Objective To study the clinical characteristics, diagnosis, treatment and prognosis of cancer metastasis to the thyroid gland. Methods A restropective review was performed on 35 patients with cancer metastasis to the thyroid gland in our hospital from 1958 to 2010. Diagnosis was confirmed by fineneedle aspiration cytology or histopathology in all cases. Results Primary tumor origin was identified in all but three cases. The lung was the most common primary tumor site( n = 16), followed by esophagus( n =9),breast ( n = 2), kidney ( n = 2), hypopharynx ( n= 1 ), nasopharynx ( n = 1 ) and soft palate ( n = 1 ). Thyroid metastasis was found before the diagnosis of the primary tumor was established in 12 cases; in the other 23 cases, the time lapse from diagnosis of the primary tumor to metastasis to the thyroid gland ranged from 0- 168 months, median 24 months. In 6 patients, this interval was more than 36 months. Fine-needle aspiration cytology ( FNAC ) confirmed metastatic malignancy in 7 patients, histology in 24, combined application confirmed the diagnosis in 4. After the metastasis to the thyroid gland was established, the median survival time for the entire group was 11.5 months, the 1-, 3- and 5-year survival rate was 43.8%,27. 8% and 11.9%, respectively. 28 patients were treated surgically, and 7 received nonsurgical therapy.The overall survival rate in the surgical group was higher than that in the nonsurgical group (P ＜0. 01 ). In those patients with metastatic cervical lymph nodes, median survival time did not vary from patients undergoing thyroidectomy with neck dissection to those undergoing thyroidectomy alone (P ＞ 0. 05 ).Conclusions Metastasis of carcinoma to the thyroid gland has an occult occurrence, thyroid metastases can be detected conclusively with FNAC. Metastasis to the thyroid gland implies advanced malignant tumors,often with poor prognosis.

Objective To investigate the role of IL-6 in the co-cultivation system of porcine hepatocytes with bone marrow mesenchymal stem cells (MSCs) in vitro.Methods Mononuclear cells were isolated from bone marrow aspirate of swines (n = 3) by 1.077 g/ml density gradient centrifugation.MSCs of passage 3 and primary hepatocytes harvested by a two-step in situ collagenase perfusion technique were inoculated in Millicell culture inserts at a seeding ratio of 1:2,and functional changes of heterotypic interactions were characterized.Levels of IL-6,TNF-α,and TGF-α were measured respectively.Results Hepatocyte viability was greater than 95%.Hepatocyte performance levels such as albumin secretion and urea synthesis were all significantly enhanced in co-culture group compared with hepatocyte homo-culture (P＜0.05).The IL-6 level also significantly increased in co-culture group (P ＜ 0.01).A significant decrease of hepatic functions was observed upon neutralization of IL-6 in co-culture.Conclusion MSC-derived IL-6 plays a key role in the up-regulation of hepatocyte functions in this co-culture.

Objective To establish porcine hepatocyte co-culture system with bone marrow mes-enchymal stem cells (MSCs) in vitro for the ideal cell source of bioartificial liver. Methods Mononu-clear cells were isolated from bone marrow aspirate extracted from the anterior superior lilac spine of swines (n=3) by 1.077 g/ml density gradient centrifugation. MSCs of passage 3 and primary hepato-cytes harvested by a two-step in situ collagenase perfusion technique were randomly distributed, and the morphological and functional changes of heterotypic interactions were characterized. Results The purity of the third passage MSCs and primary hepatoeytes was more than 90 % and 99%, respectively.Hepatocyte viability was greater than 95 %. A rapid attachment and self-organization of three-dimen-sional hepatocyte spheroids were encouraged in co-culture. Heterotypic junctions remained similar to that of hepatocytes in vivo. Hepatocyte performance levels such as albumin secretion and urea synthe-sis were all significantly enhanced in co-culture group compared with hepatocyte homo-culture (P<0.05). The best hepatic function levels were achieved on day 2 and moderately decreased in the following co-culture days. Conclusion Co-cultivation of porcine hepatocytes and MSCs may preserve hepatocyte morphology and function, which could contribute to the functional bioartificial liver.

ObjectiveTo study the method of cell isolation,primary culture and identification of subchondral bone cell of osteoarthritis(OA) rabbits.Methods The rabbit instable knee joint models were made by modified Hulth modeling method.The osteoblasts were harvested from the subchondral bone of rabbits by collagenase and tissue explants attachment.The morphology observation and biological identification were performed by inverted microscope and immunocytochemistry staining,respectively.The proliferative activity of cells were detected by MTT and the expression of Ⅰ-collagen at gene level was detected.ResultsThe cells started to appeared on the 11th day after the attachment.The cells form were fusiformis and triangle,the nucleolus were clear.The cultured cells had typical osteoblast morphological characteristics.The cells obtained from subchondral bone of rabbits were identified to be osteoblast by immunocytochemistry staining.The proliferative activity of cells were equably proliferation which detected by MTT.ConclusionThe modified method provides better way to obtain ideal subchondral osteoblast and the co-culture method is suitable for the study of OA microenvironment,which can simulate interactions of the subchondral osteoblast,synovial cells and chondrocyte.

