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PURPOSE To assess the potential role of amphiregulin as messenger molecule in ocular axial elongation. METHODS The experimental study included guinea pigs (total n = 78) (age: 3-4 weeks) which underwent bilateral lens-induced myopization and received 15 days later three intraocular injections in weekly intervals of amphiregulin antibody (doses:5 μg, 10 μg, 20 μg) into their right eyes, and three phosphate-buffered saline injections into their left eyes; and guinea pigs without lens-induced myopization and which received three unilateral intraocular injections of amphiregulin antibody (dose: 20 μg) or amphiregulin (doses: 1 ng; 10 ng; 20 ng) into their right eyes, and three phosphate-buffered saline injections into their left eyes. Seven days later, the animals were sacrificed. Intravitally, we performed biometry, and histology and immunohistochemistry post-mortem. RESULTS In animals with bilateral lens-induced myopization, the right eyes receiving amphiregulin antibody showed reduced axial elongation in a dose-dependent manner (dose: 5 μg: side difference: 0.14 ± 0.05 mm;10 μg: 0.22 ± 0.06 mm; 20 μg: 0.32 ± 0.06 mm; p

The prevalence of antibiotic resistance and lack of alternative drugs have posed an increasing threat to public health. Here, we prepared β-Ga2O3:Cr3+ nanoparticles modified with ICAM1-antibody-conjugated TPGS (I-TPGS/Ga2O3) as a novel antibiotic carrier for the treatment of drug-resistant infections. Methods: I-TPGS/Ga2O3 were firstly characterized by measuring particle size, morphology, crystal structure, drug loading capacity, and in vitro drug release behaviors. The in vitro antibacterial activities of I-TPGS/Ga2O3/TIG were evaluated using standard and drug-resistant bacteria. The internalization of I-TPGS/Ga2O3 was observed by fluorescence confocal imaging, and the expression levels of the efflux pump genes of TRKP were analyzed by real-time RT-PCR. In vitro cellular uptake and in vivo biodistribution study were performed to investigate the targeting specificity of I-TPGS/Ga2O3 using HUEVC and acute pneumonia mice, respectively. The in vivo anti-infective efficacy and biosafety of I-TPGS/Ga2O3/TIG were finally evaluated using acute pneumonia mice. Results: It was found that TPGS could down-regulate the over-expression of the efflux pump genes, thus decreasing the efflux pump activity of bacteria. I-TPGS/Ga2O3 with small particle size and uniform distribution facilitated their internalization in bacteria, and the TPGS modification resulted in a significant reduction in the efflux of loaded antibiotics. These properties rendered the encapsulated tigecycline to exert a stronger antibacterial activity both in vitro and in vivo. Additionally, targeted delivery of I-TPGS/Ga2O3 mediated by ICAM1 antibodies contributed to a safe and effective therapy. Conclusion: It is of great value to apply I-TPGS/Ga2O3 as a novel and effective antibiotic delivery system for the treatment of drug-resistant infections.

Summary Mesenchymal stem/stromal cells (MSCs) reside in the perivascular niche and modulate tissue/organ homeostasis; however, little is known about whether and how their localization and function are linked. Particularly, whether specific MSC subsets couple with and regulate specialized vessel subtypes is unclear. Here, we show that Gli1+ cells, which are a subpopulation of MSCs couple with and regulate a specialized form of vasculature. The specific capillaries, i.e., CD31hiEMCNhi type H vessels, are the preferable vascular subtype which Gli1+ cells are adjacent to in bone. Gli1+ cells are further identified to be phenotypically coupled with type H endothelium during bone growth and defect healing. Importantly, Gli1+ cell ablation inhibits type H vessel formation associated with suppressed bone generation and regeneration. Mechanistically, Gli1+ cells initiate angiogenesis through Gli and HIF-1α signaling. These findings suggest a morphological and functional framework of Gli1+ cells modulating coupled type H vasculature for tissue homeostasis and regenerative repair., Highlights • Gli1+ cells regulate the neighbored specialized vessels in bone • Upon bone defect Gli1+ cells emerge and induce type H vessel formation for healing • Gli1+ cell ablation reduces type H vessels associated with suppressed osteogenesis • Gli and HIF-1α signaling mediate Gli1+ cell regulation of endothelial angiogenesis, In this article, Jin and colleagues show that skeletal Gli1+ cells represent a subpopulation of MSCs characterized by perivascular location, which spatially and phenotypically couple with a neighbored specialized vessel subtype, i.e., type H vessels. Functionally, Gli1+ cell-mediated regulation of type H vessel formation is indispensable for bone growth and regeneration.

