Vitamin C precusor-2-keto-L-gulonic acid can be prepared directly by mixed culture of Ghiconobacter oxy-dans SCB329 and Guconobacter subaxydans SCB110. To obtained its high yield, firstly, the proportion of the two micro- organisms, the ingredients of medium and the initial pH were optimized in shake flaskd, then L9 (34) orthogonal experiment confirmed that urea, C. S. L had high degree statistical meaning. Based on these data, an optimized fermentation media was obta ined: D-Sorbitol 9g, C. S.L1.5g, Urea1.5g, KH2PO40.1g, CaCO30.2g. By-product can be inhabited to the greatest extent and the yield increases by 20%.

OBJECTIVETo construct recombinant adeno-associated virus (rAAV) vector containing angiopoietin-1 (ANG-1) gene and to express the ANG-1 in targeting cells.

METHODSANG-1 cDNA was obtained from human spleen by RT-PCR and was inserted into AAV vectors to form rAAV ANG-1, the virus stocks in high titer were harvested. The rAAVANG-1 and rAAV GFP were transferred into pig mesenchymal stem cells and the expression of ANG-1 was detected by Western blot.

RESULTSThe cloned ANG-1 cDNA was 1515bp in length which was in accordance with that reported previously. Titration of rAAVANG-1 stock was 9 X 10(11)v.g/ml. The expression of ANG-1 gene was detected in transfected cells. Forty-eight hours after rAAV GFP was transfected into mesenchymal stem cells, 55% cells expressed GFP.

CONCLUSIONThe constructed rAAV ANG-1 vector has successfully transfered and expressed in pig mesenchymal stem cells.

Angiopoietin-1 ; biosynthesis ; genetics ; Animals ; DNA, Complementary ; genetics ; Dependovirus ; genetics ; metabolism ; Genetic Vectors ; genetics ; Humans ; Mesenchymal Stromal Cells ; metabolism ; Recombinant Proteins ; biosynthesis ; genetics ; Swine ; Transfection

ITS2 sequence was used as a barcode to identify herbal tea ingredient Plumeria rubra and its adulterants. Genomic DNAs from forty eight samples were extracted, the ITS2 sequences were amplified and sequenced bi-direstionlly, and then assembled and obtained using CodonCode Aligner. The sequences were aligned using ClustalW, the genetic distances were computed by kimura 2-parameter (K2P) model and the Neighbor-joining (NJ) phylogenetic trees were constructed using MEGA5.0. Results showed that the length of ITS2 sequence of P. rubra were 244 bp. The intra-specific genetic distances (0-0. 016 6) were much smaller than inter-specific ones between P. rubra and its adulterants(0.320 8-0.650 4). The NJ tree indicated that P. rubra and its adulterants could be distinguished clearly. Therefore, Using ITS2 barcode can accurately andeffectively distinguish herbal tea ingredient P. rubra from its adulterants, which providesa new molecular method to identify P. rubra and ensure its safety in use.
Apocynaceae ; classification ; genetics ; DNA Barcoding, Taxonomic ; methods ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Drug Contamination ; prevention & control ; Drugs, Chinese Herbal ; chemistry ; classification ; Flowers ; chemistry ; classification ; Molecular Sequence Data ; Phylogeny ; Quality Control

Female ; Humans ; Middle Aged ; Neuroma, Acoustic ; complications ; Subarachnoid Hemorrhage ; etiology

Adolescent ; Adult ; Carrier State ; pathology ; psychology ; Female ; Hepatitis B ; pathology ; psychology ; Hepatitis B virus ; Humans ; Liver ; pathology ; Male ; Middle Aged ; Young Adult

AIM:To investigate the effects of cordycepin on the motor and cognition in Parkinson disease mice induced by 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP).METHODS:C57BL/6 mice were intraperitoneally injected with MPTP at a dose of 30 mg/kg daily for consecutive 8 d to establish the model of Parkinson disease.HE staining was used to observe the cell number in the substantia nigra pars compacta (SNpc) from the mice.Western blot was used to detect the protein level of tyrosine hydroxylase (TH) in substantia nigra (SN).The effects of cordycepin on the motor, emotional change and cognitive behavior of the Parkinson disease mice were examined by open-field test (OFT) , spontaneous alternating behavior (SAB) and water maze test (WMT) , respectively.RESULTS:Cordycepin significantly reduced the apoptosis of cells in SNpc and reversed the decrease in the expression of TH in SN induced by MPTP (P<0.05).Furthermore, cordycepin was able to improve the average speed in OFT (P<0.05) , and increased the total number of arm entry and the accuracy in SAB (P<0.05) , but had no obvious effect on the latency in WMT.CONCLU-SION:Cordycepin is capable of attenuating the impairments of motor and explorative ability in the early stage of Parkinson disease mice, but does not alter the cognitive dysfunction.

