A UPLC method has been developed in the current investigation for simultaneous determination of nine chemical markers of Bletilla striata, 4-hydroxymethylphenyl beta-D-glucoside, blestroside, dactylorhin A, militarine, dihydrophenanthrene 5, gymnoside V, dihydrophenanthrene 1, benzylphenanthrene 3 and gymnosides IX. Separation was performed at 45 degrees C on an ACQUITY UPLC BEH C18 column (2.1 mm x 150 mm, 1.7 microm) with a gradient solvent system of acetonitrile-water as the mobile phase. The flow rate was 0.3 mL x min(-1), the detection wavelength was 280 nm. The results showed that the nine chemical markers could be well resolved and that in the selected linear range, all calibration curves of the nine chemical markers showed good linearity (r > or = 0.999 3). The recoveries (n = 6) were in the range of 98.15% - 102.2% and RSDs were between 2.1% - 3.6%. The data suggested that the developed UPLC-UV method had good reproducibility, robustness, and accuracy, which was suitable for the quality control of Bletilla striata. Applications of the method showed that the nine chemical markers had higher contents in the wild B. striata than in the cultivated ones.
Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; isolation & purification ; Magnoliopsida ; chemistry ; Rhizome ; chemistry

Objective To investigate the effects of saturated fatty acids and unsaturated fatty acids on proliferation and autophagy of human lung cancer cells. Methods The lung cancer cells A549 were treated with stearic acid (saturated fatty acid) and doconexent (DHA, unsaturated fatty acid), respectively, in concentrations of 0, 30, 60, 120 and 240μmol/L. MTT test and cell clone formation assay were performed to detect the proliferation of A549 cells. The morphology of A549 autophagy was observed by confocal laser scanning microscopy after A549 cells were treated with stearic acid or DHA for 24 hours. Western blotting assay was used to detect the expression of autophagy-related protein after A549 cells were treated with stearic acid or DHA for 12, 24 and 36 hours, respectively. Results 30-240μmol/L stearic acid or DHA both inhibited the proliferation of A549 cells (P<0.05). Both stearic acid and DHA induced autophagy of A549 cells, meanwhile, down-regulated Phospho-mTOR (ser2481) and up-regulated LC3Ⅱ/LC3Ⅰ of A549 cells (P<0.05). Conclusions Both saturated fatty acid and unsaturated fatty acid can inhibit the proliferation and induce autophagy of lung cancer cells. The mechanisms of autophagy may be related to Phospho-mTOR (ser2481) signaling pathway.

Objective To investigate the supply of iodized salt in non-excessive iodine counties and iodine-free salt in excessive iodine counties at household level in Jiangsu Province so as to provide a basis for the prevention and control of iodine deficiency disorders(IDD).Methods According to the National Iodine Deficiency Disorders Monitoring Program(Trial),county(city,district) was taken as a elementary sampling unit in Jiangsu Province.Townships(towns) and administrative villages were selected by systematic sampling and random sampling in every county and households were chosen by random sampling to collect their edible salt samples.The salt iodine content in non-and excessive iodine regions was detected by direct titration method and semi-quantitative method,respectively.Results All 30 840 salt samples were collected from 106 non-excessive iodine counties,and qualified iodized salt was 30 303 copies,iodine-free salt 199 copies.Weighted by the population of counties,the rate of iodine-free salt was 0.71％,the coverage rate of iodized salt accounted for 99.29％,out of which,98.93％ was qualified and the consuming rate of qualified iodized salt was 98.23％.All 1296 salt samples were collected in 5 counties with excessive water iodine content and the coverage rate of iodine-free salt was 98.99％ (1283/1295).Conclusions The national targets for preliminary elimination of IDD have been achieved in regions of nonexcessive and excessive iodine of Jiangsu Province.But it still should be strengthen the management work of iodinefree salt in excessive iodine counties and iodine saft in non-excessive iodine counties.

OBJECTIVETo explore the relationship between genetic polymorphism of P53, P21, CCND1 and susceptibility of chromosomal damage induced by vinyl chloride monomer (VCM).

