A method was developed for determining residual dexamethasone and betamethasone in swine liver by HPLC-MS/MS with isotope dilution. The samples were digested by β-glucuronidase/aryl sulfatase, and extracted) with acetonitrile. Further cleanup was performed on C_(18)) cartridge and liquid-liquid extraction with sodium) carbonate solution. Then the supernatant was dried under nitrogen and residues were dissolved in mobile) phase. The samples were analyzed by HPLC-MS/MS on a Hypercarb C_(18)) column with a mixture of acetonitrile-water-formic acid as the mobile phase. The samples were quantified with the internal standard calibration curve method using isotope dilution. The limit of detection for dexamethasone and betamethasone in swine liver was 0.12 μg/kg and 0.14 μg/kg, respectively. The limits of quantification were 0.42 μg/kg and0.47 μg/kg), respectively. The average recoveries were 97.3%-111.0%, the intra-assay relative standard deviations were 1.85%-5.65% and the inter-assay relative standard deviations were 2.8%-8.0% at spiked levels of 0.75-2.00 μg/kg. There was a good linear correlation (the correlation coefficient is above 0.9997) between the ratio of peak areas of quantitative ion-pair to internal standard and concentration of analyte) in the range of 10-500 μg/L.

The expert system is an important field of the artificial intelligence. The traditional interface of the expert system is the command, menu and window at present. It limits the application of the expert system and embarrasses the enthusiasm of using expert system. Combining with the study on the expert system of network fault diagnosis, the natural language interface of the expert system has been discussed in this article. This interface can understand and generate Chinese sentences. Using this interface, the user and field experts can use the expert system to diagnose the fault of network conveniently. In the article, first, the extended production rule has been proposed. Then the methods of Chinese sentence generation from conceptual graphs and the model of expert system are introduced in detail. Using this model, the network fault diagnosis expert system and its natural language interface have been developed with Prolog.

Objective To investigate the effects of baicalin on morphine-induced behavioral sensitization.Methods Locomotor activity was measured for 2h after administration with baicalin in mice.Hyperlocomotion induced by acute morphine (10 mg· kg-1,ip) and behavioral sensitization induced by repeated morphine were established.The level of dopamine of ventral tegmental area(VTA) and prefrontal cortex(PFC) in mice was tested by ELISA assay.Results Baiealin inhibited significantly both locomotor activity in mice (control (1095.8 ± 174.5) times,baicalin (899.6± 187.2),(724.2± 221.4),(609.1 ± 154.6) times ; P＜ 0.01) and hyperlocomotion induced by acute morphine(model (1518.2± 185.8) times,baicalin (1385.4±224.2),(1205.1 ± 174.6),(1100.3±235.1) times ; P＜0.01).Similar inhibition was also seen in the development and expression of morphine-induced behavioral sensitization(model(2096.2±304.6) times,baicalin (2004.2 ± 218.5),(1998.7-± 224.3),(1836.1 ± 233.5) times,P＜ 0.05 ; model (2124.2 ± 189.6) times,baicalin (1922.2± 314.7),(1524.1±289.2),(1477.4± 219.3) times,P＜0.01).Baicalin inhibited dopamine release in VTA and PFC of morphine-sensitized mice(model(457.6± 92.1,589.2 ±102.5) μg · L-1,baicalin(391.1±56.8) μg · L-1,(448.6± 99.3) μg · L-1; (324.5±66.2) μg · L-1,(368.7±45.9) μg · L 1 ; (234.3± 52.6) μg · L-1,(305.3±84.1) μg · L-1 ; P＜0.01,P＜0.01).Conclusion Baicalin inhibits the development and the expression of morphine-induced behavioral sensitization in mice,and this effect is related to the inhibition of dopamine release in VTA and PFC of mice.

BACKGROUND:Traditional bone cement in the treatment of osteoporotic vertebral compression fracture easily induces heat dissipation effect, leakage, big difference in mechanical strength with the surrounding tissue, which greatly affects treatment effect of osteoporotic vertebral compression fracture. This prospective self-controled open-label clinical trial is designed to analyze the effectiveness of a novel high viscosity bone cement for osteoporotic vertebral compression fractures.
METHODS/DESIGN:This prospective self-controled open-label clinical trial wil be performed in the Yixing Hospital Affiliated to Jiangsu University of China. High viscosity bone cement wil be implanted in patients with osteoporotic vertebral compression fractures by percutaneous vertebroplasty. Immediate outcomes: Pain symptom of patients before and after implantation of high viscosity bone cement, and Visual Analogue Scale score. Middle- and long-term outcomes: The recovery of spinal function, Oswestry dysfunction index questionnaire, vertebral body height, bone cement leakage rate, Barthel index and SF-36 quality of life scale score.
DISCUSSION: This trial wil provide a clinical basis for the treatment of osteoporotic vertebral compression fractures with high viscosity bone cement.
ETHICS APPROVAL: This trial has been approved by the Medical Ethics Committee, Yixing Hospital Affiliated to Jiangsu University (Approval number 0136). Patients and their family members have signed the informed consent.

