Objective In order to study the value of drug treatment for leukemia, quercetin was delivered to the cultured NB4 cell line. MethodsUsing RT- PCR and western blot, we studied the expression of mutant p53 gene and protein after inducing 24, 48, 72, 96 and 120h by the quercetin with different concentrations (30, 60, 90μmol/L). Results We verified the results that quercetin with concentration of 30 ～90μmol/L could successfully inhibit the expression of mutant p53 protein, but the mutant p53 gene did not have. ConclusionThat data suggested that quercetin would be an effective method for the therapy of leukemia.

Apolipoprotein A5(ApoA5)level and other indices were determined in patients with type 2 diabetes mellitus and healthy individuals.Compared to control group,ApoA5 level in the diabetic group was lower (P

ObjectiveTo prepare high specific monoclonal antibodies(mAbs) against BP26 of Brucella(B.)melitensis.Methods A recombinant plasmid pET-28a-BP26 was constructed and transformed into competent Escherichia coli BL21 (DE3),and then the bacteria were induced by 1 mmol/L isopropylthio-β-D-galactoside (IPTG).After induction,the recombinant BP26 protein (rBP26) was purified by sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PGAE) and nickel ion affinity chromatography(Ni-NTA).Mice were inoculated with rBP26 antigens for three times at 2-week intervals.The first subcutaneous injection contained 100 μg rBP26 with 0.1 ml complete Freund adjuvant.The second subcutaneous injection was 50 μg rBP26 with 0.1 ml incomplete Freund adjuvant.The antibody titers to rBP26 were determined 2 weeks after each reimmunization.Three days before cell fusion,the mice with the highest titer were intraperitoneally injected with 50 μg rBP26 in 0.1 ml PBS.Pre- and post-immunization sera were collected and used as negative or positive controls for screening mAbs.Mice with the highest titer were sacrificed and spleen cells were isolated.The spleen cells of rBP26 immunized mice were fused with SP2/0 myeloma cells in a ratio of 5 ∶ 1 by polyethylene glycol(PEG) 1450.Antibody-producing hybridomas were primarily screened by an indirect enzyme-linked immunosorbnent assay(ELISA) with rBP26.Reactive hybridomas were subcloned for 3 times,then the strains of hybridoma cells secreting antibodies against BP26 were obtained.Supernatant of cloned hybridoma cultures was collected for mAb analyses.These mAbs were named by the hybridoma clone number and tested their reactivity to membrane proteins extracted(NMP) from B.melitensis vaccine strain(M5-90) by Western blotting and Dot-ELISA.mAbs isotyping and kappa(κ) or lambda(λ) light chain was identified by Mouse Monoclonal Antibody Isotyping Kit.Results A total of two mAbs reactive to rBP26 of B.melitensis were selected from antibody screening hybridomas by indirect-ELISA.The two mAbs were named 3C3 and 5A5,and identified as IgG1 (κ) and IgG2(κ),respectively.They could react with NMP from M5-90.Conclusions Results of identification show that two mAbs against rBP26 can be produced.The two mAbs can recognize natural BP26 protein,giving the experimental materials for further research on identification of its epitopes.

AIM:To explore the involvement of oxidative stress and apoptosis in the pathogenesis of diabetic cataract induced by Streptozotocin ( STZ) and the interventions of puerarin in order to supply references for clinical treatment.
METHODS:Male SD rats were divided into four groups randomly, control group, diabetic group, apocynin group and puerarin group. The diabetic group were replicated by single injection of STZ (65mg/kg, ip). The expression of p22, p47, p67, Bax/Bcl2, Caspase 3 and P53 proteins were detected by Western Blotting.
RESULTS:The diabetic rats were replicated successfully and the expression of Bcl2 was downregulated while the expression of p22, p47, p67, Bax, Caspase 3 and P53 were upregulated in diabetic group with a significant statistical differences when compared with control group (P<0. 05). Apocynin and prerarin can reverse the abnormal expression of the aforementioned proteins dramatically (P<0. 05).
CONCLUSION: NADPH oxidase mediated oxidative stress and P53, Bax/Bcl2 mediated apoptosis are involved in the pathogenesis of diabetic cataract and puerarin can alleviate cataract greatly by inhibiting the aforementioned signal pathway.

Spinal biomechanics, especially the range of spine motion,has close connection with spinal surgery. The change of the range of motion (ROM) is an important indicator of diseases and injuries of spine, and the essential evaluating standards of effect of surgeries and therapies to spine. The analysis of ROM can be dated to the time of the invention of X-ray and even that before it. With the development of science and technology as well as the optimization of various types of calculation methods, diverse measuring methods have emerged, from imaging methods to non-imaging methods, from two-dimensional to three-dimensional, from measuring directly on the X-ray films to calculating automatically by computer. Analysis of ROM has made great progress, but there are some older methods cannot meet the needs of the times and disappear, some classical methods such as X-ray still have vitality. Combining different methods, three dimensions and more vivo spine research are the trend of analysis of ROM. And more and more researchers began to focus on vivo spine research. In this paper, the advantages and disadvantages of the methods utilized recently are presented through viewing recent literatures, providing reference and help for the movement analysis of spine.
Animals ; Humans ; Imaging, Three-Dimensional ; instrumentation ; methods ; trends ; Radiography ; Spine ; diagnostic imaging

A method for qualitative analysis of constituents in Panax notoginseng by UPLC-LTQ-Orbitrap mass spectrometry was established. Based on the high-resolution mass information, MS/MS fragmentation behaviors and chemical components from literatures, 43 compounds were identified or tentatively characterized. New type saponin aglycone, combined with malonyl-substituted and acetyl-substituted saponins were discovered and plausibly identified in this study. This work could be helpful for the quality control and further phytochemical studies of Panax notoginseng, and provided a good example for the analysis of chemical constituents in traditional Chinese medicine.

