Search PubMed⌕ Search

Biomedical subjects

Fei Hao

Publications and source records attributed to Fei Hao.

11 recordsLinked to original sources
PubMed2025

Validation of caprine H11 and the Rosa26 platform for transgene integration via CRISPR-based system: investigations on stable transgene expression and genetic biosafety.

CRISPR/Cas9 technology is an efficient tool for site-specific livestock gene editing. However, to minimize potential disruption of host genome function, exogenous genes should be integrated into well-characterized genomic loci, such as H11 or Rosa26, which have been empirically validated for stable transgene expression. This study established a multi-dimensional assessment system to evaluate the biological applicability of the H11 locus and the widely used Rosa26 targeting platform as sites for targeted integration of exogenous genes in goats. Donor cells carrying the enhanced green fluorescent protein (EGFP) reporter gene at the H11 and Rosa26 loci were generated via CRISPR/Cas9-mediated homology-directed repair; this was followed by somatic cell nuclear transfer to produce transgenic cloned embryos and healthy offspring. Multi-dimensional analyses revealed the following. At the cellular level, there was stable and efficient EGFP expression at integration sites, with donor cells maintaining normal cell cycle progression, proliferation capacity, and apoptosis levels, and with no alterations in the transcriptional integrity of adjacent genes. At the embryonic level, there was sustained EGFP expression across pre-implantation embryonic stages, with developmental metrics statistically indistinguishable from wild-type embryos. Finally, at the individual level, cloned offspring exhibited growth phenotypes consistent with wild-type counterparts, and EGFP showed broad-spectrum expression in eight tissues. This study establishes the first CRISPR/Cas9-based crossscale (cellular-embryonic-individual) validation in goats, demonstrating that the H11 and Rosa26 loci support efficient and stable transgene integration in goats. These results provide a precise and predictable technical framework for livestock genetic improvement.

Animals↗
PubMed2006

[Clinical analysis of the infection with human papillomavirus in women].

BACKGROUND: To determine the relationship between human papillomavirus infection, cervical carcinoma, pre-cancerous lesion and condyloma acuminatum. METHODS: From January 2004 to August 2005, 1086 inpatients in department of dermatology and department of gynaecology and obstetrics in Southwest Hospital and No. 302 Hospital with cervical lesions and condyloma were reviewed. All specimens were detected for HPV-DNA using techniques of Gene Array and fluorogenic quantitative polymerase chain reaction (FQ-PCR). All detections of HPV-DNA were performed in the first admission before the patients underwent any examination or treatment. RESULTS: The positive rates of HPV-DNA detection were 100% in cervical intraepithelial neoplasia (CIN) I, and II and cervical carcinoma. Among these, the main subtype was HPV 16. But some of the patients were found to be positive for more than 2 subtypes of HPV. While the commonest HPV subtype was HPV 18 in endometrial cancer. Some of the patients were detected to be positive for more than 2 subtypes of HPV. In 636 female patient with condyloma acuminatum, the infection rates of HPV6, HPV11 accounted for 44.97% and 29.40%, respectively, HPV 16 and/or HPV 18 infection constituted a small percentage. In a few cases, infection with more than 2 subtypes was detected. CONCLUSION: Cervical carcinoma including pre-cancerous lesion differs from condyloma acuminatum in dominate infectious subtype of HPV. The former is mainly associated with HPV 16 and HPV 18 infections, and the latter mainly associated with HPV 6 and HPV 11 infections. But in both of the above lesions, a mixed infection with more than 2 types may occur and make the pathological changes and clinical treatment more complicated. The early diagnosis and supervision of HPV infection may be of great value for improvement of prognosis and quality of life.

Cervix Uteri↗
PubMed2005

Apoptosis of the dermal papilla cells of hair follicle associated with the expression of gene HSPCO16 in vitro.

