ObjectiveTo investigate the role and mechanism of DNA repair regulation in the process of hepatocellular carcinoma （HCC） recurrence. MethodsHCC tissue samples were collected from the patients with recurrence within two years or the patients with a good prognosis after 5 years， and the Tandem Mass Tag-labeled quantification proteomic study was used to analyze the differentially expressed proteins enriched in the four pathways of DNA replication， mismatch repair， base excision repair， and nucleotide excision repair， and the regulatory pathways and targets that play a key role in the process of HCC recurrence were analyzed to predict the possible regulatory mechanisms. The independent samples t-test was used for comparison of continuous data between two groups； a one-way analysis of variance was used for comparison between multiple groups， and the least significant difference t-test was used for further comparison between two groups. ResultsFor the eukaryotic replication complex pathway， there were significant reductions in the protein expression levels of MCM2 （P=0.018）， MCM3 （P=0.047）， MCM4 （P=0.014）， MCM5 （P=0.008）， MCM6 （P=0.006）， MCM7 （P=0.007）， PCNA （P=0.019）， RFC4 （P=0.002）， RFC5 （P<0.001）， and LIG1 （P=0.042）； for the nucleotide excision repair pathway， there were significant reductions in the protein expression levels of PCNA （P=0.019）， RFC4 （P=0.002）， RFC5 （P<0.001）， and LIG1 （P=0.042）； for the base excision repair pathway， there were significant reductions in the protein expression levels of PCNA （P=0.019） and LIG1 （P=0.042） in the HCC recurrence group； for the mismatch repair pathway， there were significant reductions in the protein expression levels of MSH2 （P=0.026）， MSH6 （P=0.006）， RFC4 （P=0.002）， RFC5 （P<0.001）， PCNA （P=0.019）， and LIG1 （P=0.042） in recurrent HCC tissue. The differentially expressed proteins were involved in the important components of MCM complex， DNA polymerase complex， ligase LIG1， long patch base shear repair complex （long patch BER）， and DNA mismatch repair protein complex. The clinical sample validation analysis of important differentially expressed proteins regulated by DNA repair showed that except for MCM6 with a trend of reduction， the recurrence group also had significant reductions in the relative protein expression levels of MCM5 （P=0.008）， MCM7 （P=0.007）， RCF4 （P=0.002）， RCF5 （P<0.001）， and MSH6 （P=0.006）. ConclusionThere are significant reductions or deletions of multiple complex protein components in the process of DNA repair during HCC recurrence.

Objective: To investigate the distribution and expression of T3SS virulence genes in clinically isolated Pseudomonas aeruginosa (P. aeruginosa) strains and its correlation with drug resistance. Methods: A total of 68 P. aeruginosa isolates were collected from the First Affiliated Hospital of Wenzhou Medical University in 2015. The antimicrobial susceptibility was detected by the agar dilution method. The distribution of virulence genes such as exoU, exoS, exoT and exoY from different isolates was detected by PCR. The expression levels of transcriptional regulator genes (ptrA and exsA) and effector-related genes (exoT and exoS) in some isolates were determined by real-time fluorescence quantitative PCR, and the Pearson correlation analysis was used to analyze the results. Results: The detection rates of exoT and exoY in 68 P. aeruginosa isolates were higher, accounting for 79.4% and 75.0%, respectively. The detection rate of exoT in wound-sourced isolates was significantly higher than that in sputum (97.0% vs 61.8%, P＜0.01). In addition, the genotype of exoU - /exoS + was the most common, accounting for 51.5% (35/68). The resistance rates of sputum-sourced isolates to imipenem and meropenem were significantly higher than that of wound-sourced isolates (47.1% vs 8.8%, 47.1% vs 14.7%, P＜0.01). The resistance rates of isolates carrying exoU gene to carbapenems, aminoglycosides and fluoroquinolones were higher than those of isolates carrying exoS, exoT or exoY genes. Pearson correlation analysis showed that the expression level of ptrA gene was negatively correlated with those of exoT, exoS and exsA genes (P＜0.05). Conclusion The P. aeruginosa isolates from our hospital carrying T3SS virulence genes exoT and exoY are common, and the virulence genes are related to the drug resistance of P. aeruginosa. In addition, ptrA may be a potential negative regulatory gene for the expression of T3SS virulence genes.

