The production conditions of glutathione with shaking flask fermentation by the mutant Saccharomyces J-X25, a methionine-defected strain were studied, and the optimum culture conditions are as follows: initial pH6. 0, temperature 30℃, 100ml in 500ml flask, the inoculum size 10% and agitation rate 220r/min. The emphasis was on the stimulating effect on the cells by dioxogen and the sodium lactate as surfactant. Both of which were added at the logarithmic phase of fermentation, and the GSH production was up to 0. 253g/L , 52% higher than the control without the additions. Compared with the production of GSH initial strain that by the mutant in optimum conditions was increased by 79%.

Functional targets are the objects that Chinese herbal medicines act directly upon. If the relationships between the properties of Chinese herbs and their functional targets were analyzed clearly, it would benefit the overall understanding of the holistic mechanisms of Chinese herbal treatments. In this paper, data regarding the properties of Chinese herbs and their functional targets were collected from the 2005 edition of The People's Republic of China Pharmacopoeia. After analyzing and assessing the data, the relationships were defined between the four qi, meridian entry and medicinal functional targets and between the four qi, five flavors and mode of function. Then the relationships between a single herbal medicine and a prescription were analyzed, and the results conformed with the traditional knowledge of Chinese herbal nature and efficacy. This demonstrated that the holistic mechanisms of the properties of Chinese herbs adhere to the findings, which may be beneficial for the development and compatibility of Chinese herbal medicines.

Objective To study the hemoprotective effects of a rotary magnetic field(RMF)with radiation- injured mice.Methods C57 BL6/J mice were randomly divided into a control group and a magnetic treatment group.The mice received total body irradiation with 7.0 Gy and 6.5 Gy ~(137)Cs?rays.The treatment group was trea- ted with a RMF for one hour at a time,twice a day.The intensity of the RMF was 0.6T.The survival rate was ob- served for 30 days.On day 7,10,14,21,28 after irradiation,the subjects' peripheral blood cells were counted.On day 12 and 16,the number of bone marrow mononuelear cells(BMNCs)was measured and their ability to form granu- locyte-macruphage colony-forming unit(CFU-GM)was assessed.The pathological sectioning of the femur was per- formed and the expression level of bone morphogenetic proteins(BMPs)in the bone marrow were evaluated.Re- sults The RMF treatment increased the survival rate and duration among the irradiated mice and the number of blood cells in their peripheral blood.Also,RMF treatment could increase the number of BMNCs and improve their ability to form CFU-GM on days 12-16.Furthermore,RMF could improve angiogenesis and the expression level of BMPs. Conclusion The RMF treatment had an obvious protective effect against the effects of irradiation,and it accelerated the recovery of hematopoiesis and the hematopoietic microenviroment in mouse bone marrow.

ObjectiveTo study the hemoprotective effects of rotating and stationary magnetic field(RSMF) in mice treated with 5-Fluorouracil(5-FU).MethodsThe BalB/C mice were randomly divided into control group and RSMF-treated group.The mice were injected with 5-FU,the dose were 150,180,210,250 mg/kg respectively. RSMF-treated group was exposed to RSMF for 1 h a time,twice a day during 30 consecutive days,and the magnetic intensity was 0.6 T.The survival rate and survival days during the 30 days were observed.7,10,14,21,28 days after injection,the peripheral blood cells were counted.On day 8,10 and 14,the number of bone marrow mononuclear cells(BMNC) and the forming ability of colony-forming unit-granulocyte/macrophage(CFU-GM) were measured.The pathological section of femur and the expression level of bone morphogenic proteins(BMPs) in bone marrow were evaluated.ResultsRSMF could increase the survival rate and survival days of mice treated with 5-FU,and induce an increase in hemoglobin concentration,white blood cell count(WBC),red blood cell count(RBC) and platelet number.Also,RSMF could increase the number of BMNC and improve the forming ability of CFU-GM on days 8～14.Furthermore,RSMF could improve the bone marrow angiogenesis and the expression level of BMPs.ConclusionRSMF have an obvious protective effect against chemotherapeutic injury,and it can accelerate the recovery of hematopoiesis and hematopoietic microenvironment in mouse bone marrow.