The epithelial-mesenchymal interactions have an important role in the folliculomorphogenesis and mature hair follicle cycling. The dermal papilla, a dense aggregate of specialized dermis-derived stromal cells located at the bottom of the hair follicle, is a major component of hair, which signals the follicular epithelial cells to prolong the hair growth process. However, to date, little is known about the significance of the specific gene(s) expression in the dermal papilla cells with regard to their aggregative behaviour and hair cycling. In our previous study, the differentially gene-expressed cDNAs library had been determined by means of suppression subtractive hybridization technique between the aggregated human dermal papilla cells and control cells. Among those cDNAs library, the haematopoietic stem/progenitor cell (HSPC)-related gene HSPC016 was found. In this study, the gene HSPC016 was confirmed to express in the human dermal papilla cells by means of in situ hybridization and reverse transcriptase-polymerase chain reaction. In order to rudimentarily elucidate its biological function, a recombinant eucaryotic expressing plasmid pcDNA3.1(+)/HSPC016 was constructed and was transfected into the human dermal papilla cells and 3T3 fibroblast cells by means of Nucleofector(TM) technique (Amaxa, Cologne, Germany). Terminal deoxynucleotidyl transferase-mediated d-UTP nick end Labelling (TUNEL) assay showed that the expression of gene HSPCO16 resulted in the apoptosis of these cells, which suggested that the apoptosis of dermal papilla cells might be associated with the expression of gene HSPC016 in vitro.

3T3 Cells↗
PubMed2004

[Gene screening of dermal papilla cells in the state of aggregative growth and full-length cloning of HSPC016 gene for bioinformatics].

OBJECTIVE: To screen the genes of dermal papilla cells (DPC) related to the property of aggregative growth, and clone the full-length cDNA of differential HSPC016 gene for functional analysis. METHODS: DPC were collected from the hair of an individual aged 18 approximately 30 6 hours after the death. The complete papillae of hair were isolated and then cultured. The total DNA was extracted. Suppression subtractive hybridization-polymerase chain reaction was employed to screen out the genes differentially expressed in the DPC under the state of aggregative growth pattern in vitro. Then, rapid amplification of cDNA ends (RACE) technique was used to amplify the full-length cDNA of HSPC016 in the DPC, and bioinformatic methods were used to analyze their possible function. RESULTS: A subtractive library of human DPC was set up, and some up-regulated and down-regulated genes in the DPCs were screened out successfully. HSPC016 was identified to be a gene of 400 bp cDNA. Bioinformatic analyses and databases searching on the Internet indicated that this gene was mapped on chromosome 3 q21.31 and included an open reading frame with 195 bp coding for an expected 64aa soluble protein. The putative protein belonged to PD053992 family and was homologous to T2FA gene in domain. CONCLUSION: The establishment of subtractive library of DPC provides a solid foundation for further screening the genes related to aggregative growth and analyzing their regulatory mechanisms in DPC. Further more, HSPC016 in DPC may act as a subunit of a functional complex and play a role on transcriptional regulation within nucleus.

Adult↗
PubMed2004

Identification of differentially expressed genes in anagen dermal papilla by suppression subtractive hybridization.

BACKGROUND: We constructed a cDNA subtractive library of dermal papilla cells (DPCs) in anagen with suppression subtractive hybridization (SSH) technique and clone differentially expressed genes related to DPCs in anagen. METHODS: Total mRNA was isolated from DPCs of anagen and telogen follicles. Moreover, single-strand (ss) and double-strand (ds) cDNAs were synthesized in turn using SMART PCR cDNA synthesis technology. ds cDNAs then were digested with Rsa I and divided into two groups, and ligated to the specific adaptor 1 and adaptor 2R, respectively. After cDNAs were hybridized with each other twice and underwent two rounds of nested PCR. PCR products were ligated with arms of T/A plasmid vectors to set up the subtractive library. Selected clones were demonstrated by reverse Northern blot and sequenced. The acquired sequence data were aligned against the Genbank nucleotide database. RESULTS: cDNA subtractive library of DPCs in anagen follicles was set up successfully with high subtractive efficiency. Thirty-five genes were identified in this study with 22 known functional genes and 13 unknown functional genes. CONCLUSIONS: All results confirm the effectiveness and sensitivity of SSH in detecting differentially expressed genes from a small amount of clinical samples. Information about such alterations in gene expression could be useful for elucidating the genetic events in hair follicle growth regulation.

