Objective To study the relationship between cytokine gene polymorphisms and chronic rejection in kidney allograft recipients and their donors.Methods The cytokine genotypes in- cluding TNF-?,IL-6,IL-10,TGF-?_1 and IFN-?in 144 consecutive first cadaveric kidney allograft re- cipients and 65 corresponding donors were detected by PCR-SSP method.The recipients were followed up for 5 years.The relationship between the TGF-?_1 genotypes of the recipients and donors and the chronic renal allograft rejection was analyzed.Results The incidence of chronic rejection in the recipi- ents with TGF-?_1 high producer genotype(39/91,42.9%)was significantly higher than in those with TGF-?_1 middle or low producer genotype(9/53,17.0%,P0.05).The incidence of chronic rejection in the recipients and donors with TGF-?_1 high producer genotype(4/4)was significantly higher than in those with other genotypes(31/110,28.2%,P

BACKGROUNDSuicide gene therapy is a widely used molecular treatment for head and neck cancer. In this study, we try to use the method of homogenous recombination in bacteria to clone thymidine kinase gene (tk)-a kind of suicide gene to adenovirus backbone vectors for the construction of replication-defective adenoviruses.

METHODSpAdTrack-CMV/tk was constructed through subclone of a restriction endonuclease fragment including thymidine kinase gene from plasmid pCMV-tk to another plasmid pAdTrack-CMV, and then co-transfected with supercoiled pAdEasy-1, which was an adenoviral backbone vector except for deletions of E1 and E3, to competent E. coli BJ5183 for homogenous recombination using electroporation procedure. With the same method, pAdTrack-CMV was also co-transformed with pAdEasy-1 for homogenous recombination in BJ5183. Identified with restriction endonuclease PacI and polymerase chain reaction (PCR), plasmids pAd-GFP/tk and pAd-GFP were successfully constructed. Each of them was digested with PacI and sequently transfected into human embryo kidney 293 cells (HEK293) using Lipofectamine 2000.

RESULTSComet-like adenovirus-producing foci of Ad-GFP/tk and Ad-GFP were observed after 5 to 7 days of cell culture. After twelve days of packaging, the replication-defective adenoviruses were collected. Identified with PCR, thymidine kinase gene was successfully constructed into Ad-GFP/tk.

CONCLUSIONThe replication-defective adenoviruses containing thymidine kinase can be constructed more easily by homogenous recombination in bacteria than conventional techniques.

Adenoviridae ; genetics ; Bacteria ; genetics ; Defective Viruses ; genetics ; Genetic Therapy ; methods ; Green Fluorescent Proteins ; genetics ; Recombination, Genetic ; Thymidine Kinase ; genetics ; Virus Replication

Objective To evaluated the clinical results of treatment of osteonecrosis of the femoral head(ONFH)by using bone flap pedicled with gluteal muscle branch of ascending branch of the lateral femo- ral circumflex artery and transversal branch of the lateral femoral cireumflex artery.Methods Bone flap pedicled with double blood vessels was applied in 32 hips with stageⅡandⅢONFH according to the Fieat staging system.Fifteen hips were in stageⅡand 17 were in stageⅢ.All patients were followed up for a mean of 25 months(range,15-38 months)and were assessed clinically and radiologically according to Harris sco- ring.Selective digital subtraction angiography was performed in 9 cases to confirm the blood reconstruction of femoral head.Results Two cases underwent total hip replacement postoperatively.Preoperative and postop- erative Harris score were 55 and 88.6.Clinical success rate was 90.6% and radiological success rate was 87.5%.DSA demonstrated blood supply was found reconstruction in the necrotic femoral head.Conclusion Transplantation of bone flap pedicled with gluteal muscle branch of ascending branch of the lateral femoral circumflex artery and transversal branch of the lateral femoral circumflex artery appears to be a effective treat- ment for osteonecrosis of the femoral head,blood supply of bone flap and blood reconstruction of femoral head was confirmed.Early clinical results were satisfied.

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 research on the conservative methods used to preserve the femoral head of patients with osteonecrosis of the femoral head.

