Phytic acid is a new substitutable plant-derived antifungal agent for the seedling blight of Pinus sylvestris var. mongolica caused by Fusarium oxysporum.

Phytic acid (PA) is a new substitutable plant-derived antifungal agent; however, few reports have been published regarding its antifungal effects on pathogenic fungi. The present study explored the in vitro antifungal activity of PA against four phytopathogenic fungi and found that PA was the most effective at inhibiting the growth of Fusarium oxysporum. This study aimed to investigate the in vivo and in vitro antifungal activities of PA against the seedling blight of Pinus sylvestris var. mongolica caused by F. oxysporum and to determine its possible mechanism of action. The results showed that PA inhibited spore germination and mycelial growth of F. oxysporum in a concentration-dependent manner and exhibited strong inhibition when its concentration exceeded 1000 mg/L. It mainly destroyed the integrity of the cell membrane, increasing its cell membrane permeability, causing the cell contents to spill out, and impairing fungal growth. In addition, the leakage of intercellular electrolytes and soluble proteins indicated that PA used at its EC20 and EC50 increased the membrane permeability of F. oxysporum. The increase in malondialdehyde and hydrogen peroxide content confirmed that PA treatment at its EC20 and EC50 damaged the cell membrane of the pathogen. Scanning electron microscopy revealed that PA affected the morphology of mycelia, causing them to shrivel, distort, and break. Furthermore, PA significantly reduced the activities of the antioxidant-related enzymes superoxide dismutase and catalase, as well as that of the pathogenicity-related enzymes polygalacturonase, pectin lyase, and endoglucanase (EG) in F. oxysporum (P < 0.05). In particular, EG enzyme activity was maximally inhibited in F. oxysporum treated with PA at its EC50. Moreover, PA significantly inhibited the incidence of disease, and growth indices in Pinus sylvestris var. mongolica seedling blight was determined. In summary, PA has a substantial inhibitory effect on F. oxysporum. Therefore, PA could serve as a new substitutable plant-derived antifungal agent for the seedling blight of P. sylvestris var. mongolica caused by F. oxysporum.

First Report of Black Spot Needle Blight of Pinus sylvestris var. mongolica Litv. Caused by Heterotruncatella spartii in China.

Pinus sylvestris var. mongolica Litv. (Pinales: Pinaceae) is an excellent tree for soil and water conservation in Northeast China. The Honghua'erji area in Inner Mongolia is the "hometown of P. sylvestris var. mongolica", however, in recent years, coniferous diseases of P. sylvestris var. mongolica have frequently occurred here. During the investigation, it was found that some black spot needle blight had been observed in addition to the common blight caused by Sphaeropsis sapinea. From May to September 2020, black spot needle blight was found on hundreds of P. sylvestris var. mongolica trees in four forest farms, and the infection rate among the forests was 24.58 % (n=240). This disease first appeared on the upper part of the needles, and the needles then became withered and gradually showed light black spots, although they remained green. As the disease progressed, the needles eventually died and turned gray with many dark black spots. Fungal isolate named YJ-1 was obtained from infected needles of symptomatic pine trees, and a voucher specimen was deposited in Heilongjiang Province Key Laboratory of Forest Protection. Microscopic observation showed the conidia were 3-septate (4 cells) clavate spindles that measured 23.9 µm (20.8-25.9) × 5.9 µm (4.5-8.2) (n=50). The middle two cells were dark brown, and the septa were darker than the cells. Both apical and basal cells were hyaline. The apical cell had 2-4 appendages (mostly 3), and the basal cell had a truncate base (n=50). The cultural characteristics on potato dextrose agar medium were flat off-white and dense in 3-5 d. At approximately 5-7 d, the reverse side of the colony turned pale to slightly luteous. Superficial black acervuli were distributed in the center of the mature colonies after 10 d. Morphological, cultural and microscopic characteristics observed were similar of Heterotruncatella spartii (basionym: Truncatella spartii) reported by Hlaiem et al (2019). To further identify, total DNA was extracted and the internal transcribed spacer region (ITS-rDNA) was amplified by PCR using the primers ITS1/ITS4 and sequenced for BLASTn analysis and phylogenetic tree construction. The resulting 564 bp sequence (GenBank Accession No. OL662864) had 99.24% (521/525) to H. spartii MFLUCC 15-0537, with bootstrap support of at least 94% using the Neighbor-Joining algorithm by MEGA-X (Felsenstein, 1985). The fungus was identified as H. spartii based on morphology and molecular methods. A pathogenicity test was conducted by preparing a conidial suspension of 2.0 × 107 conidia/mL. The suspension was sprayed onto the needles of 20 pots of annual P. sylvestris ar. mongolica seedlings, and the control was sprayed with sterile water. Then the seedlings were placed in a constant temperature room at 25 °C. After 30 d, typical symptoms appeared on 11 inoculated needles, while the control needles remained symptomless. After 50 d, the re-isolation infection rate reached 66.7 %. The fungus present on the inoculated seedlings was morphologically identical to that originally observed on diseased pines, fulfilling Koch's postulates. The fungus was isolated from Spartium junceum for the first time and designated Truncatella spartii (Senanayake et al, 2015). It was then renamed H. spartii (Liu et al, 2019) and has been reported to infect P. pinea in Tunisia (Hlaiem et al, 2019). To our knowledge, this is the first report of H. spartii causing black spot needle blight on P. sylvestris var. mongolica in China and worldwide.

