Objective: To analyze the effects of miR-138 on the expression of small glutamine-rich TPR-containing protein A (SGTA) and cell adhesion-mediated drug resistance (CAM-DR) phenotype in non-Hodgkin's lymphoma (NHL). Methods: The adhesion model was constructed using fibronectin (FN) or bone marrow stromal cells HS-5. The effect of miR-138 on the expression of SGTA was analyzed by Western blotting and RQ-PCR. Dual-luciferase assays were performed to probe the effects of miR-138 on SGTA 3' UTR activities. Subsequently, we investigated the effect of miR-138 on cell cycle, adhesion ability and CAM-DR. Moreover, the correlation between miR-138 expression and therapeutic response was analyzed in 35 paraffin-embedded diffuse large B cell lymphoma samples. Results: Our data showed that adhesion of NHL cells to FN or HS-5 cells significantly increased miR-138 expression (P<0.05). Knockdown of miR-138 markedly increased the protein (all P<0.05) but not for mRNA (all P>0.05) levels of SGTA in NHL cell. The luciferase activity of SGTA 3' UTR was significantly suppressed by miR-138 transfected cells (0.73±0.03 vs 1.00±0.02, t=0.914, P=0.002). No change in terms of reporter activity was observed in SGTA 3'UTR mutant transfected cells (0.93±0.04 vs 1.00±0.02, t=1.375, P=0.241). Also we found that ectopic expression of miR-138 significantly induced cell cycle arrest at G(1) phase in both suspension and adherent cells (all P<0.05). Knockdown of miR-138 had no effect on cell adhesion ability (all P>0.05). More importantly, in suspension cells, knockdown of miR-138 significantly decreased the percentage of doxorubicin-induced cell death. However, knockdown of miR-138 dramatically increased the percentage of doxorubicin-induced cell death in FN/HS-5-adherent cells. Furthermore, the miR-138 expression was significantly higher in patients with progression of disease/stable disease than those experiencing complete response/partial response (9.72±1.11 vs 3.06±0.22, t=9.144, P<0.001). Conclusion: MiR-138 promoted CAM-DR phenotype through cell adhesion-mediated SGTA down-regulation and cell cycle arrest.
Carrier Proteins ; Cell Adhesion ; Cell Line, Tumor ; Cell Proliferation ; Drug Resistance, Neoplasm ; Gene Expression Regulation, Neoplastic ; Humans ; Lymphoma, Non-Hodgkin ; MicroRNAs ; Molecular Chaperones ; Phenotype

OBJECTIVETo investigate the induction of antitumor immune responses and therapeutic effects of 10-hydroxycamptothecinc-treated (HCPT) DC-Hepa fusion vaccines by DC fused with hepal-6 cell from hepatoma.

METHODSThe fused cells were isolated by magnetic cell sorting and adherent culture. Cell apoptosis was detected by Rhodamine123/PI double-labeled assay, CTL activity by 4 h (51)Cr releasing assay. Protective and therapeutic effects of the fusion vaccine to the tumor-bearing mice was also observed.

RESULTSThe apoptosis rate was 29.7%+/-4.1% when DC-Hepa fusion vaccine was treated with 50 microg/ml HCPT for 24 h. After treatment with the HCPT-DC-Hepa fusion vaccine, the tumor grew obviously slowly, survival period of the mice was prolonged, induced more potent CTL cytotoxicity, and resisted against the rechallenge of Hepal-6 cells.

CONCLUSIONVaccination with HCPT-DC-Hepa fusion vaccine could elicit potent antitumor responses, which will provide a new approach to the DC-mediated therapeutic antitumor immunity.

