BACKGROUNDBioreactors are pivotal tools for generating mechanical stimulation in functional tissue engineering study. This study aimed to create a bioreactor that can simulate urinary bladder mechanical properties, and to investigate the effects of a mechanically stimulated culture on urothelial cells and bladder smooth muscle cells.

METHODSWe designed a bioreactor to simulate the mechanical properties of bladder. A pressure-record system was used to evaluate the mechanical properties of the bioreactor by measuring the pressure in culture chambers. To test the biocompatibility of the bioreactor, viabilities of urothelial cells and smooth muscle cells cultured in the bioreactor under static and mechanically changed conditions were measured after 7-day culture. To evaluate the effect of mechanical stimulations on the vital cells, urethral cells and smooth muscle cells were cultured in the simulated mechanical conditions. After that, the viability and the distribution pattern of the cells were observed and compared with cells cultured in non-mechanical stimulated condition.

RESULTSThe bioreactor system successfully generated waveforms similar to the intended programmed model while maintaining a cell-seeded elastic membrane between the chambers. There were no differences between viabilities of urothelial cells ((91.90 ± 1.22)% vs. (93.14 ± 1.78)%, P > 0.05) and bladder smooth muscle cells ((93.41 ± 1.49)% vs. (92.61 ± 1.34)%, P > 0.05). The viability of cells and tissue structure observation after cultured in simulated condition showed that mechanical stimulation was the only factor affected cells in the bioreactor and improved the arrangement of cells on silastic membrane.

CONCLUSIONSThis bioreactor can effectively simulate the physiological and mechanical properties of the bladder. Mechanical stimulation is the only factor that affected the viability of cells cultured in the bioreactor. The bioreactor can change the growth behavior of urothelial cells and bladder smooth muscle cells, resulting in the cells undergoing adaptive changes in mechanically-stimulated environment.

Bioreactors ; Cell Line ; Humans ; Myocytes, Smooth Muscle ; cytology ; Tissue Engineering ; methods ; Urinary Bladder ; cytology ; Urothelium ; cytology

OBJECTIVETo evaluate the clinical outcome of arthroscopically assisted combined anterior and posterior cruciate ligament (ACL/PCL) reconstructions using Achilles tendon-bone allografts.

METHODSAssociated meniscus injuries were treated according to established methods prior to ligament reconstructions during arthroscopic surgery. Thirty Achilles tendon-bone allografts were used to reconstruct torn ACL and PCL in 15 knees. At postoperative follow-up, all knees were graded using the modified IKDC and the Lysholm scoring systems just as done preoperatively.

RESULTSwere analyzed compared with the contralateral healthy knees. Results: Eleven men and 4 women with a minimum of 3-year follow-up (mean 38 months) were included in the study. Preoperatively, the group ratings by the modified IKDC standards were all severely abnormal. Twelve bicruciate reconstructions were performed in subacute or chronic stage (larger than 3-8 weeks), 3 for acute ligamentous deficiencies (less than or equal to 3 weeks). The noticeable early complication was transitory local fever combined with joint effusion in one case. At postoperative follow-up, 9 knees were normal, 5 nearly normal and 1 abnormal. On Lysholm score the difference was statistically significant (t- test, P less than 0.001) before and after operation.

CONCLUSIONSAchilles tendon-bone allograft offers an alternative for simultaneous arthroscopic ACL/PCL reconstructions. However, further investigation is needed to eradicate its potential immunogenicity for better use.

Achilles Tendon ; transplantation ; Anterior Cruciate Ligament ; surgery ; Arthroscopy ; methods ; Bone Transplantation ; methods ; Female ; Humans ; Knee Injuries ; surgery ; Male ; Posterior Cruciate Ligament ; surgery ; Range of Motion, Articular ; Reconstructive Surgical Procedures ; methods ; Transplantation, Homologous

OBJECTIVEFrequency, type and clinical implications on protease and reverse transcriptase drug resistance mutations were investigated and phylogenetic analysis in antiretroviral drug-naïve AIDS patients was carried out in Henan province.