Inhibitory effects and mechanisms of vanillin on gray mold and black rot of cherry tomatoes.

Vanillin is a natural antimicrobial agent; however, there are few reports on its antifungal effect on postharvest pathogenic fungi. This study aimed to investigate the in vivo and in vitro antifungal activities of vanillin against gray mold (caused by B. cinerea) and black rot (caused by A. alternata) of cherry tomato fruit and to explain its possible mechanism of action. Vanillin strongly inhibits Botrytis cinerea and Alternaria alternata mycelial growth, spore germination, and germ tube elongation in a concentration-dependent manner (P<0.05). In vivo experiments showed that 4000 mg L-1 vanillin treatment inhibited cherry tomato gray mold and black rot occurrence. Besides, intercellular electrolytes, soluble proteins, and soluble sugars leakage indicated that 50 or 100 mg L-1 vanillin treatment increased Botrytis cinerea and Alternaria alternata membrane permeability. The increase of malondialdehyde and hydrogen peroxide contents confirmed that 50 or 100 mg L-1 vanillin treatment damages the pathogen membranes. Importantly, vanillin treatment inhibited the pathogenicity-related enzyme activities of the two pathogens to reduce their infection ability, among them PL enzyme activity in A. alternata was most inhibited, reducing by 94.7 % at 6 h treated with 100 mg L-1 vanillin. The hyphae morphology of the two pathogens changed, the mycelia were severely damaged, and the hyphae surface was deformed, shrunk, or even broken after 100 mg L-1 vanillin treatment. In summary, vanillin had a substantial inhibitory effect on postharvest gray mold and black rot in cherry tomato fruit. Therefore, vanillin can be an effective alternative to prevent and control cherry tomato postharvest diseases.

Sodium pheophorbide a controls cherry tomato gray mold (Botrytis cinerea) by destroying fungal cell structure and enhancing disease resistance-related enzyme activities in fruit.

Sodium pheophorbide a (SPA) is a natural photosensitizer. The present study investigated the antifungal activity and mechanism of SPA against Botrytis cinerea in vitro and in vivo. Its inhibitory effect was studied on the spore germination and mycelial growth of B. cinerea. The effects of SPA on cell wall integrity, cell membrane permeability, and mycelial morphology of B. cinerea were also determined. Additionally, how SPA effected B. cinerea in vivo was evaluated using cherry tomato fruit. The results showed that SPA effectively inhibited the spore germination and mycelial growth of B. cinerea under light conditions (4000 lx). SPA significantly affected both cell wall integrity and cell membrane permeability (P < .05). In addition, SEM analysis suggested that B. cinerea treated with SPA (12.134 mg/mL) showed abnormal mycelial morphology, including atrophy, collapse, flattening, and mycelial wall dissolution. In vivo tests showed that SPA could increase the activities of superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) significantly (P < .05); however, SPA had no significant effect on phenylalanine ammonia lyase (PAL) activity. In short, SPA could destroy the fungal cell structure and enhance disease resistance-related enzyme activity in cherry tomatoes, thereby controlling cherry tomato gray mold.

