The crude toxin was extracted from hypha and culture solution of Phyllosticta commelimecola through three different polarity solvent: benzinum, puncificatum ethyl acetate and chloroform. The result indicated that the toxin secreted by Phyllosticta commelimecola not only was in hypha but also in culture solution and the extracting effect of ethyl acetate was the best. The soybean median and PSK media can be respectively used as solid and liquid culture media to produce toxin and grow mycelium. The optimal cultural conditions for producing toxin were temperature 32℃,cultured period 14d, cultured ways shaking of 150r/min.

BACKGROUNDIt is known that ultraviolet irradiation can affect cellular function through a number of signaling pathways. (-)-epigallocatechin-3-gallate (EGCG) is the major effective component in green tea and can offer protection from ultraviolet-induced damage. In this study, we investigated the protective mechanism of EGCG on human dermal fibroblasts damaged by ultraviolet A (UVA) in vitro.

METHODSTranscription factor Jun protein levels were measured by Western blot. Matrix metalloproteinase 1 (MMP-1) and tissue inhibitor of metalloproteinase-1 (TIMP-1) mRNA were studied by reverse transcription-polymerase chain reaction (RT-PCR) analysis in conjunction with computer-assisted image analysis. MMP-1 and TIMP-1 proteins were quantified by enzyme-linked immunosorbent assay (ELISA).

RESULTSEGCG decreased transcription activity of Jun protein after induction by UVA. Both the mRNA and protein levels of MMP-1 were increased by UVA irradiation, while no significant changes were observed in TIMP-1 levels. The ratio of MMP-1 to TIMP-1 showed statistically significant differences compared with the control. EGCG decreased the ratio of MMP-1 to TIMP-1 by inhibiting UVA-induced MMP-1 expression (P < 0.05).

CONCLUSIONEGCG can protect human fibroblasts against UVA damage by downregulating the transcription activity of Jun protein and the expression of MMP-1. The ratio of MMP-1 to TIMP-1, rather than the levels of MMP-1 or TIMP-1 alone, may play a significant role in human skin photodamage.

Catechin ; analogs & derivatives ; pharmacology ; Cells, Cultured ; Fibroblasts ; metabolism ; radiation effects ; Gene Expression Regulation ; drug effects ; Humans ; Matrix Metalloproteinase 1 ; biosynthesis ; genetics ; Proto-Oncogene Proteins c-jun ; analysis ; RNA, Messenger ; analysis ; Radiation-Protective Agents ; pharmacology ; Reverse Transcriptase Polymerase Chain Reaction ; Tissue Inhibitor of Metalloproteinase-1 ; biosynthesis ; genetics ; Ultraviolet Rays

BACKGROUNDNitic oxide (NO) has been implicated in the pathogenesis of various inflammatory diseases, including sunburn and pigmentation induced by ultraviolet irradiation. Epigallocatechin-3-gallate (EGCG) is the major effective component in green tea and can protect skin from ultraviolet-induced damage. The purpose of this study was to investigate the protective mechanisms of EGCG on inducible nitric oxide synthase (iNOS) expression and NO generation by ultraviolet B (UVB) irradiation in HaCaT cells.

METHODSHaCaT cells were irradiated with UVB 30 mJ/cm 2 and pretreated with EGCG at varying concentrations. The iNOS mRNA was detected by reverse transcriptase polymerase chain reaction (RT-PCR) and NO production was quantified by spectrophotometric method. The expression of NF-kappaB P65 was measured by immunofluorescence cytochemistry staining.

RESULTSThe expression of iNOS mRNA and generation of NO in HaCaT cells were increased by UVB irradiation. EGCG down regulated the UVB-induced iNOS mRNA synthesis and NO generation in a dose dependent manner. The UVB-induced ctivation and translocation of NF-kappaB were also down regulated by EGCG treatment in HaCaT cells (P < 0.01).

CONCLUSIONSGreen tea derived-EGCG can inhibit and down regulate the UVB-induced activation and translocation of NF-kappaB, expression of iNOS mRNA and generation of NO respectively, indicating EGCG may play a protective role from UVB-induced skin damage.

Catechin ; analogs & derivatives ; pharmacology ; Cells, Cultured ; Gene Expression Regulation, Enzymologic ; drug effects ; Humans ; Keratinocytes ; metabolism ; radiation effects ; Nitric Oxide ; biosynthesis ; Nitric Oxide Synthase Type II ; genetics ; Protein Transport ; drug effects ; RNA, Messenger ; analysis ; Tea ; Transcription Factor RelA ; metabolism ; Ultraviolet Rays ; adverse effects

OBJECTIVETo develop a specific quality of life (QOL) instrument for Chinese patients living with HIV/AIDS and to assess its reliability and validity.

