OBJECTIVE:To determine ampicillin concentration in human plasma by HPLC.METHODS:The chromatographic column was Waters Sunfire C18 with mobile phase consisted of acetonitrile and 0.05 mol?L-1 potassium dihydrogen phosphate(6.5∶93.5,V∶V)at a flow rate of 1.2 mL?min-1.The UV detection wavelength was 210 nm;the column temperature was 35 ℃ and the injection volume was 20 ?L.RESULTS:The linear range of ampicillin was 0.2～16.0 ?g?mL-1(r=0.999 9).The limit of quantization was 0.2 ?g?mL-1.The average methodological recovery was 99.88%～104.45%,and the average extraction recovery was 95.69%～100.56%.Both the inter-day RSD and the intra-day RSD were all less than 10%.CONCLUSION:This method is stable,rapid and sensitive,and suitable for the pharmacokinetic study for ampicillin preparation.

Flocculation of fermented broth with 1,3 -propanediol using component B of natural clarifier-Ⅱ was studied. Firstly, single factor tests and orthogonal tests were carried out. The results showed the main factors which influenced the flocculation and the optimal conditions were: pH6.0, flocculant B 0.01g/L , NaCl 0g/L, and the FR was 95.97% . The following results by filtration experiments, compared with the sample without pretreatment, indicated that the filtration speed of the flocculated sample was improved dramatically, and the weights of net and dry filter cakes were increased by 41.13% and 51.88% , respectively. Electrodialysis experiments showed that flocculation could accelerate desalting process, and could take the place of centrifugal pretreatment when it combined with the filtration.

Objective To retrospectively analyze the surgical methods and efficacy in 70 cases of type A aortic dissection patients over 65 years old.Methods From January 2005 to May 2012,70 type A aortic dissection patients over 65 years old received surgical treatment.Among them,there were 47 males aged 65 to 78 years old with mean 71,23 females,aged 65 to 72 years old with mean 68.55 cases were acute onset,while 15 cases were chronically onset.Different surgical methods were selected depend on patients' situations.We followed up all patients after discharged from hospital to continue to observe their health situation and evaluate the therapeutic effects.Results After surgery,eight patients died in the hospital,62 patients were recovered and discharged from the hospital.The mortality rate is 11.4％.During the follow up period from 3 to 72 months,there were no dead,aneurysm rupture and others severe complications.9 cases received endovascular graft exclusion within 6 months after discharged from hospital.The survival patients were satisfactory healed with their daily living activity resumed.Conclusion For over 65 years old patients with aortic dissection,the accurate and rapid selection of surgical method could improve the survival rate and the quality of life with a lower occurrence rate of complications.

Objective To investigate the factors influencing the prevalence of echinococciasis in Gansu Province.Methods With the method of stratified random sampling,all the administrative villages in pastoral areas,half pastoral areas,agriculture and urban areas in the 72 counties in Gansu Province were selected;according to the layers of the population of the county population proportion calculated,the layer investigation numbers were determined,and a total of 16 administrative villages were selected in each county.In the 16 administrative villages,more than 200 people of permanent residents were surveyed in each village (shortage was made up from a nearby village),at least 3 200 people were investigated in a county.By using the combined method of inquiry and field observation,natural factors,such as drinking water,farming and animal husbandry production data were surveyed.All data were analyzed using SPSS 19.0 statistical software.Unauy linear and multiple linear regression analysis of influencing factors and the relationship between the prevalence were analyzed.Results Unary linear regression analysis showed that longitudes,latitudes,altitudes,average annual temperature,annual rainfalls,secondary industries,animal husbandry towns,drinking water sources,the number of epidemic towns,epidemic villages,per capita net incomes in animal husbandry regions,the total number of livestock and the number of sheep were factors influencing the prevalence of echinococciasis (F =4.705,P ＜0.05).Latitude,altitude,animal husbandry towns,the number of epidemic towns,epidemic villages,per capita net incomes in animal husbandry regions,the total number of livestock and the number of sheep and the prevalence of echinococciasis was positively correlated (r =0.282,0.285,0.387,0.508,0.540,0.317,0.475,0.594,all P ＜0.05);longitude,average annual temperature,annual rainfall,secondary industries,drinking water sources and the prevalence of echinococciasis was negatively correlated (r =-0.311,-0.244,-0.244,-0.389,-0.311,all P ＜0.05).Multiple linear regression analysis showed that differences of interactions of annual rainfalls,drinking water sources,epidemic villages,per capita net incomes of herdsmen,secondary industries and the total number of livestock between groups were statistically significant (t =-1.822,-3.920,3.013,1.715,-1.609,3.264,all P ＜0.05).Conclusion The factors influencing the prevalence of echinococciasis in Gansu Province are correlated with annual rainfalls,drinking water sources,epidemic villages,per capita net incomes of herdsmen,secondary industries and the total number of livestock.

