Female ; Humans ; Infant, Newborn ; Maple Syrup Urine Disease ; diagnosis ; therapy

The objective of the present study was to establish a method based on principal component analysis (PCA) for the study of transdermal delivery of Chinese medicinal formulae, and to choose the best penetration enhancers for Qingfei Xiaocuo gel depend on this method. Using improved Franz type diffusion cell and excised rat skin in vitro as transdermal barrier, the receptive solution fingerprint was established by HPLC, harvesting the areas of the common peaks in the fingerprint, then the total factor scores of the concentrations at different times were calculated using PCA and were employed instead of the concentrations to compute the cumulative amounts (Q12) and enhancement ratio (ER), the latter of which were considered as the indexes for optimizing penetration enhancers. Compare to the control group, the ER of the other groups increased significantly and furthermore, 2.5% azone with 2.5% menthol manifested the best effect. PCA represent most information in the receptive solution, the method above could choose the best penetration enhancers, it could be a reference for the study of transdermal delivery of Chinese medicinal formulae.
Administration, Cutaneous ; Animals ; Drugs, Chinese Herbal ; analysis ; pharmacokinetics ; Gels ; In Vitro Techniques ; Male ; Medicine, Chinese Traditional ; Mice ; Principal Component Analysis ; Skin ; metabolism

In comparison with traditional organic dyes, semiconductor quantum dots ( QDs) feature a series of superior luminescence properties, including narrow and symmetric emission, broad excitation and strong absorption, excellent photobleaching-resistance, and good water solubility. The addition of dopants can endows QDs with extra new properties, e. g. , further increase the Stokes shift for avoiding self-quenching. Mn-doped ZnS QDs are a very representative example of doped QDs. No harmful elements such as Cd and Hg are involved in such type of bio-friendly QDs. Besides, the Mn2+dopant further adds QDs with excellent room temperature phosphorescence. Phosphorescent detection can effectively eliminate the interference of biological background fluorescence, thus Mn-doped ZnS QDs can be widely used in phosphorescent bioanalysis. In this paper, the recent progress of room temperature phosphorescence analysis with Mn-doped ZnS QDs was reviewed. The emphasis was placed on the several stimulative sensing design strategies, including the luminescence mechanism, ion probes, detection of small molecules and biomacromolecules.

Objective To observe the protein and gene expression of immunoglobulin binding protein (BiP) in the femur of fluoride-treated rats, and preliminarily study the possible role of endoplasmic reticulum stress in the pathogenesis of skeletal fluorosis. Methods Sixty Wistar rats were divided into 4 groups according to body weight, n =15. The control and low-calcium groups were fed with normal diet(0.79％ calcium) and low-calcium diet(0.063％ calcium), respectively, and both drank tap water(fluoride concentrations ＜ 1 mg/L). High-fluoride and coexpesure to low-calcium groups were fed with conventional feed (0.79％ calcium) and low-calcium diet (0.063％ calcium), respectively, and both drank tap water containing sodium fluoride (sodium fluoride concentration of 221 mg/L). During experimental period, rats were measured body weight once a week with a stand diet and water available ad libitum. The experiment lasted for 12 weeks. The immunohistochemical and reverse transcription polymerase chain reaction(RT-PCR) techniques were used to detect the protein and gene expression of BiP in the femur of fluoride-treated rats and control subjects. Results The bone mineral contents of high fluoride, lowcalcium and coexposure groups[(0.131 ± 0019), (0.097 ± 0.011 ), (0.083 ± 0.007)g/cm] were lower than those of the control group[(0.159 ± 0.029)g/cm, all P ＜ 0.05]; the bone mineral density of low calcium and coexpesure to fluoride group[(0.243 ± 0.018), (0.223 ± 0.022)g/cm2] was lower than that of the control group[(0.296 ± 0.046)g/cm2, all P ＜ 0.05]. The immunohistochemical staining showed that the anti-BiP antibody positive osteoblasts were significantly increased in the low calcium diet and coexposure to fluoride groups than that in the control, and coexposure to fluoride elevated the positive cells than that in only low calcium diet group. The mRNA expression of osteopontin(OPN) and osteocalcin(OCN) in coexposure to fluoride with low-calcium group(1.36 ± 0.20, 1.31 ±0.11 ) was higher than that of the control groups (0.82 ± 0.16, 0.85 ± 0.15, all P ＜ 0.05) ; moreover, OPN expression significantly increased in this group than that of the only high fluoride group (0.97 ± 0.29, P ＜ 0.05). The mRNA expression of BiP in the low calcium and coposure to fluoride group (1.38 ± 0.24,1.35 ± 0.12) was significantly higher than that of the control group ( 1.14 ± 0.06, all P ＜ 0.05 ). Conclusions Higher fluoride or coexposure to low calcium diet stimulates the gene and protein expression in rat femur BiP, indicating that varying degrees of endoplasmic reticulum stress is likely involved in the pathogenesis of rat skeletal fluorosis.