Clinical observations have shown that the boundary of tumor ablation is often less than safe border and that the use of radiofrequency ablation (RFA) in the treatment of hepatocellular carcinoma (HCC) may probably accelerate its recurrence and metastasis. RFA can cause the formation of a transition zone between normal liver tissues and necrotic coagulation, where blood stagnation and thrombosis expose residual cancer cells to a hypoxic microenvironment. As the blocked vessels are slowly reperfused, the oxygen supply is gradually restored. Here, HCC cells underwent heat treatment and were cultured under hypoxic conditions to mimic the aforementioned situation, and morphological changes were observed in the surviving cells. Compared with their parental cells, hypoxic HCC cells showed changes that include enhanced invasive, metastatic, and chemoresistant abilities as well as mesenchymal characteristics. There was also a higher percentage of stem-like cells. However, either improving the hypoxic microenvironment or silencing hypoxia inducible factor (HIF)-1α signaling significantly reduced the invasive, metastatic, and chemoresistant potential and reversed the epithelial-mesenchymal transition to varying degrees. Together, these results indicated that a sustained hypoxic microenvironment after RFA may exert a negative impact on the prognosis of HCC patients, and minimizing exposure to a hypoxic microenvironment and targeting HIF-1α signaling might be effective strategies for patients who experience insufficient RFA therapy.

To assess anatomical changes in eyes with progressive myopia, we morphometrically examined the eyes of guinea pigs with lens-induced axial elongation. Starting at an age of 3–4 weeks, guinea pigs in the experimental group (n = 20 animals) developed unilateral lens-induced axial elongation by wearing goggles for 5 weeks compared to a control group of 20 animals without intervention (axial length:8.91 ± 0.08 mm versus 8.74 ± 0.07 mm; P P = 0.41), equator (P = 0.41), midpoint between equator and posterior pole (MBEPP) (P = 0.13) or posterior pole (P = 0.89). Retinal pigment epithelium (RPE) cell density decreased with longer axial length at the MBEPP (P = 0.04; regression coefficient beta = −0.33) and posterior pole (P = 0.01; beta = −0.40). Additionally, the thickness of the retina and sclera decreased with longer axial length at the MBEPP (P = 0.01; beta = −0.42 and P P P P

Background/Aims: An enriched environment (EE) ameliorates learning and memory impairments induced by chronic cerebral hypoperfusion, and the p38 mitogen-activated protein kinase (p38 MAPK) signaling pathway exerts both beneficial and deleterious effects on the nervous system during the progression of ischemia. Methods: The present study investigated whether p38 MAPK participates in the process by which EE exposure ameliorates the cognitive deficits induced by chronic cerebral hypoperfusion. Results: EE exposure significantly enhanced the cognitive performance of vascular dementia (VD) model rats, and p38 MAPK protein decreased in parallel with cognitive improvements. Inhibition of p38 MAPK function by its selective inhibitor SB203580 improved the cognition index of VD rats and upregulated p38 MAPK expression with p38 MAPK antisense oligodeoxynucleotides. This impaired cognition in VD rats could not be rescued by EE exposure. Conclusion: p38 MAPK participates in the process by which EE exposure ameliorates cognitive deficits induced by chronic cerebral hypoperfusion.

