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 biokinetics of 131 I and its absorbed dose in patients with metastatic DTC. Methods Forty?one patients with metastatic DTC who were referred to 131 I therapy were prospectively recruited from September 2009 to September 2011. Scintigraphic images were serially acquired after the ingestion of 131I (3.70-9.25 GBq) to assess the biokinetics. CT was used to measure the masses of the DTC lesions. Calculation of absorbed doses to DTC lesions was based on the Medical Internal Radiation Dosimetry formula of the Society of Nuclear Medicine and Molecular Imaging. Kruskal?Wallis rank sum test was used to compare the differences of absorbed dose among different tumor deposits. Results In total, 59 io?dine?avid metastatic lesions (35 in cervical lymph node, 17 in lung and 7 in bone) in 41 patients were evalu?ated and the mean absorbed dose of DTC lesions was (10.4±6.5) Gy/GBq, with range of 0.1 to 136.2 Gy/GBq. The median absorbed dose was 11.3 Gy/GBq for lymph node metastase, 10.1 Gy/GBq for lung metastases and 9.6 Gy/GBq for bone metastases. There were no statistically significant differences in absorbed doses a?mong the metastases of lymph node, lung and bone ( Z=0.907, P>0.05) . Conclusion A wide inter?lesion variation of absorbed dose to metastatic DTC is observed.

Objective To evaluate the changes in the number and activities of endothelial progenitor cells (EPCs) from peripheral blood in patients with primary Budd-Chiari syndrome (BCS),and to explore the possible mechanisms of BCS.Methods Eighty-two patients with BCS and 20 healthy subjects used as controls were recruited for this study.The EPCs from peripheral blood were counted by flow cytometry for CD34,CDl33 and KDR for positivity.The peripheral blood mononuclear cells were isolated by density gradient centrifugation and cultured for 7 days.Characterization of EPCs as adherent cells was done using double staining of FITC-UEA-1 and DiI-Ac-LDL binding.The proliferation,adhesion and migration activities were assayed by MTT chromatometry,adhesion activity assay and Transwell assay,respectively.Results EPCs (CD34+/CD133 +/KDR+) were depleted in the BCS patients as compared to the healthy controls [(0.020 ± 0.005) ％ vs (0.038 ± 0.007) ％].The proliferation activities (0.20 ± 0.04 vs 0.58 ± 0.07),adhesion activities (15.8 ± 1.6 vs 35.0 ± 2.5) and migration activities (16.1 ± 1.7 vs 23.9 ± 2.6) were significantly lower in the BCS group than the control group (P ＜ 0.05).Conclusion EPCs from the peripheral blood in patients with BCS exhibited reduced numbers and impaired proliferation,adhesion and migration activity,which may be the key factors for vasculopathy formation in primary BCS patients.

10.3969/j.issn.2095-4344.2013.26.013

Objective To investigate the relationship between the absorbed dose to the thyroid remnant and the outcome of m I ablation in patients after operation for DTC.Methods Seventy-two patients (14 males,58 females; age range:16-67 years,average age:(41±16) years) with DTC,prepared for thyroid remnant ablation,were prospectively recruited from September 2009 to September 2011.Scintigraphic images of the head and neck were serially acquired after oral administration of 3.7 GBq of 131I to assess the biokinetics of this tracer in thyroid remnant.Ultrasound was used to measure the mass of the thyroid remnant.Calculation of absorbed dose to thyroid remnant was based on the Medical Internal Radiation Dosimetry formula of the Society of Nuclear Medicine.The outcome of 131 I ablation was evaluated on the basis of stimulated Tg measurements and neck ultrasound at 6-9 months after ablation.Patients with stimulated Tg levels less than 1 μg/L and with no detectable thyroid tissue on neck ultrasound were considered successful ablation of thyroid remnant.Two-sample t test was used to analyze the data.Results The 131I uptake ratios at 24 h were 0.9％-6.3％ in 72 patients and the effective half-lives of 131I were 12.0-146.4 h.The thyroid remnant masses were 1.0-6.9 g; absorbed doses were 23-2 197 Gy,and absorbed dose rates at 24 h were 0.5-8.1 Gy/h.No significant difference was observed in absorbed doses to thyroid remnants in 43 patients having successful ablation and 29 patients having unsuccessful ablation ((363± 148) Gy vs (341± 167) Gy,t =15.097,P＞0.05).However,the absorbed dose-rates of thyroid remnants at 24 h were significantly higher in patients with successful ablation than those in patients with unsuccessful ablation ((3.7±2.1) Gy/h vs (2.9±1.6) Gy/h,t=7.908,P＜0.05).Conclusion A successful ablation is strongly dependent on the absorbed dose-rate of thyroid remnant.