METHODSIn these series surgical procedures the osteonecrotic lesion was removed and various vascularized bone blocks or periosteal flaps with its nutrient vessels were transferred to regain sphericity of the femoral head and reinforce the sequestrum. The current study assessed 1005 patients (1226 hips) operated on from 1989 to 2002 with an average follow-up of 5.1 years (range, 1.5-15 years). The mean age of the patients was 37.4 years (range, 17-65 years).

RESULTSSixty-one hips (57 patients) had conversion surgery to a total hip arthroplasty because of progressive collapse or severe pain, or both. In the patients without failure, postoperative Harris hip score improved significantly. Of the 1174 reconstructions that were in situ, 1041 (89.4%) were clinically successful, and 878 (75.4%) were radiologically successful. In relation to the stage of necrosis according to the classification system of Ficat and Alert, good results were achieved in 95.3% of the patients with stages II disease, 87.9% with stages III and 60.8% with stages IV.

CONCLUSIONSConservative methods of vascularized bone block or periosteal flap transfer should be considered in active symptomatic patients to preserve the femoral head. In addition, the earlier the stages of the disease the better outcome could be obtained.

Adolescent ; Adult ; Aged ; Bone Transplantation ; methods ; Female ; Femur ; surgery ; Femur Head Necrosis ; surgery ; Follow-Up Studies ; Humans ; Male ; Microsurgery ; Middle Aged ; Retrospective Studies

OBJECTIVETo investigate the changes in the activity of Escherichia coli asparaginase (L-asp) and the concentration of asparagines (ASN) in the plasma of the acute lymphoblastic leukemia (ALL) children receiving L-asp containing chemotherapeutic protocol to explore more reasonable usage of L-asp in the treatment of childhood ALL.

METHODSL-asp containing hemotherapy regimen of VDLP was used, in which L-asp (10,000 U/m(2)) was administered intravenously every other day for 10 doses in 15 children with ALL. A total of 340 peripheral blood samples were collected at scheduled time points during the therapy and plasma L-asp activity (by spectrophotometric assay) and asparagines concentration (by RP-HPLC) were measured.

RESULTSDuring the administration of L-asp, the plasma L-asp activity was increasing gradually peaked after eight doses and then decreased gradually, while the plasma concentration of asparagines maintained in complete or nearly complete depletion status. After the therapy courses finished, a plasma L-asp activity above 100 U/L with asparagines almost complete depletion status was lasting for about seven days.

CONCLUSIONThe current L-asp containing chemotherapeutic protocols in which L-asp was administered in a dose of 10 000/m(2) intravenously every other day, are efficient enough for the depletion of plasma ASN.

Adolescent ; Antineoplastic Combined Chemotherapy Protocols ; blood ; pharmacokinetics ; therapeutic use ; Asparaginase ; administration & dosage ; blood ; pharmacokinetics ; Asparagine ; blood ; Child ; Child, Preschool ; Drug Administration Schedule ; Female ; Humans ; Infusions, Intravenous ; Male ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; blood ; drug therapy ; Treatment Outcome

Objective To conduct serological investigation on H5N1/H9N2/H7N7 infection among people occupied in poultry fields. Methods Serum samples were collected from people working in live poultry and none-poultry retailing food markets, poultry wholesaling, large-scale poultry breading factories and in small-scale farms, wide birds breeding, swine slaughtering houses and from normal population. Antibodies of H5, H9 and H7 with hemagglutination inhibition and neutralization tests were tested and analyzed. Logistic regression and χ2 test were used. Results Among 2881 samples, 4 were positive to H5-Ab(0.14%), 146 were positive to H9-Ab (5.07%) and the prevalence of H9 among people from live poultry retailing (14.96%) was the highest. Prevalence rates of H9 were as follows: 8.90% in people working in the large-scale poultry breading factories, 6.69% in the live poultry wholesaling business, 3.75% in the wide birds breeding, 2.40% in the swine slaughtering, 2.21% in the non-poultry retailing, 1.77% in the rural poultry farmers and 2.30% in normal population. None was positive to H7-Ab among 1926 poultry workers. Conclusion The H5 prevalence among people was much lower than expected, but the H9 prevalence was higher. None of the populations tested was found positive to H7-Ab. There was a higher risk of AIV infection in live poultry retailing, wholesaling and large-scale breading businesses, with the risk of live poultry retailing the highest. The longer the service length was, the higher the risk existed.