METHODSA primary questionnaire was developed with reference to existing instrument and information through discussion with experts, front-line professionals and patients living with HIV/AIDS. This questionnaire was applied in 443 patients living with HIV/AIDS. Some items were removed through the results of responsive rate, factor analysis, correlation matrix,internal consistency and so on. Reliability and validity were assessed after items reduction.

RESULTSThe developed instrument contained 44 items, including 10 scales and other 3 items used for the evaluation of curative effect and side effect of anti-virus drug. No substantial ceiling/floor effects existed. All scales' internal consistency coefficients were > or = 0.70 and the whole instrument was 0.90. The two week retest reliability coefficient of the whole instrument was 0.80, and there were 6 scales below 0.70 of this coefficient. Multitrait/multiitem assessment indicated scaling success rates were all 100% for all items. Spearman correlation coefficient of total score of this instrument with SF-36 was 0.69.

CONCLUSIONIt was the first time that the developed QOL-CPLWHA instrument introducing the scale of hostility regarding mentality trend as well as expressed its acceptable reliability and validity so could be used for the evaluation of Chinese patients living with HIV/AIDS.

Acquired Immunodeficiency Syndrome ; psychology ; Adolescent ; Adult ; Aged ; China ; Female ; Humans ; Male ; Middle Aged ; Quality of Life ; Reproducibility of Results ; Surveys and Questionnaires ; Young Adult

OBJECTIVETo assess the quality of life (QOL) and hostile mentality trend (HMT) of 299 patients living with HIV/AIDS (Human immunodeficiency virus/Acquired immune deficiency syndrome) in three provinces in China, and to understand the major concerns of the these patients.

METHODSThe SF-36 (short form -36) was used for assessing the QOL among 299 HIV-infected patients in Sichuan, Hubei and Guizhou provinces. Reliability and validity of SF-36 were evaluated. Consulting with experts and professionals, seven additional items were developed to evaluate the HMT. Mean scores of the 8 scales were compared between the patients and general rural residents in Sichuan province.

RESULTSFor SF-36, internal consistent coefficients (Cronbach's alpha) of the 8 scales were between 0.75 to 0.90, test-retest reliability coefficient ranged from 0.54 to 0.80. The item-subscale correlation coefficients ranged from 0.46 to 0.97. Mean scores of the 8 scales of the patients ranged from 28.50 to 77.87, and 70.27 to 91.87 for the general rural residents. The variations of the scales were tested by means of Mann-Whitney test with u value ranged from -17.43 to -23.87. The QOL of the patients living with HIV/AIDS were significantly inferior to those of general population (all P < 0.01). The mean scores of the seven items to evaluate HMT ranged from 46.21 to 82.89. The major concerns of the patients living with HIV/AIDS included financial insecurity and family responsibilities, followed by death threat and no cure of HIV/AIDS.

CONCLUSIONThe SF-36 is a reliable instrument for assessing QOL of patients living with HIV/AIDS. The QOL of the patients living with HIV/AIDS in China is poor.

Acquired Immunodeficiency Syndrome ; psychology ; Adolescent ; Adult ; Aged ; Educational Status ; Female ; HIV Infections ; psychology ; Hostility ; Humans ; Male ; Marital Status ; Middle Aged ; Quality of Life ; Reproducibility of Results ; Surveys and Questionnaires ; standards ; Young Adult

OBJECTIVETo explore the effects of antisense epidermal growth factor receptor (EGF-R) oligodeoxynucleotides on ultraviolet-induced c-jun activity of keratinocytes after EGF-R oligodeoxynucleotides transfect to HaCaT in vitro.

METHODSc-jun DNA binding activity after ultraviolet-B (UVB) irradiation and EGF-R oligodeoxynucleotides transfection were determined with a highly sensitive and specific colorimetric method. After EGF-R oligodeoxynucleotides transfection, the mRNA level of EGF-R was detected by reverse transcription polymerase chain reaction method.

RESULTSCompared with control groups, c-jun activity increased significantly in UVB (10, 20, 30 mJ/cm2) irradiation groups (P < 0.05). EGF-R mRNA and c-jun activities induced by UVB were inhibited after the keratinocytes were transfected with EGF-R antisense oligodeoxynucleotides at 2, 4 and 8 microg/ml concentrations (P < 0.01).

CONCLUSIONThe ultraviolet-induced c-jun activity of keratinocytes can be mediated by EGF-R and inhibited by EGF-R antisense oligodeoxynucleotides, which is transfected to keratinocytes and mediated by lipofectamine.