Objective To explore the indications for percutaneous renal biopsy of asymptomatic hematuria in children. Methods The renal pathological types of 485 children with asymptomatic hematuria were analyzed retrospectively. According to the degree of hematuria and whether or not combined with proteinuria, the children were divided into microscopic hematuria group, gross hematuria group and hematuria with proteinuria group. The microscopic hematuria group was further divided into urine red blood cell<15/HPF group, (15～30)/HPF group, and >30/HPF group according to hematuria degree. Results In 227 males and 258 females with the average age of 7.23±2.93 years, there were 318 cases in microscopic hematuria group, in which the most common pathological types were minor lesions (64.8%), followed by focal glomerular lesions (16.7%) and focal segmental glomerulosclerosis (8.2%). There were 119 cases in gross hematuria group, in which the most common pathological types were also minor lesions (26.1%), followed by IgA nephropathy (24.4%) and mesangial proliferative glomerulopathy (20.2%). There were 48 cases in hematuria with proteinuria group, in which the most common pathological types were IgA nephropathy (29.2%) and minor lesions (29.2%). The distribution of the pathological types among microscopic hematuria group, gross hematuria group and hematuria with proteinuria group were statistically different (χ2=152.03, P<0.001). In three groups, microscopic hematuria group had the highest proportion of minor lesions, while gross hematuria group and hematuria with proteinuria group had higher proportion of IgA nephropathy and mesangial proliferative glomerulonephritis . In microscopic hematuria group, there were 149 children with urine red blood cell<15/HPF, 96 with urine red blood cell (15～30)/HPF, and 73 with urine red blood cell >30/HPF. There was no difference in pathological types among three sub-groups (χ2=15.18, P=0.51), and mild lesions were the most common pathological types in each group. Conclusion Renal biopsy should be performed at earliest possible time to make pathological diagnosis in asymptomatic hematuria children with gross hematuria or proteinuria.

OBJECTIVE:To study induction effect of euphornin on the apoptosis of cervical cancer Hela cells and its mechanism. METHODS:The cervical cancer Hela cells were divided into blank control group,cisplatin group(positive control, 10 mg/L) and euphornin low-dose,medium-dose and high-dose groups (50,100,200 mg/L). They were treated with relevant medicine. The inhibitory effect of Hela cells proliferation was tested by MTT assay after 24,48,72 h of medicine treatment. The apoptotic rate of Hela cells was measured by flow cytometry after 48 h of medicine treatment. Morphology of nucleus was detected by Hoechst 33258 staining. The protein expression of Cyt-C,Bcl-2,Bax,Caspase-3,Caspase-8,Caspase-9 and Caspase-10 were detected by Western blot assay. RESULTS:Compared with blank control group,inhibitory rate of cell proliferation and cell apoptosis rate were increased significantly in cisplatin group and euphornin groups(P＜0.05 or P＜0.01),and obvious staining, deformation,shrinking,fragmentation or apoptotic bodies was found in nucleus. Compared with blank control group,the protein expression levels of Cyt-C,Caspase-8 and Caspase-9 in euphornin low-dose,medium-dose and high-dose groups were increased significantly,while the protein expression level of Bcl-2 and Bcl-2/Bax ratio were decreased significantly(P＜0.05 or P＜0.01);the protein expression levels of Bax,Caspase-3 and Caspase-10 in euphornin medium-dose and high-dose groups were increased significantly(P＜0.05 or P＜0.01). CONCLUSIONS:Euphornin can significantly inhibit the proliferation of Hela cell and promote cell apoptosis,the effect of which will be achieved by activating the Caspase-dependent mitochondrion apoptosis pathway.

Objective To evaluate the effects of ganglioside GM1 on hypoxic ischemic brain damage (HIBD) in neonatal rats and on the expression of potassium-chloride cotransporter 2 (KCC2) in hippocampus.Methods Seven-day-old Sprague-Dawley (SD) rats (n=72) were randomly divided into a sham group,an HIBD group and a ganglioside GM1 group.Each group was further divided into a 3 d subgroup and a 21 d subgroup according to the different detection index (n=12).Rat HIBD models were prepared according the Rice-Vannucci method.After HIBD,the ganglioside GM1 group was given ganglioside GM1 20 mg/ (kg ·d) by intraperitoneal injection for 3 d continuously.2-,3-,5-triphenyltetrazolium chloride (TTC) was preformed to evaluate the area of cerebral infarction of HIBD in each 3 d subgroup.Spontaneous activity recorder was used to observe the locomotor activity of the rats in the 21 d subgroups.Morris water maze test was conducted for assessment of rats' learning and memory abilities in the 21 d subgroups.Western blot analysis was employed to determine the alterations in KCC2 expression in hippocampus in all the 3 d and 21 d subgroups.Results Compared with the HIBD group (28.6％±5.2％),the ratio of cerebral infarction volume in the ganglioside GM1 group (11.3％±2.4％) was significantly reduced (P＜0.05).Compared with the HIBD group (289.6±61.3),the number of locomotor activities within 2 h in the ganglioside GM1 group (412.1±66.8) was significantly increased (P＜0.05).Compared with the HIBD group,the escape latency was significantly reduced,but the percentage time of target quadrant and the number of crossing the platform were significantly increased in the ganglioside GM1 group (P＜0.05).Three days and 21 days after HIBD,the expression of KCC2 in the ganglioside GM1 group was significantly higher than that in the HIBD group (P＜0.05).Conclusion Ganglioside GM1 may have a significant protective effect on HIBD in neonatal rats,and its mechanism may be related to regulation of the expression of KCC2 in hippocampus.