Objective To determine the activities of 131I for treating differentiated thyroid carcinoma with diffuse pulmonary metastases ( DTC-DPM ) from the perspective of internal radiation dosimetry.Methods According to Medical Internal Radiation Dosimetry (MIRD) schema, the activity constraint,from which the whole bdy retention at 48 h should not exceed 2.96 GBq (2.96 GBq rule), was converted to dose-rate constraint(DRC) to lungs at 48 h ( DRCLU ·48 h ) in 131I therapy for DTC-DPM. Based on the assumption of DRCLU·48 h at 48 h in lung, the fractions of whole body activities ( F48 ), the effective half times of 131I in lungs ( TLL ) and the remainder of body ( TRB ) were 0.6-0.9, 20- 120 h, and 10- 20 h, respectively. The maximum safe activities of 131I for different human phantoms from the Organ Level Internal Dose Assessment (OLINDA) software were calculated. Results According to MIRD schema and 2.96 GBq rule, DRCLU ·48 h should not exceed 46.4 mGy/h in 131I therapy for DTC-DPM. Depending on varying F48 h,TLL and TRB, the maximum safe activities of 131I were 6.77-81.36, 5.29-56.20, 5.08-55.19 and 3.87-40. 52 GBq for the male adult, female adult, 15-year-old, and 10-year-old patients with DTC-DPM, respec tively. Conclusion Dosimetry-guided 131I therapy for DTC-DPM considers adequately the differences of 131I kinetics in individual patients and can adjust administered activities of 131I on the precondition of avoiding radiological pneumonitis and pulmonary fibrosis.

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 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.

Objective To investigate the efficacy and complications of therapeutic ERCP for patients over 80 years. Methods Data of 80 patients aging over 80 years, who tnderwent therapeutic ERCP from August 2007 to August 2009 were retrospectively studied. The etiology, complications and therapeutic effects were evaluated. Results A total of 51 patients (63. 8% ) were accompanied by cardiovascular diseases, and 43 patients were diagnosed as having malignant bile duct stricture (53. 8% ). The overail therapeutic efficacy was 96. 3%. The incidence of post-operative complications was 12. 5% (10/80). Occurrence of complications in patients with malignant tumors ( 18. 6% ) was significantly higher than that in patients with bile duct stones (5.6% ,P ＜0. 05). Incidence of complications in those with cardiopulmonary diseases was similar with that in patients without (11.8% vs. 13.8%,P＞0.05). Conclusion Age more than 80should not be considered as contraindication of ERCP. Therapeutic ERCP is effective in senior patients. Patients with advanced ages shows a relatively high incidence of complication after ERCP, which may not be related to other accompanying diseases.

For patients with DTC,the side effect of 131I therapy on fertility and offspring is an important issue since genital tissues are highly sensitive to radiation.Exposure to 131 I radiation may result in transient impairment of gonadal function in male patients caused by elevated levels of serum follicle stimulating hormone and luteinizing hormone,low sperm count and motility.In female patients,exposure to 131I radiation may be complicated with delay of menstruation,oligomenorrhea and transient cessation of menstrual period.Most of these symptoms may resolve within one year after 131 I therapy.A slightly earlier menopause is the only reported long-term side effect of 131I therapy on ovarian function.Currently available data do not indicate that exposure to 131I may cause permanent infertility in male patients with DTC.For female patients with DTC,131 I therapy does not affect fertility or pregnancy outcomes beyond one year.