Objective To detection the methylation of p16INK4A in primary hepatocellular carcinoma, a nested bisulfite sequencing and methylation-specific polymerase chain reaction (BS-MSP) protocol was designed and used.Methods Bisulfite-modified DNA were amplified to evaluate the quality of templates with a pair of bisulfite sequencing primers in the first round of PCR, then subjected to methylation assay with corresponding methylation or unmethylation specific PCR primers.Representative PCR products were sequenced to confirm its correctness.Results 3 of 40 cases (7.5%) were failed to assay due to poor quality of templates, and 29 of 37 cases (78%) were detected p16INK4A methylation.Sequencing results confirmed that templates were correctly amplified.Conclusion BS-MSP technique might be valuable for methylation study on carcinogenesis and clinical assay.

OBJECTIVETo establish a quantitative technique for assaying gene methylation in hepatocellular carcinoma (HCC) and evaluate its feasibility for clinical application.

METHODSFollowing bisulfite modification and PCR amplification, the fragments of CDKN2A and ACTB were cloned into plasmids to generate calibration curves using SYBR Green quantitative PCR, and then these two genes were quantitatively analyzed in 41 cases of HCC specimen.

RESULTSThe amplification curve, dissociation curve, calibration curve and electrophoresis analysis showed that SYBR Green fluorescent quantitative PCR could assay 10(2)-10(8) copies/microL of recombinant plasmids with high specificity, high sensitivity and a wide detection range. The tests on 41 cases of HCC specimens further confirmed its feasibility for quantitative analysis of methylation.

CONCLUSIONSYBR Green fluorescent PCR is an easy, fast and high-throughout quantitative tool, and it can be used for methylation analysis in basic research or clinical assay.

Actins ; genetics ; Biopsy ; Calibration ; Carcinoma, Hepatocellular ; genetics ; pathology ; DNA Methylation ; Feasibility Studies ; Female ; Fluorescence ; Genes, p16 ; Humans ; Luminescent Measurements ; Male ; Middle Aged ; Nucleic Acid Denaturation ; Polymerase Chain Reaction ; methods ; Sensitivity and Specificity ; Transition Temperature

Objective:At present, there are several modes of patent transformation of hospital service invention in our country. However, most of them are one-to-one style which is time-consuming and difficult to achieve industrialization in a short time. This study focuses on whether it is appropriate for public hospitals to use the mode of general license (i.e. one patent licensed to several enterprises concurrently or successively) to turn the patents of invention into productivity more efficiently.Methods:Combined with the period of COVID-19 pandemic, Zhongshan Hospital Fudan University successfully applied the mode of general license to achieve the patent transfer experience, and discussed the important value of the mode of general license for translational medicine.Results:The latest achievements of medical research are turned into clinical medical technologies and products quickly and effectively, proposing an appropriate scheme for the rapid transformation of patents, which balanced the interest of different stakeholders to gain the win-win situation.Conclusions:The innovative general licensing model can enable the patents transformation quickly and efficiently as long as it′s used properly.

Objective:Patent is one of the most common carriers of medical research innovation. The development of translational medicine is the general trend. It is necessary to explore patent transformation using multiple transactions in public hospitals. This study focused on whether it is appropriate for public hospitals to transfer scientific achievements by equity valuation.Methods:To study the transformation of scientific achievements of state-owned intangible assets in China, summarize the common transaction methods of scientific achievements in colleges, universities as well as scientific research institutions according to literature review and policy analysis, combined with the state-owned hospital’s asset management measures.Results:Through comparative analysis of the differences among public hospitals, universities and scientific research institutions, taking into consideration of the regulations on intangible assets management of hospitals, it is confirmed that public hospitals are not suitable for the transformation of scientific achievements by the way of equity pricing in current. However, other patent transfer and patent license methods are feasible, which can effectively promote the development of translational medicine alternatively.Conclusions:In view of the particularity of public hospitals, public hospitals need to explore multiple transactions to suit for the transformation of hospital patents. Although the current stage of equity pricing transfer does not conform to the asset management regulations of public hospitals, there are other ways to choose at this stage.