Objective To investigate the effects of VCAM-1 on migration and invasion of glioma cell lines . Methods The techniques of lentivirus pSGU6/GFP/Neo-based VCAM-1 shRNA and EF1 a-GFP/puro-based VCAM-1 expression vector, the scratch wound healing migration and transwell invasion assays , and the Western blot and cell staining were applied to observe the effects of VCAM-1 expression levels on migration and invasion of glioma cell line cells.There are four groups in T98G cells including control, vector, scramble and shRNA-VCAM-1 groups and three groups in U251 cells covering control, vector and VCAM-1 overexpressed groups ( n=6 per group) .Results The stabled glioma cell lines of T98 G cells with down-regulated VCAM-1 and U251 cells with VCAM-1 overexpression were established by using lentivirus-based VCAM-1 shRNA and expression vector.The ability of scratch wound healing (migration activity) decreased significantly (P<0.01) in T98G cells with lower VCAM-1 expression levels, while the migration activity was obviously improved in U251 cells with overexpressed VCAM-1 ( P <0.05 ) .Similarly, the invasion ability was significantly inhibited ( P <0.05) in T98G cells with silenced VCAM-1, as well as VCAM-1 overexpression could enhance the invasion ability of U251 cells ( P<0.01 ) .Conclusions VCAM-1 improves the migration activity and invasion ability of human glioma cell line cells.

Objective To prepare HCC-targeted lipid ultrasound microbubble containing 10-hydroxycamptothecin (10-HCPT), and to assess its targeting function in vitro. Methods After the biotinylated monoclonal antibody Hab18 was prepared, the biotinylated degree was determined. Microbubbles containing 10-HCPT were prepared by mechanical vibration. Then the biotinylated antibody was attached to the surface of the microbubbles by avidin-biotin interaction. The physical property, entrapment efficiency and the drug-loading amounts of 10-HCPT lipid microbubbles were determined. The combination of biotinylated Hab18 with microbubbles containing 10-HCPT was proved by immunofluorescent assay, and the targeting function of the targeted microbubbles containing 10-HCPT was observed with light microscope, served non-targeted microbubbles as control group. Results About 13 biotin molecules were coupled to each antibody in average. The disposition of the prepared microbubbles was steady and the mean diameter was 1.52 μm. The drug entrapment efficiency was 76.32% and the drug-loading amounts was 21.81%. Red fluorescence was observed at the edge of the microbubbles in immunofluorescent assay, and the conjugation of the targeted microbubbles containing 10-HCPT with 7721 cells was tight while control group was negative. Conclusion HCC-targeted lipid ultrasound microbubbles containing 10-HPTC can be prepared successfully, with higher entrapment efficiency and drug-loading amounts and strong targeting function in vitro.

Objective To up-regulate the expression of Glil gene in periodontal ligament stem cells ( PDLSCs) and to explore the effect of Glil gene on PDLSCs proliferation and osteogenesis differentiation by establishing Glil gene adenovirus vectors. Methods Subcloned Glil to viral backbone vector Adtrack-CMV and transfered the established vector to 293T cells, which was to acquire the virus particles. Trans-fected aim cells,namely PDLSCs,with these virus. Detected its effect on PDLSCs proliferation with CCK-8 assay, and detected the expression of Glil and the bone-related markers ALP and Runx2 through Western blot. Results An adenovirus vector, which were over expressed Glil gene, was successfully constructed and transfected to PDLSCs. Compared with the empty vector group and normal group, the over expressed one had a much slower proliferation rate in CCK-8 assay (P=0. 003). Western blot showed that ALP and Runx2 can be overexpressing os-teogenic differentiated after PDLSCs successfully transfected with the Glil gene. Conclusion Over expressing Glil gene would lead to a much slower proliferation rate in the PDLSCs and an increase of the bone-related markers. It is concluded that Glil can enhance the osteogenic dif-ferentiation capacity in PDLSCs.

