Objective To obtain antibiotic-resistant mutants producing metabolites with antitumor activity from wild-type actinomycete strains without antitumor activity. Methods An actinomycete strain L35-1 was used as an initial strain for obtaining antibiotic-resistant mutants, which is a marine-derived wild-type strain without antitumor activity with an inhibition rate of 2.8% at the 1000 μg/ml of high sample concentration on K562 cells. The antibiotic-resistant mutants both from auto-mutagenesis and chemical mutagen-induced mutagenesis were selected by single colony isolation on antibiotic-containing plates according to the method for obtaining drug-resistant mutants in ribosome engineering. The antitumor activity was assayed by the MTT method using K562 cells for the mutants with aqueous acetone extracts of the whole broth of their fermentation.Results A total of 114 neomycin-resistant (ner) and 68 streptomycin-resistant (str) mutants, all from auto-mutagenesis, was obtained on drug-containing plates. Among them, the 7 ner and 3 str mutants appeared to be bioactive with an inhibition rate above 20% at the 100 μg/ml sample concentration on K562 cells. On the other hand, 41 str and 32 ner mutants from DES-induced mutagenesis and 46 ner mutants from NTG-induced mutagenesis were obtained by mutagen-induced mutation coupled with the single colony isolation on antibiotic-containing plates, among which, one str mutant from DES-induced mutagenesis and one ner mutant from NTG-induced mutagenesis were bioactive with an inhibition rate over 20% at the 100 μg/ml sample concentration on K562 cells. Conclusions The present result has revealed that the wild-type actinomycete strains without bioactivity might become a great source initial strains to obtain bioactive mutants by drug-resistant mutation technique.

AlM: To investigate the relationship between the anterior ischemic optic neuropathy ( AlON ) and the carotid artery change using doppler ultrasound.METHODS:Fifty-four cases of AlON patients and 54 cases of healthy control were observed, atherosclerotic spots were detected by the application of color ultrasound.RESULTS:ln AlON group of 54 patients, 38 cases appeared carotid atherosclerosis, accounting for 70%. The number of cases with hard plaque, soft plaque and mixed plaques were 18, 13, and 7 respectively, accounting for 33%, 24% and 13%. ln the control group, 20 cases were detected atherosclerotic change, accounting for 37%. And the number of cases with hard plaque, soft plaque and mixed plaques were 12, 5 and 3 respectively, accounting for 22%, 9%, 6%. Significant stenosis and velocity change were showed in neither AlON group nor control group. Compared with the control group, AlON group had more cases of atherosclerotic plaque, the difference was statistically significant (χ2=12. 836, P=0. 005)CONCLUSlON: The incidence of AlON is correlated with carotid atherosclerosis, and carotid ultrasonography is significantly valuable for AlON etiology and diagnosis.

Objective To explore the influence of hemodialysis(HD) and henodiafiltation on serum ?_2-microglobulin(?_2-MG) of children with acute renal failure and contrast research on declining rate of serum ?_2-MG before and after(children′s) HD and HDF.(Met-)hods By Branc Dialog HD machine of double-pump,HDF for 18 times and HD for 20 times were given to children with acute renal(fai-)lure.The serum ?_2-MG were observed before and after HD and HDF.Results A great difference was observed in level of serum ?_2-MG between before and after HD and HDF.In HDF group,there was significant difference in level of serum ?_2-MG before and after HDF(P

OBJECTIVETo investigate effects of zearalenone (ZEA) on the proliferation of SK-N-SH human neuroblastoma cells in vitro and its possible mechanism.

METHODSSK-N-SH cells were cultured in estrogen-free improved minimum essential medium and divided into 5 groups based on different treatments: group 1, without treatment; group 2, treated with 17beta-estradiol (E(2)); group 3, treated with ZEA; group 4, treated with both E(2) and ICI 182780; group 5, treated with both ZEA and ICI 182780. Absorbance value (AV) was determined at the time point of 0, 24, 48 and 72 hours, and DNA proliferation index (PI) at 72 hours. Flow cytometer, terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) were employed to monitor cell apoptosis.

RESULTSAt 24, 48 and 72 hours, the AV of group 3 were 1.39, 1.32, and 1.22 times to those of group 1, respectively. PI in group 3 was 1.43 times of that in group 1 at 72 hours. The results of group 2 were similar to those in group 3. At the same time, the growth of cells was inhibited by ICI 182780 despite the presence of E(2) and ZEA. Apoptosis cells were abundant in group 1 and ICI 182780 groups, but little in E(2) and ZEA groups.

CONCLUSIONZEA might promote the proliferation of SK-N-SH cells to a level similar to that of E(2), which might probably be brought about via estrogen receptor pathways and depressing apoptosis.

Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Humans ; Neuroblastoma ; Receptors, Estrogen ; antagonists & inhibitors ; Zearalenone ; toxicity

Objective To discuss the application of the hemodialysis and filtration(HDF) in children with acute renal failure(ARF).Methods By Branc Dialog hemodialysis machine of double-pump,HDF for 19 times and hemodialysis(HD) for 21 times were given to the children with ARF.The serum blood urea nitrogen(BUN),creatinine(Cr),uric acid(UA),?_2 microglobulin(?_2-MG) and the rate of the heart,blood pressure,temperature were observed before and after HD and HDF.Results The clearance rates of BUN,Cr,UA,(?_2-)MG of HDF were markedly higher than those of HD(P

Objective To study the mechanism of marcosomia by investigating insulin-like growth factor 2(IGF_2)imprinting status,expression level and the promoter usage in the placenta of macrosomia. Methods We selected heterozygous cases for Apa Ⅰ polymorphism in exon 9 of IGF_2 gene and then analyzed its imprinting status in 168 placentas of macrosomia and normal pregnancies.IGF_2 transcription levels and promoter usages in macrosomic and normal placenta were evaluated by using semi-quantitative RT- PCR assay.Results Thirty specimens of macrosomic placenta and 30 of normal placenta were identified as heterozygous for IGF_2.All of the heterozygous specimens showed maintenance of imprinting.The expression of placental IGF_2 mRNA(2.2?1.2)was significantly higher in macrosomia than that of normal weight group (1.6?0.6,P 0.05).Conclusion It is possible that over expression of IGF_2 in placenta contributes to macrosomia while the promoter usage and imprinting status are not associated with macrosomia.

OBJECTIVETo explore the effects of p38MAPK inhibitor SB203580 (SB) on the occurrence of acute GVHD and intestine damage after allogeneic hematopoietic stem cell transplantation (allo-HSCT) in mice.

METHODSSixty BALB/c mice, as recipients, were randomized to control group, irradiation group, model group and intervention group. C57BL/6 mice, as donors, were raised to prepare the bone marrow cells (BMCs) and spleen cells (SCs), which were injected into irradiated recipients mice by tail vein. Except control group, other groups accepted 7.5Gy total body irradiation. Model group and intervention group were infused with BMCs 5×10⁶ and SCs 5×10⁵ by less than 4 h after irradiation. SB was injected into intervention group by intraperitoneally, but only DMSO for model group. The general status and survival rate of each group were evaluated. The expression of p-p38MAPK, Fas and FasL in intestine were determined by RT-PCR, Western blot and immunohistochemistry (IHC).

RESULTSThe weight changes of intervention group (13.00±0.50)% was significantly lighter than that of model group (25.00±0.75)% (P<0.05). The clinical score of acute GVHD in the intervention group (3.33±0.82) was significantly lower than that of model group (6.33±1.36) (P<0.05). The expression levels of p-p38MAPK, Fas and FasL in small intestine of intervention group (1.43±0.02, 0.81±0.03, 0.97±0.03) were lower than those of model group (1.76±0.05, 1.52±0.04, 1.48±0.04).

CONCLUSIONSB inhibited the activation of p38MAPK and Fas/ FasL signal pathway and alleviated the apoptosis of small intestine. And SB could relieve small intestine damages induced by allogeneic T lymphocytes.

Animals ; Apoptosis ; drug effects ; Bone Marrow Transplantation ; adverse effects ; Fas Ligand Protein ; metabolism ; Graft vs Host Disease ; metabolism ; pathology ; Imidazoles ; pharmacology ; Intestines ; drug effects ; pathology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Pyridines ; pharmacology ; Signal Transduction ; drug effects ; Transplantation, Homologous ; fas Receptor ; metabolism ; p38 Mitogen-Activated Protein Kinases ; antagonists & inhibitors ; metabolism

OBJECTIVETo evaluate whether or not administration of folic acid and resveratrol have preventive effects on cleft palate formation as well as the comparison of the two drugs' s effects.

METHODSPregnant mice were randomly divided into 9 groups, with 8 mice in each group. The TCDD group mice were dosed with TCDD 28 microg/kg body weight on gestation day 10 (GD 10) animals in folic acid group were respectively dosed with folic acid 15, 10, 5 mg/kg and TCDD 28 microg/kg; resveratrol treated mice were divided into 3 groups: resveratrol 50 mg/kg were orally administered for 6 consecutive days, from gestational day GD 8 to GD13 in resveratrol (GD8-13 ) group; resveratrol 50 mg/kg were orally administered for 6 consecutive days, from gestational day GD 8 to GD13, followed hy an oral administered with TCDD on GD10 in resveratrol (GD8-13) + TCDD group; resveratrol 50mg/kg and TCDD 28 microg/kg were used by gavage administration at GD10 in resveratrol (GD10) + TCDD group. Control mice were treated with the same volume of water for 6 consecutive days from GD8 to GD13 and were given a single dose of corn oil on GD10. The pregnant mice weight and embryos, the number of live, cleft palate, dead and resorption fetal mice were recorded on GD 17.5. The coronal sections of the fetal mice heads were prepared at GD 17.5 and observed by microscopy.

