Objective To evaluate the possibility and efficiency of nanoparticles as a new vector in vascular endothelial growth factor (VEGF) gene transfection. Methods Nanoparticle-VEGF (Np/VEGF)complex was prepared with poly (D, L-lactide-co-glycolide) (PLGA) loading VEGF165 gene using the multiple emulsion (w/o/w) technique. The envelopment efficiency and size of the complex were determined. Rat myocardial cells were cultured in vitro, and the Np/VEGF was transfected into the cultured myocardial cells. Then RT-PCR and ELISA were used to evaluate whether the Np/VEGF increased the level of gene expression. Four New Zealand rabbits were used, the suspension of Np/VEGF was injected into myocardial tissue of rabbits after thoracotomy. 96h after the operation, the tissue sections of the implant sites were observed with transmission electron microscope (TEM) to determine the process of nanoparticles as vectors for gene transfer to cardiac myocytes. Results The envelopment efficiency and size of the Np/VEGF complex thus prepared were 1.87% and 25-300nm respectively. RT-PCR and ELISA showed that VEGF gene could be successfully transfected into myocardial cells by nanoparticle, and NP/VEGF significantly enhanced gene transfection efficiency, and it was more effective than plasmid. 96h after the operation, a great number of nanoparticles were observed in myocardial cytoplasm and nucleus with TEM, and many nanoparticles began to dissolve and degrade, suggesting that the DNA was released slowly from the nanoparticles localized in the cytoplasmic compartment, and was then transferred into the nucleus. Conclusions NP/VEGF can act as a vector to transfect VEGF gene in vitro and in vivo, it significantly enhanced gene transfection efficiency, and it was more effective than plasmid.

Objective To construct a recombinant interfering RNA(miRNA)plasmid vector targeting annexinA3 and observe its impact on the growth and apoptosis of human gallbladder cancer cell line SGC-996.Methods Four small fragments of miRNA and one negative control were sequenced and cloned into vector pcDNATM 6.2-GW/EmGFPmiR.The recombinant plasmids were then transfected into gallbladder cancer cell line SGC-996 by positive liposomal transfection.The transcription and translation level of annexinA3 expression were detected by RT-PCR and Western Blot.Meanwhile, the proliferation and apoptosis of transfected tumor cells were determined by MTT and flow cytometry.Results The recombinant plasmids containing annexinA3 miRNA were successfully constructed.AnnexinA3 expression was significantly knocked down after miRNA plasmid transfection.Transfection of annexinA3 gene miRNA could effectively inhibit the proliferation of gallbladder cancer cell and promote apoptosis.Conclusion AnnexinA3 gene silenced by miRNA can induce the apoptosis of human gallbladder cancer cell line SGC-996.The process might serve as a potential approach for cancer gene therapy.

OBJECTIVE:To explore the regularity and characteristics of anaphylactoid reactions induced by Magnesium iso-glycyrrhizinate injection,and provide reference for clinical rational drug use. METHODS:27 ADR reports induced by Magnesium isoglycyrrhizinate injection from Hunan Center for ADR Monitoring from Jan. 2014 to Mar. 2015 were retrospectively analyzed. RE-SULTS:In the 27 cases,15 were male (55.56%) and 12 were female (44.44%),with percentage of 1.25∶1;ADR mainly oc-curred in patients with 41-60 years old(51.85%);involved organ/system were mainly skin and its appendages(35.85%)and sys-temic damage(26.42%),the main clinical manifestations were rash,itching,fever,chills,vomiting and dizziness;there were 7 severe ADR reports(25.92%),mainly showed chills,fever and other systemic symptoms;most ADR can be cured(40.74%)or relieved(55.56%)by related processing. CONCLUSIONS:There are no risk tips for severe allergic reactions in instruction,it is necessary to strengthen post-marketing surveillance and further improve the drug instructions to reduce the risk of severe ADR in-duced by Magnesium isoglycyrrhizinate injection.

Objective To observe the therapeutic effect of Professor Deng's Jinfu An Decoction(JAD)on intermediate and advanced non-small cell lung cancer(NSCLC).Methods Ninety patients with intermediate and advanced NSCLC were equally randomized into 3 groups.GroupⅠreceived routine chemical therapy(CT), groupⅡreceived oral use of JAD,which is mainly composed of raw Arisaema cum Bile,raw Rhizoma Pinelliae, Radix Pseudostellariae,Culmus Phragmitis,Poria,Semen Persicae,Bulbus Fritillariae Thunbergii,Gekko Chinensis,Pseudobulbus Cremastrae seu Pleiones,Radix Salviae Mihiorrhizae,and groupⅢreceived JAD + CT. Three weeks constituted one phase,two phases constituted one treatment courses,and the treatment lasted 2 courses.The changes of clinical symptom and signs scoring,tumor mass,immune function,hemorrheological indexes,and quality of life(QOL)as well as body weight and toxic and side reactions were observed in the three groups.Results The symptom scores decreased in the 3 groups obviously(P0.05).Between the 3 groups,the improvement of QOL in groupsⅡandⅢdiffered from that in groupⅠ(P

Objective To evaluate the outcomes of this reform to the hospital in the past two years.Methods Collecting such indexes as expenses composition,service volume and efficiency of the year before,the first year and the second year of the reform made at the hospital.Comparing the statistics and analyzing the effectiveness.Results Average outpatient expenses per visit and outpatient drug costs of patients with medical insurance during the two years of reform were found to be 12.03% and 5.7%respectively lower than the year before;while the average expenses per case fell by 4.08% and 2.69%respectively,with drug costs falling significantly as well.Outpatient visits fell,with steady increase of outpatients with medical insurance coverage.The distribution of outpatient visits varies significantly in terms of medical branches.There are a slight surplus of medical service charges of outpatients,and slight deficit of medical service charge of inpatients,with a smooth shift of the income makeup from dependence on drug price markup to medical service charge,and steady improvement of hospital efficiency.Conclusion The reform can alleviate patients’financial burden,encourage rational visits of doctors,promote fine management and structure adjustment,benefiting patients in the end.

