OBJECTIVETo gain stable genetic modification of neural stem cells (NSC) that constitutively secrete brain-derived neurotropic factor (BDNF) and to explore the biological role on the survival and neurite outgrowth of cultured dorsal root ganglion (DRG) neurons.

METHODSBDNF gene fragment from human brain cDNA libraries was obtained by using PCR. With molecular cloning technique, the recombinant stem cell viral vector with report gene was constructed, which is that MSCV-BDNF-IRES(2)-EGFP vector could encode Polycistronic mRNA. Viral particle was packaged by PT67 cell line and infected neural stem cells (mouse clone: C17.2). After selection with cloning cylinder, the expression of BDNF was assessed by immunohistochemistry enzyme linked immunosorbent assay (ELISA) and reverse transcriptase polymerase chain reaction (RT-PCR). The effects of stable gene-modified neural stem cells on embryonic mouse DRG neurons were evaluated in a dual-chambered cocultivation system at 3th, 10th day.

RESULTSRT-PCR analysis demonstrated expression of mRNA for human-BDNF. ELISA confirmed the presence of secreted BDNF 24 h after transfection and showed that the level of BDNF production by NSC-BDNF transfected C17.2 was at a rate of (14.6 +/- 0.8) ng x d(-1) x 10(-6) cells even after 3 months. With immunohistochemical analysis, compared with the control, the longer neurite outgrowth of cultured DRG cells and the more survival neurons were observed in NSC-BDNF transfected cells group.

CONCLUSIONSBDNF gene could be stably expressed in C17.2 cell line by MSCV, and the BDNF gene-modified NSC markedly enhances the survival and neurite outgrowth of cultured DRG neurons.

Animals ; Brain-Derived Neurotrophic Factor ; biosynthesis ; genetics ; Cell Culture Techniques ; Cell Survival ; Cells, Cultured ; Coculture Techniques ; Ganglia, Spinal ; cytology ; Humans ; Mice ; Neurons ; cytology ; metabolism ; Rats ; Rats, Sprague-Dawley ; Stem Cells ; cytology ; metabolism ; Transfection

OBJECTIVETo investigate the effect of losartan on prostatic hyperplasia in spontaneous hypertension rats (SHRs) and its pathophysiological mechanism.

METHODSWe randomly divided 36 male SHRs into three groups of equal number to be treated intragastrically with high-dose losartan (30 mg per kg per d), low-dose losartan (15 mg per kg per d) and distilled water (control group). After 6 weeks of intervention, we measured the body weight and tail artery blood pressure of the rats and compared them with the baseline data. We collected blood from the heart for determination of the levels of serum angiotensin II (Ang II), insulin-like growth factor-1 (IGF-1) and interleukin-6 (IL-6) by enzyme-linked immunosorbent assay (ELISA), and harvested their prostates for measurement of their weight, observation of the tissue ultrastructures under the electron microscope and detection of the expression of endothelial nitric oxide synthase (eNOS) in the prostate tissue by immunohistochemistry.

RESULTSCompared with the control group, the low- and high-dose losartan groups showed significant decreases in systolic blood pressure ([203.75 +/- 10.28] vs [184.54 +/- 16.90] mmHg, P = 0.013; [203.75 +/- 10.28] vs [166.88 +/- 14.74] mmHg, P = 0.001) and diastolic blood pressure ([151.58 +/- 9.96] vs [136.71 +/- 14.28] mmHg, P = 0.022; [151.58 +/- 9.96] vs [122.71 +/- 11.56] mmHg, P < 0.001) of the lower tail artery after treatment, as well as in the prostate weight ([0.73 +/- 0.08] vs [0.64 +/- 0.10] mg, P = 0.011; [0.73 +/- 0.08 ] vs [0.50 +/- 0.17] mg, P < 0.001). Electron microscopy revealed edema of the basal and columnar epithelial cells, concentrated and marginated heterochromatin and widened nuclear gap of interstitial fibroblast nuclei, and reduced mitochondria and endoplasmic reticula in the low-dose losartan group, and even more obvious in the high-dose group. The level of serum Ang II was remarkably higher in the low- and high-dose losartan groups than in the control ([61.32 +/- 2.49] vs [54.85 +/- 7.20] pg/ml, P = 0.021; [65.49 +/- 6.78] vs [54.85 +/- 7.20] pg/ml, P < 0.001]) , that of serum IGF-1 was lower in high-dose losartan than in the control group ([1.50 +/- 0.11] vs [1.60 +/- 0.10] ng/ml, P = 0.03), but the serum IL-6 levels exhibited no significant differences among the three groups. The expression of eNOS in the prostate tissue was significantly higher in the losartan groups than in the controls (P = 0.022), even higher in the high-dose than in the low-dose group.

