OBJECTIVES: To assess the absorption of α-tocopherol acetate and 18β-glycyrrhetinic acid, which are used as active ingredients in toothpaste, into a reconstructed gingival tissue. METHODS: EpiGingival™ tissues were treated with a 25% slurry of toothpaste containing 2% α-tocopherol acetate and 0.3% 18β-glycyrrhetinic acid, for 2 minutes. The treatment was repeated up to 6 times, with 1 hour intervals. After completion of all treatments, the active ingredients in the tissue extracts and receiver solutions were measured by high performance liquid chromatography. RESULTS: Although α-tocopherol acetate was not detected, α-tocopherol was detected in the tissue extracts, indicating that α-tocopherol acetate was bioconverted to α-tocopherol after absorption. We could detect 18β-glycyrrhetinic acid both in the tissue extracts and in the receiver solutions, with a positive correlation to the number of treatments. CONCLUSIONS: We found that our toothpaste effectively delivered α-tocopherol acetate and 18β-glycyrrhetinic acid to a reconstructed gingival tissue in vitro.
Absorption* ; Chromatography, Liquid ; In Vitro Techniques* ; Periodontal Diseases ; Tissue Extracts ; Toothpastes

SUMMARY: The present study was performed to analyze the expression of LH genes in the rat ovary. Expression of LH subunit genes in the rat ovary was demonstrated by amplification of ovarian RNA by RT-PCR. The ovarian LHbeta transcripts contained at least two parts of the published cDNA structure, the pituitary exons 1, 2 and 3 and the part of testicular exon 1 in the major trancripts form in rat testis. Using RIA, significant amount of LH-like molecules were detected in crude ovarian extracts, and the competition curves with increasing amount of tissue extracts were parallel with those of standard peptide, indicating that the ovarian immunoreactive LH-like material is similar to authentic pituitary LH molecule. The administration of PMSG to immature rats resulted in a sharp decrease of the ovarian LH contents after 24 h post-injection. In conclusion, these findings demonstrate that genes for LH subunits are expressed in the rat ovary, and suggest that LH can play a central role in regulation of female reproduction with both endocrine (by pituitary LH) and auto- and/or para-crime (by ovarian LH) manner.
Animals ; DNA, Complementary ; Exons ; Female ; Humans ; Lutein* ; Luteinizing Hormone* ; Ovary* ; Rats* ; Reproduction ; RNA ; Testis ; Tissue Extracts

OBJECTIVETo identify the active material of anti-hepatic fibrosis from Amydae Carapax.

METHODSMembrane separation technology was adopted to screen active fraction in Amydae Carapax, and the active components were isolated from the active fraction using gel chromatography and high performance liquid chromatography. The purified active components in Amydae Carapax were further analyzed using 4700 series time-of-flight mass spectrometer.

RESULTSProteins and peptides of Amydae Carapax with molecular weight less than 6000 were proved to have biological activity. 8 components (Bj1-Bj8) were isolated from the active fraction. Bj4, Bj6 and Bj7 were screened as active components. Bj7 was further purified, resulting in 7 components (Bj701-Bj707). Bj704 and Bj707 showed significant biological activity. Mass spectrometry showed three molecular ion peaks with highest abundance, i.e. m/e 526, 542 and 572, i.e. m/e 526, 542 and 572, in Bj707 -A The amino acid sequences of above three peptide compounds were NDDY (Asn-Asp-Asp-Tyr), NPNPT (Asn-Pro-Asn-Pro-Thr), and HGRFG (His-Gly-Arg-Phe-Gly), respectively. And M572 was the most abandunt components.

CONCLUSIONThree active peptide compounds of anti-hepatic fibrosis of Amydae Carapax were identified.

Animals ; Cell Line ; Humans ; Liver Cirrhosis ; Medicine, Chinese Traditional ; Tissue Extracts ; isolation & purification ; pharmacology

