Experimental St■■■es on the Mutagenicity of Hydroxypiperaquine Phosphate

The mutagenicity of hydroxypiperaquine phosphate (HPQP) was detected by a battery of assays including Amestest, chromosomal aberration test in vitro, micronucleus testand dominant lethal test The results obtained have shownthe absence of mutagenicity of HPQP up to the inhibitoryconcentration to Salmonella typhimurium TA97a. TA98.TA100 and TA102 with or without S-9 mix. The incidence     

Studies on the Mutagenicity of Hair Dyes Made in China

A total of 13 commercial hair dye products made in China were tested for mutagenicity in 2 short-term bioassays, the histidine-requiring mutants of Sahnonella typhhnurium (strains TA98 and TA100) and the micronucleus test with mouse bone-marrow polychromatic erythrocyte cells in vivo. The results showed that the 13 hair dyes were not mutagenic in strains TA98 and TA100 with and without S-9. In the micronucleus test, no mutagenic effect was observed.     

Study on the Mutagenicity of Diesel Exhaust Particles

The mutagenicity of diesel exhaust particles(DEP)was studied by using Salmonella typhimurium strains TA98 and TA100 in vitro and mice micronucleus in vivo test.DEP from six kinds of medium and heavy-duty diesel vehicles,which were made in China and imported,were tested.The vehicles wer eoperated under free accelerating condition.The results showed that the EP contained mutagenic activity.An increase in the number of the Salmonella TA98 was observed in the presence and especially in the absence of S9 Mmix.Positive results were also obtained from mice micronucleus assay.The frequency of mice bone marrow micronucleated polychromatic erythrocytes(M PCE)was increased and it showed a definite dose-response relationship.Comparing the different types of the vehicles,we found that the mutagenicity of DEP from domestic made vehicles was stronger than that from the imported ones.     

The Mutagenicity of the Lyophilized Agkistrodon Haly （pallas） Venom

Objective To investigate the mutagenicity of Methods The mutagenicity of the Lyophilized Agkistrodon the Lyophilized Agkistrodon Haly （pallas） Venom. Haly （pallas） Venom was studied by the micronucleus test, the Ames test and the CHL test. Results No significant differences in the micronuclear frequences were found in any of the testing groups compared with negative control in mice （p〉0.05）; The mutagenic activity of the Lyophilized Agkistrodon Haly （pallas） Venom was not observed in TA97, TA98, TA100 and TA102 with and without S9 activation under the doses used; In CHL test, the effect of chromosomal damage of the Lyophilized Agkistrodon Haly （pallas） Venom was not found either in CHL cells. Conclusion The Lyophilized Agkistrodon Haly （pallas） Venom has no mutagenicity, and it might be useful in future.     

A Study on Genotoxicity of Cooking Fume from Rapeseed Oil

The present article reports the genotoxic potential of rapeseed oil cooking fume investigated by a battery of short-term tests (Ames test, SCE/V79 in vitro and mice micronucleus in vivo test). The results showed that the cooking fume contained mutagenic activity. In the presence of S9 mix, an increase in the number of the Salmonella TA98 was observed at doses ranging from 1.0 to 5.0 mg/plate, and the SCE frequencies of V79 cell were markedly raised at doses ranging from 0.05 to 0.5 mg·ml~(-1). The positive result was also obtained in mice micronucleus assay, the mice had inhaled the cooking fume a week earlier. The frequency of mice bone marrow MN-PCE was increased and it showed a remarkable time-dose-response relationship during the 4 weeks exposure. The results suggested that this cooking fume exposure may be a risk factor of lung cancer in Chinese women.     

Effect of behavior training on learning,memory and the expression of NR2B,GluR1 in hippocampus of rats offspring with fetal growth restriction

Objective： To study effects of behavior training on learning, memory and the expression of NR2B, GluR1 in hippocampus of rat＇ s offspring with fetal growth restriction（FGR）. Methods： The rat model of FGR was established by passive smoking method. The rats offspring were divided into the FGR group and the control group, then randomly divided into the trained and untrained group, respectively. Morris water maze test was proceeded on postnatal month（PM2/4） as a behavior training method, then the learning-memory of rats was detected through dark-avoidance and step-down tests. The expressions of NR2B and GluR1 subunits in hippocampal CA1 and CA3 areas were detected by immunohistochemical method. Results： In the dark-avoidance and step-down tests, the performance record of rats with FGR was worse than that of control rats, and the behavior-trained rats was better than the untrained rats, when the FGR model and training factors were analyzed singly. The model factor and training factor had significant interaction（P 〈 0.05）. The expressions of NR2B and GluR1 subunits in hippocampal CA1 and CA3 areas of rats with FGR reduced. In contrast, the expressions of GluR1 and NR2B subunits in CA1 area of behavior-trained rats increased, when the FGR model and training factors were analyzed singly. Conclusion： These findings suggested that the effect of behavior training on the expressions of NR2B and GluR1 subunits in CA1 area should be the mechanistic basis for the training-induced improvement in learning-memory abilities.     

