BACKGROUND:The type B synoviocytes induced by transforming growth factor β1(TGF-β1) have the potential to differentiate into chondrocyte,which can remain the phenotype in vitro. However,whether the transfected cell combined with scaffold can form cartilage tissues need further research. OBJECTIVE:Rabbit type B synoviocytes was transfected by liposome method in vitro,combined the cells with Pluronic-F127,and then implanted into nude mouse to construct tissue engineering cartilage. Additionally,to explore the feasibility of synoviocytes differentiate into chondrocytes.DESIGN,TIME AND SETTING:The randomized animal experiment of cytology observation. The experiment was performed at the Medical Research Center,the Second Affiliated Hospital of Sun Yat-sen University between April 2007 and May 2008.MATERIALS:Healthy New Zealand white rabbits,aged 3 months,and 12 BALB/c nude mice,aged 4 weeks,weighted 20 g.METHODS:The synovial membrane tissues were taken out from the rabbit knee,isolated by enzyme digestion,and then transfected. The positive cloning was screened by G418,and the expression of TGF-β1 and collagen Ⅱ was detected. The Pluronic-F127 was dissolved at 4℃ and prepared fluid with concentration of 30%. The fluid was mixed with cells. At the same time,there were 2 groups as the control group:chondrocytes with Pluronic-F127,and synoviocytes transfeced by pcDNA3.1(+)and Pluronic-F127. The cell density was 5x1010/L. Each compound (0.2 mL) was injected into the subcutaneous of the nude mouse back. Rats were sacrificed at weeks 4,6 and 8 to harvest samples.MAIN OUTCOME MEASURES:Cell growth curve;phenotype change of the cells after transfection;histological observation of the tissue in the subcutaneous of the nude mouse back. RESULTS:Cell growth curve demonstrated that the living activity of the transfected cells was temporary decreased,which returned into normal level at days 6,7 after transfection. At day 4 after transfection,TGF β1 were positive expressed,and at day 7,the collagen Ⅱ staining were positive. The compounds in the subcutaneous of the nude mouse back formed immature chondroid tissues at week 4,which turned to mature chondroid tissue at week 8,and the collagen Ⅱ staining were positive.CONCLUSION:The transfected synoviocytes can express the phenotype of chondrocytes in vitro,and form chondrocyte-likecells. The transfected synoviocytes with Pluronic-F127 can form chondroid tissues in nude mice.

Objective To investigate the value of high-resolution 3.0T MR in the assessment of local infiltration of preoperative rectal cancer.Methods A total of 168 patients pathologically proved rectal cancer underwent both conventional pelvic and rectal high-resolution before operation, and the imaging findings were reviewed retrospectively.The accuracy of preoperative high-resolution 3.0T MR in prediction of pathological staging was assessed,and the characteristic imaging features of local infiltration in preoperative rectal cancer were discussed.Results The relationship between circumference invasion of colorectal cancer and the pathological T staging was moderately positive (rs=0.530,P=0.003).Compared the staging of colorectal cancer on MRI with pathologic T staging,the overall diagnostic accuracy was 84.52%,and there was a stronger correlation between MRI findings and pathological staging (rs=0.837,P=0.001).The best single parameters for diagnosing T3 stage rectal cancer on MRI were nodular convex of the tumor and muscular signal interruption,with 91.1% specificity and 89.7% sensitively respectively.And the best combination of parameters was the cord appearence of intestinal wall and muscular signal interruption,with 89.3% specificity and 78.0% sensitively respectively.Conclusion High-resolution 3.0T MR can be preferable to evaluating local infiltration of rectal cancer, showing a higher clinical value to asseee T staging of preoperative rectal cancer.

Objective To investigate the experimental methods of transferring the synoviocytes with the reconstructed pcDNA3.1-TGF-β1 gene by the liposomes and study the feasibility of self-induction of synoviocytes to the chondrocytes in vitro so as to provide a scientific and experimental basis for the further gene enhanced tissue engineering research in articular cartilage repair.Methods Synoviocytes were cultivate in vitro and purified to construct the eucaryotic expression plasmid carrying the recombinant rabbit TGF-β1 gene.By means of Lipofectamine 2000,the synoviocytes were transfected with pcDNA3.1-TGF-β1 (experimental group) and with pcDNA3.1 ( + ) blank plasmid (control group).The synoviocytes free from transfection was set as blank group.After 48 hours of transfection,the cells were screened by G418.Representative sections from among the positive clones were used for RT-PCR assay of instant expression of TGF-β1.The other sections were used for immunohistochemical analysis with antibodies to TGF β1.Screening of cells by G418 was continued for 12 days of cell counting and drawing the growth curve.The antigens of TGF-β1 and collagen Ⅱ were examined every week three weeks after transfection.All images were processed by using analysis instrument (Image-Pro Plus V6.0).Statistical analysis was conducted with SPSS 13.0 software package.The difference between groups was tested by using variance ( ANOVA) analysis.Results The transfection efficiency in experimental group was 18% ,with temporary decreased living activity of the transfected cells shown by growth curve.The cell population decreased to 3.6 × 104/ml four days after transfection.After 72 hours of transfection,the positive fragment of TGF-β1 was detected by RT-PCR assay,and immunohistochemical staining of antibiotics to TGF-β1 showed positive particles only in the experimental group.After three weeks of tranfection,the immunohistochemical analysis with antibodies to TGF-β1 and type Ⅱ collagen showed that there were positive particles in the transfected cells in the experimental group,with no positive particle in the control and blank groups.According to the results of Image-Pro Plus V6.0,PU value of anti-TGF-β1 was (19.04±1.26) seven days after transfection,while that of control and blank groups were (4.07 ± 0.65)and (3.23 ±0.56) respectively,with statistical difference (P<0.05).PU value of anti-type Ⅱ collagen in experimental group,control group and blank group was (13.74 ± 1.27),(4.62 ±0.56) and (3.93 ± 0.38) 14 days after transfection,with statistical difference ( P < 0.05).Conclusions Transfection of the rabbit articular synoviocytes with the reconstructed pcDNA3.1-TGF-β1 gene can be successfully accomplished by using Lipofectamine 2000 method.The transfected synoviocytes can express TGF-β1 and excrete type Ⅱ collagen,have chondrogenic potential when transfected by pcDNA3.1-TGF-β1 gene and can be used as candidate cells for repair of articular cartilage.