Transcriptome and cell wall degrading enzyme-related gene analysis of Pestalotiopsis neglecta in response to sodium pheophorbide a.

Sodium pheophorbide a (SPA) is a new alternative fungicide with low toxicity and high efficiency, which has high fungicidal activity against Pestalotiopsis neglecta, a pathogen that causes black spot needle blight of Pinus sylvestris var. mongolica. To utilize SPA for plant disease control, understanding its antifungal mechanism is essential. Six cDNA libraries were constructed from 3 d-old P. neglecta mycelia (three SPA-infected and three untreated groups) and 29,850 expressed genes were obtained by Illumina HiSeq4000 sequencing. Compared with controls, 3268 differentially expressed genes (DEGs) were identified in SPA-treated groups, including 1879 upregulated and 1389 downregulated genes. Most DEGs were involved in the metabolism of amino acids, carbohydrates, and lipids, as well as cell structure and genetic information processing. These findings were further confirmed by decreased conductivity, RNA and protein content, and activities of nicotinamide adenine dinucleotide-dependent malate dehydrogenase, citrate synthase, isocitrate dehydrogenase, and succinate dehydrogenase. Moreover, qRT-PCR verified the reliability of the transcriptome results. After treatment with SPA at different concentrations for 60 min, the expressions of three cell wall degrading enzyme-related genes (PnEG, PnBG, and PnPG) were all suppressed. Overall, this study provided insights into the molecular mechanisms through which SPA inhibits P. neglecta, increasing the possibility of developing SPA into an effective fungicide in the future.

[Expressions of JNK and p-JNK in advanced prostate cancer and their clinical implications].

OBJECTIVE: To investigate the expressions of JNK and p-JNK in advanced prostate cancer (PCa) and benign prostatic hyperplasia (BPH) and their implications. METHODS: Using immunohistochemistry, we detected the expressions of JNK and p-JNK proteins in 40 cases of paraffin wax-embedded PCa and 21 cases of BPH tissues and analyzed their relationships with advanced PCa and BPH as well as with the pathologic features of advanced PCa. RESULTS: Statistically significant differences were not found in the positive expression rate of the JNK protein between BPH and PCa (42.86% vs 52.50%, P>0.05), non-metastatic and metastatic PCa (53.85% vs 51.85%, P >0.05), Gleason ≤7 and Gleason >7 (58.82% vs 47.82%, P >0.05), PSA ≤20 µg/L and PSA >20 µg/L (57.14% vs 51.52%, P >0.05), or survival >5 yr and survival ≤5 yr (60.00% vs 45.00%, P >0.05), nor in the expression level of p-JNK between BPH and PCa (33.33% vs 35.00%, P >0.05), non-metastatic and metastatic PCa (30.77% vs 37.03%, P >0.05), Gleason ≤7 and Gleason >7 (35.29% vs 34.78%, P >0.05), or PSA ≤20 µg/L and PSA >20 µg/L (43.75% vs 10.93%, P >0.05). However, the expression of p-JNK was significantly higher in the survival >5 yr than in the survival ≤5 yr group of the PCa patients (50.00% vs 20.00%, P <0.05). CONCLUSIONS: PCa patients with highly expressed p-JNK have a longer survival time and the high positive rate of p-JNK is associated with the prognosis of PCa.

[Expressions of ERK and p-ERK in advanced prostate cancer].