Hepatocellular carcinoma is one of the leading causes of malignancy-related death in China. Its therapy in clinics is a big challenge. Ginsenoside Rh2 is one of the most notable cancer-preventing components from red ginseng and it has been reported that ginsenoside Rh2 exhibited potent cytotoxicity against human hepatoma cells. Rh2 exists as two different stereoisomeric forms, (20S)-ginsenoside Rh2 and (20R)-ginsenoside Rh2. Previous reports showed that the Rh2 epimers demonstrated different pharmacological activities and only (20S)-ginsenoside Rh2 showed potent proliferation inhibition on cancer cells in vitro. However, the in vivo anti-hepatoma activity of (20R)-ginsenoside Rh2 and (20S)-ginsenoside Rh2 has not been reported yet. This work assessed and compared the anti-hepatoma activities of (20S)-ginsenoside Rh2 and (20R)-ginsenoside Rh2 using H22 a hepatoma-bearing mouse model in vivo. In addition, hematoxylin and eosin staining, the deoxynucleotidyl transferase dUTP nick-end labeling assay, and the semiquantitative reverse transcriptase polymerase chain reaction method were used to further study the apoptosis of the tumors. The results showed that both (20S)-ginsenoside Rh2 and (20R)-ginsenoside Rh2 suppressed the growth of H22 transplanted tumors in vivo, and the highest inhibition rate could be up to 42.2 and 46.8 %, respectively (p 0.05). Further, hematoxylin/eosin staining and the deoxynucleotidyl transferase dUTP nick-end labeling assay indicated that both (20R)-ginsenoside Rh2 and (20S)-ginsenoside Rh2 could induce H22 hepatoma tumor cell apoptosis, with apoptosis indexes of 3.87 %, and 3.80 %, respectively (p 0.05). Moreover, this effect was accompanied by downregulating the level of Bcl-2 mRNA. In conclusion, both (20S)-ginsenoside Rh2 and (20R)-ginsenoside Rh2 can suppress the growth of H22 hepatomas without causing severe side effects, and this effect is associated with the induction of apoptosis.

Ferroptosis, a newly discovered form of non-apoptotic cell death, is induced by an excessive degree of iron-dependent lipid peroxide. ATPR, a novel all-trans retinoic acid (ATRA) derivative, has been extensively developed to show superior anticancer effect than ATRA in acute myeloid leukemia (AML). However, whether ferroptosis exists during ATPR treatment of AML remains unclear. Herein, we found that ferroptosis occurred in an AML xenograft mouse model of ATPR treatment. In vitro, ATPR was verified to induce ferroptosis in a dose-dependent manner by proferroptotic protein marker, lipid peroxidation, and lipid ROS, which could be significantly reversed by ferrostatin-1. Using lysosomal inhibitor chloroquine and iron chelator desferrioxamine, we further revealed that ATPR-induced ferroptosis was regulated by autophagy via iron homeostasis, especially Nrf2. Furthermore, targeting ferroptosis contributes to ATPR-induced AML differentiation. In conclusion, these results indicated that ferroptosis play an important role in ATPR-induced differentiation, and suggested that ATPR would provide a potential therapeutic value for AML treatment.

Salidroside administration improves memory in different models of learning. However, its influence on models of Alzheimer's disease (AD) has not been widely studied. In the present study, the therapeutic effect of salidroside was investigated in an animal model of AD. APPswe/PS1ΔE9 mouse (n=20) were randomly divided into either the AD model group or the salidroside + AD model group (n=10 in each group), and C57BL/6J mouse (n=20) of identical age and genetic background were randomly divided into either the normal control (NC) group or the salidroside + NC group (n=10 in each group). The Morris water maze behavioral test was applied to all mice in order to investigate the effects of salidroside administration on learning and memory functions. The concentrations of malondialdehyde (MDA), glutathione (GSH) and nitrate in the hippocampus of the mice were determined, and hippocampal superoxide dismutase (SOD) activity was also determined. In addition, terminal deoxynucleotidyl‑transferase‑mediated dUTP nick end labeling was used to investigate the rate of neuronal apoptosis in the hippocampus. Furthermore, the concentrations of interleukin‑6 (IL‑6) and tumor necrosis factor‑α (TNF‑α) were tested for in the brain tissues of AD mice. Learning and memory functions in AD mice were revealed to improve following administration of salidroside. Furthermore, salidroside administration was revealed to decrease the concentrations of MDA and nitrate in the hippocampus, decrease the apoptotic rate of hippocampal neurons, and increase the activity of SOD and the concentration of GSH in hippocampal tissue. In addition, it was demonstrated that salidroside administration suppressed the expression levels of IL‑6 and TNF‑α. In conclusion, this study revealed that the administration of salidroside could attenuate the effects of AD‑associated memory and learning impairment in mice. Furthermore, it was demonstrated that the effects of salidroside administration on AD mice were, at least partially, via inhibition of brain oxidative/nitrosative damage, suppression of both IL‑6 and TNF‑α expression levels, and suppression of the hippocampal neuronal apoptotic rate.