METHODS45 plasma samples were separated from the anticoagulatory whole blood, from which reverse transcription-polymerase chain reaction technique was used to amplify the partial pol gene. The sequences were analysed for genotypic antiretroviral resistance and phylogenetic relation through landing the websites http://hivdb.stanford.edu and http://hiv-web.lanl.gov, under BioEdit and DNAClub software.

RESULTSPartial pol sequences of 36 samples were successfully amplified. The major mutation rate of resistance to protease was 8.3% (3/36), including types D30A, V32A, G73C and V82A. Minor mutation rate of resistance was 100%, including types of L63PS (36/36), I93L (35/36), V77IL (34/36), A71IVT (10/36) and D60E (2/36). The mutation rate of resistance to reverse transcriptase was 38.9% (14/36). Mutation-scoring and clinical implication clewed drug resistance rates were 5.6% (2/36) and 22.2% (8/36) to protease inhibitors and reverse transcriptase inhibitors respectively, while 1 sample was potentially low-level resistant to all of the protease inhibitors and 3 samples to part of the reverse transcriptase inhibitors. Phylogenetic analysis revealed that the pol gene of 36 samples were highly homologous and having a near relative to B.US.83.RF ACC M17451. 36 samples seemed to have the same infection source while their resistance mutations were not due to drug-resistant virus infection but to the evolving of virus in vivo.

CONCLUSIONMost of the antiretroviral drug-naïve AIDS patients in Henan province were sensitive to the currently available antiviral medicine, but antiviral treatment must be in accordance with the strict procedure and to keep better adherence, to avoid the epidemics caused by drug-resistant virus.

Acquired Immunodeficiency Syndrome ; genetics ; Adult ; Anti-HIV Agents ; pharmacology ; China ; Drug Resistance, Viral ; genetics ; Female ; Genes, pol ; genetics ; Genotype ; HIV Protease ; genetics ; HIV Protease Inhibitors ; pharmacology ; Humans ; Male ; Mutation ; Phylogeny ; RNA-Directed DNA Polymerase ; genetics ; Reverse Transcriptase Inhibitors ; pharmacology

There is a dearth of case reports describing simultaneous bilateral patellar tendon ruptures in the medical literature. These ruptures are often associated with systemic disorders such as lupus erythematosus or chronic steroid use. The author describes a case of a 24-year-old man who sustained traumatic bilateral patellar tendon ruptures without any history of systemic disease or steroidal medication. We repaired and reattached the ruptured tendons to the patella and augmented our procedure with allogeneic tendon followed by wire loop reinforcement. One year after operation, the patient regained a satisfactory range of motion of both knees with good quadriceps strength and no extensor lag. The recurrent microtrauma from a history of intense sports activity and a high body mass index may have played an important role in this trauma event.
Humans ; Knee Injuries ; Patella ; injuries ; Patellar Ligament ; Rupture ; Tendon Injuries ; surgery

OBJECTIVETo study the feasibility of regenerating a whole menisci using poly-(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) scaffolds loaded with meniscal cells in rabbits undergoing total meniscectomy, and to explore its protective effect on cartilage degeneration.

METHODSA solvent casting and particulate leaching technique was employed to fabricate biodegradable PHBV scaffolds into a meniscal shape. The proliferated meniscal cells were seeded onto the polymer scaffolds, transplanted into rabbit knee joints whose lateral menisci had been removed. Eight to 18 weeks after transplantation, the rege- nerated neomenisci were evaluated by gross and histological observations. Cartilage degeneration was assessed by Mankin score.

RESULTSEighteen weeks after transplantation, the implants formed neomenisci. Hematoxylin and eosin (HE) staining of the neomenisci sections revealed regeneration of fibrocartilage. Type I collagen in the neomenisci was also proved similar to normal meniscal tissue by immunohistochemical analysis and Sirius scarlet trinitrophenol staining. Articular cartilage degeneration was observed 8 weeks after implantation. It was less severe as compared with that in total meniscectomy controls and no further degeneration was observed at 18 weeks. At that time, the regenerated neomenisci strongly resembled normal meniscal fibrocartilage in gross and histological appearance, and its mechanical property was also close to that of normal meniscus.