Objective:To describe the expression profiling of microRNAs of patients with non-small cell lung cancer. Methods:We reduced the scope of downregulated microRNAs in patients with non-small lung cancer by high throughput microarray based on quantitative real-time PCR (qRT-PCR). In this process, the downregulated microRNAs of non-small cell lung cancer tissues can be de-tected and compared with adjacent healthy lung tissues with relatively small samples. Different microRNA expression levels in non-small cell lung cancer and adjacent healthy lung tissues were verified in a large sample by RT-PCR. Results:A total of 20 types of microRNAs were significantly downregulated in non-small cell lung cancer tissues compared with adjacent healthy lung tissues. These microRNAs were listed as follows: hsa-miR-1; hsa-miR-22; hsa-miR-27a; hsa-miR-27b; hsa-miR-125a-5p; hsa-miR-125b;hsa-miR-142-5p;hsa-miR-143;hsa-miR-148a;hsa-miR-148b;hsa-miR-370;hsa-miR-373;hsa-miR-381;hsa-miR-489;hsa-miR-519a;hsa-miR-376c;hsa-miR-206;hsa-miR-380-5p;hsa-miR-223-5p;and hsa-miR-520c-3p. Among these microRNAs, 13 types were down-regulated in non-small cell lung cancer tissues as verified by RTPCR. These 13 types of microRNAs were listed as follows:hsa-miR-125a-5p; hsa-miR-143; hsa-miR-519a; hsa-miR-223-5p; hsa-miR-1; hsa-miR-520c-3p; hsa-miR-489; hsa-miR-27a;hsa-miR-373; hsa-miR-125b; hsa-miR-27b; hsa-miR-142-5p; and hsa-miR-206. Conclusion: In non-small lung cancer tissue, several kinds of microRNAs were significantly downregulated. Changes in microRNA expressions were significantly associated with tumori-genesis, progression, or other cancer cell functions in non-small cell lung cancer.

P210(bcr/abl) fusion gene is indispensable for generation and progression of chronic myeloid leukemia (CML). Small molecule inhibitors, such as imatinib, are effective for P210(bcr/abl) gene mediated CML, but drug resistance may occur. The unique fusion junction of P210(bcr/abl) gene is an attractive target for therapeutic intervention using RNA interference (RNAi). This study was purposed to constructed the BaF3 cell line by viral vector which can stably express P210(bcr/abl) shRNA and P210(bcr/abl) mRNA at the same time, and investigate the effect of lentiviral-victor-delivered shRNA on P210(bcr/abl) gene expression. The infective rate of lentiviral vector on BaF3 cells with P210(bcr/abl) gene was assayed by fluorescent microscopy; the cell proliferation ability was determined by trypan blue exclusion; the P210(bcr/abl) mRNA and protein expressions were detected by RT-PCR and Western blot respectively. The results found that stable expression of the P210(bcr/abl) shRNA resulted in obvious inhibition of P210(bcr/abl) mRNA and protein expression and increased sensitivity of these P210(bcr/abl) gene transformed Ba/F3 cells to imatinib. The IC(50) to imatinib in these cells decreased < 50% as compared with Ba/F3-P210(bcr/abl) cells which did not express P210(bcr/abl) mRNA. The survival time of the lethal dose irradiated mice induced by intravenous injection of these Ba/F3 cells was longer than the other group induced by Ba/F3-P210(bcr/abl). It is concluded that stable expression of shRNA targeting the P210(bcr/abl) gene fusion junction may potentiate the effects of conventional therapy for CML.
Animals ; Fusion Proteins, bcr-abl ; genetics ; metabolism ; Gene Expression ; Genetic Vectors ; Lentivirus ; genetics ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; metabolism ; Mice ; NIH 3T3 Cells ; RNA, Small Interfering ; genetics