Batch and continuous fermentation were adopted to investigate the effect of specific growth rate and amino acid components on RNA accumulation in Candida tropicalis ATCC 20408 in fermentation medium ( FM), yeast peptone dextrose medium (YPD), molasses fermentation medium ( MFM) and FM without corn steep liquor. The data showed that obvious differences in intracellular RNA accumulation were observed at different cell growth phases in bath fermentation prosess, and RNA level reached 11. 8% (g-RNA /g-DCW) during exponential phase, and only 6.9% during stationary phases. It was also found that intracellular RNA accumulation increased with the increase of specific growth rate in continue fermentation prosess, and the highest RNA level reached 15. 6% with the glucose conversion rate of 42. 8% at the dilution rate of 0. 5 h-1. Furthermore, the data showed that RNA lever was notably increased in batch fermentation process when amino acids or peptone was added into the fermentation medium containing no corn steep liquor. Taken together, it was reported for the first time that specific growth rate and amino acid components plays a leading role on the intracellular RNA accumulation in C. tropica lis, and specific growth rate is more important.

Objective To observe the clinical efficacy of cerebral palsy children with liver strong spleen weak syndrome treated with integrated traditional Chinese medicine therapy. Methods Eighty cerebral palsy children with liver strong spleen weak syndrome were divided into treatment group (40 cases) and control group (40 cases). Patients in control group were treated with conventional therapy (including exercise therapy, occupational therapy, guidance education, physical factors, and psychological therapy), and patients in treatment group were treated with integrated traditional Chinese medicine therapy (acupuncture and traditional Chinese medicine medicated bath) additionally. Three months were set for a course of treatment, and the whole treatment lasted for a total of four courses. TCM clinical symptoms were scored every six months, muscle tone (MAS), gross motor function (GMFM), and daily life activity ability (MBI) were evaluated every three months for both groups. Results There were significant differences in the total scores of TCM clinical symptom, MAS, GMFM, and MBI at different treatment time points in the two groups, compared with that before treatment (P<0.01). There were significant differences in the total scores of clinical symptom for Chinese medicine, MAS, GMFM, and MBI at different treatment time points for treatment group, compared with that of control group (P<0.05). Conclusion Integrated traditional Chinese medicine therapy (acupuncture and traditional Chinese medicine medicated bath) for cerebral palsy children on the basis of western medicine therapy can more effectively improve function and life quality of children patients.

Objective To observe the effect of spleen-warming and kidney-tonifying therapy on Toll-like receptor 4 (TLR4) signaling pathway of cerebral infarction sequelae patients. Methods A total of 60 cerebral infarction sequelae patients with spleen and kidney(yang) deficiency were divided into control group and treatment group, 30 cases in each group. The two groups were given function rehabilitation training following by the theory of modern rehabilitation medicine, and additionally the treatment group was treated with spleen-warming and kidney-tonifying herbs orally. The treatment period for the two groups averaged 3 months. Before and after treatment, the scores of traditional Chinese medicine (TCM) symptoms and signs, peripheral blood TLR4 mRNA expression level, and the inflammation-related factors in the two groups were observed, and the observation parameters were compared with 30 healthy volunteers. Meanwhile, the changes of modern rehabilitation evaluation indexes were also evaluated before and after treatment. Results (1) After treatment, the Fugl-Meyer scores for motor function and Modified Barthel Index(MBI) scores for activities of daily living in the treatment group were obviously increased(P < 0.05 or P < 0.01 compared with those before treatment); the MBI scores in the control group was much increased(P < 0.05 compared with those before treatment), but the Fugl -Meyer scores only showed an increasing trend(P > 0.05). (2) After treatment, the predominant symptoms and signs of hemiplegia, facial distortion and dysphasia in the two groups were much improved(P < 0.01 compared with those before treatment), and the improvement in the treatment group was superior to that in the control group(P < 0.05). The control group had no significant effect on relieving the symptoms and signs of dizziness and blurred vision, numbness of limbs, spontaneous sweating, shortness of breath and weakness, and darkish tongue(P > 0.05 compared with those before treatment); the treatment group had obvious effect on the above symptoms and signs, and the effect was superior to that in the control group(P < 0.05 or P < 0.01).(3) For the inflammation-related factors, tumor necrosis factor alpha(TNF-α) level in the treatment group was much decreased and even arrived to the normal level(P > 0.05), and TNF-α level in the control group was decreased but did not arrive to the normal level(P < 0.05); levels of interleukin(IL)-1β, IL-6 and TLR4 mRNA in the two groups were decreased but did not arrive to the normal(P < 0.01), and the decrease in the treatment group was superior to that in the control group(P < 0.05 or P < 0.01). (4) During the treatment period, no obvious adverse effect was found in the two groups. Conclusion Oral use of spleen -warming and kidney –tonifying Chinese medicine combined with modern rehabilitation therapy is effective for the treatment of the sequelae of cerebral infarction patients by relieving the clinical TCM symptoms, and one of the therapeutic mechanisms is probably related with the TLR4 signaling pathway through regulating the expression of inflammatory factors.

