This article reviewed the appli ca tion of molecular biology technology, such as genomics, DNA labeling technology, and DNA microarray. The aim is to provide the reference for the modernization o f Chinese material medica.

Aim To establish insulin resistant HepG2 cell model induced by high concentration insulin in vitro culture and evaluate the effects of pioglitazone on insulin sensitivity and glucose metabolism in the cell model.Methods HepG2 cells were incubated with 5?10~(-7)mol?L~(-1) insulin for 16 h and insulin-stimulated()~3H-D glucose incorporation was determined.Then,insulin-stimulated glucose incorporation rate which was used to estimate insulin sensitivity of HepG2 cell,and glucose consumption,the amountsof glucose disappeared from the culture medium of HepG2 cells within 24 hours was determined.The insulin resistance cells were incubated with pioglitazone 1?10~(-5) mol?L~(-1).Results It was demonstrated that the incorporation rate of()~3H-D-glucose in insulin-resistant HepG2 cells was decreased significantly than that in the control cells.Insulin-resistant HepG2 cells were free from stimulation for 48 h by insulin,but the incorporation rate of()~3H-D-glucose was lower than that in control cells.Incubation of insulin resistance cells with pioglitazone significantly increased glucose uptake and glucose consumption by the cells compared with that of the control cells(P

Aim To investigate the effects of ecdysterone on insulin sensitivity and glucose metabolism in the insulin resistant HepG2 cell model induced by high concentration of insulin. Methods The insulin-stimulated glucose incorporation rate was determined with ~3H-D-glucose uptake test which was used to estimate insulin sensitivity of HepG2 cell, and glucose consumption, the amounts of glucose disappeared from the culture medium of HepG2 cells within 24 h were determined. Results The incubation of insulin resistance cells with ecdysterone 1?10~ -6 ～10~ -4 mol?L~ -1 could significantly increase the glucose uptake and glucose consumption of the cells compared with that of control cells(P0.05). Conclusion Ecdysterone could improve the insulin sensitivity in the cell model and can attenuate the aggression of insulin resistance. The effect to improve the insulin sensitivity with ecdysterone is similar to that with pioglitazone.

Aim To investigate whether ecdysterone is able to exert glucose-lowering effect on hepatocytes or stimulate the secretion of insulin.Methods For glucose consumption studies,the amounts of glucose disappeared from the culture medium of HepG2 cells within 24 h were determined.?TC3 cells were also used to monitor insulin secretion.Results The glucose concentrations decreased significantly by ecdysterone 1?10~(-6)～10~(-4) mol?L~(-1),while glucose consumption increased by 44%～77% with ecdysterone.Glucose consumption declined as its concentrations increased.Insulin had no effect on glucose-lowering of ecdysterone.?TC3 cells were not stimulated by ecdysterone.Conclusion Ecdysterone is able to exert glucose-lowering effect on hepatocytes which is insulin independent,but has no effect on insulin secretion.

Objective To explore the relationship between the expression of P53 protein and the radiosensitivity in patients with advanced maxillary squamous cell carcinoma.Methods An immunohistochomical method wag used to detect the expression of P53 protein in patients with advanced maxillary squamous cell carcinoma.The follow up time was 2 years.The local recurrence of the patients having been treated with radical surgery and affiliated radiotheraphy were analyzed.Results The overexpression of P53 protein in 26 cages was 65.4% (17/26).In the P53 overexpression group,the local recurrence after systiem therapy wag occurred in one case within 6 months,6 cages between 7～12 months.3 cages between 13～18 months and 2 cases between 19～24 months.In the P53 low expression group,there were no recurrence within 6 and 12 months and one case ocurred within 18 months.5 cages between 19～24 months.The difference of recurrence within 18 months after system therapy between the expression of P53 Wag statistically significant(P＜0.05),but it Wag not significant for those within 24 months(P＞0.05).Conclusions The expression of P53 protein Wag correlated with the radiosensitivity in patient with advaneed maxillary squamous cell carciaoma,especially for the resid.rod cells in mitosis phase.The affiliated radiotherapy after radical surgery Wag limited effect.

Purpose　The aim is to investigate the hemostatic and protective effect of fibrin sealant(FS) on liver trauma in rats.Methods　On the surface of cracked wound(type Ⅰ) and resected wound(type Ⅱ)，spraying of FS or thrombin or direct suturation, was used receptively. Natural hemostasis was used as control.During postoperation the hemostatic time and the state of wound healing on 1st, 4th, 7th,14th, 50th day were observed.Results　Compared with the natural hemostasis,the hemostatical time of the FS group was shorter 86.0%(P＜0.01)in liver trauma type Ⅰ and 79.0%(P＜0.01)in liver trauma type Ⅱ.Compared with the thrombin group,the hemostatical time of the FS group was shorten 45.0%(P＜0.01)in liver trauma type Ⅰ and 84.0%(P＜0.05) in liver trauma type Ⅱ.The wound surface of FS group was healed faster than that of other groups.Conclusion　FS was an effective hemostatic and healing-promoter on liver trauma of rats.

