Objective To establish a HPLC fingerprint analysis method of salt-prepared Cortex Phellodendri. Methods C18 column was used, with gradient elution by the mobile phase consisted of acetonitrile-0. 1% phosphoric acid (contained 0. 2 % triethylamine). The detection wavelength was at 230 nm, and the flow rate was at 0. 8 mL/min. Results Seventeen char-acteristic peaks were selected and the fingerprints on HPLC was set up. Conclusion HPLC fingerprint method is repro-ducible, accurate and stable, and can be used for the quality control of salt-prepared Cortex PheUodendri.

Objective To study the immunohistochemistry change of neuronal nitric oxide synthase(nNOS) during the chronic spinal cord compression. Methods 18 healthy rabbits were randomly divided into normal,control and compressed groups.Membranous sac filled with cardiografin was applied to produce an animal model of chronic spinal cord compression.The sac was gradually enlarged resulting in chronic spinal cord compression in compressed groups for 12 weeks.Nissl's staining was applied to observe histopathological change and immunohistochemistry to nNOS change. Results The damage of motorneurons in the compressed block of compressed group was observed.No histopathological change was observed in normal and control groups.The number of nNOS positive motor neurons in the compressed block of compressed group was higher than that in the blocks of other groups.Conclusion\ The NO synthesis increased in chronic spinal cord injury.\;[

Objective To establish a HPLC fingerprint analysis method of salt-prepared Cortex Phellodendri. Methods C18 column was used, with gradient elution by the mobile phase consisted of acetonitrile-0.1 % phosphoric acid (contained 0.2 % triethylamine). The detection wavelength was at 230 nm, and the flow rate was at 0.8 mL/min. Results Seventeen characteristic peaks were selected and the fingerprints on HPLC was set up. Conclusion HPLC fingerprint method is reproducible, accurate and stable, and can be used for the quality control of salt-prepared Cortex Phellodendri.

Objective To compare treatment and efficacy of thoracolumbar fractures by using three different screw fixations:traditional approach,the vertebral side clearance into the road and percutaneous pedicle. Methods A total of 82 single segmental thoracic lumbar fractures cases hospitalized from March 2011 to March 2014 ,with male 67 cases ,female 15 cases ,and average age(33.7+/-12.5)years old. Patients were randomly divided into three groups:traditional approach group (n = 23),operation through paraspinal muscle gap group (n = 30),percutaneous group(n = 29). These following indicators will be compared in three groups:duration of operation ,intraoperative blood loss ,intraoperative fluoroscopy time ,postoperative flow ,VAS scores before and after operation and Oswestry disability index , difference of spinal sagittal position Cobb′s Angle. Results Compared with the traditional approach group ,operation through paraspinal muscle gap group and percu-taneous group have obvious advantages in duration of operation,intraoperative blood loss,postoperative flow,VAS scores before and after operation ,the Oswestry disability index. Additionally ,above mentioned three surgical methods could recover kyphosis deformity ,and there was no statistically significant difference among three groups (P > 0.05). Conclusion In the treatment of monosegmental thoracolumbar fractures ,compared with traditional approach ,operation through paraspinal muscle gap and percutaneous pedicle screw internal fixation have more advantages which includes fewer trauma,less bleeding,faster recovery and lower incidence of postoperative low back pain.

In 34 hypothalamic slices of rats, spontaneous discharging of 63 paraventricular neurons was extracellulariy recorded by glass microelectrode. When the slices were perfused with artificial cerebrospinal fluid (ACSF) containing noradrenaline (NA) (10-6 mol/L), firing rates of 20 units significantly increased, those of 8 units decreased or even ceased, and those of 35 units, not affected. When low Ca2+ high Mg2+-ACSF was applied to block synaptic transmission, of 20 units which were excited by NA, 14 units still had excitatory responses and 6 units had no significant response to NA. When synaptic transmission was blocked, of 8 units which were inhibited by NA, 7 units still had inhibitory responses and only one unit had no significant response to NA. The results of this experiment strongly suggest the existence of direct effects of NA on paraventricular neurons.