OBJECTIVETo study the effects of combination of angiopoietin-1 (ANG-1) and vascular endothelial growth factor₁₆₅ (VEGF₁₆₅) gene transfer mediated by recombinant adeno-associated viral vector on the neovascularization in chronic ischemic porcine myocardium.

METHODSAn ameroid constrictor was implanted around the left circumflex coronary artery (LCX) via endoscopy. Six weeks later, coronary angiography revealed that the myocardial ischemia was established by gradual occlusion of the left circumflex coronary artery (LCX). Sixteen swine with the total occlusion or partial stenosis (> 85 %) of the LCX were divided into 4 groups (4 in each group): group I, group II and group IV (control) received direct myocardium injection of rAAV₂ VEGF₁₆₅, rAAV₂ ANG-1 or PBS alone, respectively; group III received rAAV₂ VEGF₁₆₅ and rAAV₂ ANG-1. Selective coronary angiography and ultrasonography were performed perioperatively to evaluate the cardiac function and the formation of collateral circulation. The expression of VEGF₁₆₅ and ANG-1 proteins were assessed using ELISA or Western blot. The degree of angiogenesis was assessed by use of immunohistochemical analysis.

RESULTAngiography showed that the occlusion of all LCX was completed or exceeded 95% 6 weeks after ameroid constrictor implantation, indicating the successful establishment of animal model. The expression levels of VEGF₁₆₅ in group I and III and ANG-1 in groups II and III began to increase at d7 after transfection and reached the peak at d14; then decreased gradually to the normal level after 3 months. The expression levels of VEGF₁₆₅ in group II and group IV or that of ANG-1 protein in group I and group IV had no markedly changes at different time after transfection. There were significant increase in capillary density and arteriole density and more side branch vessels formed in group III compared with other groups. Echocardiographic measurements showed that the left ventricular systolic function of animals in groups I, II and III increased significantly after gene transfection, especially in group III; but there was no changes in group IV.

CONCLUSIONMyocardial perfusion and the left ventricular systolic function are improved after rAAV₂ VEGF₁₆₅ or rAAV₂ ANG-1 transfection, which is associated with the angiogenesis in porcine model of chronic myocardial ischemia.

Adenoviridae ; genetics ; Angiopoietin-1 ; genetics ; Animals ; Collateral Circulation ; Coronary Vessels ; physiopathology ; Disease Models, Animal ; Genetic Therapy ; Genetic Vectors ; Male ; Myocardial Ischemia ; physiopathology ; therapy ; Neovascularization, Physiologic ; Swine ; Swine, Miniature ; Transfection ; Vascular Endothelial Growth Factor A ; genetics

OBJECTIVETo evaluate the effectiveness and safety of sirolimus-eluting stents (SESs) for treatment of in-stent restenosis (ISR).

METHODSAll 27 patients with ISR and clinical evidence of ischemia had been treated with SESs. Among them, 23 patients had diffuse and complex lesions, and 5 of them received 2 SESs. Clinical and angiographic follow-up were performed for all patients and the results were analyzed.

RESULTSAll stents were implanted successfully. There were no remained stenosis and major in-hospital complications. Average follow-up time was 8.9 +/- 2.1 (5-14) months, with a clinical follow-up rate of 96.3% and angiographic follow-up rate of 92.6%. During the follow-up, there was none of death. One patient had recurrent angina with an angiographic evidence of the proximal edge restenosis of the stent. Mild neointimal hyperplasia in the proximal edge was found in 2 patients, but the stenosis was less than 25%. No late lumen loss was found in other 24 patients. The late lumen loss of the in-stent averaged 0.09 +/- 0.02 mm, and of the distal edge vessel averaged 0.10 +/- 0.03 mm, and of the proximal edge vessel averaged 0.20 +/- 0.06 mm. The rate of target vessel revascularization was 3.8%.

CONCLUSIONThe SES implantation is safe and feasible for the treatment of in-stent restenosis, which could effectively prevent neointimal hyperplasia and recurrent restenosis of the lesion.

Adult ; Aged ; Coronary Angiography ; Coronary Restenosis ; diagnostic imaging ; therapy ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Sirolimus ; administration & dosage ; Stents

OBJECTIVETo discuss the treatment of cN(0) tongue carcinoma patients.

METHODS185 cases of the mobile tongue carcinoma patients (male 102, female 83, aged 28 to 88) treated with surgery from 1988.5 to 1995.6 had been followed up and retrospectively analyzed. Extensive resection of the primary tumors and neck dissections were performed, and all the samples were pathological positive.

