A rapid and sensitive liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS) method for quantification of sitagliptin in human plasma and urine had been developed. This method was applied to the pharmacokinetics study of sitagliptin tablet after single- and multiple-dosing in Chinese population. Plasma samples were prepared by a liquid-liquid extracted method, and urine samples were diluted. Compounds were analyzed by multiple reaction monitoring (MRM) mode with a electrospray ionization (ESI) interface. Mobile phase consisted of methanol and water (85 : 15, v/v). The linear concentration range of calibration curve was 0.5-1 000 ng.mL-1. and 0.2-100 µg.mL , intra-run/between-run accuracy was 98.98%-103.69% and 97.63%-102.29%, intra-run/between-run precision was <5.51% and 4.26% for plasma and urine sample, respectively. The stability of sitagliptin stock solution was tested for 55 days at -30 °C. Sitagliptin was stable when stored under the following conditions: 24 hours in the autosampler after sample preparation; 24 hours at room temperature, after 3 freeze and thaw cycles (from -30 °C to room temperature), 40 days at -30 °C for plasma and urine samples. The absolute recovery in plasma was 71.1%, and no matrix effect was founded. This method was proved simple, specific, sensitive, rapid and suitable for pharmacokinetics study of sitagliptin in human being.
Calibration ; Chromatography, Liquid ; Humans ; Liquid-Liquid Extraction ; Sitagliptin Phosphate ; blood ; pharmacokinetics ; urine ; Tandem Mass Spectrometry

Objective: To study the protective effects and the mechanism of Salvia yunnanensis extract on hypoxia/reoxygenation injury in cultured rat H9c2 cardiomyocytes.Methods: The hypoxia/reoxygen (H/R) injury model was established in H9c2 cell strain with or without the extract of Salvia yunnanensis.The cultured H9c2 cardiomyocytes were randomly divided into 6 groups: the normal control (C) group, H/R group, H/R+verapamil (H/R+V) group, H/R+Salvia yunnanensis extract at low dose (H/R+L, 0.01 mg·L-1) group, medium dose (H/R+M, 0.1 mg·L-1) group and high dose (H/R+H, 1.0 mg·L-1) group.The cell viability was measured by MTT assay and the activity of malondialdehyde (MDA) and lactate dehydrogenase (LDH) was measured by a detection kit.Fluorescence absorbance (A) value was measured by a fluoroscopy to show the intracellular reactive oxygen species (ROS) levels.Results: Compared with that in the model group, the survival rate of myocardial cells was significantly higher in Salvia yunnanensis extract at low, medium and high dose groups (P＜0.05 or P＜0.01), and the intracellular LDH leakage (P＜0.05 or P＜0.01), the content of MDA in cytoplasm (P＜0.01) and the intracellular ROS levels significantly decreased in Salvia yunnanensis extract at high dose group (P＜0.05).Conclusion: The extract of Salvia yunnanensis has protective effect on hypoxia/reoxygenation injury in cultured rat H9c2 cardiomyocytes, and the mechanism may be related to the reduction of lipid peroxides and removal of cell oxygen free radicals.

A rapid and sensitive liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS) method for quantification of sitagliptin in human plasma and urine had been developed. This method was applied to the pharmacokinetics study of sitagliptin tablet after single- and multiple-dosing in Chinese population. Plasma samples were prepared by a liquid-liquid extracted method, and urine samples were diluted. Compounds were analyzed by multiple reaction monitoring (MRM) mode with a electrospray ionization (ESI) interface. Mobile phase consisted of methanol and water (85 : 15, v/v). The linear concentration range of calibration curve was 0.5-1 000 ng.mL-1. and 0.2-100 µg.mL , intra-run/between-run accuracy was 98.98%-103.69% and 97.63%-102.29%, intra-run/between-run precision was <5.51% and 4.26% for plasma and urine sample, respectively. The stability of sitagliptin stock solution was tested for 55 days at -30 °C. Sitagliptin was stable when stored under the following conditions: 24 hours in the autosampler after sample preparation; 24 hours at room temperature, after 3 freeze and thaw cycles (from -30 °C to room temperature), 40 days at -30 °C for plasma and urine samples. The absolute recovery in plasma was 71.1%, and no matrix effect was founded. This method was proved simple, specific, sensitive, rapid and suitable for pharmacokinetics study of sitagliptin in human being.

