Objective: To study the scutellaria and coptis in different proportions, the main active ingredient of baicalin, berberine dissolution rate changes and to explore the effect of traditional drug pair of scutellaria and coptis on the dissolution rate of active ingredient. Methods: Based on the clinic common used prescriptions of scutellaria with coptis, ratio of 1:0,1:1,1:2,1:3,2:1,2:3,3:1,3:2,0:1 was selected, after the water boiling reflux extraction, refining separation, various products were obtained for the test, under the optimized conditions of RP-HPLC analysis to compare chromatographic fingerprints and to examine the main component baicalin, berberine in aspects of relative peak area ratio of changes and compatibility relations. Results: Compatibility of Scutellaria baicalensis and Coptis at different proportions can influence the dissolution rate of baicalin, Berberine Hydrochloride. There was a non-linear relationship of dissolution rate of baicalin and Scutellaria baicalensis. The best ratio of scutellaria ratio was 3:1, the best compatibility ratio of coptis was 1:3. Conclusion: Different compatibility proportions of scutellaria and coptis will result in different dissolution rate of baicalin and berberine.

AIM: To explore a simple, reliable method for tissue processing and section staining by extracting DNA from the manually microdissectioned specimen, and to identify whether the extracted DNA is useful in the following study at molecule level. METHODS: The experiment was performed at the pathological laboratory of Guangdong Medical College from July 2004 to July 2007. The paraffin imbedding tissue sections of cervical cancer were thoroughly deparaffinized after mounted on slides for a long period of time. The nucleus was slightly stained with hematoxylin and microdissectioned under inverted microscope. The microdissectioned samples were put into EP tubes filled with digestion buffer to split the cells and then the DNA was extracted. During the whole course, PE tubes did not change, and the complicated phenol/chloroform extraction did not perform. The DNA extraction was rapid and simple. RESULTS: The DNA was measured by the spectrophotometer with concentrations from 0.14 to 5.25 g/L and absorbance values of A260/A280 were 1.6-1.8. All samples were amplified with PCR to produce expected length specific target fragment (231 bp). CONCLUSION: Rapid DNA extraction after manual tissue microdissection can produce adequate amount of DNA and maintain good quality of DNA template for PCR. The DNA meets the need of the following molecular experiments.

Objective To obtain two types of optical isomers from 1-(Benzamidomethyl)-1,2,3,4-tetrahydro-isoquinoline (BTIQ). Methods ( ±) BTIQ as raw materials , optically pure camphor sulfonic acid as resolution agent was used and repeated resolution in acetone , And the resolution product was detected by specific optical rotation .The product of BTIQ was hydrolyzed , and the ratio of ATIQ was compared with that of the literature .ResuIts After two repeated chemical resolutions , the specific rotations of both enantiomers are no longer changed .It was showed that the products of higher optical purity.The specific rotation of both isomers are -35.65°(CH2Cl2, C=0.5)with the yield of 27.52%,and +35.17°(CH2Cl2, C =0.5) with the yield of 31.55% respectively.The specific rotation value of chiral 1,2,3,4-tetrahydroisoquino line ( ATIQ) which was the hydrolysis product of BTIQ consistent with values reported in the literature .ConcIusion The (-) BTIQ and ( +) BTIQ enantiomers were successfully obtained by the method of resolution , and the yield and optical purity of the obtained products are higher , laying the foundation for the further development of high efficiency , low toxicity of chiral schistosomicide ( praziquantel ) and other containing tetrahydroisoquinoline structure of chiral drugs .

Objective To detect the changes in the expression of discoidin domain receptor 2(DDR2)and matrix metalloproteinase (MMP)-13 in different stages of cartilage and synovium damage of osteoarthritis rats.The relation between DDR2 and the degree of cartilage damage was explored.Methods Modified papain knee joint injection approach was adopted to establish animal model of OA.The expression and distribution of protein of DDR2 and MMP-13 were checked in articular cartilage and synovium at different stages of OA.Results The expressions of DDR2 in articular cartilage and synovium of experimental groups were different from those of the normal group (P＜0.01).They were higher in cartilage than those in the corresponding synovium.The expressions of MMP-13 demonstrated the same characteristics with those of DDR2,r=0.93(P＜0.01).Conclusions The important role of DDR2-MMP-13 in cartilage damage has been proven in the pathogenic process of OA.The upregulated expressions of DDR2 in articular cartilage and synovium have a detrimental effect on cartilage degeneration.