AIM: Artemisia amygdalina Decne. (Asteraceae) is a critically endangered and endemic herb of Kashmir Himalayan sub-alpine region and Pakistan. Scientific research throughout the world has evidence to support the tremendous medicinal utility of the genus Artemisia. The natural resources of medicinal plants are being reduced day by day. This study provides the alternative way for medicinal resource utilization and conservation of A. amygdalina. METHODS: In vitro-raised plants and greenhouse acclimatized plants were obtained by culturing wild explants on Murashige and Skoog's medium. Plant extracts were obtained and subjected to different antioxidant assays: DPPH assay, riboflavin photo-oxidation assay, deoxy ribose assay, ferric thiocyanate assay, thiobarbituric acid assay, post mitochondrial supernatant assay and DNA damage on agarose gel. RESULTS: In vitro grown plants, as well as those acclimatized in the greenhouse reveals antioxidant activity against hydroxyl, superoxide, and lipid peroxyl radicals. CONCLUSION This preliminary study revealed the free radical scavenging potential of tissue culture-raised plant extracts of A. amydalina.
Acclimatization ; Artemisia ; chemistry ; growth & development ; physiology ; Free Radical Scavengers ; chemistry ; Plant Extracts ; chemistry ; Tissue Culture Techniques

OBJECTIVETo evaluate the influence of adipose tissue extract on inducing angiogenesis and adipogenesis in adipose tissue engineering chamber in vivo.

METHODS6 months' healthy New Zealand rabbits (n = 64) were picked. The inguinal fat pads were cultured, centrifuged, filtered, and the liquid was called adipose tissue extract (ATE). Two adipose tissue engineering chamber were built in the rabbit's back. A week later, 0.2 ml normal saline (control group, left) and 0. 2 ml ATE (experimental group, right) was respectively injected into the chamber. The contents were evaluated morphometrically, histologically and immunohistochemically 3 days, 1 week, 2 weeks, 3 weeks, 4 weeks and 7 weeks after injection. 8 rabbits were observed each time. The data regarding the number of the volume of fat flap and blood capillary at each time point were analyzed by paired t test.

RESULTSAfter injection, new tissue volume was significantly increased in the experimental group [(5.12 ± 0.22) ml], compared with that in control group [(4.90 ± 0.15) ml]. Early angiogenesis was also increased after ATE injection and the total number of capillaries reached peak 1 week after injection, which was (72.80 ± 9.67) in experimental group and (51.40 ± 6.09) in control group. In the mid-term of experimental period, earlier adipogenesis appeared in experimental group. In the later period, the outer capsule of the new construction was thinner in experimental group which reduced the suppression of the adipogenesis.

CONCLUSIONSATE can promote the angiogenesis and adipogenesis in the chamber, and reduce the capsule contracturing, so as to induce the large volume of adipose tissue regeneration

Adipogenesis ; drug effects ; physiology ; Adipose Tissue ; chemistry ; physiology ; Animals ; Neovascularization, Physiologic ; drug effects ; Rabbits ; Regeneration ; Tissue Engineering ; instrumentation ; Tissue Extracts ; pharmacology

Recent studies clearly demonstrated that the novel expression of LH gene in the rat testis, and suggested the local action of the LH-like molecule. The present study was performed to analyze the expression of the LH genes in the rat accessory reproductive organs. Expression of LH subunit genes in the rat uterus and epididymis was demonstrated by reverse transcription-polymerase chain reaction (RT-PCR) and specific LH radioimmunoassay (RIA). The LHbeta transcripts in these organs contained the published cDNA structure, the pituitary type exons 1-3, which encoded the entire LHbeta/ polypeptide. Presence of the transcripts for the alpha-subunit in the rat reproductive tissues were also confirmed by RT-PCR. In the LH RIA, significant levels of LH were detected in crude extracts from the rat ovary, uterus and epididymis. The competition cuties with increasing amount of tissue extracts were parallel with those of standard peptide, indicating that the immunoreactive LH-like materials in these tissues are similar to authentic pituitary LH molecule. In rat epididymis, the highest amount of immunoreactive LH was detected in corpus area. Our findings demonstrated that the genes for LH subunits are expressed in the rat accessory reproductive organs, and suggested that these extrapituitary LH may act as a local regulator with auto and/or paracrine manner.
Animals ; Complex Mixtures ; DNA, Complementary ; Epididymis* ; Exons ; Female ; Lutein* ; Luteinizing Hormone* ; Male ; Ovary ; Radioimmunoassay ; Rats* ; Testis ; Tissue Extracts ; Uterus*

OBJECTIVETo study the pharmacokinetics and biodistribution of pur derivative nanopaticles 4ac in mice.

METHODAfter intravenous administration of 15 mg x kg(-1) of pur derivative 4ac, the plasma and tissue concentration was detected by HPLC. The pharmacokinetics parameters were calculated by the method of 3p97.

