OBJECTIVE: To establish an HPLC-UV-ESI-MSn method for the study of impurity profile of amoxicillin and clavulanate potassium tablets. METHODS: Agilent 1100 LC/MSD Trap liquid chromatography-mass spectrometry was used, and the column was Shim pack CLC-ODS RP18(4.6 mm×250 mm, 5 μm). The mobile phase A was 20 mmol·L-1 ammonium acetate (pH adjusted to 6.0), and the mobile phase B was 20 mmol·L-1 ammonium acetate-acetonitrile (20∶80) (pH adjusted to 6.0). Gradient elution was performed at a flow rate of 1.0 mL·min-1. ESI source was used. Positive and negative ion scan was conducted with a scanning range of m/z 100-1 500. The nebulizing pressure was 275.8 kPa, dry gas flow was 9 L·min-1, and post-column diversion ratio was 1∶5. Some related substances were identified by comparing the retention time in the chromatography, [M+H]+ spectrum and MS2 spectrum with those of the reference substances, while the others which do not have reference substances were deduced or speculated by analyzing the MS2 or MSn fragmentation with the help of a rule summarized from the MS2 fragmentation of amoxicillin, clavulanic acid and system suitability impurity reference substances. RESULTS: A total of 15 related substances were separated and characterized including nine known impurities like amoxicilloic acid, amoxicillin dimer, etc. and six unknown impurities. CONCLUSION: The method can be applied in the identification and qualitative analysis of the related substances in amoxicillin and clavulanate potassium tablets and is helpful for the quality control and optimization of the synthetic process.

Objective To discuss the effect of two different warming methods on the circulation of free flaps during operation and seek for an optimal warming method which is benificial for the circulation of free flaps, and thus provide clinical evidences for intraoperative care. Methods A total of 96 patients undergoing free flap transplantation were divided into routine warming group and diversification warming group randomly (n=48 each). In routine warming group, temperature in the operating room was set at routine temperature and flaps were douched by routine heated saline. In diversification warming group, the temperature in the operation room, fluids used for washing wounds and flaps were warmed, fluids for intravenous application were pre-warmed by a fluid warmer, in addition, the operation table where patients were lying on was covered with warmer blanket. The influence of two different warming methods on the circulation of free flaps was observed. Results The scores of flap elasticity and capillary refill time in routine warming group were 2.02 ± 0.79 and 2.04 ± 0.81, respectively, vs. 2.50 ± 0.61 and 2.48 ± 0.6 in diversification warming group, showing diversification warming group was statistically better than routine warming group (Z=1.949, 3.872, P<0.05). In addition, the flap survival rate in routine warming group was 81.2%(39/48) vs. 95.8%(46/48) in diversification warming group, showing significantly better results in diversification warming group (χ2=4.02, P < 0.05). Conclusions The diversification warm keeping method can effectively promote the circulation and survival of flaps, improve clinical prognosis, accelerate postoperative rehabilitation, therefore is worthy of being recommanded its clinical application.

This paper retrospectively summarized and analyzed observation of the disease and nursing care of 17 patients with pregnancy-associated fulminant type 1 diabetes and intrauterine fetal demise.Through timely supplying blood capacity and improving renal perfusion,maintaining blood glucose homeostasis and acid-alkali,potassium balance,safety of the patients was guaranteed;by providing effective psychological nursing,puerperal dietary guidance and discharge guidance,patients' rehabilitation was improved.As a result,16 patients were in stable condition,and dead babies were delivered.Major bleeding event occurred in one patient after delivering the dead baby,and the patient developed shock as well as liver and kidney failure.The patient was transferred to ICU for further treatment and became stable and was discharged after two months.

D-myo-inositol 1,4,5-trisphosphate (IP(3)) plays an important role in signal transduction. It releases Ca(2+) from intracellular sites, which activates the Ca(2+)-dependent channels such as large-conductance Ca(2+)-activated potassium channels (BK channels). The present study was therefore designed to determine if the activity of BK channels in porcine coronary artery smooth muscle cells was increased by IP(3). Using the inside-out patch-clamp technique, the activity of single BK channels was recorded in porcine coronary artery smooth muscle cells. In excised inside-out membrane patches, IP(3) (10-50 micromol/L) enhanced the open probability (Po) of BK channels in a dose-dependent manner in the intracellular side of inside-out patches and its effect was almost completely abolished by washout. The open-state probability of the BK channels increased from a control level of 0.0402+/-0.0152 to 0.1365+/-0.0212 (20 micromol/L IP(3)) and 0.1865+/-0.0175 (30 micromol/L IP(3)). IP(3) decreased the mean close time markedly, but had no effect on the amplitude of BK channels. The activation of IP(3) on BK channels did not decline. The metabolite of IP(3) had no obvious effect on BK channels. This study provides evidence that IP(3) activates BK channels in porcine coronary artery smooth muscle cells in a dose-dependence manner.
Animals ; Coronary Vessels ; cytology ; metabolism ; Inositol 1,4,5-Trisphosphate ; physiology ; Large-Conductance Calcium-Activated Potassium Channels ; metabolism ; Muscle, Smooth, Vascular ; metabolism ; Swine ; Vasodilation ; physiology

