AIM: To explore whether human telomerase reverse transcriptase (hTERT) antisense oligodeoxynucleotide (ASODN) could enhance the sensitivity of acute lymphoblastic leukemia cells from relapse patients to cisplatin. METHODS: The expression levels of hTERT protein were detected by immunofluorescence using fluorescence isothiocyanate (FITC), the number of viable cells was determined using the trypan blue dye exclusion assay, and apoptosis was detected by morphological observation and flow cytometric cell cycle analysis. RESULTS: The expression of hTERT protein was inhibited after treatment with hTERT ASODN. Treatment with cisplatin combined with hTERT ASODN had significantly reduced the number of viable acute lymphoblastic leukemic (ALL) cells (P<0.05). In morphological observation of apoptotic cells using Hoechst33258 and PI double staining techniques, cells displayed classic apoptotic changes in the presence of cisplatin or cisplatin combined with hTERT ASODN or ASODN at 48 h. Apoptotic rates of cells treated with cisplatin and ASODN were higher than that of cells treated with cisplatin alone (P<0.05). CONCLUSION: hTERT ASODN could increase sensitivity of cultured primary acute lymphoblastic leukemia cells from relapse patients to cisplatin.

AIM: To explore the delayed protection of heme oxygenase-1 (HO-1) in the exercise preconditioning (EP) from the myocardial relative ischemia reperfusion injury (rI/R). METHODS: 40 Wistar Rats were divided into 5 groups randomly: control group (CN), rI/R group (IR), EP+rI/R group (EI), HO-1 inductor hemin+rI/R group (HE) and HO-1 inhibitor ZnPP+EP+rI/R group (EZ). The following indexes were detected, including the HO-1activity in myocardium, the cardiac function parameter-pressure-rate product (heart rate × left ventricular developed pressure, PRP) and the content of MDA in coronary effluent. RESULTS: After myocardial rI/R, HO-1 activity increased significantly. Moreover, EP or HO-1 inductor could enhance this effect manifestly. Nevertheless, when the HO-1 inhibitor was administered before EP,HO-1 activity decreased. In addition, there was no distinct difference in the HO-1 activity between EI group and HE group. At the 30 min point of reperfusion, the PRP recovery rate of EI group was higher clearly than that of IR group. However, there was reverse effect between the EZ group and the EI group. The MDA in coronary effluent of EI group, EZ group and HE group were lower obviously than that of IR group and there was significant difference between EI group and EZ group. CONCLUSION: EP could protect the heart from the rI/R injury occurring 24 hours later, which might be performed through activating the HO-1.

AIM: To investigate the potential of murine epidermal stem cell (ESC) differentiation after seeded in a biodegradable carrier and implanted subcutaneously into syngeneic recipient mice. METHODS: ES cells were induced in vitro to differentiate into ESCs. After stained with a fluorescent dye Hoechst 33342, these ESCs were seeded into a polyglycolic acid (PGA) net containing collagen gel, functioning as a cell carrier, and implanted subcutaneously into 129/J mice, which were syngeneic to these stem cells. RESULTS: The ESCs kept alive in the implant when observed under a fluorescent microscopy 3 weeks or longer after implantation, and could differentiate into hair follicle - like structure,glandular structure, and gave rise to additional structures displaying features resembling native dermis. No apparent rejection or severe side effects were observed at least 10 weeks post- implantation. CONCLUSION: It is feasible to use these ESCs as seed cells in the study to fabricate dermal equivalent having the potential to develop dermal appendages.

AIM: To investigate the feasibility and its mechanisms of improving therapeutic effect by antisense gene therapy combined with chemotherapy in osteosarcoma. METHODS: The human osteosarcoma implanted tumor model in the nude mice was established. By intratumoral injection and abdominal cavity administration, the tumor bearing mice were treated with survivin ASODN in combination with diamminedichloroplatinum (DDP) for a week. Comparison with each single - agent therapy and control group was performed in aspects such as tumor growth condition, pathological changes of tumor tissues; survivin protein expression in tumor tissues by immunohistochemistry, survivin mRNA expression levels by RT -PCR method and tumor apoptosis by Tdt -mediated dUTP nick end labeling (TUNEL). RESULTS: All nude mice survived the therapy. As compared with the control group, the antisense gene therapy group presented synchronous decrease in survivin mRNA and protein expression; all therapy group displayed tumor growth inhibition and cell apoptosis with different extent; while in contrast to single - agent therapy group, the combined therapy group showed stronger inhibition of tumor growth and abundant tumor cell apoptosis with the highest apoptotic rate. CONCLUSION: Synergistic effect was achieved by combination of DDP with ASODN that may overcome drug resistant of DDP and the combined strategy may shed new light on the cancer therapy.

