Objective To detect the level of nitric oxide (NO) and amino acids in cerebrospinal fluid and serum of epilepsy children in different time. Methods The level of NO and amino acids in cerebrospinal fluid and serum of epilepsy children in different time were determined with nitrate reductase and high performance liquid chromatography (HPLC) with fluorimetric detection. Results The level of NO2 - increased 30 min, 60 min, 120 min after seizures (P<0.01), but dropped to normal 180 min after seizures (P>0.05). The level of glutamate (Glu) significantly increased in all epilepsy groups (P<0.05). The level of glycine (Gly) dropped gradually in all epilepsy groups (P<0.05).Conclusion NO may participate in the generation of seizures and the injury process to brain. The imbalance between excitatory and inhibitory amino acids may play an important role in the pathogenesis of epilepsy. The toxicity of excitatory amino acids may be the main injury to the brain.

Objective To investigate the dynamic changes of the spinal cord during neck flexion in Hirayama disease for diagnosis.Methods MRI examinations in neutral neck position and a fully flexed neck position were performed on 18 cases of Hirayama disease and 31 young normal control subjects.We measured an antero-posterior diameter(APD)and transverse diameter(TD)of the cervical cord at the superior margin of the C6 vertebral body for each position,and investigate the dynamic changes.The different in frequency of these findings between the control and patient groups was examined by means of the x~2 test.The group means were compared by independent-sample t-test.Significance was defined as P

OBJECTIVETo observe dynamically the development of fetal long bone and detect the expression and distribution of HIF-1alpha,to investigate the expression pattern and possible effects of hypoxia inducible factor-1alpha (HIF-1alpha) in fetal long bone development of mouse.

METHODSE12.5, E13.5, E14.5, E15.5, E16.5 and E17.5 pregnant C57BL6 mice were sacrificed. After sacrifice, the embryos were delivered by caesarean section. The development of fetal long bone was dynamically observed by stereoscopic microscope, and the distributional expression of HIF-1alpha protein was detected by using method of immunohistochemistry. The expression of HIF-1alpha mRNA and osteoblast marker gene at various stage were also detected by using methods of reverse transcription-polymerase chain reaction (RT-PCR).

RESULTSThe cartilaginous long bone began to form and joints outline arised at E13.5, then the primary ossification center was observed at E14.5, showing opaque ossification under stereoscopic microscope,and then the osteogenesis expanded and extended to both sides. Immunohistochemistry demonstrated lots of HIF-1alpha protein positive chondrcytes in the center of primary ossification at E14.5, then they decreased dramatically. HIF-1alpha mRNA expressed at high level from E13.5 to E15.5, and then decreased to low level.

CONCLUSIONFetal long bone development pattern appeared to be endochondral osteogenisis process, existing hypoxia microenviroment may increase HIF-1alpha mRNA expression and thus initiate the cascade of endochondral osteogenisis.

Animals ; Bone Development ; Female ; Hypoxia-Inducible Factor 1, alpha Subunit ; analysis ; genetics ; physiology ; Immunohistochemistry ; Male ; Mice ; RNA, Messenger ; analysis

OBJECTIVETo investigate the degradation of artificial basement membrane (matrigel) co-cultured with oral carcinoma-associated fibroblasts (CAFs) and its possible mechanism.

METHODSCAFs and normal fibroblasts (NFs) were incubated on matrigel for 24, 48, 72 h. Equivalent amounts of conditioned medium were collected and assayed for total protein, hydroxyproline and matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9) activity by gelatin zymography.

RESULTSOral CAFs were superior to oral NFs in total protein and hydroxyproline density, CAFs present more pro-MMP-2 and activated MMP-2.

CONCLUSIONCAFs were superior to NFs in degradation of matrigel. CAFs might play a key role in the reconstitution of extracellular matrix and the progression of tumor.

Basement Membrane ; Coculture Techniques ; Enzyme Precursors ; Fibroblasts ; Gelatinases ; Humans ; Matrix Metalloproteinase 2 ; Matrix Metalloproteinase 9 ; Membranes, Artificial ; Mouth Neoplasms