Animals ; Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; Camptothecin ; analogs & derivatives ; pharmacology ; Cancer Vaccines ; administration & dosage ; genetics ; immunology ; Cell Fusion ; Cytotoxicity, Immunologic ; Dendritic Cells ; immunology ; transplantation ; Female ; Liver Neoplasms ; immunology ; pathology ; Mice ; Mice, Inbred C57BL ; Neoplasm Transplantation ; Rats ; T-Lymphocytes, Cytotoxic ; immunology ; Tumor Cells, Cultured

To study the dynamic change law of bioactive constituents from Polygonum multiflorum, and to explore the optimal harvest period of P. multiflorum. Determination of stilhene glucoside, anthraquinones and catechin from P. multiflorum in different harvest times by MEKC-DAD, and principal component analysis (PCA) was used to comprehensive evaluation for bioactive constituents. There are obvious differences among the contents of active ingredients in various collecting periods samples, the content of stilbene glucoside was the highest in November, the total content of combined anthraquinone was the highest in November and December, the content of catechin was the highest in September. The comprehensive evaluation index obtained with principal component analysis showed that the sample collected in November is significantly higher than those with other samples. The optimal harvest period of P. multiflorum is November.
Electrophoresis ; Fallopia multiflora ; chemistry ; growth & development ; metabolism ; Time Factors

Objective To investigate the high-risk behaviors related to acquared immune deficiency syndrome/sexually transmitted disease (AIDS/STDs) infection among fishermen in Lv-si harbor,Jiangsu province.Methods A cross-sectional study was designed to investigate the research participants' demographic characteristics,high-risk behaviors that related to AIDS/STDs.Logistic regression was performed to measure the associations between potential risk factors and reported potential high-risk sexual behavior.Results 817 fishermen participated in the study and casual or commercial sex activities appeared to be the main high-risk behavior for AIDS/STDs infection in the target population.The rates of casual and commercial sex reported were 18.1％ and 28.9％ among fishermen.Risk factors associated with AIDS/STDs related high-risk behaviors among fishermen were high mobility (OR=1.516,P=0.038),higher lifetime sex frequency (OR=1.422,P=0.002)and unmarried status ( OR =7.527,P=0.014).Protective factors against high-risk behaviors were low intake of alcohol (OR=0.803,P=0.053),negative STD history (OR=0.268,P=0.001 ),age of initial sexual intercourse at or older than 22 years (OR =0.440,P=0.000) of age,as well as negative attitude toward multiple sexual parmers (OR=0.662,P=0.023 ) and legitimation for commercial sex (OR=0.612,P=0.007).Conclusion There were risk behaviors of AIDS/STDs in those infected fishmen.Casual and commercial sex were common high-risk behaviors.

OBJECTIVETo study the influence of different age and short or long segments of pedicle screw fixation to the clinical efficacy of early single thoracolumbar fracture.

METHODSFrom June 2005 to June 2008, 27 patients of early single thoracolumbar fracture were treated using short or long segments pedicle screw instrumentation, fracture vertebral (AO classification: type A1 or A2) was between T11 or L2. All patients were divided into A or B group according to age. A group: 12 cases mean age (32.6+/-10.7) years old (range, 16-55 years old). B group: 15 cases mean age (66.8+/-9.2) years old (range, 56-78 years old). All patients were treated with bony autograft by transpedicular of fracture vertebral and internal fixation by pedicle instrumentation. Pedicle screws were inserted in the pedicles of above and lower adjacent vertebral body of fracture vertebral, and others were inserted in the pedicles of above and lower two vertebral bodies of injured vertebral. Recorded operation time, blood loss and occurrence of complications. All patients took X radiograph plane examination (anterior-posterior position and lateral position) before operation and during 1 week of post operation and more than 1 year of follow up. Measured percentage of anterior compression vertebral high and kyphosis angle of the fracture vertebral by the same one group doctors.

