OBJECTIVETo study the relationship between gene p53 codon 72 polymorphism and pathological scar formation occurrence after caesarean section.

METHODSThe 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.

RESULTSTotal 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.

CONCLUSIONSThere was a certain relationship between p53 gene codon 72 C allele and pathological scar formation after caesarean section.

Alleles ; Cesarean Section ; Cicatrix ; genetics ; Codon ; Female ; Gene Frequency ; Genes, p53 ; Genotype ; Humans ; Polymorphism, Genetic ; Pregnancy ; Risk Factors

Objective：The aim of this study was to determine the expression of p21waf1, c erbB 2, and p53 in node negative breast cancer in relation to the clinicopathological parameters and prognosis. Methods： Expression of p21waf1, c erbB 2 and p53 in 121 patients were determined by LSAB immunohistochemical method, and Kaplan Meier method and Cox proportional hazard model were used to analyze the relationship of their expressions with prognosis. Results： (1) The expression rate of p21waf1 was 48.8％ , and was in connection with histological grade and ER positive. (2) The expression rate of p53 and c erbB 2 was 36.4％ and 26.4％ respectively. C erbB 2 positive expression was associated with histological grade. (3) the expression of p21waf1 was inversely correlated with that of p53 protein (P< 0.05). (4) the results of survival analysis with Kaplan Meier method showed that better disease free survival(DFS) in the patients with p21waf1 expression than those without, the negative c erbB 2 expression exhibited a significantly better prognosis than positive c erbB 2 expression (P< 0.01). Conclusions： There was a significant correlation between the expression of p21waf1, c erbB 2,and tumor grade; a p53 indenpent pathway can mainly induce the expression of p21waf1; p21waf1 expression was a good prognostic marker for the patients with node negative breast cancer; c erbB 2 positive expression was an independent prognostic marker.

BACKGROUNDMutation and expression change of p21WAF1/CIP1 may play a role in the growth of osteosarcoma. This study was to investigate the expression of the p21WAF1/CIP1 gene in human osteosarcoma, p21WAF1/CIP1 gene DNA sequence change and their relationships with the phenotype and clinical prognosis.

METHODSp21WAF1/CIP1 gene in 10 normal people and the tumours of 45 osteosarcoma patients were examined using polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) with silver staining. The PCR product with an abnormal strand was sequenced directly. The p21WAF1/CIP1 gene mRNA and P21 protein of 45 cases of osteosarcoma were investigated by using in situ hybridization and immunohistochemistry, respectively.

RESULTSThe occurrence of P21 protein in osteosarcoma was 17.78% (8/45), and p21WAF1/CIP1 mRNA expression in osteosarcoma was 42.22% (19/45). The p21WAF1/CIP1 gene DNA sequencing of amplified production showed that in p21WAF1/CIP1 gene exon 3 of 36 cases of human osteosarcoma, there were 17 cases (47.22%) with C-->T at position 609; 10 normal blood samples' DNA sequence analysis yielded 8 cases (80.00%) with C-->T at the same position.

CONCLUSIONSAlong with the increase of malignancy, the expression of p21WAF1/CIP1mRNA and P21 protein in osteosarcoma tends to decrease. It is uncommon for the p21WAF1/CIP1 gene mutation to occur in human osteosarcoma. As a result, the possible existence of tumour subtypes of p21WAF1/CIP1 gene mutation should be investigated. Our research leads to the location of p21WAF1/CIP1 gene polymorphism of Chinese osteosarcoma patients, which can provide a basis for further research.

Cyclin-Dependent Kinase Inhibitor p21 ; Cyclins ; genetics ; Humans ; Osteosarcoma ; genetics ; Polymerase Chain Reaction ; Polymorphism, Single-Stranded Conformational ; RNA, Messenger ; analysis ; Sequence Analysis, DNA

OBJECTIVETo isolate Nanobacteria from dental pulp stone and perform culturing and the identification of Nanobacteria.

METHODSFreshly collected 27 dental pulp stones were divided into nine samples. Each sample contained three dental pulp stones. All samples were used for the isolation and culture of Nanobacteria. The shape and the growth characteristics of the cultured bacteria were observed. Nanobacteria were identified by von Kossa staining, immunohistochemical staining and indirect immunofluorescence staining, double staining including Hoechst staining and von Kossa staining.

RESULTSThe characteristics growth and morphology of the bacteria detected in seven samples were similar to Nanobacteria. von Kossa staining, immunohistochemical staining, indirect immunofluorescent staining were positive for Nanobacteria. In double staining method, Hoechst staining of the samples was negative for Nanobacteria, but von Kossa staining was positive. Hoechst staining of the dental pulp cells was positive. No Nanobacteria was found in the other two samples.