RESULTThe concentration time curves were conformed to two compartment model. The Re of 4ac nanopaticles was 21.18 times higher than suspension.

CONCLUSIONOur present study demonstrates that, compared to nanopaticles and suspension, nanopaticles significantly alter the pharmacokinetics in plasma and tissues targeting. Biodistribution of pur derivative 4ac nanopaticles in heart of mice was better than 4ac suspension. It will become a new drug for cardiovascular disease therapy.

Animals ; Female ; Isoflavones ; chemistry ; pharmacokinetics ; Male ; Mice ; Nanoparticles ; chemistry ; Plant Extracts ; chemistry ; pharmacokinetics ; Pueraria ; chemistry ; Random Allocation ; Tissue Distribution

Amyloid fibril formation has been implicated in the pathogenesis of neurodegenerative diseases. Fibrillation generates numerous conformers. Presumably, the conformers may possess specific biological properties, thus providing a biochemical framework for strains of prions. However, the precise relationship between various fibril conformers and their pathogenic functions has not been determined because of limited accessibility to adequate amounts of fibrils from tissue samples. α-Synuclein is one such protein, and it has been implicated in Parkinson disease. Using a technique known as protein misfolding cyclic amplification, originally developed for amplifying prions, we established a procedure through which the amplification of α-synuclein fibrils is possible. With a trace amount of seeds, we succeeded in amplifying α-synuclein fibrils. The replication of the seeds was faithful in terms of conformation even after multiple rounds of cyclic amplification. Moreover, two transgenic mouse strains each representing a distinct synucleinopathy were used to investigate different conformers by using this technique. The amplified α-synuclein fibrils derived from the tissue extracts of these two strains led to the production of two different fibril conformers with distinct proteinase K digestion profiles. Together, our results demonstrated that a trace amount of α-synuclein fibrils in tissue extracts could be amplified with their conformations conserved. This procedure should be useful in amplifying α-synuclein fibrils from the brains and body fluids of patients afflicted with synucleinopathies and may serve as a potential diagnostic tool for Parkinson disease and other synucleinopathies.
Amyloid ; Animals ; Body Fluids ; Brain ; Digestion ; Endopeptidase K ; Humans ; Mice ; Mice, Transgenic ; Neurodegenerative Diseases ; Parkinson Disease ; Prions ; Tissue Extracts

OBJECTIVETo study the changes of thrombomodulin(TM) in both plasma and tissue extracts of cancer patients for evaluating its clinical significance.

METHODSPlasma TM levels were measured by enzyme-linked immunosorbent assay(ELISA) in plasma of 188 cancer patients and in 24 cancer tissue extracts including their adjacent normal tissue.

RESULTSThe plasma TM levels both in cancer patients and in metastasis patients were significantly higher than that in controls [(33.47+/-14.25) microg/L/ (41.68 +/-16.96) micro/L, compared with (20.40+/-7.22) microg/L, P<0.01]. The plasma TM levels in cancer patients after operation decreased obviously [(18.45+/-9.96) microg/L, compared with (28.29+/-11.74) microg/L,P<0.01]. Whereas, the plasma TM levels in patients with recurrence and metastasis after operation increased obviously [(34.50+/-12.57 micro/L]. The plasma TM levels in metastasis of lung cancers, gastric cancers and pancreatic cancers were significantly higher than that in non-metastasis (P<0.05 approximate, equals 0.01) respectively, but no significant differences were found between controls and non-metastasis cancers including gastric cancers, pancreatic cancers, nasopharyngeal cancers, large intestine cancers and laryngeal cancers (P>0.05). The TM levels in cancer tissue extracts were significantly lower than that in their adjacent normal tissue extracts [(647.71+/-317.51)) microg/L,compared with (1455.63+/-772.22) microg/L,P<0.01]. On the contrary, the plasma TM levels in these cancers were higher than that in controls.

CONCLUSIONThe rise of plasma TM levels in cancer patients is associated with metastasis and diffusion of cancers. The TM levels can be used as an sensitive index for judging progression and metastasis of cancers.

Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Neoplasm Metastasis ; Neoplasms ; blood ; chemistry ; Thrombomodulin ; analysis ; blood ; Tissue Extracts ; chemistry

Animals ; Ants ; Fatigue ; prevention & control ; Liver ; enzymology ; metabolism ; Mice ; Physical Conditioning, Animal ; Swimming ; Tissue Extracts ; administration & dosage ; pharmacology