Breast Neoplasms ; therapy ; Consensus ; Female ; Humans ; Neoadjuvant Therapy

Objective　To study the value of B type ultrasonography guiding percutaneous radiofrequency ablation (PRFA) for hepatic cancer (HC). Methods　70 patients with primary or metastatic hepatic malignancies were treated by PRFA under ultrasound guidance through LeVeen multipolar array needle electrode and RF 2000. All patients were followed up by turnor marker, B type sounography and MRI monthly. Results　PRFA were performed in 53 patients with hepatocellular carcinoma (HCC) and 17 cases with metastatic liver cancer. Liver function classfication was 53 cases in Child class A, 15 in class B and 2 in class C. In 21 patients with non-operated small HCC (≤5 cm), 12 of 17 patients with AFP positive were decreased to normal (70.6%) and decreased markedly in 4 (23.5%), and MRI showed the treated tumors were completely necrosis in 16 patients (76.2%). The 3, 6 month survival rates were 90.5%, 100% respectively in small HC groups, and 66.7% to 72.7% and 27.6% to 72.7% in large HC groups.Conclusions　PRFA as a local thermal therapy is becoming a kind of new palliative treatment for hepatic malignancies, which not only is safe, minimally invasive, and effective for small ones; but also can be used with combination of TACE for larger ones.

The object of this study is to preparate the berberine hydrochloride (BBH) resin compound with taste masking effect. We took the BBH as the model drug and Amberlite IRP69 as the drug carriers, uncovered the curve of solubility of BBH in different cosolvent with a certain range of temperature, and then used it to calculate the parameters during the preparation of the complex such as adding quantity of BBH and the reaction temperature. Afterwards, the characteristic and in vitro release experiments were studied to verify the formation and predict the in vivo release behavior of the complex. The results showed that in the condition of using 60% ethanol as a cosolvent and stirring at 50 ℃ for 1 h, the drug loading and drug availability of the complex are at about 35% and 64%, respectively, and has a better taste-masking effect. In this study, a method was provided for preparing a taste-masking preparation of BBH.

Objective To investigate the mechanism of acute inflammatory response and tissue repair when rats accepted transplanted bone marrow mesenchymal stem cells (MSC) in severe acute pancreatitis (SAP).Methods A total of 40 rats were randomly divided into 4 groups which included the normal group (n=10),the model severe acute pancreatitis group (n=10),the transplanted bone marrow mesenchymal stem cells group (n =10),and the combination of bone marrow mesenchymal stem cells and granulocyte colony-stimulating factor (G-CSF) group (n=10).To cure the acute severe pancreatic injury caused by SAP,rats were injected with EDU-labeled MSCs and granulocyte colonystimulating factor (Gr-CSF).The conversion rate of MSCs to pancreatic cells or MSCs to endothelial cells was detected to assess the role of MSCs in tissue regeneration and repair.The expression of amylase,interleukin-6 (IL-6),and interleukin-10 (IL-10) in serum was detected to assess the role of MSCs in an acute inflammatory response.Results The concentrations of amylase and IL-6 were reduced and the concentration of IL-10 was increased in MSC and MSC+G-CSF groups after the onset of SAP.Flow cytometry showed a small amount of MSCs converting to endothelial or pancreatic cells,but the conversion rate was relatively low.Conclusions MSCs can play an important role in the antipre-release of inflammatory mediators,reducing acute immune response to control the acute inflammatory response in SAP.Moreover,MSCs can repair a damaged pancreas by converting into both pancreatic and endothelial cells.

Objective To investigate effects of α-crystallin on proliferation of lipopolysaccharide (LPS)-activated retinal microglia and bioactivity of iNOS.Methods The retinal microglial cells cultured in vitro were analyzed and their purity was identified by cell immunofluorescence and flow cytometry.After microglia cells being intervened using LPS and α-crystallin at various concentrations,influence of α-crystallin on activity of LPS-activated retinal microglia was detected by MTT method and level of NO was measured by RT-PCR to observe changes of iNOS expression in microglia.Results Purity of primary cultured microglial cells was 94.15％ by GSA-IB4 immunohistochemical identification and 93.34％ by CD11b flow cytometry.α-crystallin of 10-4g/L awakened activity of microglia induced by 10-6g/L LPS (P ＜ 0.01).Expressions of iNOS protein and mRNA showed significant decrease in combined treatment group (P ＜ 0.05).Conclusion In clinical condition,α-crystallin decreases the harm of microglial cells on retinal ganglial cells (RGCs) after optical nerve injury by inhibiting the microglia cells to produce NO and iNOS.