This study was aimed to establish a quality evaluation method, quantitative analysis of multi-components with a single-marker (QAMS) to determine the contents of five saponins in Yao-Bi-Tong (YBT) capsules. Ginseno-side Rg1 was used as the internal standard; the relative correction factor (RCF) of notoginsenoside R1, ginsenoside Re, ginsenoside Rb1 and ginsenoside Rd were calculated and evaluated. The contents of 5 saponins were determined by the external standard method and QAMS, respectively. Rationality, feasibility and repeatability of the QAMS method were verified by comparing the results obtained from two different methods. The results showed that RCFs of notoginsenoside R1, ginsenoside Re and ginsenoside Rb1 to ginsenoside Rg1 against YBT capsules were 0.999, 1.228, 0.990 and 1.094, respectively, indicating good reproducibility. These results of two methods had no significant dif-ference. It was concluded that the QAMS method can be accurately, rapidly and reasonably used as a new quality assessment model for ginsenosides in YBT capsules.

BACKGROUNDThe use of a free, vascularized fibular graft is an important technique for the reconstruction of large defects in long bones. The technique has many advantages in strong, tubular bones; a more reliable vascular anatomy with a large vascular diameter and long pedicle is used, minimizing donor-site morbidity. Due to limitations in both fibular anatomy and mechanics, they cannot effectively be used to treat large limb bone defects due to their volume and strength.

METHODSFrom 1990 to 2001, 16 clinical cases of large bone defects were treated using vascularized double-barrel fibular grafts. Patients were evaluated for an average of 10 months after surgery.

RESULTSAll the patients achieved bony union; the average bone union took 10 months post surgery, and no stress fractures occurred. Compared with single fibular grafts, the vascularized double-barrel fibular grafts greatly facilitate bony union and are associated with fewer complications, suggesting that the vascularized double-barrel fibular graft is a valuable procedure for the correction of large bone defects in large, long bones in addition to enhancing bone intensity.

CONCLUSIONSThe vascularized double-barrel fibular graft is superior to the single fibular graft in stimulating osteogenous activity and biological mechanics for the correction of very large bone defects in large, long bones. Free vascularized folded double-barrel fibular grafts can not only fill up large bone defects, but also improve the intensity margin. Therefore, this study also widens its application and enlarges the treatment targets. However, in the case of bone deformability, special attention should be paid to bone fixation and protection of donor and recipient sites.

Adolescent ; Adult ; Bone Diseases ; pathology ; surgery ; Bone Transplantation ; methods ; Female ; Fibula ; pathology ; surgery ; Humans ; Lower Extremity ; pathology ; surgery ; Male ; Middle Aged ; Models, Biological ; Reconstructive Surgical Procedures ; methods ; Reproducibility of Results

Objective with the advantages of rapidity in detection protein, We selected the gender-specific amino acid sequence based on human SMCY and SMCX, cloned and expressed SMCY gender-specific fusion antigens. The rabbits were immunized with purified antigens to obtain the polyclonal antibodies. A new method was established for rapidly sex identification of forensic evidence samples by detecting SMCY antigens with the corresponding polyclonal antibodies. Methods We found three differential fragments by analyzing the sequence of human SMCY and SMCX. Then we cloned this three fragments and ligated as a new recombinant.This SMCY gender-specific fusion antigen gene was sub-cloned into pET-28a and expressed in Escherichia coli. The fusion antigen was purified by Ni-NTA column. The rabbits were immunized with purified antigen to produce the specific polyclonal antibodies.The reactivity of the polyclonal antibody was evaluated by ELISA and Western blotting. We developed a colloidal gold test strip for detecting the gender of human samples. Results We successfully selected gender-specific amino acid sequence, the SMCY gender-specific fusion antigen was expressed by prokaryotic expression and the polyclonal antibody was prepared by immunizing rabbit. The results of colloidal gold strip tests showed that there is a significant difference between male and female serums. Conclusion The results showed that the SMCY gender-specific fusion antigen could be recognized by the polyclonal antibody.The colloidal gold strip tests made by SMCY gender-specific fusion antigens and the corresponding polyclonal antibodies could be used for rapidly determining the gender of forensic evidence samples.

Objective To construct an expression plasmid of human papillomavirus type 11 E7 (HPV11-E7)/hurnan IFN?-2b fusion gene, to express the fusion gene in E.coli BL21, and pave way for further immunological study. Methods The recombinant plasmid was introduced into E.coli BL21, then the expression product was analyzed by SDS-PAGE and Western blotting after induction with isopropy-?-D-thiogalactoside (IPTG). Results The fusion gene of HPV11-E7 and human IFN?-2b was successfully cloned into pET-32a by a linker with the same sequence as we expected. The expressed fusion protein was confirmed by SDS-PAGE and Western blotting. Conclusions The successful construction of prokaryotic expression plasmid and expression of HPV11-E7/human IFN?-2b fusion gene enable further immunological study.

