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 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 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 validate the feasibility and accuracy of iterative decomposition of water and fat with asymmetry and least squares estimation-quantitative fat imaging (IDEAL-IQ) in fat quantification using fat-water model. Methods A homogeneous fat-water mixture model consisting of various known fat-fractions were described, and the fat fraction was 0.00, 0.01, 0.02, 0.04, 0.06, 0.08, 0.10, 0.14, 0.18, 0.22, 0.26, 0.30 g/ml respectively. A water-vaseline separated model was also described. IDEAL-IQ was performed. Thin slices were acquired for fat-water mixture model and repeated 3 days later. Nineteen slices of 14 mm-thick parallel to the water-vaseline boundary in 1 mm steps from vaseline to water
were acquired. The fat-fractions in 11 slices of fat-water mixture model were measured on FatFrac images. Accuracy was assessed through single sample t test or Kolmogorov-Sirmov test. Measured fat-fractions of the same known fat-fraction were assessed through independent samples t test between two scan times. Linear regression was used to assess the relationship between known fat-fractions and measured fat-fractions. Slices containing the water-vaseline boundary were measured with ROI in the middle of the FatFrac images. The relationship between measured fat-fractions and locations of scanning was exploded using curve fitting. Results (1) Fat-water mixture model: no significant difference(P>0.05) was found between measured fat-fractions and known fat-fractions when it was 0.00, 0.02, 0.06 and 0.08 g/ml with the measured fat-fractions 0.60%, (2.30 ± 0.60)%, (5.76 ± 1.40)%, (7.62 ± 1.40)% respectively for the first time. No significant difference(P>0.05) was found between measured fat-fractions and known fat-fractions when it was 0.00, 0.02, 0.10 g/ml with the measured fat-fractions 0.04%, (2.32 ± 0.60)%, (9.41 ± 1.00)%respectively for the second time. Measured fat-fraction was inlinear relation with known fat-fraction:Y=0.898X+0.224, r2=0.993, P<0.01, F=36 129.548.(2) Water-vaseline separated model: measured fat-fraction increased as scanning location changed, Y=0.045X2-0.499X-4.474, r2=0.978, P<0.05, F=350.623.Conclusions IDEAL-IQ can be used to quantify fat content with good repeatability and can accurately assess the actual fat content from the linearrelationship.

Objective To study the relationship between nasopharyngeal carcinoma and positive IgA antibodies of EB virus nuclear antigen 1 (NA),EB virus capsular antigen (VCA) and EB virus Zta protein.Methods The serum EB virus VCA-IgA,NA1-IgA and Zta-IgA antibody in 17 175 cases were detected by ELISA.782 cases of EB virus antibody-positive subjects were further given nasopharyngeal CT imaging,electronic nasopharyngoscopy.Finally confirmed by biopsy and immunohistochemistry.Comparison of serology results EB antibody-positive individual,while the two antibody-positive and antibody while three positive nasopharyngeal carcinoma detection rate,combined with evaluation of EB virus antibody detection in nasopharyngeal carcinoma screening value of high-risk groups.Results 17 175 cases of medical groups,the EB virus antibodies were detected in 782 cases,with the most positive individual,accounting for 535 cases (68.41％),nasopharyngeal cancer diagnosed in 15 cases (2.80％);two antibody positive while 213 cases (27.24 ％),diagnosed 49 cases of nasopharyngeal carcinoma (23.00％);least three antibody positive while only 34 cases (4.35％),nasopharyngeal cancer diagnosed 18 patients (52.94％);by physical examination,in 16,393 cases of EB virus antibody negative population by nasopharyngoscopy and throat CT and other medical examination,the final diagnosis of nasopharyngeal carcinoma in 6 cases (0.04％).Conclusion Three EB virus antibody combined detection greatly improves the rate of nasopharyngeal cancer diagnosis;we must also do other physical examination to prevent EB virus antibody-negative nasopharyngeal crowd missed.

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 explore the prevalence of autoimmune liver disease-related antibodies in patients with PBC, and study the clinic significance of autoimmune liver disease related antibody profiles in patients with PBC. Methods The anti-AMA in 247 specimens from patients with liver disease, including 173 PBC, 37 AIH and 37 LDC were detected by IIF. Anti-AMA-M2, anti-GP210, anti-SP100, anti-SLA, anti-LCI and anti-LKM-1 antibodies were measured by ELISA. Results The positive rates of anti-AMA, anti-AMA-M2, anti-GP210, anti-SPl00, anti-LC1, anti-SLA and anti-LKM-1 antibodies were92. 5% (160/ 173), 86.7% (150/173), 35. 8% (62/173), 24. 3% (42/173), 0.6% (1/173), 0% (0/173) and 0.6%(l/173) in PBC group, 18.9% (7/37), 5.4% (2/37),8.1% (3/37),13. 5% (5/37),0% (0/ 37 ) ,5.4% (2/37 ) and 2. 7% (1/37 ) in AIH group and 5.4% (2/37), 2.7% (1/37 ) , 5.4% (2/37 ) , 10. 8% (4/37) , 0% (0/37) , 0% (0/37) and 0% (0/37) respectively in LDC group. Anti-AMA, anti-AMA-M2 and anti-GP210 was detected more frequently in patients with PBC group than AIH group (x~2 =101.3,100.8 and 11.0,P<0.01) while anti-SLA was detected more frequently in patients with AIH group than PBC group (x~2 = 9. 4, P < 0.01). The levels of ALT, TBIL, DBIL, GGT and ALP were higher in patients known to have positive anti-GP210 ( U = 1212.0,1199.0,1218.0,1074.0,1030. 0,P < 0. 01) and the levels of IgM were higher in patients known to have positive AMA ( U = 94.0, P <0.05). Conclusions Anti-LCI, anti-SLA and anti-LKM-1 antibodies in PBC and AIH are detected at a very low frequency in the corhort. Anti-GP210 antibody is found to be associated with the severity of liver damage while AMA is found to be associated with immunologic function in patients with PBC. There is little significance for screening anti-LCI, anti-SLA, anti-LKM-1 antibodies in patients with autoimmune liver diseases. It is of importance to detect anti-AMA and anti-GP210 antibodies for diagnosis of PBC.