A novel reverse emulsification-phase transition process was designed to prepare alginate-based nanocapsules in the present study. Ca alginate nanocapsules with an average hydrodynamic diameter below 300 nm were obtained by adjusting surfactant and alginate concentration. Instrumental analysis observations revealed a morphologically spherical shape with a relatively uniform size distribution without aggregation.?-potential measurements indicated an topically electrostatically negative nature of capsules in neutral aqueous solution. These nanocapsules were proven to have an equivalent potency to stimulate the maturation of dendritic cells originated from CD14 positive cells isolated from human peripheral blood. Protein loading via covalent conjugation was demonstrated feasible with EDC/NHS facilitated reaction between carboxyl and amino groups. These nanocapsules are considered to have important applications as a new adjuvant in cancer cell therapy, vaccine design and targeted drug delivery for active components of protein nature.

The combination of Glycyrrhizae Radix et Rhizoma and Aconiti Lateralis Radix Preparata can increase efficacy and decrease toxicity. This study started from the phenomena of protein self-assembly in the mixed decoction of Glycyrrhizae Radix et Rhizoma with Aconiti Lateralis Radix Preparata. The attenuated mechanism was explored between the combination of Glycyrrhizae Radix et Rhizoma and Aconiti Lateralis Radix Preparata by using the protein of Glycyrrhizae Radix et Rhizoma and aconitine which was the major toxic component of Aconiti Lateralis Radix Preparata. Glycyrrhizae Radix et Rhizoma protein with aconitine could form stable particles which particle mean diameter was (206.2 ± 2.02) nm and (238.20 ± 1.23) nm at pH 5.0 in normal temperature. Through the mouse acute toxicity experiment found that injection of aconitine monomer all mice were killed, and injection of Glycyrrhizae Radix et Rhizoma protein-aconitine particles with the same content of aconitine all mice survived. Survey the stability of Glycyrrhizae Radix et Rhizoma protein-aconitine shows that the colloid particles is stable at room temperature, and it has the possibility to candidate drug carrier. Glycyrrhizae Radix et Rhizoma protein can reduce the toxicity of aconitine through self-assembly.
Aconitum ; chemistry ; toxicity ; Animals ; Drugs, Chinese Herbal ; toxicity ; Female ; Glycyrrhiza ; chemistry ; toxicity ; Male ; Mice ; Mice, Inbred ICR ; Plant Proteins ; chemistry ; isolation & purification ; toxicity ; Rhizome ; chemistry ; toxicity

OBJECTIVETo investigate the molecular basis for a novel human leukocyte antigen (HLA) allele B*5827.

METHODSDNA from the proband was analyzed by polymerase chain reaction-sequence specific oligonucleotide (PCR-SSO) typing. The amplified product was sequenced bidirectionally.

RESULTSAbnormal HLA-B locus was observed and its nucleotide sequence was different from the known HLA-B allele sequences, with highest homology to HLA-B*5820 allele. It differs from HLA-B*5820 by 8 nucleotide substitutions in exon 3, i.e., nt 290 (G > C), nt 346 (T > A), nt 390 (A > C), nt 404 (G > C), nt 413 (C > G), nt 471 (A > G), nt 486 (A > G) and nt 487 (C > A), resulting in an amino acid change from ser > arg at nt 97, phe >tyr at nt 115, ser > arg at nt 130, thr > ala at nt 157 and thr > glu at nt 162. Nucleotide differences of nt 404 (G > C) and nt 413( C > G) did not change amino acid.

CONCLUSIONThe sequences of the novel allele have been submitted to GenBank (access No.GU071234). A novel HLA class I allele B*5827 has been officially assigned by the WHO HLA Nomenclature Committee in Jan. 2010.