Objective To evaluate the expression of KAII (CD82) gene inhibited by small interfering RNA (siRNA) in human pancreatic cancer cell line T3. Methods Four sequences of siRNA including A, B,C, D were designed, which were based on the KAI1 gene sequence using online RNA interfering designing software and lentivirus vector was built. Then they were used to transfect T3 cells by liposome 2000 and virus titer was determined. Empty vector containing siRNAd1 lentivrus particle ( MOI =5) was also used to infect T3 cells. The expression of CD82 mRNA was detected by real-time PCR. Results The expression of CD82 mRNA in normal control group, empty vector group, A group, B group, C group, D group were 1. 398 ±0.242,1. 311±0.048, 0. 664 + 0. 093, 0. 345 ± 0. 032, 0. 641 ± 0. 049 and 0. 147 ± 0. 049, respectively, the difference between the expression of CD82 mRNA in empty vector group and that of A, B, C, D groups was significant (P＜0.01 ). Conclusions RNAi was able to inhibit the expression of KAI1 gene CD82 in human pancreatic cancer cell line T3.

Objective To investigate the effect of alpha 1-antitrypsin (A1AT) concerning in reducing the injury of transplanted islets by pancreas exocrine cells and promoting proliferation of the pancreas B cells.Method The pancreases of mice were digested with collagenase,islets were isolated artificially,and pancreatic exocrine cells were collected.In purified islet group (n =6),100 islets were seeded into a 6 well culture plate.In experimental group(n =6),100 islets were co-cultured with equal volume of pancreas exocrine cells,and 0.5 mg/mL A1AT was added into a 6-well culture plate.In control group(n =6),100 islets were co-cultured with equal volume of pancreas exocrine cells.After 48 h,insulin content of islets in each well and trypsin concentration in the supernatant of each well were measured.The islets were cultured in low sugar and high sugar 1640 medium,then glucose stimulated insulin secretion (GSIS) test was carried out.In vivo,8-9-week old male BALB/C mice were induced with STZ (190 mg/kg body weight,i.p) to establish the diabetic model and randomly divided into two groups.In experimental(n =10) and control(n =10) groups,250 islets and the equal volume of pancreatic exocrine cells were transplanted into different regions of left kidney subcapsule,resepctively.The experimental group was injected with A1AT (83 mg/kg,qd,i.p) for 28 days after operation,and the control group was injected with the same amount of normal saline (qd,i.p) for 28 days.Both two groups were given EDU (5 μg/g,qd,i.p) for 28 days.The blood glucose level was monitored continually.Nephrectomies were performed after 28 days.The expression of anti-amylase antibodies in the renal subcapsule was detected by immunohistochemical staining,and the proliferation of islet beta cells was examined using immunofluorescence staining.Result Insulin levels and insulin stimulation index in the control group were decreased as compared with those in the purified islet group; those in the experimental group were higher than in the control group,but lower than in the purified islet group.Trypsin concentration in the control group was increased as compared with the purified islet group,that in the experimental group was lower than the control group,but higher than in the purified islet group (all P＜0.01).After islets transplantation,the blood glucose levels in control and experimental groups were normal,but those in the control group recovered later than in the experimental group (P＜0.01).At 3rd day after nephrectomy,the blood glucose levels were ＞21 mmol/L in both two groups.A large number of anti-amylase antibody-positive cells were found in the renal subcapsule in the control group while little seen in the experimental group after 28 days.The immunofluorescence showed that the insulin +/EDU + B cells in the experimental group were more than those in the control group.Conclusion Conclusion Co-culture of islets and pancreatic exocrine cells with A1AT can prevent islet cells from damage caused by trypsin.A1AT could inhibit the secretion of pancreatic amylase from pancreatic acinar cells and promote proliferation of islet beta cells.