Objective To investigate the ability of single photon emission computed tomography (SPECT) and MRI in detecting skull-base invasion in nasopharyngeal carcinoma. Methods Sixty-one patients with nasopharyngeal carcinoma received whole body and skull-base tomography SPECT, and nasopharynx and skull-base MRI before radiotherapy. The results were double-blind compared and evaluated. Results The overall positive rates of skull-base invasion detected by SPECT and MRI were 51% and 46% (P=0.508). In paitents with headache, cranial nerve palsy or both, the rates were 83% and 86% (P=1.000) ,80% and 80% (P=1.000), 88% and 94% (P=1.000), respectively. In patients with T1+T2 and T3+T4lesions,the rates were 22% and 0(P=0.031) ,74% and 82% (P=0.250) ,repectively. In patients with N0+N1and N2+N3lesions,they were 50% and 48% (P=1.000) ,53% and 40% (P=0.500) ,respectively. The conformation rate between SPECT and MRI was 85%. Binary Logistic regression analysis showed that T stage was a risk factor for positive SPECT(χ2=4.23,P=0.040, OR=3.04). Headache tended to be a risk factor for both positive SPECT and positive MRI (χ2=3.13, P=0.077, OR=4.54;χ2=3.64,P=0.056,OR=12.00). Conclusions The detection sensitivity of SPECT in skull-base invasion in nasopharyngeal carcinoma is equivalent to that of MRI. The consistency between SPECT and MRI is good. Moreover, there is a good correlation between SPECT and symptoms, signs and stage. SPECT of skullbase tomography is necessary for patients with severe headache, negative CT and those who can not receive MRI. When SPECT result is positive,skull-base should be considered to be invaded and should be defined as gross tumor volume in radiotherapy planning.

Objective To investigate the pharmaeokinetics of levofloxacin in rat's pancreatic tissue. Methods Pancreatic tissue and blood were sampled in vivo by microdialysis simultaneously. The concentrations of levofloxacin in beth blood and tissues were measured by high performance liquid chromatography. All date were analyzed by WinNonlin software. Results The maximum concentration of free levofloxacin in blood and pancreatic tissue were (65.23 ± 12.9) μg/ml at 10min and (30.56±3.22) μg/ml at 20 min, respectively, then beth continuously decreased. Concentration of free levofloxacin in pancreatic tissue was higher than that in blood from 20min to 100min, then returned to similar level. The area under the concentration curve(AUC)of unbound levofloxacin was(2465.11±258.56)min·μg~(-1)·ml~(-1) in pancreas,and (2914.38±205.73)min·μg~(-1)·ml~(-1) in blood.Conclusions Microdialysis with reversed phase high performance liquid chromatography established in this essay could be used to determine the pharmacokinetics of levofloxacin objectively. High concentration of levofloxacin in pancreatic tissue and blood was observed.

Objectiv e To investigate the correlation between ( CAG) n repeat polymorphism of androgen receptor(AR)geneandmalebreastcancer.Methods 40casesofmalebreastcancerand40controlswerecol-lected.DNA was extracted from peripheral blood and the AR gene CAG coding exon sequences for PCR amplifica -tion,sequencing and calculated the number of CAG repeats frquency .χ2 test and Logistic regression analysis were used assess the AR gene CAG repeat length frequency affect the number of male breast cancer risk .Results There was statistically significant difference in male breast cancer cases and controls the number of CAG repeat length frequency.Man for whom the(CAG)n≥22 repeat sequence had 3.52 times risk of male breast compared (CAG)n≤22(OR=3.52,P=0.036).Conclusion AR gene CAG repeat length is a predictor of the frequency of male breast cancer risk .Longer CAG repeats can increase the risk of male breast cancer .

