OBJECTIVETo study the influence of different drying methods on the quality of Schisandrae Chinensis Fructus and thus provide useful reference for its proper drying methods.

METHODSchisandrae Chinensis Fructus was processed by eight drying methods including vacuum freeze drying, natural drying in the shade, drying in the sun, oven drying and vacuum drying under different temperature. The contents of the functional ingredients includes chisandrin, gomisin D, gomisin J, schisandrol B, angeloylgomisin H, angeloylgomisin Q, gomisin G, schisantherin A, deoxyschisandrin, schisandrin B, schisandrin C, 5-HMF, total aids and total sugars. The main components change after drying were analyzed by HPLC, ultraviolet spectrophotometry and potentiometric titration. Principal component analysis (PCA) was carried out by SPSS software to evaluate the quality of different processed products from Schisandrae Chinensis Fructus.

RESULTAll these results are in accordance with the requirements of Chinese Pharmacopoeia published in 2010, the contents of schisandrin and total eleven lignans were the highest using vacuum drying, and 5-HMF were the lower, oven drying made little difference but with lower schisandrin and higher 5-HMF as the heat increased.

CONCLUSIONDifferent drying methods have significant influence on the quality of Schisandrae Chinensis Fructus. Oven drying under 5°C should be adopted to substitute drying in the sun according to the China Pharmacopoeia published in 2010 for Schisandrae Chinensis Fructus by comprehensive analysis of the cost, content and practicality.

Desiccation ; methods ; Drugs, Chinese Herbal ; chemistry ; Quality Control ; Schisandra ; chemistry ; Temperature

The ginsenoside-Ro is sensitive to be hydrolyzed in an alkaline medium. This paper investigated the hydrolysis kinetics of ginsenoside-Ro under different pH and temperature values. The results showed ginsenoside-Ro in alkaline solution followed pseudo-first-order reaction. Hydrolysis kinetics of ginsenoside-Ro has not been reported previously. The hydrolysis rate was independent of initial concentration. On the basis of UFLC-MS/MS, NMR, as well as chemical evidence,the structure of hydrolyzate was assigned as 3-O- [beta-D-glucuronopyranosyl- (1 --> 2) -beta-D-glucopyranosyl] -oleanolic acid.
Drugs, Chinese Herbal ; chemistry ; Ginsenosides ; chemistry ; Hydrogen-Ion Concentration ; Hydrolysis ; Kinetics ; Magnetic Resonance Spectroscopy ; Mass Spectrometry ; Molecular Structure ; Spectrometry, Mass, Electrospray Ionization

In gene manipulation, different selectable markers with various linkers are necessary. In order to get selectable markers directly, we constructed from pBlueScript SK(-) a series of particular plasmids, pSKsymKm, pSKsymBle, pSKsymEry, pSKsymHyg and pSKsymGm, each contains Kanamycin, Bleomycin, Erythromycin, Hygromycin or Gentamycin resistance cassette. By restriction enzyme digestion and gel extraction, any of five antibiotic resistance genes with specific ends can be conveniently obtained.

