OBJECTIVETo investigate the feasibility of chondrogenesis in vitro with bone marrow stromal cells (BMSCs) induced by the co-cultured chondrocytes.

METHODSThe BMSCs and chondrocytes were separated from pig and cultured. The supernatant of chondrocytes was used as the inducing solution for BMSCs from the 2nd generation. 7 days later, samples were taken and underwent immunohistochemistry and RT-PCR for detection of the expression of specific type II cartilage collagen, type II collagen and aggrecan mRNA. The cultured BMSCs and chondrocytes were mixed at a ratio of 8:2 (BMSC: cartilage cell) and were inoculated into a polyglycolic acid/polylactic acid (PGA/PLA) scaffold at the final concentration of 5.0 x 10(7)/ml. The cartilage cells and BMSCs were also inoculated separately at the same concentration as the positive and negative control. Pure cartilage cells at 20% of the above mentioned concentration (1.0 x 10(7)/ml) were used as the low concentration cartilage cell control group. Samples were collected 8 weeks later. General observations, wet weight, glycosaminoglycans (GAGs) determination and histological and immunohistochemistry examinations were performed.

RESULTSThe expression of type II collagen, type II collagen and aggrecan mRNA were positive in induced BMSCs. In the co-cultured group and the positive control group, pure mature cartilage was formed after 8 weeks of culture in vitro, and the size and shape of the scaffold were maintained. The newly formed cartilage in the two groups were almost the same in appearance and histological properties. The immunohistochemistry results indicated that the cartilage cells of the two groups all expressed ample cartilage-specific type II collagen. The average wet weight and GAG content in the co-cultured group reached more than 70% of those in positive control group. Only an extremely small amount of immature cartilage tissues was formed in local regions in pure BMSC group, and the scaffold was obviously shrunk and deformed. Although the wet weight of newly generated cartilage tissue in the low concentration cartilage cell group reached 30% of that in positive control group, the scaffold was obviously shrunken and deformed. Only regional and discontinuous cartilage tissues were formed, and the amount of newly formed cartilage was obviously less than that in the co-culture group and the positive control group.

CONCLUSIONSChondrocytes can provide a micro-environment for the formation of cartilage, and also effectively induce BMSC to differentiate into chondrocytes and form tissue-engineered cartilage in vitro.

Aggrecans ; metabolism ; Animals ; Cell Differentiation ; Cells, Cultured ; Chondrocytes ; cytology ; Coculture Techniques ; Collagen Type II ; metabolism ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Swine ; Tissue Scaffolds

OBJECTIVETo study the pattern of polymorphism expression of heat-shock protein 70 (HSP70) family in A549 cell line treated with different concentrations of benzo(a)pyrene (BaP) and its probable biological effect.

METHODTwo-dimensional polyacrylamide gel electrophoresis (2D-PAGE) was used for the HSP70 expression analysis.

RESULTS2D-PAGE showed that when A549 cells were exposed to different concentrations of BaP (0.1, 1.0, 5.0, 10.0 micromol/L) for 24, 48 h respectively, the HSP72 in A549 gradually declined as BaP concentrations increased [the integral OD (IOD)] for 24 h were: 150.36 +/- 26.03, 98.57 +/- 13.34, 64.92 +/- 15.03, 34.65 +/- 19.10, 32.92 +/- 18.71 respectively, for 48 h: 126.85 +/- 17.41, 106.19 +/- 15.32, 73.64 +/- 21.02, 35.18 +/- 11.95, 16.27 +/- 9.35 respectively), while the IOD of HSP73 did not show any remarkable change (24 h: 102.29 +/- 21.24, 87.71 +/- 18.70, 71.19 +/- 14.08, 71.87 +/- 15.16, 72.78 +/- 17.31 respectively; 48 h: 86.66 +/- 16.86, 75.67 +/- 10.61, 66.83 +/- 12.63, 67.29 +/- 10.26, 91.37 +/- 13.68 respectively).

CONCLUSIONBaP can inhibit HSP72 expression and with certain dose-effect relationship, but cannot affect HSP73 expression.

Benzo(a)pyrene ; Cell Line, Tumor ; Dose-Response Relationship, Drug ; Electrophoresis, Polyacrylamide Gel ; HSP70 Heat-Shock Proteins ; genetics ; metabolism ; Humans ; Polymorphism, Genetic

OBJECTIVETo explore the methylation status of 5' CpG island of fragile histidine triad (FHIT) gene in plasma and the expression of FHIT protein in cancer tissue of cervical cancer patients.

METHODSMethylation-specific PCR (MS-PCR) was employed to examine methylation of FHIT gene in 151 plasma samples before treatment. The immunohistochemistry was used to the expression of FHIT protein in cancer tissues.

