Accidents, Occupational ; statistics & numerical data ; China ; epidemiology ; Humans

A simple, sensitive and reproducible high-performance liquid chromatography (HPLC) method has been validated for determining concentrations of glutamate, glycine, and alanine in human plasma. Proteins in plasma were precipitated with perchloric acid, followed by derivatization with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC). Simultaneous analysis of glutamate, glycine, and alanine is achieved using reversed-phase HPLC conditions and ultraviolet detection. Excellent linearity was observed for these three amino acids over their concentration ranges with correlation coefficients (r)>0.999. The intra- and inter-day precision were below 10%. This method utilizes quality control samples and demonstrates excellent plasma recovery and accuracy. The developed method has been successfully applied to measure plasma glutamate, glycine, and alanine in twenty volunteers.
Alanine ; Amino Acids ; Aminoquinolines ; Carbamates ; Chromatography, High Pressure Liquid ; Chromatography, Liquid ; Glutamic Acid ; Glycine ; Humans ; Perchloric Acid ; Plasma ; Proteins ; Quality Control

Soxhlet extraction is a common method of sample preparation. However, there has been no discussion about the efficiency of Soxhlet extraction from different batches and the factors that cause content fluctuation. In this study, Panax ginseng was selected as a model sample. Soxhlet extraction by means of a water bath, which has always been neglected, was identified as a novel key factor in the poor repeat-ability in different batches of Soxhlet extraction, as it can affect the siphon times and reflux time, which have been positively correlated with the ginsenoside contents. By substituting round bottom flasks in the same column, the relative standard deviation of the most fluctuated compound, ginsenoside Rb1, was decreased from 24.6% to 5.02%. Scanning electron microscopy analysis confirmed that the breakdown of the surface of the ginseng powder in the Soxhlet extraction led to a better dissolution of ginsenosides, indicating that chloroform may promote the extraction of ginsenosides by disrupting the cell structure. Moreover, 70% methanol was regarded as the better solvent for extracting the ginsenosides. Overall, this work offers a practical and effective protocol for improving the accuracy and repeatability of Soxhlet extraction methodology for ginsenosides and other analytes.

BACKGROUND AND PURPOSE: Five single-nucleotide polymorphisms (SNPs) (rs4379368, rs10504861, rs10915437, rs12134493 and rs13208321) were recently identified in a Western population with migraine. These migraine-associated SNPs have not been evaluated in a Han Chinese population. This study investigated the associations of specific SNPs with migraine in a Han population. METHODS: This was a case-control study of Han Chinese residing in Fujian Province. Polymerase chain reaction—restriction-fragment-length polymorphism analysis and direct sequencing were used to characterize the relationships of SNPs in a control group of 200 subjects and in a migraine group of 201 patients. RESULTS: The frequencies of the five SNPs did not differ between patients with migraine and healthy non migraine controls. However, subgroup analysis indicated certain SNPs were more strongly associated with migraine with aura or migraine without aura than with controls. The CT genotype of rs4379368 was more common in migraine patients with aura (75%) than in migraine patients without aura (47.9%) and controls (48.5%) (p<0.05), and the TT genotype of rs10504861 was more common in migraine patients with aura than in controls (8.3% vs. 0.5%) (p<0.05). Meanwhile, the CC genotype of rs12134493 was less common in migraine patients without aura than in controls (80.6% vs. 88%) (p<0.05). CONCLUSIONS: Our findings suggest that the rs4379368 and rs10504861 SNPs are markers for susceptibility to migraine with aura and that rs12134493 is a marker for the risk of migraine without aura in this Han population. Future studies should further explore if these associations vary by ethnicity.
Asian Continental Ancestry Group* ; Case-Control Studies ; Epilepsy ; Genotype ; Humans ; Migraine Disorders* ; Migraine with Aura ; Migraine without Aura ; Polymorphism, Single Nucleotide

OBJECTIVETo explore the quality of life (QOL) outcome in patients with allergic rhinitis (AR).

METHODSA prospective trial was conducted to survey the QOL status of 101 AR patients, in contrast to that of 121 healthy individuals and 97 chronic pharyngitis (CP) patients by generic questionnaire medical outcomes study short-form 36-items health survey (SF-36), and to survey the most troublesome problems of AR patients by disease-specific questionnaire rhinoconjunctivitis quality of life questionnaire (RQLQ). The correlation between SF-36 and RQLQ had also been analyzed. All the results were analyzed statistically.

RESULTSBy the assessment of SF-36, the scores of 3 domains (x ± s, the same as follow, the scores were 78.02 ± 18.37, 56.13 ± 17.49, 78.81 ± 16.47, respectively) of AR patients were less than those (84.00 ± 18.36, 74.69 ± 14.13, 83.78 ± 14.31) of healthy individuals (P < 0.05), and the scores of 7 domains (the scores were: 91.78 ± 11.78, 79.16 ± 30.23, 78.02 ± 18.37, 56.13 ± 17.49, 78.81 ± 16.47, 67.66 ± 39.57, 68.78 ± 13.65, respectively) of AR patients were similar with those (94.12 ± 6.88, 80.67 ± 32.38, 73.57 ± 17.96, 59.73 ± 16.58, 80.41 ± 17.01, 63.58 ± 39.99, 66.43 ± 13.71) of CP patients (P > 0.05). By the assessment of RQLQ, in AR patients, both the nasal symptoms and the practical problems got the highest scores (the scores were 2.70 ± 1.29, 2.53 ± 1.37 respectively). According to the assessment of the correlation between SF-36 and RQLQ, the correlation was weak (r = -0.199 ∼ -0.526, P < 0.05).