Objective To explore the effect of different processing and drying methods of corn and hot pepper on fluorine content in coal-burning type of the endemic fluorosis areas, and to screen food processing and drying methods which meet the quality requirements of grain drying and able to effectively reduce the total fluoride intake of local population. Methods Farmers of endemic fluorosis area in Bijie, Guizhou province were divided into 3 groups: sun-baked drying group, stove drying group with air-tight cover and stove drying group with no cover, 10 households in each group. Corn and fresh hot pepper and samples dried for 2 weeks, or 1, 3, 6-month were collected, and water and fluoride content were detected, and the total daily fluoride intake were calculated in accordance with the "Determination of Water in Food" (GB/T 5009.3-2003) and "Determination of Fluorine in Foods"(GB/T 5009.18-2003). Results Fluoride content in fresh corn and dried for 2 weeks, or 1, 3, 6-month [of sunbaked drying group: (1.40 ± 0.16), (1.56 ± 0.14), (2.15 ± 0.47), (2.70 ± 0.64), (4.06 ± 1.75)mg/kg, stove drying group with air-tight cover: (1.41 ± 0.16), (2.39 ± 0.56), (4.60 ± 0.97), (8.46 ± 5.55), (11.36 ± 3.60)mg/kg,stove drying group with no cover: (1.40 ± 0.13), (4.69 ± 3.97), (4.47 ± 2.77), (9.65 ± 6.47), (26.12 ± 14.52)mg/kg] and pepper[sun-baked drying group: (5.41 ± 1.61), (16.60 ± 7.62), (32.60 ± 7.88), (50.26 ± 17.60),(240.20 ± 272.49)mg/kg, stove drying group with air-tight cover: (754 ± 2.95), (3238 ± 11.50), (119.18 ± 156.45),(224.00 ± 196.58), (495.70 ± 417.29)mg/kg, stove drying group with no cover: (4.82 ± 1.25), (44.30 ± 13.48),(122.89 ± 66.43), (334.23 ± 166.05), (531.01 ± 397.40)mg/kg] increased with elongation of drying time, and the group difference was significant(F = 44.77, 128.71, 126.87, 41.61, 53.63, 170.63, all P ＜ 0.05), with the largest rate of increase in stove drying group with no cover, and the lowest in sun-baked drying group;fluoride was significantly lower (t = 7.93,63.07,5.36,11.98,55.76,7.45, all P ＜ 0.05) after sample washing;total fluoride intake per person per day was 2.57 mg in local adult when ate washed and sun-baked corn, peppers, the total fluoride intake were 5.92, 8.14 mg when ate the food processed by other two drying methods and washed corn, peppers,respectively. Conclusions In the coal-burning type of fluorosis endemic area, should take appropriate health education measures, and instruct local residents to use sun bake their edible corn and pepper for human consumption, and cultivate a habit of washing corn and pepper before cooking, which can reduce the population total fluoride intake, and control endemic fluorosis.

Objective To evaluate the effects of implementing the health education in the Central Fund Program to control endemic fluorosis in Guizhou.Methods The samples were randomly surveyed to evaluate knowledge awareness in students and households as well as the habit formation after implementing the integrated program which mainly consisted of installing the improved stoves,supported by the Central Funds and health education in 5 counties.Results After health education,the rates of knowledge awareness in the students and the households were 94.80%(15 562/16 415)and 88.23%(4482/5080),respectively,and increased significantly compared with those before intervention[44.20%(26 364/59 645),22.81%(3082/13 510)],the difference being significant(χ2=13 324.05,6546.24,P＜0.01).The rates of drying corn and chili with the coal fire were 5.61% (57/1016)and 5.41%(55/1016),respectively,and decreased significantly compared with those before intervention [77.41%(1076/1390),78.92%(1097/1390)],the difference being significant(χ2=1214.49,1270.92,P＜0.01).The rates of washing corn and chili were 99.51%(1011/1016)and 94.59%(961/1016),respectively,and increased significantly compared with those before intervention[84.60%(1176/1390),76.55%(1064/1390)],the difference being significant(χ2=154.80,143.32,P＜0.01).The rates of using the uncovered and unventilated iron stoves and table stoves were 4.71%(38/807)and 8.37%(60/717),respectively,and decreased significantly compared with those before intervention[29.99%(14 483/48 299),98.33%(95 070/96 685)],the difierence being significant(χ2=243.51,25 282.99,P＜0.01).Conclusions Implementing the health education is the basis for the integrated measures for controlling the endemic fluorosis in the endemic regions.The consciousness and activity of the target people have been enhanced greatly.The good behaviors in the target people are forming,the expected goal is reached.

