Objective:To study the isolation, culture and identification of the TMJ cells and to observe the biological characteristics of cultured fibrochondrocytes. Methods:The TMJ discs were dissected from two 1 month goats under sterile conditions and were digested with collagenase. The cells were collected. Morphological changes and attachment efficiency were constantly observed under phase-contrast microscope. Immunohistochemical staining for type I collagen as well as toluidine blue staining were performed. Ultrastructures of the TMJ cells were observed under transmission electron microscope. Results: Most of the primary fibrochondrocytes presented a short spindle-shape while the rest showed polygon-shape. On the 7th day, the perliferating fibrochondrocytes started to contact each other to form a monolayer covering the bottom of the incubation disc. Immunohistochemical staining of type I and toluidine blue staining exhibited positive results. The fibrochondrocytes cytoplasms were rich in mictochondria and endoplasm reticulum. Conclusion: The fibrochondrcytes isolated from one-month-old goat TMJ disc have good proliferation ability in vitro and cells from passage 1 to 3 might be used as seed cells for TMJ disc tissue engineering.

BACKGROUND:There is no common cognition in the cell type in the temporomandibular joint(TMJ)discs,and names describing TMJ disc cells also vary a lot.OBJECTIVE:To characterize the type and the distribution of cells in the TMJ disc of goats DESIGN,TIME AND SETTING:A single sample observation was completed in Cettutar and Motecutar Biologicat Center and Electron Microscope Center of Lanzhou University from March to May in 2007.MATERlALS:TMJ discs were obtained from two one-month-old healthy goats that were slaughtered freshly.METHODS:Bilateral TMJ discs of goats were cut off completely and were divided into 6 parts by 3 cuts in the major axis direction (mediolaterally)and 2 cuts in the minor axis direction(anteroposteriorly).Then the marked samples were fixed in 10%neutral formalin Iiquid for 24 hours and embedded by paraffin.MAIN OUTCOME MEASURES:Hematoxylin and eosin staining were used to identify regional variation of cell type and cellnumbers.Toluidine blue staining and collagen type Ⅰimmunohistochemical assay were performed to test the distribution of collagens.Transmission etectren microscopy was used to observe the ultrastructure of cells of goat TMJ discs.RESULTS:TMJ discs were comprised of cells and collagen fibers distributing unevenly.Collagens were mostly type Ⅰ.Collagen fibers were wave or crimping and approximately parallel to each other.with cells scattered in their matrix.Fibroblast-like cells and chondrocyte-like cells were the main two types of cells existing,with the former predominating over the later in a ratio of 2.05:1 approximately.There were no significant regional differences in cell type and distribution statistically.Transmission electron microscopy denoted that fibroblast-iike cells have fairly larger fusiform or irregular nuclei with very few organelles,while the chondrocyte-like cells exhibited round or elliptical nuclei,well defined pericellular electron lucent zones,unconspicuous cytocysta and non-distinctive pseudopodia CONCLUSION:There are no significant differences in type,number and arrangement of cells in TMJ discs of one-month-old goats statistically,with Fibroblast-like cells predominating slightly over chondrocyte-like cells.

Objective Inflammation and calcification are common phenomenons in human atherosclerotic plaque.The objective of this study was to determine the gene expression of some proteins and anti-inflammatory factors as well as bone formation mediators in the fibrous cap and shoulder region of non-calcified and calcified carotid endarterectomy(CEA) plaques.Methods Twenty-two CEA plaques were classified as Va(non-calcified,n=11) and Vb(calcified,n=11) in accordance with the AHA consensus in 1995.Mean percent in carotid stenosis and calcification area was 76.4% and 0% in Va and 74.2% and 33.6% in Vb respectively as determined by quantitative histomorphometry.Using laser capture microdissection,the fibrous cap and shoulder regions were excised from 22 frozen sections/plaques.After total RNA extraction and reverse transcription,gene expression of proteins(IL-1,IL-8,and MCP-1),anti-inflammatory factor(IL-10) and bone formation protein(BMP-6 and Osteocalcin) were detected by Real-time PCR.Results mRNA expression of MCP-1,IL-8 and IL-1 in Va plaques were 1.8 fold higher than that in Vb plaques,mRNA expression of anti-inflammatory factor IL-10 and osteogenic related genes BMP-6 and Osteocalcin were 3.8,4.2,and 6.9 fold higher in Vb plaques compared to Va plaques.Conclusions Our findings indicate that fibrous cap inflammation and susceptibility to disruption is more likely to occur in non-calcified plaques.