RESULTSThe cervical lymphatic node metastasis rates for stage I-II, III-IV disease, grade I, II disease were 16.66%, 38.05%, 17.42% and 37.50% respectively. And the rates were 9.00%, 31.37% and 55.55% for submucous infiltration, muscle infiltration and perineural infiltration, respectively. The overall 5 year survival was 72.43%, and the 5 year specific survival rate was 44.44% and 83.96% for those having or not having cervical node metastasis. The levels of 29 patients with positive node metastasis for 148 cN(0) patients were submandibular and submental lymphatic nodes (22.64%), superior deep cervical lymphatic nodes (35.84%), middle deep cervical lymphatic nodes (26.41%), inferior deep cervical lymphatic nodes (15.09%), posterior neck lymphatic nodes (0.00%). The over all 5 year survival rates for selective neck dissection were 85.13% and 21.62% in therapeutic dissection (chi(2) = 29.73, P < 0.01). Patients performed selective neck dissection the rates were 68.96% and 89.07% respectively with or without lymphatic node metastasis. Comparably the 5-year rate was only 20.00% for the patients performed the therapeutic dissection with lymphatic node metastasis.

CONCLUSIONS(1) cN(0) patients should be observed carefully in stage I, and the selective dissection must be performed in stage II-IV. (2) Supraomohyoid ND is essential for T(2) patients, and functional ND is essential for T(3 - 4) patients. (3) There is correlation between cervical metastasis and the stage, grade or infiltration of tongue cancer (P < 0.05). The prognosis could be expected from these factors.

Adult ; Aged ; Aged, 80 and over ; Carcinoma, Squamous Cell ; mortality ; pathology ; surgery ; Female ; Follow-Up Studies ; Humans ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; Male ; Middle Aged ; Neck Dissection ; Neoplasm Staging ; Prognosis ; Retrospective Studies ; Survival Analysis ; Survival Rate ; Tongue Neoplasms ; mortality ; pathology ; surgery ; Treatment Outcome

OBJECTIVESTo explore the category, quantity and clinical significance of autoantibodies on bone marrow hematopoietic cells in patients with immunorelated cytopenia and evaluate the sensitivity of direct antiglobulin reaction (Coombs test ) of bone marrow mononuclear cells (BMMNC).

METHODSThe category and the positive rate of autoantibodies on bone marrow hematopoietic stem cells, nucleated erythrocytes, granulocytes in 32 patients with uncertain immunorelated cytopenia were investigated by using BMMNC-Coombs test and double immunofluorescence flow cytometry.

RESULTSThe positive rate of autoantibodies on bone marrow hematopoietic cells tested by flow cytometry was 90.63% which was higher than that by BMMNC-Coombs test (50.0%) (p < 0.05). In 29 positive cases, IgG autoantibody accounted for 6.90%, IgM13.8%, IgG+IgA 3.4%, IgG+IgM 31.0%, and IgG+IgM+IgA 44.8%. Of the 29 Patients, 25 (86.2%) with IgG autoantibody, 26 (89.7%) with IgM and 14 (48.3%) with IgA. The patients with IgG autoantibody alone had the lowest hemoglobin levels, and those with IgM autoantibody might have intravascular hemolytic findings. The response time of patients with IgG and IgG+IgM was shorter than that of the other patients. 91.3% of the patients had autoantibodies on bone marrow hematopoietic stem cells and showed pancytopenia, and 50% of the patients had autoantibodies on nucleated erythrocytes and granulocytes. Eleven of 13 patients with negative BMMNC-Coombs tests had autoantibodies on bone marrow hematopoietic stem cells detected by FACS. There was no significant difference of the quantities of the three categories of autoantibodies of nucleated erythrocytes and stem cells. The quantities of IgA on granulocytes were lower than that of IgG and IgM. There was no significant difference between IgG and IgM on granulocytes. The quantity of IgA on hematopoietic stem cells was significantly higher than that on nucleated erythrocytes or granulocytes.

CONCLUSIONSThe sensitivity of double immunofluorescence flow cytometry assay was higher than that of BMMNC-Coombs test for detecting autoantibodies. In immunorelated cytopenia patients, the predominant autoantibody was IgM which could cause intravascular hemolysis, and the second one was IgG which could cause severe anemia. Most immunorelated cytopenia patients had autoantibodies on hematopoietic stem cells and showed pancytopenia. IgA was more easily seen on the hematopoietic stem cells.

Adolescent ; Adult ; Aged ; Autoantibodies ; classification ; metabolism ; Autoimmune Diseases ; immunology ; Bone Marrow Cells ; immunology ; Child ; Coombs Test ; Female ; Flow Cytometry ; Humans ; Immunoglobulin A ; metabolism ; Immunoglobulin G ; metabolism ; Immunoglobulin M ; metabolism ; Male ; Middle Aged ; Pancytopenia ; immunology