METHODS183 workers occupationally exposed to VCM were involved in our study. Cytokinesis-block micronucleus (CB-MN) assay was used to detect chromosome damage in peripheral lymphocyte. PCR-RFLP technique was applied to detect polymorphisms in P53 gene (exon4, intron3 and intron6), P21 gene (exon2 and exon3) and CCND1 (exon4).

RESULTSThe risk of chromosomal damage for VCM-exposed workers with more than 30 yr was 1.2202 (95% CI: 1.0580 approximately 1.4072, P = 0.0062) compared with the younger workers, and the risk of female workers was 1.1491 (95% CI: 0.9841 approximately 1.3416, P = 0.0772) compared with male workers. The MN frequency in subjects with P53 intron6 mutant homozygous and heterozygous was higher than their wild-type homozygous counterparts (OR = 1.3032, 95% CI: 1.1285 approximately 1.6405, P = 0.0285). P53 exon4, intron3 and intron6 haplotype pairs of BBB/AAA and BAB/AAA were associated with the increased frequencies of micronucleus.

CONCLUSIONAmong VCM-exposed workers, more than 30ys, female, carrying P53 intron6 mutated allele and BBB/AAA and BAB/AAA haplotype pairs have higher risk of chromosomal damage.

Cell Cycle Checkpoints ; Humans ; Micronucleus Tests ; Occupational Exposure ; Polymorphism, Genetic ; Vinyl Chloride

OBJECTIVETo study the method of induced eruption on multiple adjacent impacted teeth in anterior maxillary bone.

METHODSTwenty-two multiple adjacent impacted teeth of 9 cases were chosen. The position of the impacted teeth and the relationship to each other were assessed on X-ray images, oral examination and plaster model. The impacted teeth were extracted or induced erupted with the closed-eruption technique and fixed orthodontic appliances.

RESULTSOf 22 impacted teeth, 19 impacted teeth were moved into arches by induced eruption with the closed-eruption technique and fixed orthodontic appliances. There were not root adsorption or conglutination for 19 impacted teeth. Three impacted teeth with deformed root were extracted. The average time for treatment was 19 months.

CONCLUSIONWith correct diagnosis, reasonable design, the complicated impacted teeth could be moved into alignment with good esthetic and functional effect.

Cuspid ; Humans ; Maxilla ; Orthodontic Appliances ; Tooth Eruption ; Tooth Root ; Tooth, Impacted

OBJECTIVETo establish the specific 16S-23S rRNA gene spacer regions pattern in different bacteria using polymerase chain reaction (PCR), restriction fragment length polymorphism (RFLP), DNA cloning and sequences analysis.

METHODSA pair of primers were selected from highly conserved sequences adjacent to the 16S-23S rRNA spacer region. Bacterial DNA of sixty-one strains of standard bacteria and corresponding clinical isolates representative of 20 genera and 27 species was amplified by PCR, and further studied by RFLP, DNA cloning and sequences analysis. Meanwhile, all specimens were examined by bacterial culturing and PCR-RFLP analysis.

RESULTSThe 27 different standard strains showed one, two, three or more than three bands. The sensitivity of PCR reached 2.5 colony-forming unit (CFU), and there was no cross reaction to the human, fungal or viral genomic DNAs. Fifteen species could be distinguished immediately by PCR, while another 10 species were further identified by Hinf I or Alu I digestion. Klebsiella pneumoniae (Kp) and Enterococcus durans (Ed) could not be differentiated from each other by Alu I or Hinf I digestion. The spacer sequences of the Kp and Ed were 908 bp and 909 bp, respectively, and they differed only at the site of the 779th nucleotide. The former was G, and the latter was A. The 760 - 790 bp sequence of Kp was as follows: CGACTGCACCGCCTCCTAC / GGCCGCGTATTC. The 760 - 790 bp sequence of Ed was as follows: CGACTGCAC CGCCTCCTAC / AGCCGCGTATTC. Only one enzyme XmaIII, could discriminate the two. The cleaving site of XmaIII is C downward arrow GGCCG. Kp DNA was cleaved into 778 bp and 130 bp fragments, while E. durans was not. Of 42 specimens with suspected septicemia, 15 were positive (35.7%) on blood culture, and 27 on PCR (64.29%). The positive rate of PCR was significantly higher than that of blood culture (P < 0.01). Of the six CSF specimens, one was positive for Staphylococcus epidermidis (Se) on culture as well as by PCR, while two specimens which were negative on cultures were positive by PCR and were diagnosed as Se according to its DNA pattern. One specimen was culture-positive for Cryptococcus neoformans (Cn) but was negative by PCR. The other two specimens were negative by both PCR and culture. Fifteen blood samples from healthy children were negative by both blood culture and PCR.

