Objective: To explore an ideal method for culturing autologous chondrocytes while maintaining their cartilaginous phenotype by combining monolayer culture with three-dimensional culture. Methods: Chondrocytes were cultured by routine monolayer culture method, then the chondrocytes of fourth passage were seeded into alginate beads for three-dimensional culture following standard protocols. The cell morphology was observed by inverted microscope at predefined time points. The cell viability and proliferation were tested by trypan blue and LIVE/DEAD® Kit, respectively. And extracellular matrix formation was examined by toluidine blue staining. Results: The cartilaginous phenotype of chondrocytes could only be maintained by passage 2 and 3 after monolayer culture. Three-dimensional culture could gradually regain and maintain the cartilaginous phenotype of de-differentiated chondrocytes (P4). Cells of P1-3 by monolayer culture could maintain 90% activity, which decreased from P4. The cytoactivity of cells by three-dimensional culture method was always above 90%. The total doubling rate till P6 by monolayer culture (28 days) was 44 times that by three-dimensional culture. The staining intensity of cartilaginous extracellular matrix was significantly decreased after P4 by monolayer culture (P<0.05). For three-dimensional method, the staining intensities of cartilaginous extracellular matrix at day 21 and 28 were significantly higher than that at day 7 (P<0.05). Conclusion: Monolayer culture allows proper proliferation of chondrocytes, and following three-dimensional culture in alginate beads can regain the cartilaginous phenotype of chondrocytes, thus achieving the aim of both amplification and maintenance of the cartilaginous phenotype.

AIM: To assess the changes of macular thickness of acquire immunodeficiency syndrome ( AIDS) patients.
METHODS:The study based on the data analysis of 38 human immunodeficiency virus ( HIV ) positive patients ( 64 eyes ) . According to CD4 count and whether cytomegalovirus retinitis ( CMVR) happened, the patients were divided into 3 groups. Group A included 16 patients (32 eyes), in which CD4 count was < 50cells/μL and CMVR did not happen. Group B included 10 patients ( 20 eyes), in which CD4 count was>50cells/μL and CMVR did not happen. Group C included 12 patients (12 eyes), in which CD4 count was < 50cells/μL and CMVR just happened. Group D included 10 healthy people ( 20 eyes ) , as normal control group. By using optical coherence tomography ( OCT ) , thickness of retina along with a 1mm diameter circle centered on macular was evaluated.
RESULTS:The mean foveal thicknesses in groups A, B, C and D were 254. 03±15. 63μm, 263. 11±17. 12μm, 304. 50±50. 62μm and 257. 64±8. 54μm in order. Compared with foveal thickness in each group, there were significant differences in general (F=12. 933, P=0. 000). The mean foveal thickness in groups CMVR increased, which was of significant difference (P= 0. 000), compared with other groups.
CONCLUSION: CMVR can impair the structure and function of macular, which then seriously affects the visual function of patients. It's helpful to understand the progress and prognosis of CMVR disease by observing macular structure with OCT in early time.

OBJECTIVETo evaluate the therapeutic effect of Longjintonglin Capsules on type IIIA prostatitis accompanied by abnormal semen liquefaction.

METHODSWe selected 140 patients with type IIIA prostatitis accompanied by abnormal semen liquefaction according to the diagnostic standards of the American Institutes of Health (NIH) and treated them with Longjintonglin Capsules orally 3 capsules once tid for 12 weeks. We obtained the NIH Chronic Prostatitis Symptom Indexes (NIH-CPSI), traditional Chinese medicine (TCM) syndrome scores, leukocyte count in the expressed prostatic secretion (EPS), semen liquefaction time, and the results of semen analysis and compared these indicators before and after the treatment.

