OBJECTIVE:To study the antihepatocarcinoma activity and toxicity of Aclacimomycin-A solid lipids nanoparticle for injection(ACM-SLN)in nude mice in vivo with ACM for injection as the reference preparation.METHODS:The nude mice were divided into control group,ACM group and ACM-SLN group after tumor transplantation,which were injected with the corresponding medicine before the tumor-inhibition rates of which were calculated,which were then injected with ACM and ACM-SLN,respectively before the LD50 of mice were calculated.RESULTS:Compared with the control group,the tumor inhibition rates of ACM-SLN group and ACM group were 78.4%and 38.8%,respectively,the LD50 of which were 16.3 mg/kg and 18.0mg/kg,respectively.CONCLUSION:The ACM-SLN is superior to ACM in terms of the anti-tumor effect while without the increase of toxicity.

OBJECTIVE:To establish a RP-HPLC method for content determination of costunolide in Liqifuwei oral liquid.METHODS:The assay was performed on a Luna C 18 column by UV detector at the wavelength of225nm with methanol-water(72∶28)as the mobile phase at the flow rate of1.0ml/min.RESULTS:The amount of costunolide was linear with its area over the range from0.2?g～2.0?g(r=0.9996),the average recovery was98.4%(RSD=0.85%).CONCLUSION:The present method is convenient,sensitive and accurate with good reproducibility and can be used for the quality control of Liqifuwei oral liquid.

Objective To explore the causes of misdiagnosis of lung abscess and its clinical pathological characters. Methods The clinical data of autopsy-proven lung abscess in Beijing Hospital from 1980 to 1995 were reviewed. Results Twelve patients consisted of 11 males and 1 female. Their age ranged from 60 to 88 years. None of them was clinically diagnosed lung abscess before death. Concomitant dieases were severe and complex, mostly coronary disease, hypertension, COPD, cerebrovascular disease and diabetes mellitus. The clinical presentation and chest X-ray of lung abscess in the elderly were deceptively nonspecific and variable. Over 2 types of pathogens were isolated from sputum culture, mostly klebsiella pneumoniae, pseudomonas aeruginosa, candida. The autopsy showed 3 cases with a isolated lung abscess and 9 cases with lung multi-abscess. Conclusions The reasons for misdiagnosis of lung abscess in the elderly might be the variability of clinical presentation, the other concomitant disease. Frequent pursuit of chest CT scan in suspected cases is guarranted .

Objective:To understand the current situation and problems of the supervision of individual clinics in China , and put forward some reform suggestions .Methods:Two cities were selected from typical provinces in east-ern, central and western regions by typical sampling .The investigation was conducted by semi-structured interviews and typical clinic participant observation method .Results:There were serious problems in the regulation of the pri-vate clinics and it was necessary to build efficient regulation mechanism .Conclusions:We should strengthen the su-pervision of private clinics .In the future , we should improve the access threshold for the private clinic; strengthen inter-sector cooperation and joint law enforcement; promote information exchange and information network construc-tion;use economic incentives and punitive measures at the same time and make the association itself and social su -pervision work .

OBJECTIVE:To develop a RP-HPLC method for the determination of salicylic acid in serum and to apply this method to the pharmacokinetic and bioavailability study of salycylic acid in compound aspirin preparations.METHODS:Waters2690HPLC instrument was used with Diamonsil C 18 column(5?m,250mm?4.6mm)as stationary phase and methanol-ace?tonitrile-0.2%phosphoric acid(18∶32∶50)as mobile phase at a flow rate of1.0ml/min,and the detective wavelength was237nm.RESULTS:Calibration curve of salicylic acid was linear in the range of0.40～101.00?g/ml(r=0.9999).CONCLUS_ ION:The method is simple,sensitive,rapid and suitable for pharmacokinetic and bioavailability study of salicylic acid.

OBJECTIVE:To determine the concentration of valproic acid in serum.METHODS:Determination was performed with RP-HPLC with methanol:water(70∶30) as mobile phase,?-bromoacetophenone as deriving agent and cyclohexanecarboxylic acid as internal standard,and detected at wavelength 248nm.RESULTS:The calibrating curve of valproic acid was linear in the range of 14.47～248.0?g/ml.CONCLUSION:The method was convenient,rapid,accurate and suitable for TDM.