Objective To investigate drug resistance genes and epidemic characteristics of β-lactamase carried by carbapenem-resistant Acinetobacter baumannii (CRAB) in the respiratory intensive care unit(RICU) in a hospital.Methods Clinically isolated CRAB from RICU patients in October-December 2015 were collected.Five drug resistance genes (KPC-2,IMP,VIM,NDM-1,OXA-23) were specifically amplified by polymerase chain reaction (PCR),amplified products were performed agarose gel electrophoresis and sequencing analysis,the homology was analyzed with pulsed-field gel electrophoresis (PFGE).Results A total of 22 CRAB strains were isolated in October-December 2015,19 (86.36％) of which were isolated from sputum.The resistance rate of 22 CRAB strains to compound sulfamethoxazole was 59.09 ％,resistance rate to minocycline was 9.09 ％,all were sensitive to polymyxin B,resistance rates to other antimicrobial agents were more than 80％.Three kinds of resistance genes KPC-2,IMP and NDM-1 were not found by PCR amplification,positive rates of VIM and OXA-23 were both 100％.PFGE homology analysis revealed that 22 strains were divided into 13 different types,each type contained 1-5 strains,9 types(69.23％) contained only 1 strain respectively,the other 4 types (30.77％) contained 2-5 strains.A5,A7,and A8;A9,A11,A14,A19 and A22;A4,A10 and A12;A16 and A18 were of the same type respectively.Conclusion The main types of β-lactamase-resistant genes of CRAB in RICU are VIM and OXA-23.Homology analysis shows a small parts are of the same clone strains,which reveals epidemic of a small scale.

Objective To investigate epidemical situation of drinking-tea fluomsis in Qinghai province, in order to understand diet structure to provide the scientific basis for the prevention and control and the scientific research. Methods In 2007, according to "Scheme for Survey on Epidemical Drinking-tea Fluorosis", we carried out an customized investigation in 28 counties, 3 townships sampled in each county, 2 villages in each township, 50 adults and 50 school-age children in each village; at same time, 1 monk temple was sampled in each county, 50 clergy adults and 50 children in each temple. Then we investigated the resident income, the diet structure, the brick tea consumption and so on, and applied Dean method to diagnose dental fluorosis. The patient number estimated based on the survey result. Results ①Yeady per capita income of people was mostly 500 - < 1000yuan, next 1000 - < 3000 yuan; economic income in pasturing area was higher than that of agriculture, half area and half agriculture and half pastoral region and township. ②Staple food was bread flour primarily in the animal husbandry agricultural half pastoral area,next were the roasted barley and the rice;the bread flour was the principle food in the agricultural region and the cities,next were the rice and the roasted barley;among non-staple food,meat came fimt and milk foHowed,egg the last.③The frequently edible vegetables Was potato.cabbage and greenpepper,eaten by a majority of people[62.82%(6497/10 343)];as for fruits,apple,pear and orange was primarily consumed,75.95%(7856/10 343)of people ate less than 10 times every month.④Sixty-one thousand nine hundred and ninety-nine residents were registered,153 335 kg ofbrick tea was consumed in villages and towns,2.47 kg per person;in 1001 monks investigated,4120 kg of brick tea was consumed every year,4.12 kg per person.⑤Detection rate of adult dental fluorosis in the villages and towns was 24.11%(2494/10 343),that of the children was 24.38% (3012/12 355);detection rate of dental fluorosis in monks was 26.13%(203/777),that of the children was 39.73% (89/224).⑥Detection rate of adult skeletal fluorosis in villages and towns was 15.60%(17/109);that of monks was 4.88%(2/41).⑦The 95%confidence limit estimated a total number of dental fluorosis patients were 1 084 306- 1 134 170 persons.the median Was 1 109 238 persons;the 95% confidence limit estimated a total number of skeletal fluorosis patients were 309 177-758 199 persons,the median was 533 688 persons.Conclusions Qinghai province has a great quantity of brick tea consumption,having lots of people with drinking-tea fluorosis which is in severe degree.The resident food structure was monotonous and mostly transpolted from other region.

OBJECTIVETo study the optimum technology of extracting bioactive component named geniposidic acid in Eucommia Ulmiodes by orthogonal test.

METHODAn orthogonal test table L16(4(5)) was used with the factors of extracting time, temperature, times, ethanol consistency and the ratio of solid to liquid. The content of geniposideic acid was determined by RP-HPLC.

RESULTThe optimum technology was: temperature 50 degrees C, time 1 h, once, consistency 80%, and the ratio of solid and liquid 1:12. Under the condition, the best material of Eucommia Ulmiodes for extracting geniposidic acid was chosen.

CONCLUSIONThe technology can provide some theoretic advice for industry.

Chromatography, High Pressure Liquid ; Ethanol ; Eucommiaceae ; chemistry ; Glucosides ; analysis ; isolation & purification ; Iridoid Glucosides ; Iridoids ; analysis ; isolation & purification ; Plant Bark ; chemistry ; Plants, Medicinal ; chemistry ; Technology, Pharmaceutical ; methods ; Temperature ; Time