Objective To explore the accurate positioning of Kawase triangle area via intradural anterior subtemporal transpetrosal approach. Methods On 14 dry skulls, the highest point of arcuate eminence (A), the junction of lateral margin of petrous ridge and anterior margin of transverse sinus (J), the petrous apex (P), the outermost point of sulcus nervi petrosi superficialis majoris (B), the outermost point of foramen spinosum (C), the outermost point of foramen ovale (D), the outermost point of trigeminal notch (E), and the outermost point of foramen lacerum (F) were marked. The distances of JA, JB, JC, JD, JE and JF were measured using point J as reference point. Using the line (JP) between point J and point P as the baseline, the angles of baseline with JA, JB, JC, JD, JE, and JF were measured. Results There were no significant differences in the distances of JA, JB, JC, JD, JE and JF between the left and right sides (P>0.05). There were no significant differences in the angles of the baseline with JA, JB, JC, JD, JE, and JF between left and right sides (P>0.05). Conclusion Using point J and the baseline JP as referent indexes, bony structures can be precisely located via intradural anterior subtemporal transpetrosal approach to orientate the Kawase triangle area; this method can insure rapid, accurate and safe drilling of anterior petrosal bone and exposing of petroclival region.

Objective To discuss the clinical efficacy of surgery for chronic subdural hematoma assisted by rigid neuroendoscope and its surgical techniques. Methods Clinical data of 161 patients with chronic subdural hematoma from August 2009 to December 2015 was analyzed retrospectively. 74 of them experienced surgeries assisted by rigid neuroendoscope (endoscope group) and other 87 cases were operated without neuroendoscope (routine group) during the same period. Results Although there were significant difference in operative duration between the two groups, complications, ratio of total removal of hematoma after surgery, postoperative inpatient duration and recurrent rate of hematoma were more advantageous in endoscope group. The operative duration of endoscope group with (112.68 ± 34.86) min was longer than that of routine group with (74.11 ± 28.23) min (t = 7.75, P = 0.000), while the postoperative inpatient duration of endoscope group with (8.23 ± 2.01) d was shorter than that of another group with (10.79 ± 5.02) d (t = -4.12, P = 0.000). There were no surgical associated complications in endoscope group, but 1 patient in routine group experienced intracerebral hematoma of frontal lobe and associated aphemia. Total removal of hematoma was confirmed in endoscope group with 98.65% (73/74), which was higher than that in routine group with 86.21% (75/78) (χ2 = 8.34, P = 0.004). Hematoma recurrence was found in 16 cases of routine group (18.39%), but more superiority in endoscope group with 1.35% (χ2 = 12.29, P = 0.000). Outpatient follow-up was carried out in all patients from 6 to 38 months with an average duration of 30.06 months. In 17 cases with recurrent hematoma during follow-up, 15 of them were cured by a second surgery, and another 2 patients were cured by atorvastatin. Conclusion As a simple, safe and effective technique, the application of rigid neuroendoscope during surgery for chronic subdural hematoma is more advantage than routine surgery. A self-made suction with adjustable soft curved tip is suitable for such procedure.

OBJECTIVEThe aim of this study was to clarify whether the fusion of bone marrow mesenchymal stem cells (MSCs) with tumor cells can promote tumor angiogensis.

METHODSHuman glioma stem/progenitor cells (GSPCs) (SU3 cells) were transfected with red fluorescent protein (RFP) gene. Bone marrow mesenchymal stem cells (MSCs) were harvested from nude mice with whole-body green fluorescent protein (GFP) gene expression. Then the two kinds of cells were co-cultured in vitro. At the same time SU3-RFP was transplanted into the brain of GFP-expressing nude mice to establish xenograft tumors. The co-cultured cells, GFP/RFP double positive (yellow) cells and blood vessels obtained from the xenograft tumors were observed under fluorescent microscope and laser scanning confocal microscope.

RESULTSAfter five passages in vitro, MSCs maintained the proliferative activity and highly expressed CD105. CD105 was also expressed in the femurs of GFP-expressing nude mice, tumor cells, blood vessels of SU3 xenograft tumors, and clinical malignant gliomas. When MSCs were co-cultured with SU3-RFP, the ratio of yellow cells co-expressing RFP and GFP was significantly increased after extended time and continuous passages. According to the flow cytometry, yellow cells co-expressing RFP and GFP were 83.7% of the cultured cells. In tissue slices of the xenograft tumors, bundles of yellow vessel-like structure and cross-sectioned yellow vascular wall structures including vascular wall stroma cells were observed with RFP and GFP expression, and were identified as de novo formed vessels derived from fusion of MSCs with SU3-RFP cells.

CONCLUSIONCell fusion occurs between tumor cells and host MSCs and it promotes tumor angiogenesis.