Objective To explore the gene sequencing and prenatal diagnosis of Glanzmann thrombasthenia (GT). Methods The blood samples were drawn from one case of phenotype GT pediatric patient, patient’s parents, and one normal control. The amniotic lfuid and cord blood from the fetus of patient’s mother were collected. When the fetus was born 2 days, the blood was drawn. The coagulation routine test and platelet aggregation test were performed. The expression of platelet membrane glycoprotein (GP) IIb and GPIIIa were tested by lfow cytometry. Microsatellite technology is used to determine whether fetal cord blood is contaminated with maternal cells. The expressed region and the junctional zone between exon and introns of GPIIb and GPIIIa were ampliifed by PCR technology from blood sample of patient, patient’s parents, and fetus’s cord and 2 days after birth. The PCR products were then subjected to DNA sequencing. Results Adenosine diphosphate (ADP) cannot induce the platelet aggregation in the patient. The max rate of the platelet aggregation in the fetus’s cord blood was half of the normal. However, the max aggregation rate induced by ADP in the blood sample of parents and fetus 2 days after birth were equal to normal. The mean lfuorescence intensity (MnX) of platelet membrane GPIIb and GPIIIa in the patient were 10%and nearly zero of the normal control, respectively, while those in the parents, the fetus’s cord blood and 2 days after birth were more than 90%and 30%to 50%of the normal control. The cast-off cells in amniotic lfuid and the DNA in cord blood analysis by microsatellite technology conifrmed that the amniotic lfuid and cord blood not contaminated by maternal cells. Gene analysis showed the heterozygosis mutation in exon6 A3829→C and exon9 G42186→A of the patient’s GPIIIa led to the amino acid heterozygosis mutation in GPIIIaHis281→Tyr and Cys400→Pro. These two mutations came from the father and the mother separately. However, there was only one heterozygosis mutation in exon9 G42186→A in the cast-off cells in amniotic lfuid, the fetus’s cord and blood 2 days after birth. Conclusion This GT patient have double heterozygosis mutation. The fetus has heterozygosis mutation conifrmed after birth.

Objective To test and evaluate the geometric accuracy of delineation of organs at risk ( OARs) in head and neck cancer using an atlas?based autosegmentation ( ABAS) software. Methods The atlases for the ABAS software was generated using images from 40 patients with head and neck cancer undergoing intensity?modulated radiotherapy. The software was tested in 40 new patients. Automatic delineation of OARs was carried out on computed tomography images by single?( one to one ) and multi?template ( ten to one) approaches. In order to evaluate the feasibility of the automatic delineation in clinical application, differences in volume (ΔV%), position (Δx,Δy, andΔz), conformability (sensitivity ( Se ), specificity ( Sp ) , and dice similarity coefficient ( DSC) ) , and delineation time were assessed between the automatic and manual delineation. The comparison between the two automatic delineation approaches was made by paried t test. Results For all OARs, the multi?template automatic delineation achieved a significantly smaller mean ΔV% value and a significantly larger mean DSC value than the single?template automatic delineation (-0.02%± 0?29% vs. -0.16%± 0?41%, P<0?05;0.74± 0?16 vs. 0.68± 0?20, P<0?05);the position differences between two automatic delineation approaches were less than 0?4 cm in all three directions except for the temporal lobe, lower jaw, and spinal cord;in the receiver operating characteristic curve defined by Se versus 1-Sp , the data points were all within the first quadrant except for the optic nerve and chiasm;automatic delineation saved 42%?72% of time compared with manual delineation. Conclusions The ABAS software achieves satisfactory results of automatic delineation for most of OARs in patients with head and neck cancer. The multi?template automatic delineation, particularly, has better outcomes than the single?template one. In addition, it greatly shortens the time the clinicians spend on delineation of OARs.