RESULTSOf the 140 cases, 132 were included in this study, excluding 8 due to their incomplete case histories. Before and after 4, 8 and 12 weeks of medication, the total NIH-CPSI scores were 24.52 ± 5.43, 21.28 ± 4.85, 18.01 ± 4.28, and 14.49 ± 3.65 (P < 0.01), the TCM syndrome scores were 35.63 ± 6.07, 26.66 ± 5.03, 17.37 ± 4.18, and 11.11 ± 3.96 (P < 0.01), and the leukocyte counts (/HP) were 27.50 ± 7.01, 22.38 ± 5.22, 16:76 ± 4.10, and 11.40 ± 4.74 (P < 0.01), respectively. After 12 weeks of treatment, 31 of the patients with type IIIA prostatitis were cured and another 72 well responded, with an overall response rate of 78.0%. Of those with abnormal semen liquefaction, 61 were cured, 39 well responded, and 32 failed to respond, with an overall effectiveness rate of 75.8%. Semen analysis showed significantly increased percentage of progressively motile sperm after 4, 8 and 12 weeks of medication as compared with the baseline (P < 0.01). No abnormal liver or renal function or other adverse reactions were observed during the treatment.

CONCLUSIONLongjintonglin Capsules, with its advantages of safety, effectiveness and no obvious adverse effects, deserve to be recommended for the treatment of type IIIA prostatitis accompanied by abnormal semen liquefaction.

Capsules ; Drugs, Chinese Herbal ; Humans ; Male ; Medicine, Chinese Traditional ; Phytotherapy ; Prostatitis ; classification ; drug therapy ; Semen ; Semen Analysis

Aged ; Angioplasty, Balloon, Coronary ; Coronary Angiography ; Coronary Artery Disease ; diagnosis ; therapy ; Coronary Vessel Anomalies ; diagnosis ; therapy ; Female ; Humans

OBJECTIVETo investigate the clinical characteristics, pathological and biological behaviors of oral squamous cell carcinomas (OSCC) originated from oral submucous fibrosis (OSF).

METHODSA total of 528 patients with OSCC treated at Department of Oral and Maxillofacial Surgery, Xiangya Hospital, Central South University from 2002 to 2010 were divided into two groups, OSCC originated from OSF and OSCC not originated from OSF. The clinical data regarding age, gender, location, pathological type, metastasis and recurrence rate of these patients were analyzed retrospectively.

RESULTSThe mean age of the group of OSCC originated from OSF (45.8 years) was younger than that of the group of OSCC not originated from OSF (55.9 years). The male female ratio of the group of OSCC originated from OSF (32.3:1) was higher than that of the group of OSCC not originated from OSF (2.3:1). The metastasis and recurrence rate of the group of OSCC originated from OSF (13.5%, 39.1%) was higher than that of the group of OSCC not originated from OSF (7.6%, 27.8%.).

CONCLUSIONSOSCC originated from OSF occurs at younger age and more in male, and is clinically more invasive and metastatic.

Adolescent ; Adult ; Age Factors ; Aged ; Carcinoma, Squamous Cell ; pathology ; Cell Transformation, Neoplastic ; pathology ; Female ; Follow-Up Studies ; Humans ; Lymphatic Metastasis ; Male ; Middle Aged ; Mouth Neoplasms ; pathology ; Neoplasm Recurrence, Local ; Neoplasm Staging ; Oral Submucous Fibrosis ; pathology ; Retrospective Studies ; Sex Factors ; Young Adult

BACKGROUNDThe neurogenic bladder dysfunction caused by spinal cord injury is difficult to treat clinically. The aim of this research was to establish an artificial bladder reflex arc in rats through abdominal reflex pathway above the level of spinal cord injury, reinnervate the neurogenic bladder and restore bladder micturition.

METHODSThe outcome was achieved by intradural microanastomosis of the right T13 ventral root to S2 ventral root with autogenous nerve grafting, leaving the right T13 dorsal root intact. Long-term function of the reflex arc was assessed from nerve electrophysiological data and intravesical pressure tests during 8 months postoperation. Horseradish peroxidase (HRP) tracing was performed to observe the effectiveness of the artificial reflex.

RESULTSSingle stimulus (3 mA, 0.3 ms pulses, 20 Hz, 5-second duration) on the right T13 dorsal root resulted in evoked action potentials, raised intravesical pressures and bladder smooth muscle, compound action potential recorded from the right vesical plexus before and after the spinal cord transaction injury between L5 and S4 segmental in 12 Sprague-Dawley rats. There were HRP labelled cells in T13 ventral horn on the experimental side and in the intermediolateral nucleus on both sides of the L6-S4 segments after HRP injection. There was no HRP labelled cell in T13 ventral horn on the control side.

