Biomedical Research ; China ; Freeze Drying ; Hepatitis A Vaccines ; adverse effects ; immunology ; Humans ; Safety ; Vaccines, Attenuated ; adverse effects ; immunology

Objective To survey the frequency of H deficient phenotype in blood donor population and analyze the serological and genetic characteristics of these individuals.Methods The H deficient phenotype was screened with anti-H monoclonal antibody.The ABO type was screened with serological method and with sequence specific primer polymerase chain reaction(PCR-SSP).FUT1 and FUT2 gene sequences were analyzed with direct sequencing of PCR products and gene cloning products.Result Of 85 390 blood donors,ten individuals were identified to be para-Bombay phenotype.Four h alleles were found in 14 para-Bombay phenotype individuals,h1(nt547-552?ag),h2(nt880-882?tt),h3(nt658c→t),and h_(new-2)(nt328g→a).The FUT1 genotypes of these para-Bombay individuals were h1/h1(6 individuals),h1/h2(7 individuals) and h3/h_(new2)(1 individual),and the frequency of 4 allele were 67.85%(h1),25%(h2),3.57%(h3),and 3.57%(h_(new-2)),respectively.FUT2 gene was analyzed in 12 para-Bombay phenotype individuals,and a mutation of nt357c→t was detected in all FUT2 gene,another mutation of nt716g→a were heterozygous in 5 individuals with h1/h2 genotype.No null FUT2 gene was detected.In serological analysis,all atypical anti-A or anti-B antibody of 14 para-Bombay individuals were inactive at 37℃,7 individuals had active anti-H antibody at 37℃.Conclusion The frequency of H deficient phenotype in Fujian population is about 1:8 500.The h1 and h2 alleles are predominant in Fujian H deficient individuals on h1-Se~(357) and h2-Se~(357,716) haplotype background.

Objective To retrospectively study revisions for infected total hip replacements in 30 cases and discuss the bacteriological characters of the infected total hip replacements,difficulties and strategies in the revision.Methods Thirty revisions of infected total hip replacements were reviewed retrospectively.There were 12 males and 18 females,with mean age of 62.5 years(31-86 years)at revi- sion surgery.Infection was presented one month to four years(mean seven months)after THA operation. The diseases for initial operation included femoral neck fractures in 12 cases,femoral head necrosis in 11,hip osteoarthritis in five and rheumatoid arthritis in two.Twelve eases were treated by one-stage revi- sion and 18 by two-stage revision.Results Before the revision operation,the hip infection were diag- nosed by bacterial culture in 18 cases including five with Staphylococcus epidermidis,four with Staphylo- coccus aureus and nine with other bacteria.Bacteria growth appeared on the specimens from 23 hip joints during the revision surgery but not on the specimens from seven hip joints.Of 12 one-stage revisions,10 cases were followed for mean 16 months,which showed infection recurrence in two eases.Of 18 two-stage revisions,13 cases were followed for mean 20 months,which showed one case with infection recurrence. The mean Harris hip score was improved from preoperative 44 to 84 at follow-up.Conclusions 1) The main bacteria in the infected hip are antibiotic resistant Staphylococcus.2)Because the revision op- eration is difficult,careful preparation before revision is important for success.The fresh surgeon should not attempt.3)The revision strategies should vary according to specific status of the cases.The infection recurrence rate is lower when using a two-stage revision strategy.4)Application of antibiotic bone cement can help improve treatment effect and facilitate functional recovery of the joints.5)The scientific rehabil- itation after operation is very important to functional recovery.

Objective To report the experience of the application of microsurgery in joint replace- ment.Methods There were 22 cases,10 cases with segmental acetabular defects treated with the pedicle sartorius muscle iliac bone grafting,5 cases with vascular repair following major vascular injury of extremity during operation,6 cases with neural repair following neural injury during operation,1 case with serious injury reconstruction by elbow joint replacement and free flap.Results The operations succeeded in 22 cases without any postoperative infection.The mean follow-up was 40.1 months (3-72 months) in 22 cases,in which the joint function improved and the operative result was satisfactory with no joint pain.Conclusion Microsurgical technique can reconstruct bone and tissue defect effectively in joint replacement.

OBJECTIVETo explore the functional effects of MAPK pathway in the pathogenesis of human osteosarcoma.