RESULTSMean follow up time was (29.6+/-9.1) months (range, 10 - 34 months). The patients using short segments pedicle screw fixation in A and B group, mean operation time were (102+/-16) min and (118+/-24) min (P=0.072), mean volume of loss blood were (315+/-87) ml and (331+/-87) ml (P=0.064) respectively. The patients using long segments pedicle screw fixation in A and B group, Mean operation time were (138+/-22) min and (159+/-31) min (P=0.052), Mean volume of loss blood were (446+/-102) ml and (482+/-148) ml (P=0.055) respectively. There was no statistic different significantly between A and B group. The patients using short segments fixation, preoperative, during one week of post operation, one year of follow up, in A group the percentage of anterior compression vertebral high were 41.3+/-14.0, 5.4+/-1.0, 13.6+/-1.1, and 38.5+/-11.2, 8.3+/-2.1, 21.4+/-5.2 in B group. The patients using long segments fixation, at some time of preoperative, during one week of post operation and one year of follow up the percentage of anterior compression vertebral high were 40.8+/-11.5, 4.6+/-1.2, 8.3+/-1.0 in group A, and 44.3+/-10.2, 9.7+/-2.1, 11.2+/-3.0 in group B. In group A and B the kyphosis angle of fracture segment was 17.5 degrees+/-1.0 degrees and 16.3 degrees+/-3.1 degrees before operation, 4.2 degrees+/-1.0 degrees and 6.0 degrees+/-1.1 degrees in one week of postoperation and 11.5 degrees+/-1.0 degrees, 13.4 degrees+/-3.0 degrees in one year later postoperation. All the compression vertebral high was recovered and kyphosis was corrected significantly during one week and one year after operation (P<0.05), but there was some loss of kyphosis correction rate in follow up.

CONCLUSIONThere is better clinical efficacy of short segments pedicle instrumentation for treating early thoracolumbar fracture in the young group, but long segments fixation of pedicle instrumentation is more suitable for the older group.

Adolescent ; Adult ; Age Factors ; Aged ; Female ; Follow-Up Studies ; Fracture Fixation, Internal ; methods ; Humans ; Lumbar Vertebrae ; injuries ; Male ; Middle Aged ; Spinal Fractures ; surgery ; Thoracic Vertebrae ; injuries ; Treatment Outcome ; Young Adult

OBJECTIVETo introduce a new kind of grade system for thoracic pedicle screws placement by performing postoperative computerized tomography (CT) scanning and discussion the clinical purpose.

METHODSFour hundred and fifty thoracic pedicle screws were implanted in 64 patients with the assistance of fluoroscopy. Postoperative CT scanning was conducted to determine a grade for each screw: Part A, screw entirely contained within pedicle; Part B(1), violate lateral or upper pedicle but screw tip entirely contained within the vertebral body (VB); Part B(2), violate medial or inferior of pedicle; Part B(3), tip penetrated anterior or lateral VB; Part C, violate pedicle or VB and endangers spinal cord, nerve roots, or great vessels. Based on anatomical morphometry, thoracic vertebral were subdivided into upper (T(1 - 2)), middle (T(3 - 6)), and lower (T(7 - 12)) regions. The mean follow-up period was 25.8 months.

RESULTSThe postoperative CT scanning-documented grade were determined: Part A, 367 screws (81.6%); Part B, 78 (17.3%), B(1) 40 (8.1%); Part B(2), 23 (5.1%); Part B(3), 15 (3.4%); Part C, 5 (1.1%). There were not significant difference between upper and lower thoracic that the placement of pedicle screws in part B or C. In part C, 5 pedicle screws were all in the middle thoracic.

CONCLUSIONPostoperative CT scanning should be considered as a routine examination for evaluating thoracic pedicle screw placement.

Adolescent ; Adult ; Aged ; Bone Screws ; adverse effects ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Postoperative Complications ; prevention & control ; Retrospective Studies ; Spinal Fusion ; instrumentation ; Thoracic Vertebrae ; diagnostic imaging ; surgery ; Tomography, X-Ray Computed ; methods

OBJECTIVETo establish a stable method for obtaining large quantity of highly purified immature dendritic cells (imDCs) in vitro, and identify the morphology, function and surface markers of the cells.

METHODSCD117(+) hemopoietic stem cells (HSCs) were isolated and purified from the bone marrow of healthy C57 mice by magnetic affinity cell sorting. After cell expansion by treatment with stem cell factor (SCF) and interleukin-3 (IL-3), the HSCs were induced for directional differentiation into imDCs by treatment with GM-CSF, IL-4 and IL-10. The imDCs obtained were identified by morphological and functional observation under inverted microscope, scanning electron microscope and transmission electron microscope, followed by detection of the expressions of the surface markers using flow cytometry.