OBJECTIVE:To establish a GC-MS assay for determination of the contents of phenol and menthol in calamine and menthol lotion.METHODS:Using DM-5elastic quartz capillary as separation column,I-octanol as internal standard,phenol and menthol were detected separately under the70℃～150℃step-up hyperthermic condition to select the ion fragment peaks with M/Z of94and71.RESULTS:Phenol and menthol were good linear within1.092～21.840?g/ml(r=0.9998)and1.194～9.552?g/ml(r=0.9999).Recoveries were100.2%(RSD=1.34%)and100.4%(RSD=0.74%)respective?ly.CONCLUSION:The method is quick and accurate with highly repeatability and specificity and it is adequate for contents determination and quality control for this preparation.

Bovine interferon-gamma (BoIFN-gamma) gene was amplified by reverse transcription polymerase chain reaction (RT-PCR) from total RNA of bovine spleen lymphocytes stimulated with ConA. The products of RT-PCR were cloned into pVAX1 vector, positive recombinant clone was identified by restriction enzyme digestion and sequencing. The recombinant plasmid pVAX1-BolFN-gamma was transfected into COS-7 cells mediated by lipofectine, indirect immunofluorescent assay analysis confirmed that rBoIFN-gamma was expressed in COS-7 cells. BoIFN-gamma gene (without signal peptide) was cloned into pET-30a(+) and pGEX-6p-1 vector, and transformed into the Escherichia coli cells. After optimizing the induction condition, SDS-PAGE analysis showed that the expression products were all found in soluble form and had a molecular weight of 23 kDa and 43 kDa respectively. BoLFN-gamma precursor gene (with signal peptide) was cloned into transfer vector pFastBac 1, and transformated into DH10Bac E. coli cells. By site-specific transposition, BoIFN-gamma gene was integrated into shuttle vector Bacmid, and transfected into the Sf9 insect cells mediated by lipofectine to produce recombinant baculovirus. Indirect immunofluorescent assay analysis confirmed that rBac-BoLFN-gamma was expressed successfully in Baculovirus vector system. The antiviral activities of rHis-BoIFN-gamma, rGST-BoIFN-gamma and rBac-BoIFN-gamma were up to 8.389 x10(7) U/mg, 6.554 x10(5) U/mg and 4.096 x 10(4) U/mL respectively, which were analyzed in MDBK/VSV system. A sandwich ELISA was established using monoclonal antibodies 3E6 and 5G4, which can detect BoIFN-gamma in quantity and provide a useful method for the clinical practice and research of BolFN-gamma.
Animals ; Antiviral Agents ; pharmacology ; Baculoviridae ; genetics ; metabolism ; COS Cells ; Cattle ; Cercopithecus aethiops ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; genetics ; Interferon-gamma ; biosynthesis ; genetics ; pharmacology ; Recombinant Proteins ; biosynthesis ; genetics ; pharmacology ; Transfection