CONCLUSIONLosartan can suppress the progression of prostate hyperplasia in spontaneous hypertension rats by inhibiting RAS, IGF-1 and angiogenesis.

Angiotensin II ; blood ; Animals ; Antihypertensive Agents ; pharmacology ; therapeutic use ; Hypertension ; drug therapy ; metabolism ; pathology ; Insulin-Like Growth Factor I ; metabolism ; Interleukin-6 ; blood ; Losartan ; pharmacology ; therapeutic use ; Male ; Nitric Oxide Synthase Type III ; metabolism ; Prostate ; drug effects ; metabolism ; pathology ; Prostatic Hyperplasia ; drug therapy ; metabolism ; pathology ; Rats ; Rats, Inbred SHR

BACKGROUNDThe goal of total knee arthroplasty (TKA) is to restore knee kinematics. Knee prosthesis design plays a very important role in successful restoration. Here, kinematics models of normal and prosthetic knees were created and validated using previously published data.

METHODSComputed tomography and magnetic resonance imaging scans of a healthy, anticorrosive female cadaver were used to establish a model of the entire lower limbs, including the femur, tibia, patella, fibula, distal femur cartilage, and medial and lateral menisci, as well as the anterior cruciate, posterior cruciate, medial collateral, and lateral collateral ligaments. The data from the three-dimensional models of the normal knee joint and a posterior-stabilized (PS) knee prosthesis were imported into finite element analysis software to create the final kinematic model of the TKA prosthesis, which was then validated by comparison with a previous study. The displacement of the medial/lateral femur and the internal rotation angle of the tibia were analyzed during 0-135° flexion.

RESULTSBoth the output data trends and the measured values derived from the normal knee's kinematics model were very close to the results reported in a previous in vivo study, suggesting that this model can be used for further analyses. The PS knee prosthesis underwent an abnormal forward displacement compared with the normal knee and has insufficient, or insufficiently aggressive, "rollback" compared with the lateral femur of the normal knee. In addition, a certain degree of reverse rotation occurs during flexion of the PS knee prosthesis.

CONCLUSIONSThere were still several differences between the kinematics of the PS knee prosthesis and a normal knee, suggesting room for improving the design of the PS knee prosthesis. The abnormal kinematics during early flexion shows that the design of the articular surface played a vital role in improving the kinematics of the PS knee prosthesis.

Adult ; Arthroplasty, Replacement, Knee ; methods ; Biomechanical Phenomena ; Female ; Humans ; Knee Prosthesis

OBJECTIVETo observe the expression pattern of albumin during the hepatocyte differentiation by human bone marrow stem cells in vitro.

METHODSHuman bone marrow cells were harvested and cultured in the presence of hepatocyte growth factor (HGF), fibroblast growth factor (FGF) and lymphocyte inhibitory factor (LIF). Cells were stained immunohistochemically by albumin specific antibody and examined under a confocal microscope. Supernatant albumin level was measured biochemically on a serial time points of the culture.

RESULTSBy this condition, the attached cells became mature morphologically in 1 week of culture. Hepatocyte-specific albumin could be detected in mature cells. The albumin level revealed a time-dependent change during a 4-week culture.

CONCLUSIONHuman bone marrow cells could be induced to differentiate to mature hepatocytes that produce and secret albumin in vitro. These cells may contribute to a stable source of hepatocytes for clinical hepatocyte transplantation and artificial liver support system.

Albumins ; biosynthesis ; Bone Marrow Cells ; cytology ; Cell Differentiation ; drug effects ; Cells, Cultured ; Culture Media, Conditioned ; pharmacology ; Fibroblast Growth Factors ; pharmacology ; Hepatocyte Growth Factor ; pharmacology ; Hepatocytes ; cytology ; Humans ; Mesenchymal Stromal Cells ; cytology ; physiology