RESULTSCpG island methylation of FHIT was detected in 31.13% of plasma samples. The expression of FHIT protein was decreased or discarded in 59.60% of cervical cancer tissues. Among them 47.78% was included in methylation positive samples.

CONCLUSIONCpG island methylation of FHIT gene in plasma plays an important role on cervical cancer, which results in decreased expression of FHIT protein. It can be used to diagnose and evaluate the effect of treatment to cervical cancers.

Acid Anhydride Hydrolases ; blood ; genetics ; metabolism ; Adult ; Aged ; DNA Methylation ; Female ; Humans ; Immunohistochemistry ; Middle Aged ; Neoplasm Proteins ; blood ; genetics ; metabolism ; Polymerase Chain Reaction ; Uterine Cervical Neoplasms ; blood ; genetics ; metabolism

Objective To study the changing pattern of infant mortality and under-5 mortality rate in China from 2000 to 2006, and to evaluate China's progress in achieving the United Nations' Millennium Development Goal 4. Methods A population-based survey was conducted through a nationwide multi-level surveillance network. The mortality rate and the proportion of death for children under 5 were analyzed. Results The infant mortality rate (IMR), under-5 mortality rate (U5MR) in China dropped to 17.2, 20.6 per 1000 live births in 2006, respectively, comparing to 32.2 and 39.7 per 1000 live births in 2000. In urban areas, IMR, U5MR dropped to 8.0, 9.6 per 1000 live births in 2006, respectively while they were 11.8 and 13.8 per 1000 live births respectively in 2000. In rural areas, IMR, USMR dropped to 19.7 and 23.6 per 1000 live births in 2006, respectively but they were 37.0 and 45.7 per 1000 live births respectively in 2000. During this period, the mortality rates due to pneumonia and diarrhea had dropped sharply. The proportion of deaths due to pneumonia, diarrhea also dropped from 19.5%, 4.9% in 2000 to 15.6%, 3.7% in 2006, respectively. In urban areas, the proportion of deaths due to pneumonia dropped from 9.9% in 2000 to 9.8% in 2006, In rural areas, the proportion of deaths due to pneumonia, diarrhea dropped from 20.1%, 5.2% in 2000 to 16.2%, 4.0% in 2006, respectively. Conclusion The U5MR in China remarkably dropped from 2000 to 2006. Based on data through the surveillance program, China should be able to accomplish the Millennium Development Goals 4 of the United Nations as planned.

Objective To assess the changes and the leading cause of deaths for children under 5 years old,in China,during 2000-2010,with the aim of evaluation on the progress in achieving the relative goal set by "National Program of Action for Child Development in China (2001-2010)",and understanding the related challenges.Methods Data used in this study were collected from the population-based National Maternal and Child' s Health Surveillance Network of China.Infant Mortality Rate (IMR),Under-5-mortality rate (U5MR) and the leading cause of deaths for under-5 children were analyzed.Results Nationwide IMR and U5MR in 2010 dropped by 59.3％and 58.7％ respectively,compared to that in 2000.Decreases by 50.8％ and 47.1％ in IMR and U5MR were observed in urban areas,and 56.5％ and 56.0％ in rural areas during this period.Compared with data from 2000,the leading causes-specific U5MR in 2010 had significantly declined.The top 5 leading causes of death in 2010 were premature birth/low birth weight,pneumonia,birth asphyxia,congenital heart disease and accidental suffocation,but were different in urban and rural areas.In 2010,both IMR and U5MR from the rural areas were 2.8-folds than that of the urban areas.In addition,IMRs in the Middle and Western parts of China were 1.5 and 2.3-folds respectively of that in the East,and U5MR in Middle and West was 1.5 and 2.2-folds respectively of that in East.Conclusion IMR,U5MR and the leading causes specific mortality rate in China declined remarkably from 2000 to 2010,and the goal set by "National Program of Achon for Child Development in China (2001-2010)" had been successfully achieved.However,the disparity on child' s health in regions and in urban or rural areas,still remained a challenge.

Objective: To investigate the surgical methods and outcomes of the enlarged translabyrinthine approach in the removal of large acoustic neuromas. Methods: A large mastoidectomy involved complete exposure of the sigmoid sinus, the dura behind the sinus for at least 1 cm, the superior petrosal sinus and the middle fossa dura. The jugular bulb was exposed and pressed downwards if necessary. The internal auditory meatus was skeletonized and uncovered for at least 270°.The debulking of the tumor began inside the anterior and inferior poles in order to find the brainstem and the facial nerve root as early as possible, and then the dissection of the nerve was done medially to laterally. Intraoperative facial nerve monitoring and postoperative CT and MRI were done in all cases. Results: Total removal was achieved in all 18 patients with tumors larger than 3 cm (mean size: 4.2 cm). There were no deaths or other complications such as intracranial infection and persistent cerebrospinal fluid leakage. There were no obvious cerebral sequelae. The facial nerve was preserved both anatomically and functionally in 14 cases, with Grade Ⅰ or Ⅱ in 8 cases, Grade Ⅲ or Ⅳ in 6 cases. Nerve interruption occurred in 4 patients who all had severe facial palsy or nerve interruption before operation. Sixteen patients resumed work within 1-3 months. Conclusion: Total removal of large acoustic neuroma could be acomplished via the translabyrinthine approach, with good preservation of facial nerve function and minimum incidence of morbidity.

