Biomedical subjects

Xiao-ke Hao

Publications and source records attributed to Xiao-ke Hao.

5 recordsLinked to original sources

PubMed2006

[Screening of human anti-gamma-seminoprotein light chain guided with the murine Fd fragment].

AIM: To screen human anti-gamma-sm (gamma-seminoprotein) light chain (Lc) with guided selection of murine Fd fragment. METHODS: The human Lc repertorie genes were amplified by RT-PCR from PBMC in patients with prostate cancer, and cloned into the phagemid vector pComb3X with murine Fd gene against gamma-seminoprotein to construct the human-mouse hybrid Fab antibody library. The size of the library, antibody gene recombinant percentage and diversity were identified by colony counting, plasmid digestion and colony sequence analysis, respectively. Purified gamma-sm was used as antigen to screen the displayed phage hybrid antibody library rescued by helper phage M13K07 for three rounds. The positive clones were selected by ELISA with pIII-fusion antibody and then the sequences of light gene in the positive clones were analyzed by IMGT-VQUEST. RESULTS: A 1.2x10(7) CPU human-mouse Fab antibody library was constructed with 90% Lc gene recombinant and great diversity. After 3 rounds' panning with gamma-sm, 5 positive clones were selected by ELISA and 2 clones with higher affinity were selected. Sequence analysis suggested these two positive clones contained the same light gene with high V(L) homology to human germline gene IGKV4-1*01. CONCLUSION: Human anti-gamma-sm light chain was successfully screened by constructing mouse-human hybrid Fab phage antibody library with murine Fd-guided selection.

Animals↗

PubMed2005

[Relationship between the amount of nitric oxide and apoptosis of germ cells in the semen of infertile men].

AIM: To explore the relationship between nitric oxide(NO) and apoptosis of germ cells in the semen of infertile men. METHODS: Copper-coated cadmium reduction fluorescence assay was used to detect nitrate, the metabolic product of NO. The apoptosis and ultrastructure of germ cells were detected and observed by TdT-mediated dUTP nick end labeling (TUNEL) and transmission electronls microscope (TEM), respectively. RESULTS: In normal control group, concentration of NO and the apoptotic rate of germ cells were (56.83+/-11.65) micromol/L and (4.60+/-1.25)%, respectively; while in inferile group were (128.86+/-23.76) micromol/L and (17.36+/-3.05)%, respectively. The differences in NO concentration and the apoptotic rate of germ cells between the two groups were significant (P<0.01). The concentration of NO was positively correlated with the apoptotic rate of germ cells in infertile group (r=0.96). The apoptotic germ cells showed typical morphology of apoptosis, including nuclear chromatin condensation and margination, nuclear memberane folding, and formation of apoptotic bodies. CONCLUSION: The amount of NO in semen from infertile men has close relationship with apoptotic rate of germ cells. High concentration of NO may result in the increase of germ cell apoptosis rate and lead to male infertility.

Adult↗

PubMed2005

[Study on the nucleic acid of E. coli bacteriophage with broad host range and its sterilization effect to sewage samples from the environment].

OBJECTIVE: To study the change of nucleic acid sequence and the germicidal effect of an E. coli bacteriophage with broad host range isolated from hospital sewage as well as to study the mechanism of phage host specificity and the effect of killed bacteria by phage-disinfectant to the samples from sewage water. METHODS: To extract the nucleic acid from phage f(2) and phage with broad host range using anti-serum-carbamidine hydrochloride assay. Purity with agarose gel electrophoresis was then evaluated. Differences of nucleic acid sequence between phage f(2) and phage with broad host range with reverse transcription-polymerase chain reaction (RT-PCR) and random amplified polymorphic DNA (RAPD)-PCR were also comparing and analysed. Through observing the germicidal test of phage f(2) and phage with broad host range to samples from environment, different sterilization effects between the two phages were compared. RESULTS: Analystic test for nucleic acid revealed that the two phages both belonged to 6000 bp, single-stranded RNA bacteriophage. Significant differences in their specificity of RAPD-PCR and RT-PCR were found during the changed of host range; with 26 RAPD-cDNA differential fragments found that in two phages RAPD-PCR products. The RT-PCR product of phage f(2) was 450 bp cDNA fragment, but the phage with broad host range did not show PCR product. Treating the sewage water with phage under broad host range, the germicidal test showed that the cleaning rate of E. coli bacteria and phage f(2) in water samples from environment could reach 36.75% - 56.28%, 30.84% - 47.96%, 19.19% - 35.06% and 13.05% - 27.85%, respectively. CONCLUSION: The cleaning rates to E. coli and bacteria by phage with broad host range were obviously higher than phage f(2) (P = 0.000). Analytic test for nucleic acid indicated that host-specific lytic effect of phage with broad host range had been changed at genetic level.

Bacteriophages↗

PubMed2004

[Study on isoniazid-resistant Mycobacterium tuberculosis isolates by multiple-polymerase chain reaction-single strand conformation polymorphism].

OBJECTIVE: To develop a new multiple-polymerase chain reaction-single strand conformation polymorphism (multi-PCR-SSCP) system for detecting the aphC promoter, inhA, and katG gene mutations in isoniazid-resistant Mycobacterium tuberculosis isolates in the single reaction, and for the quick diagnosis of isoniazid-resistant Mycobacterium tuberculosis isolates. METHODS: Three pairs of oligonucleotide primers were designed according to the aphC promoter, inhA, and katG genes of Mycobacterium tuberculosis to examine isoniazid-resistance by multi-PCR-SSCP. RESULTS: Isoniazid-sensitivity and resistance were analyzed with general PCR and multi-PCR at the same time, and H(37) Rv was used as a control. These two protocols amplified the anticipated fragments, the rate of consistency being 100%. By single gene PCR-SSCP, the mutation rates of aphC promoter, inhA, and katG gene were 17%, 20%, and 66%, respectively. The mutation rate detected by multi-PCR-SSCP was 83%. CONCLUSIONS: Multi-PCR-SSCP is a sensitive and specific method for rapid detection of aphC promoter, inhA, and katG gene mutations in isoniazid-resistant Mycobacterium tuberculosis isolates. Drug-resistant gene detection may be clinically useful in the therapy of tuberculosis.

Antitubercular Agents↗

PubMed2003

[Prokaryotic expression and bioactivity identification of Fab against human gamma-seminoprotein].

AIM: To construct the expression vector containing cDNA encoding Fab against human gamma-seminoprotein and express it in E. coli. METHODS: The genes encoding K chain and Fd against gamma-seminoprotein were acquired from pUC19-K and pBluescript KS( M13-)-Fd by restrictive enzyme digestion and then cloned into the expression vector pComb3 to construct recombinant expression vector pComb3-Fab. pComb3-Fab was transfected into and expressed in XLI-Blue. RESULTS: Fab against r-semino-protein was expressed in. XLI-Blue. Western blot analysis and immunocytochemical staining demonstrated that ex-pressed Fab could specifically bind to gamma-seminoprotein. CONCLUSION: Fab against gamma -seminoprotein has been expressed successfully with biological activity, which create favourable condition for further study on targeted therapy of prostate cancer.

Animals↗