CONCLUSIONSThe bacteria isolated from dental pulp stone in this study was similar to Nanobacteria in terms of growth rate, morphology and staining properties. These unusual properties of the bacteria may play an important role in the formation of pulp stone.

Bacteria ; isolation & purification ; Dental Pulp Calcification ; microbiology ; Humans ; Immunohistochemistry ; In Vitro Techniques

OBJECTIVETo detect the inhibitory effect of all-trans retinoic acid(ATRA) on breast cancer stem cells (CSCs).

METHODSThe inhibitory effect of ATRA on MCF-7 and SK-BR-3 cell lines was analyzed using a Cell Counting Kit-8 (CCK-8). The proportion of CD44(+)CD24(-) tumor cells of the two cell lines were measured before and after the ATRA treatment, and the role of ATRA in the regulation of CSC self-renewing ability was evaluated with a tumor sphere assay. The tumor spheres were grown in an adherent culture to evaluate the ATRA-induced differentiation of breast cancer stem cells.

RESULTSATRA effectively inhibited the unsorted cells and stem cells, but the CSCs were more sensitive to ATRA. At a concentration of 10(-6) mol/L, the inhibitory rate of MCF-7 unsorted cells and stem cells were (8.66 ± 1.06)% and (21.09 ± 3.25)%, respectively (P = 0.004). For SK-BR-3 cells, the rates were (39.19 ± 1.47)% and (51.22 ± 2.80)%, respectively (P = 0.005). The self-renewing ability of the CSCs was impaired by ATRA at a concentration of 10(-6) mol/L. The rate of MCF-7 and SK-BR-3 stem cells to form tumor sphere was 5.2% (5/96) and 13.5% (13/96), respectively. For the control group, it was 86.5% (83/96) and 93.8% (90/96), respectively (P < 0.001). ATRA also promoted the CD44(+)CD24(-) subpopulation to differentiate. SK-BR-3 stem cells were grown in an adherent culture. After using ATRA, the proportion of CD44(+)CD24(-) cells was (48.1 ± 2.5)% and that of the control group was (86.6 ± 2.5)% (P < 0.001).

CONCLUSIONSATRA effectively inhibits breast NCSCs and CSCs, but CSCs are more sensitive to ATRA. ATRA impairs the self-renewing ability of CSCs and promotes CSCs to differentiate.

Antineoplastic Agents ; pharmacology ; Breast Neoplasms ; metabolism ; pathology ; CD24 Antigen ; metabolism ; Cell Differentiation ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Female ; Humans ; Hyaluronan Receptors ; metabolism ; Neoplastic Stem Cells ; cytology ; drug effects ; Tretinoin ; pharmacology

OBJECTIVETo establish a method based on molecular beacon real-time PCR for detecting single nucleotide polymorphisms (SNP) in codon 72 of scar-related p53 gene.

METHODSTwo 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.

RESULTSThe goodness of fit of this method was 100% in comparison with direct DNA sequencing, higher than that of reverse dot hybridization.

CONCLUSIONMolecular beacon real-time PCR is suitable for rapid clinical detection of SNPs in p53 gene.

Base Sequence ; Codon ; genetics ; Humans ; Keloid ; genetics ; Molecular Sequence Data ; Polymorphism, Single Nucleotide ; Real-Time Polymerase Chain Reaction ; methods ; Tumor Suppressor Protein p53 ; genetics

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.

OBJECTIVETo investigate the apoptosis and proliferation activity of Tca8113 cells.

METHODSThe vector that involves short hairpin RNA of iNOS was transfected to Tca8113 cells. The change of iNOS expression was observed using immunohistochemistry technique, the apoptosis rate examined by flow cytometry, and the proliferation Tca8113 cells examined by methyl thiazolyl tetrazolium (MTT).

RESULTSThe expression of iNOS in Psilencer-iNOS group was lower than that in control groups (P < 0.01), the apoptosis rate was higher than that in control groups (P < 0.01); whereas the proliferation activity of Tca8113 cells in Psilencer-iNOS group was lower than that in control groups.

CONCLUSIONSDown expression of iNOS by RNAi can promotes apoptosis of Tca8113 cells and has an anti-proliferation activity effect.

