Iran bears a remarkable variety of reptiles. One of the lizard families occurring in Iran is the Family Agamidae which is widely are distributed throughout the old world. The large-scaled rock agamid, Laudakia nupta, is one of the well-known agamid. There are few reports of cloacal microbial on reptiles hence their function in cloacae remains unknown. Laudakia nupta usually live in rural and urban areas and close vicinity to man, they are likely to play an important role in the spread of disease that may be caused by these microorganisms and their transmission to man. Therefore, the aim of this study was to identify the bacterial flora colonizing the cloacal region of Laudakia nupta using molecular studies. The cloacal fluids were directly placed on nutrient agar (NA) plates and incubated at 25 ± 2 ℃ for 48 h. The resulting bacterial colonies were transferred to fresh nutrient agar (NA) plates for molecular studies. Twelve isolates were obtained from 17 specimens of Laudakia nupta. All bacteria isolates were identified as Bacillus subtillis (5), Bacillus cereus (4), Bacillus sp. (1), Pseudomonas putida (1), and Pseudomonas sp. (1) based on partial sequences of the 16 s rRNA gene. This is the first comprehensive report of bacteria spp. associated with cloaca of Laudakia nupta using molecular studies. In this research, we found that Laudakia nupta can be a carrier of bacteria which can transfer microorganisms to hosts.
Agar ; Bacillus ; Bacillus cereus ; Bacteria ; Cloaca ; Colon ; Genes, rRNA ; Humans ; Iran ; Lizards ; Pseudomonas ; Pseudomonas putida ; Reptiles

Epimorphic regeneration in vertebrates involves the restoration of lost tissue or organs through the formation of a regeneration blastema and occurs through a complex interaction of a number of molecular signaling pathways. Of the many effectors of successful tail regeneration in the lizard Hemidactylus flaviviridis, one crucial pathway is the cyclooxygenase-2 (COX-2) mediated PGE2 signaling pathway. The current study was aimed at understanding whether COX-2 signaling plays any role in the expression of Wnt/β-Catenin signaling components during regenerative outgrowth in H. flaviviridis. Etoricoxib—selective inhibitor of the inducible isoform of COX-2—was administered to lizards orally. We tested the expression of β-Catenin during wound epidermis and blastema stages in the regenerating tail and found a reduction in its expression in response to drug treatment. Further, it was observed that the expression of canonical Wnt ligands was greatly altered due to COX-2 inhibition. Our results provide evidence of a cross-talk between the COX-2 induced PGE₂ pathway and Wnt/β-Catenin signaling in the regenerating lizard tail. An understanding of the interaction among various signaling pathways will help elucidate the mechanism underlying epimorphosis in lizards, the only amniotes capable of appendage regeneration.
beta Catenin ; Cyclooxygenase 2* ; Dinoprostone ; Epidermis ; Inflammation ; Ligands ; Lizards* ; Prostaglandin-Endoperoxide Synthases ; Regeneration ; Tail* ; Vertebrates ; Wounds and Injuries

PURPOSE: Immunotherapy is one of the treatment strategies for breast cancer, the most common cancer in women worldwide. In this approach, the patient's immune system is stimulated to attack microscopic tumors and control metastasis. Here, we used interferon γ-induced protein 10 (IP-10), which induces and strengthens antitumor immunity, as an immunotherapeutic agent. We employed Leishmania tarentolae, a nonpathogenic lizard parasite that lacks the ability to persist in mammalian macrophages, was used as a live delivery system for carrying the immunotherapeutic agent. It has been already shown that arginase activity, and consequently, polyamine production, are associated with tumor progression. METHODS: A live delivery system was constructed by stable transfection of pLEXSY plasmid containing the IP-10-enhanced green fluorescent protein (IP-10-egfp) fusion gene into L. tarentolae. Then, the presence of the IP-10-egfp gene and the accurate integration location into the parasite genome were confirmed. The therapeutic efficacy of IP-10 delivered via L. tarentolae and recombinant pcDNA-(IP-10-egfp) plasmid was compared by determining the arginase activity in a mouse 4T1 breast cancer model. RESULTS: The pcDNA-(IP-10-egfp) group showed a significant reduction in tumor weight and growth. Histological evaluation also revealed that only this group demonstrated inhibition of metastasis to the lung tissue. The arginase activity in the tissue of the pcDNA-(IP-10-egfp) mice significantly decreased in comparison with that in normal mice. No significant difference was observed in arginase activity in the sera of mice receiving other therapeutic strategies. CONCLUSION: Our data indicates that IP-10 immunotherapy is a promising strategy for breast cancer treatment, as shown in the 4T1-implanted BALB/c mouse model. However, the L. tarentolae-(IP-10-EGFP) live delivery system requires dose modifications to achieve efficacy in the applied regimen (six injections in 3 weeks). Our results indicate that the arginase assay could be a good biomarker to differentiate tumoral tissues from the normal ones.
Animals ; Arginase ; Breast Neoplasms ; Chemokine CXCL10 ; Female ; Genetic Therapy* ; Genome ; Humans ; Immune System ; Immunotherapy ; Interferons ; Leishmania ; Lizards ; Lung ; Macrophages ; Mice ; Neoplasm Metastasis ; Parasites ; Plasmids ; Transfection ; Tumor Burden