Objective:To study the efficacy and safety of natural calcined bone repair material(NCBM)in repairing bone defect af-ter tooth extraction.Methods:A randemized,double-blinded,parallel,positive control(Bio-Oss)and multi-center clinical trial was employed.Imaging examination was used as the main efficacy evaluation index,surgical wound healing,rejection reaction,bone me-tabolic changes,bone infection signs were the subordinate efficacy evaluation indexes,the incidence of adverse reactions was observed for safety evaluation.Results:280 cases were included,269 cases completed the trial.In NCBMand Bio-Oss group the effective rate of imaging examination was 93.08% and 93.70%(P >0.05)respectively.The wound healing time of the 2 groups was less than 7 days,no rejection reaction,bone metabolic change and bone infection sign were observed.The incidence of adverse events in NCBM and the Bio-Oss group was 0.72% and 2.14%(P >0.05)respectively.Conclusion:The efficacy and safety between natural cal-cined bone repair material is not inferior to Bio-Oss in repairing bone defect after tooth extraction.

Objective To study the regulatory effect of triptolide(TP)on the angiogenesis of coll-agen-induced arthritis(CIA) rats.The effect of TP on arthritis is also explored.Methods After the model of CIA was established,the articular volume was measured and the synovium was examined with regular HE stainand the inflammation and pathological changes were evaluated.In addition,the vascular endothelial growth factor (VEGF),basic fibroblast growth factor (bFGF)and endostatin protein expressions in synovium and serum were tested.The micro-vessel density (MVD) of synovium was also measured by caulating CD34 level.Results The expressions of VEGF,bFGF and MVD in CIA rats'synovium and serum were evidently higher than the control group(X2=65.3,31.6,q=9.2,P＜O.01,respectively),while the expression of endostatin showed no statistical difference with controls (X2=0.8,P＞0.05).After treated with TP,the expressions of VEGF,bFGFand MVD decreased markedly(X2=19.7,6.0,q=6.5,P＜O.01,respectively),but the pmtein expression of endostatin significantly increased (X2=3.9,P＜O.05).However,only the expression of endostatin increased significantly after treated with MTX (X2=17.9,P＜0.01).Conclusion Imbalance in growth factors prnduction may play an important role in the process of arthritis development.Re-establishing the balance of growth factors maybe one of the mechanisms of TP in the treatment of arthritis.

This study is to explore the effects of quercetin (QUE) on the 3 week-old mice ovarian development and relative hormone levels. The 3 week-old mice were exposed to QUE (45, 25, and 5 mg x kg(-1) x hd(-1)) by gavage for 50 days. The estrous cycle during 50 days and the changes of hormone level such as FSH, LH, etc were monitored. Moreover, the ovaries were removed after sacrifice. The organ index was measured, and the ratios of different stages of follicles were analyzed by HE staining. Furthermore, the proportion of PCNA positive cells during all stages was detected by immunohistochemistry. The results showed that QUE could increase body weight of mice and reduce the anogenital distance (AGD) to some extent, and was able to disrupt mice's estrous cycle, but it could not extend or reduce the cycle regularity. It increased ovarian organ index with a dose-dependent manner. The proportion of the primordial follicle and secondary follicles rose obviously, and that of mature follicles', atretic follicles' and corpus luteums' reduced, while primordial follicle had no change. Immunohistochemistry analysis showed that QUE could effectively increase the percentage of proliferating cells in all kinds of follicles. Serum hormone assay showed that there were significant changes of FSH and LH levels. In summary, QUE showed an estrogen-like effect on mice's ovarian development. The weight of ovary, the proportion of all kinds of follicles, the development of ovarian cells and the level of plasma hormone in mice were altered obviously by oral administration of QUE.

Objective To evaluate the efficacy and feasibility of Flu/ivBu/Tl" conditioning regimen for the treatment of refractory or relapsed acute non-lymphocytic leukemia in patients receiving allogeneic hematopoietie stem cell transplantation. Methods Seven patients with refractory or relapsed acute non-lymphocytic leukemia received HLA identical peripheral blood hematopoietie stem cell transplantation (PBSCT) following Flu/ivBu/TY conditioning regimen, which consisted of fludarbine, busulfex and thiotepa. All patients received cyclos-porin A (CsA) and mycophenolet mofetil (MMF) for prophylaxis of graft - versus - host disease (GVHD). Results The Flu/IVBu/TT regimen was tolerated very well, without severe regimen related toxicity. In the 31-month median follow-up duration, 5 of 7 patients were a-live in disease-free situation. Conclusion The Flu/ivBu/TT conditioning regimen reduced transplantation-related toxicities and offered high long-term disease-free survival, and was tolerated very well. Allogeneie hematopoietie stem cell transplantation using Flu/ivBu/TT condition-ing regimen is a safe and effective option for the patients with refractory/relapsed acute non-lymphocytic leukemia.