Alleles ; Base Sequence ; Cloning, Molecular ; Genotype ; HLA-B Antigens ; chemistry ; genetics ; Humans ; Molecular Sequence Data ; Polymerase Chain Reaction ; Sequence Analysis, DNA

OBJECTIVETo explore the diagnosis, treatment, and prognosis of prostatic malignant mesenchymal tumors (PMMT).

METHODSWe retrospectively analyzed the clinical and follow-up data about 20 cases of PMMT and reviewed the literature relevant to the diagnosis, treatment, and prognosis of the disease.

RESULTSBased on the results of pathology and immunohistochemistry, the 20 PMMT cases included leiomyosarcoma (n = 7), rhabdomyosarcoma (n = 5), prostatic stromal sarcoma (n = 3), chondrosarcoma (n = 1), and undifferentiated PMMT (n = 4). Twelve of the patients were treated by radical prostatectomy (3 concurrently by sigmoid colostomy and 1 by cystostomy), 2 by pelvic tumor resection following arterial embolization, 1 by total pelvic exenteration, 1 by colostomy with pelvic lymph node biopsy, and 4 by conservative therapy because of metastasis to the lung, pelvis and bone. Of the 20 patients, 9 died of systemic metastasis within 3 months after treatment, 3 died at 6, 7, and 14 months, respectively, 3 survived with tumor for 5, 11, and 12 months, respectively, 2 survived without tumor for 12 and 24 months so far, all subjected to periodic chemotherapy postoperatively, and 3 lost to follow-up.

CONCLUSIONPMMT is a tumor of high malignancy and rapid progression, for which transrectal ultrasound-guided biopsy remains the main diagnostic method. The clinical stage of the tumor is an important factor influencing its prognosis and the survival rate of the patients can be improved by early diagnosis and combined therapy dominated by radical prostatectomy.

Combined Modality Therapy ; methods ; Humans ; Immunohistochemistry ; Male ; Mesenchymoma ; mortality ; pathology ; therapy ; Prognosis ; Prostatectomy ; Prostatic Neoplasms ; mortality ; pathology ; therapy ; Retrospective Studies

OBJECTIVENimotuzumab is a humanized monoclonal antibody targeted against epidermal growth factor receptor (EGFR). Recent clinical studies show that patients with malignant gliomas could benefit from nimotuzumab treatment. The aim of the present study was to evaluate the efficacy and side effects of nimotuzumab in combination with chemotherapy for patients with malignant gliomas.

METHODSThe patients received 200 mg of nimotuzumab infusion intravenously over 60 minutes once weekly for the first eight weeks and then once every two weeks until unacceptable toxicity or tumor progression occurred. Individualized chemotherapy was administered based on O(6)-methylguanine-DNA methyltransferase (MGMT) expression and previous chemotherapy responses in combined with nimotuzumab.

RESULTSFourteen patients received a total of 122 times of nimotuzumab ranging from 2 to 20 (median 7.5 times). Combined chemotherapy regimens included: continuous 21-day temozolomide (10 cases), standard 5-day temozolomide (2 cases), teniposide plus cisplatin (1 case), and teniposide plus nimustine (1 case). Partial response (PR) and stable disease (SD) were found in 3 patients (21.4%)and 6 patients (42.9%), respectively. Disease control rate (PR + SD) was 64.3%. The median progression-free survival (PFS) was 4 months (95%CI: 0.7 - 7.3) and PFS at 6 months was 30.6%. The most common toxicities include grade I-II neutropenia (2 cases), thrombocytopenia (2 cases), lymphopenia (1 case), nausea and vomitting (3 case) and asymptomatic transaminase increase (1 case). One patient developed grade IV neutropenia and thrombocytopenia. One patient developed nimotuzumab-related acneiform rash.

CONCLUSIONSNimotuzumab in combination with chemotherapy has moderate activity in patients with malignant gliomas and the toxicities are well tolerable, therefore, worth further investigation.