CONCLUSIONSThe QOL of AR patients decreased compared with that of healthy individuals, but similar with that of CP patients. The most troublesome problems in AR patients were nasal symptoms and the practical problems. Both SF-36 and RQLQ were suitable for assessing the health status of AR patients. SF-36 and RQLQ each covered a different part of the QOL of AR patients, and the combination of the two questionnaires could improve the QOL measurement.

Adolescent ; Adult ; Female ; Humans ; Male ; Middle Aged ; Prospective Studies ; Quality of Life ; Rhinitis, Allergic, Perennial ; epidemiology ; Surveys and Questionnaires ; Young Adult

To investigate the expression and localization of various functional domains of ORF1 polyprotein and ORF3 protein of hepatitis E virus in host cells, the coding sequences of the various functional domains (RdRp, HEL, MET, PLP, X) of ORF1 were separately cloned into pcDNA3. 1-GFP vectors for constructing the recombinant plasmids which were verified by enzyme digestion and sequencing. The exact expression of the fusion proteins were detected by Western Blot, and the distribution and localization were observed by the laser scanning confocal microscope(LSCM). In huh7 cells, GFP-RdRp proteins were found mainly in the nuclei, GFP-HEL proteins were distributed vesicularly around the nucleus, GFP-MET proteins were distributed granularly both in the nuclei and the cytoplasm, GFP-PLP proteins had polar distribution around the nucleus, and unknown GFP-X proteins were distributed uniformly both in the nuclei and the cytoplasm. Different localization of these proteins verified the previous data obtained from in vitro studies, providing a support for further research on the biological functions of various proteins coded by HEV genome.
Blotting, Western ; Cells, Cultured ; Hepatitis E virus ; genetics ; Humans ; Open Reading Frames ; Viral Proteins ; genetics ; physiology

OBJECTIVETo study the impact of pulmonary fibrosis on erectile function in rats and its mechanism.

METHODSForty 12-week-old healthy male SD rats were randomly divided into Groups A (4-week pulmonary fibrosis), B (6-week pulmonary fibrosis), C (4-week control, and D (6-week control). The models of pulmonary fibrosis were established by injection of bleomycin at 5 mg/kg in the trachea, while the controls were injected with normal saline only. At 4 and 6 weeks, all the rats were subjected to determination of the serum testosterone (T) level, arterial blood gas analysis, measurement of intracavernous pressure/mean arterial pressure (ICP/MAP), and examination of NOS activity and cGMP content. The mRNA expressions of eNOS, iNOS and nNOS in the corpus cavernosum penis were detected by real-time PCR, and that of eNOS analyzed by Western blot.

RESULTSThe 3 V and 5 V of the ICP/mapx100 in Group C were 16.37 +/- 2.19 and 27.19 +/- 3.18, significantly lower than 30.78 +/- 2.66 and 50.09 +/- 6.97 in Group A (P < 0.05); those in Group D were 10.17 +/- 1.31 and 17.40 +/- 1.74, significantly lower than 31.45 +/- 3.07 and 51.23 +/- 7.23 in Group B (P < 0.05), and so were they in Group D than in C (P < 0.05). PaO2 was significantly lower in Group C than in A ([75.50 +/- 13.87] mmHg vs [103.80 +/- 6.88] mmHg, P < 0.05) , and so was it in Group D than in B ( [83.60 +/- 5.50] mmHg vs [102.70 +/- 5.77] mmHg, P < 0.05). Group C showed a significantly increased serum T level as compared with A ([391.1 +/- 264.7] ng/dl vs [175.9 +/- 53.0] ng/dl, P < 0.05), so did Group D ([745.4 +/- 408.8] ng/dl) versus Group B ([177.8 +/- 52.3] ng/dl) and C (P < 0.05). NOS activity and cGMP content in the corpus cavernosum significantly decreased in Group C ([1.50 +/- 0.14] U/mg prot and [35.69 +/- 3.64] pmol/mg) compared with A ([2.66 +/- 0.39] U/mg prot and [51.10 +/- 7.22] pmol/mg) (P < 0.05), so did they in D ([1.40 +/- 0.20] U/mg prot and [34.55 +/- 4.30] pmol/mg) versus B ([2.75 +/- 0.36] U/mg prot and [52.15 +/- 6.86] pmol/mg) (P < 0.05), but neither showed any significant difference between Groups D and C (P > 0.05). The expression of the eNOS protein was significantly lower in Group C than in A (0.79 +/- 0.01 vs 0.87 +/- 0.01, P < 0.05), so was it in D than in B and C (0.71 +/- 0.02 vs 0.88 +/- 0.01 and 0.79 +/- 0.01, P < 0.05). The expression of eNOS mRNA was significantly higher in Group C than in A (4.46 +/- 0.92 vs 2.61 +/- 0.68, P < 0.05), but did not show any significant difference between D and B (2.79 +/- 0.60 vs 2.69 +/- 0.65, P > 0.05), nor did the expressions of nNOS mRNA and iNOS mRNA between the pulmonary fibrosis groups and the controls (P > 0.05).

