Purpose:To study whether the inhibition of phosphatidylinositol 3-kinase (PI3K)/Akt survival pathway could sensitize the response of tumor cells to some chemotherapyeutic agents. Methods:The different tumor cells has been treated with the combination of isoform specific Akt inhibitor and either adriamycin or camptothecin; the quantitation of the induction of apoptosis by drugs has been estimated with caspase 3 assay; immunoprecipitation western blotting has been used to evaluate the inhibition of the phosphorylation of different isoforms of Akt after the treatment. Results:①The inhibitors could reduce the phosphorylation of Threonine 308 and Serine 473 of isoform specific Akt. ②The inhibition of any one isoform specific Akt could not reverse the resistance of tumor cells tested to chemotherapeutic drugs, but it is not the same case if blocking of two isoforms of both Akt1 and Akt2 was done at the same time. ③The synergistic effects of Akt inhibitors is maybe relative to the level of endogenous PTEN(phosphatase and tensin homolog deleted on chromosome 10) expression. Conclusions:It is required to inhibit two isoforms of both Akt1 and Akt2 in order to maximally sensitize the tumor cells to chemotherapy.

BACKGROUND: The implanted cartilage calls can synthesize cartilage matrix as cartilage in cartilage tissue enginsedng, and the density of implanted cells is the key point.OBJECTIVE: To evaluate the effect of cell seeding concentration on the chondrogenic differentiation of the adipose dadved sromal cells (ADSCs).DESIGN, TIME AND SETTING: The in vitro cellular-scaffold observation was performed at the cytobiological laboratory of China Medical University from November 2007 to July 2008.MATERIALS: Six male SD rata with clean grade were supplied by the Experimental Animal Center of China Medical University.METHODS: Totally 5 g/L type ; collagen solution and 20 g/L chitosan was mixed in a mould with volume ratio of 7:3, after lyophillization, it was cut into pieces with 5 mm ～ 5 mm x 2 mm, followed by crosslinking with ethanol contained of 2% chondroitic acid at room temperature. After washing with double distilled water and freeze drying, the chitosan-collagen-chondroitin sulfate copolymar matrices scaffolds were harvested. ADSCs isolated from rat inguinal fat pads were digested with collagenase and trypsase. The prepared scaffolds were randomly divided into 3 groups, and the third passage cells with density of 2×10 9/L,2×10 109/L, and 2×10 11/L were seeded into chitosan-coflagen-chondroitin sulfate scaffolds, and cultured in chondrogenic medium for 3 weeks.MAIN OUTCOME MEASURES: The expression of cartilage specificity gene was detected by hematoxylin-eosin staining, type Ⅱ collagen immunohistochemical staining and RT-PCR.RESULTS: Hematoxylin-eosin staining showed that after 3 weeks of culture, the cell proliferated and differentiated well, especially in 2x101～/L group, more extrocelluer matrices were produced and cartilage lacuna-structure could be seen. The type Ⅱ collagen was positive expressed in each group, which showed a gradually increasing tendency with the cell seeding concentration increasing. RT-PCR showed that the expression of proteoglycen and type Ⅱ collagen mRNA were slowly increased. However the expression of Ⅹ collagen mRNA was decreased with increasing cell seeding concentration.CONCLUSION: The chitosan-collagen-chondroitin sulfate copolymer matrices can provide an appropdate environment for the generation of cartilage-like tissues and high call seeding concentration of 2×1010/L facilitate ADSCs to differentiate into cartilage.

Objective To evaluate the character of the collagen-chitosan-chondroitin sulfate scaffold seeded with rat adipose tissue-derived stromal cells. Methods A dipose tissue were harvested from 6 weeks old Wistar rats and the stromal cells were harvested by type Ⅰ collagenase and then cultured in vitro. Type Ⅰ collagen was fully mixed with chitosan, freeze-dried and cross-linked with chondroitin sulfate, then freeze-dried again and sterilized by ethylene oxide. The pore diameter, water content, porosity of the scaffold were tested. The adipose tissue-derived stromal cells were digested, seeded into the plates, scaffold, and cen-trifuged into pellet, and then induced into cartilage. MTT detection for cell proliferation was done. After 3 weeks, the cell morphology, and cell proliferation and adhesion were observed, and chondrngenic differenti-ation was also analyzed. Results The pore diameter, water content, porosity tested for the scaffold showed an appropriate form. Cell proliferation showed faster in the scaffold and pellet culture system after 5 day, there was still cell proliferation in the scaffold system after 14 days but no obvious changes in the pellet cul-ture system; ceils on the scaffold proliferated densely showed by histological staining, but there was a scaf-fold structure residues in the inner layer. The finding of type Ⅱ immunohistochemistry stain showed that cells express strong positive for type Ⅱ collagen in the scaffold and pellet culture system whereas it was weakly positive in the plate culture system; the specific mRNA for cartilage, type Ⅱ collagen, aggrecan and SOX-9 were expressed in all three systems showed by RT-PCR, but type X collagen was expressed continu-ously in the plate culture system and expressed after 21 days in the pellet culture system, whereas it was not detected in the collagen-chitosan-chondroitin sulfate scaffold system. Conclusion The parameters of the collagen-chitosan-chondroitin sulfate scaffold were suitable in our study. The results suggested that it can promote the adipose tissue-derived stromal cells proliferation and chondrogenic differentiation better than the plate and pellet culture systems and maintain the phenotype of chondrocytes well; it is the optimal choice for cartilage tissue engineering in the future.

