Objective To study the effects of pcDNA3/AFP/TK/Angio fusion gene targeting therapy for human primary liver cancer in nude mice implanted with SMMC-7721. Methods Human liver cancer cell line SMMC-7721 was implanted subcutaneously in nude mice to establish experiment model. Animals bearing liver cancer were randomly divided into five groups: control group， vector group， GCV (ganciclovir) group， pcDNA3/TK/Angio group; pcDNA3/AFP/TK/Angio group. Different plasmids were directly injected into tumors and GCV was intraperitoneally administrated simultaneously according to different groups. The growth of tumors was observed and the pathology was examined as well. Serum AFP level was measured by radioimmunology, the ultrastructural change of tumor cells was studied by using electron microscopy, the expressions of MVD and VEGF were respectively detected with immunohistochemistry and the cell apoptosis in situ was detected by TUNEL. Results The success rate to establish subcutaneous implanted liver cancer model in nude mice was 100%. The tumor volume, serum AFP level, VEGF and MVD expressions of pcDNA3/TK/Angio group and pcDNA3/AFP/TK/Angio group were lower than those in control group, vector group and GCV group (P＜0.05) and more apoptosis cells could be observed. While the tumor volume, serum AFP level, VEGF and MVD expressions of pcDNA3/AFP/TK/Angio group was lower than those in pcDNA3/TK/Angio group (P＜0.05); and apoptosis index was higher than that of the latter (P＜0.05).Conclusion pcDNA3/AFP/TK/Angio fusion gene inhibits the growth of tumor remarkably and becomes a promising new biological agent to treat human primary liver cancer.
Objective To systematically review the health economic evaluation studies of medicines for the treatment of acute myeloid leukemia (AML). MethodsThe PubMed, EMbase, Cochrane Library, CBM, CNKI, and WanFang Data, as well as the CRD database specifically for health economics were electronically searched from inception to June 2022, and related journals in the field of health economics and the websites of HTA institutions in various countries were manually searched. The quality of the studies was assessed using the CHEERS checklist. The basic characteristics of health economics evaluation publications were summarized, the quality of model structures and methodologies was assessed and economic evaluation results were compared among different treatments. Results A total of 17 studies were included, and cost-effectiveness analyses were conducted from the perspectives of the health system, patients, the whole society, and medical insurance payers. The economic evaluation models were relatively unified, but there were differences in methods and results reporting, and the quality needed to be improved. The research objects were mainly the comparison of hypomethylating agents, targeted medicine and traditional chemotherapy regimens, as well as the comparison of different chemotherapy combinations and different drug dosages. Conclusion Real-world studies are mainly focused on traditional chemotherapy regimens, and model-based health economic evaluations, such as Markov models, are more frequently applied to newly developed targeted drugs and demethylation drugs. Among all treatments, the chemotherapy regimens including cytarabine, midostaurin, and decitabine are found to be more cost-effective.
ObjectiveTo analyze targeted surveillance results of nosocomial infection in Neurosurgical Intensive Care Unit (ICU) and investigate the characteristics of nosocomial infection, in order to provide reference for constituting the intervention measures. MethodsWe monitored the incidence of nosocomial infection, the application and catheter-related infection of invasive operation, and the situation of multiple resistant bacteria screening and drug resistance characteristics of each patient who stayed more than two days in neurosurgical ICU during January to December 2013. ResultsThere were a total of 1 178 patients, and the total ICU stay was 4 144 days. The nosocomial infection rate was 4.92%, and the day incidence of nosocomial infection was 13.75‰. The nosocomial infection rate was significantly higher in January and between July and December compared with other months. Ventilator utilization rate was 9.75%; ventilator-associated pneumonia incidence density was 14.85 per 1 000 catheter-days; central line utilization rate was 28.40%; central line-associated bloodstream infection incidence density was 0.85 per 1 000 catheter-days; urinary catheter utilization rate was 97.90%; and the incidence density of catheter-associated urinary tract infection was 0.25 per 1 000 catheter-days. ConclusionThe nosocomial infection rate has an obvious seasonal characteristic in neurosurgical intensive care unit, so it is necessary to make sure that the hospital infection control full-time and part-time staff should be on alert, issue timely risk warning, and strengthen the risk management of hospital infection.
