OBJECTIVEIn this study, the authors investigated inhibition of coxsackievirus B (CVB) gene expression using antisense oligonucleotides complementary to the 5' NCR, translation initiation codon and structural protein coding sequences and also observed the dose-response of the sequence specific inhibition of CVB plaque formation by antisense oligonucleotides.

METHODSAntiviral activities of these oligonucleotides were evaluated by using plaque reduction assay, yield reduction assay, cytopathic effect (CPE) and Western blot analysis. The cells were treated with random oligonucleotides as a specificity control.

RESULTSAt a screening concentration of 5 micromole, 6 of the phosphorothioate oligonucleotide demonstrated some reduction of virus replication relative to untreated cells. 70%-90% inhibition of virus at 0.1 MOI (multiplicity of infection), 50% inhibition of virus infection at 10 MOI. The levels of the VP1 were reduced in CVB-infected cells treated with Scb561 and Scb733. VP1 was significantly reduced after treatment with 0.625 micromole Scb561 and almost undetectable in cells treated with 2.5 micromole Scb561. Dose response experiments implied that sequence specific oligonucleotide doses were related to effect on inhibition of CVB3 infection. When oligonucleotide doses were increased from 1.25 to 5 micromole, 75% to 90% inhibition were observed with Scb561 and 65% to 80% inhibition with Scb733, whereas random control failed to inhibit CVB replication (8% inhibition for each). CONCLUSION The present studies showed that antisense oligonucleotides against internal ribosome entry site (IRES) and translation initiation codon were capable of specifically inhibiting the synthesis of viral protein and subsequent productive CVB replication.The selective inhibition using antisense oligonucleotide might lead to development of an effective antiviral agent for future clinical evaluation.

5' Untranslated Regions ; genetics ; Dose-Response Relationship, Drug ; Enterovirus B, Human ; drug effects ; genetics ; Gene Expression ; drug effects ; HeLa Cells ; Humans ; Oligonucleotides, Antisense ; pharmacology ; Ribosomes ; metabolism ; Viral Structural Proteins ; biosynthesis ; genetics ; Virus Replication ; drug effects

Abnormalities, Multiple ; surgery ; Adolescent ; Humans ; Male ; Spleen ; abnormalities ; Testis ; abnormalities

ObjectiveSLC2A9 is a novel identified urate transporter that affects serum uric acid levels. The present study is aimed to investigate rs7442295 polymorphism in intron 6 of SLC2A9 in a population of Chinese male gout or hypemricaemia subjects. MethodsA total of 268 gout patients and 288 healthy male volunteers were included. Blood pressure, body mass index(BMI), serum uric acid, glucose, lipid,urea and creatine were detected. DNA was purified from peripheral blood and the rs7442295 polymorphism was evaluated using high resolution melting ( HRM ) analysis and direct sequencing. Data were analyzed with t test or chi-square test. Results A/A and A/G genotypes were unambiguously distinguished with HRM technology. The occurrence of the homozygous type (G/G) was completely absent among the study population.The prevalence of the A/A and A/G genotype was 96.2％ and 3.8％ respectively. However, no significant differences of genotype frequencies were found in gout patients and normal subjects(x2=0.003, P=0.82; x2=0.003, P=1.00). But the serum uric acid levels in individuals with the A/G genotype[(293±100) μmol/L]were significantly lower than those with the A/A genotype[(392±133) μmol/L](t=2.426, P＜0.01 ). The A/G genotype frequency was significantly higher in the low-uric acid group than in the high uric-acid group (x2=6.279, P=0.01 ). Genotyping based on HRM was fully concordant with sequencing. Conclusion The polymorphism rs7442295 in SLC2A9 may be a genetic marker to assess risk of hyperuricemia among Chinese male Hart population. HRM is a simple, fast, reliable and close-tube technology for genotyping.