CONCLUSIONSThe method of detecting bacterial 16S-23S rRNA spacer regions using PCR-RFLP techniques was specific, sensitive, rapid and accurate in detecting pathogens in clinical bacterial infections.

Bacterial Infections ; diagnosis ; microbiology ; DNA, Bacterial ; chemistry ; genetics ; DNA, Ribosomal Spacer ; genetics ; Humans ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; RNA, Ribosomal, 16S ; genetics ; RNA, Ribosomal, 23S ; genetics ; Sequence Analysis, DNA

OBJECTIVETo observe and compare the curative effect of pingyangmycin and sodium morrhuae respectively in the treatment of oral and maxillofacial venous malformation.

METHODSForty cases of oral and maxillofacial venous malformation were induced in this study, 20 of which were given local injection of low dose of pingyangmycin, and 20 given local injection of 5% sodium morrheate. The therapeutic effects were observed.

RESULTSThe effective cases of pingyangmycin was 19, and the effective cases of sodium morrhuate was 12. The side effect was found in 10 cases of, and in 15 cases sodium morrhuate group, the former was mainly pingymangmycin group systemic, the later was mainly local. The difference was significant between the two groups (P < 0.05).

CONCLUSIONSPingyangmycin is more effective in effective rate and side effect treatment of venous hemangioma than sodium morrhuate.

Adolescent ; Adult ; Bleomycin ; administration & dosage ; analogs & derivatives ; therapeutic use ; Child ; Child, Preschool ; Female ; Follow-Up Studies ; Humans ; Infant ; Injections, Intralesional ; Male ; Middle Aged ; Sclerosing Solutions ; therapeutic use ; Sodium Morrhuate ; administration & dosage ; therapeutic use ; Treatment Outcome ; Vascular Malformations ; therapy ; Young Adult

OBJECTIVETo describe the prevalence of anemia and yearly trends (1993-2003) among women who came to the hospitals or maternal and child health units for premarital examinations in 6 counties of Jiangsu and Zhejiang provinces.

METHODSData were obtained from the records of the premarital examinations in perinatal health care surveillance system that had been established since 1992 in these areas. We reviewed hemoglobin levels of those women who were enrolled in the perinatal health care surveillance system from 1993 to 2003. Anemia was defined according to the WHO (2001) criterion. We calculated the prevalence of anemia and analyzed the yearly trends based on the data of hemoglobin concentration.

RESULTSIn the period of 1993-2003, there were 82 995 anemia cases identified among 193,434 women with an overall anemia rate as 42.9%. The rates of anemia were high (65.5%) in 1993 but low (25.8%) in 2003. 99.7% of the anemic women whose hemoglobin concentration were between 80-119.9 g/L. Time trend analysis indicated a significant decline on anemia rate while monthly analysis showed that the prevalence rates were high (48.2%) in September and low in March (39.5%). The results also showed that the prevalence rates of anemia were relatively higher in farmers and workers in rural enterprises, and lower in Han ethnicity than minorities. The higher prevalence rates of anemia were presented among the women with less education, lower body mass index, or at older age.

CONCLUSIONFor those premarital women in 6 counties of Jiangsu and Zhejiang provinces, the overall anemic rate presented a significant downward trend between 1993 and 2003 while the prevalence of anemia remained high, especially for the women with less education, lower body mass index or older ages.