Objective:To study the clinical significance of β2-microglobin(β2-MG)and fibronection (Fn) contents in patients with nasopharyngeal carcinoma.Methods:The contents of serum β2-MG and Fn were detected by radioimmunoassay and agar-diffusion methods in 60 patients with nasopharyngeal carcinoma before and after radiotherapy,also in 60 normal subjects as control group.Results:The contents of serum β2-MG and Fn were 2.99±1.11mg/L(β2-MG)and 134.60±28.93mg/L(Fn) in 60 patients with nasopharyngeal carcinoma,2.16±0.50mg/L(β2-MG)and 196.16±34.65mg/L(Fn)in 60 health subjects,respectively.Those results showed that the content of β2-MG in patients group was higher than that in control group (P＜0.05),and the content of Fn in patients group was lower than that in control group(P＜0.05).After radiotherapy of patients group,the content of β2-MG were lower(2.16±0.55mg/L)than that before radiotherapy(P＜0.05).Conversely,the content of Fn was higher(180.53±34.66mg/L)than after radiotherapy(P＜0.05).Conclusions:The detection of serum β2-MG and Fn have clinical significance on the evaluation prognosis of patients with nasopharyngeal carcinoma.

Background and purpose: The prognosis of completely resected stage ⅢA(N2) non-small cell lung cancer (NSCLC) remains a significant concern. The 5-year overall survival (OS) rates range from 10% to 30%. This study aimed to analyze the patterns of first failure in completely resected stage ⅢA(N2) NSCLC and to assess the actuarial risk of developing metastasis at different sites and to guild standard clinical practice. Methods: Patients withⅢA(N2) NSCLC who had undergone radical surgery in our hospital from Jan. 2005 to Jul. 2012 were retrospectively reviewed. The progression-free survival (PFS), the OS, patterns of first failure, the actuarial risk were analyzed. The cumulative incidence of first failure was determined using the Kaplan-Meier analysis. Results: Among 357 patients who met the eligibility criteria with completely resected stage ⅢA(N2) NSCLC, 5-year OS was 36.9%. There were 284 (77.6%) patients experiencing disease failure: 61 with local failure, 197 with local and distant failures, and 26 patients with local recurrence as the first failure. Brain, bone and lung were the main sites of distant failure as the first failure, while brain was the most common site. There were 67 patients developing brain metastases (BM) as the first site of failure. The median time of local failure as the first site of failure was 13.6 months, and the time to develop distant recurrence was 15.1 months. 92.5% BM developed in 3 years after the complete resection. Conclusion: As the first failure, the rate of distant failure was much higher than that of local failure in completely resected stage ⅢA(N2) NSCLC. Brain was the most common site of distant failure as the first failure. These results can be helpful in guiding standard clinical practice and evaluating the outcome of comprehensive treatment.

Objective:To investigate the role of soluble Fas ligand (sFasL) in the mechanism of immune escape and counterattack of colorectal cancer.Methods:ELISA was used to detect the level of sFasL in the sera of colorectal cancer patients and the supernatant of SW480. Transmission electron microscopy (TEM), flow cytometry (FCM) analysis and fluorescence microscopy were used to examine the apoptosis of Jurkat induced by the sera of colorectal cancer patients and the supernatant of SW480. The apoptosis was also studied after pretreatment of Jurkat by Fas blocking antibody ZB4. The supernatant of African green monkey cell line Vero was used as control.Results:The concentration of sFasL in sera of colorectal cancer patients was 12.21?1.14 ?g/L before treatment, the concentration decreased significantly after treatment(P