To study the effect of enamel matrix derivative(EMD) on root repair and tooth movement relapse after orthodontic tooth movement in rats.Twenty 10-weeks old male rats were divided into experimental and control group randomly (n=10). 100 g force was exerted to the left maxillary first molars for 14 days and then removed. EMD was injected in rats in the experimental group since appliance was removed. The control group received no injection. Micro-CT was taken to analyze the changes of root resorption volume and distance of tooth movement. The differences were analyzed by SPSS19.0 software package.Fourteen days after device removal, the repair volume of root resorption lacunae in the experimental group and the control group were (0.0295±0.0052)×10EMD can enhance the effect of root repair and decrease early relapse after orthodontic tooth movement in a certain extent.

Renal ischemia-reperfusion injury (IRI) is regarded as a leading cause of acute kidney failure and renal dysfunction. Previous studies show that kappa opioid receptor (KOR) agonists can attenuate IRI in cardiomycytes and neuronal cells. In this study we explored the effects of a KOR agonist on renal IRI and the underlying mechanisms in vivo and in vitro. An IRI model was established in SD rats, which were intravenously pretreated with a KOR agonist U50448H (1 mg/kg), a KOR antagonist Nor-BNI (2 mg/kg) followed by U50448H (1 mg/kg), or the PI3K inhibitor wortmannin (1.4 mg/kg) followed by U50448H (1 mg/kg). U50448H pretreatment significantly decreased the serum levels of creatinine (Cr) and BUN, the renal tubular injury scores and the apoptotic index (AI) in IRI model rats. Furthermore, U50448H significantly increased SOD activity and NO levels, and reduced the MDA levels in the kidney tissues of IRI model rats. Moreover, U50448H significantly increased the phosphorylation of Akt, eNOS and PI3K in the kidney tissues of IRI model rats. All the beneficial effects of U50448H were blocked by Nor-BNI or wortmannin pre-administered. Similar results were observed in vitro in renal tubular epithelial NRK-52E cells subjected to a hypoxia-reoxygenation (HR) procedure. Our results demonstrate that the KOR agonist U50448H protects against renal IRI via activating the PI3K/Akt signaling pathway.

Exposure to fenvalerate has been shown to be associated with decreased steroid hormone production by mouse Leydig tumor cells (MLTC-1) in our previous study and the interference with cAMP-PKA pathway cannot explain this inhibitory effect completely. In this study, the same cell line was used to investigate the potential involvement of insulin-like growth factor I (IGF-I) signaling pathway in the downregulation of steroidogenesis by fenvalerate. Results showed that fenvalerate treatment decreased IGF-I secretion significantly which was consistent with the reduced expression of IGF-I mRNA. Then inhibitors of the two downstream pathways of IGF-I were added to the medium. The addition of LY294002 (inhibitor of phosphatidylinositol (PI)-3-kinase) did not alter the declining trend of progesterone production with increasing dosages of fenvalerate treatment while the addition of UO126 (inhibitor of extracellular signal-regulated kinases 1/2 (ERK1/2)) markedly attenuated this trend, which strongly indicated the possible involvement of pathway ERK1/2. In addition, phosphorylation of ERK1/2 was also suppressed by fenvalerate. The results suggest that the mechanism by which fenvalerate decreased steroid hormone production might involve the impairment of IGF-I signal pathway by attenuating the IGF-I production and ERK1/2 phosphorylation.

To detect the expression of growth factors relating to bone reconstructions in microscrew-bone interface of implanted titanium microscrews near the extraction wounds, and to determine the influence of healing process on the growth factors.Eight male Beagle dogs (age 18 months) were divided into experimental and control groups. Titanium microscrews were implanted near an extraction wound in the dogs in the experimental group, while the controls had implantation without extraction wounds. The dogs were sacrificed at 1, 3, 8, 12 weeks, respectively. Immunohistochemistry and in situ hybridization sections were performed to detect the expression of TGF-beta, TNF-alpha, osteocalcin (OC) protein and Cbfal mRNA.The experimental group had higher levels of expression of TGF-beta, TNF-alpha protein and Cbfa1 mRNA than the controls, with significant differences appearing at week one for TNF-alpha (P0.05), week three for TGF-beta and TNF-alpha (P0.05), and week eight for Cbfa1 mRNA (P0.05). The expression of OC increased over time and reached peak at week eight (P0.05).Microscrews implanted near extraction wounds can maintain stability. Severe inflammation occurs in the early stage of healing, but does not sustain. Bone remodeling remains active over the healing process. But prolonged healing phase without force loading could weaken the remodeling.

