A quantitative analysis method of multi-components with a single marker (QAMS) for simultaneous determination of six marker compounds (one from phenolic acids and five from phthalides) in Chuanxiong Rhizoma was established by applying HPLC and using butylidenephthalide as the internal reference substance. And also the feasibility and accuracy of the established method for quality evaluation and application of Chuanxiong Rhizoma were investigated and validated. The analysis was performed with the mobile phase consisting of acetonitrile - 0.2% aqueous formic acid. The flow rate was 1.0 mL . min-1 and the column temperature was maintained at 30 °C. The detection wavelengths were set at 252 nm (for ferulic acid, Z-ligustilide, and butylidenephthalide) and 266 nm (for senkyunolide I, senkyunolide A, and coniferyl ferulate), separately, and 20 µL was injected for analysis with gradient elution. The results showed that there were no significant differences observed between the HPLC-QAMS method and the external standard method (RSD <5%). The relative correction factors were credible (RSD < 5%) in changed chromatographic conditions. The established HPLC-QAMS method can be accurately used for simultaneously evaluating and controlling the quality of Chuanxiong Rhizoma with multi-components.
4-Butyrolactone ; analogs & derivatives ; Acetonitriles ; Benzofurans ; Chromatography, High Pressure Liquid ; Coumaric Acids ; Drugs, Chinese Herbal ; analysis ; standards ; Hydroxybenzoates ; Quality Control ; Rhizome ; chemistry

OBJECTIVE Despite the status of cisplatin (DDP) as a classical chemotherapeutic agent in the treatment of cancer, the development of multidrug resistance often leads to a failure of DDP therapy.Traditional Chinese medicine(TCM)as adjuvant chemotherapy of cancer drugs in China has been widely used in cancer treatment.ZuoJin WAN (ZJW),a TCM formula,was proved reversing drug resistance in gastric cancer,but its exact mechanism was still unclear. METHODS CCK-8 assay was used to detect the cell viability. The levels of proteins and mRNA were evaluated using Western blot and q-PCR. Mitochondrial membrane potential was measured by fl ow cytometry. Depolymerisa-tion of F-actin and translocation of G-actin(gamma-actin)from the cytoplasm to the mitochondria was detected using an immuno fl uorescence assay. RESULTS phosphorylated coflin-1 (p-coflin-1) was overexpressed in the DDP-resistant human gastric cancer cell lines SGC7901/DDP and BGC823/DDP, relative to the respective parent cell lines(SGC7901 and BGC823),and DDP induced the dephosphory-lation of p-coflin-1 in both parent lines but not in the DDP-resistant lines. However, ZJW could induce the dephosphorylation of pcoflin-1 and promote coflin-1 translocation from the cytoplasm into the mito-chondria in both SGC7901/DDP and BGC823/DDP cells. This mitochondrial translocation of coflin-1 was found to induce the conversion of flamentous actin to globular-actin, activate mitochondrial dam-age and calcium overloading, and induce the mitochondrial apoptosis pathway. These effects of ZJW on DDP-resistant human gastric cancer cell lines could be reversed via transfection with coflin-1-specifc siRNA,or treatment with a PP1 and PP2A inhibitor.CONCLUSION ZJW can be used as an inhibitor of chemoresistance in gastric cancer, which may partly be due to dephosphorylation of p-coflin-1 via the activation of PP1 and PP2A.