ObjectiveTo investigate the microRNA (miRNA) expression profile during chondrogenic differentiation of human adipose-derived stem cells (hADSCs), and assess the roles of involved miRNAs during chondrogenesis. MethodshADSCs were harvested and cultured from donors who underwent elective liposuction or other abdominal surgery. When the cells were passaged to P3, chondrogenic induction medium was used for chondrogenic differentiation. The morphology of the cells was observed by inverted phase contrast microscopy. Alcian blue staining was carried out at 21 days after induction to access the chondrogenic status. The expressions of chondrogenic proteins were detected by ELISA at 0, 7, 14, and 21 days. The miRNA expression profiles at pre- and post-chondrogenic induction were obtained by microarray assay, and differentially expressed miRNAs were verified by real-time quantitative PCR (qRT-PCR). The targets of the miRNAs were predicted by online software programs. ResultshADSCs were cultured successfully and induced with chondrogenic medium. At 21 days after chondrogenic induction, the cells were stained positively for alcian blue staining. At 7, 14, and 21 days after chondrogenic induction, the levels of collogen type Ⅱ, Col2a1, aggrecan, Col10a1, and chondroitin sulfate in induced hADSCs were significantly higher than those in noninduced hADSCs (P<0.05). Eleven differentially expressed miRNAs were found, including seven up-regulated and four down-regulated. Predicted target genes of the differentially expressed miRNAs were based on the overlap from three public prediction algorithms, with the known functions of regulating chondrogenic differentiation of stem cells, selfrenewal, signal transduction, intracellular signaling cascade, and cell cycle control. ConclusionA group of miRNAs and their target genes are identified, which may play important roles in regulating chondrogenic differentiation of hADSCs. These results will facilitate the initial understanding of the molecular mechanism of chondrogenic differentiation in hADSCs and subsequently control hADSCs differentiation, and provide high performance seed cells for cartilage tissue engineering.
ObjectiveTo construct tumor specific tubercle bacillus antigen Ag85A gene lentiviral vector driven by murine telomerase catalytic subunit promoter (PmTERT), paving the way for further research in tumor targeting immuno-gene therapy. MethodsPmTERT was amplified by PCR method, with murine genomic DNA as template. Then, transcriptional activities of PmTERT in various murine and human cell strains were studied by luciferase assay. Ag85A expression lentiviral vectors driven by cytomegalo virus (CMV) promoter and PmTERT respectively (pLVX-Ag85ACMV and pLVX-Ag85A-PmTERT) were constructed with nucleic acid cloning approach. And above recombinants were verified with DNA sequencing and Western blot. ResultsLucifease assay revealed that 331 bp PmTERT cloned in present research had transcriptional activity in murine Lewis lung cancer cells, human lung adenocarcinoma cells A549, and human esophageal cancer cells EC-109, while no transcriptional activity in murine fibroblasts NIH3T3 and human embryo fibroblasts MRC-5. Western blot revealed expression of Ag85A in pLVX-Ag85A-CMV transfected Lewis and NIH3T3 cells, pLVX-Ag85A-PmTERT transfected Lewis cells, no expression in pLVX-Ag85A-PmTERT transfected NIH3T3 cells. ConclusionPmTERT has tumor specific transcriptional activity. Ag85A gene can express selectively in tumor cells, driven by PmTERT.
Objective To construct the recombinant of hepatocellular carcinoma-targeting adenovirus containing r-Caspase-3 gene and provide the gene therapic strategy for hepatocellular carcinoma. Methods The pAdTrack-EAFP-PALB was constructed and the r-Caspase-3 gene was subcloned into the vector. The linearized shuttle plasmid was homogenously recombined with AdEasy-1 in BJ5183 cells. The candidate clone was analyzed by restriction endonuclease digestion and sequencing, and then pAdEasy-EAFP-PALB/r-Caspase-3 vector was digested with PacⅠand transfected into AD293 cells for packaging and amplifying, recombinant virus was constructed successfully. Infection titer and efficiency of recombinant virus were monitored by green fluorescent protein (GFP) expression. The expression of r-Caspase-3 in infected HepG2 cells was detected by RT-PCR and Western blot. The apoptosis of HepG2 cells was detected by SRB dyeing method. Results Shuttle vector pAdTrack-EAFP-PALB/r-Caspase-3 was correct after identification by restriction endonuclease analysis and sequencing. By PCR and PacⅠ restriction endonuclease analysis， the homologous recombinant of pAdEasy-EAFP-PALB/r-Caspase-3 was successful. The expression of GFP was observed when linearized pAdEasy-EAFP-PALB/r-Caspase-3 was transfected into AD293 cells. AD293 cells could be infected repeatedly by recombinant adenovirus. The expression of r-Caspase-3 gene on HepG2 cells was detected by RT-PCR and Western-blot methods respectively, which confirmed that the Ad-EAFP-PALB/r-Caspase-3 was constructed successfully. The specificity of Ad-EAFP-PALB/r-caspase-3 which targeting induced hepatocellular carcinoma cells was founded by SRB dyeing test. Conclusion The Recombinant of hepatocellular carcinoma-targeting adenovirus containing r-Caspase-3 gene was constructed successfully and which established the foundation of r-Caspase-3 gene therapy in future research to hepatocellular carcinoma.
Objective To investigate nosocomial infection rate in Intensive Care Unit (ICU), its risk factors and the pathogenic characteristics of multidrug-resistant bacteria through targeted monitoring, in order to provide scientific references for reducing nosocomial infection. Methods Targeted monitoring was performed on the patients who were admitted to the comprehensive ICU between July 2014 and June 2016. Results Nosocomial infection occurred in 312 of the 4 991 patients. The case infection rate was 6.25%, and case infection rate per day was 19.03‰. After the adjustment, the case infection rate per day was 6.77‰. The ventilator-associated pneumonia infection accounted for 30.78‰; catheter-related bloodstream infection occupied 0.30‰; and catheter-associated urinary tract infection accounted for 0.27‰. The respiratory tract was the major part of nosocomial infection, accounting for 90.38%. Gram-negative bacilli were the major bacteria accounting for 92.74%, in whichAcinetobacter baumannii accounted for 36.29%. Conclusions Through targeted monitoring to keep abreast of the current situation of nosocomial infection in ICU, management and interventions can be targeted. It is an important way to reduce nosocomial infection in ICU.
