Objective:To establish the qualitative and quantitative methods for Corni Fructus in Guishao Dihuang pills. Methods:A TLC method was used to identify morroniside and loganin. An HPLC method was used to determine the content of morroniside and lo-ganin in Corni Fructus. Results:The TLC spots were clear with good separation and specificity. The linear range of morroniside was within 0. 010-2. 620 μg (r=0. 999 9) and the average recovery was 102. 75% (RSD=1. 40%, n=6). The linear range of loganin was within 0.009-2.170 μg (r =0.999 9), and the average recovery was 103.59% (RSD =1.10%, n =6). Conclusion: The method is accurate, simple and feasible with good repeatability, which can be used for the quality control of Corni Fructus in the prepa-ration.

Objective To investigate the role and potential mechanism of interleukin-17A (IL-17A) in the inflammatory response to traumatic brain injury (TBI) in rats.Methods The adult male Wistar rats were randomly (random number) divided into seven groups:control group (n =6),sham operation group (n =6),TBI group (n =24),sham operation + normal saline group (n =6),sham operation + Y320 (an immunomodulator acts as an inhibitor of IL-17A) group (n =6),TBI + normal saline group (n =6) and TBI + Y320 group (n =6).The TBI model of rat was established by using free-falling-body impact device.The levels of IL-17A and nuclear transcription factor kappa B p65 (NF-κB p65) in the cerebral cortex were assayed by using Western Blot.The capability of leaming and memory of rats was assessed by Morris water maze.The beam balance test was employed to evaluating the neurological motor performance and the capability of balance.Results Compared with the sham operation group,the levels of IL-17A and NF-κB p65 in the cerebral cortex of TBI,TBI + saline and TBI + Y320 groups increased significantly (P ＜0.05) and peaked at the 3rd day after TBI.Compared with TBI + normal saline group,the level of NF-κB p65 was significantly down regulated by Y-320 (P ＜ 0.05) at the 3rd day after TBI in TBI + Y320 group.The lengths of latency time required for rats to escape to the platform area in TBI + normal saline group were (57.72±3.29) s,(55.63±3.85) s,and (55.02±3.92) sat the3rd,5th and7th days after TBI,respectively;while those in TBI + Y320 group were (35.45 ± 3.04) s,(30.98 ± 2.92) s,and (23.90 ±2.51) s at the 3rd,5th and 7th days after TBI,respectively.Thus,the capability of learning and memory of rats in TBI + Y320 group was improved significantly 3d,5d and 7 days after TBI (all P ＜ 0.01).Conclusions This study shows IL-17A is involved in the process of secondary brain injury after TBI,and associated with inflammation by activating the NF-κB p65 signaling pathway.

Objective To compare the effect of injection with botulinum toxin type A (BTX-A) before serial casting with BTX-A injection alone for the treatment of spastic equinus in children with cerebral palsy (CP).Methods Sixty patients were divided into an experimental group and a control group with 30 patients in each.Those io the experimental group received a BTX-A injection followed by serial casting,while the controls received BTX-A only.Before treatment and 1 month,3 months and 6 months after treatment,the dorsiflexion range of motion (ROM)of the ankle recorded while the knee in flexion and extension were measured,and gait was evaluated with an observational gait scale.Results Before treatment there was no significant inter-group difference in any of the 3 outcome measures.At 1 montb,3 months and 6 months after treatment,there were significant inter-group differences in all 3 measures,and also significant differences compared with before treatment in both groups.Conclusion Lower muscle tone,greater ankle mobility and better gait patterns can be promoted in CP children with spastic equinus using serial casting combined with BTX-A injection.The improvements may last longer than those after BTX-A injection alone.

Objective To establish VX2 hepatic carcinoma model in rabbits by implanting the tumor fragment into the liver through percutaneous puncture under ultrasound guidance and to observe its growing and metastatic characteristics, to determine the optimal time for interventional experiment study with the model. Methods Inoculation of VX2 carcinoma fragment was performed in 28 New Zealand white rabbits. PET/CT and ultrasonography (US) examinations were carried out in the second, third and forth week after the inoculation, and each time two tumor-bearing rabbits were sacrificed for pathologic study. Results The successful rate of model establishment was 89.28% (25/28). On PET or CT scans, single lesion in the liver was demonstrated in 25 rabbits. Two, three and four weeks after the inoculation, the maximum diameter of the tumor was (4.82±0.80) mm, (16.05±2.89) mm and (30.08±5.38) mm respectively, while the metastasis rates was 0% (0/25), 13.04% (3/23), 76.19% (16/21) respectively. No significant necrosis was found in the second week after inoculation, only tiny coagulation necrosis was revealed in the third week, and massive necrosis was seen in the forth week. Conclusion Percutaneous inoculation of the tumor fragment into the liver under ultrasonographic guidance is a simple method to establish VX2 hepatic carcinoma in rabbits with a high successful rate. The third week after inoculation is the suitable time for making interventional experiment study.