OBJECTIVE: To investigate the expressions of extracellular signal-regulated kinase (ERK) and p-ERK in benign and malignant prostate tissues, and whether it can be used as a marker for the prognosis of advanced prostate cancer (PCa). METHODS: Using immunohistochemical Envision, we detected the expressions of ERK1/2 and p-ERK1/2 in 20 cases of benign prostatic hyperplasia (BPH) and 40 cases of advanced PCa and analyzed their correlation with PCa metastasis, Gleason score, PSA level, and prognosis. RESULTS: The expression of ERK1/2 was remarkably higher in the advanced PCa than in the BPH cases (82.5% vs 55%, P<0.05), which was not associated with cancer metastasis, Gleason score, PSA level, or survival time of the patients with advanced PCa, and so was that of p-ERK1/2 (75.0% vs 35%, P<0.05), which was not associated with the Gleason score or PSA level of the PCa patients, either. The expression rates of p-ERK in the metastasis, non-metastasis, survival >5 yr, and survival ≤ 5 yr groups were 61.9%, 89.5%, 57.9%, and 90.5%, respectively, with statistically significant differences among these groups (P<0.05). CONCLUSIONS: ERK1/2 and p-ERK1/2 proteins are highly expressed in advanced PCa and p-ERK1/2 is associated with the metastasis and prognosis of advanced PCa.

Antifungal Activity of 6-Methylcoumarin against Valsa mali and Its Possible Mechanism of Action.

Valsa canker of apple (VCA) caused by Valsa mali severely affected apple production in east Asia. With the increase in drug resistance, there is an urgent need for efficient and environmentally friendly antifungal agents. Coumarins have attracted much attention due to their excellent antimicrobial activity against plant pathogens. In this study, the antifungal activity of several coumarins against phytopathogenic fungi was evaluated, and then the antifungal activity of the screened 6-MCM against V. mali and its underlying mechanism was further investigated. The results of the in vitro antifungal activity assay showed that some coumarins had significant inhibitory effects on V. mali. Notably, 400 mg/L of 6-MCM had the best antifungal activity of 94.6%. Further experiments showed that 6-MCM slowed down the growth of V. mali mycelia and the germination of spores in a concentration-dependent manner, with EC50 of 185.49 and 54.62 mg/L, respectively. In addition, 6-MCM treatment increased mycelial conductivity, extracellular protein leakage, and MDA content, resulting in damage to the cell membrane. Moreover, 6-MCM significantly reduced the cell wall degrading enzymes secreted by V. mali, including EG, PG and PL, thereby limiting its pathogenic capacity. SEM and TEM results showed that 6-MCM treatment had a significant effect on the morphology and ultrastructure of mycelial cells. Inoculation of isolated apple branches found that the application of 6-MCM effectively inhibited the development of VCA and significantly reduced the incidence. All these results suggest that 6-MCM has the potential as a green substitute for VCA control.

Incontinentia pigmenti with intracranial arachnoid cyst: A case report.

BACKGROUND: Incontinentia pigmenti (IP) is a rare X-linked dominant genetic disorder that can be fatal in male infants. It is a disease that affects many systems of the human body. In addition to characteristic skin changes, patients may also have pathological features of the eyes, teeth, and central nervous system. Therefore, the lesions in these systems may be the first symptoms for which patients seek treatment. To date, no cases of IP complicated by intracranial arachnoid cyst (IAC) have been reported. This paper aims to report a case of IP with IAC in order to share the diagnosis and treatment experience of this rare case with other clinicians. CASE SUMMARY: An 11-year-old female patient suffered intermittent limb convulsions for five months and was sent to hospital. In the initial stage, the patient was considered to have primary epilepsy. Further investigation of the patient's medical history, physical examination and imaging examination led to the diagnosis of IP combined with intracranial space-occupying lesions, and secondary epilepsy. The patient was treated with craniotomy, and postoperative pathology revealed an IAC. The patient recovered well after craniotomy and had no obvious surgery-related complications. During the follow-up period, the patient did not have recurrent epilepsy symptoms. CONCLUSION: IP is a multi-system disease that presents with typical skin lesions at birth, but the long-term prognosis of this disease depends on the involvement of systems other than the skin, especially nervous system and ocular lesions.

Hypermethylation of estrogen receptor-alpha gene in atheromatosis patients and its correlation with homocysteine.

