In order to investigate the characteristics of transdermal delivery of ferulic acid under the treated of microneedle arrays and the influence on permeability of rat skin capillaries, improved Franz-cells were used in the transdermal delivery experiment with the rat skin of abdominal wall and the length of microneedle arrays, different insertion forces, retention time were studied in the influence of characteristics of transdermal delivery of FA. The amount of FA was determined by HPLC system. Intravenous injection Evans blue and FA was added after microneedle arrays treated. Established inflammation model was built by daubing dimethylbenzene. The amount of Evans blue in the rat skin was read at 590 nm wavelength with a Multiskan Go microplate reader. Compared with passive diffusion group the skin pretreated with microneedle arrays had a remarkable enhancement of FA transport (P <0.01). The accumulation of FA increased with the enhancement of insertion force as to as the increase of retention time. Microneedle arrays with different length had a remarkable enhancement of FA transport, but was not related to the increase of the length. The research of FA on the reduce of permeability of rat skin capillaries indicated that the skin pretreated with microneedle arrays could reduce the content of Evans blue in the skins of rat significantly compared with the untreated group. The permeation rate of ferulic acid transdermal delivery had remarkable increase under the treated of microneedle arrays and the length of microneedle arrays ,the retention time so as to the insertion force were important to the transdermal delivery of ferulic acid.
Administration, Cutaneous ; Animals ; Coumaric Acids ; administration & dosage ; pharmacokinetics ; Male ; Needles ; Rats ; Rats, Sprague-Dawley ; Skin Absorption

The stability of melittin in different solvents (water, deoxygenated water, physiological saline, PBS, 50% ethanol, ethanol, glycerol)was studied and the results showed that the stability of melittin is not influenced by light, temperature and pH in 50% ethanol, which melittin can be completed dissolved when compared with ethanol and glycerol, in such, 50% ethanol was chosen as solvent storage when measured content of melittin. Then the effect of different concentrations of PBS, the pH of PBS and rat skin ho- mogenates were tested, and the results showed that melittin was degraded rapidly at low concentration solution and low ionic strength. Increasing pH of PBS and rat skin homogenate can accelerate the degradation of melittin. These researches provide an experimental ba- sis for further study of melittin.
Animals ; Drug Stability ; Ethanol ; chemistry ; Hydrogen-Ion Concentration ; Melitten ; chemistry ; Rats ; Skin ; drug effects ; Solvents ; chemistry ; Temperature

Objective To discuss the value of diffusion-weighted MRI (DWI) and 18F-FDG-PET/CT in assessing the early therapeutic effect of radiofrequency ablation (RFA) for VX2 sarcomas in experimental rabbits. Methods VX2 sarcoma was inoculated at bilateral hind limbs in 14 New Zealand white rabbits to establish the animal models. The implanted VX2 tumor on one hind leg was treated with ultrasound-guided percutaneous RFA (study group), while no RFA was given to the VX2 tumor on the contralateral hind leg (control group). DWI-MRI was performed at 2 days after RFA, and 18F-FDG-PET/CT examination was employed at 3 days after RFA. The mean apparent diffusion coefficient (ADC) values and standard uptake value (SUV) of the untreated tumor and the ablated tumor were separately calculated. Taking the pathologic result as the gold standard, the consistency of DWI-MRI, PET/CT as well as the combination of DWI-MRI and PET/CT with the clinical diagnosis was separately evaluated by Kappa test. Results Before RFA, DWI-MRI demonstrated that the VX2 tumor was characterized by hypo-intensity signal on T1 and hyper-intensity signal on T2 with ring-shaped enhancement on T1-weighted image; PET/CT showed that the tumor had nodular or ring-shaped 18F-FDG accumulation. After RFA, DWI-MRI revealed that the VX2 tumor was manifested as hyper-intensity signal on T1 and slight higher density on T1 with slight enhancement on contrast-enhanced T1-weighted image; PET/CT showed lowered accumulation of 18F-FDG. The mean ADC value of the ablated tumor was (1.52 ± 0.24) × 10-3 mm2/s, which was obviously higher than that of the un-ablated tumor, that was (1.09 ± 0.12) × 10-3 mm2/s, the difference was statistically significant(P<0.05). The mean SUV value of the ablated tumor was (0.6 ± 0.3), which was significantly lower than that of the ablated tumor (9.6 ± 3.2, P<0.05). No significant difference in sensitivity, specificity and accuracy existed between DWI-MRI and pathology as well as between PET/CT and pathology, the Kappa value being 0.357 and 0.428 respectively (P>0.05). The Kappa value of the consistency between combination of DWI-MRI with PET/CT and pathology was 0.786, which was significantly different from the result by simple DWI-MRI or simple PET/CT evaluation (P< 0.05). Conclusion Both ADC value of DWI-MRI and SUV value of PET/CT are useful indexes for evaluating the early therapeutic effect of RFA. Both DWI-MRI and PET/CT have their respective advantages, nevertheless, combination use of both can effectively improve the evaluation of curative effect for VX2 tumor after RFA in experimental rabbits.