Objective To observe the effect of iodine excess(HI),polyinosinic-polycytidylic acid[Poly(I:C),Poly]and thyroglobulin(TG)on the thyroid of mice by the expression of Toll-like receptor 3(TLR3)to reveal the functional role of TLR3 in autoimmune thyroiditis.Methods Forty-two non-obese diabetic mice,body weight (20±3)g,were divided into six groups:control group,HI group,Poly group,TG group,HI+TG group,HI+Poly group. Fed with deionized water and injected intraperitoneally with physiological saline 0.1 ml each day for a week, the mice in control group were injected with physiological saline every other day at the same dose for 1 week before they were sacrificed; HI group drank 0.05% NaI water and were injected intraperitoneally with physiological saline same as control group; Poly group drank deionized water and were injected intraperitoneally with poly 0.1 ml (1 g/L)each day of the week, then the mice were injected with Poly every other day at the same dose for 1 week before they were sacrificed; TG group drank deionized water and were injected intraperitoneally with physiological saline same as control group, immunized with 0.1 mg TG by subcutaneously injecting and the immunization was enhanced after they were fed half dose for 4 and 8 weeks separately. In HI + Poly group, the treatment was the same as HI group and Poly group; HI + TG group: the treatment was the same as HI group and TG group. Eight weeks later, mice were sacrificed and thyroids were taken to make frozen sections, Hematoxylin-Eosin (HE) staining was employed to observe the morphological change of the thyroids. The expression of TLR3 of thyroids was observed under fluorescence microscope after Immumofluorescence using TLR3 antibody and TR3-positive cells were analyzed in the thyroid density. Results HE staining showed thyroids of Poly group had no inflammation under microscope.There were different degrees of inflammatory cell infiltration in HI group and TG group. The inflammatory cell infiltration and the damage of follicular thyroid of HI + TG group and HI + Poly group were serious, and the degrees of inflammation were higher over "++". Thyroid follicular epithelial cell with TLR3 expression could be seen in Poly group and HI group, meanwhile, there were TLR3 strong positive inflammatory cells in HI group under fluorescent microscope. Using stereological analysis of TLR3-positive cell density in the thyroid, the difference between groups was statistically significant(F=7.870, P<0.01 ). TLR3-positive cell density in the thyroid of HI + Poly group was higher[ (9.287 ± 0.522)mm2] than control group[ (0.062 ± 0.025)mm2, P < 0.01] significantly, meanwhile, the density in HI + Poly group was higher than HI group [ (2.574 ± 0.257 )mm2] and Poly group[ (1.361 ± 0.148 )mm2, all P < 0.01]. The density in HI + TG group[ (4.843±0.405)mm2] was higher than HI group and TG group[(1.601 ±0.268)mm2, all P < 0.01 )]. Conclusions Excessive iodine and thyroglobulin can induce thyroiditis, and stimulate the expression of TLR3 in the thyroid follicular epithelial, Poly aggravated thyroiditis induced by iodine excess in NOD mice; TLR3 positive inflammatory cells also appeared in inflammatory region, suggesting that TLR3 is involved in the pathogenesis of autoimmune thyroiditis

Anti-Bacterial Agents ; therapeutic use ; Child ; Child, Preschool ; Drug Resistance, Bacterial ; Humans ; Mycoplasma pneumoniae ; drug effects ; pathogenicity ; Pneumonia ; drug therapy ; microbiology

Objective To study the pharmacokinetic and pharmacody-namic characteristics of febuxostat tablets after single and mutiple oral dose in hyperuricemia patients with chronic kidney disease .Methods A total of 18 hyperuricemia patients with chronic kidney disease were ran-domly divided into three groups , each group containing 6 patients as fol-lows: those in group A were administrated with a single oral dose of febuxostat tablet 40 mg, those in group B were administrated with a single oral dose of febuxostat tablet 80 mg, and followed by multiple oral doses of febuxostat 80 mg, qd for 28 days and those in group C were treated with a single oral dose of febuxostat tablet 120 mg.The serum uric acid was determined by clinical chemistry testing , and the plasma drug levels were determined by HPLS -MS/MS method.Results The main febuxostat pharmacokinetic parameters of a single oral administra-tion of 40, 80 and 120 mg division were as follows: t1/2 were (3.98 ±1.45), ( 4.63 ±1.37 ) and ( 4.27 ±1.33 ) h; tmax were (0.97 ±0.39), ( 1.22 ±0.46 ), ( 1.35 ±0.37 ) h; Cmax were (1626.33 ± 342.97 ) , ( 2669.27 ± 384.91 ) , ( 3904.83 ± 835.98 ) ng ? mL-1; AUC0-24 h were (4324.32 ±976.87), (8415.4 ±1965.9), (12979.6 ±3788.5) ng? h? mL -1.Drug accumulation was not detec-ted following multiple oral doses .After 4 weeks treatment (80 mg, qd) , febuxostat lowered the serum uric acid levels of hyperuricemia patients with chronic kidney disease significantly , and the target serum uric acid level (6.0 mg? dL -1 ) was achieved in 5 patients.Conclusion Febuxostat displayed a linear pharmacokinetic profile in single oral doses of 40 to 120 mg.Continuous oral therapic dose of febuxostat tablets can significantly reduce the uric acid levels in hyperu -ricemia patients with chronic kidney disease .