AIM: To investigate the cytological basis and differentiating conditions of human bone marrow mesenchymal stem cells (hMSCs) differentiated into cells of the endothelial lineage in vitro. METHODS: hMSCs were isolated by density gradient centrifugation and fractionated on a 1 073 g/L Percoll. The combination of VEGF165 and various matrix proteins including fibronectin (FN) and type I collagen (Col) was used to induce hMSCs in vitro. Cells were characterized by immunohistochemistry, cytochemistry, FACS and ultrastructure to identify and detect the differentiated population and markers. RESULTS: hMSCs was positive for KDR. PAS reaction was positive and ultrastructure of hMSCs showed glycogen- pool in ectoplasm. Glycogen reducing or disappear suggested that stem cells have occurred differentiation. Induction of hMSCs resulted in the increase of KDR, β1 integrin and CD34. CONCLUSION: hMSCs were induced to a transit population (TP( )) that differentiated toward the endothelial progenitor cells ( EPC), but not a really EPC. hMSCs pedigree diagram of differentiation was hMSCs→TP→EPC→endothelial cells (Ecs).

AIM: To evaluate the influences of triptolide on serum cytokines, symptoms and pulmonary function in patients with steroid - resistant asthma, so as to investigate if there is therapeutic effect of triptolide on these patients. METHODS: Sixteen patients with steroid - resistant asthma were randomly divided into two groups (A and B, n =8 for each group). All of the patients took procaterol (50 - 100 μg/d) and theophylline (400 mg/d) orally as baseline treatment. Additionally, triptolide was prescribed for group A (33 μg, orally, three times per day for 4 weeks). Asthmatic symptom score calculation, serum cytokines ( interferon - γ, IFN - γ; interleukin - 4, IL - 4; and interleukin - 5, IL - 5)determination and pulmonary function test (FVC%, FEV1%, PEF%, V50% and V25% ) were undertaken before and at the end of the study. RESULTS: At baseline, no significant difference was found between group A and group B with respect to the above mentioned indices. Following the administration of triptolide, group A had significantly increased serum IFN -γlevel, FVC%, FEV1%, PEF%, V50% and V25%, and significantly decreased asthmatic symptom score and serum IL-4, IL-5 levels (P＜0.01 compared with baseline in the same group, and P＜0.05 compared with group B at the end of the study). Compared with baseline, no significant change was observed for group B regarding all the indices at the end of the study. CONCLUSION: Triptolide in combination with procaterol and theophylline may be a novel and effective strategy for the treatment of steroid - resistant asthma.

AIM: To observe the inhibitory effect of interferon-α ( IFN-α) on the growth invasiveness and metastasis of human gastric carcinoma cell line BGC-823, and mechanism of its action. METHODS: We detected the influence of IFN-α on the proliferative ability of BGC-823 in cell culture system, the cell vitality with the MTT colorimetric assay, and the cell cycle with flow cytometer (FCM). The regulatory functions of IFN-α to the expression of E-cadherin and matrix metalloproteinase-2 ( MMP-2) in tumor cells were estimated by immunohistochemical analysis ( S-P). The ultrastructural changes of the junction among the tumor cells were observed under electron microscope. RESULTS : IFN-α can significantly inhibit the growth of human gastric carcinoma cell line BGC-823 in a dose-dependent manner. When the concentration of IFN-α was ≥106 U/L, the cell proliferation can be effectively suppressed,the suppression rate was ≥ 12. 2%, and the blockage appeared at the phase of G1-S of the cell cycle. Under the induction of IFN-α, the expression level of the cell E-cadherin increased while the MMP-2 decreased. The changes on ultrastructure of the cells showed the increased adhesive junctions and the relative compact structure. CONCLUSION: IFN-α can suppress the growth of human gastric carcinoma cell line BGC-823 through its influence on cell cycle. IFN-α can regulate the expression of E-cadherin and MMP-2, make the cell junction closely, so that it has the potential on restricting the invasion and metastasis of gastric carcinoma cells.