Objective To observe the expression levels of matrix metalloproteinase-2 and 9 (MMP-2, MMP-9) in wound tissue and wound effusion in patients with chronic pressure ulcers treated with an alternating pres- sure relief mattress,and to correlate these observations with wound healing.Methods A total of 24 patients with chronic pressure ulcers were recruited and divided into two groups :one treated with an alternating pressure relief mat- tress and the other without,in addition to conventional treatment for pressure ulcers.The expression of MMP-2 and MMP-9 in the wound tissue and its exudate were amplified with a reverse transeriptase-polymerase chain reaction (RT-PCR) and analyzed using quantitative gelatin zymography.Wound healing was also observed and assessed using the pressure ulcer scale for healing (PUSH).Results The wounds were observed to heal well in both groups,as indicated by a significant decrease in the PUSH scores.Statistical analysis revealed a significant difference between the two groups with regard to the PUSH score on the 21st day after initiation of the study.With the healing of the pressure ulcers,the expression both MMP-2 and MMP-9 decreased.On the seventh and the 21st days,the expression of MMP-2 and MMP-9 in patients treated with an alternating pressure relief mattress was significantly less than in those without the mattress.Conclusion MMP-2 and MMP-9 can be biomarkers for the healing of pressure ulcers. An alternating pressure relief mattress is helpful for treating pressure ulcers.

BACKGROUND: Although the porcine small intestinal submucosa is very similar to the skin in the structure, its pore size and porosity are not beneficial to the growth of seed cells as the skin does. OBJECTIVE: To observe the morphological and cytocompatibility of porcine small intestinal submucosa after chemical modification using 1-ethyl-3-(3-dimethylaminopropyl) carbod imide (EDC). METHODS: A porcine small intestinal submucosa sample was immersed in 3% acetic acid solution containing 0.2% pepsin to make 1%, 2%, 3%, 4% small intestine submucosa solutions. After magnetic stirring and freeze-drying, small intestinal submucosa sponge was obtained and modified by cross-linking with 50, 100, 150 mmol/L EDC. Based on the detection of pore size and water absorption, we selected the best concentrations of small intestinal submucosa and EDC, which were further used for cell culture. Passage 2 bone marrow mesenchymal stem cells were cocultured with the small intestinal submucosa sponge, and observed under scanning electron microscope at 1, 2, 3 weeks after co-culture. RESULTS AND CONCLUSION: When cross-linked with 100 mmol/L EDC, the small intestinal submucosa sponge at a mass concentration of 1% showed a reasonable structure and good elasticity with no appearance of voids, and the pore size ranged 100-150 μm. Moreover, the small intestinal submucosa sponge at a mass concentration of 1% showed a 0.35-fold increase in the compared with that at a mass concentration of 2%, and its structure was more conducive to water flows and changes. The small intestinal submucosa sponges at a mass concentration of 3% and 4% showed no difference in the water absorbing capacity. After cross-linked with 100 mmol/L EDC, the small intestinal submucosa sponge at a mass concentration of 1% showed the best structure, pore size and water absorption, which were used for cell culture. At 3 weeks after cell culture, cell deformation was relatively intact and fast; there were many cells in the pores that were relatively large and approximately spindle-shaped, while there were less cells around the pores that were relatively small and disk-shaped. A paving stone-like alteration was observed in cells that covered the most of the scaffold surface with a large number of particle-like substances, especially in the site of cell concentration. All these findings indicate that EDC-modified small intestinal submucosa sponge has good cytocompatibility.

OBJECTIVETo evaluate the effectiveness of chemotherapy (CT) and autologous hematopoietic stem cell transplantation (ASCT) as post-remission treatment for adult acute lymphoblastic leukemia (AL) patients.

METHODSSeventy-four ALL patients achieved first complete remission (CR(1)) with induction therapy, and then received early-stage sequential intensive consolidation chemotherapy. After that, 40 patients received chemotherapy (CT group) and 34 received ASCT (ASCT group) as post-remission treatment. The median follow-up was 20.5 months. The rates of leukemia free survival (LFS), overall survival (OS) and relapse were compared between the two groups.