OBJECTIVETo investigate the clinical value of Paroxetine combined with mid-frequency electrical pulse acupoint stimulation (EPAS) in the treatment of premature ejaculation (PE).

METHODSTotally 69 PE patients were equally assigned to receive oral Paroxetine 20 mg/d, mid-frequency EPAS, or oral Paroxetine 10 mg/d combined with mid-frequency EPAS (P + EPAS) , all for 8 weeks. We obtained the intravaginal ejaculation latency time (IELT) and Chinese Index of Premature Ejaculation (CIPE-5) scores of the patients before and after treatment, and compared adverse reactions among the three groups of patients.

RESULTSOne patient of the Paroxetine group gave up treatment because of abdominal pain and nausea. Compared with the baseline, the patients in the Paroxetine, EPAS, and P + EPAS groups all showed markedly increased IELT ([0.92 ± 0.11] vs [4.07 ± 0.11] min, P < 0.01; [0.92 ± 0.12] VS [2.78 ± 0.17] min P < 0.05; [0.91 ± 0.09] vs [5.31 ± 0.13], P < 0.01) and decreased CIPE-5 scores (12.5 ± 3.0 vs 22.0 ± 2.1, P < 0.01; 12.8 ± 2.9 vs 19.5 ± 1.9, P > 0.05; 13.1 ± 2.8 vs 25.2 ± 2.1, P 0.01), with statistically significant differences between the P + EPAS group and the other two (P < 0.05). The total effectiveness rate was 95.7% in the P + EPAS group, remarkably higher than in the Paroxetine (72.7%, P < 0.05) and the EPAS group (47.8, P < 0.01).

CONCLUSIONOral Paroxetine combined with mid-frequency EPAS has a higher safety and efficacy than either Paroxetine or EPAS alone in the treatment of PE.

Acupuncture Points ; Aged ; Combined Modality Therapy ; methods ; Ejaculation ; Electroacupuncture ; methods ; Humans ; Male ; Paroxetine ; therapeutic use ; Premature Ejaculation ; therapy ; Serotonin Uptake Inhibitors ; therapeutic use ; Treatment Outcome

Objective To evaluate the clinical efficacy of Yiqi Fumai and Zixin Yangyin Prescription combined with Captopril for the treatment of children's hand-foot-mouth disease (HFMD) complicated with viral myocarditis (VMC); To explore the mechanism. Methods Totally 80 cases children with HFMD complicated with VMC were selected and randomly divided into two groups, with 40 cases in each group. Both groups were given conventional low flow oxygen, cardiac function monitoring, ribavirin antiviral, and fructose nutrition, and severe cases were given immune globulin and dexamethasone to relieve symptoms. The control group was given Captopril 0.5 mg/(kg?d), three times a day,orally,on the basis of conventional treatment,and the treatment group was given Yiqi Fumai and Zixin Yangyin Prescription on the basis of control group,one dose a day,decoction 180 mL,6 to 10 times a day,orally. The treatment in both groups lasted for two weeks. Serum hs-CRP, CK, CK-MB and cardiac function change before and after treatment of the two groups were observed. Electrocardiogram curative effect and comprehensive curative effect were evaluated. Results Compared with before treatment, the contents of hs-CRP, CK-MB, and CK in both groups decreased after treatment (P<0.05), and the treatment group was lower than the control group (P<0.05);Compared with before treatment, EF and FS in both groups increased after treatment, and A/E of mitral valve diastolic blood flow velocity peak decreased after treatment, with statistical significance among EF, FS and A/E between the two groups (P<0.05). The total effective rate of electrocardiogram was 92.5% (37/40) in the treatment group and 75.0% (30/40) in the control group, with statistical significance (Z=-2.370, P=0.018). The total effective rate of comprehensive efficacy was 92.5% (37/40) in the treatment group, and 77.5% (31/40) in the control group, with statistical significance (Z=-2.397, P=0.017). Conclusion Yiqi Fumai and Zixin Yangyin Prescription combined with Captopril can improve the clinical symptoms of HFMD complicated with VMC, improve clinical efficacy, whose mechanism may be related to the decrease of contents of hs-CRP, CK and CK-MB and enhancement of cardiac function.

AIMTo investigate the chemosensitivity to lidamycin (C-1027) in mdr1 gene overexpressing cancer cell lines established by drug induction and by gene-transfection.