Objective To isolate specific humanized anti-D-dimer scFv(single chain Fv) antibody from scFv phage libraries. Methods Isolate anti-D-dimer positive clones from Tomlinson I + J phage libraries by three rounds of panuing, then sequence monoclonal genes by bideoxy-mediated chain termination and express soluble scFv antibody; Pick out anti-D-dimer antibodies with high specificity and affinity by ELISA.Results After three rounds of selection from human scFv phage libraries Tomlinson I and J, 38 monclonal specific anti-D-dimer scFv fragments were selected. By polyclonal and monoclonal phage ELISA and gene sequencing, 20 different full-length monoclonal scFv phages were identified, the result of soluble scFv ELISA showed that 20 full-length monoclonal scFv were expressed smoothly. According to the result of soluble scFv ELISA, in 5 scFv antibodies with high value of A450 selected, 3 scFv antibody fragments showed high specific and affinity. Conclusion Antibody phage display was an effective, rapid method to isolate anti-D-dimer antibodies with high specificity and affinity.

Objective To investigate an association between Toll-like receptor(TLR)4 Asp299Gly or CD14 C-260T polymorphisms and colorectal cancer in Chinese patients.Methods By a method of polymerase chain reaction-based restriction fragment length polymorphism(PCR-RFLP), TLR4 Asp299Gly and CD14 C-260T polymorphisms in unrelated 110 patients with colorectal cancer and 160 healthy controls in Chinese Han race population were genotyped.Results Significant differences of CD14 genotypes were found between healthy controls and the patients with colorectal cancer.The fre- quency of the CC genotype in healthy controls was extremely lower than that in the group of colorectal cancer(15.6% vs.31.8%,P=0.0027,OR=0.3968,95%CI:0.2209-0.7129)while the frequency of the CT genotype in healthy controls was significantly higher than that in the group of colorectal cancer (48.1% vs.30.9%,P=0.0056,OR=2.074,95%CI:1.246-3.452).No TLR4 Asp299Gly mutant genotype was detected in any patients and healthy controls in Chinese Han population.Conclusion The polymorphism of CD14 C-260T but not TLR4 Asp299Gly is associated with colorectal cancer in Chinese patients.

BACKGROUND:It is a great potential study that calcium sulfate product loaded with antibiotics is developed, but this product is not systematicaly studied and its biocompatibility and security need to be further studied.
OBJECTIVE:To evaluate the biocompatibility and safety of vancomycin- or gentamicin-loaded calcium sulfate bone.
METHODS: (1) Hemolysis test: vancomycin-loaded, gentamicin-loaded calcium sulfate extracts, double distiled water and saline were added into rabbit anticoagulant blood samples. (2) Micronucleus test: vancomycin-loaded and gentamicin-loaded calcium sulfate extracts, cyclophosphamide and normal saline solution were intraperitonealy injected to mice, respectively. (3) Acute toxicity test: vancomycin-loaded and gentamicin-loaded calcium sulfate extracts, and normal saline solution were intraperitonealy injected to mice, respectively. (4) Pyrogen test: the mice were injected vancomycin-loaded and gentamicin-loaded calcium sulfate extractsvia the ear vein. (5) Intradermal stimulation test: vancomycin-loaded and gentamicin-loaded calcium sulfate extracts were respectively injected into the unilateral spine of rabbits, respectively. (6) Intramuscular implantation test: vancomycin-loaded and gentamicin-loaded calcium sulfate extracts were respectively injected to the dorsal muscle of rabbits. (7) Intraosseous implantation test: vancomycin-loaded and gentamicin-loaded calcium sulfate were implanted into the necrotic femoral bone of rabbits.
RESULTS AND CONCLUSION:Both vancomycin-loaded and gentamicin-loaded calcium sulfate products, which have no hemolytic reaction, genetic toxicity, acute toxicity, pyrogen reaction and skin irritation, are considered to have good biocompatibility and safety.

Objective To classify the taxonomic status of O.cheni in relation to O.turkestanicum var. tuberculata from the mainland of China by comparing their nucleotide sequences of nuclear ribosomal partial large subunit gene (LSU). Methods The genomic DNA of adult worms were extracted by the GNT-K method. The target gene was amplified by PCR using specific primers. The PCR products were purified before ligation into the plasmid PCR-blunt (Invitrogen). Recombinant plasmids were amplified in E.coli , extracted and purified using routine methods and then sequenced using M13 primers (F/R) on a Licor long-read auto-sequencer. Sequences of O. turkestanicum was retrieved from GenBank and aligned with our data in BioEdit. Results The nucleotide sequences of LSU between O.turkestanicum var. tuberculata and O.cheni was 100% identical, and 99.99% identical between O.turkestanicum var. tuberculata and O.turkestanicum . Conclusion This study demonstrated high similarity in LSU nucleotide sequences, and the results do not support O.cheni as an independent species. O.cheni may be a synonym of O.turkestanicum var. tuberculata , and O.turkestanicum var. tuberculata is probably also a synonym of O.turkestanicum .