Relationship between gene p53 codon 72 polymorphism and pathological scar formation after caesarean section / 中华整形外科杂志

<p><b>OBJECTIVE</b>To study the relationship between gene p53 codon 72 polymorphism and pathological scar formation occurrence after caesarean section.</p><p><b>METHODS</b>The method of molecular beacon with real-time PCR was applied to detect gene polymorphism of p53 codon 72 in blood samples taken from 303 pregnant women (within a week after caesarea section). The clinical visits were taken 3 times for 12th to 18th months to ascertain clinical formation of pathological scar and its relationship to genotype of p53. The chi-square method was used to analyze the relationship of p53 gene polymorphism and abnormal scar formation occurrence by statistical software SPSS 13.0.</p><p><b>RESULTS</b>Total of 303 pregnant women were assayed. 30 patients were found with pathological scar by clinical visit in the total 303 pregnant women. The genotype frequencies of total three types (C/C, C/G and G/G) of p53 gene codon 72 in patients with pathological scar are significantly different from that of normal pregnant woman. The frequency of C/C genotype in patients are higher than that of normal pregnant women (P < 0.01). The frequency of C/C genotype in these patients with pathological scar is higher (46.7%, 14/30) than C/G (33.0%, 10/30, P < 0.01) or G/G (20%, 6/30) genotype (P < 0.01). The C allele frequency in the patients is 63.7%. It is also higher than G allele (36.7%, P < 0.01). The OR value is 2.30. Therefore the C allele of p53 gene codon 72 is a risk factor for pathological scar.</p><p><b>CONCLUSIONS</b>There was a certain relationship between p53 gene codon 72 C allele and pathological scar formation after caesarean section.</p>

Association between cell apoptosis and the quality of early mouse embryos / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the relationship between cell apoptosis and the quality of early mouse embryos, understand the significance of apoptosis-regulatory genes in early embryonic development, and explore a new approach to improving the embryo quality.</p><p><b>METHODS</b>The levels of cell apoptosis and proliferation in early mouse embryos in different developmental status (morphologically normal embryos, arrested embryos and fragmented embryos) were analyzed with terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL), caspase in situ fluorescence and Bcl-2 immunofluorescence, and immunofluorescent detection of proliferating cell nuclear antigen (PCNA).</p><p><b>RESULTS</b>The cells in arrested embryos and embryonic fragments showed positive results in TUNEL assay with enhanced caspase activity and lowered expressions of Bcl-2 and PCNA.</p><p><b>CONCLUSION</b>Cell apoptosis in early mouse embryos may be closely related to embryonic arrest and fragmentation.</p>

Establishment of molecular beacon real-time PCR for detecting p53 gene single nucleotide polymorphism / 南方医科大学学报

<p><b>OBJECTIVE</b>To establish a method based on molecular beacon real-time PCR for detecting single nucleotide polymorphisms (SNP) in codon 72 of scar-related p53 gene.</p><p><b>METHODS</b>Two fluorescence-labeled molecular beacon probes were synthesized targeting CCC/CGC SNP of p53 codon 72. The genomic DNA was extracted from the peripheral blood of 28 patients with keloid, and the CCC/CGC SNP of P53 gene codon 72 were assayed with molecular beacon real-time PCR. The results of SNP typing were compared with the results of reverse dot hybridization and confirmed by direct DNA sequencing.</p><p><b>RESULTS</b>The goodness of fit of this method was 100% in comparison with direct DNA sequencing, higher than that of reverse dot hybridization.</p><p><b>CONCLUSION</b>Molecular beacon real-time PCR is suitable for rapid clinical detection of SNPs in p53 gene.</p>

Inhibitory effect of RNA interference targeting BaxBak on apoptosis of human granulosa cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the inhibitory effect of small interfering RNA (siRNA) targeting Bax-Bak on the apoptosis of human granulosa cells.</p><p><b>METHODS</b>Human granulosa cells were transfected with Bax-siRNA and Bak-siRNA either alone or in comibnation, and the cell morphological changes were obsered and the cell apoptosis was detected with flow cytometry. Western blotting was performed to examine the changes in Bax and Bak expressions in the transfected cells.</p><p><b>RESULTS</b>Western blotting demonstrated significantly weakened expressions of Bax and Bak in the transfected cells. The cell morphology of the cells tranfected with Bak siRNA and with both Bak and Bax siRNA remained normal; the cells with exclusive Bax siRNA transfection presented with basically normal cell morphology, but black spots were noted in the cytoplasm. In the positive and negative control groups, the cells became rounded and shrank with expanded intercellular spaces and numerous black spots in the cytoplasm. Flow cytometry showed apoptotic indexes of 3.44% and 3.97% in cells transfected with Bak siRNA and Bax-Bak siRNA, respectively, significantly lower than that in the negative group. Bax siRNA transfection resulted in an apoptotic index of 19.98%, similar to that in the negative group.</p><p><b>CONCLUSION</b>Interference of the expression of Bak gene inhibits the apoptosis of human granulosa cells, and the inhibitory effect can be enhanced by simultaneous Bax interference, which, when used alone, does not obviosuly inhibit the apoptosis of human granulosa cells.</p>

Expression of oxyntomodulin in bifidobacteria and effect of oxyntomodulin-transformed bifidobacteria on the body weight of obese mice / 南方医科大学学报