This study reported the analysis of plasma phospholipid metabolism of the rats and the pathological biomarkers between the type 2 diabetes model control group (MC) and the normal control group (NC). SD rats were randomly divided into 2 groups: NC and MC. To investigate state of plasma metabolite profiling in normal body, type 2 diabetes mellitus (T2DM) model group using UPLC-Q-TOF/MS which was used as analysis tool in this research. The compounds were identified by UPLC-Q-TOF/MS based on MS/MS fragment ions information, element composition in MassLynx 4.1 and the Lipid Maps database. The sign of two groups of samples in specific markers for screening was through a software package in R software (BioMark software). The results show that the pathological markers were mainly phosphatidylcholine (PC) and triglycerides (TG); the 2-acyl PC in the MC group was less more obviously than that in the NC group; high carbon number and high degree of unsaturation of the TG was reduced under the condition of type 2 diabetes. In the state of type 2 diabetes, metabolic changes occurred in rat plasma phospholipids obviously, which had a close relationship with the occurrence and development of T2DM.
Animals ; Biomarkers ; blood ; Chromatography, High Pressure Liquid ; Diabetes Mellitus, Experimental ; blood ; Diabetes Mellitus, Type 2 ; blood ; Metabolome ; Phospholipids ; blood ; Rats ; Rats, Sprague-Dawley ; Tandem Mass Spectrometry

To establish vascular endothelial growth factor (VEGF) and interleukin-8 (IL-8) as secretary biomarkers for cell growth on topographic substrates, we have evaluated the secretion and expression of these 2 factors by SH-SY5Y human neuroblastoma cells on poly-L-lactide (PLLA) micropillar arrayed topographic substrates. We fabricated topographic substrates with UV lithography, silicon etching and polydimethylsiloxane-based replica molding, and interfaced SH-SY5Y human neuroblastoma cells with both the topographic substrates and PLLA flat substrates. Cell morphology and spreading were examined with scanning electron microscopy. The secretion and mRNA expression of VEGF and IL-8 were evaluated with enzyme linked immunosorbent assay (ELISA) and real time qPCR, respectively, 24 hours after cell plating. We successfully achieved 4 topographic substrates with a nominal pillar diameter of 2 microm and 4 microm, and a nominal pillar spacing of 2 microm and 7 microm. We found that the secretion and mRNA expression of VEGF and/or IL-8 by SH-SY5Y cells on 2-2 microm (pillar diameter-spacing), 4-2 microm and 4-7 microm topographic substrates were upregulated in comparison to those by cells on PLLA flat substrate, 24 hours after cell plating. Furthermore, both cytokines were even more substantially upregulated on the 2-7 microm substrate than on the other 3 topographic substrates. Compared to those on PLLA flat substrate, cells on topographic substrates showed significant changes in morphology (spreading area, perimeter and roundness), and the increase in the secretion and mRNA expression of VEGF and IL-8 was accompanied with a decrease in cell spreading areas. These results provided evidence that pillar arrayed topography was an important microenvironmental factor in affecting VEGF and IL-8 expression or secretion, and VEGF and IL-8 might serve as important secretary biomarkers for growth on topographic substrates by SH-SY5Y cells.
Biomarkers ; Cell Line ; Cell Proliferation ; Cellular Microenvironment ; Humans ; Interleukin-8 ; genetics ; secretion ; Neuroblastoma ; secretion ; Polyesters ; chemistry ; RNA, Messenger ; genetics ; Vascular Endothelial Growth Factor A ; genetics ; secretion