In order to test the equilibrium solubility of puerarin in different solvents and solubilizer,cilia toxicity and irritation of these excipient, the balance method, toad in the ciliary body toxicity and rat nasal mucosa irritation were used respectively. Results showed that puerarin solubility was 56.44 g x L(-1) in combined solvent of 30% PEG200 and 10% Kolliphor HS 15. With normal saline solution as negative control and sodium deoxycholate as positive control, the effects of 30% PEG200, 30% PEG 400, 10% Kolliphor HS 15 and combination of 30% of PEG200 and 10% Kolliphor HS 15 on toad palate cilium were observed and cilia movement duration was recorded. The results indicated that there was no significant difference in cilia movement duration among 30% PEG200, 10% Kolliphor HS 15 and normal saline group. The rats long-term nasal mucous membrane irritation of 30% PEG 400, 10% Kolliphor HS 15, which had no cilia toxicity, was studied, with normal saline solution as negative control. There were no significant difference revealed on rat nasal mucosa epithelial thickness among 30% PEG 400, 10% Kolliphor HS 15 and normal saline. Above researches showed 30% PEG 400, 10% Kolliphor HS 15 was ideal for solubility of puerarin nasal drops and showed a lower cilia toxicity and irritation, and can be used as the solvent and solubilizer of puerarin nasal drops.
Administration, Intranasal ; methods ; Animals ; Anura ; Cilia ; chemistry ; Female ; Isoflavones ; chemistry ; Male ; Nasal Mucosa ; Polyethylene Glycols ; chemistry ; Rats ; Rats, Sprague-Dawley ; Solubility ; Solvents ; chemistry

BACKGROUNDHuman amniotic epithelial cells (HAECs) are able to secrete biologically active neurotrophins such as brain-derived neurotrophic factor and neurotrophin-3, both of which exhibit trophic activities on dopamine neurons. Previous study showed that when human amniotic epithelial cells were transplanted into the striatum of 6-hydroxydopamine (6-OHDA)-induced Parkinson disease rats, the cells could survive and exert functional effects. The purpose of this study was to investigate the survival and the differentiation of human amniotic epithelial cells after being transplanted into the lateral ventricle of Parkinson's disease (PD) rats, and to investigate the effects of grafts on healing PD in models.

METHODSThe Parkinson's model was made with stereotactic microinjection of 6-hydroxydopamine (6-OHDA) into the striatum of a rat. The PD models were divided into two groups: the HAECs group and the normal saline (NS) group. Some untreated rats were taken as the control. The rotational asymmetry induced by apomorphine of the HAECs group and the NS group were measured post cell transplantation. The expression of nestin and vimentin in grafts were determined by immunohistology. Ten weeks after transplantation the density of tyrosine hydroxylase positive cells in the substantia nigra of the HAECs group, NS group and the untreated group was determined. The differentiation of grafts was determined by TH immunohistology. High performance liquid chromatography (HPLC) was used to determine monoamine neurotransmitter levels in the striatum.