To study effect of vitrification state of protective solutions on recovery of red blood cells after lyophilization, four protective solutions composed of isotonic buffers containing 7% DMSO (v/v) and 20%, 30%, 40% or 50% polyvinylpyrrolidone (PVP) (w/v) were adopted. Vitrification state of protective solutions was examined first when white ice crystal appeared in any protective solution during freezing or thawing, if the used solution was not a vitrification solution. Red blood cells were lyophilized in MINILYO45 freeze-dryer after washing, mixing with protective solutions and prefreezing. After lyophilization, the samples were quickly rehydrated by 37 degrees C rehydration solution. The results showed that in vitrification and devitrification experiments, white ice crystal appeared in solution of 20% PVP + 7% DMSO and 30% PVP + 7% DMSO during freezing and thawing; vitrification appeared in solution of 40% PVP + 7% DMSO during freezing, but devitrification appeared during thawing; vitrification appeared in solution of 50% PVP + 7% DMSO during freezing and thawing. After rehydration, the recoveries of red blood cells and hemoglobin in 40% PVP + 7% DMSO group were (81.36 +/- 14.94)% and (77.54 +/- 12.86)%, which were significantly higher than that in 20% PVP + 7% DMSO, 30% PVP + 7% DMSO and 50% PVP + 7% DMSO groups (P < 0.01). The concentration of free hemoglobin in 40% PVP + 7% DMSO group was also significantly lower than that in other three groups (P < 0.01). With increase of PVP concentration in protective solutions, vitrification state and protective effect of these solutions also increased; when concentration of PVP in protective solution was 40% though it was not a vitrification solution, the effect of lyophilization was the best; but when concentration of PVP further increased to 50%, though it was a vitrification solution, the effect decreased. It is concluded that excessive vitrification state could not benefit lyophilization of red blood cells.
Cryoprotective Agents ; pharmacology ; Dimethyl Sulfoxide ; pharmacology ; Dose-Response Relationship, Drug ; Erythrocytes ; cytology ; drug effects ; ultrastructure ; Freeze Drying ; methods ; Humans ; Microscopy, Electron ; Povidone ; pharmacology

Objective Comparative and statistical analysis the HPV infection rate between fresh tissue and Paraffin-embedded Specimens of esophageal squamous cell carcinoma,and comparative the testing results with others regions.Methods Extracted the total DNA from the novel fresh tissue and Paraffinembedded Specimens; Detected the DNA by PCR with universal primer and Detected the HPV type with human papilloma virus nucleic acid amplification-based typing detection reagent kit (Hybribio) ; Compared the statistical result from the different specimens; analyzed the result between different region.Results HPV infection rate of fresh tissue is 82.6％ with HPV16 (34.8％ ) and HPV18 ( 34.8％ ),and paraffinembedded specimens is 78.2％ with HPV16 ( 30.4％ ) and HPV18 ( 17.4％ ).Conclusion The results provides the first evidence that there wasn' t noticeable difference between HPV infection rate of the two specimens.So broader specimen source could be used for HPV testing.

Objective To characterize genotypic resistance within HBV RT region in chronic hepatitis B (CHB) patients with nucleos(t) ide analogue (NA) treatment.Methods Serum samples of 229 CHB patients with NA treatment were obtained.Full-length HBV RT sequences were amplified,sequenced and analyzed,on the following NA resistant (NAr) mutations belonging to different NAr pathways.Results Among 229 HBV isolates,14.41％ (33/229) and 85.59％ (196/229) were genotype B and C,respectively;and the patients with HBV genotype C may be more susceptible to develope resistant mutations than patients with HBV genotype B (x2 =2.95,P ＜ 0.05).NAr mutations were detected in 63 CHB patients.Mutations were not found at rtI169,rtT184,rtA194 or rtS202.RtM204 mutations were detected at the highest frequency among 63 mutants (40/63,63.49％) and found to display 11 combination mutation patterns,in which rtM204I were associated with rtL80I/V and rtL180M,and rtM204V were associated with rtL180M,respectively.Conclusions There are complicated mutation patterns in the HBV RT region for chronic hepatitis B (CHB) patients with nucleos(t) ide analogue (NA) treatment.RtM204V/I mutation was the highest.

OBJECTIVETo investigate whether dendritic cells pulsed with whole tumor lysates (WTL) could in vitro elicit antitumor T cell responses in patients with non-small-cell lung cancer (NSCLC).

METHODSMonocyte-derived immature DCs (imDCs) generated in the presence of human recombinant granulocyte-macrophage colony stimulating factor and interleukin-4 from peripheral blood mononuclear cell of NSCLC patients, and then were induced to mature by pulsing autologous WTL (DCs/WTL) or by the addition of TNF-alpha(TNF/DCs). FACS and MLR assay were used to monitor their phenotypic changes and capacity to stimulate allogeneic and autologous T cell proliferation. DCs/WTL activated with TNF-alpha (* DCs/WTL) were cocultured in vitro with autologous T cells for eliciting antitumor CTLs. T cell mediated antitumor responses were measured by IFN-gamma enzyme-linked immunospot (ELISPOT) assay for WTL-specific IFN-gamma releasing T cells and by lactate dehydrogenase release (LDH) assay for lysis of autologous tumor cells, respectively.