To approach the method of isolation of tolerant human atrial myocytes, single myocytes were isolated by modified procedure of enzymatic dissociation with protease (type XXIV) and collagenase (type V). L-type calcium channel current (I(Ca-L)), sodium current (I(Na)), transient outward potassium current (I(to1)), and inward rectifier potassium current (I(K1)) in isolated atrial myocytes were recorded by using whole-cell patch-clamp techniques. Single cardiocytes isolated by this method were smooth, well-striated and rod-shaped. The yields of recordable myocytes, which viable and calcium-tolerant for electrophysiological studies, were 50%-60% of the total isolated cells. Compared with other isolation methods, this method was simple and steady, but with yield of a great number of qualified myocytes. The currents recorded in these cells were functional and active. Our research suggests that the myocytes isolated by the described method in this paper have normal electrophysiological function and are appropriate for patch-clamp experiments.
Cell Separation ; methods ; Humans ; Myocardium ; cytology ; Myocytes, Cardiac ; cytology ; Patch-Clamp Techniques

Objective To develop a method of quantitative analysis of multi-components by single marker (QAMS) for simultaneously determining five compounds in Curcumae Aromaticae Radix.Methods An HPLC method was developed as QAMS to determine curcuma diol,ocathydro-1,4-dihydroxy-1,4-dimethyl-7-(propan-2-ylidene)azulen-5(1H)-one,original curcumol and curcumin in Curcumae Aromaticae Radix,using curdione as intermal reference substance,and the relative correction factor (RCF) of the four components was determined by HPLC with good reproducibility.Their contents in 10 batches of samples,collected from different areas,were determined by both external standard method and QAMS.Result No significant differences were found in the quantitative results of four compounds in 10 batches of Curcumae Aromaticae Radix determined by external standard method and QAMS.Conclusion It is feasible and suitable to evaluate the quality of Curcumae Aromaticae Radix by QAMS.

AIM To establish an HPLC method for the simultaneous content determination of four constituents in Liujing Toutong Tablets (Angelicae dahuricae Radix,Magnoliae Flos,Ligustici Rhizoma et Radix,etc.).METHODS The analysis of 30％ ethanol extract of this drug was performed on a 35 ℃ thermostatic Waters C18 column (4.6 mm × 250 mm,5 μm),with the mobile phase comprising of methanol-4％ acetic acid flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelength was set at 320 nm.RESULTS Puerarin,ferulic acid,imperatorin and isoimperatorin showed good linear relationships within the ranges of 60.6-303 μg/mL (r=0.999 9),1.59-7.95 μg/mL (r =0.999 9),1.57-7.85 μg/mL (r =0.999 9) and 0.752 5-3.762 5 μg/mL (r =0.999 7),whose average recoveries (RSDs) were 97.75％ (1.7％),97.68％ (2.3％),97.94％ (1.0％) and 98.29％ (1.6％),respectively.CONCLUSION This stable and reliable method can be used for the quality control of Liujing Toutong Tablets.

This study aimed to determine and quantitate the mammographic and sonographic characteristics in 13 cases of solid neuroendocrine breast carcinoma (NEBC) and to analyze the association of radiological findings with the clinical and histopathologic findings. The clinical data and imaging findings of 13 female patients with histologically confirmed solid NEBC were reviewed. Imaging data were evaluated by two radiologists for a consensual diagnosis. All patients presented with one palpable mass; only 1 experienced occasional breast pain, and 5 complained of fluid discharge. In 7 patients, the masses were firm and mobile. Regional lymph node metastasis was noted in only 1 patient. For the 10 patients who underwent mammography, 6 had a mass, 1 had clustered small nodules with clustered punctuate microcalcifications, 2 had asymmetric focal density, and 1 had solitary punctuate calcification. Most of the masses had irregular shape with indistinct or microlobulated margins. For the 9 patients who underwent ultrasonography (US), 9 masses were depicted, all of which were hypoechoic, mostly with irregular shape and without acoustic phenomena. Different types of acoustic phenomena were also identified. One patient had developed distant metastases during follow-up. NEBC has a variety of presentations, but it is mostly observed on mammograms as a dense, irregular mass with indistinct or microlobulated margins. Sonographically, it typically presents as an irregular, heterogeneously hypoechoic mass with normal sound transmission. Histories of nipple discharge and calcification observed using imaging are not rare.
Adult ; Aged ; Biopsy, Fine-Needle ; Breast Neoplasms ; diagnostic imaging ; metabolism ; pathology ; Calcinosis ; diagnostic imaging ; Carcinoma, Neuroendocrine ; diagnostic imaging ; metabolism ; pathology ; Chromogranin A ; metabolism ; Female ; Follow-Up Studies ; Humans ; Ki-67 Antigen ; metabolism ; Lymphatic Metastasis ; Mammography ; Middle Aged ; Phosphopyruvate Hydratase ; metabolism ; Receptors, Estrogen ; metabolism ; Receptors, Progesterone ; metabolism ; Synaptophysin ; metabolism ; Ultrasonography, Mammary

OBJECTIVETo explore the possible role of CD28/CTLA-4 co-stimulators in immune pathophysiology of acquired aplastic anemia(AA).