Classical Swine Fever Virus (CSFV) E2 protein eukaryotic expression plasmid pVAXE2 was constructed. The plasmid pVAXE2 was transformed into Salmonella choleraesuis C500 (S. C500) attenuated vaccine strain by electroporation to generate Salmonella choleraesuis engineering strain S. C500/pVAXE2. The characterization of S. C500/pVAXE2 in morphology, growth, biochemistry and serology indicated that it retained the same properties as its original strain S. C500 with exception of kanamycin resistance originated from the plasmid pVAXE2. The plasmid stable in the bacteria after 15 passages. Kunming mice and rabbits were vaccinated three times at two weeks interval with S. C500/pVAXE2 in oral and intramuscular routes at the dosage of 1 x 10(8) CFU for mice and 2 x 10(9) CFU for rabbits each time. The specific antibody response against CSFV and Salmonella choleraesuis was detected by ELISA. Two weeks after the third boost the immunized rabbits were challenged with 20 ID50 of hog cholera lapinized virus (HCLV), followed by a virulent strain of Salmonella choleraesuis two week later than HCLV challenge. The results showed that all immunized mice and rabbits produced significant antibodies against CSFV and Salmonella choleraesuis, and the immunized rabbits demonstrated the effective protection against the challenge of HCLV and virulent Salmonella choleraesuis. These results indicated the potential of developing multiplex swine DNA vaccine by using this bacteria as the vector.
Animals ; Classical Swine Fever ; immunology ; prevention & control ; virology ; Classical swine fever virus ; genetics ; immunology ; Mice ; Rabbits ; Salmonella arizonae ; genetics ; Swine ; Vaccines, DNA ; immunology ; Viral Envelope Proteins ; biosynthesis ; genetics ; immunology ; Viral Vaccines ; immunology

Objective To stimulate the expansion of natural killer cells (NK) in vivo, to achieve high expression of Flt-3 ligand ( FL) in the mouse liver by hydrodynamic injection technology , to obtain high purity of NK for cell therapy and then to purify the spleen lymphocytes by MicroBeads sorting ( MACS ) technique.Methods C57BL/6 mice were repeatedly treated with hydrodynamic injection of FL expression plasmid .Then, surface markers of splenic NK were analyzed by flow cytometry and purified by MACS technique .Results Compared with mice treated with one hydrodynamic injection, the spleen of mice treated with two hydrodynamic injections showed significant variation, with the absolute num-ber of lymphocytes increased (6.02 ±1.15) times, the proportion of NK subsets increased (2.07 ±0.39) times, and the absolute number of NK subsets increased (12.49 ±2.39) times.Cell surface marker detection confirmed that NK activity and surface markers did not change significantly .NK with a purity of about (83.81 ±0.92)％ was obtained by the combination of two magnetic beads sorted with CD 5 ( Ly-1) MicroBeads Positive selection and EasySep TM Mouse NK Cell Isolation Kit negative selection .Conclusion NK can be expanded in the spleen of mice by hydrodynamic injection of FL plasmid without influence on the NK activity and surface markers.The combined use of CD5 (Ly-1) MicroBeads positive selection with EasySep TM Mouse NK Cell Isolation Kit negative selection can effectively remove T , B and other miscellaneous cells, thereby contributing to obtaining high-purity NK. This study provides good reference for the immunotherapy of tumor cells.

Objective :To explore value of high sensitive cardiac troponin I (hs-cTnI) in monitoring cardiac toxicity of patients with leukemia undergoing anthracycline chemotherapy .Methods :A total of 160 patients with acute myeloid leukemia ,who were treated in our department from Jan 2013 to Dec 2016 ,were enrolled .Patients were randomly divided into routine chemotherapy group and Dexrazoxane group (received Dexrazoxane on 30 min before chemo-therapy) ,both groups were treated for six periods (21d×6).Serum hs-cTnI level and incidence of cardiac toxicity events before and at different time points after chemotherapy were observed and compared between two groups .In-cidence rate of cardiac toxicity events was compared between groups with different serum hs-cTnI level.ROC curve was used to analyze predictive value of serum hs-cTnI for cardiac toxicity .Results :Compared with before chemo-therapy ,after chemotherapy ,there was significant rise in serum hs-cTnI level in each period in two groups ( P＝0.001 all) ,and it rose along with time passed .Compared with routine chemotherapy group from second period , there was significant reduction in serum hs-cTnI level [2nd21d :(7.59 ± 0.85) 10－2ng/ml vs.(7.27 ± 0.86) 10－2ng/ml ,] in Dexrazoxane group , P＜ 0.05 or ＜ 0.01. Total incidence rate of cardiac toxicity events of Dexrazoxane group was significantly lower than that of routine chemotherapy group (5.00% vs.25.00%) ,P＝0.001. Incidence rate of cardiac toxicity event in hs-cTnI＞0.100ng/ml group was significantly higher than that of hs-cTnI≤0.100ng/ml group (31.25% vs.4.17%) ,P＝0.001. ROC curve suggested that AUC was 0.825 ,cutoff point was 0.121 ng/ml ,sensitivity and specificity was 90.27% and 92. 35% respectively for serum hs-cTnI predicting cardiac toxicity in these patients .Conclusion : Serum hs-cTnI level rises in early period of chemotherapy in patients with leukemia un- dergoing anthracycline chemotherapy , so it possesses high predictive value for cardiac toxicity events .