RESULTSWhen monocytes-derived imDCs from the patients with NSCLC (n = 10) were pulsed with autologous WTL for a day at 30 microg total protein of WTL per 10(6) DCs/ml, this led to up-regulation of CD1a, CD83 and CD86 as well as HLA-DR, and also led to marked stimulation of allogeneic T cell proliferating activity, which was comparable to that of TNF/DCs. However, their capacity of stimulating autologous T cell proliferation in vitro was significantly more potent than those of TNF/DCs (P < 0.05). The numbers of WTL-specific IFN-gamma releasing T cells in 1/3 cultures after one week exposure to * DCs/WTL was increased significantly compared with those pulsing with TNF/DCs plus IL-2 or IL-2 alone (P = 0.05). T cells derived by priming of non-adherent PBMCs with * DCs/WTL after 14 days in vitro stimulation were significantly more responsive to autologous tumor cells compared with LAK (n = 3, P < 0.05), but its cytotoxicity against K562 cells was also comparable to LAK cells.

CONCLUSIONMonocyte-derived DCs from NSCLC patients could serve as functional APC. The * DCs/WTL may effectively elicit T cell-mediated antitumor response in vitro and enhance NK killing activity.

Antigens, CD1 ; metabolism ; Carcinoma, Non-Small-Cell Lung ; immunology ; Cell Culture Techniques ; Cytotoxicity, Immunologic ; Dendritic Cells ; immunology ; HLA-DR Antigens ; metabolism ; Humans ; Interferon-gamma ; secretion ; K562 Cells ; Killer Cells, Lymphokine-Activated ; immunology ; Leukocytes, Mononuclear ; immunology ; pathology ; Lung Neoplasms ; immunology ; Lymphocyte Culture Test, Mixed ; T-Lymphocytes, Cytotoxic ; immunology ; pathology ; Tumor Necrosis Factor-alpha ; pharmacology

To study the dynamic changes of ultrastructure of erythrocytes in prolonged preservation of blood with preservative fluid containing superoxide dismutase (SOD), the whole blood samples were preserved at 4 degrees C in SOD-containing solution, the morphologic changes of erythrocyte were dynamically ob served by transmission microscopy after preservation for 42, 75 and 85 days, an d the blood samples preserved in GMA solution served as control. Three variance was applied to analyze the data with SAS software. The results showed that the metamorphotic rates of erythrocyte preserved in SOD-containing solution for 42, 75 and 85 days were lower than those of erythrocytes preserved in GMA solution. Most of metamorphotic rates of erythrocyte preserved in SOD-containing solution for 42, 75 and 85 days were correspond to those of erythrocytes preserved in GMA solution for 42 days, or even lower. It is concluded that SOD-containing preservative fluid might help to maintain the normal morphology of erythrocytes in prolonged preservation at 4 degrees C.
Blood Preservation ; Erythrocyte Deformability ; Erythrocytes ; ultrastructure ; Humans ; Microscopy, Electron ; Superoxide Dismutase ; pharmacology ; Time Factors

BACKGROUNDMyopia is a common disorder and the incidence has increased yearly, but its pathogenesis remains unclear. The aim of this study was to investigate the possible role of hepatocyte growth factor (HGF) and its receptor c-Met in the development of lens-induced myopia in guinea pigs.

METHODSSixty one-week-old guinea pigs were chosen. The right eyes were treated with -10.0 diopters (D) lenses as the lens-induced myopia group; the left eyes remained untreated as the control group. Six weeks later, refractive status and axial length were determined by streak retinoscopy and A-scan ultrasonography, respectively. The guinea pigs were killed and both eyes collected. Morphological changes were observed by hematoxylin and eosin staining. The expression levels of HGF, c-Met, and matrix metalloproteinase 2 (MMP-2) mRNA and protein in the posterior sclera were analyzed by RT-PCR and Western blotting, respectively.

RESULTSThe lens-induced myopia group became myopic with a significant increase in axial length and a significant decrease in refraction. Compared with the control group, the posterior retina and sclera were thinner in the lens-induced myopia group. The expression levels of HGF and MMP-2 mRNA and protein and of phosphorylated c-Met protein were significantly higher in the posterior sclera of the lens-induced myopia group than in the control group (all P < 0.05). In the lens-induced myopia group, the expression level of MMP-2 in the posterior sclera positively correlated with the expression level of HGF (r = 0.902, P < 0.05) and phosphorylated c-Met (r = 0.885, P < 0.05).

CONCLUSIONHGF/c-Met might play a role in the development of lens-induced myopia in guinea pigs by upregulating the expression of MMP-2.

Animals ; Guinea Pigs ; Hepatocyte Growth Factor ; metabolism ; Myopia ; etiology ; metabolism ; Proto-Oncogene Proteins c-met ; metabolism