CONCLUSIONUsing the surviving somatic reflex above the level of spinal cord injury to reconstruct the bladder autonomous reflex arc by intradural microanastomosis of ventral root with a segment of autologous nerve grafting is practical in rats and may have clinical applications for humans.

Anastomosis, Surgical ; Animals ; Atropine ; pharmacology ; Male ; Models, Theoretical ; Rats ; Rats, Sprague-Dawley ; Reflex, Abdominal ; drug effects ; physiology ; Trimethaphan ; pharmacology ; Urinary Bladder, Neurogenic ; physiopathology

OBJECTIVETo analyze the prevalence and risk factors of metabolic syndrome (MS) in Xinjiang Uygur adults.

METHODSWith cluster random sampling, investigations including questionnaire, physical examination and blood testing were performed among 3442 Uygur adults among in Kashgar of Xinjiang on November 2010. Prevalence of MS in groups with different characteristics were calculated and non-conditional logistic regression analysis were used to analyze the risk factors.

RESULTSThe prevalence of MS was 21.2% (728/3442), and the age-adjusted prevalence was 18.5%. The prevalence among males and females was 14.5% (245/1694) (age-adjusted prevalence 12.7%) and 27.6% (483/1748) (age-adjusted prevalence 24.4%) respectively (P < 0.05). The prevalence of MS among 18 to 24 years old and 65 years old and above were 4.3% (21/490) and 28.9% (109/377) respectively. The prevalence of MS increased with age (χ(2) = 204.13, P < 0.05). The prevalence of low blood HDL-C, central obesity, hypertension, hypertriglyceridemia and hyperglycemia was 57.5% (1978/3442), 44.5% (1531/3442), 27.5% (948/3442), 20.2% (696/3442) and 8.6% (297/3442) respectively. Compared to age group 18 - 24, the risk of MS occurrence was higher in age group 25 - 34, 35 - 44, 45 - 54, 55 - 64 and 65 years-old above, the according OR (95%CI) values were 2.29 (1.38 - 3.81), 6.91 (4.31 - 11.09), 10.81 (6.72 - 17.40), 12.52 (7.74 - 20.26) and 10.20 (6.20 - 16.78), respectively. Smoking also increased the risk of MS (OR = 2.35, 95%CI: 1.64 - 3.37).

CONCLUSIONThe prevalence of MS in Xinjiang Uygur was in high level; The prevalence of MS is higher in female than in male; The risk factors of MS included female, age and smoking.

Adolescent ; Adult ; Aged ; China ; epidemiology ; Female ; Humans ; Male ; Metabolic Syndrome ; ethnology ; Middle Aged ; Minority Groups ; Prevalence ; Risk Factors ; Young Adult

Animals ; Cytoskeletal Proteins ; biosynthesis ; genetics ; Drugs, Chinese Herbal ; pharmacology ; Hepatocytes ; metabolism ; ultrastructure ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo observe the influence of high blood glucose fluctuation on the endothelial function of type 2 diabetes mellitus (T2DM) rats and the effects of Panax Quinquefolius Saponin (PQS) of stem and leaf.

METHODSThe T2DM model was induced by intraperitoneal injection of a small dose of streptozotocin (STZ, 35 mg/kg) plus high fat and high caloric laboratory chow. Then, diabetic rats were divided into steady high blood glucose (SHG) group and fluctuant high blood glucose (FHG) group according to fasting blood glucose coefficient of variation (FBG-CV), and then, the FHG group rats were divided into 4 groups according to the level of FBG-CV and fasting blood glucose: PQS 30 mg/(kg·d) group, PQS 60 mg/(kg·d) group, metformin hydrochloride control (MHC) group, and FHG control group, 10 in each group. Meanwhile, 10 rats without any treatment were used as normal control (NOR) group. Eight weeks later, the aortic arteries histology, plasma hepatocyte growth factor (HGF), and serum nitric oxide (NO), endothelin-1 (ET-1), tumor necrosis factor α (TNF-α), and soluble intercellular adhesion molecule 1 (sICAM-1) were measured.