RESULTSThe rotational asymmetry induced by apomorphine of the HAECs group was ameliorated significantly compared to the NS group two weeks after transplantation (P < 0.01). The grafts expressed nestin and vimentin five weeks after transplantation. TH immunohistochemistry indicated that the TH positive cells in the substantia nigra of the HAECs group increased significantly compared to the NS group (P < 0.01). Tyrosine hydroxylase (TH) positive cells in the substantia nigra of the HAEC group and the NS group were decreased compared to the untreated group (P < 0.01). Dopamine and DOPAC levels in the striatum of the HAECs group increased significantly compared to the NS group (P < 0.05). Homovanillic acid (HVA) levels in the striatum of the HAECs group increased significantly compared to the NS group (P < 0.01). In addition dopamine, DOPAC, and HVA levels in the striatum and dopamine levels in the cerebrospinal fluid of the HAECs group and the NS group were decreased compared to the untreated group (P < 0.05).

CONCLUSIONSHuman amniotic epithelial cells could be used to ameliorate the rotational asymmetry induced by apomorphine of the PD models. This could have been due to the increased content of dopamine and its metabolic products, DOPAC and HVA, in the striatum in the PD models.

Amnion ; cytology ; Animals ; Apomorphine ; pharmacology ; Chromatography, High Pressure Liquid ; Epithelial Cells ; cytology ; drug effects ; transplantation ; Female ; Homovanillic Acid ; metabolism ; Humans ; Immunohistochemistry ; Oxidopamine ; toxicity ; Parkinsonian Disorders ; chemically induced ; metabolism ; therapy ; Rats ; Rats, Sprague-Dawley

Aged ; Aged, 80 and over ; Alzheimer Disease ; blood ; immunology ; B-Lymphocytes ; immunology ; B7-1 Antigen ; blood ; CD28 Antigens ; blood ; Female ; Flow Cytometry ; HLA-DR Antigens ; blood ; Humans ; Killer Cells, Natural ; immunology ; Lymphocyte Subsets ; immunology ; Male ; T-Lymphocyte Subsets ; immunology

Objective To study the expressions of nucleostemin gene mRNA and Ki-67antigen in pituitary adenomas and investigate the role of nucleostemin gene in the tumorigenesis and development of pituitary adenomas. Methods Seventy-one samples of pituitary adenomas were collected. Semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) was used to detect the expression of nucleostemin gene mRNA in samples. Immunohistochemistry technique was applied to examine Ki-67 antigen expression in the paraffin sections of samples. At the same time,coherent clinical data were collected. Results Nucleostemin gene mRNA was detectable in all samples of pituitary adenomas. Between invasive and noninvasive pituitary adenomas, the difference of nucleostemin expression was extremely significant (P＜0.01), and Ki-67 labeling index (LI) was also significantly different (P＜0.05). The difference of Ki-67LI between recurrent and non-recurrent groups was significant (P＜0.05). There was a positive correlation between nucleostemin gene and Ki-67LIlevels (r=0.237, P＜0.05). Conclusions Nucleostemin gene plays an important roles in the invasion of human pituitary adenoma. Expression of nucleostemin gene is positively related to Ki-67 antigen expression, and both may be the valid clinical detection markers for assessing proliferation, invasion and recurrence of pituitary adenomas.

OBJECTIVETo investigate the expression of CD147, matrix metalloproteinase (MMP)-2, MMP-9, Transforming growth factor (TGFbeta1) and TGFbetaRI proteins and their relationships to breast cancer.

METHODSThe expression of CD147, MMP-2, MMP-9, TGFbeta1 and TGFbetaRI proteins was examined on tissue chips containing 160 cases of breast carcinomas by S-P immunohistochemical method.