Mitochondrial genome plays a central role in aging, cancer, apoptosis and metabolism. Squamous cell carcinoma is one of the major types of non–small cell lung cancer. We sequenced a complete mitochondrial genome sequence of a rat squamous cell carcinomas cell tissue for the first time. The total length of the mitochondrial genome was 16,319 bp, with 13 protein-coding genes, two ribosomal RNA genes and 22 transfer RNA genes. This genome describing information will supply the potential use of mtDNA mutations as markers in cancer.

The diagnosis of Schistosoma japonicum (Sj) infection in low prevalence areas of the People's Republic of China is challenging due to the sensitivity of the detection methods, leading to an underestimation of the disease burden. We compared the sensitivities of the miracidium hatching test (MHT) with the modified Kato-Katz method (KK) and the combination of the two methods (KK-MHT) to detect Sj infection in low prevalence areas of China. The stool samples of 3,853 residents from 8 villages with a light to moderate prevalence (0-23%) of Sj infection were examined by KK, MHT and KK-MHT. The findings were inconsistent. The KK-MHT conbination gave more positives than either the KK or MHT alone. Using the KK-MHT, we determined the missed rates with the KK (mR(K)) and MHT (mR(H)) to be 30.1% and 10.2%, respectively. At light prevalence sites (infection rate10%) the mR(K) was 60.6%, significantly higher than the mR(K) of 22.3% found at moderate prevalence sites (10-23%). However, the mR(H) at the light and moderate prevalence sites were 11.54% and 9.90%, respectively (p0.05). The combination KK-MHT had the best sensitivity in low Sj prevalence areas in China and the KK method alone was the least sensitive. Using KK alone as a screening method will result in an underestimation of Sj infection disease burden.

Schistosomiasis is a tropical disease, which could do serious damage to the people's health, and it hinders the development of the social economy but may be neglected. After a positive control, some countries and regions have blocked the spread of schistosomiasis. However, in the past few years, with the development of social economy, due to the global movement of people, schistosomiasis not only poses a threat to control areas, but also may cause new endemic areas. This article reviews the parasitological characteristics, clinical manifestations, epidemiological situation, and control approaches of three major kinds of human schistosomiasis, schistosomiasis japonica, schistosomiasis haematobia, and schistosomiasis mansoni.

The global impact of neglected tropical diseases has been much more recognized recently. The elimination and eradication of these diseases is significant for achieving the millennium development goals, for societies and for global healthcare. WHO already drew the roadmap to accelerate the work on eliminating and eradicating the selected diseases for 2015 and 2020 respectively. The present review demonstrates the current situation and progress of controlling these neglected tropical diseases and the challenges faced, and emphasizes the action of China.

To explore the impact of the impoundment of Three Gorges reservoir on Oncomelania snail habitats of Poyang Lake.Four counties with schistosomiasis endemic around the Poyang Lake, naming as Xingzi, Duchang, Jinxian and Xinjian, were selected as study areas. The average daily water level of four major hydrological stations near the above 4 counties from 1997 to 2008 were collected, as well as the elevation of 215 marshlands with snails in those 4 counties, the data of schistosomiasis endemic situation and snail survey of 4 counties in the year before the impoundment of Three Gorges reservoir (2002) and the 6th year after the impoundment of Three Gorges reservoir (2008). The fluctuations of water levels of Poyang Lake, flooding day of marshland with snails as well as the changes of the indicators in snail survey and prevention and control measures before and after the impoundment of Three Gorges reservoir were compared.Before the impoundment of Three Gorges reservoir, the fluctuations of average water level of the four hydrological stations around Poyang Lake was 6.91-12.93 m; which reduced to 5.72 - 10.75 m after the impoundment. The D-value of the fluctuations of average water level was (1.06 ± 0.47) m. The date of the annual maximum water level delayed by an average of 12 d; while the date of annual minimum water level arrived an average of 12 d earlier than the impoundment of Three Gorges reservoir. The average flooding days of marshland with snails were separately (100.04 ± 42.06) d before the impoundment and (64.13 ± 22.30) d after the impoundment. The difference was statistically significant (t = 3.70, P0.05). The density of snails in the 4 counties around Poyang Lake was 0.8437/0.1 m(2) after the impoundment; declining by 31.04% from the snail density (1.2234/0.1 m(2)) before the impoundment. The density of infectious snails was 0.0014/0.1 m(2) after the impoundment; dropping by 67.34% from it (0.0042/0.1 m(2)) before the impoundment; the infection rate of snails was separately 0.34% after the impoundment and 0.16% before the impoundment. The difference was statistically significant (χ(2) = 53.25, P0.05). The coverage rate of chemotherapy increased from 4.45% (35 558/798 200) in 2002 to 9.80% (79 457/810 400) in 2008; the difference was statistically significant (χ(2) = 17 336.63, P0.05). The coverage rate of snail killing increased from 1.35% (389.04/28 824.66) in 2002 to 7.49% (2260.90/30 166.22) in 2008; the difference was also statistically significant (χ(2) = 28 824.66, P0.05).The impact of the impoundment of Three Gorges reservoir shortened the flooding days of marshland with snails and helped the prevention and control of schistosomiasis in Poyang Lake regions as the environment went against the breeding of the snails.