RESULTSAfter 3, 5 and 7 days of culture in the presence of SCF+IL-3, the cells were expanded by 10.34-/+1.43, 22.65-/+2.71 and 54.39-/+3.08 folds, respectively. The HSCs were successfully induced to differentiate into imDCs with phagocytotic activity. The dendrites of the imDCs were short small, and appearing spinous. The expressions of surface markers were detected from the cells showing the phenotype of CD11c(+), I-A/I-E(low), CD40(-), CD80(-), CD86(-).

CONCLUSIONThe method described allows steadily acquisition of large quanty of highly purified imDCs and of their effective identification in vitro.

Animals ; Cell Culture Techniques ; methods ; Cell Differentiation ; Cell Separation ; methods ; Cells, Cultured ; Dendritic Cells ; cytology ; Flow Cytometry ; Hematopoietic Stem Cells ; cytology ; Mice ; Mice, Inbred C57BL ; Microscopy, Electron, Scanning ; Microscopy, Electron, Transmission ; Proto-Oncogene Proteins c-kit

OBJECTIVETo assess the results of multi-technique with intact canal and scute rebuild operation for middle ear cholesteatoma.

METHODSAnalysis of 113 cases (males 60, females 53, with mean 42.7 years old), of which with double ears of 11 cases, and totally 124 ears (11 ears for one review, an ear for third time review, and totally 137 operations) was retrospectively performed. Ten cases less than 19 years old were operated from September 2001 to January 2006 for middle ear cholesteatoma as well as ossiculoplasty with intact canal and scute rebuild etc multi-technique. All cases were followed up for a mean (36.58 +/- 20.47) months.

RESULTSThere were 118 ears (95.2%) without cholesteatoma recurrence. Six ears were found cholesteatoma recurrence. Other 6 post-operation ears with drum re-perforation and hearing drop were re-operated. The pre-operation air-bone gap (ABG, average of 0.5, 1, 2 kHz) of 124 ears, with in initial 112 ears, was (33. 61 +/- 12.35) dB, while that of post-operation ABG was (13.58 +/- 9. 27) dB, by partnership t test, t = 18.35, P < 0.01. The pre-operation ABG in reviewed 12 ears at the first pre-operation was (38.83 +/- 12.43) dB, but post-operation ABG (10.38 +/- 8.99) dB, by partnership t test, t = 5.38, P = 0.00022. The ABG closure to within 20 dB was tested in 100 ears while ABG over 20 dB but air conduct (AC) in 40 dB was in 6 ears, and then as succeed was in 106 ears (85.5%). ABG closure within 10 dB was in 50 ears and though ABG over 10 dB but ABG reduced over 30 dB was in 9 ears (59/124, 47.6%) as which as best result. Bone conduct (BC) threshold was tested of 21 ears (16.93%) with raise 1-9 dB, of 42 ears (33.9%) with no change and of 61 ears (49.2%) with descend 1-28 dB. Of 10 cases less than 19 years old post-operation ABG was (8.80 +/- 5.27) dB, which indicated that all cases was as succeed.

CONCLUSIONSUsing intact canal and scute rebuild etc multi-technique a normal canal, hearing improvement and very low recurrent could be gained for middle ear cholesteatoma, especially in juvenile and children. The multi-technique should be almost no damage for hearing bone conduct.

Adolescent ; Adult ; Aged ; Child ; Cholesteatoma, Middle Ear ; surgery ; Female ; Humans ; Male ; Middle Aged ; Otologic Surgical Procedures ; methods ; Retrospective Studies ; Treatment Outcome ; Young Adult

OBJECTIVETo investigate retrospectively the clinical effects and recurrence rate of 143 cases who underwent one level discectomy and followed up more than 10 years. To evaluate the outcome of patients in groups of different operating age and extents of disc herniation, and analyse whether difference exists in each group.