Animals ; Bone Marrow Cells ; physiology ; Cell Communication ; Cell Fusion ; Cells, Cultured ; Glioma ; Green Fluorescent Proteins ; Humans ; Luminescent Proteins ; Mesenchymal Stromal Cells ; Mice ; Mice, Nude ; Microscopy, Fluorescence ; Neoplasms ; Neovascularization, Pathologic ; Stem Cells ; Transfection ; Transplantation, Heterologous

OBJECTIVETo investigate the polymorphism in the promoter region of PCA3 gene and its relationship with risk of prostate cancer (PCa).

METHODSThe promoter region of PCA3 gene of the DNA of peripheral blood mononuclear cells was detected by sequence analysis in the 186 PCa and 141 BPH patients and 135 healthy control individuals. If the samples were detected with polymorphism of insection/deletion, clone sequence analysis was used with pBS-T carrier to verify it.

RESULTSThere were 5 polymorphisms. TAAA repeat times: 4, 5, 6, 7, 8, and 8 genotypes (TAAA 4/5, TAAA 4/6, TAAA 5/5, TAAA 5/6, TAAA 5/7, TAAA 5/8, TAAA 6/6, and TAAA 6/7) were detected in the promoter region of PCA3 gene. The eight genotypes were divided into three groups: ≤10TAAA, 11TAAA, ≥12TAAA. Unconditional logistic regression analysis models were used to analyze the relationship between different genotypes and cancer risks adjusted by sex and age. The type 11TAAA and ≥12TAAA was associated with higher relative risk for prostate cancer than the group ≤10TAAA [OR=1.74, 95% CI=1.06-2.87 (for type 11TAAA); OR=5.63, 95% CI=1.85-17.19 (for type ≥12TAAA)]. In the 186 PCa patients, there was 62.4% allele of PCA3 gene with AG/CA mutation found in the promoter 18-19 bp region of PCA3 gene and it had a close relation with the development of prostate cancer.

CONCLUSIONSShort tandem repeats are found in the promoter region of the PCA3 gene in PCa patients, and the increase of TAAA repeat sequences highly enhance the relative risk of prostate cancer development. The occurrence of such STR might be related to the mutations in their upstream loci.

Antigens, Neoplasm ; genetics ; metabolism ; Base Sequence ; Genes, Neoplasm ; physiology ; Genotype ; Humans ; Leukocytes, Mononuclear ; Male ; Microsatellite Repeats ; Mutation ; Polymorphism, Genetic ; Promoter Regions, Genetic ; Prostatic Neoplasms ; epidemiology ; genetics ; Risk

Objective To investigate the polymorphism in the promoter region of PCA3 gene and its relationship with risk of prostate cancer ( PCa) . Methods The promoter region of PCA3 gene of the DNA of peripheral blood mononuclear cells was detected by sequence analysis in the 186 PCa and 141 BPH patients and 135 healthy control individuals. If the samples were detected with polymorphism of insection/deletion, clone sequence analysis was used with pBS?T carrier to verify it. Results There were 5 polymorphisms. TAAA repeat times:4, 5, 6, 7, 8, and 8 genotypes (TAAA 4/5, TAAA 4/6, TAAA 5/5, TAAA 5/6, TAAA 5/7, TAAA 5/8, TAAA 6/6, and TAAA 6/7) were detected in the promoter region of PCA3 gene. The eight genotypes were divided into three groups: ≤10TAAA, 11TAAA, ≥12TAAA. Unconditional logistic regression analysis models were used to analyze the relationship between different genotypes and cancer risks adjusted by sex and age. The type 11TAAA and≥12TAAA was associated with higher relative risk for prostate cancer than the group ≤10TAAA [ OR=1. 74, 95%CI=1. 06?2. 87 ( for type 11TAAA);OR=5. 63, 95%CI=1. 85?17. 19 (for type≥12TAAA)]. In the 186 PCa patients, there was 62. 4% allele of PCA3 gene with AG/CA mutation found in the promoter 18?19 bp region of PCA3 gene and it had a close relation with the development of prostate cancer. Conclusions Short tandem repeats are found in the promoter region of the PCA3 gene in PCa patients, and the increase of TAAA repeat sequences highly enhance the relative risk of prostate cancer development. The occurrence of such STR might be related to the mutations in their upstream loci.