Objective To evaluate the clinical features and the strategy for the diagnosis and treat- ment of bilateral testicular tumor.Methods The clinical data (including the signs and symptoms,imaging studies,tumor markers,treatment modalities and histopatbologic diagnoses) of 10 cases of bilateral testicular tumor from January 1980 to December 2004 were reviewed.Their age ranged from 19 to 58 years(mean,34 years).Of the 10 cases,8 with metachronous and 2 with synchronous testicular tumors were identified.The clinical stages at the primary and secondary tumor diagnosis were:5 cases of stageⅠ,3 of stageⅡ;and 6 cases of stageⅠ,1 of stageⅡ,and 1 of stageⅢ,respectively,in 8 metachronous tumor patients.Two syn- chronous tumor patients were both identified as stageⅠdisease.Histological examination showed the primary tumor (seminoma) in 4 cases and the secondary contralateral tumor (seminoma) in 3.Results Two syn- chronous tumor patients underwent bilateral radical orchiectomy simultaneously,and 8 underwent orchiectomy successively.Retroperitoneal lymph node dissection was performed in 3 cases.Postoperatively,hypogonadism occurred in 10 patients,and 7 of them received androgen replacement therapy.Follow-up ranged from 9 month to 23 years with a mean of 10.5 years.Two patients died of the disease;2 had metastasis (1 of them was alive with metastasis);2 had recurrences and underwent local resection.Conclusions Metachronous bilateral testicular cancers are more common than synchronous bilateral testicular cancers.Seminoma was the most common histopathologic type.Testis-sparing surgery can be performed in selected cases.

OBJECTIVETo observe the change of activation and proliferation ability of rat T-lymphocytes after suppress ICOS gene expression by RNA interference.

METHODSFour interference sites targeting at rat ICOS gene were designed and four pairs of oligonucleotide fragments were cloned into the pSilencer 4.1-CMV neo plasmid vectors then transfected into rat lymphocytes with cationic liposome. The expression of mRNA and protein of ICOS was detected by RT-PCR and flow cytometry. The alteration of lymphocyte proliferation ability was evaluated by mix lymphocyte reaction, and the secretion levels of IFN-gamma and IL-4 were measured by ELISA procedure.

RESULTSAfter transfection, the expression of mRNA and protein of ICOS in test groups were lower than that in control groups (P < 0.05). The ability of T-lymphocytes in proliferation was poor and the levels of IFN-gamma and IL-4 were reduced with ICOS gene shut down.

CONCLUSIONSRNA interference plasmid vector can suppress ICOS expression in rat T-lymphocytes significantly, and it may be useful for further study on transplantation immunity tolerance.

Animals ; Antigens, Differentiation, T-Lymphocyte ; genetics ; metabolism ; Cell Proliferation ; Cells, Cultured ; Female ; Genetic Vectors ; Inducible T-Cell Co-Stimulator Protein ; Interferon-gamma ; metabolism ; Interleukin-4 ; metabolism ; Lymphocyte Activation ; Male ; RNA Interference ; Rats ; Rats, Sprague-Dawley ; T-Lymphocytes ; immunology ; metabolism ; Transfection

OBJECTIVETo investigate the immunological function parameters in patients undergoing dental restoration of nickel-chromium (Ni-Cr).

METHODSSeven hundred and ninety-five Ni-Cr alloy consumers as exposure group, together with 198 controls, were surveyed by the questionnaire and the biological examination of immunological function.

RESULTSAfter splitting all subjects into three groups of equal sample size by urinary Ni or urinary Cr, serum interleukin-1beta(IL-1β) was found to be significantly higher in the group of urinary Ni > 115.73 µg/mol creatinine comparing to the group of urinary Ni < 37.28 µg/mol creatinine (P < 0.05). No changes of immunological parameters in surm [tumor necrosis factor-alpha (TNF-α), IL-1β, and IL-6] were found in other groups of urinary Ni or Cr. Both one way analysis of variance (ANOVA) and linear regression analysis did not find any association between serum changes of immunological parameters and the parameters of alloy restoration (period, number and the level of metal crown uncovered with porcelain).

CONCLUSIONSThis study did not show positive associations between dental restoration of Ni-Cr alloy and serum levels of TNF-α, IL-1β and IL-6.

Adult ; Age Factors ; Chromium ; urine ; Chromium Alloys ; Cross-Sectional Studies ; Dental Restoration, Permanent ; Female ; Humans ; Interleukin-1beta ; blood ; Interleukin-6 ; blood ; Male ; Middle Aged ; Nickel ; urine ; Random Allocation ; Surveys and Questionnaires ; Tumor Necrosis Factor-alpha ; blood

OBJECTIVETo develop human recombinant neutralizing IgG monoclonal antibodies to hepatitis A virus (HAV) by baculovirus expression system.