METHODSThere were 143 patients (operation time from January 1996 to December 2000) including 80 males and 63 females, aged from 18 to 66 years old with an average of 37.85 years. The followed-up time was 10 to 15 years with an average of 12.7 years. Patients were divided into 3 groups depends on operating age: < 30 years old, 30 to 50 years old and > 50 years old; 87 patients who's pre-operative CT scan could be collected among 143 cases were divided into 3 groups depends on extents of disc herniation: I degree, II degrees, and III degrees. The final followed-up was obtained in 2011, to evaluate each group and the holistic clincal outcome with JOA scores and ODI scores, and observe whether there were difference between every groups; to judge the effects by patient himself with modified Macnab Criteria.

RESULTS(1) JOA scores pre-operation and final followed-up was 5.11 +/- 2.02 and 12.51 +/- 2.35 respectively; ODI scores pre-operation and final followed-up was 33.98 +/- 7.42 and 13.39 +/- 6.79 respectively. There were significant differences between pre-operative and final followed-up in JOA and ODI (P < 0.01). The excellent-good rate was 83.2% (119/143 ) according to modified Macnab Classification with recurrence rate of 6.3% at final follow-up. (2) Obvious difference was found in JOA scores in group who's age at operation less than 30 years old compared with other 2 groups at followed-up time, and no significant difference was found in JOA scores between other two groups ; no significant difference was found in ODI scores among the three groups. (3) Significant difference was found in JOA and ODI scores in group with III degrees lumbar disc herniation group compared with other 2 groups, and no statistical difference was found in clincal scores between other 2 groups.

CONCLUSION(1) Long-term followed-up of 143 cases prove mono-level lumbar discectomy is an option for disc herniation with good curative effect and lower recurrent rate, the technique should be the prior selection in dealing with patients with lumbar disc herniation. (2) 51% patients (19/37) in group under 30 years old endure persistent low back pain. (3) The long-term clinical effects in patients with severe disc protrution who underwent lumbar discectomy is worse than those patients with mild lumbar disc herniation.

Adolescent ; Adult ; Aged ; Female ; Follow-Up Studies ; Humans ; Intervertebral Disc ; surgery ; Intervertebral Disc Displacement ; surgery ; Laminectomy ; methods ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Retrospective Studies

BACKGROUNDGlobally, 180 million people suffer from diabetes mellitus. Islet transplantation is believed to be an almost ideal therapy for insulin-dependent patients. How to maintain the viability and the function of isolated human islets is a challenge in clinical practice. Sertoli cells are considered 'nurse cells' in the seminiferous tubules and have been used in cell graft protocols for neurodegenerative diseases and diabetes in many studies. Many researchers have used immature murine testes as the primarily source of Sertoli cells in islet transplantation because they are easily purified. Mature human Sertoli cells have been seldom investigated. In the present study, we developed a method for the isolation and culture of Sertoli cells derived from adult human testes, and investigated their effects on the function of allogeneic islets when they were cultured together in vitro.

METHODSAdult Sertoli cells were prepared successfully by two-step enzyme digestion with trypsin, collagenase and hyaluronidase. They were identified by morphological characteristics and their activity was determined by MTT colorimetry over a 28-day culture time in vitro. A glucose-stimulated insulin secretion test was performed to detect the effects of Sertoli cells on allogeneic islets' function when they were co-cultured for 21 days in vitro.

RESULTSIn cultured cells, mature human Sertoli cells accounted for more than 90% of total cells. The activity of Sertoli cells reached 95% and they remained highly cytoactive for a long time in vitro (P > 0.05). Compared with the islets cultured alone, the co-cultured islets with allogeneic Sertoli cells maintained higher sensitivity to glucose stimulation for the duration of the experiment (P < 0.01).

CONCLUSIONSA method of isolation and culture of Sertoli cells from adult testes has been established. Sertoli cells could enhance allogeneic islets' function when they were co-cultured in vitro. They could be a helper cell in islet transplantation.

Adult ; Cell Separation ; methods ; Cell Survival ; Cells, Cultured ; Coculture Techniques ; Humans ; Islets of Langerhans ; physiology ; Islets of Langerhans Transplantation ; Male ; Sertoli Cells ; cytology ; physiology