OBJECTIVE: To investigate the aberrant DNA methylation in promoter region of estrogen receptor alpha (ERalpha) in atherosclerosis (As) and the possible involvement of homocysteine (Hcy) in its pathogenesis. METHODS: The blood samples were collected from 54 patients with As approved by carotid colorized ultrasound examination and 28 healthy control subjects. The methylation status of CpG islands in ER-alpha gene promoter region of genome DNA was analyzed by nested-methylation-specific PCR (nMSP). tHcy was examined by fluorescent-biochemical method. Spearman rank correlation was used to analyse the relationship between the degree of methylation in ER-alpha gene and the level of tHcy. Cultured smooth muscle cells of Homo sapiens were treated by Hcy with different concentrations and different treating time, again the DNA methylation status was assayed by nMSP, and the proliferation of SMC was assayed by MTT. RESULTS: Hypermethylation of ER-alpha gene promoter region was found in 38 cases of atherosclerosis patients, and the methylation frequency was 70.4%. While in healthy controls, just 8 of 28 samples hypermethylation was found, only 28.6% methylation frequency was detected, much lower than the one in atherosclerosis group (p<0.05). Meanwhile, the level of tHcy in atherosclerosis group was significantly higher than that in control group (p<0.05). The spearman rank correlation analysis explored an obvious correlation between the degree of methylation in ER-alpha gene and the level of tHcy (r=0.809, p<0.05), and the severity of atherosclerotic lesion was also heightened along with the increment of plasma level of tHcy. The cultured SMCs treated by Hcy resulted in de novo methylation in promoter region of ERalpha gene with a concentration and treating time-dependent manner, and a dose-dependent promoting effect on SMC proliferation. The in vivo and in vitro data coincidently showed that the Hcy could promote the hypermethylation of ERalpha gene, which may be an important mechanism for the pathogenesis of As. CONCLUSION: Hypermethylation of CpG islands in ER-alpha gene promoter region was found in much higher frequency in atherosclerosis patients, which is positively correlated with the increased level of plasma tHcy and the severity of atherosclerotic lesion, and the in vitro experimental results further extended above clinical data that HHcy can lead to the hypermethylation of ER-alpha gene, and hence to promote the occurrence and development of As.

Intermolecular Dehydrative [4 + 2] Aza-Annulation of N-Arylamides with Alkenes: A Direct and Divergent Entrance to Aza-Heterocycles.

We disclose that following activation with trifluoromethanesulfonyl anhydride, secondary N-arylamides undergo smooth intermolecular dehydrative [4 + 2] aza-annulation with alkenes under mild conditions to give 3,4-dihydroquinolines, amenable to further functionalization. Meanwhile, conditions have been established to allow divergent one pot synthesis of tetrahydroquinolines and quinolines as well as tricyclic analogues from N-arylamides.

Dysfunction of endothelial NO system originated from homocysteine-induced aberrant methylation pattern in promoter region of DDAH2 gene.

BACKGROUND: Hyperhomocysteinemia (HHcy)-mediated dysfunction of endothelial NO system is an important mechanism for atherosclerotic pathogenesis. Dimethylarginine dimethylaminohydrolase (DDAH) is the key enzyme for degrading asymmetric dimethylarginine (ADMA), which is an endogenous inhibitor of endothelial nitric oxide (NO) synthase (eNOS). This study was designed to investigate whether the dysfunction of endothelial NO system originates from HHcy-mediated aberrant methylation modification in promotor region of DDAH2 gene. METHODS: Human umbilical vein endothelial cells (HUVECs) were cultured to the third generation and treated with homocysteine (Hcy) at different concentrations (0, 10, 30, 100, and 300 micromol/L) for 72 hours. The methylation pattern in promoter region CpG island of DDAH2 gene was analyzed by nested methylation-specific PCR (nMSP). The mRNA expression of eNOS gene and DDAH2 gene was detected by semi-quantitative RT-PCR. The activity of DDAH2 and eNOS in cells, and the concentrations of ADMA and NO in culture medium were assayed respectively. RESULTS: Mild increased concentration of Hcy (10 and 30 micromol/L) induced hypomethylation, while high concentration of Hcy (100 and 300 micromol/L) induced hypermethylation in the promoter CpG island of DDAH2 gene. The mRNA expression of DDAH2 increased in mild enhanced concentration of Hcy, and decreased in high concentration of Hcy correspondingly. The inhibition of DDAH2 activity, the increase of ADMA concentration, the reduction of eNOS activity and the decrease of NO production were all consistently relevant to the alteration of Hcy concentration. CONCLUSION: The increased concentration of Hcy induced aberrant methylation pattern in promotor region of DDAH2 gene and the successive alterations in DDAH/ADMA/NOS/NO pathway, which showed highly relevant and dose-effect relationship. The results suggested that the dysfunction of endothelial NO system induced by HHcy could be partially originated from Hcy-mediated aberrant methylation in DDAH2 gene.