Animals ; Apolipoproteins E ; deficiency ; genetics ; Blood Vessels ; anatomy & histology ; Fluorescence ; Lipofuscin ; Mice ; Mice, Knockout

OBJECTIVETo explore the effects of inhaled corticosteroids (ICS) on the distribution of throat flora in children with bronchial asthma.

METHODSSixty healthy children were included in the study as the control group and 160 children with asthma in acute period before ICS therapy were chosen as the experimental group. The experimental group were treated with ICS therapy. In this group, 89 children were followed up for 3 months, 68 for 6 months and 60 for 12 months. The ICS in the study was budesonide with the trade name as Pulmicort. Swab from the pharynx was used, then inoculated in agar plate. The bacteria were isolated, the distribution and variation of the microbial population in pharyngeal portion were evaluated. The data collected were analyzed using SPSS12.0 software.

RESULTSBacteria could be detected in all samples collected from children with asthma in acute, untreated period, which were mainly non-β-hemolytic streptococcus and gram negative cocci bacteria. Gram negative bacilli, streptococcus pneumoniae and mycetes were less. There were no significant differences (χ(2) value were 4.7441, 7.8582 and 1.5583 respectively, Fisher exact value were 0.0699, 0.6398, 0.2433, 0.8580, 0.6616, 0.6339, and 0.8479 respectively, P > 0.05) among children with asthma in acute period, children with asthma treated with ICS after 3, 6 and 12 months and control group. Three strains of mycetes were detected in the experimental group, and one strain in children with asthma treated with ICS for 6 months group.

CONCLUSIONSThere were no significant changes in the distribution of throat flora between the control group and the experimental group. The throat microbiology did not show significant change. Inhaled corticosteroids had no obvious effect in throat flora in children with asthma after being used for a short term and for 12 months, which suggested that inhaled corticosteroids was safe in bronchial asthma treatment. Dynamic monitoring of throat flora while the inhaled corticosteroids are used is of clinical significance.

Administration, Inhalation ; Adolescent ; Adrenal Cortex Hormones ; administration & dosage ; therapeutic use ; Asthma ; drug therapy ; microbiology ; Bacteria ; isolation & purification ; Case-Control Studies ; Child ; Child, Preschool ; Female ; Fungi ; isolation & purification ; Humans ; Male ; Pharynx ; microbiology

Objective: To investigate the antihypertensive time-effect and dose-effect features of Sancao jiangya decoction(SCD).Methods: The blood pressure of spontaneously hypertensive rats at different time points were measured after treatment with Sancao jiangya decoction of low,middle,high concentration by tailartery blood pressure measurement for conscious rats.Results: The blood pressure was decreased at 2 hours after drug taken,there were significant dose-effect relationship between the antihypertensive effect and the low,middle,high dose.At 4h after drug taken,the high,middle dose had dose-effect correlation,but the low-dose had no antihypertensive effect.Further research on the middledose shows that the blood pressure reduced at 1h after drug taken,and the stable antihypertensive effect was keeping during 1-4h,the blood pressure began to rise at 6h,and got back to the level before drug taken at 8h.Conclusion: To choose the Middle-dose(10.4g crude drug/kg body weight) and 2h after drug taken is appropriate for SCD's use.This result laid a substantial foundation for further research on effects evaluation and mechanism of antihypertensive medicine.

OBJECTIVETo construct a retrovirus-mediated expression system carrying human SHP-1 gene to transfer SHP-1 gene in human breast cancer MDA-MB-231 cells.

METHODSThe full-length SHP-1 gene fragment was amplified by RT-PCR from the total RNA extracted from human breast cancer cell line MCF-7 over-expressing SHP-1 protein. The gene fragment was inserted into the vector pLNCX2 to construct the recombinant retroviral plasmid, which was transfected into the packaging cell PT67 via Lipofectamine2000. A cell line stably producing the virus was selected with G418. MDA-MB-231 cells was infected with the virus, and the expression of SHP-1 gene in the positive cell clone was detected with Western blotting.

RESULTSA 1.8 kb cDNA fragment of SHP-1 gene was obtained from MCF-7 cells and successfully inserted into the pLNCX2. A stable cell clone PT67/SHP-1 and virus supernatant were obtained. Expression of SHP-1 protein was detected in the cells infected with the virus.

CONCLUSIONThe recombinant retroviral vector carrying SHP-1 gene has been successfully constructed and MDA- MB-231/SHP-1 cell line expressing SHP-1 has been obtained to allow further functional study of SHP-1 in breast cancer.