METHODSGene microarray (Human Genome U133A, Affymetrix) was used to screen the differential expression of genes involved in MAPK pathway between osteosarcoma cell lines and 3 osteoblastic cell lines. KEGG metabolic pathway analysis was performed among significantly increased or decreased genes using the MATLAB software. Immunohistochemical technique was used to detect the expressions of ERK1/2, JNK and p38 proteins among 48 osteosarcoma and benign 24 osteoblastic tumor samples.

RESULTSUsing an entrance limit of > or = 2.0, 18 differentially expressed MAPK pathway-related genes were selected (10 up-regulated, 8 down-regulated) to mapped to the MAPK pathway of KEGG which are all important node genes. The positive rates of ERK1/2, JNK and p38 proteins were 83.3% (40/48), 72.9% (35/48) and 85.4% (41/48) in osteosarcomas,and 12.5% (3/24), 8.3% (2/24) and 16.7% (4/24) in the control group, respectively. The positive rates and expression intensities were statistically different between the 2 groups (P<0.01).

CONCLUSIONMAPK pathway plays an important role in the pathogenesis of osteosarcoma. ERK, JNK and p38 form an intercoordinating network and regulate the cell proliferation, differentiation, apoptosis, invasion and migration in osteosarcoma.

Adolescent ; Adult ; Aged ; Bone Neoplasms ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Child ; Female ; Gene Expression Profiling ; Humans ; JNK Mitogen-Activated Protein Kinases ; metabolism ; Male ; Middle Aged ; Mitogen-Activated Protein Kinase 1 ; metabolism ; Mitogen-Activated Protein Kinase 3 ; metabolism ; Mitogen-Activated Protein Kinases ; metabolism ; Oligonucleotide Array Sequence Analysis ; Osteoblastoma ; genetics ; metabolism ; pathology ; Osteosarcoma ; genetics ; metabolism ; pathology ; Signal Transduction ; Young Adult ; p38 Mitogen-Activated Protein Kinases ; metabolism

Adolescent ; Adult ; Child ; Female ; Hemostasis, Surgical ; methods ; Humans ; Male ; Middle Aged ; Subclavian Artery ; injuries ; Subclavian Vein ; injuries ; Vascular Surgical Procedures ; methods ; Wounds and Injuries ; surgery

OBJECTIVETo explore in-vivo targeted imaging techniques for liver cancer detection using quantum dots (QDs) labeled probes in a nude mouse model of human hepatocellular carcinoma.

METHODSMercaptoacetic acid (MAA) modified QDs were linked to mouse-anti-human alpha-fetoprotein (AFP) monoclonal antibody to form water soluble QD-AFP-Ab probes, which were validated by spectra analyses and transmission electron microscope. The probes were firstly used to detect AFP antigen in human hepatocellular carcinoma cell line HCCLM6 in-vitro by one-step immunofluorescence method. In-vivo tumor xenografts and lung metastases models were then established by inoculation of HCCLM6 cells subcutaneously and into the tail vein of nude mice, respectively. QD-AFP-Ab probes were injected into the tail vein of the tumor bearing mice for live animal fluorescence imaging. Spectra of tumor and normal tissue were analyzed under illumination of Ti: sapphire laser. Serum levels of alanine amino transferase, aspartate amino transferase, blood urea nitrogen and creatinine were determined by conventional biochemical analysis. The liver, spleen, lungs, kidneys, heart and brain of the experimental nude mice were investigated for nonspecific uptake of the probes by confocal microscope.

RESULTSThe QD-AFP-Ab probes had broad excitation spectra and high fluorescence intensity. They could specifically and efficiently recognize AFP antigen in hepatocellular carcinoma cells. Tumor targeting imaging using these probes were successful without any acute toxicity to the experimental animals. Spectra analysis showed that the probes per field were lower in the centre than the periphery of the tumor. Non-specific uptake of QD-AFP-Ab probes occurred mainly in the liver, spleen and lungs.

CONCLUSIONSQD-AFP-Ab probes have good optical properties and biocompatibility for in-vivo targeted imaging of hepatocellular carcinoma. Such approach promises to be highly desirable for molecular targeted research of liver cancer.