To study the effects of centipede extracts on H22 tumor-bearing mouse, sarcoma S180 mouse and normal mouse.Normal and tumor-bearing mouse were orally administrated by centipede extracts. Rate of restraining tumor, index of thymus and spleen were calculated after 12 days treatment. Acute toxicity testing tried to figure out In LD50 of centipede extracts.The restraining tumor rates of centipede ethanol extracts at low and medium doses were 22.2% and 17. 88%. There was no tumor restraining effect by the high dose treatment. The tumor growth of the H22 model mouse was not restrained by the centipede water extracts. There were no significant differences among the treatments in their spleen weight and spleen index. In LD50 test, the administrating dosages of centipede extracts given to the mouse were 48 times those given to patients on clinic. The result showed no mouse dead in centipede group.Centipede water extracts had no anti-tumor effect on tumor-bearing mouse. There is certain toxicity in ethanol extracts of centipede, suggesting that centipede alone for treatment of cancer needs further study.

To investigate the effects of Shengjingbao on spermatogenesis in the mouse model of oligospermia and its action mechanisms.Sixty male mice were randomly divided into 4 groups, Shengjingbao (Group 1), Vitamin E (Group 2), blank model control (Group 3) and normal blank control (Group 4). The first three groups were treated by celiac injection of cyclophosphamide for 5 successive days to make models, followed by intragastric administration of Shengjingbao and Vitamin E to Group I and 2, respectively, for 36 days. And then all the mice were sacrificed. The serum testosterone (T) level was determined by radioimmunology, and a suspension was made from the testis and epididymis of one side for sperm analysis, while the testis of the other side was sliced up and stained by HE method and TUNEL technique to detect the count of Leydig cells, the layers of spermatogenic epithelia and the apoptosis of spermatogenic cells.Compared with Group 3, the serum T level, the layers of spermatogenic epithelia and the count of Leydig cells were obviously improved and even exceeded those in Group 2. The positive expression rate of spermatogenic cell apoptosis in Group 1 was evidently lower than Group 2 and 3. The above differences were statistically significant (P0.01 or P0.05), but no significant difference was noted between Group 1 and 4.Shengjingbao can significantly increase the count of Leydig cells, elevate the T level that influences the layers of spermatogenic epithelia, and thus enhance spermatogenesis. The action mechanisms of Shengjingbao may lie in its capacity of inhibiting the apoptosis of spermatogenic cells, increasing the layers of spermatogenic epithelia and facilitating spermatogenesis. T may play a role in inhibiting the apoptosis of spermatogenic cells.

Fenvalerate is a widely used synthetic pyrethroid insecticide and is known to impede the male reproductive function. However, the mechanisms remain to be elucidated. In this study, mouse Leydig tumor cells (MLTC-1) were used to investigate the effects of fenvalerate on progesterone production. Fenvalerate treatment inhibited progesterone secretion induced by human chorionic gonadotropin (hCG), cholera toxin (CT) or forskolin and decreased cAMP levels induced by hCG, but not by CT or forskolin, which suggested a repaired site on the upstream components of G protein or G protein per se by fenvalerate in the cAMP-mediated signal pathway. Furthermore, the addition of cAMP analog, 8-Br-cAMP, could not reverse fenvalerate-suppressed progesterone synthesis, indicating that fenvalerate interfered with the downstream molecules of cAMP. In addition, fenvalerate decreased steroidogenic acute regulatory protein (StAR) mRNA and protein levels, and also profoundly inhibited the activity of P450 side chain cleavage enzyme (P450scc) which was consistent with the decreased expression of P450scc mRNA and protein in MLTC-1 cells. These results suggested that fenvalerate might inhibit progesterone production by attenuating cAMP generation and inhibiting StAR expression and P450scc activity.