The high resistance to infections in lizard wounds suggests that these reptiles possess effective antimicrobial peptides in their tissues. The present immunocytochemical study shows the cellular localization of beta-defensin 27 in tail tissues and in the blood, a defensin previously identified in the lizard Anolis carolinensis through biomolecular methods. Beta-defensin-27 immunoreactivity is only observed in some large granules mainly contained in heterophilic granulocytes that are sparse within the dermis of the skin or in the isolated blood. This peptide is absent in other cell types of the skin, in keratinocytes and in subdermal muscle tissue of the tail in normal conditions. Pre-corneous keratinocytes of the regenerating tail epidermis are unlabeled or show a weak labeling for the peptide only in sparse cytoplasmic areas or in the extracellular spaces among corneocytes of the wound and regenerating epidermis. The study suggests that beta-defensin 27 is normally stored in granulocytes present in the blood or in connective tissues while in the epidermis keratinocytes do not show the presence of this peptide unless these cells are stimulated from injury to produce and likely release beta-defensins.
beta-Defensins ; Connective Tissue ; Cytoplasm ; Dermis* ; Epidermis* ; Extracellular Space ; Granulocytes* ; Keratinocytes ; Lizards* ; Methods ; Muscles ; Peptides ; Reptiles ; Skin* ; Tail ; Wounds and Injuries*

To identify some medicinal animals of Lacertilia, in total 59 individuals belonging to 12 species 7 genera 3 families, we used the universal barcoding primers to sequence these species, compared with other homologous sequences (564 bp) obtaining from the GenBank and finally constructed phylogenetic trees using Neighbor-joining, Maximum parsimony and Bayesian inference, respectively. As a result, the mean content of G + C (46.5%) was lower than that of A + T (53.5%). As calculated by Kimera-2-parameter model, the whole individuals mean distance for interspecies and intraspecies was 35. 5% and 1.7%, respectively. The mean distance for interspecies was 21 times as much as that for intraspecies. The mean distance for intraspecies of Gekko swinhonis, Hemidactylus frenatus and G. gecko was greater than 2%, respectively. Further analyses suggested that geographical groups of the three species might be of different subSpecies, even species. Of course, incorporating morphological characters and other unlinked genetic markers in future studies will offer further insights into the divergence. On the basis of phylogenetic trees constructed by COI, our results indicated that the taxonomy of the category (family, genus, and species) by DNA barcoding is consistent with morphological characters. Therefore, DNA barcoding is a useful tool for both identification and phylogeny of medicinal animals of Lacertilia, particularly for nonprofessor identifying authentication of Chinese crude drugs of these species.
Animals ; DNA Barcoding, Taxonomic ; Electron Transport Complex IV ; genetics ; Lizards ; classification ; genetics ; Medicine, Chinese Traditional ; Molecular Sequence Data ; Phylogeny ; Reptilian Proteins ; genetics ; Sequence Analysis, DNA

Anti-tumor activity of the proteins from Gecko (GP) on cervical cancer cells, and its signaling mechanisms were assessed by viable cell counting, propidium iodide (PI) staining, and Western blot analysis. GP induced the cell death of HeLa cells in a dose-dependent manner while it did not affect the viability of normal cells. Western blot analysis showed that GP decreased the activation of Akt, and co-administration of GP and Akt inhibitors synergistically exerted anti-tumor activities on HeLa cells, suggesting the involvement of PI3-kinase/Akt pathway in GP-induced cell death of the cancer cells. Indeed, the cytotoxic effect of GP against HeLa cells was inhibited by overexpression of constituvely active form of Akt in HeLa cells. The candidates of the functional proteins in GP were analyzed by Mass-spectrum. Taken together, our results suggest that GP elicits anti-tumor activity against HeLa cells by inhibition of PI3-kinase/Akt pathway.
Blotting, Western ; Cell Count ; Cell Death ; HeLa Cells ; Humans ; Lizards ; Phosphatidylinositol 3-Kinases ; Propidium ; Proteins ; Uterine Cervical Neoplasms

OBJECTIVETo study the chemical constituents of Gekko swinhonis.

METHODThe compounds were isolated and purified by several column chromatographic methods with silica gel, ODS, Sephadex LH-20 and preparative HPLC, Their structures were identified by physicochemical properties and spectral data.