RESULTSThe positive rates of CD147, MMP-2, MMP-9, TGFbeta1 and TGFbetaRI proteins were 87.5% (140/160), 96.9% (155/160), 95.0% (152/160), 73.7% (118/160) and 60.6% (97/160), respectively. The expression of CD147 was positively correlated with axillary lymph node metastasis, TNM staging and HER2 over expression (P < 0.01, P < 0.05 and P < 0.01, respectively), and inversely correlated with PR expression (P < 0.05). The patients' relapse-free survival was shorter in TGFbeta1-positive group than in TGFbeta1 negative group (P < 0.05). Both the expression of MMP-2 and MMP-9 were positively correlated with CD147 expression; and both the expression of TGFbeta1 and TGFbetaRI were positively correlated with CD147 expression (P < 0.01 and P < 0.05, respectively).

CONCLUSIONThe expression of CD147 is considered closely correlated with tumor invasion, metastasis and prognosis in breast cancer, and has also a close correlation with MMP-2, MMP-9, TGFbeta1 and TGFbetaRI expression.

Adult ; Aged ; Basigin ; metabolism ; Breast Neoplasms ; metabolism ; pathology ; Carcinoma, Ductal, Breast ; metabolism ; pathology ; Female ; Fibroadenoma ; metabolism ; pathology ; Follow-Up Studies ; Humans ; Lymphatic Metastasis ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Middle Aged ; Neoplasm Invasiveness ; Neoplasm Staging ; Protein-Serine-Threonine Kinases ; metabolism ; Receptor, ErbB-2 ; metabolism ; Receptors, Progesterone ; metabolism ; Receptors, Transforming Growth Factor beta ; metabolism ; Survival Rate ; Transforming Growth Factor beta1 ; metabolism ; Young Adult