Effect of behavior training on learning,memory and the expression of NR2B,GluR1 in hippocampus of rats offspring with fetal growth restriction

Objective:To study effects of behavior training on learning,memory and the expression of NR2B,GluR1 in hippocampus of rat's offspring with fetal growth restriction(FGR).Methods:The rat model of FGR was established by passive smoking method.The rats offspring were divided into the FGR group and the control group,then randomly divided into the trained and untrained group,respectively. Morris water maze test was proceeded on postnatal month(PM2/4)as a behavior training method,then the learning-memory of rats was detected through dark-avoidance and step-down tests.The expressions of NR2B and GluR1 subunits in hippocampal CA1 and CA3 areas were detected by immunohistochemical method.Results:In the dark-avoidance and step-down tests,the performance record of rats with FGR was worse than that of control rats,and the behavior-trained rats was better than the untrained rats,when the FGR model and training factors were analyzed singly.The model factor and training factor had significant interaction(P<0.05).The expressions of NR2B and GluR1 subunits in hippocampal CA1 and CA3 areas of rats with FGR reduced.In contrast,the expressions of GluR1 and NR2B subunits in CA1 area of behavior-trained rats increased,when the FGR model and training factors were analyzed singly. Conclusion:These findings suggested that the effect of behavior training on the expressions of NR2B and GluR1 subunits in CA1 area should be the mechanistic basis for the training-induced improvement in learning-memory abilities.     

Effects of isoflurane on ICAM-1 expression and neutrophils infiltration in rats with liver ischemia and reperfusion injury

Objective： To establish a rat model of warm partial hepatic ischemia-reperfusion （IR）, and investigate the protective and anti-inflammatory effects of isoflurane on warm hepatic ischemia-reperfusion injury （IRI） in rats. Methods： Thirty-two female Sprague-Dawley rats were divided equally into 4 groups （n-8）： PB-Sham group in which the rats were anesthetized by intraperitoneal injection of pentobarbital sodium （1.0%, 40 mg/kg, PB） and received a sham operation without occlusion of liver blood flow; PB-IR group whose rats underwent partial hepatic IR after anesthesia; Iso-Sham group in which inhalation of 1.0 MAC isoflurane and sham operation was performed; Iso-IR group in which 1.0 MAC isoflurane was inhaled for 4 h and IR was performed. Rat model of warm partial hepatic IR was established by clamping the hepatic arteries and hilar vessels distributing to the left and median lobes to induce partial hepatic ischemia （70%） for 60 rain followed by reperfusion for 3 h. The rats were killed 3 h after declamping, and specimens of liver tissue and blood were obtained. The serum ALT and AST were detected as liver damage markers. Viability of myeloperoxidase （MPO） in liver was measured. The protein level of ICAM-1 in the liver was detected by immunohistochemistry and Western blotting. Results： Rats treated with 1.0 MAC isoflurane during warm partial （70%） hepatic ischemia 60 rain and 3 h reperfusion had significantly lower serum ALT and AST compared with rats anesthetized with pentobarbital sodium subjected to hepatic IRI. The expression of ICAM-1 in hepatic tissue was significantly increased by hepatic IRI after pentobarbital sodium anesthesia. Isoflurane significantly inhibited protein expression of ICAM-1 in hepatic IR injury compared with pentobarbital sodium anesthesia. Viability of liver MPO was significantly increased by hepatic IRI after pentobarbital sodium anesthesia; Isoflurane can significantly inhibit MPO alteration in rat liver ischemia-reperfusion injury compared with rats anesthetized with pentobarbital sodium. Conclusion： Isoflurane anesthesia can attenuate liver IR injury in rats that maybe by inhibiting ICAM-I expression and reducing the infiltration of neutrophils.     