CONCLUSIONSThe present study demonstrates the feasibility of tissue-engineering a whole meniscal structure in total meniscectomy rabbit models using biodegradable PHBV scaffolds together with cultured allogeneic meniscal cells. Cartilage degeneration is decreased. But long-term in vivo investigations on the histological structure and cartilage degeneration of the neomenisci regenerated by this method are still necessary to determine the clinical potential of this tissue engineering avenue.

Animals ; Cartilage, Articular ; Cells, Cultured ; Knee Joint ; Menisci, Tibial ; Polymers ; Regeneration ; Tissue Engineering

OBJECTIVETo detect the factors relevant to stenosis of tracheal graft and to find feasible methods to solve this problem.

METHODSSixteen mongrel dogs were divided into groups A and B randomly and equally. Five-ring-length tracheal segments were allotransplanted. All grafts and anastomotic sites were covered with omental pedicles. In group A, no immunosuppressant was given and in group B, the recipients were treated with cyclosporine. The animals were sacrificed 4 weeks after operation, and their postmortem specimens were examined grossly and histologically. All allografts were assessed by percent patency. Epithelial regeneration and morphology of the cartilage were semiquantitatively evaluated.

RESULTSStructural integrity of the allografts were maintained better in group B than in group A. Tracheal stenosis was found to be more serious in group A. The scores of epithelial regeneration and cartilage morphology were higher in group B than in group A, and in each group positive correlation was found between the percent patency and the score of epithelial regeneration or cartilage morphology.

CONCLUSIONSImmunosuppressive drugs are necessary to maintain the structure of allografts. Tracheal stenosis is correlated closely with epithelial regeneration and morphological maintenance of the cartilage.

Animals ; Dogs ; Immunosuppressive Agents ; pharmacology ; Male ; Trachea ; pathology ; transplantation ; Transplantation, Homologous

OBJECTIVETo investigate the safety and feasibility of laparoscopic D2 gastrectomy for advanced gastric cancer.

METHODSThe clinical data of 210 cases of laparoscopic gastrectomy and 180 cases of open gastrectomy for radical (D2) gastrectomy from May 2007 to Dec 2010 were analyzed retrospectively.

RESULTSA total of 206 cases underwent laparoscopic-assisted surgery with 4 conversions. Compared to the open group, the laparoscopic group was associated with less bleeding [(208±38) ml vs. (300±52) ml, P<0.05], quicker postoperative recovery of bowel function [(2.9±0.7) d vs. (3.9±1.8) d, P<0.05], shorter postoperative length of hospital stay[(12.8±6.2) d vs. (15.6±6.8) d, P<0.05], longer operative time [(258±42) min vs. (193±30) min, P<0.05]. The number of lymph node harvested was 20.5±1.9 in the laparoscopic group and 25.8±1.5 in the open group, and the postoperative complication rate was 8.1% (17/201) vs. 8.5% (15/180), and differences were not statistically significant (both P>0.05). The recurrence rate was 2.9% (6/210) and 2.8% (5/180), and the 3-year overall survival rate was 35.6% and 37.8%, the differences were not statistically significant (both P>0.05).

CONCLUSIONSLaparoscopic radical gastrectomy for gastric cancer is safe and effective, which can reach the same range of lymph node dissection as open gastric cancer surgery and similar survival rate.

Aged ; Female ; Follow-Up Studies ; Humans ; Laparoscopy ; Laparotomy ; Lymph Node Excision ; Male ; Middle Aged ; Retrospective Studies ; Stomach Neoplasms ; surgery ; Survival Rate

OBJECTIVETo compare the clinical results of additional screws fixation on fractured vertebrae versus only short-segment posterior transpedicular instrumentation for A3 thoracolumbar fracture without neurologic deficit.