OBJECTIVETo explore heat shock protein 70 (HSP70) expression of A549 cells and its role in DNA damage caused by benzo(a)pyrene (BaP).

METHODSHuman adenocarcinoma A549 cells were cultured in vitro, exposed by different concentrations of BaP (0, 1.25, 2.50, 5.00, 10.00 micro mol/L) for 6 hours, or 10 micro mol/L of BaP for different time (0, 4, 8, 12, 16, 24 and 48 h). Then HSP70 expression and DNA damage were detected using Western-blot and single cell gel electrophoresis (SCGE) assay respectively, and the relationship between HSP70 expression and DNA damage was further analyzed.

RESULTSThe integral optical densities of HSP70 in A549 cells treated with 1.25, 2.50, 5.00 and 10.00 micro mol/L BaP for 6 h (49.63 +/- 1.30, 45.72 +/- 1.03, 40.53 +/- 0.95, 37.50 +/- 1.20 respectively) were lower than that of the control cells (59.43 +/- 1.17) (P < 0.05). When A549 cells were exposed to 10 micro mol/L BaP for 4, 8, 12, 16 h, the integral optical densities of HSP70 were 33.33 +/- 0.80, 29.23 +/- 0.91, 12.51 +/- 0.96, 9.50 +/- 1.25 respectively, and there was an increasing tendency of the expression of HSP70 for 24 - 48 h (20.06 +/- 1.38, 24.51 +/- 1.39), however, all were different from that in control group (56.59 +/- 0.85) (P < 0.05). DNA damage scores in 10(6) A549 cells treated with 2.50, 5.00 and 10.00 micro mol/L BaP for 6 h (23,718 +/- 2,938, 30,128 +/- 2,937, 44,231 +/- 3,846) were significantly higher than that of the control cell (9,615 +/- 1,923) (P < 0.05). When A549 cells were exposed to 10 micro mol/L BaP for 4, 8, 12, 16, 24, 48 h, DNA damage scores (16,667 +/- 4,003, 38,461 +/- 1,924, 5,615 +/- 3,847, 76,282 +/- 2,937, 7,513 +/- 1,110 and 58,975 +/- 9,487) were also higher than that of control group (P < 0.05). There was a negative correlation between DNA damage and the expression of HSP70 when A549 cells were exposed to different concentrations of BaP.

CONCLUSIONHSP70 might enhance intracellular defenses against DNA damage induced by BaP.

Benzo(a)pyrene ; toxicity ; Blotting, Western ; Cell Line, Tumor ; Comet Assay ; DNA ; drug effects ; genetics ; DNA Damage ; Dose-Response Relationship, Drug ; HSP70 Heat-Shock Proteins ; analysis ; Humans ; Time Factors

This study was aimed to investigate the expression level of murine double minute 4 (MDM4) mRNA in chronic lymphocytic leukemia (CLL) and its prognostic value in CLL. By means of β-actin as internal reference, the real-time quantitative RT-PCR was set up. The expression of MDM4 mRNA in 66 CLL patients was measured by fluorescence dye SYBR Green I. The dispersion of MDM4 expression ratio of groups with different prognostic factors was described by using Mann-Whitney U test. The results showed that the median MDM4 mRNA expression level was 0.037098 (0.088245-0.014875) in CLL patients. The expression level of MDM4 mRNA was significantly higher in patients with P53 gene deletion than that in patients without P53 gene deletion (0.13167 vs 0.030927) (p < 0.001), and also significantly higher in patients with P53 mutation than that in patients without P53 mutation (0.13167 vs 0.03077) (p < 0.001). MDM4 expression was also associated with Binet stages (p = 0.044) and ATM gene deletion (p = 0.046), but was not associated with LDH (p = 0.216), β(2)-MG (p = 0.314), TK1 (p = 0.300), ZAP-70 (p = 0.559), CD38 (p = 0.513) and IgVH mutation status (p = 0.333). It is concluded that the expression level of MDM4 is significantly higher in patients with P53 deletion or mutation. MDM4 expression is significantly associated with Binet stages and ATM gene deletion. MDM4 may be an important prognostic factor in CLL.
Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Leukemia, Lymphocytic, Chronic, B-Cell ; diagnosis ; genetics ; metabolism ; Male ; Middle Aged ; Nuclear Proteins ; genetics ; metabolism ; Prognosis ; Proto-Oncogene Proteins ; genetics ; metabolism ; RNA, Messenger ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Tumor Suppressor Protein p53 ; genetics

BACKGROUNDComputer-assisted procedures have recently been introduced for navigated femoral neck screw placement. Currently there is little information available regarding accuracy and efficiency of the different navigated procedures. The aim of this study was to compare two fluoroscopic navigation tracking technologies, a novel bi-planar robot navigation and standardized optoelectronic navigation, versus standard freehand fluoroscopic insertion in a Synbone hip model.

