Effectiveness of HM-3-HSA on Inhibiting Cancer Cell Migration and Metastasis.

BACKGROUND: Metastasis is the major cause of treatment failure in cancer patients and cancer- associated death, and an antimetastatic drug would be a beneficial therapy for cancer patients. HM-3-HSA is a fusion protein which improved the pharmacokinetics of HM-3 and exerted antitumor and anti-angiogenesis activity in multiple tumor models. However, the efficacy of HM-3-HSA in cancer cell migration and metastasis has not been elucidated. MATERIALS AND METHODS: Herein, high-cell density fermentation of Pichiapink strain expressing HM- 3-HSA was performed for the first time. Then, the desired protein was purified by Butyl Sepharose High performance, Capto Blue, Phenyl Sepharose 6FF HS and DEAE Sepharose FF. Furthermore, the effect of HM-3-HSA on the migration and invasion of cancer cells was also evaluated, and B16F10 metastasis model was established to detected the anti- metastasis effect of HM-3-HSA in vivo. RESULTS: The results indicated that the yield of HM-3-HSA was 320 mg/L in a 10 L fermenter, which was a 46% increase over that expressed in flask cultivation. The desired protein was purified by fourstep, which yielded a 40% recovery of a product that had over 99% purity. Purified HM-3-HSA significantly suppressed the migration and invasion of HCT-116, SMMC-7721 and B16F10 cell lines. CONCLUSION: On the other hand, in the B16F10 metastasis model, HM-3-HSA significantly inhibited pulmonary metastases of B16F10 cells, suggesting that HM-3-HSA exerted the anti-metastasis effect in vivo.

HM-3-HSA exhibits potent anti-angiogenesis and antitumor activity in hepatocellular carcinoma.

HM-3-HSA is an antitumor fusion protein which improved the pharmacokinetics of HM-3. Previous studies reported that HM-3-HSA enhanced antitumor activity of HM-3 in melanoma cells. However, the efficacy and the mechanism of HM-3-HSA in hepatocellular carcinoma, especially its effect on tumor angiogenesis, have not been elucidated. Herein, we showed that HM-3-HSA significantly inhibited the H22 and SMMC-7721 tumor xenografts growth and tumor angiogenesis in vivo, indicating the antitumor activity exerted by HM-3-HSA was closely corrected with its potency on tumor angiogenesis. To investigate the anti-angiogenic mechanism, we evaluated the efficacy of HM-3-HSA in HUVECs in vitro. The results showed that multiple steps of tumor angiogenesis, including endothelial cell proliferation, migration, invasion and tube formation, were substantially inhibited by HM-3-HSA. Mechanism investigations revealed that HM-3-HSA could bind HUVECs via integrin αvß3 and α5ß1 and inhibited phosphorylation of the downstream protein kinases including FAK, Src and PI3 K. Our study was the first to report the activity of HM-3-HSA against hepatocellular carcinoma and tumor angiogenesis as well as the underlying mechanism by which HM-3-HSA to exert its anti-angiogenic activity.

The Effectiveness of Preoperative Preparation for Improving Perioperative Outcomes in Children and Caregivers.

Most children experience significant anxiety during the preoperative period. Greater preoperative anxiety may be related to a higher incidence of negative behaviors. This study aimed to develop a family-centered preoperative preparation program and to evaluate the effects of this program on children's preoperative emotional behaviors, postoperative behavior, and posthospital behavior, and on caregiver anxiety. A prospective, randomized controlled study was conducted. The population consisted of children who underwent minor surgery and their caregivers. The control group received standard care, and the experimental group received standard care plus preoperative preparation, which included a tour, a cartoon video depicting a boy's surgical journey, and familiarization with medical equipment. Children's emotional behaviors and caregiver anxiety were measured at the preoperative visit, in the preoperative holding area, and at induction of anesthesia. Postoperative behavior was measured when children were in the recovery room, and the researcher also contacted caregivers 2 weeks after the surgery to assess the children's behavior at home. A linear mixed-effects model results showed that as the surgery approached, the experimental group had fewer and more stable preoperative emotional behaviors (least squares means of preoperative emotional behaviors from preoperative visit to induction of anesthesia = 10.01-10.95). However, the control group exhibited significantly increased preoperative emotional behaviors as the surgery approached (least squares means of preoperative emotional behaviors from the preoperative visit to induction of anesthesia = 7.87-12.23). Family-centered preoperative preparation can effectively improve children's negative emotional behaviors from their time in the preoperative holding area to the induction of anesthesia.