Adult↗
PubMed2004

[Retrospective analysis of 1905 patients with skin cancer from two general hospitals in western China from 1981 to 2000].

OBJECTIVE: To investigate the clinical features and changes in the incidence of skin cancer in two hospitals located in western China. METHODS: The patients diagnosed pathologically as skin cancer from 1981 to 2000 were retrospectively collected from the two hospitals. Clinical data of patients with skin cancer were collected and analyzed. RESULTS: (1) Of the 1 905 patients with skin cancer, squamous cell carcinoma accounted for 29.4%(560 patients), basal cell carcinoma 28.0% (534), and cutaneous malignant melanoma(CMM) 16.0% (305). (2) There were 591 patients with skin cancer diagnosed between 1980 and 1990, and 1 314 between 1991 and 2000, and accounted for 0.34% and 0.58% of all biopsy cases, respectively. The number of total biopsy patients increased 1.6% every year during the 20 years. The number of biopsy patients with skin cancer and with CMM increased 3.5% and 3.9% every year,respectively. (3) Of the 305 CMM patients, 63.3% located on the acra. These patients were elder, and have a higher rate of trauma and a higher incidence in the male than that in patients with CMM located on the other sites. (4) Of the 305 CMM patients, 64 (21%) had history of trauma at the primary onset sites, and 47 (15.4%) had history of small congenital nevi at the primary sites. CONCLUSION: There are some differences in the clinical features such as location and age between the skin cancer patients in our study and those in white population. The incidence of skin cancer in the two hospitals had been increasing in the 20 years (between 1981 and 2000). Both trauma and small congenital nevi are important risk factors of CMM.

Adult↗
PubMed2003

[In vitro infection of human liver cancer cell line HepG2 with HCV].

BACKGROUND: To test susceptibility of human liver cell line Hep G2 to HCV in vitro. METHODS: Hep G2 was cultivated with the serum from a chronic hepatitis C patient. After inoculation, plus and minus strand of HCV RNA, the expression of HCV NS3 antigen and the location of HCV RNA in cell and/or supernatant were examined by RT-PCR, immunohistochemistry and in situ hybridization, respectively. RESULTS: HCV RNA could be detected from day 2 to day 40 post-inoculation in both cell and supernatant. HCV NS3 antigen could be expressed in infected cells and HCV RNA was mainly situated within cytoplasm. CONCLUSIONS: The results suggested that HepG2 cell line was not only susceptible to HCV but also could support its replication in vitro.

Carcinoma, Hepatocellular↗
PubMed2002

[Study on point mutations in mitochondrial DNA control region for replication DLP(6) of cultured dermal fibroblast with 8-MOP/UVA treatment].

OBJECTIVE: To investigate the relationships between skin photoaging and point mutations in mitochondrial DNA (mtDNA) control region for replication of dermal fibroblast. METHODS: Cultured dermal fibroblasts were treated by 8-methoxypsora len /ultraviolet A (8-MOP/UVA). mtDNA was extracted by one-step-method and th e PCR products of D-loop and adjacent transcription promoter (DLP(6)) fragment of mtDNA control region for replication were detected by polymerase chain reaction-single strain conformation polymorphism and direct sequencing. RESULTS: After treated by 8-MOP/UVA, point mutations of 414 T-->G of DLP(6) fragment of mtDNA control region for replication largely accumulated. CONCLUSION: Accumulation of point mutations of DLP(6) fragment of mtDNA control region for replication may be closely associated with skin photoaging.

Adult↗
PubMed2002

[Effects of polypeptides from HCV core region on the function of cytotoxic T cells].