RESULTS(1) The median LFS and OS were 14.0 and 20.6 months respectively for CT group and both were more than 53.5 months for ASCT groups. (2) Relapse occurred in 28 patients (70%) in CT group in a median time of 8.5 months (range, 1-72 months) and 20 of them (71.43%) relapsed within 1 year. Eleven patients (32.35%) relapsed in ASCT group, in a median time of 6 (2-30) months after transplantation. (3) There was no statistic difference in LFS, OS and relapse rate at 1 year between CT and ASCT groups (P > 0.05), whereas both LFS and OS at 3 and 5 years for ASCT group were significantly better than those for CT group (P < 0.05). Relapse rate for ASCT group was lower than that for CT group. (4) Higher LFS and OS and lower relapse rate were found for those who received monoclonal antibody purged autografts followed by immunotherapy and (or) maintenance therapy after ASCT (P < 0.05).

CONCLUSIONSEarly sequential intensive consolidation chemotherapy followed by auto-HSCT could significantly reduce late relapse rate for adult ALL patients, and those received ex vivo purged autografts and immunotherapy and (or) maintenance therapy after ASCT have lower late relapse rate and superior survival.

Adolescent ; Adult ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Female ; Follow-Up Studies ; Hematopoietic Stem Cell Transplantation ; Humans ; Male ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; drug therapy ; surgery ; Retrospective Studies ; Transplantation, Autologous ; Treatment Outcome

OBJECTIVETo evaluate the treatment outcome of HLA-identical sibling allogeneic hematopoietic stem cell transplantation (allo-HSCT) for chronic myelogenous leukemia (CML) patients in first chronic phase (CP(1)).

METHODSFifty-one patients with CML-CP(1) received HLA-identical sibling allo-HSCT with conditioning regimens of TBI plus Cy or Bu plus Cy. Allogeneic peripheral blood stem cell transplantation (PBSCT) and bone marrow transplantation (BMT) were performed for 28 and 23 patients, respectively. The median follow-up duration was 1434 (60 - 4062) days.

RESULTSFifty (98.0%) patients were successfully engrafted. Transplant-related mortality occurred in 8 (15.7%) patients. Acute graft-versus-host disease (aGVHD) occurred in 35 (68.6%) patients and 11 (21.6%) patients were grade II-IV, while chronic GVHD (cGVHD) did in 17 (37.8%) patients. Five (7.4%) patients relapsed. The 5-year probability of disease-free survival (DFS) was (79.2 +/- 6.4)%. There was no significant difference in 5-year DFS, death rate and treatment related syndromes between the two conditioning regimens (P > 0.05), and in 5-year DFS, relapse rate and death rate between two transplant choices (P > 0.05). However, the rate of relapse was lower in Bu/Cy group (P < 0.01) and the rate of cGVHD was higher in allo-PBSCT group (P < 0.05).

CONCLUSIONSAllo-HSCT can cure a significant proportion of patients with CML-CP(1). There was no significant difference in DFS between the two different conditioning regimens and between the different transplant choices. Donor lymphocyte infusion is a therapeutic alternative for CML patients relapsed after transplantation.

Adult ; Female ; HLA Antigens ; Hematopoietic Stem Cell Transplantation ; methods ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; surgery ; Male ; Middle Aged ; Recurrence ; Siblings ; Transplantation Conditioning ; Transplantation, Homologous ; Treatment Outcome

OBJECTIVETo investigate the genetic diversity in Chinese populations. And HLA-A, -B alleles and haplotypes of 190 unrelated healthy individuals of Tu nationality from Wufeng county Hubei province were identified for the associated studies of HLA gene polymorphism and disease.