Alkylphenols (APs) are widely used as important industrial materials and have attracted lots of attention because of their potential estrogenic activities. In this study, we developed human estrogen receptor alpha (hERalpha) and rat estrogen receptor alpha (rERalpha) mediated reporter gene assays and compared the estrogenic activity of APs and related chemicals based on the two ERalpha. Human breast cancer cell line MCF-7 was co-transfected with Gal4-fused hERalpha and corresponding reporter plasmid; African green monkey kidney cell line CV-1 was co-transfected with rERalpha and reporter gene. Both assays showed acceptable response to natural estrogen 17beta-estradiol (E2) with EC50 of 0.16 nM and 4.7 nM. Then the estrogenic activity of 4-APs, 4-phenylphenol and bisphenol-A were evaluated and compared with the effects of E2. The data suggested that test APs and related chemicals possessed weakly estrogenic activity and the activity of test APs increased with the increase of substituent size. This structure-activity relationship helped to infer the activity of chemicals with similar feature. Furthermore, test APs showed similar effect on the function of hERalpha and rERalpha. This consistency helped to extrapolate in vivo rodent data to human being when performing risk assessment of endocrine disruptors.

Many pesticides possess hormonal activity and have thus been classified as endocrine disruptors. Pyrethroids are commonly used pesticides worldwide, but little has been done to characterize their antiandrogenic activity potential. We tested three frequently encountered pyrethroids (fenvalerate, cypermethrin, permethrin) and their metabolite 3-phenoxybenzoic acid (3-PBA) for antiandrogenic and androgenic activity using a human androgen receptor (AR) mediated luciferase reporter gene assay in CV-1 African green monkey kidney cell. The assay displayed appropriate response to the known AR agonist 5alpha-dihydrotestosterone and AR antagonist nilutamide and flutamide. At 0.1mM, all the three tested pyrethroids significantly suppressed the luciferase expression. Further, their metabolite 3-PBA also showed antagonist activity. None of the test chemicals showed androgenic activity. Through the antiandrogenic pathways, exposure to certain pyrethroids may contribute to the damage of reproductive system. In conclusion, pyrethroid pesticides can act as antiandrogen in vitro, and metabolizing to 3-PBA cannot eliminate the antagonist activity. This result provides useful information for risk assessment of pyrethroid pesticides.

Angiogenesis is the essential process for tumor growth and metastasis. Angiostatin, a potent endogenous inhibitor of angiogenesis, could selectively inhibit proliferation of vascular endothelial cells. This study was to construct recombinant angiostatin-baculovirus, and explore the expression and biological activity of recombinant angiostatin in insect cells.The angiostatin baculovirus transfer vector pBlueBacHis2B was co-transfected with virus DNA into insect Sf9 cells to construct recombinant baculovirus. The recombinant virus was screened by plaque assay, and confirmed and amplified by polymerase chain reaction (PCR) to produce large scale, high-titer virus stock. The expression of the recombinant protein at different time points was detected by SDS-PAGE and Western blot. The recombinant protein was purified by ProBond purification system. Inhibitory effect of recombinant angiostatin on human umbilical vein endothelial cells (HUVECs) was examined by MTT assay. Its anti-angiogenesis effect was confirmed by in vivo chorioallantoic membrane (CAM) test.Recombinant angiostatin baculovirus with a high virus titer (2 x 10(8) pfu/ml) was constructed successfully. Recombinant angiostatin (53 ku) was effectively expressed in Sf9 cells, and its pure degree was about 90% of insect cellular total soluble proteins. The recombinant angiostatin obviously inhibited proliferation of endothelial cells with the 50% inhibitory concentration (IC(50)) of 2.3 microg/ml, and remarkably inhibited the growth of vessels in CAM.The baculovirus expression system could be used to construct high-titer recombinant angiostatin-virus stock, and highly express the recombinant angiostatin in insect cells. In vitro and in vivo experiments confirmed that angiostatin could inhibit proliferation of vascular endothelial cells.