Objective To explore the influence of stoves and mode of coal-burning on the prevalence of endemic fluorosis in Guizhou Province.Methods Three previous severe endemic villages and two mild villages(as control)were investigated by ways of random sampling in the present and retrospect epidemiological study on stove usage,coal burning mode and the severity of disease.Results Before the 1990's Of 20th century all the households in the previous severe endemic villages used the open stove without ventilated pipe(chimney)and dehydrated the corn over the coal-fire of the open stoves in house,which resulted in severe pollution of the corn and indoor air by fluoride.After the 1990's,each household,although still dried the corn with the coal fire,used the table stoves and iron stoves increasingly year by year,which resulted in the decrease of the emitted fluoride from the coal fire significantly,reduced the fluoride pollution of the indoor air and the main food,thus the incidence of endemic fluorosis consequently decreased. On the other hand, most of the households in the previous severe endemic villages such as Lianhe and Yuanzi still used the open, unventilated stove without chimney, very few installed chimney leading to outside of the room,the rates being 51.3%(41/80)and 41.7%(35/84),respectively.It was concordant to the high incidence of endemic fluorosis.Compared with 1979,the detective rates of dental fluorosis in the crowd aged above 8 years old were increased significantly in Hehua Village in 2006,90.51%(248/274)and 96.08%(392/408),respectively(χ2=7.85,P＜0.01).Conclusions The usage of stoves and mode of coal-burning significantly influence the incidence of the endemic fluorosis caused by domestic coal-burningin Guizhou Province,and inappropriated use of stove and the mode of coal burning are the key factors in cause the prevalence of the severe fluorosis in endemic regions in Guizhou.

OBJECTIVETo clone the gene coding for the peptidoglycan recognition protein (PGRP) domain (PGRPd) of mouse long PGRP (mPGRP-L) and express the protein in E. coli.

METHODSThe cDNA fragment encoding PGRPd of mPGRP-L was obtained by RT-PCR from the total RNA of Balb/C mouse liver cells and cloned into pUCm-T vector. The recombinant plasmid were identified by PCR, restriction endonucleases and sequence analysis. The PGRPd gene fragment was amplified by PCR from the recombinant plasmid, inserted into pQE-30 vector and transformed into E. coli strain M15, and the expressed PGRPd protein was purified.

RESULTSA cDNA fragment of about 500 bp was amplified by RT-PCR and the recombinant plasmid, pmPGRPd, was constructed by linking the fragment to pUCm-T vector. The results of restriction mapping of the recombinant vector were consistent with those of computer analyses. Sequence analysis showed that the cloned gene fragment (518 bp) had identical sequence with the gene encoding PGRPd of mPGRP-L gene in GenBank. The recombinant expression vector pQE-PGRPd was constructed and expressed in E. coli M15. SDS-PAGE showed that the expressed product existed mainly in the lysate supernatant as a soluble protein with relative molecular mass of 29 kD.

CONCLUSIONThe PGRPd cDNA of mPGRP-L has been successfully cloned and expressed in E. coli, which provides the basis for further study of PGRP molecule.

Animals ; Base Sequence ; Carrier Proteins ; genetics ; Cloning, Molecular ; DNA, Complementary ; genetics ; Escherichia coli ; genetics ; metabolism ; Female ; Liver ; chemistry ; Mice ; Mice, Inbred BALB C ; Molecular Sequence Data ; Plasmids ; genetics ; Prokaryotic Cells ; metabolism ; Protein Structure, Tertiary ; genetics ; Recombinant Proteins ; biosynthesis ; genetics