OBJECTIVETo establish the method for quality control of Xingnao tinctures.

METHODThe Mentholum, Flos Caryophylli and Borneoum Syntheticum in this medicine were identified by TLC method. The content of Mentholum was determined by GC. Conditions of GC were: With Naphthalene as an internal standard, packed glass column, Shimadzu 7G 2.1 mm x 3.2 m, carrier gas N2, flow rate 50 mL.min-1, column temperature. 145 degrees C, ingector and detector temperature. 180 degrees C, injection way of splitless and injection volume 3 microL.

RESULTThe spots on TLC plates were clear, with no interference in the blank reference. The precision and repeatability of GC were are good respectively. The average recovery was 98.4%, RSD = 0.94%.

CONCLUSIONThis method was fast, simple, accurate and suitable for quality control of Xingnao tinctures.

Bornanes ; chemistry ; Chromatography, Gas ; Chromatography, Thin Layer ; Drug Combinations ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; isolation & purification ; Flowers ; chemistry ; Materia Medica ; administration & dosage ; chemistry ; isolation & purification ; Menthol ; analysis ; Plants, Medicinal ; chemistry ; Quality Control ; Syzygium ; chemistry

Objective To explore the role of stromal interaction molecule 1(STIMI)on prohteration and intra- cellular Ca~(2+)change in vascular smooth muscle ceils(VSMC).Methods Rat VSMC were isolated from SD rats and primary cultured.Ad-si/rSTIM1 and Ad-hSTIM1 were transfected into VSMC.The protein of STIM1 was measured by Western blot,the proliferation of VSMC was analyzed by ~3H-thymidine(~3H-TdR)incorporation and cell count,the intracellular Ca~(2+)change was assessed By Aquaeosmos system.Ruselts Fourty-eight hours after transfection,as compared with Ad-hSTIMI group,the Ad-si/rSTIMI VSMC had lower expression of STIM1 protein (P

Objective To analyze the characteristics of preoperative CTO lesions by coronary CT angiography (CCTA) and to compare the lesion characteristics and clinical data of patients with subsequent vs failed PCI.Methods A total of 113 patients were randomly selected and 116 vessels were analyzed by CCTA before PCI.The patients were further investigated as PCI success group vs PCI failure group according to their PCI result.Multivariate logistic regression analysis was used to determine the factors that affected the success of CTO intervention.The ROC curve was used to determine and evaluate the CT-CTO score and J-CTO score for diagnostic efficacy.Results The success rate of PCI was 55.2％.64 lesions were successfully opened,with the success rate of 72.4％.The prevalence of smoking in patients in the PCI failure group was significantly higher than that in PCI success group (65.4％ vs.42.2％,P ＜ 0.05).There were no significant differences between the two groups in age,gender,history of hypertension,diabetes mellitus,and myocardial infarction(P ＞ 0.05).Statistical differences were observed between the PCI success group and the PCI failure group in the presence of occlusion segment head-end bifurcation,occlusion severe incision,severe calcification (calcification ≥ 180°),occlusion segment length ≥ 20 mm,occlusion of calcification lesions,occlusion segment distal shape of the unambiguous of fiber cap shape of the distal occlusion segment under CCTA(P ＜ 0.05).In the PCI failure group,approximately 17.3％ of the patients had previous attempt to open the CTO lesions,which were higher than the PCI success group (9.4％).However,The difference was not statistically significant (P ＞ 0.05).Multivariate logistic regression analysis showed that the unambiguous distal fibrous cap of the occlusion segment and the occlusion of the proximal branch and the occlusion length ≥20 mm were the main factors affecting the failure of CTO intervention.In terms of prediction,the predictive value 30 CT-CTO score yielded a higher area under the ROC curve than that of the J-CTO score (0.8776 vs 0.7387,P ≤ 0.05).Conclusion CT angiography can predict the success rate of intervention for CTO lesions.Compared with J-CTO score,CT-CTO score has a higher predictive value.Unambiguous fiber cap shape,occlusion segment head end bifurcation,occlusion segment length ≥20 mm were the independent risk factors that affecting the success of CTO operation.

Objective To establish qualitative and quantitative methods for analyzing monotropein in Liquidambaris Radix. Methods Monotropein in Liquidambaris Radix was identified by using TLC. The contents of monotropein in the samples were determined by HPLC on an Sino Chrom ODS-BP column (4.6 mm × 250 mm, 5 μm) with methanol-0.1％ phosphoric acid solution (1:99) as the mobile phase at the flow rate of 1.0 m L/min; the detection wavelength was 235 nm; the column temperature was 25 ℃; the injection volume was 10 μL. Results Monotropein in Liquidambaris Radix could be identified by TLC. Monotropein showed good linear relation in the range of 0.150 1– 1.501 0 μg (r=0.999 8), and the average recovery rate was 95.93％ (RSD=0.60％). The contents of monotropein in Liquidambaris Radix from 10 different producing areas were among the range of 0.20％–1.15％. Conclusion The method is simple, stable, reliable and reproducible, which can be used for the quality control of Liquidambaris Radix.