Adolescent ; Adult ; Antibodies, Monoclonal, Humanized ; administration & dosage ; adverse effects ; therapeutic use ; Antineoplastic Agents, Alkylating ; adverse effects ; therapeutic use ; Antineoplastic Combined Chemotherapy Protocols ; adverse effects ; therapeutic use ; Astrocytoma ; drug therapy ; Child ; Cisplatin ; administration & dosage ; adverse effects ; Dacarbazine ; adverse effects ; analogs & derivatives ; therapeutic use ; Disease-Free Survival ; Female ; Glioblastoma ; drug therapy ; Glioma ; drug therapy ; Humans ; Infusions, Intravenous ; Male ; Nausea ; chemically induced ; Neutropenia ; chemically induced ; Nimustine ; administration & dosage ; adverse effects ; Teniposide ; administration & dosage ; adverse effects ; Thrombocytopenia ; chemically induced ; Young Adult

Aim To observe the inhibitory effect of Alpha-momorcharin (α-MMC) on the inflammatory cytokine storm of Ml-type inflammatory macrophages induced by LPS and explore its possible targeting mechanism. Methods Western blot was used to detect the expression of WIL2-S B lymphocytes, H9 T lymphocytes, THP-1 monocytes and M0 macrophages LRP1 receptor protein. CCK-8 method was used to detect the survival rate of the four cells. ELISA was used to detect the expression level of inflammatory cytokines in Ml macrophages. Western blot was used to detect the expression of TLR4 signaling pathway-related protein in Ml macrophages. Results Macrophages had a high density of LRP1 receptors consistent with monocytes; the survival rate of α-MMC on the four cells was positively correlated with the density of this receptor; α-MMC inhibited the expression of inflammatory cytokinesTNF-α, IL-lβ, IL-6, IL-8, MlP-lα and MCP-1 in Ml macrophages in a dose-and time-dependent manner; α-MMC showed significant inhibition to TAKl/pTAK1, p-JNK, p-APl and p-p65 signaling proteins of the TLR4 signaling pathway, and this inhibition could be blocked by the LRP1 receptor blocker RAP. Conclusions α-MMC selectively inhibits macrophage inflammatory cytokine synthesis by inhibiting TAK1 of the TLR4 signaling pathway, which in turn inhibits the downstream NF-ΚB and MAPK pathways, mediated by the LRP1 receptor. The selective immunosuppressive effect of α-MMC on macrophages may make it a very promising agent for the treatment of acute infectious macrophage activation syndrome (MAS).

OBJECTIVETo explore the prevalence and the clinical characteristics of mitochondrial gene mutation A3243G (mt tRNA(Leu(UUR)) 3243 A-->G) in patients with type 2 diabetes mellitus (DM2) in China.

METHODSFour hundred and twenty-eight cases of DM2 patients were selected randomly. One hundred and eighty-eight individuals were healthy controls. The mutation was assayed by PCR-restriction fragment length polymorphism technique. The target fragments of PCR were digested with restriction endonuclease Apa I.

RESULTSmt tRNA(Leu(UUR)) 3243A-->G gene mutation was found in 2 of 428 patients with DM2, but not found in the controls. Further investigation of the relatives of the 2 patients' families revealed that 3 members were the carriers of mt tRNA A3243G gene mutation and the patients with diabetes. In addition, one proband and her son were characterized with the syndrome of mitochondrial encephalomyopathy with lactic acidosis. The diabetes of these patients is frequently accompanied by hearing impairment or deafness with maternal inheritance.

CONCLUSIONThe prevalence of the mitochondrial gene A3243G mutation is 0.47% in DM2 patients in China. The data acquired in this study suggest that the clinical phenotype of these patients with A3243G should be heterogeneous.

Adult ; Aged ; DNA, Mitochondrial ; genetics ; Diabetes Mellitus, Type 2 ; genetics ; Female ; Humans ; Male ; Middle Aged ; Mutation ; RNA, Transfer, Amino Acyl ; genetics

OBJECTIVETo develop a system to rescue virus by intracellular expression of T7 RNA Polymerase.