RESULTThirteen compounds were isolated and elucidated as cyclo-(Pro-Leu) (1), cyclo-(Ala-Pro) (2),cyclo-(Gly-Pro) (3),cyclo-(Ala-Val) (4),6-amino-9-beta-D-ribofuranosyl-9H-purine (5), uridine (6), thymidine (7),hypoxanthine-9-beta-D-ribofuranosid (8),L-phenylalanine (9),5alpha-cholest-3,6-dione (10),cholesterol (11), 1-O-hexadecanolenin (12),octadecanoic acid (13),respectively.

CONCLUSIONAll compounds were isolated from G. swinhonis for the first time.

Animals ; Dipeptides ; analysis ; Lizards ; Medicine, Chinese Traditional

OBJECTIVETo observe the anti-tumor activity of gecko alcohol extract in vitro and in vivo.

METHODin vitro, the inhibitory effect of gecko alcohol extract on proliferation of human esophageal carcinoma EC9706 cells was measured by MTT colorimetric assay. Morphological change of apoptotic cells was observed by Hoechst33258 fluorescence staining and apoptosis induced by gecko alcohol extract was evaluated in TUNEL assay. The expression of apoptosis protein Bax and Bcl-2 in EC9706 cells was investigated by immunohistochemistry. in vivo, the inhibitory effect of gecko alcohol extract on tumor growth was examined on S180 sarcoma model.

RESULTAfter gecko alcohol extract (6-8 g x L(-1)) treatment for 24, 48 and 72 h separately, EC9706 cell proliferation was significantly inhibited in both dose- and time- dependent manner (P < 0.01). Cell apoptosis in gecko alcohol extract-treated group (6, 7 g x L(-1)) was significantly higher than that in control group [(12.73 +/- 3.84)%, (9.80 +/- 2.32)% vs. (5.87 +/- 2.54)%, P < 0.05, P < 0.01]. Although the level of Bcl-2 did not change significantly, the expression of Bax was rembarkably up-regulated in gecko alcohol extract treated group. Gecko alcohol extract inhibited the growth of S180 sarcoma in Kun-ming mice at all doses (0.6, 1.2, 2.4 g x kg(-1)) of administration. The inhibitory rate was 44.88%, 63.94% and 69.53% respectively.

CONCLUSIONFrom tumor inhibitory test, gecko alcohol extract shows significantly inhibitory effect in vitro and in vivo. The increased Bax/Bcl-2 ratio is mechanistically linked to the gecko alcohol extract-induced tumor cell apoptosis.

Animals ; Antineoplastic Agents ; chemistry ; pharmacology ; therapeutic use ; Apoptosis ; drug effects ; Cell Line, Tumor ; Ethanol ; chemistry ; Humans ; Lizards ; metabolism ; Male ; Medicine, Chinese Traditional ; methods ; Mice ; Sarcoma ; drug therapy ; bcl-2-Associated X Protein ; metabolism

Extracellular recording is a key technology to study many types of brain functions. However the amplitudes of extracellular spikes are typically about 100 muV and should be picked up and amplified specially, then recorded and analyzed online. Most of the equipment for extracellular recording are either very expensive, or too large for recording in the brains of freely moving small animals. In this paper we developed a practical multi-electrode recording system based on virtual testing technology. The system includes microelectrode amplifier, analog input device NI4472 with 24-bit resolution, and data processing software SPKrec. The features of the hardware are low noise, high input impedance, a total gain of 2 000, and large dynamic range. The software developed in LabVIEW is capable of online analysis, data recording, spike detection, fire rate histograms, etc. The system functions have been verified in extracellular recording in the olfactory bulb of Gekko gecko. The result indicates that the system with performance comparable to that of commercial products meets the requirements of recording experiments.
Animals ; Brain ; physiology ; Lizards ; physiology ; Microelectrodes ; Olfactory Bulb ; physiology

AIMTo observed the expression of estrogen receptor (ER alpha and ER beta) in the heart of Gekko swinhonis.

METHODSThe immunohistochemical technique for the estrogen receptor was used.

RESULTSThe positive ER alpha and beta cells existed in cardiac myocytes and fibroblasts of the atria and the ventricles of Gekko swinhonis and had no sexual difference. The difference of ER alpha between the atria (11.56 +/- 1.67) and ventricles (6.68 +/- 1.88) was observed in both sexes (P < 0.01).

CONCLUSIONThe atria are probably the main target tissue of estrogen through ER alpha pathway while some functions of whole heart will be regulated by estrogen through ER beta pathway. The sexual differences aren' t related to the content of ER. It may be involved in the state of activity and function of ER under the physiological conditions.

Animals ; Estrogen Receptor alpha ; metabolism ; Estrogen Receptor beta ; metabolism ; Female ; Immunohistochemistry ; Lizards ; physiology ; Male ; Myocardium ; metabolism