METHODSDNA was cloned by RT-PCR and then eukaryotic expressing recombinant plasmid pcDNA3. 1/mdrl was constructed. Using Lipofectamine 2000, a strain of stably transfected human hepatoma cancer cells, HepG2/mdrl, was obtained. The mdr1 mRNA level, P-glycoprotein (P-gp) level and the activity of P-gp to extrude drugs in cancer cells were determined by RT-PCR, immunofluorescence analysis and rhodamine 123 efflux assay. The chemosensitivity of cancer cells with low or high mdr1 expression to lidamycin and other antitumor drugs was tested by MTT assay.

RESULTSThe mdr1 mRNA and P-gp levels in KBv200, MCF-7/ADR, and stably transfected HepG2/mdr1 cells were much higher than that in respective parent KB, MCF-7 and HepG2 cells. The IC50 values of lidamycin for KBv200, MCF-7/ADR and HepG2/mdrl cells were (0.24 +/- 0.20) nmol x L(-1), (0.028 +/- 0.011) nmol x L(-1), and (0.020 +/- 0.011) nmol x L(-1), respectively. Compared with parental cells, the values of resistant fold for KBv200, MCF-7/ADR and HepG2/mdr1 cells to lidamycin were 6.8, 1.6 and 1.3 fold; to adriamycin were 37.2, 181.3 and 8.8 fold; to taxol were 336.8, 49.2 and 40.3 fold, respectively.

CONCLUSIONLidamycin is highly active to multidrug resistant cancer cells. The chemosensitivity of those resistant cancer cells to lidamycin is approximately at the similar level as that of parent cancer cells.

ATP-Binding Cassette, Sub-Family B, Member 1 ; analysis ; genetics ; Aminoglycosides ; pharmacology ; Antibiotics, Antineoplastic ; pharmacology ; Cell Line, Tumor ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; Enediynes ; pharmacology ; Genes, MDR ; Humans ; Neoplasms ; drug therapy ; pathology ; Transfection

OBJECTIVETo review the mechanisms and current clinical application of pharmacological interventions for phantom limb pain.

DATA SOURCESBoth Chinese and English language literatures were searched using MEDLINE (1982 - 2011), Pubmed (1982 - 2011) and the Index of Chinese Language Literature (1982 - 2011).

STUDY SELECTIONData from published articles about pharmacological management of phantom limb pain in recent domestic and foreign literature were selected. Data extraction Data were mainly extracted from 96 articles which are listed in the reference section of this review.

RESULTSBy reviewing the mechanisms and current clinical application of pharmacological interventions for phantom limb pain, including anticonvulsants, antidepressants, local anaesthetics, N-methyl-D-aspartate receptor antagonists, non-steroidal anti-inflammatory drugs, tramadol, opioids, calcitonin, capsaicin, beta-adrenergic blockers, clonidine, muscle relaxants, and emerging drugs, we examined the efficacy and safety of these medications, outlined the limitations and future directions.

CONCLUSIONSAlthough there is lack of evidence-based consensus guidelines for the pharmacological management of phantom limb pain, we recommend tricyclic antidepressants, gabapentin, tramadol, opioids, local anaesthetics and N-methyl-D-aspartate receptor antagonists as the rational options for the treatment of phantom limb pain.

Analgesics ; therapeutic use ; Analgesics, Opioid ; therapeutic use ; Anticonvulsants ; therapeutic use ; Antidepressive Agents ; therapeutic use ; Humans ; Phantom Limb ; drug therapy ; Tramadol ; therapeutic use