Autologous transfusion with modified total hepatic vascular exclusion for extracapsular resection of giant hepatic cavernous hemangioma.

BACKGROUND: This paper was to review the effects of intraoperative autologous transfusion during modified, normal-temperature, total hepatic vascular exclusion (THVE) for extracapsular resection of giant hepatic cavernous hemangioma. METHODS: The clinical data from 28 patients, who underwent hepatic resection requiring intraoperative autologous transfusion with the cell-saver apparatus, were analyzed retrospectively. The tumors in the 28 patients involved the proximal hepatic veins and inferior vena cava. The volume of these hemangiomas ranged from 12 x 15 cm to 18 x 40 cm. All patients had varying degrees of THVE. RESULTS: The 28 patients with hemangioma received integrated resection and recovered. One patient had rupture of tumors resulting in massive hemorrhage of 6000 ml during liver resection; 4 patients had blood transfusions of 400-800 ml; the other 23 patients had no blood transfusion. Only 6 patients underwent the Pringle maneuver with resection. The other 22 patients underwent THVE during the liver resection. The interval of THVE was 5-30 minutes (mean 16 minutes). CONCLUSIONS: Intraoperative autologous transfusion during modified, normal-temperature THVE for extracapsular resection of huge hepatic cavemous hemangioma is feasible.

[Clinical investigation of the protective effects of potassium magnesium aspartate against arrhythmia and its possible anti-oxidative mechanism].

OBJECTIVE: To investigate the protective effects of potassium magnesium aspartate against oxidative stress status and lipid oxidative damage in the patients with angina and arrhythmia due to coronary artery disease, its therapeutic effect on arrhythmia and its possible mechanism. METHODS: With single blind protocol, 98 patients with angina and arrhythmia due to coronary artery disease were randomly divided into (1)Experiment group (n = 65), who received routine remedy for coronary heart disease plus potassium magnesium aspartate. (2)Control group (n = 33), who received only routine therapy for coronary heart disease without potassium magnesium aspartate. Reduced glutathione (GSH), oxidized glutathione (GSSG), malondialdehyde (MDA) and oxidized low density lipoprotein (ox-LDL) in plasma of all patients were examined before and one week after treatment, all patients with arrhythmia were equipped with Holter for continuous monitoring of cardiac rhythm. RESULTS: After one week's treatment, the GSH level in plasma of experiment group and the ratio of GSH and GSSG (GSH/GSSG) were significantly increased comparing with control group (both P<0.01), while GSSG, MDA and ox-LDL levels significantly lowered comparing with control group(all P<0.01). The premature beats diminished 86.5% in experiment group, but the decrease rate in control group was only 47.4% (P<0.01). The improvement in indexes of oxidative stress status (including GSH/GSSG, MDA and ox-LDL) and the reduction of premature beats showed close correlation with each other (all P<0.01). No adverse effects of the drug were found after one week of administration of Potassium magnesium aspartate. CONCLUSION: Potassium magnesium aspartate can strikingly improve oxidative stress status and decrease lipid oxidative damage in the patients with coronary heart disease, and the frequent premature beats were also significantly reduced by potassium magnesium aspartate. The analysis of above results reveals an intrinsic relationship between the improvement of oxidative stress status and the good therapeutic effects on frequent premature beats by potassium magnesium aspartate, which may suggest an involvement of oxidative stress in the pathogenesis of arrhythmias.

[Antagonistic effect of 3'-daidzein sulfonate sodium on prostatic hyperplasia in mice].