Objective The aim of this study was to evaluate the outcomes of auditory perception skills after cochlear implantation in children with malformed inner ear and compare them with a group of congenitally deaf chil ‐dren implantees with a normal inner ear .Methods 21 children with inner ear malformation were retrospective stud‐ied .There were 9 cases with large vestibular aqueduct syndrome (LVAS) ,7 with Mondini abnormality ,5 with com‐mon cavity .The postoperative outcomes of these 21 cases were compared with 21 cases with normal inner ear struc‐ture .The outcomes of all the children after the surgery in 1 year were studied using the soundfield test in frequency ranging 0 .5 to 4 kHz and the auditory perception skills ,and the auditory perception skills consisted of natural envi‐ronmental sound recognition ,consonants recognition ,vowels recognition ,numeral recognition ,tone recognition , monosyllabe recognition ,disyllabe recognition ,trisyllabe recognition ,short sentences recognition ,selective hearing recognition .Results The results of soundfield test and auditory perception skills after cochlear implantation in 9 children with LVAS and 6 with Mondini abnormality had no significant difference (P > 0 .05) comparing with the control cases .Postoperative thresholds in soundfield test were 50 ~ 75 dB HL for 1 case with severe Mondini abnor‐mality ,the mean value of hearing ability score was 70 .5% ,and less than the results of control cases .Postoperative mean thresholds in soundfield test were 65 .26 ± 5 .13 dB HL for 5 cases with common cavity ,the hearing ability score was less than the results of control cases (P< 0 .05) .Conclusion The effect of rehabilitation had no difference between the children with LVAS and the cases with normal inner structure after cochlear implantation ,but was poo‐rer in children with severe Mondini abnormality and common cavity .It was necessary to evaluate the degree of mal‐formation of inner ear structure before cochlear implantation .

Objective The purpose of this study was to investigate the correlation of high sensitivity C-reactive protein (hsCRP) and fibrinogen with carotid artery arteriosclerosis of patients with cerebral infarction. Methods One hundred and thirteen patients with cerebral infarction were assigned as study group, and 102 healthy persons as control group. The levels of serum hsCRP and Fib in the two groups were measured. The carotid artery arteriosclerosis and carotid intimal-medial thickness (IMT) were examined by color Doppler and B-ultrasound. Results The value of IMT between study group and control group was statistically significant. The positive rates of carotid artery arteriosclerosis plaque and vulnerable plaque in study group were significantly higher than those in control group (all<0.05) . The level of serum hsCRP was significantly higher in study group than that of control ( <0.05) . The level of serum Fib between study group and control group was not statistically significant ( >0.05) . Conclusion The level of hsCRP was closely related to the degree of carotid artery arteriosclerosis and the occurrence and development of cerebral infarction. But the level of Fib was not closely related to the degree of carotid artery arteriosclerosis.

Double-hit lymphoma (DHL) is a kind of disease with features intermediated between diffuse large B-cell lymphoma (DLBCL) and Burkitt lymphoma (BL),usually accompanied by myc gene breakpoint with other recurrent chromosomal breakpoint and mainly involving myc and bcl-2 translocation.The presentation of this disease is characterized by elevated serum lactate dehydrogenase levels,B symptoms,bone marrow involvement,advanced stage disease,extranodal involvement,and central nervous system involvement.Because its features are similar with DLBCL and BL,it's difficult to distinguish them by pathological diagnosis.At present,the differential diagnosis is mainly by chromosomal analysis (G-banding),FISH and immunohistochemistry.This subtype received a poor response to conventional chemotherapy for DLBCL,and has a poor prognosis.The median survival time is only 0.2-1.5 years.Currently,the main regimens include RCHOP,RICE,RCVD,methotrexate prophylaxis for central nervous system involvement,high-dose chemotherapy and bone marrow transplantation.

OBJECTIVE:To establish a method for the simultaneous determination of acetic acid and succinic acid in Banxia syrup. METHODS:RP-HPLC was performed on the column of GL InterSustain-C18 with mobile phase of 0.03 mol/L ammonium di-hydrogen phosphate buffer solution(pH2.0)-methanol(gradient elution)at a flow rate of 0.8 ml/min,the detection wavelength was 210 nm,column temperature was 30 ℃,injection volume was 10 μl. RESULTS:The linear range was 0.020 98-0.209 8 μg/ml for acetic acid(0.999 9)and 12.04-120.4 μg/ml for succinic acid(r=0.999 9);limits of quantification were 0.15,18.24 ng,limits of detection were 0.045, 5.53 ng;RSDs of precision, stability and reproducibility tests were lower than 2%;recoveries were 97.45%-101.68%(RSD=1.39%,n=9) and 98.31%-101.08%(RSD=1.01%,n=9). CONCLUSIONS:The method is accurate and rapid,and suitable for the simultaneous determination of acetic acid and succinic acid in Banxia syrup.