Amino Acid Metabolism, Inborn Errors ; diagnosis ; metabolism ; therapy ; Diagnosis, Differential ; Diet ; Glutarates ; metabolism ; urine ; Humans ; Infant ; Male ; Riboflavin ; therapeutic use ; Treatment Outcome

Objective To preparation a kind of Herceptin loaded and breast cancer targeted high molecular polymer ultrasound contrast agent and investigate their affinity for breast cancer in vitro. Methods The high molecular polymer PLGA-COOH ultrasound contrast agents were produced by the double emulsion technique. Herceptin was covalently linked to the PLGA-COOH nanoparticle surface using a carbodiimide technique. Its physical property was determined. The combination of Herceptin with the PLGA-COOH nanoparticle was proved by immunofluorescent assay,and the PLGA-COOH nanoparticle targeting specifity to breast cancer cells was observed with light microscope and confocal microscope,normal PLGA-COOH nanoparticle was served as control group. Results The diameter range of targeted high molecular polymer ultrasound contrast agents was 466.8～857.6 nm, the average diameter was (662.2 ± 69. 7) nm,85.9% of the diameter was in the range. Green punctiform fluorescence was observed by fluorescence microscope,and the conjugation of the targeted high molecular polymer ultrasound contrast agents with MCF-7 cells was tight while the conjugation in control group was negative. Conclusions The Herceptin loaded PLGA-COOH targeted ultrasound contrast agents was prepared successfully. The targeted ultrasound contrast agents can bind to breast cancer effectively in vitro.

Objective To explore the bioeffects of canine myocardium under microbubble destruction via diversity of ultrasonic intension, in order to optimize ultrasonic intension for experiments. Methods Nine mongrel dogs were randomly divided into 3 groups. Ultrasound (1 MHz) in diversity of different intension (0.5 W/cm~2, 1.0 W/cm~2, 2.0 W/cm~2) was applied to expose canine myocardium after intravenous injection microbubbles of 2.0 ml. All the dogs were killed after being exposed for 5 min. The myocardium was harvested for HE staining and observed with transmission electron microscope for the tissue microstructures. Results The myocardium of hyperemia, disfiguration and necrosis wer observed in all groups. Myocardial edema but not hemorrhage appeared with 0.5 W/cm~2 , mild myocardial hemorrhage and slight inflammatory cell infiltration happened with 1.0 W/cm~2, whereas obvious hemorrhage and certain degree of inflammatory cell infiltration occurred with intension of 2.0 W/cm~2. With the augmentation of ultrasonic intension, myocardium trend to aggravate. Conclusion Ultrasound of diversity intension can induce different bioeffects of canine myocardium. Ultrasound mediated microbubble destruction with the intensity of 1.0-2.0 W/cm~2 can provoke a certain degree of inflammatory reaction with mild myocardial damage.

Relapse after allogeneic hematopoietic stem cell transplantation (allo-HSCT) remains a main question on treatment failure.Current strategies for management that usually include salvage chemotherapy,donor lymphocytic infusion and second transplantation.Our study assessed the efficacy of decitabine (DAC) for treating patients with acute lymphoblastic leukemia (ALL) who relapsed after allogeneic hematopoietic stem cell transplantation (allo-HSCT).We retrospectively analyzed the outcomes of 12 patients with relapsed ALL after allo-HSCT who received DAC therapy.Nine patients received DAC combined with chemotherapy and donor stem cell infusion,and 3 patients received single-agent DAC.Ten of the 12 patients achieved complete remission (CR),1 achieved a partial remission (PR),and 1 had no response (NR) after treatment at the latest follow-up (LFU),the median survival was 11.2 months (range,3.8-34,7 months).The 1-and 2-year overall survival (OS) rates were 50％ (6/12) and 25％ (3/12),respectively.Five patients were still alive;4 had maintained CR and 1 was alive with disease.Patients with Philadelphia chromosome-positive ALL had higher survival rate than patients with Philadelphia chromosome-negative ALL (57.1％ vs.20％).No aggravated flares of graft-versus-host disease (GVHD) were observed during DAC treatment.Therefore,DAC may be a promising therapeutic agent for ALL recurrence after allo-HSCT.