Objective The aim of this study was to clarify whether the fusion of bone marrow mesenchymal stem cells ( MSCs) with tumor cells can promote tumor angiogensis.Methods Human glioma stem/progenitor cells (GSPCs) (SU3 cells) were transfected with red fluorescent protein (RFP) gene.Bone marrow mesenchymal stem cells ( MSCs) were harvested from nude mice with whole-body green fluorescent protein (GFP) gene expression.Then the two kinds of cells were co-cultured in vitro.At the same time SU3-RFP was transplanted into the brain of GFP-expressing nude mice to establish xenograft tumors.The co-cultured cells, GFP/RFP double positive ( yellow ) cells and blood vessels obtained from the xenograft tumors were observed under fluorescent microscope and laser scanning confocal microscope.Results After five passages in vitro, MSCs maintained the proliferative activity and highly expressed CD105.CD105 was also expressed in the femurs of GFP-expressing nude mice, tumor cells, blood vessels of SU3 xenograft tumors, and clinical malignant gliomas.When MSCs were co -cultured with SU3-RFP, the ratio of yellow cells co-expressing RFP and GFP was significantly increased after extended time and continuous passages. According to the flow cytometry, yellow cells co-expressing RFP and GFP were 83.7%of the cultured cells. In tissue slices of the xenograft tumors, bundles of yellow vessel-like structure and cross-sectioned yellow vascular wall structures including vascular wall stroma cells were observed with RFP and GFP expression, and were identified as de novo formed vessels derived from fusion of MSCs with SU3-RFP cells.Conclu sion Cell fusion occurs between tumor cells and host MSCs and it promotes tumor angiogenesis.

Objective To investigate the polymorphism in the promoter region of PCA3 gene and its relationship with risk of prostate cancer ( PCa) . Methods The promoter region of PCA3 gene of the DNA of peripheral blood mononuclear cells was detected by sequence analysis in the 186 PCa and 141 BPH patients and 135 healthy control individuals. If the samples were detected with polymorphism of insection/deletion, clone sequence analysis was used with pBS?T carrier to verify it. Results There were 5 polymorphisms. TAAA repeat times:4, 5, 6, 7, 8, and 8 genotypes (TAAA 4/5, TAAA 4/6, TAAA 5/5, TAAA 5/6, TAAA 5/7, TAAA 5/8, TAAA 6/6, and TAAA 6/7) were detected in the promoter region of PCA3 gene. The eight genotypes were divided into three groups: ≤10TAAA, 11TAAA, ≥12TAAA. Unconditional logistic regression analysis models were used to analyze the relationship between different genotypes and cancer risks adjusted by sex and age. The type 11TAAA and≥12TAAA was associated with higher relative risk for prostate cancer than the group ≤10TAAA [ OR=1. 74, 95%CI=1. 06?2. 87 ( for type 11TAAA);OR=5. 63, 95%CI=1. 85?17. 19 (for type≥12TAAA)]. In the 186 PCa patients, there was 62. 4% allele of PCA3 gene with AG/CA mutation found in the promoter 18?19 bp region of PCA3 gene and it had a close relation with the development of prostate cancer. Conclusions Short tandem repeats are found in the promoter region of the PCA3 gene in PCa patients, and the increase of TAAA repeat sequences highly enhance the relative risk of prostate cancer development. The occurrence of such STR might be related to the mutations in their upstream loci.

Objective The aim of this study was to clarify whether the fusion of bone marrow mesenchymal stem cells ( MSCs) with tumor cells can promote tumor angiogensis.Methods Human glioma stem/progenitor cells (GSPCs) (SU3 cells) were transfected with red fluorescent protein (RFP) gene.Bone marrow mesenchymal stem cells ( MSCs) were harvested from nude mice with whole-body green fluorescent protein (GFP) gene expression.Then the two kinds of cells were co-cultured in vitro.At the same time SU3-RFP was transplanted into the brain of GFP-expressing nude mice to establish xenograft tumors.The co-cultured cells, GFP/RFP double positive ( yellow ) cells and blood vessels obtained from the xenograft tumors were observed under fluorescent microscope and laser scanning confocal microscope.Results After five passages in vitro, MSCs maintained the proliferative activity and highly expressed CD105.CD105 was also expressed in the femurs of GFP-expressing nude mice, tumor cells, blood vessels of SU3 xenograft tumors, and clinical malignant gliomas.When MSCs were co -cultured with SU3-RFP, the ratio of yellow cells co-expressing RFP and GFP was significantly increased after extended time and continuous passages. According to the flow cytometry, yellow cells co-expressing RFP and GFP were 83.7%of the cultured cells. In tissue slices of the xenograft tumors, bundles of yellow vessel-like structure and cross-sectioned yellow vascular wall structures including vascular wall stroma cells were observed with RFP and GFP expression, and were identified as de novo formed vessels derived from fusion of MSCs with SU3-RFP cells.Conclu sion Cell fusion occurs between tumor cells and host MSCs and it promotes tumor angiogenesis.