OBJECTIVETo compare the results of femoral head replacement (FHR) and total hip replacement (THR) in treatment of subcapital femoral neck fractures (SFNF).

METHODSBetween May 1987 and July 1998, 56 elderly patients (6 5-90 years; average 73.5 years) with SFNF were treated with prosthetic replacement. Six cases were treated with unipolar FHR, 18 cases with Bateman bipolar FHR, and 32 cases with Bateman bipolar THR. All domestic prostheses were installed with cement.

RESULTSThere was no significant difference between the 2 groups in operating time and blood transfusion. Forty-nine patients were followed-up for an average of 5 years and 10 months. No wound infection or death was related to surgery. Complications in Group FHR were significantly higher than that i n Group THR.

CONCLUSIONSSince FHR is difficult to fit the bony acetabulum, it is only indicated for senile cases with poor conditions. However, the bi polar THR installed with cement is indicated for most elderly patients. Since th e femoral head and acetabulum can fit each other completely, it is more stable for taking weight-bearing earlier with less complications.

Aged ; Aged, 80 and over ; Arthroplasty, Replacement, Hip ; Bone Cements ; Female ; Femoral Neck Fractures ; diagnostic imaging ; surgery ; Hip Prosthesis ; Humans ; Male ; Radiography ; Treatment Outcome

To prepare a soluble human extracellular III domain of Flt1 and analyze its biological activity. The gene encoding extracellular domain III of Flt-1 was cloned into the expression vector pAZY by RT-PCR from human umbilical vein endothelial cell (HUVEC), and induced to express in Escherichia coli by low phosphoric medium, the product was purified by E-tag affinity chromatography. SDS-PAGE and Western blotting analysis showed that Flt-1 gene domain III gene was expressed in E. coli and the yield of the soluble fusion protein was about 1.10 mg/L. Enzyme-Linked ImmunoSorbent Assay (ELISA) revealed that the Flt-1 domain III was able to bind to VEGF165 dose-dependently. Monolayer denudation assay and Transwell assay showed that the fusion protein could inhibit HUVECs migration induced by conditional medium with 50 ng/mL VEGF165 and 100 ng/mL bFGF. In conclusion, Flt-1 gene domain III gene has been successfully cloned and expressed in E. coli, which will be useful in both the research on the function of Flt-1 gene domain III and preparation of anti-Flt-1 monoclonal antibody in the future.
Cloning, Molecular ; Endothelial Cells ; cytology ; metabolism ; Escherichia coli ; genetics ; metabolism ; Extracellular Space ; metabolism ; Genetic Vectors ; genetics ; Humans ; Recombinant Fusion Proteins ; genetics ; isolation & purification ; metabolism ; pharmacology ; Solubility ; Umbilical Veins ; cytology ; Vascular Endothelial Growth Factor Receptor-1 ; genetics ; isolation & purification ; metabolism

Intensive cancer treatment with drug combination is widely exploited in the clinic but suffers from inconsistent pharmacokinetics among different therapeutic agents.To overcome it,the emerging nanomedicine offers an unparalleled opportunity for encapsulating multiple drugs in a nano-carrier.Herein,a two-step super-assembled strategy was performed to unify the pharmacokinetics of a pep-tide and a small molecular compound.In this proof-of-concept study,the bioinformatics analysis firstly revealed the potential synergies towards hepatoma therapy for the associative inhibition of exportin 1(XPO1)and ataxia telangiectasia mutated-Rad3-related(ATR),and then a super-assembled nano-pill(gold nano drug carrier loaded AZD6738 and 97-110 amino acids of apoptin(AP)(AA@G))was con-structed through camouflaging AZD6738(ATR small-molecule inhibitor)-binding human serum albumin onto the AP-Au supramolecular nanoparticle.As expected,both in vitro and in vivo experiment results verified that the AA@G possessed extraordinary biocompatibility and enhanced therapeutic effect through inducing cell cycle arrest,promoting DNA damage and inhibiting DNA repair of hepatoma cell.This work not only provides a co-delivery strategy for intensive liver cancer treatment with the clinical translational potential,but develops a common approach to unify the pharmacokinetics of peptide and small-molecular compounds,thereby extending the scope of drugs for developing the advanced com-bination therapy.