Cell Line ; Humans ; JNK Mitogen-Activated Protein Kinases ; metabolism ; Keratinocytes ; drug effects ; metabolism ; radiation effects ; Oligodeoxyribonucleotides, Antisense ; pharmacology ; Receptor, Epidermal Growth Factor ; biosynthesis ; genetics ; Transfection ; Ultraviolet Rays

Cerebral hypoxic preconditioning (CHP), which was induced by repetitive sub-lethal hypoxic insult, is an endogenous protection of neuron against subsequent severe hypoxic injury. Although a number of possible induction pathways have been investigated, such as neuroactive cytokines, activation of glutamate receptors, the ATP-sensitive potassium channel, nitric oxide and oxidative stress, the exact mechanism underlying CHP-induced protection remains unclear. It is interesting that all the above-mentioned mechanisms are involved in the activation of protein kinases C (PKC). Recently we reported that the level of PKCs membrane translocation was significantly increased in the brain of hypoxic preconditioned mice. In order to explore the role of conventional protein kinases C (cPKC) in the development of cerebral hypoxic preconditioning, biochemical techniques of SDS-PAGE and Western bolt were applied to observe the effects of repetitive hypoxic exposure (H1-H4) on the level of cPKCalpha and gamma membrane translocation in the cortex and hippocampus of mice. Experiments were carried out in accordance with the National Institutes of Health guide for the care and use of laboratory animals. The hypoxic preconditioned mice model was adapted with minor modification from our previous report. In brief, healthy adult BALB/C mice weighing 18-20 g of either sex were randomly divided into 5 groups: control group (H0), hypoxic control group (H1, hypoxic exposure once ), hypoxic preconditioned group (H2-H4, repetitive hypoxic exposure for 2-4 times respectively). The first sign of gasping breath was taken as the end of each hypoxic exposure, and then the mice were kept in normal control condition for a 30-min interval to recover before the following hypoxic insult. We found that the level of cPKCgamma membrane translocation was increased significantly (*P<0.05, n=6) with the increase of the hypoxic exposure times in both hippocampus (H0: 100% vs H1 approximately H4: 119.2%+/-7.0% *, 139.3% +/-7.4%*, 134.2% +/-8.95%*, 184.0% +/-10.8%*) and cortex (H0: 100% vs H1-H4: 129.7% +/-13.8%, 143.3% +/-13.9%*, 204.0% +/-12.1%*, 229.5% +/-14.6%*) of mice. But there were no significant changes in cPKCalpha membrane translocation in cortex and hippocampi of hypoxic preconditioned mice. These results suggest that cPKCgamma plays an important role in the development of cerebral hypoxic preconditioning. The changes in some other forms of novel and atypical PKCs are still under investigation.
Animals ; Brain ; blood supply ; metabolism ; physiology ; Cell Membrane ; metabolism ; Female ; Hippocampus ; metabolism ; Ischemic Preconditioning ; Male ; Mice ; Mice, Inbred BALB C ; Protein Kinase C ; metabolism ; Protein Transport

AIM:To investigate the myopia of junior high school students in Enshi, Hubei province with different selenium content, and analyze the correlation between the level of serum selenium, hair selenium and myopia.METHODS:A cross-sectional study. A total of 600 students from grades 1-3 of junior high schools(100 students in each grade)in selenium-rich(selenium in soil ≥1.28mg/kg)and selenium-deficient(selenium in soil <1.28mg/kg)areas were randomly selected from September 2020 to September 2021, respectively. The level of serum selenium, hair selenium, glutathione peroxidase(GSH-Px)and myopia condition were determined.RESULTS:The serum and hair selenium of myopic group(n=244, 40.7%)were 75.14±11.16μg/L and 0.51±0.01μg/g, respectively. Those in the non-myopic group(n=356, 59.3%)were 110.24±12.14μg/L and 0.68±0.02 μg/g, respectively. The serum selenium and hair selenium in the two groups were different(all P<0.01). The serum selenium of 300 students in the selenium-deficient area was 76.74±11.25μg/L, the hair selenium was 0.45±0.01 μg/g, and the number of myopia cases was 154(51.3%); The serum selenium of 300 students in selenium-rich areas was 102.31±10.26 μg/L, the hair selenium was 0.71±0.02 μg/g, and the number of myopia cases was 90(30.0%), the serum and hair selenium in the selenium-rich areas were significantly higher than those compared with the students in selenium-deficient areas, and the myopic incidence was significantly reduced(all P<0.01). The level of GSH-Px of the two areas was 114.65±12.12U/L vs 75.34±13.20U/L(Z=37.994, P<0.01). There is a negative correlation between serum and hair selenium and the myopic incidence(r=-0.542, -0.621, P<0.05).CONCLUSION:Serum and hair selenium is significantly associated with myopia of junior high school students in Enshi, which may provide new ideas for the clinical prevention and treatment of myopia.