Influence of phenobarbital and carbon tetrachloride on the modulation of tissue retention profile of hexachlorocyclohexane in rats

The study was designed to investigate the role of hepatic metabolic activity on body burden of HCH residue. Male albino rats were orally administered 0, 5, and 10 mg/kg HCH for 90 days, followed by either sodium phenobarbital or carbon tetrachloride treatment for 0, 15 and 30 days after withdrawal of their respective HCH administration. The liver weight was significantly increased at 30 days after the administration of phenobarbital and carbon tetrachloride in both 5 mg and 10 mg/kg HCH withdrawal groups when compared to control. HCH residue was maximun in fat followed by adrenal>thymus>liver>kidney>spleen>testes>brain>plasma. Carbon tetrachloride caused an accumul-ation of HCH residues in the liver 15 and 30 days after administration of both doses of HCH. Phenobarbital did not show significant variation in HCH residues in hepatic tissue. Phenobarbital treatment caused significant induction of hepatic RED, APD, AHH, GST and QR activities. Significant decreases in activities were observed by carbon tetrachloride when compared to animals treated with HCH alone. The overall results clearly suggest the role of P450 protein on the body burden of HCH residues.     

Scanning electron microscopic observation: three-dimensional architecture of the collagen in hepatic fibrosis rats

Background In the process of hepatic fibrosis, the accumulation of collagen fibers is strongly related to the hepatic function. The aim of this study was to investigate the three-dimensional architecture of the collagen network in the liver of rats with hepatic fibrosis. Methods Healthy adult male Wistar rats （n=32） were randomly divided into a control group （n=16） and a hepatic fibrosis group （n=16）. In the control group, the rats were treated with peanut oil while the rats in hepatic fibrosis group were treated for 10 weeks with 60% CCI4 diluted in peanut oil. The quantity of collagen fibers was detected by Western blotting; distribution of the collagen was detected by sirius red staining and polarized microscope; the three-dimensional architecture of collagen in the liver was observed under the scanning electron microscope after fixed tissues were treated with cell-maceration using NaOH. Statistical analysis was performed using the u test. Results The quantity of collagen fibers increased significantly in the hepatic fibrosis group. With the aggravation of hepatic fibrosis, collagen fibers gradually accumulated. They interlaced the reticulation compartment and formed a round or ellipse liver tissue conglomeration like a grape framework that was disparate and wrapped up the normal liver Iobule. The deposition of collagen fibers was obvious in adjacent hepatic parenchyma, especially around the portal tracts. Conclusion Our experiment showed the collagen proliferation and displays clearly the three-dimensional architecture of collagen fibers in rat liver with hepatic fibrosis by scanning electron microscope. It can provide a morphological foundation for the mechanisms of changed haemodynamics and portal hypertension in hepatic fibrosis.     

Effects of hepatotrophic factors on the liver after portacaval shunt in rats with portal hypertension

Background Portacaval shunt （PCS） prevent hepatotrophic factors from flowing into the liver, but they enter directly the systemic circulation and worsen liver injury. This study was designed to investigate the effects of hepatotrophic factors through the portal vein on the liver in rats with portal hypertension after portacaval shunt. Methods Intrahepatic portal hypertension （IHPH） was induced by intragastric administration of carbon tetrachloride, and end-to-side PCS was performed. Eight normal rats served as controls, and eight rats with IHPH served as IHPH model （IHPH group）. Another 32 rats with IHPH-PCS were randomly subdivided into 4 groups： normal saline （NS） given to 8 rats, hepatocyte growth factor （HGF） 8, insulin （INS） 8, hepatocyte growth factor and insulin （HGF＋INS） 8. Hepatotrophic factors were infused into the portal vein through an intravenous catheter. Portal venous pressure （PVP） was measured. The levels of serum alanine aminotransferase （ALT） and aspartate aminotransferase （AST） were tested biochemically and those of hyaluronic acid （HA） and laminin （LN） were measured by radioimmunoassay. Hepatic fibrosis was assessed histologically and the expression of collagens type I and In were detected immunohistochemically. Ultrastructural change of hepatocytes and the number of mitochondria were observed under an electron microscope. The data were compared between groups and subgroups by Student-Newman-Keuls procedure with SPSS 10.0. Results PVP was significantly higher in the IHPH rats than in the control rats （P〈0.05）. The levels of serum ALT, AST, HA, and LN, hepatic fibrosis score, the amount of collagen deposition, collagens type I and III increased more significantly in the IHPH group than in the control rats （P〈0.05）. The number of mitochondria decreased more significantly in the IHPH rats than in the control rats （P〈0.05）. The levels of serum ALT, AST, HA and LN as well as hepatic fibrosis score, the amount of collagen deposition, and the amount of collagens type I and M in the HGF and HGF＋INS rats were significantly lower than those in the NS rats （P〈0.05）. The damage to hepatocyte ultrastructure was markedly alleviated and the number of mitochondria was increased more significantly in the HGF and HGF＋INS rats than in the NS rats under an electron microscope. Conclusions Perfusion of exogenous hepatotrophic factors through the portal vein can alleviate liver injury, minimize the damage to the ultrastructure of hepatocyte, protect liver function, and lessen hepatic fibrosis in rats with portal hypertension after PCS.     