METHODSThe gene of T7 RNA Polymerase was amplified and cloned to VR1012 by molecular biological technology. The expression plasmid VR-1a was then identified. VR-1a and EV71 infectious plasmid were co-transfected in Vero cell. CPE was observed and viral gene viral antigen were detected.

RESULTSThe gene of T7 RNA Polymerase was successfully cloned into vector VR1012. Vero cell developed to CPE after being transfected VR-1a and EV71 infectious plasmid. EV71 gene was amplified by RT-PCR from the culture. EV71 antigen was also detected by ELISA.

CONCLUSIONThe method can be used to rescue virus. It could apply to immunologic research of EV71 DNA vaccine.

Animals ; Cercopithecus aethiops ; DNA-Directed RNA Polymerases ; genetics ; metabolism ; Enterovirus A, Human ; genetics ; physiology ; Gene Expression ; Genetic Engineering ; methods ; Genetic Vectors ; genetics ; metabolism ; HeLa Cells ; Humans ; Plasmids ; genetics ; metabolism ; Transfection ; Vero Cells ; Viral Proteins ; genetics ; metabolism ; Virus Replication

OBJECTIVETo develop attenuated Salmonella which harboring enterovirus 71 (EV71) VP1 gene.

METHODSThe plasmid which expressed VP1 protein of EV71 was constructed by gene recombination. Cellular expression was assessed by Western Blot analysis. The recombinant plasmid was then transformed into attenuated Salmonella SL7207.

RESULTSEV71 VP1 gene sequence was inserted into a eukaryotic expression plasmid VR1012. VP1 protein was detected by Western Blot analysis in the culture supernatant. And the attenuated Salmonella harbored the plasmid stable.

CONCLUSIONThe plasmid was constructed successfully and it can express effectively in vitro. The bacteria which harboring the plasmid were constructed successfully. This has provided a basis for further research of an oral EV71 vaccine.

Capsid Proteins ; genetics ; metabolism ; Enterovirus A, Human ; genetics ; Gene Expression ; Genetic Engineering ; Genetic Vectors ; genetics ; metabolism ; Salmonella ; genetics ; metabolism

BACKGROUNDFew reports have evaluated the efficacy of re-operation for relapse after initial surgery for hepatocellular carcinoma (HCC) with bile duct thrombosis (BDT). The aim of this study was to investigate the efficacy of initial surgery and subsequent re-operation for HCC with BDT, and their effects on prognosis.

METHODSThe clinical data of 880 patients with HCC, including 28 patients with BDT, who underwent radical hepatectomy between 1998 and 2008 in our hospital, were reviewed. The effects of BDT and re-operation on prognosis were retrospectively analyzed.

RESULTSThe 1-, 3- and 5-year survival rates were 89.3%, 46.4% and 21.4%, respectively, in 28 patients with BDT versus 91.4%, 52.9% and 20.9% in 852 patients without BDT (P>0.05). Six patients with BDT underwent re-operation after disease relapse, and their survival time was significantly longer than those who did not undergo re-operation (P<0.05). Multivariate analysis indicated that portal vein invasion and tumor size were independently associated with tumor relapse and prognosis (P<0.05). Univariate analysis and multivariate analyses showed that obstructive jaundice was not significantly correlated with tumor relapse or prognosis (P>0.05).

CONCLUSIONSHepatectomy plus BDT removal is an effective treatment option for HCC with BDT. Obstructive jaundice is not a contraindication for surgery. Re-operation after relapse can provide good outcomes if the cases are appropriately selected.

Adult ; Bile Ducts ; pathology ; Carcinoma, Hepatocellular ; diagnostic imaging ; surgery ; Female ; Hepatectomy ; Humans ; Liver Neoplasms ; diagnostic imaging ; surgery ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Radiography ; Thrombosis ; surgery ; Treatment Outcome ; Ultrasonography