METHODSBy FACS, the percentages of CD28, CTLA-4 expressing CD3+ CD4+ T cells and the level of Th1, Th2 in bone marrow were detected in 23 AA patients at active phase, 10 at recovery phase and 15 normal controls. The relationship between the co-stimulators, Th1, Th2, and absolute neutrophil counts (ANC) was evaluated.

RESULTS(1) The percentage of CD28 and CTLA-4 expressing CD3+ CD4+ T cells in bone marrow, and CD28+/CTLA-4+ ratios were (31.40 +/- 10.83)%, (2.45 +/- 1.30)% , and 17.02 +/- 13.44 in normal controls respectively, (39.84 +/- 10.89)%, (1.43 +/- 0.67)%, and 43.04 +/- 37.61 in AA at active phase, respectively, (22 +/- 9.08)%, (3.46 +/- 2.26)%, and 10.49 +/- 7.8 in AA at recovery phase, respectively. The percentage of CD28 and CD28+/CTLA-4+ ratio were significantly higher, while CTLA-4 were lower in active phase AA patients than in normal controls (P < 0.05). These values in recovery phase AA were comparable to those in normal controls. (The Th1, Th2, and Th1/Th2 in bone marrow were (4.21 +/- 2.11)%, (1.99 +/- 1.27)%, and 2.46 +/- 1.28 in normal controls respectively, (11.13 +/- 4. 96)%, (2.46 +/- 1.65)%, and 5.20 +/- 1.98 in active phase AA and (5.39 +/- 4.2)9%, (2.53 +/- 2.41)%, and 2.87 +/- 1.43 in recovery phase AA, respectively. The percentage of Th1 and Th1/Th2 ratio were significantly higher in AA patients at active phase than in normal controls (P < 0.05). (3) The CD28+/CTLA-4+ ratio was positively related to the Th1+ /Th2+ ratio (P < 0.05). ANC was negatively related to CD3+ CD4+ CD28+ T cells (P < 0.01), and positively to CD3 + CD4 ' CTLA-4' T cells (P < 0.01) respectively.

CONCLUSION(1) The expression of CD28 was increased while CTLA-4 decreased on the membranes of CD3+ CD4+ T cells in bone marrow of AA patients. (2) The abnormal expression of CD28 costimulator promoted the shift of immune balance to Thl type. (3) The unbalance of CD28+ / CTLA-4+ is important for the immune pathophysiology of AA.

Adolescent ; Adult ; Aged ; Anemia, Aplastic ; immunology ; metabolism ; Antigens, CD ; immunology ; metabolism ; CD28 Antigens ; immunology ; metabolism ; CD4-Positive T-Lymphocytes ; metabolism ; CTLA-4 Antigen ; Child ; Female ; Humans ; Male ; Middle Aged ; Th1 Cells ; immunology ; Th2 Cells ; immunology

OBJECTIVETo explore the scan technique and image quality of coronary angiography with dual source computed tomography (CT) without oral metoprolol preparation.

METHODSPlain and enhanced dual source CT coronary angiography without oral metoprolol preparation was prospectively performed in 600 patients. Calcium scoring with plain scan images as well as multi-planar reconstruction (MPR), maximum intensity projection (MIP), and volume rendering technique (VRT) reconstruction with enhanced scan images were performed in all cases. The scan technique and post-reconstruction experience was summarized. The image quality was classified as 1 to 4 points, and coronary segments classified according to the American Heart Association standards were evaluated.

RESULTSThe average calcium score of the 600 cases was 213.6 +/- 298.7 (0-3,216.5). The average heart rate of the enhanced scan was 82.1 +/- 16.2 (47-139) bpm. The post-reconstruction methods with which coronary segments could be shown as best as possible consisted of single phase reconstruction method, two or more phases supplemented method, and electrocardiogram editing method. Altogether 8,457 coronary segments were evaluated, among which 97.2% were evaluated as point 1, 1.7% point 2, 0.5% point 3, and 0.6% point 4. The coronary segments in 261 cases were completely normal, while 360 segments were diagnosed with < 50% stenosis and 625 segments with > or = 50% stenosis.

CONCLUSIONSExcellent coronary artery image can be obtained with dual source CT in patients with any heart rate without oral metoprolol preparation. Heart rate is not a major source of the artifact, coronary segments can be well shown with single or multiple-phase reconstruction method.

Adult ; Coronary Angiography ; Female ; Heart Rate ; Humans ; Male ; Middle Aged ; Tomography, X-Ray Computed ; methods