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.

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.

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.

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

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.

Animals ; Animals, Newborn ; Brain ; blood supply ; metabolism ; pathology ; Carrier Proteins ; genetics ; DNA-(Apurinic or Apyrimidinic Site) Lyase ; genetics ; Gene Expression ; Hypoxia-Ischemia, Brain ; genetics ; pathology ; In Situ Hybridization ; Protein Tyrosine Phosphatase, Non-Receptor Type 13 ; Protein Tyrosine Phosphatases ; genetics ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Wistar

Objective To investigate the expression of hypoxia-inducible 1 alpha(HIF-lα)and glucose transporter 1(Glut1)in human breast cancer and its relationship to proliferating cell nuclear antigen (PCNA)protein and clinical pathologic factors.Methods Immunohistochemical staining was used to measure the expression of HIF-lα.Glut1 and PCNA in human breast fibroadenoma,usual hyperplasia and breast carcinoma.Results HIF-1α expression was not found in breast fibroadenoma and hyperplastic Iesions.In contrast.the positive rate of HIF-1α was found in the ductal carcinoma in situ 55%(DCIS,11/20)and the invasive breast carcinoma 85%(51/60).Glut1 positivity in breast carcinoma was 58.8%(47/80).The totsl positive rate of PCNA in breast carcinoma was 75%(60/80),that in DCIS was 65%(13/20)and that in invasive carcinoma was 78.3%(47/60).There was a positive correlation between HIF-lα and Glut 1 level (r=0.653,P＜0.01),a positive correlation between HIF-1α and PCNA level(r=0.693,P＜0.01);and also a positive correlation between Glutl and PCNA level(t=0.742.P＜0.01).conclusion The overexpression of HIF-lα and its target gene Glut1 played important roles in carcinogenesis and progression of breast carcinoma and closely correlated with cell proliferation of breast carcinoma and may become a new target for treatment of breast carcinoma.