METHODSClinical data of 52 cases of thoracolumbar burst fracture without neurologic deficit were retrospectively analyzed. All patients were divided into 2 groups due to different instrumentation and all fractures were classified as type A3 according to AO Classification.From January 2005 to December 2006, 23 cases in group A were treated by short-segment posterior instrumentation combined with additional screws fixation on fractured vertebrae. There were 18 male and 5 female with a mean age of (35.3+/-8.3) years. The fracture segment included 1 in T11, 9 in T12, 11 in L1 and 2 in L2. From January 1999 to December 2004, 29 cases in group B were treated only by conventional short-segment posterior transpedicular instrumentation. There were 20 male and 9 female with a mean age of (37.3+/-6.8) years. The fracture segment included 1 in T11, 7 in T12, 20 in L1 and 1 in L2. The clinical effect and radiographic measurements were respectively compared preoperatively, immediate and 2 years postoperatively.

RESULTSAll patients were followed up and the mean follow-up time was (37.4+/-10.9) months (from 24 to 48 months). There was no statistic difference of mean JOA and VAS score between 2 groups preoperatively, immediate and 2 years postoperatively (P>0.05). The average immediate postoperative correction of Cobb's angle was 13.7 degrees+/-7.7 degrees in group A, which was statistically significantly higher than that of 8.8 degrees+/-5.0 degrees in group B (P<0.01). The mean kyphosis correction loss of 2.9 degrees+/-1.5 degrees in group A was statistically significantly lower than that of 5.0 degrees+/-2.9 degrees in group B 2 years postoperatively (P<0.01). The average restoration of anterior height of fractured vertebral body immediate postoperatively was (29.4+/-6.0)% and (21.7+/-6.9)% respectively. The mean correction loss of anterior height 2 years postoperatively was (3.1+/-0.8)% and (6.6+/-3.0)% respectively. The average restoration of posterior height of fractured vertebral body immediate postoperatively was (8.5+/-3.2)% and (6.1+/-1.8)% respectively. The mean correction loss of posterior height 2 years postoperatively was (2.0+/-0.8)% and (3.4+/-1.0)% respectively. There were significant differences in average restoration of anterior/posterior height immediate postoperatively and correction loss of anterior/posterior height 2 years postoperatively between the 2 groups (P<0.01). According to fracture fragments protruded into the spinal canal on immediate postoperative CT image, there were complete reduction in 11 cases (47.8%) and partial reduction in 12 cases (52.2%) in group A, which was statistically significantly better than those in group B (P<0.01). There was no severe neurologic complications and no other complications related to additional screws fixation postoperatively. Pedicle screw breakage occurred in 2 cases in group B and none in group A.

CONCLUSIONSBetter initial kyphosis correction and less loss of correction 2 years after operation can be obtained by using additional screws fixation on fractured vertebra for thoracolumbar A3 fracture without neurologic deficit.

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

BACKGROUNDNatural articular cartilage has a limited capacity for spontaneous regeneration. Controlled release of transforming growth factor-beta1 (TGF-beta1) to cartilage defects can enhance chondrogenesis. In this study, we assessed the feasibility of using biodegradable chitosan microspheres as carriers for controlled TGF-beta1 delivery and the effect of released TGF-beta1 on the chondrogenic potential of chondrocytes.

METHODSChitosan scaffolds and chitosan microspheres loaded with TGF-beta1 were prepared by the freeze-drying and the emulsion-crosslinking method respectively. In vitro drug release kinetics, as measured by enzyme-linked immunosorbent assay, was monitored for 7 days. Lysozyme degradation was performed for 4 weeks to detect in vitro degradability of the scaffolds and the microspheres. Rabbit chondrocytes were seeded on the scaffolds containing TGF-beta1 microspheres and incubated in vitro for 3 weeks. Histological examination and type II collagen immunohistochemical staining was performed to evaluate the effects of released TGF-beta1 on cell adhesivity, proliferation and synthesis of the extracellular matrix.