Objective:To investigate the effect of miRNA-296-5p (miR-296-5p) on the migration and invasion of hypoxia-induced pancreatic cancer cells and its related mechanisms.Methods:Human pancreatic cancer cell line PANC-1 was selected. Pancreatic cancer tissues from 55 pancreatic cancer patients who underwent the resection and adjacent carcinoma normal pancreatic tissues from 10 patients at Shanghai Fengxian District Central Hospital and Bengbu Medical College First Affiliated Hospital between January 2010 and December 2014 were collected. The expression levels of hypoxia-inducible factor 1α (HIF-1α) and miR-296-5p in tissue microarray of pancreatic cancer and adjacent carcinoma normal pancreatic tissues were detected by using immunohistochemistry and in situ hybridization. The relationship between miR-296-5p and HIF-1α as well as their correlation with clinicopathological characteristics of patients were analyzed. PANC-1 cells were divided into hypoxic group and normoxic group. Transwell assay was used to detect the cell migration and invasion ability of both groups. Real-time quantitative polymerase chain reaction (qRT-PCR) was used to examine the expressions of HIF-1α and miR-296-5p under hypoxic environment of both groups. The expression of HIF-1α was interfered by transfecting small interfering RNA (siRNA). PANC-1 cells were divided into PANC-1 group (the empty control), PANC-1-NC group (the negative control) and PANC-1-siRNA group. The expression of miR-296-5p was measured. After co-transfecting miR-296-5p agonist and miR-296-5p inhibitor, the cells were divided into Agomir-miR-296-5p group (agonist group), Agomir-miR-296-5p-NC group (agonist negative control group), Antagomir-miR-296-5p group (inhibitor group) and Antagomir-miR-296-5p-NC group (inhibitor negative control group). Transwell assay was used to detect the cell migration and invasion ability of all groups. Luciferase reporter gene system was used to verify whether miR-296-5p promoter region had binding site of HIF-1α.Results:The high expression rate of HIF-1α in pancreatic cancer tissues was higher than that of adjacent carcinoma normal pancreatic tissues [81.8% (45/55) vs. 0 (0/10), P<0.01], and the high expression rate of miR-296-5p in pancreatic cancer tissues was lower than that of adjacent carcinoma normal pancreatic tissues [12.7% (7/55) vs. 90.0% (9/10), χ2 = 27.23, P<0.01]. The expression of HIF-1α was negatively correlated with that of miR-296-5p ( r = -0.53, P<0.01). The low expression of miR-296-5p was closely related with the tumor diameter, TNM staging, lymph node metastasis (all P<0.05). The number of PANC-1 invasion cell was 15.3±2.1 in normoxic group and 24.7±1.5 in hypoxic group, and the difference was statistically significant ( t = 0.26, P = 0.003). The number of PANC-1 migration cell was 20.7±3.8 in hypoxic group and 32.7±1.2 in normoxic group, and the difference was statistically significant ( t = 5.25, P = 0.006). The relative expression level of HIF-1α mRNA in PANC-1 cell of hypoxic group was higher than that of normoxic group [(1.00±0.01) vs. (0.30±0.02)], and the difference was statistically significant ( t = 56.45, P<0.01); the relative expression level of miR-296-5p in PANC-1 cell of hypoxic group was lower than that of normoxic group [(1.14±0.04) vs. (3.05±0.20)], and the difference was statistically significant ( t = 16.05, P<0.01). The number of invasion cells in PANC-1 group, PANC-1-NC group and PANC-1-siRNA group was 24.7±1.5, 25.7±1.5, 12.0±1.7, respectively, and the difference was statistically significant ( F = 68.13, P<0.01).The cell invasion ability in PANC-1-siRNA group was decreased compared with that in PANC-1 group ( t = 9.50, P = 0.001). The number of cell migration was 32.7±1.2, 37±1.0, 17.3±1.2, respectively in PANC-1 group, PANC-1-NC group and PANC-1-siRNA group, and the difference was statistically significant ( F = 262.09, P<0.01). The cell migration ability in PANC-1-siRNA group was decreased compared with that in PANC-1 group ( t = 16.26, P<0.01). The cell invasion and migration ability in Antagomir-miR-296-5p group was increased compared with that in PANC-1 group (all P<0.05); the cell invasion and migration ability in Agomir-miR-296-5p group was decreased compared with that in PANC-1 group (all P<0.05). The results of luciferase activity detected by luciferase reporter gene system showed that miR-296-5p had the target binding to HIF-1α. Conclusions:HIF-1α plays a key role in the invasion and migration of hypoxia-induced pancreatic cancer cells through negatively reducing miR-296-5p.

Objective To investigate the effect of Guizhi Fuling Capsule on NF-κB and MAPK signaling pathway in lipopolysaccharide(LPS)-induced rat endometritis model.Methods The rats were randomly divided into 6 groups according to body weight,namely sham-operation group,model group,positive group(dexamethasone,2.5 mg·kg-1),Guizhi Fuling Capsule high-dose,medium-dose and low-dose groups(0.54,0.27,0.14 g·kg-1),with 10 rats in each group.The sham operation group and the model group were given 0.5%CMC-Na by gavage,and the other groups were given corresponding drugs by gavage once a day for 7 consecutive days.One hour after the last administration,the animals were anesthetized by intraperitoneal injection of 10%chloral hydrate.The rats in the sham operation group only underwent laparotomy and abdominal closure.The left uterus of the rats in the other groups was scratched and injected with a syringe(1.0 μg·μL-1)LPS normal saline solution 0.25 mL.24 hours after LPS injection,the uterine tissues of rats were collected and the pathological changes and MPO,TNF-α,IL-1β,IL-6 were measured in uterine tissues.The expression levels of NF-κB p65,IκBα,ERK,p38 and their phosphorylated proteins were measured in uterine tissues.Results Compared with sham operation group,histopathological score,MPO,TNF-α,IL-1β and IL-6 levels in model group were significantly increased(P<0.01),and NF-κBp65,IκBα,ERK,p38 protein phosphorylation levels were significantly increased(P<0.01).Compared with model group,Guizhi Fuling capsule significantly decreased pathological score of uterus,TNF-α,IL-1β,and IL-6 levels(P<0.01),and significantly decreased NF-κBp65,IκBα,ERK,p38 protein phosphorylation levels(P<0.01).Conclusion Guizhi Fuling capsule plays an anti-inflammatory role in endometritis by inhibiting the transmission of NF-κB and MAPK signaling pathways and inhibiting the secretion of inflammatory factors IL-1β,TNF-α and IL-6 in uterine tissue.