RESULTSIn comparison with the NOR group, the level of plasma HGF and serum NO, ET-1 and TNF-α, and sICAM-1 in SHG and FHG control groups were all significantly increased (P<0.01); in comparison with the SHG group, plasma HGF and serum NO, ET-1, TNF-α, and sICAM-1 in FHG group were all significantly increased further (P<0.01 or P<0.05); meanwhile, in comparison with the FHG control group, the level of plasma HGF and serum NO, ET-1, TNF-α, and sICAM-1 in PQS and MHC groups were all decreased significantly (P<0.01). However, comparison of the aortic arteries histology among groups showed no significant differences either before or after treatment.

CONCLUSIONBlood glucose fluctuation could facilitate the development of vascular endothelial dysfunction in T2DM rats, while PQS could improve the endothelial function of T2DM rats with high blood glucose fluctuation, which may be related to its effects of relieving vessel stress, decreasing vasoconstrictor ET-1 production, preventing compensated increase of NO, and reducing inflammatory reaction.

Animals ; Aorta ; drug effects ; pathology ; Blood Glucose ; metabolism ; Body Weight ; drug effects ; Diabetes Mellitus, Type 2 ; blood ; drug therapy ; Endothelin-1 ; blood ; Endothelium, Vascular ; drug effects ; physiopathology ; Hepatocyte Growth Factor ; blood ; Intercellular Adhesion Molecule-1 ; blood ; Male ; Nitric Oxide ; blood ; Panax ; chemistry ; Plant Leaves ; chemistry ; Plant Stems ; chemistry ; Rats ; Saponins ; pharmacology ; therapeutic use ; Solubility ; Tumor Necrosis Factor-alpha ; blood

OBJECTIVETo explore the effects of Panax Quinquefolium Saponin (PQS) on phosphatidylinositol 3-kinase/serine threonine kinase (PI3K/Akt) pathway of neonatal rat myocardial cells subjected to hypoxia.

METHODSNeonatal rat myocardial cells were cultured in vitro. After the myocardial cell injury was induced by hypoxia, the cells were randomized into 5 groups: the normal group, the model group, the positive control group (Ciclosporin A, 2 µ mol/L), the low-dose PQS group (PQSL, 25mg/L), and the high-dose PQS group (PQSH, 50 mg/L). Morphology and behavior of myocardial cells were observed under an inverted microscope. Apoptosis rate and lactate dehydrogenase (LDH) leakage rate of myocardial cells were determined by colorimetry. Mitochondrial transmembrane potential was assessed using a fluorexon laser. Phospho-glycogen synthase kinase (GSK)-3β and phospho-Akt as well as cytochrome C were determined by Western blot

RESULTSLDH leakage in the Ciclosporin A group, PQSH group and PQSL group reduced progressively compared with the model group (P<0.05). Akt and GSK-3β was strongly phosphorylated after treatment with Ciclosporin A and PQS compared with the model group (P<0.05, P<0.01). Compared with the model group (16.41±1.74; 35.28±6.30), both the integrated optical density of mitochondrial permeability transition pore (MPTP) and the mitochondrial transmembrane potential significantly increased in the PQSH group (42.74±2.12; 71.36±6.54) and the PQSL group (39.58±1.49; 66.99±5.45; P<0.05, P<0.01). However, the protein of cytochrome C outside the mitochondrion decreased in the PQSH group (273.66±14.61) and the PQSL group (259.62±17.31) compared with the model group (502.41±17.76; P<0.05).

CONCLUSIONThrough activation of the PI3K/Akt pathway and inhibition of the MPTP, PQS might protect the heart against ischemia injury and apoptosis of myocardial cells.

Animals ; Animals, Newborn ; Cell Hypoxia ; drug effects ; Cell Shape ; drug effects ; Cell Survival ; drug effects ; Cells, Cultured ; Glycogen Synthase Kinase 3 ; metabolism ; Glycogen Synthase Kinase 3 beta ; L-Lactate Dehydrogenase ; metabolism ; Membrane Potential, Mitochondrial ; drug effects ; Mitochondria ; drug effects ; metabolism ; Mitochondrial Membrane Transport Proteins ; metabolism ; Myocytes, Cardiac ; cytology ; drug effects ; enzymology ; Phosphatidylinositol 3-Kinases ; metabolism ; Phosphorylation ; drug effects ; Protein-Serine-Threonine Kinases ; metabolism ; Rats, Sprague-Dawley ; Saponins ; pharmacology ; Signal Transduction ; drug effects