METHODSThe high-resolution typing methods--sequence-based typing(SBT) was used to define the most polymorphism of exons 2 and 3 of the HLA-A, -B locus alleles. The allele and haplotype frequencies were calculated by maximum likelihood estimation with Arlequin software.

RESULTSHLA-A, -B alleles were found to be in Hardy-Weinberg equilibrium(P>0.05). A total of 26 HLA-A and 41 HLA-B alleles were detected. The most frequent alleles were A*0201(0.16053), A*110101(0.14737), A*24020101(0.14211), B*4001(0.14737), B*4601(0.13947), followed by A*0207(0.08947), A*0206(0.08158), B*1301(0.07632), B*5801(0.08947), B*1501(0.09737). The frequencies of following alleles to be A*330301(0.05526), B*1502(0.05526), B*3501(0.05263) were all higher than 0.05. The extensive HLA-A-B haplotypes were observed, and the most common haplotypes were A*0202-B*4001(0.04196), A*0201-B*4601(0.03625).

CONCLUSIONIn the present study, we first analyzed the HLA-A, B gene typing with SBT, all of these results will be the basic and reference data for Tu race, and also will have the applications available to trace the population migration, clinical organ transplantation, disease-associated study, HLA genetic feature and forensic identification.

Alleles ; Asian Continental Ancestry Group ; ethnology ; genetics ; China ; ethnology ; Ethnic Groups ; genetics ; HLA Antigens ; analysis ; genetics ; HLA-A Antigens ; analysis ; genetics ; HLA-B Antigens ; genetics ; Humans

OBJECTIVETesticular mixed nonseminomatous germ cell cancer (TMNGCC) is rarely reported. This study aimed to explore the clinical symptoms, pathological characteristics and treatment methods of TMNGCC.

METHODSWe analyzed the clinical data of 1 case of TMNGCC, observed its pathological characteristics under the light microscope by histology, cytochemistry, immunohistochemistry and immune marking, and investigated the clinical features of such tumors by reviewing the relevant literature.

RESULTSThe patient presented with a chief complaint of painless testicular swelling for 3 years. Histopathological examinations revealed a tumor of papillary, fissural or adenoid structure, with large polygonal or columnar cells with one or more irregular vesicular nuclei, the nuclear membrane clear, the cytoplasm eosinophilic or basophilic, and the interstitium infiltrated by a few lymphocytes. Here are the immunohistochemical results: CD117 -, CK8-18 + +, CD30 + +, CK + + +, vimentin -, PLAP +/-, P53 +, AFP + and EMA + +. The tumor was pathologically diagnosed as teratogenic embryonic testicular cancer, and treated by radical surgery, followed by adjuvant chemotherapy according to the treatment of TMNGCC. One-year follow-up found the patient to be alive.

CONCLUSIONTMNGCC is a rare malignant tumor, mostly with unobvious clinical symptoms. Its diagnosis primarily depends on physical examination, ultrasonography, CT, and measurement of serum tumor markers; its confirmation necessitates pathological examination, and its first-choice treatment is surgical resection.

Adult ; Humans ; Male ; Neoplasm Staging ; Seminoma ; pathology ; Testicular Neoplasms ; pathology

OBJECTIVETo establish a new culture system for mouse tooth germs in chick embryo.

METHODSThe mandibular first molar germ fragments of 15 embryonic days' Kunming mouse embryo were implanted into the lateral mesenchyme of 4-5 days' chick embryo wing buds in ove. Eggs were reincubated and implanted tissues were examined by histochemistry.

RESULTSThe cultured tooth germ development continued from cap stage to latest bell stage. The ameloblast and the odontoblast all differentiated maturely and secreted matrix.

CONCLUSION4-5 days' wing buds chick embryo could serve as developing the mouse tooth germs and demonstrate well physiological process of differentiation and morphogenesis.

Ameloblasts ; Animals ; Chick Embryo ; Mice ; Molar ; embryology ; Odontoblasts ; Tooth Germ ; embryology