OBJECTIVE: To investigate the pathogenesis of cytotoxic T cell (CTL) dysfunction in patients with HCV infection. METHODS: BALB/c mice were immunized by subcutaneous injection of polypeptides from HCV core region, and the CTL activity of mouse spleen cells was detected by the LDH release test. Two polypeptides which can enhance CTL function and two polypeptides which can inhibit CTL function were selected and cross-combined. BALB/c mice were immunized using the combined polypeptides and the CTL activities were detected afterwards. RESULTS: CTL activity was inhibited by CPA9 (39-74 amino acids), CPB7 (67-76 amino acids) and CPB8 (71-80 amino acids), and promoted by CPA10 (5-23 amino acids), CPB6 (63-72 amino acids) and CPB2 (131-140 amino acids). Using single factor analysis of variance, the CTL activity in the mice could be enhanced by polypeptides from the HCV core region, CPB2+CPB8, CPB6+CPB8, respectively. There was no obvious difference between CPB2+CPB7, CPB6+CPB7 and negative control. CONCLUSIONS: CPA9, CPB7, and CPB8, the 3 polypeptides from HCV core region play an inhibition role and CPA10, CPB6, and CPB2 play an enhancement role in CTL activity in mice. The inhibition and enhancement functions of the polypeptides from HCV core region interact each other.

Animals↗
PubMed2000

Hepatitis C virus may infect extrahepatic tissues in patients with hepatitis C.

AIM:To explore the status of extrahepatic hepatitis C virus (HCV) infection and replication in hepatitis C patients,and its potential implication in HCV infection and pathogenicity.METHODS:By reverse-transcriptase poly-merase chain reaction (RT-PCR),in situ hybridization (ISH) and immunohis-tochemistry, HCV RNA, HCV replicative intermediate (minus-strand of HCV RNA), and HCV antigens were detected in 38 autopsy extrahepatic tissue specimens (including 9 kidneys, 9 hearts, 9 pancreas, 5 intestines, 2 adrenal glands, 2 spleens, 1 lymph node, and 1 gallbladder) from 9 hepatitis C patients, respectively; and the status of HCV replication in extrahepatic tissues was studied.RESULTS:By RT-PCR, all 9 patients were positive for HCV RNA in kidney, heart, pancreas, and intestine, but only 6(66.7%) patients were positive for HCV replicative intermediate. HCV RNA and HCV antigens were detected in kidney, heart, pancreas, intestine, adrenal gland, lymph node, and gallbladder in 5(55.6%) and 6(66.7%) patients by ISH and immuno-histochemistry, respectively. HCV RNA and HCV antigens were not detected in these extrahepatic organs in 3(33.3%) patients, although their livers were positive for HCV.HCV replicative intermediate detected by RT-PCR was consistent with HCV RNA and HCV antigens detected by ISH and immunohistochemistry (Kappa =0.42-0.75). HCV RNA and HCV antigens were detected in myocardial cells, epithelial cells of intestinal gladular, interstitial cells of kidney, epithelial cells of tubules and glomerulus, pancreas acinar cells and epithelial cells of pancreatic duct, epithelial cells of mucous membrane sinus of gallbladder, cortex and medulla cells in adrenal gland,and mononuclear cells in lymph node. HCV RNA was also detected in bile duct epithelial cells, sinusoidal cells, and mononuclear cells in liver tissues by ISH.CONCLUSION:HCV can infect extrahepatic tissues, and many various tissue cells may support HCV replication; extrahepatic HCV infection and replication may be of concomitant state in most of patients with hepatitis C. The infected extrahepatic tissues might act as a reservoir for HCV, and play a role in both HCV persistence and reactivation of infection. HCV as an etiologic agent replicating and expressing viral proteins in extrahepatic tissues itself contributes to extrahepatic syndrome associated-HCV infection in a few patients with chronic HCV infection.

Journal Article↗