Objective To investigate if immunological factors associated with rheumatoid arthritis (RA) affect the result of human immunodeficiency virus (HIV) screening by electrochemiluminescence immunoassay (ECLIA) and enzyme-linked immunosorbent assay (ELISA).
Methods 100 RA cases were enrolled from January 2012 to February 2013 into this study. HIV screening was conducted with ECLIA detecting both HIV-1 p24 antigen, HIV-1 and HIV-2 antibodies, with ELISA and colloidal gold method detecting HIV-1 and HIV-2 antibodies. The samples producing positive results were submitted to the Center for Disease Control for confirmation using Western blotting method. The antibody titers of rheumatoid factors (RF) including RF-IgG, RF-IgM, RF-IgA, and CCP-IgG were analyzed by ELISA.
Results The HIV positive-rate determined by ECLIA was significantly higher than that by ELISA and colloidal gold method (P<0.01). The false-positive rate of HIV screening was associated with antibody titers of RF-IgG, RF-IgM, RF-IgA, and CCP-IgG in RA (P<0.01).
Conclusion Immunological factors, including RF and anti-CCP antibody, may influence the screening of HIV by ECLIA, producing false-positive result.

Acupuncture Therapy ; Adult ; Eye Movements ; Humans ; Male ; Miller Fisher Syndrome ; physiopathology ; therapy

OBJECTIVETo observe the characteristics of sperm single-stranded DNA breaks (SSB) and double-stranded DNA breaks (DSB) in infertile men, explore the association of DSB with male infertility, and provide a new observation index and idea for the diagnosis and treatment of the disease.

METHODSThis study involved 60 infertile men (infertility group) and 30 normal healthy males with infertile wives (control group). We comparatively analyzed the seminal parameters of the two groups, determined sperm concentration and viability using the computer aided sperm analysis system, measured the sperm survival rate by hypoosmotic swelling (HOS) test, examined sperm morphology by Diff-Quick staining, and detected sperm DNA damage by two-tail comet assay.

RESULTSNine two-tail comet models were established for detecting sperm DNA integrity. Comparisons between the fertility and control groups showed that the sperm DNA fragmentation index (DFI) was (33.8 ± 13.1) vs (16.3 ± 7.9)% (P < 0.01), the SSB-DFI was (19.2 ± 11.4) vs (14.9 ± 7.6)% (P > 0.05), the SSB-DFI/DFI was (56.8 ± 32.4) vs (91.4 ± 27.8)% (P < 0.01), the DSB-DFI was (23.9 +13.4) vs (6.1 ± 2.7)% (P < 0.01), and the DSB-DFI/DFI was (70.8 ± 19.5) vs (37.4 ± 11.3)% (P < 0.01). The optimal cut-off value of DSB-DFI/DFI in the diagnosis of male infertility was 39.5%, with the AUG, sensitivity, and specificity of 0.969, 98.3%, and 90%; that of DSB-DFI was 15.85%, with the AUC, sensitivity, and specificity of 0.912, 86.7%, and 80%; and that of DFI was 18.65%; with the AUC, sensitivity, and specificity of 0.861, 90%, 70%, respectively. In the infertile men, neither SSB-DFI nor SSB-DFI/DFI exhibited any correlation with semen parameters (P > 0.05); DFI was correlated negatively with the percentage of progressively motile sperm, sperm survival rate, and the percentage of morphologically normal sperm (P < 0.05 or P < 0.01), but not correlated with sperm concentration (P > 0.05); both DSB-DFI and DSB-DFI/DFI showed a negative correlation with sperm concentration, sperm survival rate, and the percentages of progressively motile sperm and morphologically normal sperm (P < 0.05 or P < 0.01).