Special designed group I intron ribozymes can specifically splice objective RNA, repair the mutant gene in RNA level. The specificity of ribozyme is determined by nucleotides specific internal guide sequence (IGS) introduced to the enzyme. In this study, fragment sequence containing Tetrahymena thermophilia intron I of 26S rRNA gene was cloned and cis-splicing activity of this ribozyme was confirmed by in vitro transcription. For evaluating the trans-splicing activity of this ribozyme, a truncated mutant Green Fluorescence Protein (GFP) vector, XYQ5/XYQ10- pEGFP-C2, was constructed. This vector deleted the 3' end 564bp fragment of EGFP coding sequence, led to the lost the activity of emitting green fluorescence. Trans-splicing ribozyme plasmids ptrans-rib-CMV2 for remedy of the truncated mutant EGFP was constructed by PCR and molecular cloning techniques. This vector utilizing cloned 26S rRNA intron 1 as core enzyme; selecting T-G site at 194bp of EGFP coding sequence as splicing receptor, designed an IGS which is inversely complement to the 188-193nt of EGFP mRNA; the 195-890bp fragment of EGFP coding sequence was ligated to the 3'-end of ribozyme core. The fragment containing these components was inserted to a eukayotic expression vector pRC-CMV2. Using linearized XYQ5/XYQ10- pEGFP-C2 and ptrans-rib-CMV2 as templates, truncated EGFP mRNA and the constructed ribozyme vector were transcribed and mixed to evaluate the trans-splicing activity. Analysis of in vitro transcription products mix by RT-PCR verified the existence of wild type EGFP mRNA molecule. Co-transfection of XYQ5/XYQ10- pEGFP-C2 with ptrans-rib-CMV2 to Hela cells proved this ribozyme restored green fluorescence within cell, but the efficiency was low.
Animals ; Base Sequence ; Green Fluorescent Proteins ; genetics ; HeLa Cells ; Humans ; Introns ; genetics ; Molecular Sequence Data ; Mutant Proteins ; genetics ; Mutation ; RNA, Catalytic ; genetics ; RNA, Messenger ; genetics ; Tetrahymena ; enzymology ; Trans-Splicing ; Transcription, Genetic

OBJECTIVETo investigate the clinicopathological features, surgical treatment and prognosis of primary carcinoma of the duodenum.

METHODSThe clinicopathological data of 86 patients with primary duodenal carcinoma from January 1996 to June 2007 were retrospectively reviewed and analyzed by SPSS 13.0.

RESULTSThe clinical manifestation includes upper abdominal pain, jaundice, anemia, gastrointestinal obstruction, melena and weight loss. Four patients had a tumor located in the first portion of the duodenum, 66 in the second portion, 12 in the third portion and 4 in the fourth portion. The preoperative correct diagnostic rate by BUS was 41.7%, by CT 69.4%, by MRI 75.0%, by duodenal endoscopy 84.0%, and by air barium double radiography 80.9%. Complete resection of the tumors was achieved in 38 patients, palliative resection in 45 cases, and exploration alone in 3 cases. The median survival time of the group with complete resection was 42 months versus 13 months in the group with palliative resection, with a significant difference between the two groups (P < 0.05).

CONCLUSIONPrimary carcinoma of the duodenum has no specific symptoms. Early diagnosis and complete resection are effective to improve prognosis.

Adult ; Aged ; Chemotherapy, Adjuvant ; Duodenal Neoplasms ; diagnosis ; drug therapy ; pathology ; surgery ; Duodenum ; pathology ; surgery ; Female ; Follow-Up Studies ; Humans ; Lymphatic Metastasis ; Male ; Middle Aged ; Palliative Care ; Retrospective Studies ; Survival Rate

OBJECTIVETo observe the viability and function of human bone marrow stem cell-derived hepatocytes following cryopreservation in vitro.

METHODSHuman bone marrow cells were induced to differentiate into hepatocytes in the presence of multiple factors. Mature hepatocytes were cryopreserved in 90% FBS and 10% DMSO (Group A), 10% FBS, 30% glycerol and 60% conditioned medium (Group B), and 10% FBS, 10% DMSO, and 80% UW solution (Group C). The cells were thawed after 4 weeks, and the cell viability and the albumin level were determined.

RESULTSThe human bone marrow derived hepatocytes maintained functional morphology after thawing. The viabilities in Group A, B and C were (60.0 +/- 3.3)%, (91.0 +/- 2.6)%, and (89.0 +/- 1.4)%, respectively. After culturing for 24 h, the albumin levels in Group A, B and C were (0.210 +/- 0.005) g/L, (0.340 +/- 0.020) g/L and (0.330 +/- 0.030) g/L, respectively.

CONCLUSIONSHuman bone marrow stem cell-derived hepatocytes can maintain the viability and function after cryopreservation. These cells may contribute to an important source of hepatocytes for clinical hepatocyte transplantation and artificial liver support system.

Adult ; Bone Marrow Cells ; cytology ; physiology ; Cell Culture Techniques ; Cell Differentiation ; Cell Survival ; drug effects ; Cells, Cultured ; Cryopreservation ; methods ; Cryoprotective Agents ; pharmacology ; Hepatocytes ; cytology ; physiology ; transplantation ; Humans ; Myeloid Progenitor Cells ; cytology ; physiology

OBJECTIVETo assess the efficacy and safety of Saw Palmetto Extract Capsules in the treatment of benign prostatic hyperplasia (BPH).