OBJECTIVETo explore the impact of environmental factors, daily lifestyle, psycho-social factors and the interactions between environmental factors and chemokines genes on systemic lupus erythematosus (SLE).

METHODSCase-control study was carried out and environmental factors for SLE were analyzed by univariate and multivariate unconditional logistic regression. Interactions between environmental factors and chemokines polymorphism contributing to systemic lupus erythematosus were also analyzed by logistic regression model.

RESULTSThere were nineteen factors associated with SLE when univariate unconditional logistic regression was used. However, when multivariate unconditional logistic regression was used, only five factors showed having impacts on the disease, in which drinking well water (OR=0.099) was protective factor for SLE, and multiple drug allergy (OR=8.174), over-exposure to sunshine (OR=18.339), taking antibiotics (OR=9.630) and oral contraceptives were risk factors for SLE. When unconditional logistic regression model was used, results showed that there was interaction between eating irritable food and -2518MCP-1G/G genotype (OR=4.387). No interaction between environmental factors was found that contributing to SLE in this study.

CONCLUSIONMany environmental factors were related to SLE, and there was an interaction between -2518MCP-1G/G genotype and eating irritable food.

Adult ; Case-Control Studies ; Chemokines ; biosynthesis ; genetics ; China ; epidemiology ; Female ; Genetic Predisposition to Disease ; genetics ; Humans ; Logistic Models ; Lupus Erythematosus, Systemic ; epidemiology ; genetics ; Male ; Polymorphism, Genetic ; Risk Factors ; Surveys and Questionnaires

OBJECTIVETo determine the correlation of the CYP1A1 (rs4646422) gene polymorphisms with male infertility in the Chinese Han population.

METHODSUsing the Mass ARRAY iPLEX GOLD technique, we conducted a case-control study on theCYPlA1 (rs4646422) gene polymorphisms in 636 infertile males aged 21-49 years (case group) and 442 normal healthy men aged 23-47 years (control group) of the Chinese Han population. We analyzed the genotypes and allele frequencies in the two groups ofsubjects with the SPSS 20.0 software.

RESULTSCompared with the wild homozygous genotype GG, the heterozygous genotype AG (OR = 1.06, 95% CI 0.81-1.38) and homozygous genotype AA (OR = 1.11, 95% CI 0.56-2.21) showed no correlation with male infertility, nor did the mutant allele A (OR = 1.06, 95% CI 0.85-1.32) in comparison with the wild allele G.

CONCLUSIONThe CYP1A1 (rs4646422) gene polymorphisms might not be correlated with male infertility in the Chinese Han population.

Adult ; Alleles ; Case-Control Studies ; China ; Cytochrome P-450 CYP1A1 ; genetics ; Gene Frequency ; Genotype ; Homozygote ; Humans ; Infertility, Male ; genetics ; Male ; Middle Aged ; Polymorphism, Genetic ; Young Adult

Esophagustype cervical spondylosis is easily misdiagnosed or missed diagnosed for its lower incidence. The pathological basis manily concerned with compression of osteophyte for esophageal and tracheal wall and local inflammatory reaction. The diagnosis of disease is generally not difficult. Especially dynamic esophagogram could not only show degree of cervical degeneration and osteophyte shape, but also could clearly observe location and degree of osteophytosis indenting esophageal and tracheal. Recently, the treatment of esophagustype cervical spondylosis has been the focus of attention. It is generally believed that conservative treatment could alleviate disease but could not cure it. More and more scholars tend to treat disease by surgical resection of osteophyte to relieve compression for esophagus, and clinical symptoms could obviously improved. However, long-term follow up is lack, and further follow-up effect is lack of powerful support, anddifferent treatment methods and surgical procedures are lack of comparative research.

OBJECTIVETo initially evaluate the application of artificial vertebra of n-HA/PA66 in anterior reconstruction of lower cervical spine fracture and dislocation.