Objective To report our experience with retroperitoneoscopic nephroureterectomy with excision of a bladder-cuff for renal pelvic and ureteral tumors.Methods Thirty-five patients (21 women and 14 men;mean age,67 years;age range,49 -82 years) with upper urinary tract tumors underwent retro- peritoneal laparoscopic nephrourcterectomy with excision of a bladder-cuff.Of the 35 cases,15 had pelvic tumors and 20 had ureteral tumors;19 cases had the tumors on the right side and 16 on the left.Two cases had ureteral tumors combined with bladder tumors.One case had bilateral ureteral tumors then concomitantly had bladder tumors.The needle electrode was used to circleround incise the bladder thoroughly 0.5 cm away from the ureterostoma.Three trocars in the waist were used for dissecting the kidney;and the ureter was dis- sected as far distally downward.Then an incision of 5-9 cm was created in the lower abdomen to allow dis- section of the distal ureter and bladder-cuff and intact specimen extraction.Results The operation was successful in all 35 patients.The mean operative time was 3.1 h ( range,1.5-6.0h).The mean estimated blood loss was 166 ml (range,20-1600 ml).Four cases received blood transfusion.The patient's activity re- covered in 20-32 h after operation.Postoperative pathology showed transitional cell carcinoma in 30 cases, poorly differentiated adenocareinoma in 2 (ureter),squamous cell carcinoma in 1 (ureter),leiomyosarcoma in 1 (ureter),xanthogranulomatous pyelonephritis in 1.Duodenal leakage occurred in 1 patient who had had dialysis a drainage catheter was placed on the third d after operation,and the patient died of heart failure af- ter 2 months.Postoperative vesical irrigation was performed to prevent tumor recurrence.The mean hospital stay was 11 d.During a mean follow-up of 14 months(range,1-32 months),1 patient developed pelvic me- tastasis and was alive with the tumor.The other 33 patients survived free of tumor to date.No patient had re- current transitional cell carcinoma of the bladder.Conclusions Our data demonstrate that retroperitoneo- scopic nephroureterectomy for renal pelvic and ureteral tumors has shorter incision and more rapid postopera- tive recovery compared with open surgery.Using resectoscope to resect the termination of ureter allows more complete excision of the ureter.

·AIM: To observe the therapeutic effect of visual perception training combined with total nutrition meal sequential therapy in the treatment of myopic amblyopia.·METHODS:Totally 73 children ( 135 eyes ) with myopic amblyopia were divided into control group ( 36 cases, 67 eyes) and treatment group (37 cases, 68 eyes) according to random number table. The control group were treated with traditional spectaculars and grating covering combined with fine eyesight training;the treatment group were treated with visual perception training combined with total nutrient meal sequential therapy. The visual acuity, diopter and average diopter of two groups were compared before and after treatment at 3, 6mo and 1a. The curative effect of two groups of children was compared after 1a treatment. And the adverse reactions were recorded in two groups during the treatment period. The recurrence rate of amblyopia in 1a follow-up was compared between two groups.·RESULTS: The difference of visual acuity between two groups was not significant at 3mo (P>0. 05). The visual acuity of the treatment group was significantly higher than that of the control group at 6mo and 1a (P<0. 05). There was no significant difference in diopter between the two groups after 3, 6mo and 1a (P>0. 05), but the average annual refractive changes in the treatment group were significantly lower than that in the control group ( P<0. 05). The basic cure rate and total effective rate of the treatment group were significantly higher than that of the control group (P<0. 05). There were no severe adverse reactions occurred between two groups during the treatment period. The recurrence rate of amblyopia in the treatment group was significantly lower than that in the control group (P<0. 05) after 1a follow-up.· CONCLUSION: Visual perception training combined with total nutrition meal sequential therapy in the treatment of myopic amblyopia in preschool children can significantly improve patients' visual acuity, reduce the average annual diopter changes, improve the therapeutic effect, reduce the recurrence rate of amblyopia.

OBJECTIVETo study the bone marrow microvessel density (MVD) and vascular endothelial growth factor (VEGF) expression and their clinical significance in patients with aplastic anemia (AA).

METHODSBone marrow biopsies in 51 newly diagnosed patients with AA were evaluated the MVD and VEGF expression by immunostaining with anti-factor VIII related antigen and VEGF monoclonal antibodies at regular time points after immunosuppressive therapy (IT).

RESULTSThe mean bone marrow MVD in AA group was 5.5 +/- 3.5, being significantly lower than that in normal control group (8.7 +/- 3.4, P < 0.05). MVDs of SAA and NSAA patients were 7.4 +/- 2.9 and 4.3 +/- 3.4, respectively, being significantly different (P < 0.01). The VEGF expression in AA group was significantly lower than that in control group [(6.7 +/- 8.4)% vs (14.7 +/- 6.1)%, P < 0.01], but there was no difference between SAA and NSAA. Bone marrow MVD and VEGF were significantly increased after IT in 22 responded AA patients.

CONCLUSIONBone marrow MVD and VEGF expression are low in AA patients which may be one of pathophysiologic mechanisms of bone marrow failure in AA. Proangiogenic and ameliorating microcirculation agents together with IT might accelerate the recovery of hematopoiesis in AA patients.

Adolescent ; Adult ; Anemia, Aplastic ; metabolism ; pathology ; Bone Marrow ; blood supply ; metabolism ; Child ; Female ; Humans ; Male ; Microvessels ; pathology ; Middle Aged ; Neovascularization, Pathologic ; Vascular Endothelial Growth Factor A ; metabolism