Objective To develop a high performance liquid chromatography-ultraviolet detection(HPLC-UV) method for the determination of midazolam in rat plasma,and to study the pharmacokinetics of midazolam in single dose intravascular administration in rats.These results can provide a methodological reference for evaluating cytochrome P450 3A(CYP3A) activity.Methods Firstly,common carotid artery(CCA) intubate was set in rats,Midazolam was injected into rat vena caudalis and plasma samples were collected in different time.Then,the sample was extracted using dichlormethane and evaporated to dry thoroughly with N2 soft stream at 37 ℃.Finally,the residues were reconstituted with 150 ?L 30 %methanol and further analyzed by HPLC.A Phenomenex C18 column(4.6?150 mm i.d,5 ?m)was employed at 20 ℃.The mobile phase consisted of(A) methanol-acetonitrile-0.03 %phosphate solution(pH 2.85) in the proportion of 5:10:85(V/V/V) and(B) methanol-acetonitrile-0.03 %phosphate solution(pH 2.85) in the proportion of 5:10:85(V/V/V),using a linear gradient elution of 0 %B～100 %B at 0～18 minutes,then retaining for 5 minutes and returning to A.The flow rate was set at 0.5 mL/min and the ultraviolet detector was operated at 240 nm.Results Midazolam and internal standard were isolated on baseline in plasma apart from all other material.The the linear range was from 0.025～2.0 ?g/mL,and the detection limit was 2.5 ng/mL.The standard addition recoveries were from 99.98 %to 105.71 %and the extraction rates were from 91.29 %to 92.58 %.All of the intraday and interday variations were less then 14 %.The primary pharmacokinetics parameters of rat single dose intravascular administration were as follows:t1/2?was 0.582 h,Vd was 0.214 L,Cl was 0.584 L/h and AUC0→t was 0.419 ?g?h-1?mL-1.Conclusion Midazolam single dose intravascular administration has the characteristics of rapid distribution and elimination in blood and quick transport from blood to tissue.The method is sensitive,simple and suitable for the research of pharmacokinetie parameters of midazolam and description of possible pharmacological interactions of rat CYP3A1/2 or human CYP3A4/5 enzymes.

Objective To observe the effect of Zhipu Hollow Suppository (ZHS) on postoperative ankylenteron in rats,and to investigate the possible mechanism.Methods Forty Wistar rats were randomly divided into model group,ZPS group,positive control group,and blank control group,10 rats in each group.Rats were given corresponding drugs according to the experimental design after modeling.Rats were executed after medication for one week,and the pathologic changes of ileal serosa were observed under scanning electron microscope and the ultrastructure of ileal serosa cells was observed under transmission electron microscope.Results Results of the examination under scanning electron microscope showed that the arrangement of ileal-serosa mesothelial cells in ZHS group was regular,and the network structure arranged spindle-shaped with uniformity size and with the shape close to the normal ones.There was proliferation in mesothelial cells in the exposed base layer.Under the transmission electron microscope,mesothelial cells recovered well and the proliferation of fibroblasts was not obviously in ZHS group,but mesothelial cells disappeared and the secretion of collagen by fibroblasts was actively in the model group.Conclusion ZHS can prevent ankylenteron by promoting the ileal-serosa mesothelial cell proliferation,improving the ileal-serosa ultrastructure and protecting the ileal serosa.

OBJECTIVE: To establish a simple and rapid HPLC method for the determination of serum concentration of isofosfamide.METHODS: With cyclophosphamide as internal standard,the serum sample was extracted on C18 solid phase extraction column and determined on Zorbox-ODS C18 at room temperature.The mobile phase consisted of acetonitrile-water(25∶75) at a flow rate of 1 mL?min-1 and a detection wavelength of 200 nm.RESULTS: The linear range of ifosfamide was 2.5～200 ?g?mL-1(r=0.999 9),with the lowest detectable limit at 0.5 ?g?mL-1(S/N=3).Both the intraday and interday RSD were less than 5%.The extraction recoveries of ifosfamide at high,middle and low concentrations were ranged between 89.3% and 93.6% versus 77.6%～82.3% in chloroform-treated serum samples.CONCLUSION: The method which uses the C18 solid phase extraction column is of high recovery,and the sample treatment by which is simple,rapid,accurate,and pollution-free,and it is applicable for the monitoring of serum concentration and pharmacokinetic study of isofosfamide.

Objective To establish a new kinetic spectrophotometry for determination of formaldehyde in water sample. Methods Micro formaldehyde could sensitively catalyze the discoloring reaction of methyl orange oxidized by potassium bromated in dilute H3PO4 medium, the relationship between the reaction speed and the content of formaldehyde could be determinated by spectrophotometry, then the content of formaldehyde could be determinated. Results The optimum condition and kinetic paramneters were investigated and showed in the present paper in detail. The calibration curve showed linearity in the range of 0.13-3.0 ?g/ml and the limit of detection was 4.7?10-5 mg/ml. The RSDs ranged from 1.5% to 3.3%.The recovery rates ranged from 97.3% to 102.0%. Conclusion The mothed was simple, rapid and more sensitive, can be used for the determination of micro amounts of formaldehyde in water sample with satisfied results.

