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.

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 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 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.

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 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.

Objective To evaluate the efficacy of Flu/CTX conditioning regimen for the treatment of severe aplastic anemia in pa- tients receiving allogeneic hematopoietic stem cell transplantation. Methods Nine patients with severe aplastic anemia received HLA identi- cal peripheral blood hematopoietic stem cell transplantation (PBSCT) using Flu/CTX conditioning regimen, which consisted of fludarbine [30 mg/(m2 d) for5 days (-7 to -3) ], CTX [50mg/(kg d) for4 days(-5 to-2)]. All patients received cyclosporin A (CsA) and mycophenolet mofetil (MMF) for prophylaxis of graft-versus-host disease(GVHD). Results The Fiu/CTX regimen was very well toler- ated, with no severe regimen related toxicity. In all patients, the median days of neutrephil exceeding 0. 5×109/L and platelet exceeding 20 ×109/L were 12 days (range 10-16 days) and 16 days (range 14-19 days), respectively. Complete chimerism was achieved in all pa- tients at one month after PBSCT. Two patients had acute GVHD and one had chronic GVHD. In the 39-month median follow-up duration, all patients were alive in disease-free situation. Conclusion The Flu/CTX conditioning regimen may reduce transplantation-related toxicities and can achieve full chimerism and high long-term disease-free survival. Allogeneic hematopoietic stem cell transplantation using intravenous Fiu/CTX conditioning regimen is a safe and effective treatment method for the patients with severe aplastic anemia.

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.

Objective To explore the emergency PCI symptom-onset-to-balloon time (SOTBT) influence on the local cardial function in patients with acute ST-segment elevation myocardial infarction (STEMI) by two-dimensional speckle tracking imaging (2D-STI) technology.Methods Sixty cases were ifrst diagnosed with ST-elevation myocardial infarction (STEMI) and undergone emergency PCI surgery in Huangshi central hospital from June 2014 to June 2016. According to SOTBT, patients were divided into two groups: SOTBT≤6h myocardial infarction group, 6 h