Breast Neoplasms ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Cloning, Molecular ; Genetic Vectors ; genetics ; Humans ; Protein Tyrosine Phosphatase, Non-Receptor Type 6 ; biosynthesis ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Retroviridae ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection

OBJECTIVE The aim of the study was to characterize the pharmacokinetics (PK) and pharmacodynamics (PD) profile of cisatracurium in 0-2 years and 2-5 years old children patients with cheilopalatognathus, to find if there are some connections between the different muscle relaxation action and different PK procedure . METHODS 14 children patients were divided into two groups, ≤2 years and 2-5 years group, venous samples were taken before injection of a 0.15 mg·kg- 1 dose of cisatracurium and then at 2, 5, 10, 30, 60, 90, and 120 min. Cisatracurium plasma concentrations were determined by ultra- performance liquid chromatography/electrospray ionization/triple quadrupole tandem mass spectrometer system (UPLC/MS/MS). The degree of neuromuscular block was measured by train of four (TOF) testing. An indirect PK-PD link model with a sigmoid Emax model was established using Win Nonlin software. The model were applied to PK and PD data analysis, respectively. RESULTS The TOF monitor parameters showed that cisatracurium works very quickly, the onset time were (2.64± 0.93) min and (2.59 ± 0.90) min for ≤2 years and 2- 5 years group respectively. Young children ≤2 years have longer muscle blocking duration time (62.5 ± 6.01 min vs 53.86 ± 12.18 min) and slower recovery index (32.14±7.10 min and 27.43±10.63 min) than those children in group of 2-5 years. More children ≤2 years have postoperative complication than that in 2-5 children. PK parameters showed that there were no statistical differences in blood concentration and pharmacokinetic parameters. While the concentration of cisatracurium in muscle site calculated by using PK/PD model were higher and longer for ≤2 year children than that of 2-5 year children. This means that cisatracurium could stay at high concentration for a longer time in younger children' muscle tissue. CONCLUSION As a result young children tend to have postoperative complications related to slower muscle recovery action and increased concentration in skeletal muscle. So more careful observation and monitor are needed for younger children, our study could be of use in clinical practice for the administration of cisatracurium to children patients.

OBJECTIVETo investigate the in vivo inhibition of extract of Fructus lycii (FL) on the expressions of cathepsin B (Cat B) and cystatin C (Cys C) in high-fat diet and hydroquinone (HQ) induced model mice with age-related macular degeneration (AMD), and to explore the in vitro effects of lutein and zeaxanthin on hydrogen peroxide (H2O2,) induced expressions of matrix metalloproteinase 2 (MMP-2) and tissue inhibitor of metalloproteinase 2 (TIMP-2) on ARPE-19 cells.

METHODSFifty female 8-month-old C57BL/6 mice were recruited in this research. Ten mice fed with regular diet was taken as the age control group. The rest 40 mice were fed with high fat diet for 6 months, followed by adding HQ (0. 8%) in the drinking water for 3 consecutive months. Then the modeled mice were randomly divided into the model control group (n =10), the high (at the daily dose of 3.75 g/kg), middle (at the daily dose of 2.50 g/kg), and low dose (at the daily dose of 1.25 g/kg) FL groups, 10 in each group. The extract of FL at each dose was respectively administered to mice by gastrogavage for 3 successive months. By the end of the experiment, the mice were killed and their eyeballs were removed. The protein expressions of Cat B and Cys C were observed by immunohistochemical assay. The mRNA and protein expressions of Cat B and Cys C were detected by real-time PCR and Western blot respectively. The drug concentrations of H2O2, lutein, and zeaxanthin were screened and detected using the activity of cell proliferation. The protein expressions of MMP-2 and TIMP-2 were detected using Western blot.

RESULTSCompared with the age control group, the mRNA and protein expressions of Cat B and Cys C were significantly higher in the in vivo model control group (P <0.05, P <0.01). The mRNA expressions of Cat B and Cys C were weaker in the middle and high dose FL groups than in the model control group (P <0. 05, P <0. 01). In in vitro cells, lutein and zeaxanthin could down-regulate the protein expressions of MMP-2 and TIMP-2 in H202 induced ARPE-19 cells (P <0. 05, P <0. 01).

CONCLUSIONSExtract of FL could down-regulate the high protein expressions of Cat B and Cys C in high-fat diet and HQ induced model mice. Lutein and zeaxanthin could down-regulate the protein expressions of MMP-2 and TIMP-2 in H202 induced ARPE-19 cells.

Animals ; Cathepsin B ; metabolism ; Cystatin C ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Female ; Hydrogen Peroxide ; Lutein ; pharmacology ; Macular Degeneration ; prevention & control ; Matrix Metalloproteinase 2 ; metabolism ; Mice ; Mice, Inbred C57BL ; Pigment Epithelium of Eye ; drug effects ; metabolism ; Tissue Inhibitor of Metalloproteinase-2 ; metabolism ; Xanthophylls ; pharmacology ; Zeaxanthins