This study was aimed to investigate the distribution and implication of tap1 (transporter associated with antigen processing) and tap2 loci allelic and genotypic frequencies. The distribution of tap1 and tap2 loci allelic and genotypic frequencies in 339 random samples of healthy Chinese Hans was analyzed by TaqMan PCR. Several genetic information about power of discrimination, cumulative DP, polymorphism information content, expected heterozygosity and observed heterozygosity were calculated. The results indicated that 5 tap1 alleles (tap1*0101, 020101, 020102, 0301 and 0401) and 4 tap2 alleles (tap2*0101, 0102, 0103 and 0201) were detected in all samples. 8 tap1 genotypes were found which account for 53.3% of the theoretic genotype and 6 tap2 genotypes were found which account for 60% of the theoretic genotype. The genotyping results of tap1 and tap2 both conform to the Hardy-Weinberg expectations (p > 0.05). Tap1*0101 (79.79%) and tap2*0101 (82.74%) are the most common alleles in Chinese Hans. It is concluded that tap1*0101 and tap2*0101 are most common alleles in Chinese Hans, tap1 and tap2 loci carry some power of individual discrimination and polymorphism information content. These two locl can be used for the research in the fields of human genetics, linkage analysis of genetic disease genes, paternity test and individual identification and so on.
ATP-Binding Cassette Sub-Family B Member 2 ; ATP-Binding Cassette Transporters ; genetics ; ATP-Binding Cassette, Sub-Family B, Member 3 ; Alleles ; Asian Continental Ancestry Group ; genetics ; Gene Frequency ; Genotype ; Haplotypes ; Humans

OBJECTIVETo investigate the plasma levels of ascorbic acid and vitamin E in patients with liver cirrhosis and to explore their significance.

METHODSThe plasma levels of ascorbic acid,vitamin E and lipoperoxides in patients with liver cirrhosis were measured, and the results were compared with those of sex-and age-matched healthy subjects.

RESULTThe plasma levels of ascorbic acid, vitamin E and lipoperoxides in the patients group were (42.94 +/-6.99)micromol/L, (17.99 +/-3.51)micromol/L and (14.09 +/-1.28)micromol/L, respectively, while those in the control group were (53.30 +/-9.45)micromol/L (t=9.50, P=0.000), (24.59 +/-7.22)micromol/L (t=7.94, P=0.000) and (12.11 +/-1.20)micromol/L (t=17.21, P=0.000), respectively.

CONCLUSIONThe levels of ascorbic acid and vitamin E in patients with liver cirrhosis decrease significantly,which may indicates the disturbance of balance between oxidation and antioxidation.

Adult ; Aged ; Aged, 80 and over ; Ascorbic Acid ; blood ; Female ; Humans ; Lipid Peroxides ; blood ; Liver Cirrhosis ; blood ; metabolism ; Male ; Middle Aged ; Vitamin E ; blood

OBJECTIVETo explore the pattern of hair cell injury and expression of P53 apoptosis protein in intensive impulse noise injured cochlear hair cells in guinea pigs.

METHODSTwelve adult guinea pigs were exposed to a series of 40 pairs of impulse noise (2 second intervals) at the intensity of 168 dB (SPL). Animals were terminated at 3, 6 and 12 hours after noise exposure, respectively. Cochlear surface preparations were performed with a double staining of FITC-conjugated phalloidin and propidium iodide for the observations of the stereocilia and the nucleus. P53 immunochemical staining was also performed 12 hours post-noise exposure to observe if there was expression of p53 protein in injured hair cells. Results Three hours after noise exposure, the outer hair cells at the end of basal turn and beginning of second turn were destroyed first with a character of nuclear condensation. Six hours post-noise exposure, many hair cells in the center of damage region had nuclear fragmentations, and the damaging area expanded towards to basal turn and apical turn. Twelve hours after noise exposure, the nucleus in most outer hair cells and inner hair cells at the region of damage center were missing. The nuclear condensation and fragmentation were appeared in hair cells in both sides of the center region of degeneration. P53 immunoreactive products were also found in damaged hair cells, not only in the central damage area, but also in the basal turn and the third turn.

CONCLUSIONSIntensive impulse noise resulted in apoptosis of cochlear hair cells that initiated between the end of basal turn and the beginning of second turn. Hair cell degeneration spread to basal and third turn along the basilar membrane. P53 may play an important role in impulse noise induced-hair cell apoptosis.

Animals ; Apoptosis ; Cochlea ; metabolism ; pathology ; Guinea Pigs ; Hair Cells, Auditory ; metabolism ; pathology ; Hearing Loss, Noise-Induced ; metabolism ; pathology ; Noise ; adverse effects ; Tumor Suppressor Protein p53 ; metabolism