometasone in rabbit plasma and study pharmacokinetics of halometasone after dermal topical administration of Halometasone Cream.Meth-ods Theplasmasamplewassubmittedtoliquid-liquid extraction using methyl tertiary butyl ether,with dexa-methasone as the internal standard (IS ).Chromato-graphic separations were performed on a Diamonsil C18 column(100 mm ×4. 6 mm,5 μm)with a linear gra-dient of methanol and 2 mmol · L-1 ammonium ace-tate.Halometasone and dexamethasone(IS)were ion-ized with an ESI source operated in negative ion mode, and the detected ions were m/z 503. 1→413. 0 (halo-metasone),m/z 391. 0→361. 0 (dexamethasone ). The test article could be monitored in rabbit plasma when following single dermal topical administration of Halometasone Cream at 1 g/100 cm2 to rabbits by u-singavalidatedLC-MS/MSassay.Results Calibra-tion curve was linear over the concentration range of0. 02~20 μg·L-1 in rabbit plasma.For low,medi-um,high concentration of QC solutions,the intra-and inter-day precision was in the range of 3. 72% ~7. 87%, and the accuracy was within 99. 1% to 103%. The pharmacokinetic parameters in rabbits were as follows:Tmax,Cmax,AUC0-t,T1/2 was (7. 38 ± 1. 06)h,(1. 16 ±0. 527)μg·L-1,(18. 8 ±7. 23)h·μg·L-1 ,(13. 8 ±3. 70)h,respectively.Conclusions ThisLC-MS/MSanalysismethodhashighsensitivi-ty,and sample processing method is simple,which has been rigorously validated.The method could be suc-cessfully applied to the pharmacokinetic study of halo-metasone after skin administration of Halometasone Cream to rabbits.