CONCLUSIONPulmonary fibrosis may induce erectile dysfunction by suppressing the expression of the eNOS protein and reducing NOS activity and cGMP content in the corpus cavernosum penis of rats.

Animals ; Erectile Dysfunction ; etiology ; metabolism ; Male ; Nitric Oxide Synthase ; metabolism ; Penis ; metabolism ; Pulmonary Fibrosis ; complications ; metabolism ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo measure lutein, zeaxanthin and β-carotene level in foods commonly consumed in Beijing, and compare the content difference between raw and cooked food.

METHODSForty-six commonly consumed foods of 8 classes were collected in Haidian district of Beijing from September to October in 2009. A high performance liquid chromatography method was used to determine the content of lutein, zeaxanthin and β-carotene in both raw and cooked samples.

RESULTSLutein was abundant in cucurbitaceous and solanaceous, allium and nuts, especially in Chinese chive (18 226.9 µg/100 g) and pumpkin (13 265.2 µg/100 g). Major sources of zeaxanthin included round pumpkin, green garlic shoot, corn and eggs, whose level of zeaxanthin were 444.6, 283.5, 279.7, 118.6 - 377.9 µg/100 g, respectively. Zeaxanthin level of those cooked foods changed to 483.9, 239.3, 279.1, 149.5 - 594.7 µg/100 g, respectively. The zeaxanthin level of cooked Chinese chive reached 1081.2 µg/100 g, while we did not detect any zeaxanthin in raw Chinese chive. β-carotene was present in a wide variety of vegetables and fruits. Carrot (17 234.3 µg/100 g) was a good source of β-carotene, while its level in cooked carrot was 17 013.5 µg/100 g.

CONCLUSIONConsuming the proper kinds of foods and changing the method of food processing were beneficial to increase the intake of lutein, zeaxanthin and β-carotene.

China ; Cooking ; Food ; Food Analysis ; Lutein ; analysis ; Xanthophylls ; analysis ; Zeaxanthins ; beta Carotene ; analysis

AIM:To investigate the mechanism of the radiosensitizing effect of maximum non-cytotoxic doses of tetrandrine (Tet) on nasopharyngeal carcinoma cell lines CNE1 and CNE2.METHODS:The cells were treated with maximum non-cytotoxic doses of Tet (for CNE1 cells at 1.5 μmol/L and for CNE2 cells at 1.8 μmol/L),irradiation at 4 Gy,or combination of irradiation and maximum non-cytotoxic doses of Tet.The cell cycle distribution was analyzed by flow cytometry.The protein levels of γ-H2AX,cleaved caspase-3,p-CDC25C,CDK1,p-CDK1,cyclin B1,ERK and p-ERK were determined by Western blot.RESULTS:The expression of γ-H2AX was increased in CNE1 cells and CNE2 cells after combined treatment with irradiation and maximum non-cytotoxic doses of Tet.The percentages of CNE1 cells and CNE2 cells at G2/M phase in irradiation group were (18.09 ±0.42)％ and (18.48 ± 1.32)％,respectively,which were decreased to (15.88 ± 1.04) ％ and (13.80 ± 0.82) ％ in combined treatment group,respectively (P ＜ 0.05).Combined treatment enhanced the increase in the protein level of cleaved caspase-3 caused by irradiation.The protein levels of pCDC25C and p-CDK1 were increased in a dose-dependent manner by Tet treatment (P ＜ 0.05),while the expression of CDK1 showed no difference among different doses of Tet treatments.The protein levels of p-CDC25C,p-CDK1 and CDK1 showed no difference after the treatment with maximum non-cytotoxic doses of Tet.The combined treatment with irradiation and the maximum non-cytotoxic doses of Tet decreased the protein levels of p-CDC25C and p-CDK1 (P ＜0.05),increased the expression of cyclin B1,and had no influence on the expression of CDK1 （ P ＜0.05).The combined treatment resulted in an increase in the protein level of p-ERK1 (P ＜ 0.05).CONCLUSION:The maximum non-cytotoxic doses of Tet enhance the DNA damage and apoptosis in CNE1 cells and CNE2 cells caused by irradiation,and the mechanism might be associated with ending of G2/M arrest via activation of ERK/CDC25C/CDK1/cyclin B1 pathways.

China ; Genetic Testing ; Glucosephosphate Dehydrogenase ; genetics ; Glucosephosphate Dehydrogenase Deficiency ; genetics ; prevention & control ; Humans ; Mutation