Mapping of Taenia solium TSOL18 antigenic epitopes by phage display library.

Taenia solium is a cestode parasite that causes cysticercosis in humans and pigs. TSOL18 has been identified as a host-protective oncosphere antigen. To obtain mouse monoclonal antibodies (mAbs) against TSOL18 and to map its antigenic epitopes are potentials to develop a vaccine for the prevention of T. solium infection. In this study, mAbs were produced by the hybridoma technique using purified glycosylated TSOL18 produced in Pichia pastoris as the immunogen. mAb was used to define the B-cell epitopes of TSOL18 with phage-displayed random dodecapeptide library (Ph.D.-12), and some of the positive phage clones were sequenced and analyzed. The predominant mimotopes were ETTKLQRFQAML (L1) found in 83%, followed by DHTXF in 15% (L2: DHTLFAASHNHR, DHTLFSTGHSHG, and DHTFMQRYHTHQ). Comparison of the peptide sequences with native TSOL18 protein sequence using Clustal W software showed that they did not completely match, suggesting that the ETTKLQRFQAML and DHTXF sequences should be conformational epitopes. The sera of mice immunized with the selected phage clones obviously recognized the TSOL18 protein. Meanwhile, sera collected from TSOL18-vaccinated pigs reacted to both epitopes in enzyme-linked-immunosorbent serologic assay test. Our work demonstrated that the antigenic epitope could be mapped through screening the phage-displayed peptide libraries with mAbs and a mimotope of TSOL18, which could provide an alternative approach for the diagnosis and development of a vaccine for T. solium.

[Observation on the ultrastructure of Taenia solium oncosphere].

OBJECTIVE: To observe the ultrastructure of Taenia solium oncospheres. METHODS: Patients infected with Taenia solium were de-wormed by decoction arecae and pumpkin seeds to get the mature proglottids and collect eggs. The eggs were treated with sodium hypochlorite to break the eggshells. Oncospheres were collected in Percoll (isoosmotic solution), and activated with artificial intestinal juice. The specimens were prepared with hot agar centrifugation for ultra-thin sections and observed with transmission electron microscopy (TEM) . RESULTS: T. solium oncosphere was in oval shape with a size of (14-17) microm x (10-13) microm. There were some irregular ecphymata or plicae on its surface. The hooks were composed of outer pellet layer, the middle fibrous layer and the central core marrow. Myoblasts, hook-forming cells and cerebral cortex cells were observed in the mature oncospheres. CONCLUSION: The ultrastructure of Taenia solium oncosphere is similar to that of Hymenolepis diminuta, with difference in hooks. There are binucleate cells which play a role in forming epithelium in the development of oncospheres.

[Adjuvant effect of CpG DNA recombinant plasmid on antigen of Cysticercus cellulosae].

In order to prove the adjuvant effect of CpG DNA recombinant plasmid, the total antibodies and their IgG2a subtype induced by antigen of Cysticercus cellulosae, and content of IL-4 and IFN-gamma secreted from splenic cell of mouse immunized were measured. The recombinant plasmids showed an adjuvant effect, and CpG2 was the best adjuvant among the plasmids. It is proved that the CpG DNA possesses a synergistic effect with AI(OH)3 and 206 adjuvant, and is an effective Th1 type adjuvant in mice.