Apoptosis ; Carcinoma, Squamous Cell ; genetics ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Gene Expression Regulation, Neoplastic ; Genetic Vectors ; Humans ; Nitric Oxide Synthase Type II ; genetics ; metabolism ; Plasmids ; genetics ; RNA Interference ; Tongue Neoplasms ; genetics ; pathology ; Transfection

OBJECTIVEIdiopathic collapsing glomerulopathy (ICG) is a clinically and pathologically distinct variant of focal segmental glomerulosclerosis, which is characterized by proteinuria (often nephrotic range) and rapid progression to end-stage renal failure. The typical pathological changes are global/segmental glomerular collapse, hypertrophy and hyperplasia of podocytes and severe tubulointerstitial lesions. Most ICG patients who have been reported in previous published papers are adults. ICG in children is rare. The study aimed to analyze and investigate clinical manifestations, renal histopathological findings, treatment and outcomes of ICG in children.

METHODSData of two cases of ICG, a 7-year-old boy and a 12-year-old girl, were analyzed. Both of them were Chinese and Han. Clinical characteristics, results of laboratory tests, renal histopathological findings, treatment, outcomes and prognosis of the two children with ICG were retrospectively analyzed. Results were compared with published data.

RESULTSThese two children presented typical clinical features of nephrotic syndrome. The quantity of 24 hr urine protein was 7.6 g/d (0.47 g/kg x d for boy) and 10.67 g/d (0.35 g/kg x d for girl). Both of them had hypertension (blood pressure ranged from 130/90 to 150/110 mmHg) and hypercholesterolemia (15.4 mmol/L for the boy and 11.3 mmol/L for the girl). The serum albumin was 12 g/L for girl and 23 g/L for boy. The creatinine clearance rate gradually decreased from normal range to 30 ml/min for the girl. The histopathological changes in renal biopsy of them were focal segmental or global glomerular collapse, hypertrophy and hyperplasia of podocytes and severe tubulointerstitial lesions. These two cases were steroid-resistant and were treated with pulse intravenous methylprednisolone and pulse intravenous cyclphosphamade in one case, who rapidly progressed to end-stage renal failure and died half a year later. Another one was treated with cyclosporine. He showed continuous hypertention and heavy proteinuria for eight months.

CONCLUSIONICG in the 2 children was a severe disease which presented steroid-resistant nephrotic syndrome and rapidly progressive renal failure. The pathological characteristics was global/segmental glomerular collapse, hypertrophy and hyperplasia of podocytes and severe tubulointerstitial lesions. In children with ICG treatment was difficult and the prognosis was poor.

Child ; Disease Progression ; Female ; Glomerulosclerosis, Focal Segmental ; complications ; diagnosis ; pathology ; therapy ; Glucosinolates ; Humans ; Kidney ; pathology ; Kidney Failure, Chronic ; etiology ; Kidney Glomerulus ; pathology ; Male ; Nephrotic Syndrome ; etiology ; Proteinuria ; etiology ; Treatment Outcome

BACKGROUNDBrain metastasis has become one of the most important factors of the failure of treatment of locally advanced non-small cell lung cancer (LANSCLC). There is no conclusion whether NSCLC patients should receive prophylactic cranial irradiation (PCI) or not. The aim of this study is to analyze the risk factors of brain metastasis of LANSCLC after surgery to find out the sign of PCI for LANSCLC.

METHODSA total of 223 patients with stage III NSCLC who received surgical resection were retrospectively analyzed. The risk factors of brain metastasis were determined to set up a mathematic model for brain metastasis.

RESULTSThe median survival time after surgery was 28.0 months. The 1-, 2- and 3-year survival rate was 84.3%, 56.9% and 44.8% respectively. The incidence of brain metastasis was 38.1% (85/223). Patients with extensive mediastinal lymph node metastasis, more node metastasis and non-squamous carcinoma showed significantly higher incidence of brain metastasis than those with limited mediastinal lymph node metastasis, fewer positive mediastinal lymph nodes and squamous carcinoma (P=0.000, P=0.000, P=0.013). The mathematic model of brain metastasis was: logit(P)=8.215-0.903×NPN-0.872×RT-0.714×HG-1.893×LE-0.948×HS-1.034×PC (NPN=No. of positive nodes, RT=resection type, HG=histology, LE=location and extent of mediastinal lymph node metastasis, HS=histologic stage, PC=postoperative chemotherapy). P≥0.44 meant high risk for brain metastasis.

CONCLUSIONSHigh risk factors of brain metastasis in LANSCLC patients after complete resection of the cancer include non-squamous carcinoma, extensive and more mediastinal lymph node metastasis. P≥0.44 may be considered a sign of PCI in clinical trial.