METHODSEighteen fixed Synbone hip models were divided into 3 groups. C-arm navigated cannulated screws (AO-ASIF, diameter 7.3 mm) were inserted using freehand targeting (control group). A novel bi-planar robot system (TINAV, GD2000) and an optoelectronic system (Stryker OTS Navigation System) were used for the navigated procedures (robot group and optoelectronic group). Accuracy was measured using radiographic evaluation including the measurement of screw parallelism and decentralization, and joint penetration. To evaluate the efficiency, the number of guidewire passes, operative time and fluoroscopic images taken were noted.

RESULTSThe two computer-assisted systems provided significantly improved accuracy compared to the freehand technique. Each of the parameters, including guidewire passes and number of fluoroscopy images, was significantly lower when using the computer-assisted systems than for freehand-unguided insertion (P <0.05), but operative time was significantly shorter when using freehand-unguided insertion than for the computer-assisted systems (P <0.05). Accuracy, operative time and number of fluoroscopy images taken were similar among the two navigated groups (P >0.05), but guidewire passes in the robot group were significantly less than in the optoelectronic group (P <0.05).

CONCLUSIONSBoth bi-planar robot navigation and optoelectronic navigation were similarly accurate and have the potential to improve accuracy and reduce radiation for freehand fluoroscopic targeting for insertion of cannulated screws in femoral neck fractures. Guidewire passes in the robot group were significantly less than in the optoelectronic group. However, both navigated procedures were associated with time-consuming registration and high rates of failed matching procedures.

Bone Screws ; Femoral Neck Fractures ; surgery ; Hip ; diagnostic imaging ; surgery ; Humans ; Radiography ; Surgery, Computer-Assisted ; methods

OBJECTIVETo analyze the prognostic factors for chronic lymphocytic leukemia (CLL) with typical and atypical immunophenotype. The parameters analyzed included sex, age, Binet stages, absolute lymphocyte count (ALC), immunoglobulin heavy-chain variable region (IgVH) gene mutation status, ZAP-70 protein, CD38 expression and cytogenetic aberrations.

METHODSAccording to the clinical guideline and scoring system for CLL in Britain, among 77 patients, 61 patients with score 5 called typical immunophenotype CLL, 16 with score 4 or 3 were atypical immunophenotype CLL. Multiparameter flow cytometry was employed for immunophenotypic analysis in 77 CLL patients for CD5, CD19, CD23, FMC7, sIg, CD20, CD79b expression and ZAP-70 protein and CD38. IgVH mutation status was detected by multiplex RT-PCR and sequencing of the purified PCR amplification products. Fluorescence in situ hybridization (FISH) and a panel of probes were used to detect cytogenetic aberrations.

RESULTSThere was no significant difference between the two groups in sex, age, ZAP-70 and IgVH mutation status (P=0.398, P=0.189, P=0.268 and P=0.131, respectively). The incidence of ALC> or =50 x 10(9)/L, Binet B + C, CD38> or =30% in atypical CLL patients (43.8%, 87.5% and 43.8%, respectively) were higher than that in typical group (16.4%, 36.1% and 16.4%, respectively) (P=0.026, P<0.01 and P=0.026, respectively). The proportion of typical patients (26.8%) with a 13q14 deletion as sole abnormality was higher than that of atypical patients (7.6%), and that with deletion of 11q22 or 17p13 was lower than that of atypical patients (12.2% vs 46.2%) (P=0.022).

CONCLUSIONThere were obvious differences between the typical immunophenotype CLL and atypical CLL in ALC, Binet stages, CD38 expression level and cytogenetic aberrations.

ADP-ribosyl Cyclase 1 ; metabolism ; Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Immunoglobulin Heavy Chains ; genetics ; Immunoglobulin Variable Region ; genetics ; Immunophenotyping ; Leukemia, Lymphocytic, Chronic, B-Cell ; genetics ; immunology ; metabolism ; Male ; Middle Aged ; Mutation ; Neoplasm Staging ; Prognosis ; ZAP-70 Protein-Tyrosine Kinase ; metabolism