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

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

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 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 the relationship between the extracellular matrix (ECM) and neoplastic progression in hamster with tongue cancer.

METHODSForty-eight specimens of hamster tongue cancer were divided into control group (n=6) and experimental group (n=42). The pathological grade of the specimens was assessed (including 3 stages, namely atypical hyperplasia, carcinoma in situ and early invasive carcinoma). The sections of the tongue were stained with Masson and aldehyde-fuchsin (AF) staining for microscopic observation of the elastic fiber and collagen fiber changes.

RESULTSWithin the connective tissue cores (CTC) of the papillae in the control group was a framework of numerous and fine Gomrori's aldehyde fuchsin-positive elastic fibers. But in the stages of dysplasia and carcinoma in situ, these elastic fibers decreased and further diminished in the CTC in early invasive carcinoma. In dysplasia and carcinoma in situ stages, most of the elastic fibers collapsed with scattered elastic fibers, and the elastic fibers decreased significantly in early invasive carcinoma. The control group showed a significantly greater number of elastic fibers in the experimental group. The collagen fiber was obviously increased and irregularly arranged in dysplasia and carcinoma in situ stage; in early invasive carcinoma, the collagen fibers became thicker with deposition in the lamina propria.

CONCLUSIONAn excessive deposition of collagen fiber and reduction of the elastic fibers is an important factor contributing to the development of tongue carcinoma in hamsters.

Animals ; Carcinoma ; pathology ; Collagen ; metabolism ; Connective Tissue ; pathology ; Cricetinae ; Elastic Tissue ; pathology ; Extracellular Matrix ; pathology ; Neoplasms, Experimental ; pathology ; Tongue Neoplasms ; pathology

OBJECTIVETo detect the expression and the CpG island methylation status of tumor suppressor gene p15 after exposure to 1,4-benzoquinone (1,4-BQ) in primary cultivated C57BL/6J mouse bone marrow cells in vitro.

METHODSThe mouse bone marrow cells were isolated in vitro. The effect of 0, 0.1, 1, 5, 10, 20, and 40 µmol/L 1,4-BQ on cell viability (CKK-8) was detected. 0, 0.1, 1, 10 µmol/L 1,4-BQ were used to intoxicate the mouse bone marrow cells for 24 h; Real-time PCR was employed to analyze the mRNA expression level of p15; The bisulfite sequencing PCR (BSP) was used to look into the methylation status of CpG islands in p15 promoter region.

RESULTS1,4-BQ exhibited dose-dependent toxicity to mouse bone marrow cells, and the LC(50) was 8.3 µmol/L (95%CI: 4.6 - 10.6 µmol/L). The mRNA expression of p15 in 10 µmol/L group was only equivalent to 43% of control group. Compared with control group, the decrease of p15 mRNA expression in1 and 10 µmol/L concentration were obvious, and the differences had statistical significance (P < 0.05 or P < 0.01). BSP sequencing results were same between the exposure groups and control group, the 56 CpG sites on CpG islands remained in the state of unmethylated.

CONCLUSIONmRNA expression of p15 gene decreases after exposure to 1,4-BQ, but the CpG islands methylation status in promoter is not affected, suggesting that methylation does not participate in 1,4-BQ-mediated p15 gene expression decrease, other effect mechanisms still need to be investigated.

Animals ; Base Sequence ; Benzoquinones ; toxicity ; Bone Marrow Cells ; metabolism ; Cells, Cultured ; CpG Islands ; Cyclin-Dependent Kinase Inhibitor p15 ; genetics ; DNA Methylation ; Environmental Exposure ; Mice ; Mice, Inbred C57BL ; Promoter Regions, Genetic ; RNA, Messenger ; genetics

OBJECTIVETo investigate potential mutations and clinical features of 9 unrelated patients with inherited coagulation factor VII (FVII) deficiency.

METHODSClinical diagnosis was validated by assaying of coagulation parameters including prothrombin time, activated partial thromboplastin time, FVII activity and specific antigens. All exons, exon-intron boundaries, and 5' and 3' untranslated regions of F7 genes were amplified with PCR. Potential mutations were detected by direct sequencing of purified PCR products. Suspected mutations were confirmed by sequencing of the opposite strand.

RESULTSAll probands have featured prolonged prothrombin time, with FVII activity ranging between 2.0% to 6.0%. The titers of FVII antigen were significantly reduced in 7 probands. Eight mutations, including 6 missense mutations, 1 deletion and 1 insertion, were identified, among which 3 (Gln100Leu, Ser269Pro and g.11520_11521insT) were not described previously. Six mutations have located in the protease domain. All mutations were inherited, and consanguineous marriages were reported in 5 families. Mutations g.27_28delCT, Cys329Gly, Arg304Trp and His348Gln have been identified in unrelated families. There was a lack of correlation between the mutations and their clinical features. Two individuals with homozygous His348Gln mutations and 1 individual with homozygous Arg304Trp mutation were only mildly affected or asymptomatic. Two patients, who have respectively carried homozygous and heterozygous deletions of g.27_28delCT, were moderately affected and asymptomatic. In 4 patients carrying double heterozygous mutations, 1 (Ser269Pro and Cys329Gly) was asymptomatic, 2 (Arg304Trp and Cys329Gly, Arg277Cys and g.11520_11521insT, respectively) had a mild bleeding tendency, whilst 1 (Gln100Leu and His348Gln) has a moderate bleeding diathesis.

CONCLUSIONThere seem to be hotspots of F7 gene mutations in ethnic Han Chinese populations. And there is a lack of correlation between particular types of mutations and clinical phenotypes.

Adolescent ; Adult ; Aged ; Base Sequence ; Blood Coagulation Disorders, Inherited ; genetics ; Child ; Factor VII ; genetics ; Factor VII Deficiency ; genetics ; Female ; Heterozygote ; Homozygote ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Mutation ; Young Adult