Objective To clear the clinical efficacy of using early rehabilitation nursing model in the course of clinical nursing in patients with cerebral infarction.Methods Divided 60 patients with cerebral infarction into the observation group (30 cases) and the control group (30 cases) randomly,traditional nursing measures were used in the control group,the early rehabilitation nursing model was used in the observation group in addition.Compared the clinical efficacy,ADL scores,NIHSS scores and the satisfaction between the two groups.Results In the observation group,the satisfaction was 93.33％ (28/30),the efficiency was 90.00％(27/30),which were significantly higher than those in the control group [76.67％(23/30),[66.67％(20/30)].The ADL and NIHSS scores in the observation group was (91.25±11.38) and (8.01±1.05) respectively,which were significantly different with those of in the control group,the ADL and NIHSS scores was (79.37±10.27) and (10.22±1.27) respectively in the control group.Conclusions Early rehabilitation nursing model can effectively improve the never function of patients with cerebral infarction,and then improve their quality of life.

BACKGROUND:Recovery of motor and sensory function from peripheral nerve injury is relatively slow and incomplete. It is a difficult problem for orthopedic surgeons that mainly leads to the decline in the quality of life in patients.
OBJECTIVE: To conclude the methods and corresponding outcomes in peripheral nerve regeneration by analyzing the new treatment means for peripheral nerve injury.
METHODS:PubMed, Wanfang, CNKI databases were retrieved for relevant articles using key words of “nerve injury, regeneration”, and then retrieval data were sorted and analyzed.
RESULTS AND CONCLUSION:In recent years, in-depth studies on peripheral nerve repair have been made in the folowing aspects: surgical mode, drug, cytokine, gene transfer and biomaterials as wel as traditional Chinese medicine. If the detect size is four times longer than the diameter of nerves, the nerve regeneration chamber can achieve good outcomes. The methods of restoring nerve continuity folowing nerve injury are developed from surgical anastomosis to photochemohistological method, thermal laser welding, plastic repair and other emerging technologies. Studies have found that plasminogen activator, nerve growth factor, neurotrophic factor, recombinant erythropoietin, human tissue kalikrein, B vitamins and their derivatives, herbal preparations, immunosuppressive agents al can promote nerve regeneration.

BACKGROUND:Studies have shown that craniocerebral injury can promote the repair of sciatic nerve injury in rats, but its precise mechanism remains unclear.
OBJECTIVE:To further explore the action mechanism of craniocerebral injury on the repair of sciatic nerve injury using morphology and histology.
METHODS:Sixty specific-pathogen-free healthy male Sprague-Dawley rats were randomly divided into two groups. Rats with craniocerebral injury and sciatic nerve injury were considered as the experimental group. Rats with simple sciatic nerve injury were considered as the control group. Classical Feeney method was used in models of craniocerebral injury and SunderlandV sciatic nerve injury. At 8 and 12 weeks after modeling, sciatic nerve index was detected. Masson staining and NF200 immunofluorescence staining were used to observethe nerve regeneration atthe anstomotic site. Transmission electron microscope was used to observe the number of regenerative axons.
RESULTS AND CONCLUSION:At 8 and 12 weeks after modeling, compared with the control group, gait and sciatic nerve index recovered better in the experimental group. In the experimental group, Masson staining showed fewer nerve membrane colagen fibers, and the axon arranged neatly.NF200 immunohistochemistry showed that in the experimental group, the density of regenerated nerves was high, and nerveswere regularly distributed. Transmission electron microscopy showed that in the experimental group, regenerative axons were regularly arranged, colagen scar was less, and myelin layer arranged regularly. Results suggested that the craniocerebral injury in rats may promote the repair of peripheral nerve injury by reducing scar colagen in nerve endings.