OBJECTIVE To investigate the resistance genes in HZB9055 strain of Enterobacter cloacae isolated from the PLA 98th Hospital,Huzhou District,Zhejiang Province,China.METHODS HZB9055 Strain of E.cloacae was isolated from the inpatient in May,2004,41 kinds(or groups) of genes were analyzed by PCR and verified by DNA sequencing.RESULTS In HZB9055 strain of E.cloacae,3 kinds of genes were positive including blaLAP,blaMIR and qnrS.The 38 kinds of rest genes were all tested negative.The blaLAP gene sequence including 858 nucleotides,which had the amino acid mutation in position 193 compared with LAP-1 type narrow-spectrum ?-lactamase(GenBank accession number:EF026092),and was nominated LAP-2(GenBank accession number:EU159120).CONCLUSIONS At least 3 kinds of resistance genes exist in HZB9055 strain of E.cloacae,and blaLAP-2 is a novel subtype of ?-lactamases gene.

Objective To establish a method of gold nanoprobe-based solution hybridization (GNBSH) to detect nucleic acid sequence-based amplification (NASBA) products for the rapid diagnosis of invasive aspergillosis (IA).Methods The Aspergillus specific 18S rRNA was amplified by NASBA and then the amplified products were hybridized with the gold nanoprobes which were modified with thiol compounds at the 5'end.Serum samples from 106 patients,including 14 with a definite IA,32 with suspected IA and 60 without IA,were detected by the established method,and the obtained results were compared with that of galactomannan (GM) test to evaluate its accuracy.Results The gold nanoprobes only hybridized with Aspergillus NASBA products but not other non-Aspergillus strains.The sensitivity,specificity and the area under the ROC curve (AUCROC) of the established GNBSH method for detecting 106 clinical samples were 82.61％ (38/46),81.67％ (49/60) and 0.890,respectively.The sensitivity,specificity and AUCROC of GM test were 56.52％ (26/46),83.33％ (50/60) and 0.723,respectively.Conclusion The established GNBSH method to detect Aspergillus NASBA products has high sensitivity and specificity and simple operation,which may be used to detect the infection of Aspergillus by clinical laboratories.

OBJECTIVE To investigate the produce of extended-spectrum ?-lactamases(ESBLs) and the presence of genotype of the ?-lactamases-encoding genes in Klebsiella pneumoniae isolated from the 98th Hospital of PLA,Huzhou,Zhejiang Province,China.METHODS Twenty-five strains of K.pneumoniae were isolated from the inpatients between Sep 2005 and Apr 2006.ESBLs were tested by phenotypic confirmatory tests recommended by CLSI.Twenty-one kinds of ?-lactamases genes of blaTEM,blaSHV,blaLEN,blaOKP,blaCTX-M-1 group,blaCTX-M-2 group,blaCTX-M-9 group,blaOXA-1 group,blaOXA-2 group,blaOXA-10 group,blCARB,blaPER,blaVEB,blaGES,blaLAP,blaDHA,blaACT/MIR,blaCMY/MOX,blaFOX,blaCMY/LAT,and blaACC were analyzed by PCR and verified by DNA sequencing.RESULTS In 25 strains of K.pneumoniae,the positive,negative,and "uncertainty" rates of ESBLs were 56.0%,20.0%,and 24.0%,respectively.The positive rate of genes of blaTEM,blaSHV,blaCTX-M-1 group,blaOXA-10 group,blaLAP,and blaDHA were 80.0%,4.0%,4.0%,80.0%,4.0% and 32.0%,respectively.The 15 kinds of rest genes were all tested negative.The total positive rate of 21 kinds of ?-lactamases gene was 92.0%.Among them,the blaLAP-2 gene sequence of the HZ12593 strain has been registered in GenBank(GenBank Accession Number: EU529981).CONCLUSIONS There are higher rate of ESBLs-producing strains in K.pneumoniae isolated from the inpatients,and at least 6 kinds of ?-lactamases gene existed.Both genes of blaTEM and blaOXA-10 group are the most common genotypes.Carring blaDHA Gene may influence the result of phenotypic confirmatory test for ESBLs in K.pneumoniae.