METHODSThe heavy and light chain genes of two human-derived neutralizing Fab antibodies to HAV were cloned into baculovirus expression vector Pac-kappa-Fc and Pac-L-Fc, and further expressed in insect cells as IgG antibodies. The IgG products were purified and well characterized.

RESULTSThe baculovirus expressed McAb HAFc16 fully retained the specificity of binding to hepatitis A virus and the competition with mouse anti-hepatitis A virus McAb using ELISA. The viral neutralization assay in vitro demonstrated the retention of antibody function after expression of the human antibody in insect cells. The other expressed antibody HAFc78 also has the neutralizing activity but it is directed against different epitopes of HAV when compared with HAFc16.

CONCLUSIONThe recombinant baculovirus/insect cells expressed human neutralizing IgG antibodies to hepatitis A virus retained all biological functions specific for hepatitis A virus. The results provided the possibility of using these antibodies to rapidly protect high risk or early exposure populations from hepatitis A virus infection.

Antibodies, Monoclonal ; biosynthesis ; immunology ; Baculoviridae ; genetics ; Hepatitis A virus ; immunology ; Hepatitis Antibodies ; biosynthesis ; immunology ; Humans ; Immunoglobulin Fab Fragments ; biosynthesis ; immunology ; Immunoglobulin G ; biosynthesis ; immunology ; Immunoglobulin Heavy Chains ; genetics ; Immunoglobulin Light Chains ; genetics ; Recombinant Proteins ; biosynthesis ; immunology

OBJECTIVETo investigate the gene expression of matrix metalloproteinases (MMP-2, MMP-9) and tissue inhibitor of metalloproteinase-1 (TIMP-1) in proliferative and mature hypertrophic scars.

METHODSTotal RNA from 8 normal skin samples and from 16 human hypertrophic scar samples of different maturing stage was respectively extracted, and then mRNA was isolated. The gene expressions of MMP-2, MMP-9 and TIMP-1 in these samples were examined with reverse transcription-polymerase chain reaction (RT-PCR).

RESULTSThe gray scale ratio of MMP-2, MMP-9 and TIMP-1 transcription in normal skin were (3.8 +/- 0.7)%, (5.8 +/-4.4)%, (30.3 +/- 3.0)%, respectively, which were obviously higher than those in proliferative hypertrophic scar [(14 +/- 5)%, (18 +/- 5)%, (38 +/- 4)%, P < 0.05]. The expression of MMP-2 and MMP-9 genes in mature hypotrophic scar returned to normal level, but that of TIMP-1 remained high when compared with that of normal level (P < 0. 05).

CONCLUSIONThe increase in MMP-2, MMP-9 and TIMP-1 gene expression might be involved in the formation of hypertrophic scars, while the lowering of MMP-2 and MMP-9 gene expression might be associated with the maturation of hypertrophic scars.

Cicatrix, Hypertrophic ; genetics ; metabolism ; pathology ; Female ; Gene Expression ; Humans ; Male ; Matrix Metalloproteinase 2 ; genetics ; metabolism ; Matrix Metalloproteinase 9 ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Skin ; metabolism ; pathology ; Tissue Inhibitor of Metalloproteinase-1 ; genetics ; metabolism

This paper aims to investigate if the dental restoration of nickel-chromium based alloy (Ni-Cr) leads to the enhanced excretions of Ni and Cr in urine. Seven hundred and ninety-five patients in a dental hospital had single or multiple Ni-Cr alloy restoration recently and 198 controls were recruited to collect information on dental restoration by questionnaire and clinical examination. Urinary concentrations of Ni and Cr from each subject were measure by graphite furnace atomic absorption spectrometry. Compared to the control group, the urinary level of Ni was significantly higher in the patient group of <1 month of the restoration duration, among which higher Ni excretions were found in those with either a higher number of teeth replaced by dental alloys or a higher index of metal crown not covered with the porcelain. Urinary levels of Cr were significantly higher in the three patient groups of <1, 1 to <3 and 3 to <6 months, especially in those with a higher metal crown exposure index. Linear curve estimations showed better relationships between urinary Ni and Cr in patients within 6-month groups. Our data suggested significant increased excretions of urinary Ni and Cr after dental restoration. Potential short- and long-term effects of Ni-Cr alloy restoration need to be investigated.
Adult ; Chromium ; urine ; Chromium Alloys ; chemistry ; Creatinine ; urine ; Crowns ; Dental Porcelain ; chemistry ; Female ; Humans ; Male ; Metal Ceramic Alloys ; chemistry ; Middle Aged ; Nickel ; urine ; Spectrophotometry, Atomic ; Surface Properties ; Time Factors