Metal-free visible-light-driven cascade cyclization reaction to synthesize 2-oxindoles benzoyl and phenylsulfinyl radicals with acrylamide derivatives.

A metal-free visible-light-driven cascade cyclization reaction to synthesize 3-methyl-3-acetophenone-2-oxindoles and 3-methyl-3-(methylsulfonyl)benzene-2-oxindoles in yields up to 96% and 99%, benzoyl and phenylsulfinyl radicals with acrylamide derivatives is reported, respectively. Extensive studies, including gram-scale, radical capture and isotope experiments, were performed to indicate that the reaction may involve a radical process.

[The development of FSCLZLY--an ascites-ultrafiltration instrument].

It's known to all that the refractory ascites treatment has so far been a very difficult clinical problem. We have extracted much experience from the practical techniques used in the refractory ascites treatments of more than 1,000 cases, and have developed the ascites ultrafiltration & concentration therapeutic instrument--FSCLZLY-A. The clinical applications show that it is very effective.

[The effect of envelope protein mutation on hepatitis B virus assemble].

Objective: To investigate the effect of envelope protein mutation on HBV assembly.

Methods: The envelope protein mutated vectors were constructed by the molecular clone in vitro, and then transfected transiently in the cell HepG2. The expression and secretion of S protein was assay by ELISA.

[Cloning of human telomerase reverse transcriptase promoter for targeted cancer therapy].

Objective: An human telomerase reverse transcriptase (hTERT) promoter was cloned to investigate the effect of simplex virus-thymidine kinase (HSV-tk) gene-ganciclovir (GCV) system under control of this promoter on lung cancer cells A549.

Methods: Specific primers were designed to amplify the core hTERT promoter from HepG2 genome. A set of expression vectors encoding LacZ or tk gene under control of hTERT or hCMV promoter was constructed through standard molecular cloning methods.

[Study of mouse marrow cells differentiation into a hepatocyte lineage in vitro].

Objective: To explore whether bone marrow stem cells (MSCs) from adult mice can be induced to differentiate into hepatocytes by hepatocyte growth factor (HGF) alone and the time phase characteristics in the differentiation progress.

Methods: Adult mouse MSCs were treated with or without 100 ng/ml HGF, on days 0, 7, 14, 21, and 28. The morphologic characteristics of the cells were examined; the albumin (ALB), AFP mRNA was analyzed sub-quantively using reverse transcription polymerase chain reaction (RT-PCR) and immumohistochemistry techniques.

[Replication and encapsidation of HBV mutants with the truncated C gene].

Objective: To evaluate the replication and encapsidation of HBV mutants with the truncated C gene.

Methods: The HBV mutants with the truncated C gene were constructed by molecular cloning and PCR-based deletion in vitro. The replication and encapsidation of HBV mutants were investigated by Southern blotting, PCR and real-time fluorescence PCR respectively after transfecting the HBV mutants plasmid into HepG2 cells by using liposome.

[Development of hygromycin-resistant packaging cell line for hepatitis B virus-derived vectors].

Objective: To cooperate with the study of HBV vector, hygromycin-resistant packaging cell line was developed that allows encapsidation of plasmids into HBV particles.

Methods: Free of packaging signal, HBV genome was inserted into plasmid pMEP4, which expresses the HBV structural proteins including core, pol and preS/S proteins. HepG2 cell lines were employed to transfect with the construct.

[Study on anti-HBV effects of genetically engineered replication-defective hepatitis B virus expressing dominant negative mutants of core protein].

Background: To explore the possibility of using HBV as a gene delivery vector, and to test the anti-HBV effects by intracellular expression of dominant negative mutants of core protein.

Methods: Two kinds of full length mutant HBV genome, which express Core-partial P and Core-S fusion protein, were transfected into HepG 2.2.

[The anti-HBV effect and mechanism of C gene truncated mutant in vitro].

Objective: To explore the effect and mechanism on HBV replication in C gene truncated mutant.

Methods: Protein expression of C gene truncated vector and wild C gene vector were assay by SDS-PAGE Western blot. Constructed C gene truncated expression vector was cotransfected with wild HBV genome; virus load was detected by PCR in the culture medium and the cell.

[Approach to transforming hepatitis B virus as a gene therapeutic vector].

Objective: To evaluate the possibility of hepatitis B virus (HBV) as a vector in liver-targeting gene therapy.

Methods: A fragment containing the small envelope gene of HBV was replaced with the reporter gene green fluorescent protein (GFP) to construct the recombinant HBV vector, which was transfected into HepG2 cells with liposome. The expression of GFP was observed with fluorescence microscope.

[Mitogenic effects of growth and differentiation factors on rat liver stem cell WB-F344 in vitro].

Objective: To explore the relationship between the proliferation, differentiation of rat hepatic stem like cell line WB-F344 and cytokines in vitro.

Methods: (3)H thymidine labelling of new synthesized DNA was used to examine the mitogenic responsiveness of WB-F344 cells to cytokines, western blot was used to study the expression of cytokines receptors on hepatic stem cells, and apoptotic cells were detected by Flow cytometry.

Results: WB-F344 cells showed a proliferative response to the cytokines of hepatocyte growth factor (HGF), epidermal growth factor (EGF), fibroblast growth factor (FGF), Insulin at the dose of 80 ng/ml, and the relative cpm values are 982.

[Anti-HBV effects of genetically engineered replication-defective HBV with combined expression of antisense RNA and dominant negative mutants of core protein and construction of first-generation packaging cell line for HBV vector].

Objective: To explore the possibility of using HBV as a gene delivery vector, and to test the anti-HBV effects by intracellular combined expression of antisense RNA and dominant negative mutants of core protein.

Methods: Full length of mutant HBV genome, which expresses core-partial P fusion protein and/or antisense RNA, was transfected into HepG2.2.