Fruit ripening is associated with a number of physiological and biochemical changes. They include degradation of chlorophyll, synthesis of flavor compounds, carotenoid biosynthesis, conversion of starch to sugars, cell wall solublisation and fruit softening. These changes are brought about by the expression of specific genes. People are interested in the molecular mechanism involved in the regulation of gene transcription during fruit ripening. Many fruit-specific promoters such as PG, E4, E8, and 2A11 have been characterized and shown to direct ripening-specific expression of reporter genes. AGPase plays the key role in catalyzing the biosynthesis of starch in plants. It is a heterotetrameric enzyme with two small subunits and two large subunits, which are encoded by different genes. In higher plants, small subunits are highly conserved among plant species and expressed in all tissues. And the large subunits are present at multiple isoforms and expressed in a tissue-specific pattern. In fruits, the expression pattern of the large subunits varies with plant species. That made it important to study the transcriptional regulation of the large subunits of AGPase in different plant species. Northern-blot analysis indicates in watermelon, an isoform of the large subunits Wml1 expressed specifically in fruits, not in leaves. The 5' flanking region of Wml1, which covers 1573bp, has been isolated through the method of uneven PCR. And transient expression assay has shown that the 1573bp (named WSP) can direct fruit-specific expression of GUS gene. Our goal in this study was to scan the promoter region for main regulatory regions involved in fruit-specific expression. A chimaeric gene was constructed containing the WSP promoter, the beta-glucuronidase (GUS) structural sequence as a reporter gene and the nopaline synthase polyadenylation site (NOS-ter). The plasmid pSPA was digested with Hind III + Hinc II and promoter fragment of 1573bp (from 180bp to 1752bp) was cut out and cloned into Sma I sites of pBluescript SK(-), to produce pBSPA-16. The same insert was then cut out with Hind III + BamH I, and ligated with transient expression vector pBI426 digested by HindIII + Bgl II to produce pISPA-16. Three 5'-end deletions of the promoter were obtained and fused to GUS gene in plant transient expression vector pBI426: the 1201bp fragment (from 551bp to 1752bp) was generated by digestion of pBSPA-16 with BamH I + SnaB I, the 898bp fragment (from 854bp to 1752bp) by BamH I + EcoRV. Both fragments were ligated with pBluescript SK(-) digested by BamH I + Sma I, to produce pBSPA-12 and pBS-PA-9. The inserts were cut out with HindmIII + BamH I and ligated with pBI426 digested by Hind III + Bgl II, to produce pISPA-12 and pISPA-9. The 795bp fragment (from 957bp to 1752bp) was generated by digestion of pSPA with Hinc II + EcoR I, promoter fragment was cut out and cloned into Sma I sites of pBluescript SK(-), to produce pBSPA-8. The same insert were cut out with Hind III + BamH I, and ligated with transient expression vector pBI426 digested by Hind III + Bgl II. The 1573bp fragment and three 5'-end deletions were delivered into watermelon leaf, stem, flower and fruit of different development stages (5, 10, 20 days after pollination) via particle bombardment using a biolistic PDS-1000/He particle gun. Bombardment parameters were as follows: a helium pressure of 1200 psi, vacuum of 91432.23Pa, 7 cm between the stopping screen and the plate. Histochemical assay were done on all the tissues bombarded after incubation for 2 days. The 1573bp fragment had the strongest promoter activity, and can induce GUS expression in fruits of 5 and 20 days after anthesis and flowers, but not in fruits of 10 days after anthesis, leaves and stems. Fragments of 1201bp and 898bp can induce GUS expression only in fruits of 20 days after anthesis, and with lower expression levels than 1573bp. Fragment of 795bp was not able to direct GUS expression in any of the tissues bombarded (data not shown). It can be concluded that of the 1573bp, 1201 bp, 898bp Wml1 5'flanking regions include the necessary information directing fruit-specific expression. Deletion from 180bp to 551bp doesn't affect the fruit-specificity of the promoter, but lowered the expression level. There may be some cis-acting elements located in this region, which can enhance external gene expression in later stages of fruit development. Deletion from 854bp and 958bp led to loss of GUS expression. This region includes the necessary information needed for gene expression as well as the regulatory elements for fruit-specific transcription.
Citrullus ; genetics ; Fruit ; genetics ; Gene Expression Regulation, Plant ; genetics ; Promoter Regions, Genetic ; genetics ; Regulatory Sequences, Nucleic Acid ; genetics ; physiology

Objective To investigate the basic distribution of endemic areas in the type of drinking water arsenism and in Xiantao City,Hubei Province,and to offer a scientific basis for control and prevention.Methods According to"the Chinese Scheme of Implementing Surveillance of Distribution of Endemic Arsenism",considering with the special geography feature of Xiantao,both sampling and overall survey were used in 7 towns chosen.The water arsenic content was determined by half quantitative fast reagent-box method.We began to search for clues and patients according to the endemic areas and the families with high arsenic wells.Results High endemic arsenic water sources were distributed in 7 the towns(districts or farms).In 81 villages of Xiantao City,35 villages had drinking water arsenic content exceeding 0.05 mg/L,accounting for 43.21%(35/81).In 4020 screened wells,269 had arsenic content higher than the national standard,the detective rate of high arsenic wells(more than 0.05 mg/L)was 6.69%(269/4020),with the highest rate in Shahu Seed Plant being 13.56%(115/848).The population exposed to high arsenic was 1091,in a rate of 5.75%(1091/18 975),in which 281 children were exposed in a rate of 5.82%(281/4826).In Shahu Seed Plant,467 people including 129 children were exposed to high arsenic, accounting for 13.26%(467/3522)and 12.91%(129/999),respectively.Conclusions High arsenic sources widely exist in Xiantao City,especially in Shahu Seed Plant,where arsenic content,the exposed rate of population and children are high.Therefore,prevention and control should be carried out in the southeast as soon as possible,as well as in other places where situation is less serious.