AIM:To evaluate the clinic efficacy of 1g/L anthocyanin eye-patch for mild and moderate dry eye. METHODS: ln this prospective and multicenter study, a total of 320 cases (640 eyes) with mild and moderate dry eye were enrolled from 8 Aier Eye Hospitals in Changsha, Chongqing, Wuhan and so on from October 2016 to April 2017. The patients were assigned to eye patch group (160 cases) and artificial tears group (160 cases) based on random number table. The patients in eye-patch group used 1g/L of anthocyanin eye-patch for more than 6h during night sleep, while the patients in artificial tears group used polyvinyl alcohol eye drops for 4 times per day. The evaluation of symptoms and signs were conducted on 1d before the treatment and 14d after the treatment. The evaluation of symptoms adopted the Ocular Surface Disease lndex ( OSDI), while the observation of signs included tear secretion test (Schirmer ︳ test, S︳t), first noninvasive tear breakup time (NITBUTf) as well as average noninvasive tear breakup time (NITBUTav) measured by Oculus ocular surface analyser. RESULTS: OSDI score, NITBUTf and NITBUTav in the two groups after treatment were significantly improved compared with that before treatment, and the difference had a statistical significance (P < 0. 05). While the difference of S︳t in the two groups before and after treatment had no statistical significance(P>0.05). There was no significant difference in OSDI score, NITBUTf, NITBUTav and S ︳ t between the two groups after treatment (P>0.05). CONCLUSION: The 1g/L anthocyanin eye-patch has similar efficacy with artificial tears for mild and moderate dry eye, which can effectively improve the symptoms and tear film stability.

OBJECTIVE: To investigate the effect of decellular treatment on the framework constituents of extracellular matrix and tissue stability in bovine jugular vein conduit (BJVC), and to provide an evidence for tissue engineering of vascular prosthesis. METHODS: Bovine jugular veins were obtained fresh from a local slaughterhouse and were stored in chilled PBS. In the laboratory, any fat and loose connective tissue on the outer surface of the vessel was trimmed. BJVCs were decellularized by a 3-step extraction method as detergent Triton X-100 (0.5%), Trypsin (0.025%) EDTA (0.02%), and DNase I(30kU/L) RNaseA(0.3g/L). Histological and transmission electron microscopy (TEM) techniques were used to study the framework constituents of extracellular matrix of treated the examples, and fresh tissues were used as controls. Tissue contents of hydroxyproline(alkaline hydrolysis method) and elastin (Fastin Elastin Assay) were assayed respectively in the fresh and decellularized groups (n=10). The vascular wall heat shrinking temperature and mechanical strength were measured to evaluate the tissue stability (n=10). RESULTS: Histochemical and TEM analysis of BJVCs treated with decellularization proved a complete removal of nuclear and other cell components. Tissue collagen was well kept,but elastin was partly lessened. Tissue content of hydroxyproline increased comparatively [(25.73+/-2.97)mg/g vs. (29.25+/-2.99)mg/g, P<0.05] and the elastin content obviously decreased [(159.71+/-21.06)mg/g vs. (134.91+/-35.40)mg/g, P<0.05] in the decellular treatment group compared with the control group. The heat shrinking temperature and tensile stress of decelluarized tissue were lower than those of the fresh tissue[(72.50+/-0.53) degrees C vs. (69.75+/-0.54)degrees C ,P<0.05], [(5.19+/-0.65)MPa vs. (3.13+/-0.94)MPa, P<0.05]. CONCLUSION The basic framework of extracellular matrix in the decellularized BJVC is partly damaged and tissue stability is reduced. Decellularized BJVC should be further crosslinked before being used as a tissue engineering scaffold for clinical pulmonary artery graft.
Animals ; Blood Vessel Prosthesis ; Cattle ; Extracellular Matrix ; Jugular Veins ; Tissue Engineering ; methods ; Tissue Scaffolds

Adult ; Female ; Humans ; Influenza A Virus, H1N1 Subtype ; immunology ; Influenza Vaccines ; immunology ; Influenza, Human ; prevention & control ; Male ; Middle Aged ; Vaccination