OBJECTIVE: To study the antagonistic effect of 3'-daidzein sulfonate sodium (DSS) on benign prostatic hyperplasia (BPH) and its possible mechanism. METHODS: Forty healthy mice were randomly divided into five groups: a normal control group without any treatment, a model group of BPH treated by subcutaneous injection of testosterone propionate, a positive control group with the BPH procedure treated by Qianliekang, a 20 mg/(kg x d) DSS group with the BPH procedure and a 40 mg/(kg x d) DSS group with the BPH procedure. After 12 days of the above treatments, the mice were sacrificed for measurement of the prostate glandular wet weight, the index of prostate gland (PI), the morphological changes of prostate gland by light microscopy and the contents of testosterone and estradiol in the serum. RESULTS: The prostate wet weight and PI decreased dose-dependently after DSS treatment for 12 days compared with the BPH model group (P < 0.05 or P < 0.01). The hyperplastic epithelioglandular papilla waned and even disappeared in the DSS treated groups under the light microscope, the epithelial cells became cubical or flat. The effect of DSS at 40 mg/(kg x d) was similar to that of the positive anti-BPH drug Qianliekang. DSS reduced the serum testosterone, estradiol contents and the T/E2 ratio (P < 0.05 or P < 0.01). CONCLUSION: DSS has significant antagonistic effect on BPH induced by testosterone propionate in mice, which may involve its regulatory action on the sex hormone balance.

[The effect of ginkgolide B on astrocyte GFAP expression after the focal cerebral ischemia].

OBJECTIVE: To observe the effect of platelet activating factor (PAF) antagonist, ginkgolide B (GB), on the expression of glial fibrillary acidic protein (GFAP) in astrocytes during focal cerebral ischemia, of which the mechanism will be explored too. METHODS: The focal cerebral ischemia of tree shrews was induced to form via photochemical reaction. Morphological changes were observed by hematoxylin-eosin staining and electron microscopic technology. The GFAP expression in astrocytes was detected by immunohistochemistry. RESULTS: Morphological changes of the brain tissue occurred after focal cerebral ischemia. Astrocytes were more swollen with the prolongation of ischemia. The GFAP expression in astrocytes in the penumbra didn't change obviously at 4 h, but increased significantly at 24 h (P < 0.01), and retained the higher level at 72 h (P < 0.01) after focal cerebral ischemia. Whereas in contralateral cortex, the GFAP expression began to increase at 72 h (P < 0.05) after focal cerebral ischemia. With giving experimental animals GB at 6 h, the GFAP expression decreased until at 24 h after focal cerebral ischemia. CONCLUSION: GFAP expression in astrocytes increases after focal cerebral ischemia. And by inhibiting the astroglial GFAP expression, the ginkgolide B exerts the cerebral protective effects.

[Gene therapy for melanoma by bifidobacterium infantis-mediated transfer of CD and UPRT genes with 5-FC in vitro].

OBJECTIVE: This study was conducted by using Bifidobacterium Infantis as a delivery system to transport suicide gene CD and UPRT to tumors in an attempt to assess the UPRT-enhanced antitumor effect of CD/5-FC. METHODS: The recombinant plasmid pGEX-UPRT was constructed and transfered into Bifidobacterium Infantis by electroporation and then identified. The synergistic antitumor effect of coexpression CD and UPRT was determined by MTT method. And the morphologic changes of B16-F10 cells were observed. RESULTS: Recombinant Bifidobacterium Infantis could express UPRT correctly. The suvival rate of cells administrated CD+ UPRT and 5-FC was significantly lower than that of control (P<0.01), and the 5-FC sensitivity (IC50 = 0.015 micromol/mL) exhibited a 8. 5-fold increase when compared with that of cells administrated CD alone (IC50 = 0.127 micromol/mL). The cells treated with CD+UPRT were remarkably damaged morphologically, and the growth of cells was significantly inhibited as compared with that of other groups. CONCLUSION: Recombinant Bifidobacterium Infantis with UPRT gene can significantly enhance the killing effect of CD/5-FC suicide gene system on melanoma B16-F10 cells of mice.

[Effect of decreased GSH on sensitivity of breast cancer cells to ADM].