Animals ; Antibodies, Monoclonal ; metabolism ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Line, Tumor ; Diagnostic Imaging ; methods ; Fluorescent Antibody Technique ; methods ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Lung Neoplasms ; metabolism ; secondary ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Microscopy, Fluorescence ; Molecular Probes ; metabolism ; pharmacokinetics ; toxicity ; Neoplasm Transplantation ; Quantum Dots ; Tissue Distribution ; alpha-Fetoproteins ; immunology ; metabolism

BACKGROUNDDengue is currently a significant global health problem but no vaccines are available against the four dengue serotypes virus infections. The development of safe and effective vaccines has been hampered by the requirement of conferring complete protection against all four dengue serotypes and the lack of a convenient animal model. Virus-like particles (VLPs) have emerged as a promising subunit vaccine candidate. One strategy of vaccine development is to produce a tetravalent dengue subunit vaccine by mixing recombinant VLPs, corresponding to all four dengue virus serotypes. Towards this end, this study aimed to establish a Pichia pastoris (P. pastoris) expression system for production of dengue virus type 1 (DENV-1) VLPs and evaluate the humoral and cellular immune response of this particle in mice.

METHODSA recombinant yeast P. pastoris clone containing prM and E genes of DENV-1 was constructed and DENV-1 VLPs expressed by this clone were analyzed by sucrose density gradient centrifugation, Western blotting, and transmission electron microscope. Groups of mice were immunized by these particles plus adjuvant formulations, then mice were tested by ELISA and neutralization assay for humoral immune response, and by lymphocyte proliferation and cytokine production assays for a cellular immune response.

RESULTSOur data demonstrated that recombinant DENV-1 VLPs consisting of prM and E protein were successfully expressed in the yeast P. pastoris. Sera of VLPs immunized mice were shown to contain a high-titer of antibodies and the neutralization assay suggested that those antibodies neutralized virus infection in vitro. Data from the T lymphocyte proliferation assay showed proliferation of T cell, and ELISA found elevated secretion levels of interferon IFN-γ and IL-4.

CONCLUSIONSP. pastoris-expressed DENV-1 VLPs can induce virus neutralizing antibodies and T cell responses in immunized mice. Using P. pastoris to produce VLPs offers a promising and economic strategy for dengue virus vaccine development.

Animals ; Antibodies, Neutralizing ; immunology ; Antibodies, Viral ; immunology ; Dengue Virus ; genetics ; immunology ; metabolism ; Enzyme-Linked Immunosorbent Assay ; Fluorescent Antibody Technique ; Male ; Mice ; Mice, Inbred BALB C ; Pichia ; genetics ; metabolism ; T-Lymphocytes ; immunology

OBJECTIVETo explore the operative indications and operative methods of primary total hip arthroplasty for acetabular fracture and to observe the clinical curative effect.

METHODSWe retrospectively summarized and analyzed the traumatic conditions, fracture types, complications, operative time, operative techniques, and short term curative effect of 11 patients (10 males and 1 female, with a mean age of 42.4 years) with acetabular fracture who underwent primary total hip arthroplasty.

RESULTSThe patients were followed up for 6-45 months (mean=28 months). Their average Harris score of postoperative hip joint was 78.

CONCLUSIONUnder strict mastery of indications, patients with acetabular fracture may undergo primary total hip arthroplasty, but stable acetabular components should be made.

Acetabulum ; diagnostic imaging ; injuries ; Arthroplasty, Replacement, Hip ; methods ; Female ; Fractures, Bone ; diagnostic imaging ; surgery ; Humans ; Male ; Middle Aged ; Radiography

The new coronavirus disease (COVID-19) outbreak has challenged us to take unprecedented steps to bring this pandemic under control. In view of the urgency of this situation, convalescent plasma which was used in previous coronavirus outbreaks has emerged as one of the treatment options in this current pandemic. This is mainly due to the fact that convalescent plasma has been studied in a few case series with promising outcomes. In addition, on-going large clinical trials aimed to further evaluate the effectiveness, safety, and optimal dosage, duration and timing of administration of convalescent plasma are indeed revealing a certain level of promising results. Therefore, this article aims to provide an overview of possible mechanisms of actions of convalescent plasma, its benefits and its level of usage safeness by summarizing the existing evidence on the use of convalescent plasma in COVID-19 patients.