<p><b>OBJECTIVE</b>To observe the effect of pBBADs-OXM-transformed bifidobacteria on the body weight of obese mice.</p><p><b>METHODS</b>B. longum was transformed with pBBADs-OXM by electroporation, and arabopyranose-induced oxyntomodulin expression by the bacterium was detected by ELISA. pBBADs-OXM-transformed bifidobacteria was administered orally obese mice on a daily basis with pBBADs-GFP-transformed bifidobacteria as the negative control, and the body weight changes of the mice were observed.</p><p><b>RESULTS</b>OXM was detected by ELISA not only in the supernatant but also the precipitant of the transformed bacterial culture. The body weight of the obese mice fed with pBBADs-OXM-transformed bifidobacteria decreased significantly compared with that of the mice in the obese model group (P<0.05).</p><p><b>CONCLUSION</b>Administration of pBBADs-OXM-transformed B.longum can reduce the body weight of obese mice.</p>

BRCA1 regulates progesterone receptors A and B protein expressions in breast cancer cells in vitro / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the regulatory role of BRCA1 in the expression of progesterone receptors A and B (PRA and PRB) in breast cancer cells.</p><p><b>METHODS</b>Breast cancer MCF-7 cells were transfected with pFlag-CMV2-BRCA1 wt plasmid containing a full-length BRCA1 cDNA or with BRCA1-specific siRNA via lipofectamine 2000 to induce overexpression or suppressed expression of BRCA1, respectively. Twenty-four hours after the transfection, the cells were incubated in fresh culture medium containing 100 nmol/L progesterone for 24 h. The total RNA extract or whole cell lysate was prepared for detecting BRCA1, PRA and PRB expressions using RT-PCR and Western blotting.</p><p><b>RESULTS</b>The protein expressions of PRA and PRB were significantly decreased whereas their mRNA expressions remained unchanged in MCF-7 cells overexpressing BRCA1. In MCF-7 cells with BRCA1 knock-down, in contrast, the PRA and PRB protein expressions were markedly increased.</p><p><b>CONCLUSION</b>In breast cancer cells, exogenous and endogenous BRCA1 can both down-regulate the expressions of PRA and PRB at the protein level.</p>

Antitumour effects on human colorectal carcinomas cells by stable silencing of phospholipase C-gamma 1 with lentivirus-delivered siRNA / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>In most colorectal carcinomas, the level of phospholipase C (PLC)-gamma 1 expression is greatly elevated. Increased expression of PLC-gamma 1 may play an important role in colon carcinogenesis, but the mechanism is not well known. The aim of this study was to evaluate the role of PLC-gamma 1 in colon carcinogenesis by using recombinant lentivirus that stably suppressed the PLC-gamma 1 expression in human colorectal carcinoma LoVo cells.</p><p><b>METHODS</b>Recombinant lentivirus producing PLC-gamma 1 siRNA were prepared. After LoVo cells were transduced by each lentivirus, stably transduced cells were selected by Blasticidin. The protein and mRNA expression of PLC-gamma 1 were examined by Western-blot and reverse transcription-polymerase chain reaction (RT-PCR) analysis, and the effects of the lentivirus on the cell adhesion, migration and apoptosis were analyzed.</p><p><b>RESULTS</b>Stable LoVo cell line deficient in PLC-gamma 1, was established. Notably, PLC-gamma 1 was silenced without affecting the levels of other subtypes of PLC so that the role of PLC-gamma 1 in colon carcinogenesis could be examined. Silencing of endogenous PLC-gamma 1 resulted in efficient inhibition of the adhesion and migration of LoVo cells in vitro and a great increase of 5-fluorouracil induced apoptosis (30%-40%) of LoVo cells.</p><p><b>CONCLUSIONS</b>PLC-gamma 1 may play an important role in metastasis and anti-apoptosis in human colorectal carcinomas.</p>

The Procaryotic Expression, Purification and Activity Analysis of VIP-sTNFRII / 中国生物工程杂志

A prokaryotic expression plasmid containing VIP (vasoactive intestinal peptide) and sTNFRII(soluble tumor necrosis factor receptor II ) genes was constructed. The sTNFRII was cloned by PCR by using special primers which contained VIP gene ORF and a linker in its forward primer. The amplified fragment was inserted into the expression vector pET32a between BamHI and Hind III restriction sites. Transformed E.coli DH5 by pET32a-VIP- sTNFRIIexpressed the fusion protein. After being identified, the protein was purified by ion exchange chromatography and by hydrophobic interaction chromatography. The reconstructed protein showed high bio-activity and could be applied for further use.

p53 gene codon 72 polymorphism and susceptibility to keloid / 中华整形外科杂志

<p><b>OBJECTIVE</b>To investigate the relationship between p53 codon 72 polymorphism and susceptibility to keloid in a southern Chinese population.</p><p><b>METHODS</b>The p53 genotypes were determined by polymerase chain reaction-reverse dot blot (PCR-RDB) and DNA direct sequencing in 45 patients with keloid and 60 unrelated healthy controls.</p><p><b>RESULTS</b>The frequency of the p53 Pro allele among keloid patients was significantly higher than that among healthy controls (chi2 = 4.485, P = 0.034). The Pro/Arg and Arg/Arg genotype distribution among keloid patients was not significantly different from that among healthy controls (chi2 = 0.949, 1.346; P = 0.330, 0.246, respectively). However, the Pro/Pro genotype frequency among keloid patients was significantly higher than that among healthy controls (chi2 = 4.375, P = 0.036). The p53 Pro/Pro genotype significantly increased the risk for developing keloid, compared to the combination of Pro/Arg and Arg/Arg genotypes,with the odds ratio (OR) of 2.400 (95%CI: 1.048-5.498).</p><p><b>CONCLUSIONS</b>Determination of the p53 codon 72 genotype may be used as a stratification marker to predicate high-risk individuals for keloid.</p>