OBJECTIVE: Glutathione(GSH) maintains an optimum cellular redox potential. Elevated levels of GSH render some types of cancer cells resistant against anti-cancer drugs. The aim of this study was to determine the effect of a thiol-depleting agent, diethylmaleate (DEM), on the sensitivity of human breast cancer cells to ADM. METHODS: The ADM-resistant human breast cancer MCF-7/ADM cell lines and ADM-sensitive MCF-7/S cell lines were treated by thiol-depleting agent DEM for 3 h respectively. The changes of sensitivity to ADM were then measured by MTT assay. The intracellular GSH contents were examined by fluorescent-spectrophotometry and the correlation between the changes of sensitivity to ADM and the intracellular GSH content was analyzed. RESULTS: Treatment of MCF-7/ADM and MCF-7/S cells by 0.1 micromol/L DEM for 3 h decreased 37.4% and 29.7% of the intracellular GSH content respectively (P < 0.01). ADM also decreased intracellular GSH content in a ADM-concentration-dependent manner. The combined use of DEM and ADM depleted the intracellular GSH content in both cells significantly more than the sum of single use of ADM and DEM alone. The sensitivity of both cells to ADM increased with the decline of intracellular GSH content. CONCLUSION: The depletion effect of DEM on the intracellular GSH could be enhanced by ADM and such depletion may be involved in the changes of the sensitivity of MCF/7 cells to ADM.

[Effects of peroxisome proliferators activated receptors on caveolin-1 expression in foam cells].

OBJECTIVE: To study the effect of peroxisome proliferators activated receptors (PPAR) alpha, gamma ligand on ATP-binding cassette transporter A1 (ABCA1) and caveolin-1 expressions and cholesterol, ox-LDL contents in human monocyte derived foam cells. METHOD: Malondialdehyde (MDA) was measured by TBARS method, ox-LDL detected by ELISA method, cholesterol measured by fluorescence spectrophotometric method, ABCA1, caveolin-1 mRNA and protein expressions determined by RT-PCR and Western blot, in human monocytes, foam cells [human monocyte-derived macrophage induced by myristate acetate (PMA) further treated with 50 mg/L ox-LDL for 24 h], foam cells plus 10 micromol/L pioglitazone for 48 h, foam cells plus 5 micromol/L clofibrate for 48 h. RESULT: The intracellular total cholesterol (TC), free cholesterol (FC), cholesteryl ester (CE), ox-LDL and lipid peroxide were significantly increased and the membrane expressions of ABCA1, caveolin-1 were down-regulated in foam cells compared to monocytes (all P < 0.05) and these changes were significantly attenuated by cotreatment with PPARalpha, gamma ligand. CONCLUSION: The anti-atherosclerosis effects of PPARalpha, gamma ligand are related to reducing cholesterol contents and up-regulating ABCA1, caveolin-1 expressions in foam cells.

[Experimental study of anti-atherosclerosis potency by lycium seed oil and its possible mechanism].

OBJECTIVE: To study the effect of anti-atherosclerosis of Lycium Seed Oil (Lso) and its possible mechanism. METHODS: The rabbit atherosclerosis model was established by high fat diet, and the TC, TG, LDL-C, HDL-C levels in plasma were examined dynamically. The SOD, GSH-PX, T-AOC activities and the MDA levels in serum were monitored after 8 week's high fat diet. Aorta samples were observed for atherosclerotic extent, and NF-kappaB, TNF-alpha were assessed by immuno-histochemical method. The lovastatin was set up as a positive control. RESULTS: contents of HDL-C obviously increased in Plasma of low and high dosage groups and TC, TG, LDL-C levels significantly decreased compared with control group. The SOD, GSH-PX, T-AOC activities up-regulated while the NF-kappaB, MDA and NF-alpha levels decreased in Lycium Seed Oil groups compared with control group. Aortic atherosclerotic extent and area in low dosage and high dosage LSO groups were absolutely smaller than that in high fat diet group. The anti-atherosclerosis potency of Lycium Seed Oil was similar with that of lovastatin. CONCLUSION: Lycium Seed Oil has potent anti-atherosclerosis effects and its anti-atherosclerosis potency was similar with The lovastatin. The possible mechanism involve the decreasing of plasma lipids, anti-peroxidation, inhibiting the activation of NF-kappaB and down-regulating the inflammation cytokines of TNF-alpha.