AIM: To study the basic mechanism of transcriptional regulation, NKX3.1 gene promoter was cloned and its promoter activities in prostate cancer cell lines and other cancer cell lines were tested. METHODS: 1.04 kb - promoter fragment of NKX3. 1 gene was obtained by PCR and cloned into pGL3 - basic and pEGFP - 1 that are promoter - less reporter vectors to examine its promoter activity driving the reporter gene transcription. The promoter activity was determined by dual -luciferase reporter assay and the expression of GFP reporter observed under fluorescence micro scope. RESULTS: The sequence of the cloned 1.04 kb promoter proved to be correct by DNA sequencing. The dual - lu ciferase reporter assay (M1/M2) showed that the promoter activity in LNCaP cell transfected with pGL3 - 1.04 kb promoter was about 1.5 - fold higher than that of pGL3 - control transfection and 50 - fold higher than that of pGL3 - basic transfec tion. To investigate the 1.04 kb - promoter activity in different tumor cell lines, the constructed pGL3 - 1.04 kb promoter and pEGFP - 1.04 kb promoter were transfected into several cell lines, respectively. The results showed that the activity of 1.04 kb promoterin LNCaP was highest among the tested cell lines. Multiple consensus sequence elements have been iden tified within the 1.04 kb fragment using TRANSFAC database. Further experiments will be done to determine their founc tions. CONCLUSION: Cloned 1.04 kb fragment upstream of NKX3.1 gene presented a strong promoter activity and its activity was highest in LNCaP cell among the tested tumor cell lines.

AIM: To explore the effects of hyperbaric oxygen (HBO) therapy on language function of aphasics and it's mechanisms. METHODS: Forty patients with aphasics after mild and moderate cerebral trauma and stroke were chosen and divided into therapy group (20 patients) and control group (20 patients). All the patients in both groups received routine therapy.Besides routine therapy, patients in therapy group also received HBO therapy. The HBO treatment contained three courses, each lasts for 15 days at an interval of 3 days. The language functions ( including 11 sub - items) were tested with the Apparatus ZM2.1for Diagnosis and Treatmern of Language Disorders (Language Disorders ZM2.1) before and after each course of HBO and be compared between the two groups. RESULTS: The therapeutic effects between the therapy group and the control group: the scores of 10 sub- items are significantly higher than those of the control group after the 2nd course (P＜0. 05); After the 3rd course, all 11 sub - items were improved significantly ( P ＜ 0.05). The therapeutic effects before and after each course of the therapy group:the scores of the advanced dictation, voice expression and semantic expression increased significantly after the 1st and the 2nd course ( P ＜ 0.05). CONCLUSIONS: The HBO therapy might facilitate the recovery of the language function of the aphasics by promoting the recovery and the self- reparation of the neurocytes and alleviating the focal ischemia - reperfusion. The result of this effect would facilitate the original recovery velocity and not be focused on some language functions and much more obvious in the 1st and the 2nd course.

AIM: To evaluate the effect of mechanical periodontal treatment combined with tetracycline on per iodontal attachment level (AL) and the avidity of serum antibody against porphyromonas gingivalis( P. gingivalis) in patients with aggressive periodontitis(AgP). METHODS: Twenty- five patients with AgP and twenty periodontally healthy controls were studied (HS). Clinical examination and recordings of AL were performed before and 3,6 and 12 months after the periodontal treatment. The avidity of IgG antibody against P. gingivalis lipopolysaccharide(LPS) was measured by diethylamine dissociation ELISA. RESULTS: A significant improvement in AL was observed following treatment ( P ＜ 0. 01 ). The avidity of serum IgG antibody against P. gingivalis increased compared with controls, and was decreased significantly after mechanical periodontal treatment combined with tetracycline (P ＜ 0. 01 ). CONCLUSION: Ourresults demonstrate that mechanical periodontal treatment combined with tetracycline provides clinically favorable results in patients with AgP.