Adolescent ; Adult ; Anemia ; epidemiology ; China ; epidemiology ; Female ; Humans ; Middle Aged ; Premarital Examinations ; Young Adult

Objective To investigate the intervention effect of Xuebijing Injection on the differentiation of bone marrow hematopoietic stem cells (HSC) in septic mice. Methods Fifty-four male C57BL/6N mice were randomly divided into three groups: normal control group, model group and Xuebijing group, each group with 18 mice. The mouse models of sepsis were duplicated by intra-peritoneal injection of 10 mg/kg E. coli lipopolysaccharide (LPS) method. Starting from the day of modeling, Xuebijing Injection 20 mL/kg was intravenously injected into the tail vein in Xuebijing group, once a day for consecutive 4 days; the normal control and model groups were intravenously injected with normal saline at the same dose and site for 4 days. The bone marrow cells of the femur and tibia of the mice were isolated after 4 days of various treatments in the three groups, and the proportions of bone marrow HSC Lin-Sca-1+c-Kit+ (LSK) and hematopoietic progenitor cells Lin-Sca-1-c-Kit+ (LS-K) of each group were detected by flow cytometry. Results Finally, 14 mice were included in the normal control group, 17 in the model group, and 12 in the Xuebijing group. With the prolongation of time, the body weight of the normal control group gradually increased, the body masses of the model group and the Xuebijing group were decreased first and then increased, reaching a peak at 96 hours after the model was established, but they were still significantly lower than the body mass of normal control group (g: 19.81±0.27, 19.58±0.39 vs. 22.23±0.30, both P < 0.05). Compared with the normal control group, the proportions of LSK, LS-K, long-term HSC (LT-HSC), and megakaryocyte-erythroid progenitor cells (MEP) were all significantly increased in the model group [LSK: (16.62±1.28)% vs. (12.89±0.83)%, LS-K: (44.77±1.77)% vs. (30.34±0.90)%, LT-HSC: (6.88±0.48)% vs. (1.83±0.24)%, MEP: (13.89±1.26)% vs. (9.38±0.66)%, all P < 0.05], the proportion of multipotential progenitor cells (MPP) was significantly decreased [(2.41±0.34)% vs. (5.99±0.59)%, P < 0.05]. Compared with the model group, the LSK and myeloid progenitor (CMP) of the Xuebijing group were significantly reduced [LSK: (12.25±0.69)% vs. (16.62±1.28)%, CMP :(0.31±0.05)% vs. (0.55±0.13)%, both P < 0.05], and LS-K, LT-HSC, MEP showed a decreasing trend [LS-K: (42.75±2.48)% vs. (44.77±1.77)%, LT-HSC:(5.98±0.70)% vs. (6.88±0.48)%, MEP: (10.94±1.36)% vs. (13.89±1.26) %], but the differences were not statistically significant (all P > 0.05). There were no significant differences in the proportions of short-term HSC (ST-HSC) and granulocyte-macrophage progenitor cells (GMP) among the three septic groups (all P > 0.05). Conclusion Xuebijing Injection can improve the differentiation function of bone marrow cells in septic mice, which may be possibly related to the inhibition of pathological proliferation of bone marrow hematopoietic stem cells and progenitor cells in septic mice by Xuebijing Injection.

OBJECTIVE: To examine the use of PCR utilizing 16S-23S rRNA gene spacer regions in the identification of bacteria. METHODS Primers used in PCR were designed by using the target sequences from the genes encoding 16S-23S rRNA spacer regions. PCR was used for the detection of different standard and clinical bacterial strains. RESULTS Characteristic DNA maps were present after using the PCR to identify 27 standard strains from 27 species. The maps could be directly used for classification of the tested bacterial strains or further differentiated by RFLP. The sensitivity of the PCR may be as high as 2.5 CFU/ml. No non-specific amplification products were observed when using DNA from human PBMC funguses or viruses as templates. Thirty-two strains of bacteria isolated from clinical strains showed DNA maps similar to the DNA maps amplified from standard strains. CONCLUSION The PCR detection of bacteria using 16S-23S rRNA gene spacer regions is sensitive, rapid, specific and accurate for identification of bacteria and provides a new rapid method for determining the clinical diagnosis and the etiology of sepsis.