Impaired upregulation of keratinocyte growth factor in injured lungs induced by Pseudomonas aeruginosa in immunosuppressed rats

Background The number of immunosupressed patients has increased in the past decades. Among them Pseudomonas aeruginosa （P. aeruginosa） is one of the leading bacteria for pneumonia that are associated with poor prognosis. However, the pathogenesis of P. aeruginosa pneumonia in immunosupressed patients is not understood completely. Previous reports showed keratinocyte growth factor （KGF） is associated with lung injury in immunocompetent hosts. In this study, we investigated the different reactions of lung injury, lung pathology and KGF expressions in P aeruginosa pneumonia between immunosuppressed and immunocompetent rats. Methods Immunosuppression of male rats was induced by injecting immunosuppressive subcutaneously. Pneumonia was established by instilling P aeruginous tracheally. The immunocompetent rats were the control group. Survival rate, lung histopathology, pulmonary permeability and oedema, KGF mRNA and protein expressions in lungs of both groups were investigated. Results The survival rate of immunosuppressed group was lower than that of immunocompetent group （33.3% vs 83.3%）. After exposure to bacteria, pulmonary permeability and wet/dry ratio in immunosuppressed group were higher than those in immunocompetent group. Pulmonary congestion and haemorrhage were more intensive in immunosuppressed group compared to immunocompetent group. Apoptosis and necrosis were also observed in infected lungs of immunosuppressed rats. Although we detected KGF expressions in lungs of both groups after infection, the expressions of KGF protein and mRNA gene in immunosuppressed group were much lower than in immunocompetent group. Conclusions Compared with immunocompetent group, there was more intensive lung injury in immunosuppressed group. Severe lung injury may contribute to the poor prognosis of pneumonia. KGF expressions of pneumonia in immunosuppressed rats were less than those in immunocompetent ones.     

Antioxidant and Antigenotoxic Activity of Bioactive Extracts from Corn Tassel简

This study is designed to evaluate antioxidant and antigenotoxic activities of corn tassel extracts（CTTs）. The major bioactive components of CTTs include flavonoid, saponin and polysaccharide. The antioxidant properties of the three bioactive components of CTTs were investigated by Ferric Reducing Antioxidant Property（FRAP） and 1, 1-diphenyl-2-picrylhydrazyl（DPPH） assays. The activities of the extracts were determined by assessing the inhibition of mutagenicity of the direct-acting mutagen fenaminosulf, sodium azide, and indirect-acting mutagen 2-aminofluorene using the Ames test（strains TA98 and TA100）. The results showed that the extraction rates of flavonoid, saponin, and polysaccharide from the dried corn tassels were 1.67%, 2.41% and 4.76% respectively. DPPH and FRAP assay strongly demonstrated that CTTs had antioxidant properties. CTTs at doses of 625, 1250 and 2500 μg per plate reduced 2-aminofluorene mutagenicity by 12.52%, 28.76% and 36.49% in Salmonella typhimurium TA98 strain assay respectively and by 10.98%, 25.27% and 37.83%, at the same doses in Salmonella typhimurium TA100 assay system, respectively. 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide（MTT） assay showed that the different concentrations of CTTs inhibited the proliferation of MGC80-3 cells in a dose-dependent manner（P0.01）. It is concluded that these integrated approaches to antioxidant and antigenotoxicity assessment may be useful to study corn tassel as a natural herbal material.     

Propofol can Protect Against the Impairment of Learning-memory Induced by Electroconvulsive Shock via Tau Protein Hyperphosphorylation in Depressed Rats

