Construction and detection of the tissue-specific pINV-HPV16 E6/7 vector.

A tissue-specific promoter can control downstream gene expression in tissues or organs. The human involucrin (hINV) promoter (pINV) that contains 2474 bp of hINV upstream sequence is able to regulate tissue-specific gene expression. This tissue specificity may be important for the prevention and treatment of human papilloma virus infections.

Identification of a novel mouse brachyury (T) allele causing a short tail mutation in mice.

Mutations in T-box genes are associated with numerous disease states in humans. The objective of this paper was to characterize the T(shao), a specific T-box mutation, in mice. T(shao), a short-tailed mutant mouse strain in a B6 background, was obtained by ethylnitrosourea mutagenesis.

[Construction and identification of recombinant adeno-associated virus expressing siRNA].

Adeno-associated virus (AAV) mediated RNA interference can be used to inhibit the expression of homologous genes in different mammalian cells. In this study, a transfer plasmid (pAAV-EGFP-H1) containing the H1 promoter and EGFP-expressing cassette was constructed based on the backbone of pAAV-MCS. Using calcium phosphate precipitation method, pAAV-EGFP-H1 was co-transfected into AAV-293 cells with helper plasmids and infective recombinant AAV was generated.

[Construction of Fab antibody against Rh antigen].

Aim: To Construct Fab antibody against Rh antigen.

Methods: The variable regions of light and heavy chains were amplified by RT-PCR from the PBMCs of volunteers with high titer (1:256-512 by inditect agglutation) antibody to Rh antigen. Meanwhile, the genes of constant regions of light and heavy chains were isolated from pComb3xTT and pComb3xlambda phagmid carrying the templates respectively.

[The effect of HPV16E7 DNA vaccine transdermal delivery with microneedle array].

Objective: To study the effects of DNA vaccine transdermal delivery with microneedle array.

Methods: The pcDNA3.1-HPV16E7 recombinant vector acting as gene vaccine was established.

Involvement of insulin-like growth factor-insulin receptor signal pathway in the transgenic mouse model of medulloblastoma.

A transgenic mouse model expressing Simian virus 40 T-antigen (SV40Tag) under the control of a tetracycline system was generated. In this model, a cerebellar tumor was developed after doxycycline hydrochloride treatment. Real time-polymerase chain reaction and immunohistochemistry results indicated that the SV40Tag gene was expressed in the tumor.

Generation and characterization of islet cell tumor in pTet-on/pTRE-SV40Tag double-transgenic mice model.

A line of double-transgenic mice that develop neoplasms arising primarily in the pancreas was established. In these mice, the oncogene SV40 T antigen (Tag) was detected in the pancreas with and without the control of Tet-on system. The transgenic mice that developed pancreatic tumors as early as 20 weeks of age showed hypoglycemia on a blood glucose test.

[Preparation and identification of monoclonal antibodies against Type M/N and Glycophorin A/B].

Aim: To prepare monoclonal antibodies(mAbs) against Type M/N and monoclonal antibodies against Glycophorin A and Glycophorin B (GPA/GPB) and identify rare blood group MkMk.

Methods: 8 week-old female BALB/c mice were immunized with type "O" red blood cells, Splenocytes from the immunized mice were fused with Sp2/0 myeloma cells, and positive hybridoma clones were screened by panel erythrocytes. The titer of supernatant and ascitic fluid was tested by direct and indirect agglutination.

Gene expression analysis of pancreatic cystic neoplasm in SV40Tag transgenic mice model.

Aim: To study the gene expression changes in pancreatic cystic neoplasm in SV40Tag transgenic mice model and to provide information about the prevention, clinical diagnosis and therapy of pancreatic cancer.

Methods: Using the pBC-SV40Tag transgenic mice model of pancreatic cystic neoplasm, we studied the gene expression changes by applying high-density microarrays. Validation of part gene expression profiling data was performed using real-time PCR.

Generation and characterization of a transgenic mouse model for pancreatic cancer.

Aim: To generate a SV40Tag transgenic tumor animal model and to study the mechanism underlying tumorigenesis.

Methods: A mammary gland expression vector containing SV40Tag DNA was generated. Transgene fragments were microinjeted into fertilized eggs of FVB mice.

Two kinds of ENU-induced scant hair mice and mapping of the mutant genes.

Objective: To establish mouse models for human diseases through N-ethyl-N-nitrosourea (ENU) mutagenesis, and to provide groundwork to clone genes and study their functions after mapping the mutant genes.

Methods: 18 male D2 mice (G0) at age of 8-10 weeks old were injected intraperitoneally with ENU (100 mg/kg) once a week for three consecutive weeks. The treated male mice were mated with females of the same strain, and their offspring (G1) were used to screen for dominant and recessive mutation.

Preliminary study on the production of transgenic mice harboring hepatitis B virus X gene.

AIM:To establish transgenic mice lineage harboring hepatitis B virus X gene and to provide an efficient animal model for studying the exact role of the HBx gene in the process of hepatocarcinogenesis.METHODS:The HBx transgenic mice were produced by microinjecting the construct with X gene of HBV (subtype adr) DNA fragment into fertilzed eggs derived from inbred C57BL/6 strain; transgenic mice were identified by using Nested PCR; expression and phenotype of HBx gene were analyzed in liver from transgenic mic at the age of 8 weeks by RT-PCR, pathologic examination and periodic acid-schiff staining (PAS), respectively.RESULTS:Five hundred and fourteen fertilized eggs of C57 BL/6 mice were microinjected with recombinant retroviral DNA fragment, and 368 survival eggs injected were transferred to the oviducts of 18 pseudopregnant recipient mice, 8 of them became pregnant and gave birth to 20 F1 offspring.