METHODSWe conducted a multi-centered open clinical study on 165 BPH patients treated with Saw Palmetto Extract Capsules at a dose of 160 mg qd for 12 weeks. At the baseline and after 6 and 12 weeks of medication, we compared the International Prostate Symptom Scores (IPSS), prostate volume, postvoid residual urine volume, urinary flow rate, quality of life scores (QOL), and adverse events between the two groups of patients.

RESULTSCompared with the baseline, both IPSS and QOL were improved after 6 weeks of medication, and at 12 weeks, significant improvement was found in IPSS, QOL, urinary flow rate, and postvoid residual urine. Mild stomachache occurred in 1 case, which necessitated no treatment.

CONCLUSIONSaw Palmetto Extract Capsules were safe and effective for the treatment of BPH.

Capsules ; Humans ; Male ; Plant Extracts ; adverse effects ; therapeutic use ; Prostatic Hyperplasia ; drug therapy ; Quality of Life

OBJECTIVETo study the role of slow-release 5-fluorouracil implantation in treatment of unresectable pancreatic cancer.

METHODS85 cases of untreated patients with locally advanced pancreatic cancer (LAPC) were randomized into two groups: Trial group: slow-release 5-fluorouracil implantation (50 patients) and control group (35 patients). Observing the objective tumor response, clinical benefit response, toxicity, complications and survival of patients of the two groups.

RESULTSIn the trial group the overall response rate (PR + NC) was 76.0%, and the clinical benefit response rate was 52.0%. No toxicity was observed. Pancreatic fistula occurred in 2 patients. The median survival time of the two groups was 9.0 months and 4.0 months, respectively. The survival rates of 6- and 12-month were 56.8% vs. 31.4% and 22.9% vs. 2.9% in the two groups, respectively (P = 0.012).

CONCLUSIONSlow-release 5-fluorouracil implantation is a simple, safe and effective method in treatment of LAPC.

Antimetabolites, Antineoplastic ; administration & dosage ; therapeutic use ; Drug Implants ; adverse effects ; Female ; Fluorouracil ; administration & dosage ; therapeutic use ; Follow-Up Studies ; Humans ; Male ; Microspheres ; Middle Aged ; Neoplasm Staging ; Pancreatic Fistula ; etiology ; Pancreatic Neoplasms ; drug therapy ; pathology ; Prospective Studies ; Remission Induction ; Survival Rate

Objective:To investigate the rehabilitation human resources in Shanghai. Methods:Based on World Health Orgnization Rehabilitation in Health Service System: Action Guide, Rehabilitation Information Collection Template of Supporting Tools for Systematic Assessment of Rehabilitation Status was implemented by Shanghai Rehabilitation Medical Association in accordance with the requirements of Shanghai Rehabilitation Treatment Quality Control Center to investigate the number and knowledge structure of rehabilitation-related practitioners in Shanghai. Results:Totally, 311 medical institutions were investigated and 254 institutions set up rehabilitation medicine department, accounting for 82.47%. There were 1414 rehabilitation practitioners and 528 registered rehabilitation physicians, accounting for 37.34%; among them, senior, intermediate and junior professional titles were 142 (26.89%), 262 (49.62%) and 124 (23.48%), respectively. There were 2668 rehabilitation therapists, among which the senior, intermediate and junior professional titles were 11 (0.41%), 461 (17.28%) and 2196 (82.31%), respectively. There were 1614 rehabilitation nurses, among which the senior, intermediate and junior professional titles were 13 (0.81%), 476 (29.49%) and 1125 (69.70%), respectively. There were 2.17 rehabilitation physicians, 11.11 rehabilitation therapists and 6.65 nurses per 100 000 population, which is nearly 1/5 of the international level. There were 121 institutions without rehabilitation medical practitioners and 40 multi-point rehabilitation medical practitioners. Conclusion:There is a big gap in rehabilitation physicians, therapists and nurses in Shanghai, compared with the international level. Based on the imperfect occupational series and qualification certification of physiotherapists, occupational therapists, speech therapists and other specialties, it is suggested to conduct targeted job training in multiple channels and forms, especially to strengthen standardized job training and rehabilitation transformation training for rehabilitation practitioners. It is needed to promote the standardized training of resident and specialist rehabilitation practitioners more quickly to meet the needs of clinical rehabilitation work.