RESULTSTGF-beta1 was encapsulated into chitosan microspheres and the encapsulation efficiency of TGF-beta1 was high (90.1%). During 4 weeks of incubation in lysozyme solution for in vitro degradation, the mass of both the scaffolds and the microspheres decreased continuously and significant morphological changes was noticed. From the release experiments, it was found that TGF-beta1 could be released from the microspheres in a multiphasic fashion including an initial burst phase, a slow linear release phase and a plateau phase. The release amount of TGF-beta1 was 37.4%, 50.7%, 61.3%, and 63.5% for 1, 3, 5, and 7 days respectively. At 21 days after cultivation, type II collagen immunohistochemical staining was performed. The mean percentage of positive cells for collagen type II in control group (32.7% +/- 10.4%) was significantly lower than that in the controlled TGF-beta1 release group (92.4% +/- 4.8%, P < 0.05). Both the proliferation rate and production of collagen type II in the transforming growth factor-beta1 microsphere incorporated scaffolds were significantly higher than those in the scaffolds without microspheres, indicating that the activity of TGF-beta1 was retained during microsphere fabrication and after growth factor release.

CONCLUSIONChitosan microspheres can serve as delivery vehicles for controlled release of TGF-beta1, and the released growth factor can augment chondrocytes proliferation and synthesis of extracellular matrix. Chitosan scaffolds incorporated with chitosan microspheres loaded with TGF-beta1 possess a promising potential to be applied for controlled cytokine delivery and cartilage tissue engineering.

Animals ; Cartilage ; metabolism ; Cell Proliferation ; Chitosan ; administration & dosage ; Chondrocytes ; cytology ; Drug Carriers ; Microspheres ; Rabbits ; Tissue Engineering ; methods ; Transforming Growth Factor beta1 ; administration & dosage ; chemistry

BACKGROUNDUsing magnetic resonance imaging, diagnosis of malignant meningioma from benign meningioma with atypical features is uncertain. We evaluated the value of lipid signal in differentiating intracranial meningiomas.

METHODS1H-magnetic resonance spectroscopy (MRS) using a point resolved spectroscopy (TR/TE 1000/144 ms) sequences were performed on 34 patients on a 3.0 T scanner. Lipid peak located at 1.3 ppm was evaluated. MRS data from these tumours were compared with histopathological findings (including hematoxylin and eosin staining and KP-1 staining).

RESULTSTwenty-nine meningiomas were histologically benign (eleven meningothelial, thirteen fibrous, four transitional and one microcystic), three were atypical, and two were anaplastic. Lipid signal was detected in ten cases: two anaplastic, three atypical, two fibrous and three meningothelial meningiomas. All voxels with lipid peak in the spectrum from the tumour were evaluated. With creatinine peak in the normal white matter chosen as internal standard, lipid/creatinine ratios of anaplastic, atypical and benign meningiomas were 0.844 +/- 0.027 (range from 0.725 to 0.994), 0.465 +/- 0.023 (range from 0.239 to 0.724), and 0.373 +/- 0.016 (range from 0.172 to 0.571) respectively. Highly significant differences were noted between anaplastic and the other two subtypes. Patchy necrosis was observed in anaplastic meningioma, while focal necrosis was noted in atypical meningioma with HE stain. However, no necrosis was found in benign group. KP-1 stain demonstrated histocytes containing lipids in the necrotic region of anaplastic and atypical meningioma.

CONCLUSIONThe lipid signal at 1.3 ppm is a useful marker in evaluating the malignancy of intracranial meningiomas, especially in the differential diagnosis of anaplastic meningioma.

Adult ; Female ; Humans ; Magnetic Resonance Imaging ; methods ; Magnetic Resonance Spectroscopy ; methods ; Male ; Meningeal Neoplasms ; diagnosis ; Meningioma ; diagnosis ; Middle Aged ; Reproducibility of Results ; Sensitivity and Specificity