PiggyBac transposon-mediated gene delivery efficiently generates stable transfectants derived from cultured primary human deciduous tooth dental pulp cells(HDDPCs) and HDDPC-derived iPS cells

The ability of human deciduous tooth dental pulp cells（HDDPCs） to differentiate into odontoblasts that generate mineralized tissue holds immense potential for therapeutic use in the field of tooth regenerative medicine. Realization of this potential depends on efficient and optimized protocols for the genetic manipulation of HDDPCs. In this study, we demonstrate the use of a Piggy Bac（PB）-based gene transfer system as a method for introducing nonviral transposon DNA into HDDPCs and HDDPC-derived inducible pluripotent stem cells. The transfection efficiency of the PB-based system was significantly greater than previously reported for electroporation-based transfection of plasmid DNA. Using the neomycin resistance gene as a selection marker, HDDPCs were stably transfected at a rate nearly 40-fold higher than that achieved using conventional methods. Using this system, it was also possible to introduce two constructs simultaneously into a single cell. The resulting stable transfectants, expressing td Tomato and enhanced green fluorescent protein, exhibited both red and green fluorescence. The established cell line did not lose the acquired phenotype over three months of culture. Based on our results, we concluded that PB is superior to currently available methods for introducing plasmid DNA into HDDPCs. There may be significant challenges in the direct clinical application of this method for human dental tissue engineering due to safety risks and ethical concerns. However, the high level of transfection achieved with PB may have significant advantages in basic scientific research for dental tissue engineering applications, such as functional studies of genes and proteins. Furthermore, it is a useful tool for the isolation of genetically engineered HDDPC-derived stem cells for studies in tooth regenerative medicine.

Inhibition effect of Chinese herbal medicine on transcription of hepatitis C virus structural gene in vitro

AIM: To investigate the inhibitory effect of Chinese herbal medicine on the transcription of hepatitis C virus (HCV) structural gene in Hela D cells.METHODS: Hela cell line was transfected with recombinant pBK-CMV-HCV containing HCV structural gene by Lipofectamine. RT-nested-PCR and Western blot assay were used to testify the HCV gene expression in Hela cells. The Hela cells expressing HCV structural protein were named Hela D cells. Prescriptions of Xiao chaihu Decoction (XCHD),Fufang Huangqi (FFHQ) and Bingganling (BGL) wererespectively added to Hela D cells in various concentrations. Semi-quantitative RT-nested-PCR product analysis was performed according to the fluorescent density between HCV DNA band and GAPDH DNA band in gel electrophoresisafter screened. RESULTS: Recombinant pBK-CMV-HCV could correctly express the HCV structural gene in Hela D cells. After coculture of Hela D cells with three prescriptional different concentrations for 48 h respectively, the transcription of HCVgene decreased with increasing of the concentration of each prescription. The lightness ratio of HCV product bands to GAPDH product bands was 0.24, 0.10 and 0.12 in Hela D cells incubated with 0.1 g/mL of XCHD, FFHQand BGL respectively and the lightness ratio HCV product bands to GAPDH product bands was 0.75, 0.67 and 0.61respectively in the control cells. CONCLUSION: The prescriptions of XCHD, FFHQ and BGL partly inhibit the transcription of HCV structural gene inHela D cells.

The clinical analysis of secondary hypertension patients

Objective To analyze the situation of secondary hypertension caused by different etiology and its treatment status so as to improve its treatment.Methods Totally 580 patients with secondary hypertension selected with strict criteria were divided into 4 groups in accordance with the etiology.The data were dealt with SPSS 13.0.We analyzed the sex,age,systolic pressure,diastolic pressure and positive incidence of family history of all the secondary hypertension patients in each group,and compared their usage of every unilateral preparation.Results The average age of renal parenchymal disease was more advanced than that of adrenal disease and other diseases.The administration of diuretics and angiotensin-converting enzyme inhibitor(ACEI)in renal parenchymal disease group,αβ receptor blocking agents and calcium antagonists in kidney-artery disease group,α antagonists in adrenal disease and compound antihypertensive agents in other diseases groups all affected advantage of the ratio.Conclusion Diuretics and ACEI are suitable for hypertension caused by renal parenchymal disease.αβ receptor blocking agents and calcium antagonists are suitable for that caused by kidney artery disease,but α receptor blocking agents are suitable for that caused by adrenal disease.

Development of a Kit Choosing Medicine Rapidly for Maricultural Bacteriosis

[ Objective] This study aimed to develop a kit to rapidly choose an appropriate medicine for maricultural bactedosis. [ Method] By screening enriched bacteria, solid medium and medicine and comparing effects of rapidly choosing medicine, a kit that could pick up the right medicine quickly for mariculture bacteriosis was created. [Result] The pilot test in some regions showed the kit had ideal yesalts and was easy to operate. [ Conclusion] The kit, a convenient and useful kit to detect drug sensitivity, can help users select effective drugs from many common medicines rapidly and accurately, and it is mainly suitable to use in the rapid selection of an appropriate medicine for maricultural bacteriosis.

Cancer stem cells, stemness markers and selected drug targeting: metastatic colorectal cancer and cyclooxygenase-2/prostaglandin E2 connection to WNT as a model system

Few studies have reported on the analyses of drugs targeting enriched populations of cancer stem cells (CSCs) as a means for identifying potent anti-CSC agents. This review evaluates recent information on the identification and functions of specific CSC surface markers, with particular emphasis on colorectal cancers and the screening of drugs to eliminate such cells. Many of these CSC markers are found commonly expressed on CSCs from different cancer types as well as embryonic stem cells. These markers are often related to hypoxic activation of the WNT/b-catenin pathway, cyclooxygenase-2/prostaglandin E signalling and their relationship to LGR5. By effectively using drugs that inhibit these pathways to kill the CSC population, or otherwise forcing them out of dormancy into active cell division, cancers should become more susceptible to chemotherapy. Such combinational therapies targeting both CSCs and proliferating tumor cells should greatly improve upon the current basis for treatment.

Study on the Drug Selection Law for Treatment of Chronic Gastritis with Spleen Deficiency and Stomach Dryness by Complex System Entropy Cluster

Objective:To study on Prof. GAO Zhong-ying’s drug selection law for treatment of chronic gastritis with data mining technique. Methods: The 407 medical records of chronic gastritis treated by Prof. GAO Zhong-ying were collected and the study on these drugs in the recipes was carried out with data mining method. Among them, the recipe composed of one drug was studied with frequency statistical method, correlativity between drug pairs with improved mutual information, correlativity among multi-drugs with complex system entropy cluster technique. Results: In treatment of chronic gastritis by Prof. GAO Zhong-ying there were 30 drugs with a higher use frequency of over 38 times, 94 commonly-used drug pairs with correlation coefficient of over 0.05, 11 commonly-used drug core combinations. Conclusion: The results attained with data mining technique for studying experience of famous and old TCM physicians conform to the clinical practice and the method is of an important significance for summarization of famous and old TCM physicians’ experiences.