Objective To explore the possible neurophysiologic mechanisms of propofol and N-methyl-D-aspartate (NMDA) receptor antagonist against learning-memory impairment of depressed rats without olfactory bulbs.
Methods Models of depressed rats without olfactory bulbs were established. For the factorial design in analysis of variance, two intervention factors were included: electroconvulsive shock groups (with and without a course of electroconvulsive shock) and drug intervention groups [intraperotoneal (ip) injection of saline, NMDA receptor antagonist MK-801 and propofol. A total of 60 adult depressed rats without olfactory bulbs were randomly divided into 6 experimental groups (n=10 per group):ip injection of 5 ml saline;ip injection of 5 ml of 10 mg/kg MK-801;ip injection of 5 ml of 10 mg/kg MK-801 and a course of electroconvulsive shock;ip injection of 5 ml of 200 mg/kg propofol;ip injection of 5 ml of 200 mg/kg propofol and a course of electroconvulsive shock;and ip injection of 5 ml saline and a course of electroconvulsive shock. The learning-memory abilities of the rats was evaluated by the Morris water maze test. The content of glutamic acid in the hippocampus was detected by high-performance liquid chromatography. The expressions of p-AT8Ser202 in the hippocampus were determined by Western blot analysis.
Results Propofol, MK-801 or electroconvulsive shock alone induced learning-memory impairment in depressed rats, as proven by extended evasive latency time and shortened space probe time. Glutamic acid content in the hippocampus of depressed rats was significantly up-regulated by electroconvulsive shock and down-regulated by propofol, but MK-801 had no significant effect on glutamic acid content. Levels of phosphorylated Tau protein p-AT8Ser202 in the hippocampus was up-regulated by electroconvulsive shock but was reduced by propofol and MK-801 alone. Propofol prevented learning-memory impairment and reduced glutamic acid content and p-AT8Ser202 levels induced by electroconvulsive shock.
Conclusion Electroconvulsive shock might reduce learning-memory impairment caused by protein Tau hyperphosphorylation in depressed rats by down-regulating glutamate content.     

Effects of Puerarin on Pulmonary Vascular Remodeling and Protein Kinase C-α in Chronic Cigarette Smoke Exposure Smoke-exposed Rats

In order to investigate the effects of puerarin on pulmonary vascular remodeling and protein kinase C-α (PKC-α) in chronic exposure smoke rats, 54 male Wistar rats were randomly di- vided into 7 groups: control group (C group), smoke exposure groups (S4w group, Saw group), puer- arin groups (P4w group, P8w group), propylene glycol control groups (PC4w group,PC8w group). Rats were exposed to cigarette smoke or air for 4 to 8 weeks. Rats in puerarin groups also received puer- arin. To evaluate vascular remodeling, alpha-smooth muscle actin (α-SM-actin) staining was used to count the percentage of completely muscularised vessels to intraacinar pulmonary arteries (CMA/IAPA) which was determined by morphometric analysis of histological sections. Pulmonary artery smooth muscle cell (PASMC) apoptosis was detected by in situ end labeling technique (TUNEL), and proliferation by proliferating cell nuclear antigen (PCNA) staining. Reverse transcrip- tion-polymerase chain reaction (RT-PCR), immunofluorescence staining and Western blot analysis were done to detect the PKC-α mRNA and protein expression in pulmonary arteries. The results showed that in cigarette smoke-exposed rats the percentage of CMA/IAPA and α-SM-actin expres- sion were increased greatly, PASMC apoptosis was increased and proliferation was markedly in- creased; Apoptosis indices (AI) and proliferation indices (PI) were higher than in C group; AI and PI were correlated with vascular remodeling indices; The expression of PKC-ct mRNA and protein in pulmonary arteries was significantly higher than in C group. In rats treated with puerarin, the per- eentage of CMA/IAPA and cell proliferation was reduced, whereas PASMC apoptosis was increased; The expression levels of PKC-α mRNA and protein were lower than in smoke exposure rats. There was no difference among all these data between S groups and PC groups. These findings suggested that cigarette smoke-induced pulmonary vascular remodeling was most likely an effect of the imbal- ance of PASMC proliferation and apoptosis. Puerarin appears to be able to reduce cell proliferation and vascular remodeling possibly through PKC signaling transduction pathway.     

Effect of expression of TGF-beta and TNF-alpha with Qidan granule treatment in rats by bleomycin-induced pulmonary fibrosis

Objective: To evaluate the therapeutic effects and mechanisms of Qidan granule in blemycinA5-induced pulmonary interstitial fibrosis (PIF)in rats. Methods: PIF models were established by blemycinA5-induced in rats. They were treated by Qidan granule and Hydrocortisone respectively. The pathological changes and collagen protein disposition were observed, and the expression of TGF-β, TNF-α proteins were measured by immunohistochemical technique . Results: The pulmonary alveolitis and fibrosis were alleviated remarkably in Qidan granule group compared with those in the model control group and hydrocortisone group (P <0. 01). The expression of TGF-β and TNF-α protein were higher in Qidan granule group than those in normal group , and were significantly less than those in the model control group and in hydrocortisone group (P < 0. 01). Conclusion: Qidan granule would ameliorate the pulmonary alveolitis and fibrosis. TGF-β and TNF-α might play an important role in the development of alveolitis and fibro