CONCLUSIONDouble-stranded, rather than single-stranded DNA breaks, may be a factor inducing male infertility. The type of sperm DNA strand damage is of much reference value for the assessment of male fertility.

Case-Control Studies ; Comet Assay ; DNA Breaks, Double-Stranded ; DNA Breaks, Single-Stranded ; DNA Fragmentation ; Fertility ; Humans ; Infertility, Male ; diagnosis ; genetics ; Male ; Semen Analysis ; Sensitivity and Specificity ; Sperm Count ; Spermatozoa ; Staining and Labeling

The 1.7 kb full-length hemagglutinin (HA) gene fragment of H5N1 subtype avian influenza virus was amplified by RT-PCR and then cloned into the pFastBacHT donor plasmids. The recombinant plasmid pFastBac-H5 was identified by restriction enzyme digestion and sequencing. Following the transposition pFastBac-H5 into the bacmid in DH10Bac E.coli competent cells, the colonies were identified by blue and white selection. The recombinant bacmid (rBacmid-H5) was verified by PCR analysis. Transfection of rBacmid-H5 DNA into sf9 cells using Cellfectin reagent results in the production of recombinant viral stock. Cells were harvested 72h post infection and analyzed by SDS-PAGE, Western-blot, hemagglutination and hemagglutination inhibition test;the expressed HA protein (rH5)shows hemaggluting activity and can be inhibited by H5N1 virus immunized chicken sera. On the other hand, immunization of chickens with rH5 protein results in high titers of H5N1 virus specific hemagglutation inhibition antibodies, which proved its biological activity.

Objective To investigate the influences of mutation at precore and basic core promoter(BCP) region in hepatitis B virus(HBV) on the immune response of specific cytotoxic T lymphocytes(CTL) in patients with chronic hepatitis B(CHB).Methods The number of specific CTL in peripheral blood mononuclear(PBMC) of CHB patients were tested by cytokine flow cytome- try(CFC) and HBV core18-27 peptide.HBV precore and BCP fragments were directly sequenced. Results Twenty-one(38.9%) samples were HBV precore G1896A mutation.Twenty-six(48.1%) samples were BCP region 1762/1764 combined mutation.Thirteen(24.1%) stains were three sites mutated simultaneously.Stimulated with HBV core 18-27 in vitro,the specific CTL level was signifi- cantly higher in the patients with G1896A mutation and BCP region mutation [(0.41?0.09)%, (0.36?0.08)%,(0.48?0.08)%,respectively]than those without mutation[(0.11?0.06)%, P

AIM: To observe the effect of Ahmed glaucoma valve implantation intravitreal bevacizumab in the treatment of neovascular glaucoma ( NVG) .
METHODS:Twenty-two cases (22 eyes) who presented with NVG were first treated with intravitreal bevacizumab 0. 1mL ( 2. 5mg ), then with Ahmed glaucoma valve implantation after regression of iris neovessels. Cases were followed - up for 6 - 36 ( mean 24 ) mo with observation on visual acuity, IOP control, regression of iris neovessels, and complications during or after surgery.
RESULTS: Iris neovessels was regressed in different degree after injection within 1wk in 22 eyes. At final follow-up, the IOP of 18 eyes were all less than 21mmHg without any drugs and of 3 eyes with 1-3 kinds of anti-glaucoma drugs after combined Ahmed glaucoma valve implantation. The IOP of one eye was controlled after cryotherapy. The mean IOP dropped from 45. 36 ±8.13mmHg preoperatively to 15. 59 ± 3. 21mmHg postoperatively. IOP reduction was statistically significance between preoperative and postoperative ( P<0. 05) at final follow-up. Visual acuity was improved in 9 eyes (41%) and was no changed in 13 eyes. No serious complications were observed during or after intravitreous bevacizumab injection and Ahmed glaucoma valve implantation.
CONCLUSION: Ahmed glaucoma valve implantation and intravitreal bevacizumab in the treatment of NVG is useful and safe. It improves the success rate of surgery and preserves visual function, furthermore its complications are less.