This study aimed at evaluating the quality of the precise powder decoction pieces (PPDP) of L.japonicae Flos (LJF) compared with the traditional commercial slices with chemical fingerprint methods and DNA molecular identification technology.Different specifications of PPDP were prepared,their dry extract contents were in contrast with that of commercial slices.The three batches of commercial slices were collected,and the content uniformity,fingerprint and similarity evaluation before and after the mixing and pulverization were studied by HPLC-DAD and DNA sequence alignment.As a result,the paste rate of PPDP was slightly higher than that of the traditional commercial slices.The dissolution of chlorogenic acid of PPDP was higher than that of the traditional commercial slices.RSD of inter-assay dissolutions of chlorogenic acid of commercial slices was 11.93％,which was reduced to 8.29％ after mixing and preparing into PPDP.The fingerprint showed that the slimilarity of the fringerprint of the mixed and powdered LJF was elevated with 7 common peaks.All the common peaks were increased at different levels.In conclusion,compared with traditional commercial slices of LJF,PPDP apparently improved the dissolution rate and the quality uniformity,indicating that the boiled powder of CRP obviously presented vantages in clinic.

This study aimed at evaluating the quality of precise powder decoction pieces (PPDP) of E.Folium (EF) compared with the traditional commercial slices by chemical fingerprint methods and DNA molecular identification technology.Different specifications of PPDP were prepared,and their dry extract contents were in contrast with that of commercial slices.The slices of EF were identified using ITS2 and psbA-trnH sequences.Three batches of commercial slices were collected,and the content uniformity,fingerprint and similarity evaluation before and after the mixing and pulverization were detected by HPLC-DAD and DNA sequence alignment.It was found that the paste rate of PPDP was slightly higher than that of the traditional commercial slices.The dissolution of chlorogenic acid of PPDP was higher than that of the traditional commercial slices.RSD of inter-assay dissolutions of chlorogenic acid of commercial slices was 15.56％,which was reduced to 6.82％ after mixing and preparing into PPDP.The fingerprints showed that the similarity of the PPDP of EF was elevated with the inceases of 10 marketed common peaks.The PPDP of EF was accurately identified by ITS2 and psbA-trnH sequences.In conclusion,compared with traditional commercial slices of EF,the PPDP apparently improved the dissolution rate and the quality uniformity,demonstrated that the boiled powder of CRP achieved obvious clinic advantages.

In this study,DNA molecular identification technology and chemical fingerprint method were adopted to evaluate the quality system of precise powder decoction pieces (PPDP) of S.suberectus dunn (SSD).ITS2 sequence was taken as DNA barcode to indentify SSD.Different specifications of PPDP were prepared,their dry extract contents were quantified in contrast with that of original slices.Three batches of SSD original slices were gleaned and the content uniformity,fingerprint and similarity evaluation before and after the mixing and pulverization were valued by HPLC-DAD.As a result,ITS2 successfully and accurately identified the SSD in this study.The extract rate of PPDP was 15.5％,1.11 times as much as the original slices.RSD of inter-assay dissolution of cepicatechin from the original slices was 11.0％,which was reduced to 1.0％ after mixing and preparing into PPDP.The relative peak area of the 14 common peaks identified by fingerpringts were larger,while the RSD values significantly decreased.It was concluded that the PPDP of SSD improved the extraction efficiency and uniformity of the original slices,featuring quite prospective in more reasonable and scientific clinical use.

Objective To investigate the clinical efficacy of superficial needling therapy for early ankylosing spondylitis and its effect on tumor necrosis factor-α (TNF-α), interleukin-1 (IL-1) and interleukin-6 (IL-6).Methods Eighty patients with early ankylosing spondylitis meeting the inclusion criteria were randomly allocated to treatment and control groups using random number card method with the ratio of 1:1, 40 cases each. The treatment group received superficial needling therapy alone and the control group, conventional treatment with celecoxib capsules, methotrexate tablets and sulfasalazine enteric-coated tablets. Treatment was given for a total of 24 weeks. Bath AS Disease Activity Index (BASDAI), Bath AS Functional Index (BASFI), lumbago, TCM syndrome score, TNF-α, IL-1 and IL-6 were observed in the two groups before and after treatment.Results The BASDAI, BASFI, lumbago, TCM syndrome score, TNF-α, IL-1 and IL-6 improved in the two groups after treatment compared with those before the treatment (P<0.05). The BASDAI, BASFI, lumbago, TCM syndrome score, TNF-α, IL-1 and IL-6 in the treatment group superior to those in the control group after the treatment (P<0.05).Conclusions Superficial needling therapy can relieve the clinical symptoms in patients with early ankylosing spondylitis and improve spinal joint function with good safety. Its mechanism may be related to reducing inflammatory cytokines TNF-α, IL-1 and IL-6.