ObjectiveTo identify the factors influencing the transfer of porcine endogenous retroviruses across the membrane of a new bioartificial liver (BAL),which is a necessary caution to consider for BALs carrying porcine hepatocytes.MethodsA novel porcine BAL composed of two circuits was constructed using a plasma filter with membrane pore size of 500 nm and a plasma component separator with membrane pore sizes 10 nm,20 nm,30 nm,and 35 nm.Co-cultured cells of porcine hepatocytes and mesenchymal stem cells or single porcine hepatocytes were incubated in the bioreactors.The BAL was continuously worked for 72 hours during which the supernatant was collected from the internal and external circuits every 12 hours.PERV RNA,reverse transcriptase (RT) activity,and in vitro infectivity from the supernatant were detected.ResultsIn the plasma filter group,the PERV RNAlevels were the same in both circuits,suggesting little to no effect of the plasma filter on the PERV RNA's crossing.With plasma component separators,PERV RNA was found in the external circuits at different times without positive RT activity and HEK293 cell infection,but its level was reduced significantly.The larger the membrane pore size was,the earlier and the more RNA was detected.ConclusionsThe membrane pore size,the treatment time and the viral level in the internal circuit are contributing factors influencing the transfer of PERV RNA across the membrane in a BAL.

Objective As an effective means of liver function support for acute liver failure, bioartificial liver has seen great progress in recent years. However, the development of this type of device is currently hindered by limited oxygen transport to cultured hepatocytes. In this study we try to resolve this problem by supplementing the circulating medium of the bioreactor with red blood cells.Methods Freshly isolated primary porcine hepatocytes were inoculated into our newly designed bioreactor and were divided into two groups: RPMI1640 was circulated in the control group and porcine red blood cells were added into the culture medium in the experimental group. The culture media in both groups were oxygenated through extra-corporeal membrane oxygenation, and the oxygen consumption rate (OCR) in the bioreactor was measured by a blood gas analyzer. Liver-specific functions and glucose consumption were also determined. Results The OCR of the experimental group was 1.5 fold that of the control group, and the glucose consumption rate was twice that of the control group. The liver-specific functions of the experimental group were significantly higher than those of the control group in terrns of albumin secretion and urea synthesis. Conclusion Supplementing the circulating medium of the bioreactor with red blood cells can significantly improve the oxygen supply in the bioreactor, thereby enhancing the glucose consumption and liver-specific functions of hepatocytes. This method is convenient and effective, and is expected to be an effective means to resolve the problems of oxygen supply in the bioreactor.

Objective To investigate variations in bFGF,NGF,and brain-derived neurotrophic factor(BDNF) following sciatic nerve injury in rabbits and the optimum time to perform stem cell transplantation.Methods Sixteen New Zealand rabbits were divided into control group and groups at 3,7,and 14 days postinjury according to the random number table,with 4 rabbits per group.Rabbit models of the sciatic nerve injury were induced by forceps.Structural change of the injured nerve tissues were observed with HE staining.Contents of NGF,bFGF,and BDNF in supernatants of homogenated sciatic nerves were detected by ELISA test.Results Level of bFGF increased slowly postinjury,reached the peak at day 7 (P ＜ 0.05),and then restored to the normal level at day 14 (P ＞ 0.05).Level of BDNF raised quickly postinjury,reached the peak at day 7 (P ＜ 0.05),and then restored to the normal level at day 14 (P ＞ 0.05).Level of NGF increased rapidly postinjury,reached the peak at day 3 (P ＜ 0.05),and then restored to the normal level at day 7 (P ＞ 0.05).Conclusions In the early recovery process after peripheral nerve injury,the nerve tissues regulate the secretion of NGF,bFGF,and BDNF immediately and secretions of these growth factors correlate with the time of injury.Early period (3-7 days) after injury is the best time to perform nerve repairing,nerve transplantation,and stem cell transplantation.

In this study, the rapid resolution liquid chromatography quadrupole time-of-flight mass spectrometry ( RRLC-QTOF/MS ) was used to profile the metabolites of urine samples from Childhood Pneumonia ( CP) patients and healthy controls and find the potential biomarkers which can support evidence to early diagnose and cure the disease. Choose 10 CP patients ( age 47. 72 ± 2. 35 months) and 10 healthy controls ( age 46 . 65 ± 1 . 97 months ) . The urine samples were analyzed by RRLC-QTOF/MS and then the resulting data matrices were analyzed by principal components analysis ( PCA ) to find the potential biomarkers. Urine samples of CP patients were successfully distinguished from those of healthy controls. A total of two significantly changed metabolites have been found and identified as potential biomarkers. It is suggested that the disorder of purine metabolism and amino acid metabolism may play an important role in the mechanism of CP.

Objective To replicate experimental animal model of knee osteoarthritis and to investigate the method of culture and biological characteristics of rats synoviocytes of gonarthritis in vitro.MethodsAnimal models of knee osteoarthritis were made by the Modified Hulth method.4 weeks after the replicating experiment,synovial tissues were mechanically isolated and enzyme-digested and the growth of the synovial cells was investigated.Results The synovial tissues were obviously hyperplasia in the model made by the Modified Hulth method.The synovial cells were abundant after enzyme-digested cultivation and the cell activity was higher than 98%.Conclusion The study exhibits that the Modified Hulth method apparently promotes the hyperplasia of synovial tissues.The methods of isolation and cultivation of the synovial cells in vitro is proved to be simple and feasible.

Objective To investigate the effects of different cyclic tensile strain on the proliferation of human anulus fibrosus cells from degenerated discs.Methods Anulus fibrosus(AF) cells were isolated from a degenerated human IVD,expanded in monolayer,and cyclically strained for 3 hours,applying 0,5％,10％,15％ and 20％ strains at a frequency of 0.25 Hz with the use of the DioDynamic test instrument.The flow cytometry method was used to examine the Af cells proliferation at 24 hours following application of the cyclic tensile strains.Results The proliferative index (PI) increased with the magnitude value of cyclic tensile strain except 20％ group.The most significant increase of proliferation index were found in 15％ group.Conclusion There might be some corelationships between magnitude of cyclic tensile strain and the proliferation of the degenerative AF cells.