Objective To investigate the protective effect of rosiglitazone on the rats with high altitude pulmonary edema.Methods Thirty-six SD rats were randomly (random number) divided into 6 groups (n =6 each):control group (Control),hypobaric hypoxia model group (HH),rosiglitazone groups (RSG) which were administered with 3 different doses [RSG-L:5 mg/ (kg · d),RSG-M:10 mg/ (kg·d),RSG-H:20 mg/ (kg· d)],dexamethasone group [Dex,4 mg/ (kg· d)].Rats were injected intraperitoneally with different doses of rosiglitazone (RSG),dexamethasone (Dex) or vehicle (Control and HH) for 3 days before placed in simulated altitude of 6 000 m hypobaric hypoxia animal chamber where the temperature and pressure were constant.After 72 h in the chamber,each rat was anesthetized.The water content of lung was determined with wet/dry weight ratio.Bronchoalveolar lavage fluid was measured by bradford method.The contents of GSH was measured by micro-ezymed labeled method.The contents of MDA was measured by TBA method.The enzymatic activities of SOD was measured by WST-1 method.The changes of the TNF-α,IL-6 and IL-10 in serum were determined by ELISA.Light microscope was used to observe the pathological changes of lung tissue.Results Compared with Control group,the wet/dry weight ratio of lung (5.08 ± 0.24) and total protein content of BALF (351.06 ± 44.55) μg/mL increased significantly (P ＜ 0.01) in HH group.There were red blood cells in the alveolar and interstitium,pink fluid exudation in the alveolar,the alveolar septum enhancement,and a large number of inflammatory cell infiltration;the SOD activity (10.65 ± 0.94) U/mgprot and the content of GSH (1.63 ±0.20) μmol/gprot in lung tissue were significantly decreased (P ＜ 0.01),the contents of MDA (2.1 5 ± 0.18) nmol/mgprot increased significantly (P ＜ 0.01),TNF-o (56.92 ± 2.87) pg/mL and IL-6 (217.80 ±48.01) pg/mL levels in serum were significantly increased (P ＜0.01),and IL-10 (76.85 ± 16.72) pg/mL level decreased (P ＜ 0.05).Compared with the HH group,the wet/dry ratio of lung and total protein content of BALF in different doses of rosiglitazone group significantly decreased (P ＜ 0.01),the pathological changes of the lung tissue was significantly improved,SOD activity and the content of GSH in lung tissue was significantly increased (P ＜ 0.01),the content of MDA decreased (P ＜ 0.01),The levels of TNF-α and IL-6 in serum were significantly decreased (P ＜ 0.01),while the IL-10 level was significantly increased (P ＜ 0.01).Conclusion Rosiglitazone could protect the high altitude pulmonary edema by alleviating the oxidative stress and inflammatory response.

Objective To obtain the recombinant corticotropin releasing hormone (CRH) protein with soluble, high purity protein through optimizing prokaryotic expression condition and purifying glutathione thiol transferase (GST)-CRH protein. Methods To detect the expression of soluble CRH protein through grope of the host strain GST-CRH temperature of induction expression, the host strain concentration (OD600), IPTG concentration and induction time, the purification of GST-CRH was performed by GST-CRH agarose gel. Western Blot assay was used for the expression identification of the target protein. Results The optimal conditions for the induction of CRH protein were determined: temperature of 30 ℃, IPTG induced concentration 0.1 mmol/L, bacteria density (OD600) 0.8, the induction time of 8 hours, purified GST-CRH>95% fusion protein was obtained. Conclusion The optimal expression conditions of GST-CRH are obtained, and the soluble protein of high purity GST-CRH is also obtained.