METHODSIn this study, 84 patients with lower cervical spine fracture and dislocation received anterior cervical discectomy, spinal canal decompression or subtotal corpectomy, spinal canal decompression and reconstruction by n-HA/PA66 composite artificial vertebral body combined with plate instrumentation. Neurological function was followed up by improvement rate of Frankel and situations of the supporting body was observed by X ray and 3D-CT in 3, 12, 24 months postoperatively. The intervertebral height, physical arc (reflected by Cobb angle) and the locations and fusion rate of the supporting body were assessed in order to evaluate the stability of the cervical spine and alignment improvements.

RESULTSAll the patients underwent operation successfully and were followed up for 6 to 24 months with an average of 12 months. The preoperative symptoms were improved to varying degrees. Imaging studies showed that in all cases graft fusion were achieved, and cervical alignments, intervertebral height, cervical spine stability and the locations of the artificial vertebral body were well maintained. No displacement and subsidence of the artificial vertebral body occurred. Postoperative immediate intervertebral height (2.4 ± 0.2) cm, preoperative intervertebral height (1.9 ± 0.1) cm, comparisons of the two groups was statistically significant (q = 2.48, P < 0.001). The immediate, 3 month, 1 year, 2 year period follow-up group intervertebral height was not statistically significant (P > 0.05). Preoperative Cobb angle was 9.8° ± 1.2°, postoperative immediate Cobb angle was 16.6° ± 1.2°, comparisons of the two groups was statistically significant (q = 14.25, P < 0.001). The immediate, 3 month, 1 year, 2 year period follow-up group Cobb angle was not statistically significant (P > 0.05).

CONCLUSIONSn-HA/PA66 artificial vertebral body can provide early cervical spine support and stability and effectively maintain the biological alignment and cervical intervertebral height. It has high rate of graft fusion and is convenient to observe by X-ray. Therefore, n-HA/PA66 can be taken as an ideal graft for anterior lower cervical spine fracture and dislocation operation, but further follow-up study is still required to evaluate the long-term effects.

Adolescent ; Adult ; Aged ; Bone Substitutes ; Cervical Vertebrae ; injuries ; surgery ; Decompression, Surgical ; Female ; Follow-Up Studies ; Fracture Fixation, Internal ; Humans ; Hydroxyapatites ; Joint Dislocations ; complications ; surgery ; Male ; Middle Aged ; Nanostructures ; Nylons ; Spinal Fractures ; complications ; surgery ; Spinal Fusion ; instrumentation ; Young Adult

The sialyl Lewis X (SLex) antigen encoded by the FUT7 gene is the ligand of endotheliam-selectin (E-selectin).The combination of SLex antigen and E-selectin represents an important way for malignant tumor metastasis.In the present study,the effect of the SLeX-binding DNA aptamer on the adhesion and metastasis of hepatocellular carcinoma HepG2 cells in vitro was investigated.Reverse transcription-polymerase chain reaction (RT-PCR) and immunofluorescence staining were conducted to detect the expression of FUT7 at both transcriptional and translational levels.The SLex expression in HepG2 cells treated with different concentrations of SLeX-binding DNA aptamer was detected by flow cytometry.Besides,the adhesion,migration,and invasion of HepG2 cells were measured by cell adhesion assay,and the Transwell migration and invasion assay.The results showed that the FUT7 expression was up-regulated at both mRNA and protein levels in HepG2 cells.SLeX-binding DNA aptamer could significantly decrease the expression of SLex in HepG2 cells.The cell adhesion assay revealed that the SLeX-binding DNA aptamer could effectively inhibit the interactions between E-selectin and SLex in the HepG2 cells.Additionally,SLeX-binding DNA aptamers at 20 nmol/L were found to have the similar effect to the monoclonal antibody CSLEX-1.The Transwell migration and invasion assay revealed that the number of penetrating cells on the down-side of Transwell membrane was significantly less in cells treated with 5,10,20 nmol/L SLeX-binding DNA aptamer than those in the negative control group (P＜0.01).Our study demonstrated that the SLeX-binding DNA aptamer could significantly inhibit the in vitro adhesion,migration,and invasion of HepG2 cells,suggesting that the SLeX-binding DNA aptamer may be used as a potential molecular targeted drug against metastatic hepatocellular carcinoma.