RESULTSTotal frequency of clefts was 92.86% in TCDD group, 84.00% (15 mg), 73.08% (10 mg), 84.00% (5 mg) in folic acid + TCDD groups, 0% in resveratrol (GD10) group, 74.51% (GD10), 57.78% (GD8-13) in resveratrol + TCDD groups. The frequency of cleft was 0% in the control group. Compared with the control and the TCDD groups, there were significant differences in the number of live, dead and resorption fetal mice in TCCD + resveratrol (GD8-13) group (P < 0.05). No significant differences in embryonic weight, live fetuses weight, the number of live, dead and resorption fetal mice were found in the other groups (P > 0.05).

CONCLUSIONTest dose of folic acid and resveratrol both had certain antagonistic effect on cleft palate in mice induced by TCDD, with folic acid 10 mg/kg, resveratrol 50 mg/kg GD8-13 doses having stronger antagonistic action. Effects of both the two drugs have no significant difference, but resveratrol (50 mg/kg, GD8-13) significantly affects the fetal mice's growth and development under TCDD exposure in utero.

Abnormalities, Drug-Induced ; prevention & control ; Animals ; Cleft Palate ; chemically induced ; prevention & control ; Female ; Fetus ; Folic Acid ; administration & dosage ; pharmacology ; Humans ; Mice ; Mice, Inbred C57BL ; Polychlorinated Dibenzodioxins ; antagonists & inhibitors ; Pregnancy ; Random Allocation ; Stilbenes ; administration & dosage ; pharmacology ; Teratogens

To investigate the effects of lanosterol (1), inotodiol (2) and trametenolic acid (3) from Inonotus obliquus against oxidative damage induced by CCl4 in mice, 1, 2 and 3 (20, 10 and 5 mg x kg(-1)) were respectively administered to mice, once a day for 3 days. Then the mice were induced to oxidative damage by CCl4 on the third day 30 min after the administration. The activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-PX) and the content of malondialdehyde (MDA) and reductive glutathione (GSH) in serum and liver homogenate were determined. And the levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) and interleukin-6 (IL-6) concentration in serum were detected. The results showed that treatment with compound 1, 2 and 3 could significantly increase the activities of SOD, CAT and GSH-PX in serum and liver homogenate. Furthermore, the content of GSH in serum and liver homogenate increased and MDA content decreased markedly. In addition, compound 1, 2 and 3 could significantly inhibit the activities of ALT and AST in serum, and decrease the IL-6 concentration in serum remarkably. So, compound 1, 2 and 3 can protect mice against oxidative stress injury induced by CCl4. Furthermore, compound 1, 2 and 3 can protect cells from damage through inhibition on ALT, AST and the expression of IL-6.
Alanine Transaminase ; blood ; Animals ; Aspartate Aminotransferases ; blood ; Carbon Tetrachloride ; Catalase ; blood ; metabolism ; Female ; Glutathione ; blood ; metabolism ; Glutathione Peroxidase ; blood ; metabolism ; Interleukin-6 ; blood ; Lanosterol ; analogs & derivatives ; isolation & purification ; pharmacology ; Liver ; metabolism ; Male ; Malondialdehyde ; blood ; metabolism ; Mice ; Oxidative Stress ; drug effects ; Polyporaceae ; chemistry ; Protective Agents ; isolation & purification ; pharmacology ; Random Allocation ; Superoxide Dismutase ; blood ; metabolism ; Triterpenes ; isolation & purification ; pharmacology

OBJECTIVETo study the effect of testosterone propionate (TP) on the distribution pattern of calcitonin gene-related peptide (CGRP) in two types of motoneuron (Mn) pools in rats.

METHODThe double labeling of cholera toxin B subunit coupled with colloidal gold (CB-Au) retrograde identification combining with immunocytochemistry was mainly used to reveal the distribution pattern of CGRP-like immunoreactivity (CGRP-LI) and its changes in the motoneuron pools labeled by CB-Au.

RESULTTP injected intramuscularly 28 days later significantly decreased CGRP expression in Mn pool innervating extensor digitorum longus (EDL, fast-twitch), comparing with corresponding control and castration group respectively (P < 0.001), while no significant effect on Mn pools innervating soleus (SOL, slow-twitch, P > 0.05) was observed.

CONCLUSIONEDL-Mn pool is more sensitive to testosterone propionate than SOL-Mn pool in regulating CGRP expression.

Animals ; Calcitonin Gene-Related Peptide ; drug effects ; metabolism ; Male ; Motor Neurons ; drug effects ; metabolism ; Muscle Fibers, Fast-Twitch ; cytology ; drug effects ; Muscle Fibers, Slow-Twitch ; cytology ; drug effects ; Rats ; Rats, Wistar ; Testosterone Propionate ; pharmacology