Objective To evaluate the efficacy and safety of anti-vascular endothelial growth factor (VEGF) agents for advanced renal cell carcinoma. Methods We searched MEDLINE, EMbase, The Cochrane Library, CBMdisc and China Academic Periodical database from the establishment of each database to April 2009. We included randomized controlled trials (RCTs) that evaluated anti-VEGF agents (sunitinib, sorafenib and bevacizumab). The quality of the included trials was evaluated by two reviewers independently. Meta-analyses were conducted by the Cochrane Collaboration’s RevMan 4.2 software. Results Four RCTs involving 2 320 patients were identified. According to the different interventions for advanced renal cell carcinoma, we divided the patients into two groups: anti-VEGF agents monotherapy and anti-VEGF agents plus interferon combination treatment. Our meta-analyses showed: monotherapy was superior to interferon on inhibition of tumor progression [OR=0.38, 95%CI (0.29, 0.51), Plt;0.01] and control of tumor [OR=2.53, 95%CI (1.87, 3.43), Plt;0.01], but was not significantly different from interferon on the overall effective rate [OR=1.97, 95%CI (0.20, 19.57), P=0.56] and serious side effects [OR=1.98, 95%CI (0.90, 4.34), P=0.09]. There were significant differences between anti-VEGF agents plus interferon and interferon alone on inhibition of tumor progression [OR=0.67, 95%CI (0.53, 0.84), P=0.000 5], overall effective rate [OR=2.65, 95%CI (1.94, 3.61), Plt;0.01], control of tumor [OR=2.14, 95%CI (1.65, 2.78), Plt;0.01] and serious side effects [OR=2.63, 95%CI (2.09, 3.31), Plt;0.01]. Conclusion Compared with interferon, anti-VEGF agents could inhibit tumor progression more effectively. Moreover, the combination therapy with interferon could offer a more favorable overall effective rate for advanced renal cell carcinoma, but then followed by more serious side effects. We need to weigh the merits and demerits of drugs before making a clinical decision for advanced renal cell carcinoma.
ObjectiveTo reveal the true value of plasma detection of epidermal growth factor receptor (EGFR) mutation for early-stage non-small cell lung cancer (NSCLC) gene diagnosis and to predict survival prognosis. MethodsTissue samples of positive EGFR mutations by using amplification refractory mutation system (ARMS) method were surgically resected from 198 patients with stage I-IV NSCLC between February 2014 and June 2015 in Tangdu hospital. Paired blood samples were collected before surgery. And the cellfree DNA (cfDNA) in plasma was extracted, plasma EGFR mutations were detected by real-time polymerase chain reaction (PCR). Concentration of cfDNA was measured by ultraviolet spectrophotometry. Follow-up observation for stage ⅢA patients was put into force after surgery. Kaplan-Meire was used in survival analysis. ResultsThe sensitivity of EGFR mutation for the 198 paired tissues and plasma samples was 17.2%.The sensitivity was positively correlated with TNM stage and negatively correlated with tumor differentiation. The sensitivity of sage ⅢA was 33.3%, significantly higher than that of the patients at stage ⅠA (1.6%, P=0.000) and stage ⅠB (7.9%, P=0.004). The sensitivity of poor differentiation was 36.8%, significantly higher than that of high differentiation (0.0%, P=0.000) and moderate differentiation (15.7%, P=0.010). There was no correlation between plasma cfDNA concentration and patient characteristics. Survival analysis showed that plasma detection was a vital factor for predicting postoperative survival prognosis of stage ⅢA patients (P=0.014). ConclusionTissue samples cannot be replaced by plasma samples for epidermal growth factor receptor (EGFR) mutation test in early-stage NSCLC patients, currently. When the sensitivity increases dramatically in the plasma samples of stage ⅢA NSCLC and poor differentiation tumor, we recommend using plasma detection for gene diagnosis, dynamic monitoring of EGFR mutations in stage ⅢA or poorly differentiated tumors, especially in NSCLC patients whose tissue samples cannot be obtained by surgery. And plasma EGFR detection is a valuable method of forecasting survival prognosis for locally advanced NSCLC patients.
Objective To review the advances of target gene therapy of liver cancer. MethodsWe analyze and compare the tissuespecific carrier system or cellspecific gene expressing system from current researches of liver cancer gene therapy. ResultsArtificial synthetic DNA transfer system and modified viral vectors could efficiently transfect target cells and get highlevel expression. The ciselements of alpha fetal protein or albumin gene have been often adopted in the regulation of therapeutic gene and have shown their effectiveness. Some other gene therapy strategies also promised a good future. Conclusion Searching for more specific and universal liver cancer antigens is the key to improve the target gene therapy efficiency. The individual situation is the basis to select the best transfer system or regulatory elements in the future.