Objective To observe the effect of total intravenous anesthesia (TIVA) on intrapulmonary shunt fraction and arterial oxygenation during one-lung ventilation (OLV) for thoracoscope surgery.Methods Forty patients scheduled for thoracoscope surgery were randomly assigned to two groups ( n =20),group of TIVA (A) and group of intravenous anesthesia combined with inhalational anesthesia(B).After inducing and intubating,patients were assigned to maintenance of anesthesia with propofol ( group A)or with sevoflurane ( group B) in order to maintain a BIS between 40 and 60.Mean arterial pressure (MAP),heart rate (HR),SpO2 and Paw were measured in four phases,always in the lateral position,10min after beginning two-lung ventilation (TLV),15 min after beginning OLV (OLV + 15 ),30 rain after beginning OLV ( OLV + 30) and 60 min after beginning OLV ( OLV + 60).Blood samples were drawn simultaneously and analyzed within 5 min.The Qs/Qt at each phase was calculated.Adverse events including hypotension,bradycardia,hypoxemia,delayed emergence and restlessness in recovery period were recorded.Results In all patients,a decrease in PaO2 and an increase in the Qs/Qt occurred during OLV were observed.But PaO2 values in group A were significantly higher than those in group B ( 177 ±88 vs 125 ±63;150 ±65 vs 110 ±67;188 ±69 vs 128 ±52) ( P ＜0.05).The Qs/Qt in group B was significantly higher than those in group A (34.2 ±5 vs 28.8 ±2;38.4 ±8 vs 32.1 ±6;37.1 ±2 vs 29.5 ±2,P ＜0.05).MAP values in group A were significantly lower than those in group B at the phase:OLV + 15 and OLV +30(72 ± 10 vs 88 ± 14;74 ± 12 vs 89 ± 10) ( P ＜ 0.05 ).The incidence of hypotension and delayed emergence in group A was higher than those in group B ( 10 case vs 4 case;9 case vs 2 case).The incidence of restlessness in recovery period in group B was more than those in group A (9 case vs 3 case).The differences between two groups were significant ( P ＜ 0.05).Conclusions Compared with sevoflurane-sufentanyl combined anesthesia,TIVA with propofol can efficiently decrease intrapulmonary shunt fraction and improve arterial oxygenation during OLV for thoracoscope surgery,which is good for the prevention of hypoxemia.

Objective Through the cell block technique to detect the expression of P16 protein in the liquid-based cytology with atypical squamous cells of undetermined significance (ASCUS) and high degree of cervical intraepithelial lesions (HSIL),to explore the significance of P16 protein in ASCUS re-evaluate.Methods Collected in our hospital in 2012 cervix liquid based cytology specimens of 45 patients,including of 15 ASCUS,11 HSIL cases,low in 11 cases of epithelial lesions (LSIL) and 2 cases of squamous cell carcinoma,2 cases of atypical glandular cells,4 cases of normal cells as a control.Immunocytochemical analysis of P16 protein control analysis,cytology and histology results.Results The expressing of P16 protein in normal cells,ASCUS,LSIL,HSIL,squamous cell carcinoma,atypical glandular cells in the positive expression rates were 0,20％,27.2％,63.6％,100％,100％.Cytology and biopsy results,cytologic diagnosis of ASCUS 15 cases,biopsy:12 cases of cervicitis,CIN Ⅱ-Ⅲ in 3 cases; cytology the in LSIL11,biopsy:5 cases of cervicitis,CIN Ⅰ 6 cases ; the cytological diagnosis HSIL11 cases,biopsy:cervical four cases of intlammation,CIN Ⅱ-Ⅲ ; cytologic diagnosis of atypical glandular cells in 2 cases,biopsy:adenocarcinoma; cytologic diagnosis of squamous cell carcinoma in 2 cases,biopsy:squamous cell carcinoma.Conclusion Detection of P16 protein on the cell block can be used for ASCUS classification ASCUS reassessment.

Objective To evaluate the uncertainty of the pseudo-ginsenoside GQ (PGQ) concentration in human plasma by HPLC-MS/MS.Methods The whole process of PGQ determination by HPLC-MS/MS in human plasma was evaluated and the uncertainty caused by repeatability,weighing,standard solution preparation,biological sample preparation,extraction recovery process,recovery,instrument precision and calibration curve fitting were evaluated,respectively.The combined and expanded uncertainty values were both calculated.Results The expanded uncertainty values for low (15.16 ng·mL-1),medium (2 516.67 ng·mL-1) and high (3 902.00 ng·mL-1) levels of PGQ were 1.39,177.74 and 262.69 ng·mL-1,respectively (P =95 ％,k =2).Conclusion The uncertainty of the PGQ determination in human plasma by HPLC-MS/MS is mainly caused by recovery,repeatabihty and sample preparation at low concentration,by sample preparation and recovery at medium and high concentration.

Diagnosis, Differential ; Humans ; Infant ; Male ; Neuroectodermal Tumors, Primitive ; pathology ; Orbital Neoplasms ; pathology