[Objective] To compare the antipyretic and anti-inflammatory actions and acute toxicity between Chuanxinlian Soft Capsules (CSC) and Chuanxinlian Tablets. [Methods] Antipyretic action of CSC was observed on rabbit fever models induced by lipopolysaccharide (LPS) and rat fever models induced by baker's yeast. Rat models with pedal swelling induced by carrageenin, mouse models with auricle swelling induced by dimethylbenzene and mouse models with celiac capillary hyperpermeability induced by acetic acid were used to observe the anti-inflammatory action oi CSC. Its acute toxicity in mice was also evaluated. [Results] High- and moderate-dose CSC reduced fever in rabbits induced by LPS (P 0.05). High-, moderate- and low-dose CSC has an inhibitory effect on acute inflammatory exudation (P

BACKGROUND:Conventional surgical repair can cause large traumas in patients with knee injuries, and patients often recover slowly after implant fixation, most of whom can appear to have poor recovery of knee function. OBJECTIVE: To explore the folow-up effect of arthroscopic double-steel wire clip fixation on tibial eminence avulsion fracture of anterior cruciate ligament. METHODS: A retrospective analysis was performed on the clinical data of 23 patients with tibial eminence avulsion fractures, who were given arthroscopic double-steel wire clip fixation. The patients were folowed up for 1-6 months. Short- and middle-term therapeutic effect as wel as IKDC and Lysholm scores before and after treatment were observed and analyzed. RESULTS AND CONCLUSION:The operation time was 35-65 minutes, and no complications, such as blood, nerve and anterior cruciate ligament injuries occurred. Moreover, no infection and other poor biocompatible reactions occurred after internation fixation. Al patients were folowed up for 1-6 months. The excelent and good rate was 87% at 1 month after treatment and 96% at 6 months after treatment. Al the patients had improved IKDC score and Lysholm score after treatment (P < 0.05), indicating that the knee function of patients was improved significantly.

Objective To explore the optimal method of extraction and determination of rhynchophylline and isorhynchophylline in Ramulus Uncariae cum Uncis. Methods A RP-HPLC was performed on a Phenomenex C18 column (4. 6 mm× 150 mm, 5m) at 30℃. The mobile phase consisted of methanol-0. 01 mmol/L triethylamine solution (70: 30, pH 7.0 adjusted by glacial acetic acid) at the flow rate of 0. 5 mL/min. The automatic sample injector was set at 5℃ and the ultraviolet detector was operated at 245 nm. And then, the effect of different extraction condition on the contents of rhynchophylline and isorhyn-ehophylline in Ramulus Uncariae cum Uncis was investigated. ResultsA good linearity of rhynchophylline was in the range of 2. 5μg/mL to 80. 0μg/mL (Y=76. 7170X-0. 0727,r=0. 999 8),the average recoveries were from 99. 84 ％ to 116. 91％, and the RSD (n=6) were less than 8.8 ％. A good linearity of isorhynchophylline was in the range of 2. 0 μg/mL to 80. 0 μg/mL (Y=87. 4729X-0. 3666, r=0.999 7), the average recoveries were from 87.08 ％ to 104. 97 ％, and the RSD (n=6) were less than 7.3 ％. The contents of rhynchophylline and isorhynchophylline in the methanol-extracted solu- tion were approximately ten times as much as those in water-decocted solution. The main factors which had effect on the ex-traction of rhynchophyUine and isorhynchophylline in methanol solution were supersonic time,methanol amount and cold-dousing time, in a decreasing sequence. The best condition selected by orthogonal experiment wasb as follows: extracting the medicinal material with 20-fold volumes of methanol, cold maceration for 24h and supersonic extraction for 1 h. Conclusion The extraction percentage of rhynchophylline and isorhynchophylline in the extraction condition of cold maceration with methanol and supersonic extraction is superior to that in the water-decocting condition. The method is simple, fast and accurate, and it can be used for the quality control of Ramulus Uncariae cum Uncis.

Fast progresses in stem cell-based tooth tissue engineering have been achieved in recent years in several animal models including the mouse, rat, dog, and pig. Moreover, various postnatal mesenchymal stem cells of dental origin have been isolated and shown capable of differentiating into odontoblasts and generating dentin. Meanwhile, human keratinocyte stem/progenitor cells, gingival epithelial cells, and even iPSC-derived epithelium have been demonstrated to be able to differentiate into functional ameloblasts. Translational medicine studies in the nonhuman primate are irreplaceable steps towards clinical application of stem cell-based tissue engineering therapy. In the present study, we first examined the epithelial stem cell markers in the rhesus skin using immunostaining. Keratinocyte stem cells were then isolated from rhesus epidermis, cultured in vitro, and characterized by epithelial stem cell markers. Epithelial sheets of these cultured keratinocytes, which were recombined with E13.5 mouse dental mesenchyme that possesses odontogenic potential in the presence of exogenous FGF8, were induced to differentiate into enamel-secreting ameloblasts. Our results demonstrate that in the presence of appropriate odontogenic signals, rhesus keratinocytes can be induced to gain odontogenic competence and are capable of participating in odontogenesis, indicating that rhesus keratinocytes are an ideal epithelial cell source for further translational medicine study of tooth tissue engineering in nonhuman primates.
Ameloblasts* ; Animals ; Dentin ; Dogs ; Epidermis ; Epithelial Cells ; Epithelium ; Humans ; In Vitro Techniques ; Keratinocytes* ; Macaca mulatta ; Mental Competency ; Mesenchymal Stromal Cells ; Mesoderm* ; Mice* ; Models, Animal ; Odontoblasts ; Odontogenesis ; Primates ; Rats ; Skin ; Stem Cells ; Tissue Engineering ; Tooth ; Translational Medical Research